OFP is a class of plant-specific transcription factors that play important roles in the regulation of plant organ morphogenesis and response to abiotic stresses.In order to study the characterization of soybean OFP transcription factor family members and their roles in drought stress and salt stress,bioinformatics methods were applied to identify and analyze soybean OFP family members.The results showed that:a total of 41 GmOFPs,named GmOFP-1—GmOFP-41,were identified in soybean;these genes were unevenly distributed on 19 chromosomes of soybean,encoding 152—414 amino acids;subcellular localization predicted that soybean OFP proteins were mainly localized in nucleus,chloroplasts,and mitochondria;a total of 10 conserved motifs were identified in soybean OFP proteins,conservative Motifs 1 and 2 were present in all OFP members.Phylogenetic analysis classified soybean and Arabidopsis OFP proteins into five subfamilies ClassⅠ—Class V,of which soybean OFP family genes were mainly distributed in ClassⅠ and Class Ⅲ.The collinearity analysis revealed that 75 pairs of genes in the soybean genome had collinearity,four pairs of genes had tandem duplications,and only two genes,GmOFP-2 and GmOFP-39,did not have collinearity,which indicated that gene fragment duplication was the main reason for the increase in the number of soybean OFP family members.The expression patterns of GmOFP gene family members under drought stress and salt stress treatments were analyzed by qRT-PCR,and the results showed that,compared with the control,16 members out of 41 GmOFP genes exhibited significant differences in gene expression levels after drought treatments,with significant up-regulation of the expression of GmOFP-15,GmOFP-17,and GmOFP-32,while the GmOFP-4,GmOFP-5,GmOFP-6,GmOFP-9,GmOFP-12,GmOFP-21,GmOFP-23,GmOFP-25,GmOFP-26,GmOFP-27,GmOFP-38,GmOFP-39,and GmOFP-40 were significantly down-regulated after drought treatment.Eight members of GmOFPs showed significant differences in gene expression levels after salt treatment,among which GmOFP-7,GmOFP-14,GmOFP-31,GmOFP-32,GmOFP-36,and GmOFP-40 were significantly up-regulated,and GmOFP-1 and GmOFP-15 were significantly down-regulated.The above results suggest that the soybean OFP gene family may have important functions in response to drought stress and salt stress.

To explore the relationship between soil bacterial community diversity and soil enzyme activity of Qinqiong(Q) and Magnum(M) under salinity stress treatment,the bacterial community structure and soil enzyme activity of Qinqiong and Magnum for 0,6,12,24,48 h were compared and analyzed by using high-throughput sequencing of bacterial 16S rRNA and colorimetry.The results indicated that at the phylum level,the relative abundances of Actinobacteria and Proteobacteria in the rhizosphere soil of Qinqiong were higher than those in Magnum.At the genus level,the relative abundances of Bacillus and Arthrobacter in the rhizosphere soil of Qinqiong were greater than those in Magnum,while the relative abundance of Pseudomonas was lower compared to Magnum.After 48 hours of saline-alkali stress,the activities of soil polyphenol oxidase(S-PPO),soil catalase(S-CAT),soil dehydrogenase(S-DHA),and soil invertase(S-AI) in the rhizosphere of Qinqiong were 1.32,1.53,1.38,and 1.28 times higher than those of Magnum,respectively.Furthermore,the S-CAT activity exhibited a significant positive correlation with the bacterial ACE and Chao diversity indices.Qinqiong may influence changes in bacterial community structure by altering the relative abundances of dominant bacterial groups,thereby enhancing the bacterial community diversity index and soil enzyme activity.In contrast,the changes in the bacterial community within the rhizosphere of Magnum were not significant.The diversity of soil microbial communities is a critical factor determining the saline-alkali tolerance of oat varieties.

Salt stress causes a significant threat to crop yield and quality.As one of the pioneer crop species in salt tolerance research,barley holds critical significance;the exploration of its salt tolerance mechanisms is capable of providing a theoretical foundation for crop salt-tolerance breeding programs.Two naked barley landraces,namely B87 with salt-sensitivity and B94 with salt-tolerance,were employed as experimental materials.At the three-leaf stage,their seedlings were exposed to a 200 mmol/L NaCl treatment for 7 days.Subsequent to the treatment,the above-ground tissues were collected for transcriptomic and metabolomic sequencing.By means of integrated multi-omics analysis,this study was designed to elucidate the molecular mechanisms governing salt tolerance in naked barley.The results demonstrated that 2 240 differentially expressed genes (DEGs) and 198 differentially abundant metabolites (DAMs) were identified in B87 via transcriptomic and metabolomic profiling,whereas 923 DEGs and 232 DAMs were detected in B94.Venn diagram analysis further revealed that the salt-tolerant naked barley B94 contained 480 specific DEGs and 129 specific DAMs.Furthermore, GO and KEGG analyses were separately performed on the DEGs and DAMs. And the DEGs of B94 were significantly enriched in 11 unique pathways, while its DAMs were only significantly enriched in 1 unique pathway. In addition, correlation analysis between the transcriptome and metabolome was conducted, and it was found that the changes in genes and metabolites exhibited both consistency and inconsistency. These research efforts not only enhance the current understanding of the molecular mechanisms underlying salt tolerance in naked barley,but also provide valuable insights and candidate targets for the development of salt-tolerant naked barley cultivars in future breeding.

To elucidate the mechanisms of cold response and explore superior cold-tolerance gene resources in wheat,this study employed transcriptome sequencing to uncover key regulatory networks underlying wheat cold response,and performed functional validation of the candidate gene TaGGCT18-6A.The results demonstrated that 4 ℃ cold treatments induced 10 893 and 18 784 differentially expressed genes(DEGs)in wheat seedlings after 6 h and 24 h cold treatment,respectively.KEGG analysis revealed significant enrichment of DEGs in pathways including MAPK signaling transduction and glutathione metabolism.A γ-glutamyl cyclotransferase gene TaGGCT18-6A was cloned through screening key genes in glutathione metabolism.This gene had a length of 1 772 bp,encoding 218 amino acids with conserved GGCT-like superfamily and ChaC core domain.Promoter cis-acting element analysis identified stress-responsive elements such as low-temperature-responsive(LTR)and dehydration-responsive (DRE)elements; consistently,expression pattern analysis showed sustained upregulation of TaGGCT18-6A under 4 ℃ cold treatments. Functional validation in transgenic rice revealed that overexpression lines OE#1, OE#2 and OE#3 exhibited significantly enhanced survival rate,plant height,and biomass. Overexpression lines OE#1 and OE#2 exhibited significantly enhanced plant height, while overexpression lines OE#1, OE#2 and OE#3 exhibited significantly reduced relative electrolyte leakage under -4 ℃ freezing treatments.After 4 ℃ treatment,overexpression lines accumulated higher levels of osmolytes(proline and soluble sugars),decreased malondialdehyde(MDA)content,and increased activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT).These findings collectively demonstrated that TaGGCT18-6A enhanced plant cold tolerance by regulating glutathione metabolism to improve antioxidant capacity.This study will provide a theoretical foundation and valuable genetic resources for molecular breeding of cold-tolerant wheat.

Plant sodium-hydrogen antiporter(NHX,Na⁺/H⁺ antiporter)plays a crucial role in plant sodium and potassium ion balance and cellular pH regulation.In order to investigate the relationship between salt tolerance and ScNHXs,it was conducted to identify and analyze the ScNHXs by bioinformatics process,and to examine the expression pattern of ScNHXs under salt stress by RT-qPCR,which can provide the reference information for the investigation of the potential functions of ScNHXs as well as the mining of salt tolerance genes in rye.A total of 10 rye NHX gene family members(ScNHX1—ScNHX10)were identified,and the phylogenetic tree analysis showed that they could be divided into two subfamilies,Vac and Endo,containing four and six genes,respectively.Physicochemical property analysis of the encoded proteins showed that most of the molecular weight ranged from 27.92 to 59.72 ku,the number of amino acids from 253 to 546 aa,and the isoelectric point between 5.17 and 8.81,with most of proteins being classified as acidic proteins.Signal peptide prediction indicated the absence of signal peptides in the members,and transmembrane structure analysis revealed that all members possessed transmembrane structures.The subcellular localization prediction indicated that ScNHXs were located in the plasma membrane and vesicles.Spatial structure prediction showed that their secondary structures mainly consisted of α-helices and irregular convolutions.Gene structure and motif analyses revealed that the number of exons of the ScNHXs varied from 13 to 24,and all of them possessed a conserved Na⁺/H⁺ exchange structural domain.In addition,cis-acting element analysis revealed that numerous elements related to hormone response and abiotic stresses were found in the promoter region of ScNHXs.Analysis of rye transcriptome data revealed significant differences in the expression patterns of ScNHXs in different tissues of rye.RT-qPCR analysis showed that ScNHXs responded differently to different concentrations of NaCl stress,and were able to persistently respond to salt stress over a long period of time.In summary,ScNHXs may be involved in the biological regulation during salt stress in rye.

Chalcone synthase(CHS)is the initial and crucial enzyme in the flavonoid biosynthesis pathway,responsible for the synthesizing of metabolites such as flavones,flavonols,isoflavones,and anthocyanins,which play a vital role in enhancing plant stress resistance.In order to explore the role of CHS genes in the drought stress response of oat seedlings,it identified a CHS gene from the full-length transcriptome data of oats,named AsCHS.Gene cloning,bioinformatics analysis,subcellular localization,and expression pattern analysis were conducted.The results showed that the AsCHS gene encoded a protein composed of 398 amino acids and had a CHS family-specific tag sequence.This protein was hydrophobic and unstable.It was a non-transmembrane protein and was located in the nucleus and cytoplasm.Secondary structure prediction showed that AsCHS was mainly composed of α-helices and random coils.The analysis of the cis-acting elements within the promoter region revealed that the gene contained cis-elements associated with drought stress response and multiple hormone signaling pathways.Phylogenetic tree analysis showed that AsCHS was closely related to its counterparts in Lolium perenne,Poa annua,and Deschampsia antarctica.Subcellular localization indicated that the AsCHS protein was localized in the nucleus and cytoplasm.Compared with the control group,the expression pattern of AsCHS in oat seedlings under drought stress changed from fluctuating expression to incremental expression with different germination time,shifting from the highest expression level in roots to the highest in leaves,with significant differences observed in leaves expression.It laid a foundation for elucidating the function of AsCHS in the drought stress response of oats.

To investigate the effects of different dosages of biochemical fulvic acid (BFA) on the improvement of soda saline-alkali soil and the response mechanism of maize growth,a pot experiment was conducted using soda saline-alkali soil from Inner Mongolia as the test soil and maize Dongdan 181 as the test variety.Four BFA application rates were set as 0(CK),2(FA2),4(FA4),8 g/kg(FA8).Soil nutrients,microbial diversity,maize salt tolerance,biomass,and other indicators were measured.The results showed that compared to the CK,soil pH decreased with increasing BFA dosage.The soil available phosphorus content increased significantly after the application of BFA,but there was no significant difference among the three treatments of FA2,FA4 and FA8 at 30,62 and 80 days after sowing.Soil salinity increased with the increase of BFA dosage,with an increase of 23.30%—89.32%.Soil exchangeable potassium content increased with increasing BFA dosage,while exchangeable calcium content gradually decreased.The proportion of <0.053 mm silt and clay fractions in the soil decreased by 6.49,9.92 and 13.97 percentage points under FA2,FA4,and FA8 treatments,respectively,compared to the CK treatment.Meanwhile,the proportion of 0.053—0.250 mm aggregates increased by 5.90,8.99 and 13.75 percentage points,the proportion of 0.250—2.000 mm aggregates increased by 0.55,0.87 and 0.21 percentage points,while the proportion of >2.000 mm aggregates increased by 0.04,0.06 and 0.01 percentage points,respectively,under the FA2,FA4,and FA8 treatments relative to the CK.Soil microbial diversity was significantly higher than that of CK after the application of BFA,but the FA8 treatment was lower than the FA4 treatment.The Na⁺/K⁺ ratio in both shoots and roots of maize was lower under FA2 and FA4 treatments than under the CK treatment,while the FA8 treatment increased the Na⁺/K⁺ ratio in the shoots.Maize biomass significantly increased in the mid-to-late growth stages under FA2 and FA4 treatments,while biomass significantly decreased under the FA8 treatment.In summary,the application of 2 g/kg or 4 g/kg of biochemical fulvic acid can positively reduce the alkalinity of soda saline-alkali soil,increase the content of available phosphorus in the soil,improve soil structure,improve soil microbial diversity,and enhance maize salt tolerance and biomass.However,exceeding this dosage range will significantly increase soil salinity and inhibit maize growth.

To clarify the alterations of rice straw decomposition,nutrients release and chemical components in coastal saline paddy soils under different nitrogen (N) application rate,for optimizing the technology of straw returning and realizing the efficient utilization of straw resources in coastal areas.The experimental site was located in Caofeidian District,Tangshan City,Hebei Province.The decomposition characteristics of rice straw and its lignocellulose,as well as the nutrient release characteristics of N,phosphorus (P) and potassium (K) were studied,using a 360-day straw-bag burying method with four different N fertilizer levels,including N0 (0 kg/ha),N1 (225 kg/ha),N2 (300 kg/ha) and N3 (375 kg/ha).Pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) method was used to study the dynamic alterations of principal chemical components of the rice straw residues.The results showed that:the decomposition period of rice straw was divided into three stages,namely,rapid decomposition (0—30 d),slow decomposition (30—210 d) and slow decomposition (210—360 d),and the average decomposition rate of rice straw was 72.5% after 360 days with different N application rates.Increasing N application significantly increased the decomposition rate of rice straw.Compared with the N0 treatment,the N1,N2 and N3 treatments,increased the straw decomposition rate by 6.1, 7.4 and 9.2 percentage points,respectively.The trend of straw carbon (C) release rate was similar to that of straw decomposition rate,while the C release rate was only 43.2% at the end of the experiment.The nutrient release rates of rice straw were as follows:K>P>N.The N,P and K rapid release periods of rice straw was in the 0—30th (38.4%),0—60th (63.7%) and 0—15th (76.7%) days after straw decomposition,respectively.Both N and P of rice straw were enriched during the decomposition period.N application significantly increased the release of N from straw during the decomposition period,P in the early (0—15 d) and late (150—360 d) period,and K in the early period (0—15 d).Compared with the N0 treatment,the N1,N2,and N3 treatments,increased the straw N release by 6.6, 11.1, and 14.7 percentage points,P release by 2.2, 4.0, and 5.6 percentage points,and K release by 1.4, 2.1, and 2.8 percentage points,respectively.The lignocellulose decomposition rates of rice straw were as follows:hemicellulose>cellulose>lignin.Increasing N application significantly promoted the cellulose and hemicellulose decomposition rate of rice straw from day 0—90 and the lignin decomposition rate after day 90.Compared with the N0 treatment,the N1,N2,and N3 treatments,increased the cellulose decomposition rate of rice straw by 5.4, 7.3, and 8.4 percentage points, hemicellulose decomposition rate by 4.9, 6.4, and 7.4 percentage points,and lignin decomposition rate by 2.1, 5.1, and 5.7 percentage points, respectively.2-methoxy-4-vinylphenol,hydroxyacetone,2,3-dihydrobenzofuran,acetosyringone,eugenol,n-hexadecanoic acid,p-methylphenol,2,6-dimethoxyphenol,guaiacol,p-ethylphenol,and stigmasta-3,5-diene were the major (>1% relative) chemical components of straw residues during the decomposition period.Correlation analyses showed that straw decomposition rate,C release rate and cellulose,hemicellulose and lignin decomposition rates,were significantly positively correlated with eugenol,acetosyringone and 2,3-dihydrobenzofuran,while significantly negatively correlated with hydroxyacetone;straw P release rate was significantly positively correlated with hydroxyacetone and significantly negatively correlated with p-ethylphenol,eugenol and acetosyringone;straw K release rate was significantly correlated with p-ethylphenol,eugenol,acetosyringone and 2,3-dihydrobenzofuran,while significantly positively correlated with hydroxyacetone.In conclusion,increasing N application could promote the decomposition rates of rice straw and its lignocellulosic cellulose,and the nutrients release of straw N,P and K in coastal saline paddy field.The recommended optimal N application rate was 300 kg/ha under straw returning 10 500 kg/ha in coastal saline soils.p-ethylphenol,eugenol,acetosyringone,2,3-dihydrobenzofuran,hydroxyacetone,and stigmasta-3,5-diene could indicate the process of straw decomposition in straw residues.Py-GC-MS technique shows a good capability to monitor the chemical components alterations of straw residues,further deepening the understanding of straw decomposition mechanism.

To investigate the function of the β-glucosidase(βGlu)gene PgβGlu4 from Pyrenophora graminea,which previous studies found to be highly expressed during the infection stage,we constructed a subcellular localization vector of PCE2-EGFP-PgβGlu4 and transformed rice protoplasts,observed the fluorescence distribution and analyzed its location of existence.Simultaneously,the PgβGlu4 gene RNAi vector was constructed,and QWC protoplasts were prepared by CaCl₂-PEG4000 mediated method for genetic transformation.The function of PgβGlu4 gene was studied by detecting the vegetative growth and pathogenicity of the RNAi mutants.Phylogenetic analysis of PgβGlu4 and other homologous proteins from different pathogens showed that PgβGlu4 had a closer evolutionary relationship with that from Pyrenophora tritici-repentis.The subcellular localization results showed that PgβGlu4 was mainly localized in the nucleus and cell membrane.Four PgβGlu4 gene RNAi mutants were verified by hygromycin.qRT-PCR analysis showed that the expression of PgβGlu4 gene in four RNAi mutants decreased by 66.31%,68.60%,54.37% and 69.89%,respectively,compared with the wild isolate.The colony diameter was smaller than that of the wild isolate,and their incidence rate was reduced by 56.69,52.76,47.43,and 53.30 percentage points.After infection with the mutant strain of RNAi-PgβGlu4,the relative chlorophyll content in barley leaves ranged from 30.3 to 35.0,which was significantly higher than that of the wild-type group.The effect of PgβGlu4 gene silencing on the height of barley plants before and after infection was significant compared with that of the wild-type.The results indicated that the PgβGlu4 gene was involved in the regulation of the growth,development,and pathogenicity of Pyronophora graminea.

Soybean mosaic virus(SMV)disease can cause significant yield losses and quality deterioration in soybeans,and breeding resistant cultivars remains the only effective strategy for SMV control.Identifying the functional genes associated with SMV resistance provides essential genetic resources for developing resistant varieties.Six MATE candidate genes involved in SMV resistance were identified using a near-isogenic line(NIL)of the qTsmv-3 locus and a transgenic Arabidopsis thaliana plant.A total of 128 MATE family genes were predicted in the soybean genome,which were classified into five subfamilies.Notably,all six MATE candidate genes located at the qTsmv-3 locus clustered within subfamily Ⅰ,exhibiting significant differences in expression levels and tissue specificity based on public data.Among them,Glyma.03G005600 showed the highest expression in aerial tissues(leaves and stems),while Glyma.03G005800 was predominantly expressed in underground tissues(roots and nodules).Following SMV inoculation,the resistant NIL(#NIL-NC)exhibited a 70% reduction in viral accumulation compared with the susceptible line(#NIL-SMC).Concurrently,the expression levels of GmICS1 and GmPR1,key genes in the salicylic acid(SA)-mediated defense pathway,were upregulated by 2.40,15.16 folds,respectively,in #NIL-NC,indicating that qTsmv-3 confers resistance through SA-dependent signaling.Among the 6 MATE candidate genes,only Glyma.03G005300 and Glyma.03G005600 displayed significant differential expression between NILs,which were down-regulated by 61.0% and 82.1%,respectively.Considering their expression patterns and responses to SMV infection,Glyma.03G005600 was identified as the most promising candidate gene for qTsmv-3.Further,the expression of GmICS1 and GmPR1 in transgenic Arabidopsis thaliana(OE_MATE),which carrying Glyma.03G005600,was significantly up-regulated by 4.22,9.12 folds compared with that of wild type(WT)after UV-B stress.These results strongly indicated that Glyma.03G005600 could significantly enhance or affect the expression of genes in salicylic acid signaling pathway,and preliminarily confirmed that Glyma.03G005600 was a key regulatory gene for qTsmv-3 locus.In all,the results laid a foundation for cloning the key genes regulating SMV resistance and provided gene resources for genetic improvement of SMV resistance in soybean.

To address issues such as shallow soil layers,upward movement of the plowpan,and soil salinization in the saline-alkali soils of the West Liaohe Plain,field trials were conducted in Huatugula Town,Horqin Zouyi Middle Banner,Tongliao City,Inner Mongolia Autonomous Region,from 2020 to 2021.Two tillage methods(traditional rotary tillage and powder ridge plowing),two irrigation quotas(2 100,2 700 m³/ha),and mulching and shallow burial measures were set up,resulting in six experimental treatments:2 100 m³/ha irrigation quota+traditional rotary tillage+shallow burial(CK×NM),2 100 m³/ha irrigation quota+traditional rotary tillage+mulching(CK×DM),2 100 m³/ha irrigation quota+powder ridge plowing+shallow burial(FA×NM),2 100 m³/ha irrigation quota+powder ridge plowing+mulching(FA×DM),2 700 m³/ha irrigation quota+powder ridge plowing+shallow burial(FB×NM),2 700 m³/ha irrigation quota+powder ridge plowing+mulching(FB×DM).The effects of powder ridge plowing and mulching treatments on soil properties,structure,saline and alkaline content,and maize yield in the 0—40 cm soil layer under different irrigation quotas were analyzed. The result showed that compared to the CK×NM treatment, in the 0—40 cm soil layer, the soil bulk density decreased by 8.4%—22.9%, the total soil porosity increased by 4.9—14.8 percentage points, and the soil three-phase ratio R value decreased by 34.6%—88.2% under powder ridge plowing + mulching treatment,among them, the bulk density, total porosity, and three-phase ratio R value of the soil treated with FB×DM treatment were significantly reduced by 20.0%,-13.1 percentage points, and 88.2%, respectively;soil moisture content after sowing increased by 5.5—12.1 percentage points in the 20—40 cm soil layer, soil hardness increased by 33.4%—397.5% in the 7.5—17.5 cm soil layer,among them, the soil moisture content, hardness of the FB×DM treatment increased significantly by 12.1 percentage points, 214.3%,respectively;CO₂ flux of the FB×DM treatment increased significantly by 496.4%.Compared to the CK×NM treatment, the powder ridge plowing+mulching treatment reduced the soil pH value, total alkalinity, electrical conductivity, and total salt content in the 0—40 cm soil layer, with reduction rates of 0.7%—10.9%, 2.5%—67.5%, 24.3%—68.7%, and 10.3%—81.0%, respectively. Among them, the soil pH value, total alkalinity, electrical conductivity, and total salt content of the FB×DM-treated soil were significantly reduced by 10.9%, 48.2%, 59.2%, and 80.0%, respectively.Maize germination rate, ear fresh weight, and yield were increased by 13.2—20.1 percentage points,52.5%—68.2%, and 22.4%—45.5%, respectively,among them, the germination rate, ear fresh weight, and yield of the FB×DM treatment were significantly increased by 20.1 percentage points,68.2%, and 45.5%, respectively, compared to the CK×NM treatment. Considering the comprehensive improvement effects and maize yield, it is concluded that the 2 700 m³/ha irrigation quota+powder ridge plowing+mulching(FB×DM) is a more suitable cultivation mode for saline and alkaline land in the West Liaohe Plain.

Metacaspase (MC) belongs to arginine/threonine specific protease,studies have shown that it plays a role in programmed cell death.To investigate the distribution of MC family genes in the genome of Brassica napus and whether they respond to drought stress,this study systematically analyzed the physicochemical properties,phylogeny,gene structure,conserved domains,cis-acting elements,and expression patterns of MC family genes under drought(PEG6000)and abscisic acid(ABA)stress in B.napus.A total of 25 BnMC genes were identified.Chromosomal localization showed that the 25 BnMCs were distributed on 13 chromosomes.Subcellular localization prediction showed that 17 members of the BnMC family were localized in the nucleus and seven members were in the cytoplasm.The phylogenetic tree classified BnMC into two major classes (Type Ⅰ and Type Ⅱ) and four branches (Group A,Group B,Group C,and Group D).BnMCs of the same branch had similar gene structure and conserved motif distribution.The core promoter regions of BnMC contained four types of cis-acting elements:light response element,phytohormone response element,plant growth and development response element and stress response element.Among all the cis-acting elements related to abiotic stress responses,the abscisic acid response element (ABRE) was the most abundant,with a total of 79.All members contained this cis-acting element.The transcriptome sequencing revealed that the expressions of BnMC10,BnMC22,BnMC1,BnMC12 and BnMC8 were up-regulated and the expressions of BnMC4 and BnMC5 were down-regulated after drought treatment.The qRT-PCR assay showed BnMC10,BnMC8,BnMC1 and BnMC12 genes were expressed in both roots and leaves and were up-regulated by both PEG6000 and ABA,with BnMC1 showing the most significant up-regulating changes.In summary,the response of B.napus to drought stress involves the regulation of the expression level of MC family genes.

The soybean germination stage is greatly affected by low-temperature stress,which can have a significant impact on yield.In order to explore genes related to the response of soybean germination to low-temperature stress and investigate the biological processes underlying soybean germination tolerance to cold,this study conducted transcriptome sequencing on seeds germinating for three days from eight materials showing significant differences in low-temperature tolerance during germination.Differential expressed genes (DEGs) between materials tolerant and sensitive to low temperatures were identified and subjected to GO enrichment analysis,KEGG pathway enrichment analysis,and transcription factor analysis.Among 231 DEGs identified in 15 contrasting groups of low-temperature tolerance,159 DEGs were up-regulated and 72 DEGs were down-regulated in cold-sensitive soybeans.GO enrichment analysis revealed that DEGs were mainly involved in biological processes such as cellular processes (GO:0009987),metabolic processes (GO:0008152),biological regulation (GO:0065007),response to stimulus (GO:0050896),binding (GO:0005488),transporter activity (GO:0005215),and transcription regulator activity (GO:0140110).KEGG pathway enrichment analysis indicated that DEGs were significantly enriched in starch and sucrose metabolism pathways (ko00500).Genes involved in seed development (Glyma.03G144400, Glyma.19G147200,Glyma.10G027600,Glyma.10G247500,Glyma.20G147600),metabolic reactions (Glyma.05G004300,Glyma.17G086400),and genes encoding glutathione oxidase (Glyma.01G219400) were up-regulated in cold-sensitive materials.Fifteen transcription factors from families such as MYB,AP2/ERF,and NAC were identified among the 231 differentially expressed genes,suggesting that soybeans respond to low-temperature stress during germination by regulating various biological processes,metabolic pathways,and signal transduction pathways.

In order to explore the effect of straw interlayer on soil respiration and the chemical structural stability of organic carbon components during the remediation of saline-alkali soil,two experimental treatments were set up in the alfalfa-planting farmland at the Saline-Alkali Soil Improvement Experimental Demonstration Base in the Agricultural High-Tech Zone of Dongying City,Shandong Province:a straw interlayer treatment (S,with a 5 cm thick straw layer buried at a 35 cm depth) and a control (CK,without an interlayer). Soil respiration characteristics of the different treatments were analyzed,and comprehensive discussions were conducted in conjunction with soil pH,electrical conductivity(EC),organic carbon component content,and chemical structure characteristics of the soil profile.The results showed that:compared with CK,the S treatment significantly increased the soil respiration rate during the alfalfa growth period, with a maximum increase of 79.84%. Furthermore, the S treatment reduced soil EC in the straw interlayer (35—40 cm) and the overlying soil layer (0—35 cm), effectively inhibiting the upward migration of salt. The 40—50 cm soil layer was identified as a critical zone for organic carbon transformation. In this layer, the S treatment significantly increased SOC content (by 16.67%) and highly significantly elevated particulate organic carbon (POC) and dissolved organic carbon (DOC) contents (by 208.07% and 83.41%, respectively) compared with CK. Characterization of the molecular structure of organic carbon in the 40—50 cm layer revealed distinct responses. For DOC, the S treatment reduced the magnitude-weighted averages of unsaturation (by 30.60%) and aromaticity index (by 4.84%) compared with CK, decreasing the proportion of unstable carbon. For POC, the S treatment increased the relative abundances of alkyl C(10.32 percentage points)and O-alkyl C (8.39 percentage points) while reducing carboxyl C (14.24 percentage points), thereby enhancing POC structural stability. However, for bulk SOC, the S treatment decreased the proportions of alkyl C(3.14 percentage points) and aromatic C(3.38 percentage points) while increasing O-alkyl C(5.17 percentage points)and carboxyl C(1.56 percentage points). This shift indicated a decrease in recalcitrant carbon and an increase in labile carbon, resulting in reduced SOC structural stability. Correlation analysis showed that the significant increased in soil respiration was highly significantly and significantly positively correlated with POC and SOC contents in the critical soil layer, respectively. Specifically, the accumulation of O-alkyl C(a labile component) in the chemical structure of SOC reduced the stability of organic carbon,which was the main reason for the increase in soil respiration.In conclusion,incorporating a straw interlayer significantly increased soil respiration in the short term,which was closely related to the reduction in the stability of the organic carbon chemical structure in the key soil layer.Under future "carbon neutrality" strategies,research on the selection of interlayer materials should be considered.

To elucidate the role of WRKY transcription factor family members in the dynamic regulation of wheat growth and development,as well as in responses to abiotic stresses,this study investigated the expression patterns of TaWRKY gene under drought,high salinity,and low-temperature stress conditions.Using the common wheat cultivar Chinese Spring as the experimental material,we obtained the TaWRKY70 gene through molecular cloning.The coding sequence of TaWRKY70 was 885 bp in length,encoding a 294-amino-acid hydrophilic and unstable protein.Bioinformatics analysis revealed that the protein possessed a typical WRKYGQK conserved domain and a C₂HC-type zinc finger structure,classifying it as a Group Ⅲ WRKY transcription factor.Cis-regulatory element analysis of the TaWRKY70 promoter region identified regulatory elements involved in responses to methyl jasmonate,abscisic acid,and ethylene.Co-expression gene analysis suggested that TaWRKY70 was associated with multiple stress response processes in wheat,including hormone signaling,defense against microbial pathogens,and responses to cold stress.Phylogenetic analysis indicated that TaWRKY70 shared a close evolutionary relationship with WRKY70 proteins from other Poaceae species,such as barley,maize,sorghum,and foxtail millet.Subcellular localization experiment further confirmed that TaWRKY70 was localized in the nucleus,consistent with the characteristics of a transcription factor.Expression pattern analysis showed that TaWRKY70 was expressed in wheat roots,stems,leaves,young spikes,and grains,with higher expression levels observed in roots and leaves.Under abiotic stress conditions,TaWRKY70 expression was downregulated in response to abscisic acid and low-temperature treatments but upregulated under salicylic acid,NaCl,PEG6000,and high-temperature treatments.In conclusion,the cloning of TaWRKY70 gene and analysis of its expression pattern provide a basis for the next step to analyze the molecular mechanism of TaWRKY70 involved in wheat stress resistance.

Sunflower is an important oil crop with strong salt tolerance in China.However,the production of sunflower is severely restricted by the intensification of salinization in main producing areas.To understand the salt tolerance mechanism of salt tolerance-related traits at the seedling stage of sunflower,and to identify SNP loci and candidate genes significantly associated with salt tolerance at the seedling stage of sunflower,124 sunflower germplasm accessions were used as materials.Under the stress of 250 mmol/L NaCl,genome-wide association analysis and mining of salt-tolerance loci/genes were performed based on 5 traits including plant height,leaf area,aboveground fresh weight,underground fresh weight,and SPAD value.The results showed that after 14 days of salt stress,the five trait indexes of relative plant height,relative leaf area,relative fresh weight of the above-ground part,relative fresh weight of the underground part and relative SPAD value at seedling stage had significant variations and normal distributions,and the most obvious influences were the relative fresh weight of the underground part and relative leaf area (with a coefficient of variation of over 46.57%).Eighteen significantly-associated SNP loci were obtained by genome-wide association analysis,among which Chr35448-18789497 was detected in several salt-tolerant traits;gene searches were conducted at 80 kb each in the upstream and downstream of associated loci,45 related candidate genes were screened out.Through scanning and gene annotation analysis of the associated locus intervals,four candidate genes that might be related to the salt tolerance trait of sunflower seedlings were discovered.A genome-wide association analysis was conducted using the phenotypes of salt tolerance identification at the seedling stage and genomic SNP data for sunflower germplasm accessions.SNP loci significantly associated with salt tolerance at the seedling stage were detected,and candidate genes related to salt tolerance at the seedling stage of sunflower were selected,which laid the foundation for the subsequent verification of salt-tolerant genes.

Although high-throughput KASP markers have been developed for the wheat quality subunit 7^OE,they are different from the KASP markers developed by SNP,the problem of not being able to effectively distinguish between homozygous and heterozygous remains.To clarify the issue of whether the 7^OE subunit is homozygous,this study used Jinqiang 6(containing 7^OE+8^* subunits)and Kenong 199(containing 7+9 subunits)and hybrid offspring as materials,and used the Waxy-D1 gene of wheat as an internal reference gene.The relative copy number of the 7^OE gene to the reference gene was detected by quantitative PCR using the universal dual-color fluorescence used in KASP markers to determine whether the 7^OE gene exists and whether it is homozygous,and the detection results were verified by a relevant molecular marker.The results showed that the relative copy number of the parent Jinqiang 6,with the 7^OE gene,was the highest,the relative copy number of the parent Kenong 199,without the 7^OE gene,was 0,and the relative copy number of their hybrid F₁ generation was intermediate,and the three types were easily separated.In its F₂ segregating population,the relative copy numbers of the 7^OE gene were also easily divided into high,medium and 0 three types.The genotypes that were detected as homozygous and heterozygous for the 7^OE gene were further detected by the PCR marker of the 9 subunit(which can detect 9 subunit and the 8^* subunit that are closely linked to the 7 subunit and the 7^OE subunit,respectively),and the results were completely consistent.The high-throughput 7^OE universal dual-color fluorescence quantitative PCR marker established in this study can accurately distinguish whether the 7^OE subunit is present,and whether it is homozygous,which has a positive effect on promoting the molecular marker-assisted selection of high-quality subunit 7^OE.

The ubiquitination pathway is one of the key signaling pathways in response to drought stress.In order to clarify the biological function of E3 ubiquitin ligase TaSINA101 gene in response to drought stress,the TaSINA61,TaSINA101 and TaSINA105 genes were cloned from JM47,an excellent drought-resistant wheat cultivar,and their sequence characteristics were analyzed by bioinformatics methods,and the expression levels of the three genes in wheat roots and leaves were detected by qRT-PCR under PEG-6000,NaCl,low temperature and ABA treatments.The heterologous expression of TaSINA101 in transgenic rice was used to analyze the biological function of TaSINA101 in response to drought stress.The results showed that the TaSINA61,TaSINA101 and TaSINA105 genes contained one intron and two exons,and the encoded proteins were composed of 282 amino acids.The qRT-PCR expression analysis showed that the expression of these three genes was induced by various abiotic stresses such as drought stress in roots and leaves.Phenotypic analysis of TaSINA101 transgenic rice under drought stress showed that the leaf fresh weight and dry weight, maximum root length, average root diameter and leaf relative water content of transgenic rice lines OE-1, OE-2 and OE-3 were significantly lower than those of wild type,while the relative conductivity of leaves of transgenic rice lines OE-1 and OE-2 was significantly higher than that of wild type.Therefore,TaSINA101 negatively regulates drought stress tolerance in rice.This study provides a basis for in-depth analysis of the biological function of TaSINA101 gene in wheat.

In order to clarify the regulatory network of oil accumulation in Brassica napus and breed oilseed rapeseed varieties with high oil content.The seed transcriptome data of four rapeseed inbred lines at 25,35,and 45 days after flowering were used to identify candidate genes affecting oil content by transcriptome analysis and correlation analysis.Consequently,a total of 1 530 genes were identified exhibiting differential expression across all three period,comprising 986 up-regulated genes and 544 down-regulated genes.GO enrichment analysis of these differentially expressed genes detected 83 lipid biosynthesis genes,79 lipid degradation genes,21 lipid transporter genes and 80 transcription factors.To further analysis of these differentially expressed transcription factors,genes including BnTT8,BnGL2,and BnNAC082 were identified.Combined with the oil content data of 50 semi-winter rapeseed in three different regions over two years,four SNPs were identified in the exons region of the BnNAC082-A03 significantly associated with oil content using genome-wide association studies.Two haplotypes were detected in the region of this gene and BnNAC082-A03_Hap1 corresponding accessions showed significantly higher oil content than that of Hap2.Additionally,it utilized these transcriptomic data to construct co-expression analysis network,and in the sub-network revealed that BnNAC082-A03 was directly connected with BnTT8-A09 and BnGL2-C06,forming a potential molecular regulatory network affecting seed lipid accumulation.

Fusarium head blight(FHB)is a devastating fungal disease that seriously threatens the safety of wheat production.Marker-assisted selection(MAS)and pyramiding of resistance genes represent efficient strategies for FHB-resistant breeding.To establish a high-throughput screening system for FHB resistance genes and enhance wheat resistance in Sichuan Province,we performed genome-wide genotyping using a 100K SNP array on 14 Sichuan wheat varieties(lines)along with three FHB-resistant genetic materials.Based on the reported genetic linkage intervals of major FHB resistance genes(Fhb2,Fhb4,Fhb5),we identified SNPs co-segregating with Fhb5 or linked to Fhb2,Fhb4,and subsequently developed kompetitive allele-specific PCR(KASP)markers.Results showed that the genetic relationship of 17 wheat varieties(lines)could be clustered into two major groups:two northern wheat-derived resistant materials(NMAS070 and NMAS069)formed an independent cluster distinct from the Sichuan varieties(lines)while the remaining 15 varieties(lines)were clustered together and subdivided into two subgroups.Functional gene profiling revealed FHB-resistant parents carried superior resistance loci,whereas agronomic parents harbored favorable alleles for yield and quality traits.Through SNP screening,we identified 8 critical SNPs within the linkage intervals of Fhb2,Fhb4 and the co-segregation region of Fhb5.These SNPs enabled the successful development of 4,2,and 2 high-specificity KASP marker systems for Fhb2,Fhb4 and Fhb5,respectively.Validation experiments confirmed all KASP markers achieved precise genotyping and were effectively implemented in molecular breeding for FHB-resistance.This study established a high-efficiency KASP marker system for Fhb2,Fhb4 and Fhb5,providing a robust technical platform for improving FHB resistance breeding of wheat varieties in Southwest China.

Yellow-seeded rapeseed is popular with consumers because of the appearance and good quality of rapeseed oil.However,the unstable separation of progeny traits seriously affects its large-scale application.In order to analyze the internal reasons for the unstable separation of yellow-seeded rapeseed traits,the differences in the internal physiological mechanisms between yellow and black seeds in yellow-seeded rapeseed were explored.Yellow-seeded rapeseed(CK)was used as the material to study the differences in agronomic traits,physiological and biochemical indexes,and seed coat color-related genes between yellow(Y)and black(B)seed plants in the separation offspring.The results showed that the rhizome diameter and plant height of Y were larger than those of CK and B.The plant height of B was significantly different from that of CK and Y,respectively,and the rhizome diameter of B was significantly different from that of Y.The disease index of Y was 1.97,and the disease indexes of CK and B were 2.55 and 3.33,respectively,indicating that Y was superior to CK and B in disease resistance.The content of MDA in Y leaves was the lowest at 9—10 leaf stage,and the POD activity in Y and CK flowers continued to increase at flowering stage,indicating that yellow-seeded rape had strong stress resistance.The expression levels of TT18 and TT8 genes in B and Y at 7—8 leaf stage and 9—10 leaf stage were higher than those in CK,and the expression levels of TT18 gene in B and Y at final flowering stage were significantly lower than those in CK.At 28 days after pollination,the expression of MYB47 gene in Y seed was the highest,which was 5.56 times that of CK and 5.79 times that of B,respectively.The expression of TT8 gene was the highest in Y at 21 days after pollination,which was 3.30,2.29 times that of CK and B,respectively.It showed that yellow-seeded rapeseed had obvious advantages in oil content and stress resistance.Therefore,vigorous development of yellow-seeded rapeseed can provide new ideas for improving the supply of rapeseed oil and solving the safety of edible oil in China.

This study investigated the impacts of drought on the agronomic,physiological,and biochemical characteristics of peanuts developed a classification methodology for drought resistance,screened the varieties of peanut with drought resistant.Agronomic traits of 27 peanut germplasms under drought stress were measured 30 days after sowing,and drought resistance was graded by correlation analysis,principal component analysis,cluster analysis,and membership function method.Drought-resistant peanut JNH3,intermediate L231,and drought-sensitive L236 were selected for determination of physiological and biochemical indexes and microstructure observation,and different drought-resistant peanut germplasms were identified.The results showed that after drought treatment,the decrease in main stem height ranged from 2.26% to 34.06%,the length of first branches decreased from 1.11% to 57.20%,the main stem base coarse decreased from 1.54% to 38.36%,and the root-shoot ratio decreased from 65.01% to 92.83% under different environments.According to the comprehensive weighted membership function,peanut materials were grouped into three types:drought-resistant,intermediate,and drought-sensitive.Among them, the above-mentioned traits of the intermediate type and the drought-sensitive type peanuts reached a significant level compared with the control. Under drought stress,ROS in functional leaves of peanut increased,and different expressions of ROS were found by NBT and DAB staining,among which NBT and DAB staining were JNH3,L231,and L236 from light to deep,conforming to the classification of drought resistance type.Compared with the control,the POD activity of JNH3,L231,and L236 increased by 42.71%,26.04% and 20.59% respectively,among different varieties,and CAT activity trend was consistent with the above.The SOD activity of JNH3,L231,and L236 increased by 48.01%,63.49% and 73.15% respectively,among different varieties,and the MDA activity trend was consistent with the above.The proline content of JNH3,L231,and L236 was increased by 1.1,1.07 and 1.03 times,and the soluble sugar content of JNH3,L231,and L236 was increased by 44.06%,31.54%,and 38.62% respectively,among different varieties.Under drought stress,peanut root growth was limited,the total root length and total root area were significantly reduced,the root tip cells were partially necrosis,and the degrees were shallow to deep in JNH3,L231,and L236 respectively.

The effects of castor cake meal fertilizer on soil fertility and peanut yield and quality were studied.Using the peanut variety Silihong in Northeast China as the experimental material,a two-year field positioning experiment(2022 and 2023)was carried out,with no fertilization(CK),castor cake meal fertilizer(B1:2 520 kg/ha,B2:5 040 kg/ha,B3:10 080 kg/ha),chemical fertilizer(F1:175 kg/ha,F2:350 kg/ha,F3:700 kg/ha),cow manure(N1:3 724 kg/ha,N2:7 448 kg/ha,N3:14 896 kg/ha).A total of 10 treatments were used to analyze the effects on soil fertility and peanut yield and quality.The results showed that the application of castor cake meal fertilizer in 2022 and 2023 could improve soil nutrients to varying degrees.B3 treatment could significantly reduce soil pH value,and the effect of soil organic carbon,total nitrogen,total phosphorus,total potassium,alkali-hydrolyzed nitrogen,basic phosphorus,and available potassium content were the best.Compared with CK,the average increase was 67.58%,64.56%,70.55%,11.33%,75.76%,149.97% and 116.84%,respectively.Compared with CK,chemical fertilizer and cow manure made soil organic carbon,total phosphorus and basic phosphorus content increase by an average of 5.94% and 11.48%,16.67% and 16.67%,33.35% and 23.94%,respectively.The full fruit weight per plant,100 fruit weight,100 grains weight of peanut with B3 treatment were better than that treated with chemical fertilizer and cow manure,which increased by an average of 105.43%,127.91%,19.54% and 22.75%,16.79%,24.17%,respectively.B3 treatment had the highest peanut yield,which was 1 876.22 kg/ha higher in 2023 than 2022.B1 treatment increased the contents of peanut fat and oleic acid and decreased the content of linoleic acid and palmitic acid,compared with CK,the content of peanut fat and oleic acid increased by 6.07 and 4.23 percentage points,and the contents of linoleic acid and palmitic acid decreased by 1.79 and 0.89 percentage points.The contents of soil organic carbon,total nitrogen and alkali-hydrolyzed nitrogen were positively correlated with peanut fat content.The contents of soil organic carbon,total nitrogen and alkali-hydrolyzed nitrogen were significantly positively correlated with peanut yield.In conclusion,castor cake meal fertilizer could improve soil,increase peanut yield and quality.

To elucidate the molecular mechanisms underlying peanut pod responses to water stress,this study employed pot experiments combined with transcriptomic analysis.Using well-watered conditions as the control, we systematically investigated the effects of periodic drought and waterlogging stress during the flowering-pegging stage on yield,quality,and gene expression in peanut pods. Results demonstrated that both drought and waterlogging stresses significantly reduced peanut pod yield(by 26.43% and 77.69%,respectively)and crude fat content in kernels (by 9.46, 6.71 percentage points,respectively).Transcriptomic analysis further revealed 1 525 and 1 382 differentially expressed genes(DEGs)in the drought-stress and waterlogging-stress groups compared to the control, respectively, with down-regulated expression being predominant in both sets of DEGs. Specifically, drought stress suppressed six key metabolic processes related to lipid and fatty acid metabolism in peanut pods,with 88.38% of associated genes showing downregulated expression,indicating that lipid metabolic disruption may be the primary cause of yield and quality reduction under drought. Waterlogging stress predominantly interfered with pod metabolism and defense functions by downregulating genes associated with catalytic activity,transmembrane transport,redox reactions,and biosynthesis of secondary metabolites.Moreover, KEGG enrichment analysis indicated that metabolic pathways and biosynthesis of secondary metabolites were significantly affected under both water stress conditions. Key gene validation via qRT-PCR corroborated the RNA-seq data, confirming the reliability of the transcriptomic findings. In summary, this research elucidates the molecular basis of peanut pod response to water stress at the transcriptome level, demonstrating that lipid metabolic disruption is the primary factor underlying yield reduction and quality deterioration under drought stress, whereas peanut pods mitigate the adverse effects of waterlogging mainly by modulating the expression of genes associated with redox homeostasis and metabolic pathways.

In order to cultivate good quality rice,it utilized 139 rice germplasm resources from home and abroad as materials to analyze the total protein content of rice grains in 2022-2023,combined with 255 501 SNP markers obtained from whole-genome sequencing(depth of coverage of 10×),and performed genome-wide linkage analysis by using a general linear model to avoid the influence of false positives to select the genes with the highest thresholds for haplotype analysis.The genes related to the total protein content of rice grains were predicted based on the results of previous studies and gene function annotation,and the relative expression of the predicted genes was analyzed by Real-time Fluorescence PCR.The results showed that the total protein content of 139 rice seeds belonged to moderate variation,with coefficients of variation of 21.66% and 20.65%,respectively,which conformed to the normal distribution.Through genome-wide association analysis,a total of 55 significant SNPs were obtained in both environments,distributed on chromosomes 1,2,4,5,6,8,11,and 12,of which 16 consecutive and with upstream and downstream intervals of no more than 100 kb SNPs were distributed on chromosome 11.Further haplotype analysis of genes with strong correlation between the upstream and downstream intervals within 50 kb(±50 kb)of the loci of significant SNPs on chromosome 11 was conducted,combined with the results of functional annotation of the genes and the analysis of the relative expression of the seed grain at the irrigating stage,we preliminarily hypothesized that LOC_Os11g08460 was associated with the total protein content of the seed grain of rice,which encodes the Dnak/Hsp70s protein family.In conclusion,candidate gene prediction and haplotype analysis of total protein content of 139 rice germplasm resources using genome-wide association analysis can provide new genes for genetic improvement of rice quality and accelerate the process of rice improvement.

The aims were to study the effect of maize straw returning on soil fertility and productivity under wheat-maize rotation system in North China,and to explore the best way and suitable amount of maize straw returning.From 2021 to 2023,a field split-plot experiment was conducted in Quzhou County,Hebei Province to compare the effects of different maize straw returning methods and amounts on wheat growth,yield composition and soil organic matter content.There were two treatments in the main area,which were cutting and crushing.The sub-area was the amount of straw returning to the field,4 levels,which were 0,0.5,1.0,and 1.5 times of the amount of maize straw in the year.The results showed that compared with straw crushing,the yield of wheat increased by 4.3% under straw cutting treatment,the number of spikes increased by 7.6%,and the harvest index increased by 1.4%,with all significant difference in 2023.The N content of wheat straw was significantly increased by 0.04 percentage points,the N and P uptake of wheat aboveground were significantly increased by 10.8% and 14.3%,respectively(2022),but the K content of wheat straw was significantly reduced by 0.13 percentage points(2023).Cutting treatment could significantly promote the growth of wheat before and after winter,and the NDVI value at jointing stage significantly increased by 11.9%(2022).Soil pH increased by 0.22 units(2023).With the increase of maize straw returning amount,wheat yield and NDVI value at jointing stage showed parabolic model of first increased and then decreased,the N,P and K absorption of above ground wheat showed a significant downward trend,and soil organic matter showed a continuous significant increase trend.Based on the growth potential and yield of wheat,the suitable returning ratio of straw cutting treatment was 58%—62%,and the suitable returning ratio of crushing treatment was 29%—42%.Under the conditions of this experiment,maize straw cutting returning has obvious advantages in promoting wheat growth,and increasing wheat yield.It can be popularized and applied in wheat-maize rotation areas that 58%—62% maize straw is returned into the soil.

Wheat stripe rust,caused by Puccinia striiformis f.sp.tritici,is a major disease that severely threatens China's food security.Urediniospores are key agents for the reproduction,dissemination,and infection of the pathogen,but the regulatory mechanisms of sporulation-related genes remain unclear.This study aims to screen and validate the function of the candidate gene PsCON6,which is highly expressed during the early infection stage of P.striiformis,to provide new insights into its pathogenic mechanisms.The PsCON6 gene was obtained from P.striiformis via homologous cloning,and its expression pattern during early infection was analyzed using qRT-PCR.Bioinformatics technology analysis of the amino acid sequence,conserved domains and physicochemical properties of the PsCON6 protein.Barley Stripe mosaic virus host-induced gene silencing(BSMV-HIGS)was used to transiently silence PsCON6,followed by measurements of host reactive oxygen species accumulation,fungal hyphal length and area,and pathogen biomass.Subcellular localization of PsCON6 was determined through transient expression assays.PsCON6 was significantly upregulated during the early infection stage of P.striiformis.The encoded protein contained two conserved conidiation-specific protein 6 domains and consisted of 83 amino acids.After HIGS-mediated silencing of PsCON6,the level of reactive oxygen species in the host significantly increased,while the length and area of the fungal hyphae significantly decreased,whereas urediniospore production remained unaffected.Subcellular localization revealed that PsCON6 was localized to the cell membrane.PsCON6 likely participates in regulating hyphal growth in P.striiformis but does not directly influence urediniospore formation,suggesting potential functional redundancy.The research findings provide new targets for revealing the pathogenic mechanism of wheat stripe rust and lay the foundation for further in-depth analysis of its molecular mechanisms.

To explore the disparities in overwintering rates and yields between Brassica napus L.and Brassica rapa L.in the cold and arid regions of the north,as well as to examine the correlation between diverse agronomic traits,yield,and cold resistance-enhancing traits,it utilized 45 B.napus L.and 22 B.rapa L.varieties as subjects.It conducted a statistical analysis of their overwintering rates,average yields and agronomic traits.It also studied seeding density experiments with B.napus L.and B.rapa L.varieties that exhibited similar yield levels,compared root traits between B.napus L.and B.rapa L..The results indicated that among the 67 varieties tested,the average overwintering rate of B.rapa L.(97.59%) was significantly higher than that of B.napus L.(65.87%).The average yield potential of B.napus L.was higher than that of B.rapa L..For B.rapa L.,which was capable of stable overwintering,increasing the seeding density significantly elevated the number of effective plants but did not augment the average yield.Conversely,for B.napus L.,its lower overwintering rate constrained the increase in the number of effective plants,thereby limiting the average yield.B.napus L.with a high overwintering rate exhibited agronomic traits such as higher branching positions and angular density,fewer secondary branches,total branches,and effective siliques per plant after overwintering.Comparison of root traits between B.napus L.and B.rapa L.showed that enlarged root systems,short hypocotyls,and growth points positioned below the ground surface were advantageous traits contributing to the robust cold resistance of B.rapa L..It proposes that breeding density-tolerance B.napus L.with root traits akin to those of B.rapa L.and with growth points situated below the ground surface represents a pivotal breeding direction for enhancing the overwintering rate of B.napus L.,fostering increased rapeseed yield,and ensuring oil supply stability.

This study explored the effects of full-width uniform seeding on the canopy light energy utilization characteristics,dry matter accumulation and transport of winter wheat in saline-alkali land,and clarified the physiological mechanism of high yield and high efficiency,to provide theoretical and practical basis for the promotion of full-width uniform seeding of winter wheat in the Yellow River Delta.In the growing season of winter wheat from 2022 to 2024,Jingyou 368 wheat variety was used as the material,and two seeding patterns of full-width uniform seeding and conventional drill seeding were set up.The differences of yield,dry matter accumulation,dry matter transport,canopy photosynthetically active radiation interception amount and radiation use efficiency under different seeding patterns were analyzed,and the correlation analysis was carried out.The results showed that the yield and spike number of wheat under full width uniform seeding were higher than those under conventional drilling seeding.The full-width uniform seeding achieved extremely significant increases of 18.35% and 46.97% from 2022 to 2023,and 18.71% and 47.21% from 2023 to 2024,respectively.Under the full-width uniform seeding,the wheat stem & tillers number was higher than that under the conventional drilling seeding.From 2022 to 2023,full-width uniform seeding significantly increased the tiller number by 58.83%.From 2023 to 2024,full-width uniform seeding extremely significantly increased the tiller number by 57.30%.The dry matter accumulation of wheat at anthesis stage,dry matter accumulation at maturity stage and dry matter translocation of vegetative organs before anthesis under full-width uniform seeding were higher than those under conventional drilling seeding.From 2022 to 2023,full-width uniform seeding achieved extremely significant increases of 75.78%,41.70% and 109.69%,respectively,and from 2023 to 2024,full-width uniform seeding achieved extremely significant increases of 71.23%,40.81% and 98.07%,respectively.The leaf area index,canopy photosynthetic active radiation interception and radiation use efficiency of wheat under full-width uniform seeding were higher than those of conventional drilling seeding.The full-width uniform seeding in 2022—2023 achieved extremely significant increases of 58.36%,4.11% and 47.17%,respectively,and the full-width uniform seeding in 2023—2024 achieved extremely significant increases of 59.78%,4.11% and 44.00%,respectively.In summary,the full-width uniform seeding of wheat in saline-alkali land improves the canopy light energy utilization performance and tiller productivity by shaping a reasonable population structure and improving the seedbed environment,which is conducive to the formation of plant photosynthetic products and the increase of spike number per unit area,and ultimately achieves high yield of wheat.Therefore,full width uniform seeding is a better seeding pattern for stable and high yield of winter wheat in saline-alkali land of the Yellow River Delta.

The aim is to establish a rapid and accurate method for detecting Avian hepatitis E virus(aHEV)antibodies.The ORF2 protein of the CaHEV-GDSZ01 strain was expressed in an E.coli prokaryotic expression system and utilized as the coating antigen.The reaction conditions were optimized to determine the optimal working conditions,leading to the establishment of an indirect ELISA method for detecting aHEV chicken serum antibodies.The specificity,sensitivity and repeatability of the method were tested,and compared with Western Blot method,chicken clinical serum samples were preliminarily detected.The results showed that ORF2 protein was successfully expressed and purified,and Western Blot assay confirmed that ORF2 protein could specifically react with aHEV-positive chicken serum.The optimal reaction conditions for ELISA were as follows:ORF2 protein was coated with 200 ng per well at 4 ℃ overnight(12 h);blocked with 5% nonfat milk at 37 ℃ for 1-2 h;serum was diluted at 1∶1 600,incubated at room temperature for 1 h;rabbit anti-chicken IgY-HRP was diluted at 1∶5 000,incubated at 37 ℃ for 90 min;TMB was incubated at 37 ℃ for 20 min.The ELISA method could detect the positive sera of aHEV diluted 51 200 times,and did not cross-react with positive sera of other chicken-borne viruses such as NDV,AIV-H5,AIV-H9,ALV,REV,and MDV.The intra-batch and inter-batch coefficients of variation were both less than 10%,rate between the Western Blot method was 95.12%. Clinical serum samples from chicken farms in different areas of Guangdong were tested by this ELISA method, and the total positive rate reached 66.34%.In conclusion,this study successfully established a simple,effective,specific,sensitive and reproducible serological detection method for aHEV infection,which can provide technical support for the surveillance,prevention and control of aHEV infection at grassroots.

To investigate the effects of climate warming and combined application of organic and inorganic fertilizer on soybean growth and yield,field experiments were conducted using infrared radiation warming systems to simulate temperature elevation.The experimental design included two temperature levels(T0:Ambient temperature;T1:Warming by 2.9 ℃)and two fertilization regimes(SF:Sole inorganic fertilizer;OF:Combined organic-inorganic fertilizer),aiming to examine the impacts of warming and fertilization practices on soybean dry matter accumulation,photosynthetic rate,leaf enzyme activities,and yield.Results demonstrated that compared to ambient temperature,warming shortened the pre-flowering and post-flowering growth periods by 3—4 d,5—6 d,respectively.Warming significantly reduced leaf nitrate reductase(NR)and glutamine synthetase(GS)activities during the pod-filling stage,decreased leaf area index(LAI)by 9.0%—13.7%,and suppressed net photosynthetic rate,collectively leading to 4.1%—19.3% reductions in post-flowering plant height and aboveground dry matter accumulation.In contrast,the combined organic-inorganic fertilization enhanced NR and GS activities,improved post-flowering photosynthetic efficiency,promoted plant growth,and increased the aboveground dry matter accumulation by 4.1%—15.3% compared to sole inorganic fertilization,with a significant interaction observed between NR and alanine aminotransferase activities.Yield analysis revealed that warming caused 9.7%—16.6%,13.3%—19.0% declines in pods per plant and grains per plant,respectively,resulting in 13.7%—21.1% yield reductions.Conversely,organic-inorganic fertilization increased grains per plant by 12.5% and pods per plant by 9.9%—10.5%,achieving 7.6%—10.8% yield improvements.Notably,significant positive correlations were detected among LAI,photosynthetic rate,nitrogen metabolism enzyme activities,and final yield.These findings demonstrate that climate warming inhibits post-flowering photosynthetic efficiency and aboveground growth in soybeans,while integrated organic-inorganic fertilization partially mitigates warming-induced yield losses through enhancing enzyme activities and photosynthetic performance,providing critical insights for adaptive agricultural practices under global warming scenarios.

This study explored the relationship between cotton GhERF14 gene and Fusarium oxysporum pathogenicity,analyzed the molecular mechanism of F.oxysporum pathogenicity,and tentatively explored the response of cotton GhERF14 gene to Fusarium wilt disease and its regulatory effect on related resistance genes,to provides some theoretical basis for breeding new cotton cultivars resistant to wilt.Gene cloning and virus-induced gene silencing(VIGS)were used to construct the non-conserved domain interference vector pTRV2-GhERF14.Using Real-time fluorescence quantification(qRT-PCR)technology and VIGS technology,the expression characteristics of GhERF14 and downstream genes related to lignin,ethylene(ET),jasmonic acid(JA),salicylic acid(SA),antioxidant enzymes and disease progression-related protein(PR)were analyzed after F.oxysporum stress and hormone treatment,and the role of GhERF14 in the process of cotton disease resistance was analyzed.The results indicated that inhibition of GhERF14 gene expression could significantly reduce the synthesis of jasmonic acid(JA),salicylic acid(SA)and ethylene(ET)and the expression of genes related to the signaling pathway.After GhERF14 gene silencing by VIGS technology,cotton plants were more susceptible to Fusarium wilt.These results suggested that GhERF14 may play an important role in the pathogenesis and host-pathogen interaction of F.oxysporum.

To explore the effects of nitrogen application rate on nitrogen uptake and translocation characteristics,nitrogen use efficiency,and the formation mechanisms of yield and quality in soft wheat under weak light stress during the after anthesis stage,under pot conditions,the soft wheat variety Quanmai 725(QM725)was used as the material,and the ¹⁵N tracer method was used.Two nitrogen rates were set in the experiment,namely N1(N 120 kg/ha),N2(N 180 kg/ha),and two shading treatments were set at the wheat filling stage(7-35 d after anthesis)under each nitrogen application rate,namely CK(no shading),SH(30% shading).The relationship between nitrogen application rate and grain yield and quality of soft wheat under weak light after anthesis was analyzed,and the effects of different nitrogen application rates on nitrogen accumulation,transport,grain yield and quality of soft wheat under weak light after anthesis were studied.The results showed that compared with the control,under the conditions of N1 and N2,the nitrogen accumulation of plants at flowering stage and vegetative organs at maturity stage was significantly reduced by weak light treatment after anthesis,and the proportion of nitrogen from fertiliser was significantly higher than that from soil nitrogen,while the proportion of nitrogen accumulation in grains at maturity stage from soil nitrogen was significantly higher than that from fertiliser nitrogen.Under the same nitrogen application rate,the proportion of basal nitrogen fertiliser was greater than that of topdressing nitrogen fertiliser.Under the same weak light treatment conditions,compared with N1,N2 increased the accumulation of fertiliser nitrogen at flowering stage,the accumulation of total nitrogen and fertiliser nitrogen at maturity stage,and the accumulation of total nitrogen,fertiliser nitrogen and soil nitrogen at maturity stage.Under N1 and N2 treatments,the nitrogen harvest index,nitrogen harvest index,nitrogen production efficiency,grain number per spike,1000-grain weight and grain yield of wheat decreased significantly with the decrease of light intensity after anthesis.The content and accumulation of protein and starch in soft wheat grains increased significantly with the increase of nitrogen fertilizer.However,under the same nitrogen application rate,weak light stress reduced the starch content,protein and starch accumulation in grains.Weak light stress after anthesis significantly affected the nitrogen accumulation of soft wheat plants,reduced the transport efficiency of storage substances from vegetative organs to grains after anthesis,resulting in a decrease in the contribution rate of vegetative organs to grains,which was not conducive to the overall nitrogen transport efficiency of plants.With the increase of nitrogen application rate,the nitrogen harvest index,nitrogen harvest index,nitrogen production efficiency and nitrogen use efficiency of wheat were significantly improved.Under the same nitrogen application rate of N1 and N2,after anthesis weak light stress significantly reduced the accumulation of protein and starch in soft wheat grains,which in turn affected the formation of grain weight,resulting in a decrease in yield.

In order to analyze the alterations in the expression profile of circular RNA(circRNA)in chicken liver cancer cells line(LMH)infected by fowl adenovirus serotype 4(FAdV-4),and the regulatory role of circRNA in the FAdV-4 infecting process,transcriptome sequencing was carried out on FAdV-4-infected LMH cells and uninfected ones.Enrichment analysis of GO functions and KEGG signaling pathways was executed for differentially expressed circRNAs,and five randomly selected circRNAs were verified by Real-time Fluorescent Quantitative PCR(qRT-PCR).The results demonstrated that the circRNAs in the infected and uninfected groups were distributed on the preponderance of chromosomes,and their lengths were mainly concentrated between 300 and 1 000 bp.Differential expression analysis identified 72 circRNAs,with 32 showing significantly upregulated expression levels and 40 presenting downregulated expression levels.GO functional analysis revealed that the genes from which the differential circRNAs originated were mainly enriched in processes such as cellular processes,metabolic processes,catalytic activity,and nucleic acid-binding transcription factor activity.KEGG pathway analysis indicated that the differentially expressed circRNAs were primarily enriched in the Notch signaling pathway,RNA degradation,and the MAPK signaling pathway.The qRT-PCR results showed that the expression levels of the five verified circRNAs were consistent with the sequencing results,further validating the reliability of the sequencing results.This study analyzed the expression profile of circRNAs in FAdV-4-infected LMH cells and screened out differentially expressed circRNAs,providing data support for exploring the functions of circRNAs during the FAdV-4 infection process and the interaction mechanism between the host and FAdV-4.

In order to investigate the effects of different mulching measures on soil moisture and heat status and yield of winter wheat in northwest dry area,winter wheat Kangzhuang 974 was used as test material.From September 2022 to July 2023,three different coverage treatments including wheat straw strip covering three rows(M3),four rows(M4)and five rows(M5)and plastic film covering(PM)treatment were set up in the experimental base of Gansu Agricultural University,The open field(CK)was used as the control.The results showed that:compared with CK,the soil water storage of 0—200 cm during the whole growth period of winter wheat was significantly increased by mulch,the average increase of straw strip mulch was 13.22%,the increase range was M3>M4>M5,and the PM increased by 19.65%.The soil moisture increasing effect of mulch increased gradually with the progress of growth stage,and the maximum increase was 37.53—87.76 mm at maturity stage.It decreased with the deepening of soil layer,and the increase of 0—20 cm was the largest,ranging from 5.10—9.48 mm.Mulching significantly reduced the total water consumption and total water consumption intensity during the whole growth period,and the influence of mulching on water consumption and total water consumption intensity during the later growth period was most obvious.Compared with CK,wheat straw strip mulching significantly reduced the soil temperature of 0—25 cm during the whole growth period by 1.60—2.70 ℃,and M3 treatment had the largest decrease.The maximum decrease occurred at the grouting stage(3.67 ℃),and the maximum decrease between soil layers(3.01 ℃)occurred at 5 cm.Compared with CK,PM significantly increased the soil temperature from 0 to 25 cm during the whole growth period by 1.50 ℃,and the overwintering period and 5 cm had the largest increase,which were 2.20,1.79 ℃,respectively.The temperature of straw mulch increased at 7:00 at wintering stage,jointing stage and ripening stage,and the temperature increased and decreased at other times.PM temperature increased at all time except at 14:00 in grouting stage and ripening stage.Compared with CK, the yield and water use efficiency of M5 and PM were increased by 8.67%, 26.49% and 0.96, 2.94 kg/(ha·mm), panicle number was the most significant factor(CV=17.67%).Yield was significantly positively correlated with spike number(r=0.754^**),WUE(r=0.891^**)and soil temperature(r=0.723^**),and significantly positively correlated with grain number per spike(r=0.522^*).Banded mulching of wheat stalk can achieve both ecological and economic benefits,and M5 is more conducive to the formation of yield.

In order to explore the effects of silicon fertilizer on lodging resistance,growth and development,yield,and quality of winter wheat,Kenmai 58 was used as the experimental material.The treatments included 15 kg/ha (Si1) and 30 kg/ha(Si2)silicon fertilizer applied as basal fertilizer,15 kg/ha (Si3) and 30 kg/ha(Si4) silicon fertilizer applied as topdressing at jointing stage,and no silicon fertilizer (CK).The stem lodging resistance at grain filling stage,plant growth,yield,and quality were measured.The results showed that Si4 significantly reduced the plant height,height of gravity center,and the length of the basal second internode.Compared with CK,Si3 increased the thickness of the basal second and third internodes by over 14.7%.Si2 and Si4 increased the plumpness and breaking resistance of the basal second or third internodes by 8.6%—18.7% and by 12.4%—49.2%,respectively.Nevertheless,silicon fertilizer had no effect on the diameter and dry weight of the basal internode.Moreover,silicon fertilizer increased the chlorophyll content (SPAD) of leaves at jointing stage,but had no effect on tillering,productive tiller percentage,dry matter accumulation,harvest index,yield,and main quality indexes.In summary,the application of silicon fertilizer improved the stem lodging resistance by lowering plant height,gravity center height,and basal internode length,while increasing the basal internode thickness,plumpness,and breaking resistance.Silicon fertilizer also increased chlorophyll content (SPAD) without adversely affecting wheat growth,grain yield,or quality.Silicon fertilizer could be applied either as basal fertilizer or topdressing at jointing stage.

The purpose of this study was to screen the candidate genes and key pathways related to traits at first laying through transcriptomic analysis of liver tissues on laying and non-laying Kangle yellow chickens,and to provide a theoretical basis for studying the molecular mechanisms of laying traits regulation in the liver of chickens.It also provides certain reference for the selection and breeding of Kangle yellow chickens.Three individuals of liver tissues at 154 days of age that had started laying (Group H) and had not started laying (Group L) were selected.Total RNA was extracted using the TRIzol method,and transcriptome sequencing was performed using the Illumina sequencing platform.Differentially expressed genes between the two groups were identified.The differentially expressed genes were subjected to functional enrichment and protein-protein interaction analysis.Nine candidate genes were randomly selected,and their expression levels were verified by qRT-PCR in the livers of the nine individuals.A total of 21 465 genes were detected to be expressed in the liver tissues,and a total of 227 differentially expressed genes were identified,among which 48 were up-regulated and 179 were down-regulated.The qRT-PCR validation results showed that the expression trends of nine genes in the two groups of individuals were basically consistent with the RNA-Seq results,and they showed a decreasing or increasing trend in the H,M(to be laid),and L three groups.Six candidate genes for the age at egg-laying trait were initially identified,namely VTG1,VTG2,VTG3,APOV1,RBP,and RNF186.The five crucial signaling pathways were fat digestion and absorption,cholesterol metabolism,ECM-receptor interaction,estrogen signaling pathway,D-glutamine and D-glutamate metabolism.Six genes and five key signaling pathways were preliminarily identified related to the traits at the first laying of Kangle yellow chicken.

In order to deeply explore the diversity of mycoviruses in Ustilaginoidea virens,this study used an abnormal strain Uv263 of U.virens isolated from diseased rice samples collected from Hainan Province as experimental material to identify potential mycoviruses in this strain,and analyze the relationship between the genome organization and function of mycoviruses.The results showed that strain Uv263 was infected by a novel mycovirus named Ustilaginoidea virens RNA virus 7 (UvRV7).UvRV7 was a double stranded RNA virus with 5 082 bp in total length and 60.29% GC content.UvRV7 encoded two large open reading frames (ORF1 and ORF2),which encoded the coat protein (CP) and RNA-dependent RNA polymerase (RdRP),respectively.The BlastP comparison showed that the RdRP amino acid sequence of UvRV7 shared the highest similarity with that of Thelebolus microsporus totivirus 1,at 48.49%.The results of multiple alignment based on the amino acid sequence of UvRV7 RdRP showed that the RdRP sequence contained a total of eight conserved motifs,among which the most typical GDD motif in the RdRP conserved domain was identified in the Ⅵ motif.The phylogenetic analysis showed that UvRV7 was the most closely related to Thelebolus microsporus totivirus 1 and clustered with representative viruses of the genus Victorivirus in the family Totiviridae.The results of genome organization and evolutionary analyses both indicated that UvRV7 was a novel mycovirus in the genus Victorivirus.Transmission electron microscopy observations showed that UvRV7 formed a spherical viral particle of about 45 nm.Horizontal and vertical transmission experiments showed that UvRV7 could be efficiently transmitted vertically by conidia and efficiently transmitted horizontally between vegetatively compatible strains.Taken together,this study elucidated the genome organization and evolutionary relationships of the novel mycovirus UvRV7 in U.virens,and provided a potential biocontrol agent and theoretical basis for the biological control of rice false smut.

To investigate the effects of delayed sowing on the growth,development,grain yield and quality of wheat,Yunong 907(YN907)and Yunong 922(YN922),two new high yield and high quality wheat varieties,were selected as experimental materials.Three sowing dates were established:October 22nd(S1,the conventional sowing date),October 31st(S2,delayed sowing by 9 days),and November 9th(S3,delayed sowing by 18 days).The study investigated the effects of delayed sowing on the phenological stages,grain yield and flour quality of wheat,and analyzed the relationship between temperature characteristics and wheat yield,and quality under delayed sowing conditions.The results indicated that compared with the conventional sowing date, the full growth period of YN907 and YN922 was shortened with the delay of sowing date in both years. The accumulated temperature before winter and the average daily temperature before heading decreased with the delay of sowing date, while the average daily temperature and effective accumulated temperature after heading increased. The effective accumulated temperature of the full growth period showed a downward trend. The effective accumulated temperature of the full growth period of YN907 and YN922 in S3 treatment was 243.95,222.10 ℃·d lower than that in S1 treatment in 2022—2023, and 136.30,189.40 ℃·d lower in 2023—2024, respectively. The yield and its components decreased with the delay of sowing, with a yield reduction of 6.45% to 17.26%. Compared with the conventional sowing date, the wet gluten content and protein content increased under the delayed sowing conditions.Under the background of climate warming and the scale-up of the agricultural operation,the sowing date of wheat can be adjusted to adapt to the temperature change.YN907 and YN922 had the highest yield on October 22nd.Delaying the sowing date to October 31st could maintain the higher yield level and increase the grain protein content.When the sowing date continues to be postponed,the grain protein content will be significantly increased but the yield will be significantly decreased.

In order to study the effects of different fertilization treatments on the growth and development of aromatic rice,the present experiment was conducted with Qingxiangyou 19 xiang as the experimental material,and five fertilization treatments were designed,broadcasting compound fertilizer(T1),6 cm deep application of compound fertilizer(T2),broadcasting urea(T3),6 cm deep application of urea(T4),and no fertilizer applied treatment(T5),to explore the impacts of different fertilizer applications on the yield,quality,aroma,photosynthetic efficiency,and several other physiological properties of aromatic rice.The results of the experiment showed that the different fertilizer treatments had significant effects on the yield and quality of aromatic rice.The yields of aromatic rice were significantly higher in the deep-fertilization treatments(T2 and T4)than broadcasting-fertilization treatments(T1 and T3).In addition,the yield of aromatic rice was 19.61%,20.03%,39.57% and 32.28% higher than T5 treatment under T2 and T4 treatments in 2022 and 2023,respectively.In terms of net leaf photosynthetic rate,deep fertilization treatments significantly increased the net photosynthetic efficiency of aroma rice leaves by 25.69%,15.95%,17.83% and 11.28% under T2 and T4 treatments in 2022 and 2023,respectively,compared with T5 treatment.Moreover,2-acetyl-1-pyrroline(2-AP)content,2-AP synthesis-related precursor contents,and major enzyme activities were increased in aromatic rice under the deep fertilization treatments.Compared to the T5 treatment,the 2-AP content was significantly increased in the T2 treatments,reaching 161.31,180.17 μg/kg in 2022 and 2023,respectively.Furthermore,a significant increase in precursor content and major enzyme activities were also observed under deep fertilization treatments.The contents of proline,pyrrolidine-5-carboxylic acid and 1-pyrrolidine were increased by 9.90%,10.08%,4.38% and 8.13%,8.26%,6.06% under T2 treatment in 2022 and 2023,respectively.The activities of pyrrolidine-5-carboxylic acid synthetase and proline dehydrogenase activities were enhanced by 8.72%,27.79%,5.52% and 30.91% under the T2 treatment in 2022 and 2023,respectively.In conclusion,the deep fertilization treatment was able to significantly increase the yield,quality,net photosynthetic rate of leaves and promote the biosynthesis of 2-AP in aromatic rice.

IQM(IQ-motif containing protein),a plant-specific calmodulin-binding protein,plays crucial roles in plant growth,development,and responses to various stresses.In order to study the characteristics and potential functions of the maize IQM gene family,bioinformatics approaches were used to identify IQM genes in the maize whole genome,and protein properties,phylogenetic relationship,gene structure,chromosome location,gene replication,cis-acting element,tissue-specific expression and expression patterns under various stresses were investigated.A total of 11 ZmIQMs genes were identified in the whole genome of maize,named ZmIQM1 to ZmIQM11 based on their chromosomal locations.ZmIQMs genes could be classified into three subfamilies,with genes within different subfamilies exhibiting similar structures.Segment duplication was found to play a major role in the amplification and evolution of the ZmIQMs gene family.Cis-acting element analysis showed that the promoter region of ZmIQMs gene contained multiple hormone and stress response elements.The expression pattern of ZmIQMs genes was investigated,and it was found that ZmIQMs genes had different expression patterns in different tissues,and the expression levels of several ZmIQMs genes were changed under different abiotic and biotic stresses.qRT-PCR results showed that under drought stress,the expression of ZmIQM3,ZmIQM4 and ZmIQM10 was up-regulated, and ZmIQM3,ZmIQM4,ZmIQM5,ZmIQM10 and ZmIQM11 responded to Cochliobolus heterostrophus infection.The results showed that ZmIQMs genes played an important role in stress response.

Cloning the wheat grain superoxide dismutase(SOD)gene and developing competitive allele-specific PCR(KASP)markers related to SOD activity are of great significance for breeding wheat varieties with high SOD activity.According to the gene ID,specific primers were designed to clone the gDNA sequence of TaSOD-B1 gene.The single nucleotide polymorphism(SNP)loci were obtained by comparing the wheat genome genetic variation and Ensembl Plants database,and the KASP markers closely related to the SOD activity of wheat were developed.The practicability of the markers was verified by the correlation analysis between SOD activity and genotypes of 287 winter wheat varieties(lines).The TaSOD-B1 gene fragment of Chinese spring variety was amplified by six pairs of specific primers,and the TaSOD-B1 gene on chromosome 5B was obtained by splicing.The full length of the gene was 6 491 bp,including an open reading frame(ORF)of 1 650 bp,which encoded a total of 549 amino acids.The predicted molecular weight was 60.80 ku,the gene was composed of 12 exons and 11 introns.The intron conformed to the typical GT-AG structure.The KASP marker was developed based on the 44th base of the first exon of the TaSOD-B1 gene,and was verified by sequencing.The results showed that the genotype of the allelic variation type TaSOD-B1a was AA,which was associated with high SOD activity,and was labeled with the fluorescent gene FAM(shown as blue).The genotype of TaSOD-B1b was GG,which was associated with low SOD activity and was marked with the fluorescent gene HEX(shown as red).The detection of 287 winter wheat varieties(lines)at home and abroad showed that the SOD activity of different genotypes was significantly different.Based on the TaSOD-B1 sequence,a set of KASP markers related to SOD activity was successfully developed and could be used for genetic improvement of SOD activity.

Based on quantitative trait locus(QTL)mapping of grain weight and grain shape-related traits in a backcross inbred line(BIL)population derived from the cross between indica rice variety Changhui 891 and japonica rice variety 02428,this study provided valuable genetic resources for elucidating the genetic basis of grain development and molecular breeding for high yield and quality in rice.A high-density genetic map was used to conduct QTL analysis for thousand-grain weight,grain length,grain width,grain thickness,and length-width ratio under two environments.Transcriptome data of developing grains at 5,10,and 15 days after flowering from both parents were integrated to analyze candidate genes within QTL regions.Frequency distribution indicated that grain weight and shape traits in the BIL population exhibited typical quantitative characteristics,confirming the suitability for QTL mapping.A total of 37 QTLs were detected across both environments,including 18 specific to Hainan(Sanya),12 specific to Jiangxi(Nanchang),and 7 consistently identified in both locations.These QTLs were distributed on chromosomes 1—5,7,8,11,and 12,comprising 7 for thousand grains weight,5 for grain length,7 for grain width,11 for length-width ratio,and 7 for grain thickness.The analysis showed that the LOD values of these QTLs ranged from 2.79 to 43.10,with a contribution rate of 1.36% to 46.12%.By integrating differentially expressed genes from developing grains of both parents at three stages,28 candidate genes were prioritized from the 37 QTL regions.Among these,LOC_Os11g10480 and LOC_Os11g10100 were identified as key candidates for further functional validation.

To improve the growth,yield,and quality of cantaloupe under extreme high temperature weather in Turpan,Xinjiang,the effects of spraying prohexadione-calcium(PCa)on the physiological growth of cantaloupe under high temperature stress were investigated,distilled water(CK)and PCa with concentrations of 20(PCa1),50(PCa2),100(PCa3),150 mg/L(PCa4)were sprayed on cantaloupe leaves.Through comprehensive analysis of indicators such as photosynthesis,reactive oxygen species content,antioxidant enzymes,antioxidant substances,vine length,stem thickness,yield,and quality of cantaloupe under high temperature stress,the optimal concentration of PCa suitable for foliar spraying of cantaloupe in the region was found.The results showed that as the concentration of PCa increased,the chlorophyll a,chlorophyll b, $\mathrm{O}_{2}^{-}$,H₂O₂,and stem diameter of cantaloupe gradually increased at different stages,with increases of 9.25%-36.29%,4.25%-49.92%,21.45%-334.55%,5.36%-109.41%,and 2.33%-20.69% compared to CK,respectively;and MDA gradually decreased,with a decrease of 7.37%-48.83% compared to CK,respectively.Spraying PCa increased photosynthesis and reactive oxygen species in cantaloupe under high temperature stress,reducing the damage of high temperature to cantaloupe biofilm.PCa1,PCa2,and PCa3 treatments increased the levels of soluble protein,soluble sugar,PRO,SOD,POD,AsA,GSH,yield,soluble solids content,and soluble sugar content of the fruit in cantaloupe under high temperature stress compared to CK.Among the three treatments,PCa2 treatment showed better performance in terms of various indicators.Spraying PCa at an appropriate concentration significantly improved the osmotic regulation substances,antioxidant enzymes,antioxidant substances,and yield and quality of cantaloupe under high temperature stress,enhanced its heat resistance,and achieved increased yield and quality of cantaloupe.Although PCa4 treatment increased the yield of cantaloupe,it reduced the content of soluble solids and soluble sugars in the fruit.High concentrations of PCa delayed the growth of cantaloupe and affected its quality at harvest.Therefore,PCa2 treatment in production is the best treatment to achieve heat resistance,yield increase,and quality improvement of cantaloupe under high temperature stress.It is recommended that the optimal concentration for spraying PCa in this area is 50 mg/L.

To explore the potential biological functions of HBD family members in the important cereal crop wheat,it first conducted a bioinformatic analysis of the HBD family members and their sequence characteristics in common hexaploid wheat.Subsequently,transcriptome and Real-time Quantitative PCR(qRT-PCR)analyses were performed to assess their expression patterns and functions.The results identified a total of 90 wheat HBD genes,which contained between 2 and 18 exons and comprised 111 to 1 863 amino acids;they could be divided into six subgroups based on their evolutionary relationships.The tissue expression pattern results showed that most HBD genes were relatively highly expressed in the roots,stems,spikes,and grains of the plants,while their expression in leaves was relatively low,reflecting the diversity of their biological functions.The promoter regions of these HBD members contain 62 types of cis-acting elements,mainly involved in light and hormone regulatory elements that participated in stress responses.Different members of the HBD gene family responded to various abiotic stresses,including phosphorus,salinity,low temperature,high temperature,drought,and heat-drought synergistic stress.Among these,TaHBD23,TaHBD28,TaHBD67,TaHBD78,and TaHBD85 showed significant differential expression under various stresses,serving as important candidate genes for stress response.In response to biotic stress,the number of HBD family genes responding to Fusarium pseudograminearum and Pseudomonas translucens was fewer than those responding to F.graminearum, suggesting their critical role in the response to F. graminearum resistance.Further research on the transcriptome data from wheat Bobwhite materials infected with F.graminearum and treated with water identified 41 HBD genes with significantly changed expression levels.Among them,10 genes overlapped with the database,and quantitative analysis was consistent with the trends in transcriptome data,indicating that TaHBD28/17,TaHBD67,and TaHBD90/84 negatively responded to Fusarium head blight,while TaHBD39/37,TaHBD45,and TaHBD68/79 positively responded to Fusarium head blight.

In order to study the effects of different nitrogen application rates on rice yield,nitrogen uptake and soil fertility,the effects of different nitrogen application levels on rice yield,nitrogen use efficiency and soil tillage fertility in the Yellow River Irrigation Area were studied from 2021 to 2023 using rice variety Fuyuan 4 as the test material.Compared with other treatments,the average yield of 360 kg/ha was 9.4 t/ha,which was 182.94% higher than that of no nitrogen treatment,and 40.72% and 26.34% higher than that of 210,240 kg/ha.From the perspective of yield components,the increase in yield was mainly due to the increase of grain number per spike and the number of effective spikes,and excessive nitrogen application would cause the decrease of 1000-grain weight.The average of the 3-year results showed that the nitrogen use efficiency was up to 26.93% with 240 kg/ha of nitrogen,and the partial productivity of nitrogen fertilizer decreased with the increase of nitrogen application rate.The highest average value of organic matter was 18.60 g/kg after nitrogen application of 210 kg/ha.Compared with the non-nitrogen application treatment,the soil total nitrogen content of each nitrogen application treatment increased by 7.61%—15.67%,and the soil total nitrogen content of the 360 kg/ha nitrogen application treatment decreased with the increase of the experimental year.The total phosphorus content of soil was gradually reduced in the 360 kg/ha nitrogen application treatment,and the total phosphorus content in other nitrogen application treatments was maintained at about 0.85 g/kg.The total potassium content of nitrogen fertilization treatments gradually decreased with the increase of nitrogen application rate,and the total potassium content of the 360 kg/ha nitrogen application treatment decreased by 23.74%,22.16% and 8.85%,respectively,compared with the nitrogen fertilization treatments of 120,210 and 240 kg/ha.Soil available phosphorus increased with the years of the experiment,and the variation range between treatments increased with time.With the increase of nitrogen application rate,soil available phosphorus increased first and then decreased,and the soil available phosphorus of the 240 kg/ha treatment increased significantly in 2023 compared with other treatments.The soil available potassium of each treatment did not change significantly with the test years.By considering crop yield,nitrogen use efficiency and soil fertility,nitrogen application of 240 kg/ha could balance the environment and production requirements.

STAT proteins are a class of transcription factors that play crucial roles in signal transduction and gene transcriptional activation.In plants,the expression of STAT genes are associated with abiotic stresses such as high temperature.To investigate whether maize STAT genes are involved in the response to high-temperature stress,two maize inbred lines,Zheng 58(tolerant to high-temperature stress)and PH6WC(sensitive to high temperature),were selected as materials.The plant tissues from five parts(root,stem,leaf,pollen,and filament)of plants grown under high-temperature and normal-temperature conditions were used for transcriptome sequencing.Based on the sequencing data,the structure of STAT genes,the physicochemical properties of the proteins encoded by STAT genes,and tissue-specific expression patterns of STAT genes under different temperatures and materials were analyzed.The results showed that two STAT genes were identified in maize,named Zm-STAT1 and Zm-STAT2.The protein encoded by Zm-STAT1 was hydrophobic,while that encoded by Zm-STAT2 was hydrophilic,both containing multiple functional and phosphorylation modification sites.Further expression analysis revealed that, with room temperature as the control, under high-temperature conditions, Zm-STAT1 gene was upregulated in the root of PH6WC and in the pollen and filament of Zheng 58, whereas Zm-STAT1 gene in the stem and leaf of PH6WC and Zm-STAT2 gene in the leaf of PH6WC were down-regulated expression. Under both temperature conditions,the expression level of Zm-STAT2 was significantly higher than that of Zm-STAT1 across all five tissues.Notably,Zm-STAT2 was induced by high temperature in root,stem,pollen,and filament in the heat-tolerant Zheng 58,suggesting that Zm-STAT2 gene was involved in high-temperature stress response.

β-Amylase (BAM) is involved in regulating various biological processes in plants and responds to multiple external stimuli such as hormones and abiotic stress. To explore the characteristics and expression patterns of the BAM gene family in maize, we performed a genome-wide identification of the maize BAM gene family using bioinformatics methods. We also analyzed the encoded proteins, chromosomal localization, gene structure,cis-acting elements, synteny analysis, tissue-specific expression, and the expression patterns of the genes after simulated hail stress and melatonin treatment. The results showed that 16 ZmBAM members were identified in maize, all of which contained the typical Glyco_hydro_14 domain, with most of them being hydrophilic proteins. Phylogenetic analysis indicated that ZmBAM protein can be divided into three groups, with genes in the same group sharing similar gene structures and motif distributions. Cis-acting element analysis suggested that the expression of most ZmBAM genes might be related to plant hormones, abiotic stress, and light responses. Synteny analysis revealed a certain level of homology between the maize BAM gene family and those of Arabidopsis and soybean, with a closer relationship to rice. qRT-PCR analysis showed that, under simulated hail stress, most genes were upregulated, and their expression was significantly upregulated after melatonin treatment. This study provided a reference for further understanding the function of the maize BAM gene family in response to abiotic stress.

The effects of different nitrogen reduction and efficiency enhancement measures on ammonia volatilization and crop yield in fluvo-aquic soil of wheat-maize rotation system were studied to provide guidance for rational fertilization and agricultural environmental protection.A long-term nitrogen reduction was carried out since 2016 at the experimental station of Xuchang fluvo-aquic soil area in Henan Province.No nitrogen fertilizer control(CK),conventional nitrogen fertilizer(100N),20% nitrogen reduction(80N),and 20% nitrogen reduction combined with straw returning(80NS),nitrification inhibitor(80NI),and biochar(80NB)were set up.The soil physical and chemical properties,annual ammonia volatilization characteristics and wheat and maize yield of different treatments were studied from 2021 to 2022.In the wheat season,the pH value of 80NS,80NI and 80NB treatments was significantly higher than that of 100N treatment.The organic matter content significantly increased, while the soil bulk density significantly decreased in the 80NS and 80NB treatments, compared with the 80N and CK treatments, respectively. At the base fertilizer stage of wheat,the ammonia volatilization accumulation of 80NS,80NI and 80NB treatments was significantly lower than that of 100N by 28.71%,35.61% and 22.99%,respectively.During the topdressing stage of wheat season,the ammonia volatilization accumulation of 80NS and 80NB treatments was significantly lower than that of 100N by 14.94% and 17.58%,respectively.The ammonia volatilization accumulation of 80NS and 80NI treatments was significantly increased by 22.27% and 27.69% compared with 80N.During the whole growth period of wheat,the accumulation of ammonia volatilization in different nitrogen treatments accounted for 1.31%-2.47% of the nitrogen application rate,showing 100N> 80NB> 80NS> 80NI> 80N.In the maize season,compared with the accumulation of ammonia volatilization under 100N treatment,80N and 80NS treatments significantly decreased by 37.14% and 29.63%,respectively,and 80NI treatment significantly increased by 60.83%.Compared with 80N treatment,the accumulation of ammonia volatilization in 80NI and 80NB treatments increased significantly by 155.79% and 44.05%.The accumulation of ammonia volatilization in maize growth period accounted for 5.81%-14.86% of nitrogen application rate,showing 80NI> 100N> 80NB> 80NS> 80N.The wheat yield results indicated that compared with 100N treatment,80N treatment significantly reduced the yield by 16.67%,while 80NS,80NI and 80NB treatments did not significantly reduce the yield.Maize yield data showed that there was no significant difference between 100N treatment and four nitrogen reduction treatments.In summary,the application of nitrification inhibitors,straw and biochar on the basis of reducing nitrogen by 20% in the experimental fluvo-aquic soil can effectively improve soil fertility and stabilize crop yield.However,nitrification inhibitors and biochar significantly increase the accumulation of ammonia volatilization in maize season,which needs special attention in actual production.

To systematically study genes related to fruit hardness,genome resequencing BSA method was used to locate the fruit hardness association interval,and predict candidate genes based on their corresponding reference genome collinearity segments and gene annotation information,to lay the foundation for the next step of gene localization and cloning.The fruit hardness separation of F₂ offspring from stable genetic soft flesh inbred line C18 and hard flesh inbred line LE4 crosses followed a normal distribution.30 soft fleshed and 30 hard fleshed individual plants were selected from the F₂ population to construct extreme mixed pools,and whole genome resequencing with 30×and 10×coverage on the mixed pools and parents was conducted.A total of 1 891 040 single nucleotide polymorphisms(SNPs)and 376 603 insertion deletion markers(InDels)were obtained from the hybrid pools and parents,which were used for genome-wide mapping of fruit hardness traits.The peak of BSA localization was distributed within a total of 2.71 Mb between 72 610 411 and 75 329 951 bp on eggplant chromosome 6.Based on pathway enrichment and gene function annotation,candidate genes Smechr0601726.1 and Smechr0601735.1 were obtained.In summary,through genome resequencing BSA analysis,eggplant fruit hardness may be regulated by two important candidate genes.Smechr0601726.1 encodes a polygalacturonase gene,which is directly related to fruit hardness;Smechr0601735.1 is closely related to the metabolism of ascorbic acid and arabic acid,encoding ascorbate peroxidase,and is associated with fruit development and hardness formation,which can delay fruit softening.

The effects of tillage depth and organic fertilizer rates on photosynthetic characteristics,yield formation,and economic benefits of wheat were explored to provide a theoretical and technical basis for fluvo-aquic soil or similar soil types.The two-factor split-plot field experiment was conducted in the typical fluvo-aquic soil area of Qihe County,Dezhou City,Shandong Province from 2022 to 2024,where the tillage depth 15-20 cm(D1)and tillage depth 30-35 cm(D2)were assigned to the main plots,and the organic fertilizer rates of 800(L),1 200(M),and 1 600 kg/ha(H)were assigned to the subplots.The photosynthetic characteristics,aboveground dry matter accumulation characteristics and yield composition,in wheat were tested in fluvo-aquic soil under different tillage depths and organic fertilizer rates.Both D2M and D2H treatments were beneficial to improve the yield and yield components of wheat,and the spike number,kernels per spike,1000-grain weight,and grain yield significantly increased by 5.5%-8.5%,3.5%-12.1%,6.7%-13.2% and 6.6%-12.8%,compared with other treatments,respectively.D2M and D2H treatments also stabilized or increased the above-ground dry matter accumulation at jointing,anthesis,and maturity stages by improving the above-ground dry matter accumulation rate at each growth stage,which was 9.0%-22.1%,8.9%-25.8% and 10.7%-24.3% higher than averaged across the two years of other treatments,respectively.Compared with D1,D2 was more conducive to promoting the effect of organic fertilizer on the SPAD of leaves at different growth stages.Under D2M and D2H treatments,higher chlorophyll content could be maintained from the mid- to late-filling stages.Under M and H organic fertilizer application rates,the Pn of leaves at different growth stages of D2 was significantly higher than that of D1.At jointing,booting,flowering,mid-filling,and late-filling stages,on average over the two years,D2M and D2H increased significantly by 12.0% to 16.7%,13.7% to 16.8%,13.8% to 19.7%,20.2% to 25.8%,and 24.6% to 44.8%,respectively,compared with D1M and D1H.Under the same organic fertilizer application rates,the difference in leaf LAI between the two tillage depths gradually increased with the progression of the growth process.D2M and D2H performed best at anthesis and mid-filling stages,increasing by 13.2% to 27.2%,and 13.4% to 29.4%,respectively,compared with other treatments on average over the two years.In conclusion,both D2M and D2H treatments could enhance the photosynthetic characteristics of plants and the above-ground dry matter accumulation ability,thereby optimizing the yield components and achieving the drastically improvement of wheat yield.However,there were no significant differences in the indices between D2M and D2H treatments.Therefore,considering resource conservation,this study suggests that the combination of a tillage depth of 30-35 cm and an organic fertilizer application rate of 1 200 kg/ha can achieve wheat high yielding.

The aim of this study was to investigate the relationship between calcium uptake,indoleacetic acid(IAA)and organic acid metabolism during the development and ripening of Cerasus humilis fruits,in order to provide a basis for further research and applications of Cerasus humilis.The high calcium and low calcium Cerasus humilis fruits from Inner Mongolia were taken as the experimental materials.Samples were taken at different stages of fruit development and ripening,including the young fruit stage(S1),hard kernel stage(S2),coloring and enlargement stage(S3),hard ripening stage(S4),and fully ripe stage(S5),this study investigated the changes of calcium uptake capacity,IAA,and organic acid metabolism,and carried out correlation analysis.The results indicated that the uptake capacity of water-soluble calcium continuously increased throughout the development and ripening of Cerasus humilis fruits,with a marked rise in uptake activity,uptake rate,and uptake amount at the fully ripe stage;the uptake capacity of calcium pectin was higher from the young fruit stage to hard kernel stage but declined in the later stages of fruit development,with a significant reduction in uptake activity,uptake rate,and uptake amount at the fully ripe stage;the uptake capacity for active calcium and total calcium followed trends similar to those observed for calcium pectin.During fruit development and ripening,the IAA content in Cerasus humilis fruits showed a pattern of initially increasing and then decreasing,with significantly higher levels during the the young fruit stage and hard kernel stage compared to other stages.The activity of NAD-MDH,as well as malic acid content and organic acid total content,generally increased,peaking at the hard ripening stage.The activity of NADP-ME declined overall,while citric acid content increased initially and then decreased,reaching its highest point at the hard kernel stage.Correlation analysis revealed varying degrees of correlation between IAA content,organic acid metabolism indicators,and the uptake capacity of different calcium forms in high-calcium and low-calcium Cerasus humilis fruits.Specifically,IAA content was positively correlated with the uptake capacity of total calcium;the activity of NAD-MDH,malic acid content,and organic acid total content was negatively correlated with total calcium uptake capacity;while the activity of NADP-ME and citric acid content was positively correlated with total calcium uptake capacity.During the development and ripening of Cerasus humilis fruits,calcium uptake is associated with IAA content and organic acid metabolism.High levels of IAA and citric acid contents during the early stages of fruit development significantly promote calcium uptake,whereas enhanced malic acid synthesis and metabolism in the later stages of fruit development inhibits calcium uptake.

In order to explore the effect of partial replacement of chemical fertilizer by organic fertilizer on the ecological stoichiometric characteristics of soil carbon,nitrogen and phosphorus in the Loess Plateau of Eastern Qinghai,a field experiment was conducted with continuous cropping of potato(Qingshu 9).This field experiment was established in 2022 with five treatments,i.e.no fertilizer(CK),100% chemical fertilizer(T1),30% organic fertilizer+70% chemical fertilizer(T2),50% organic fertilizer+50% chemical fertilizer(T3),and 70% organic fertilizer+30% chemical fertilizer(T4).Soil contents of organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus,microbial biomass carbon,nitrogen,phosphorus and soil-microbial biomass ecological stoichiometry were determined.The results showed that compared with 2022,the contents of soil organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus in each fertilization treatment in 2023 showed an increasing trend,while the CK treatment showed a decreasing trend.Among different soil layers,the content of each index gradually decreased with the increase of soil depth.Among different treatments,according to the content of each index,the order was T3>T4>T2>T1>CK.In the 0—30 cm soil layer,the contents of soil organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus under T3 treatment were 6.88%,17.65%,17.88%,84.71%,77.01%,65.67%,80.07%,54.91% higher than those of CK,respectively.The soil C∶N and C∶P in each soil layer were the highest in the CK treatment.Among the four fertilization treatments,soil C∶N,microbial biomass C∶N and C∶P were the lowest in the T3 treatment.In addition to the 0—10 cm soil layer in 2023,the soil C∶P in each soil layer was also the lowest in T3,while the microbial biomass N∶P in the T1 treatment was the lowest among all fertilization treatments.Soil C∶N and C∶P increased with soil depth.Correlation analysis showed that organic carbon,total nitrogen,total phosphorus,available nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus were significantly positively correlated with each other.In summary,the substitution of organic fertilizer for chemical fertilizer can not only change the soil nutrient and soil microbial biomass content,but also change the soil ecological stoichiometry characteristics,and the treatment with 50% organic fertilizer+50% chemical fertilizer has the best effect.

Wheat,as one of the world's major food crops,its growth and development,yield,and quality are often constrained by abiotic stresses. To cope with these abiotic stresses,wheat has evolved a series of response mechanisms,including signal perception,signal transduction,and gene expression regulation,to maintain cellular homeostasis and protect the functions of biological macromolecules. Drought stress causes the accumulation of osmotic-adjustment substances and antioxidants in wheat,a decline in the photosynthetic rate,and alterations in morphological structure. Its molecular mechanisms mainly involve pathways such as reactive oxygen species (ROS) signaling,hormone signaling,and gene expression regulation. Saline-alkali stress inhibits seed germination and seedling growth,leading to photosynthetic impairments,osmotic-regulation imbalances,ion-balance disorders,and membrane-lipid oxidation. Under such circumstances,wheat can enhance salt tolerance by activating the salt overly sensitive (SOS) signaling pathway,calcium-ion signaling,hormone signaling,and gene-expression regulation. Extreme temperature stress inhibits photosynthesis,triggers membrane-lipid oxidation,and promotes the accumulation of osmotic-adjustment substances. Its regulatory mechanisms mainly involve hormone signaling,transcriptional activation,and epigenetic regulation. This study reviews the response mechanisms of wheat to drought,saline-alkali,and extreme temperature stresses and looks ahead to future research directions:focusing on the cross-interaction mechanisms of multiple stresses and mining genes responsive to combined stresses; effectively integrating stress-resistant traits into high-yield and high-quality genetic backgrounds to achieve the coordinated improvement of ideal plant types and stress resistance; promoting the in-depth integration of modern biotechnology with conventional breeding methods to enhance breeding efficiency. It aims to provide comprehensive theoretical support and technical references for the genetic improvement of wheat's resistance to abiotic stresses.

As an important subfamily of receptor kinases,cysteine-rich receptor kinases play a key role in plant growth and development.The aim of this study was to analyze the relationship between the GhCRK26 gene and the development of cotton fiber,and to provide a theoretical basis for the improvement of cotton fiber quality.This study selected upland cotton Line 9 and sea island cotton Xinhai 16,and collected samples at different periods of fiber development and root,stem and leaf tissues.The expression pattern of GhCRK26 was analyzed by qRT-PCR;the gene was cloned by PCR;a phylogenetic tree was constructed with the help of bioinformatics tools to predict the physicochemical properties of the protein,its transmembrane structure and signal peptide;the promoter cis-acting elements were analyzed by the PlantCare database;and the localization of the protein was clarified by the subcellular localization assay.The results showed that the expression of GhCRK26 gene showed highly significant differences at 10,20 and 30 days after flowering in two varieties;the highest expression levels were found in the leaves of Line 9,while in Xinhai 16,the expression of stem and leaves did not show significant differences.A 2 049 bp CDS sequence encoding 682 amino acids was successfully cloned.The evolutionary tree showed that GhCRK26 protein was the most distantly related to Hibiscus trionum,and the closest to Gossypium tomentosum and Gossypium barbadense;the protein was hydrophilic and unstable,containing a transmembrane structure and a signal peptide;methyl jasmonate and abscisic acid response elements were identified in the promoter region;and the subcellular localization confirmed that it was localized in the cell membrane.The above studies provide a basis for further in-depth analysis of the function of GhCRK26 gene in fiber development.

The TCP transcription factor family is unique to plants and plays a crucial role in key biological processes such as plant growth,metabolic activities,and stress responses.To investigate the quantity,distribution and expression patterns of TCP gene family in the Amorphophallus konjac genome,bioinformatics approaches were utilized to identify the AkTCP gene family,followed by a comprehensive analysis of the physicochemical properties,subcellular localization,collinearity,gene structure,and evolutionary relationships.Additionally,gene expression in response to biotic and abiotic stresses was examined by qRT-PCR.The findings were summarized as follows:the A.konjac genome contained 30 AkTCP family genes,with protein lengths ranging from 135 to 562 amino acids.These proteins had molecular weights between 15.08 and 57.20 ku,and isoelectric points between 4.96 and 11.49.Most of these were basic,hydrophilic,and unstable proteins,predominantly localized in the nucleus.These genes were unevenly distributed across the chromosomes.The collinear AkTCP genes were unevenly distributed across eight chromosomes,comprising four inter-chromosomal and five intrachromosomal collinearity events.The AkTCP gene structure was relatively simple,with most genes lacking introns,and all AkTCP transcription factors exhibit a highly conserved TCP domain.Phylogenetic classification separates the 30 AkTCP genes into two major classes,Class Ⅰ and Class Ⅱ,with Class Ⅱ further subdivided into the CIN and CYC/TB1 subclasses.The promoters of AkTCP genes contain cis-elements related to physiological response,light-responsive,phytohormone-responsive and stress-responsive elements.The results of qRT-PCR analysis indicated that AkTCP family members were involved in and responded positively to drought stress, salt stress, methyl jasmonate and soft rot pathogens of A. konjac.

This study aimed to investigate the impacts of three exogenous amino acids on the growth and development of tomato seedlings under high-temperature stress.Tomato seedlings were selected as the study material.The normal temperature treatment with clean water at 25 ℃/18 ℃(day/night)and the high-temperature treatment with clean water at 40 ℃/30 ℃(day/night)served as control and heat groups,respectively.Under high-temperature,tomato seedlings were sprayed with 5 mmol/L γ-aminobutyric acid(GABA),0.2 mmol/L 5-aminolevulinic acid(ALA),and 5 mmol/L Proline(Pro)on the leaves once every seven days for two times.Growth parameters,antioxidant enzyme activities,and physiological parameters were determined.The results indicated that the plant height,stem diameter,and biomass accumulation of tomato seedlings was significantly reduced under high temperature.Compared with heat treatment,spraying GABA,ALA,and Pro significantly enhanced the plant height and stem thickness of tomato seedlings,promoted root development,increased biomass accumulation and relative growth rate under high-temperature.Moreover,the chlorophyll content and net photosynthetic rate of tomato seedlings declined remarkably.The chlorophyll content in the ALA treatment was higher than that in other treatments,while the net photosynthetic rate in the GABA treatment was the optimal.Compared with heat treatment,spraying GABA and ALA significantly augmented the activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT).High-temperature stress substantially elevated the relative electrical conductivity(REC)and malondialdehyde(MDA)content of leaves.Meanwhile,the leaves treated with Pro exhibited the lowest REC and MDA content,nearly returning to the level of control treatment.The outcomes of cluster and principal component analyses revealed that the GABA and ALA treatments mainly enhanced the photosynthetic rate and promoted plant growth by coordinately regulating the chlorophyll content and antioxidant enzyme activities of tomato seedlings,thereby strengthening the capacity to resist high-temperature stress.In contrast,Pro treatment effectively regulated the leaf's osmotic pressure by reducing leaf relative conductivity and MDA content.

To investigate the role of the BnMLP2 gene in drought tolerance in ramie,the BnMLP2 gene encoding a metallothionein-like protein in ramie was obtained by analyzing ramie transcriptome data using Zhongzhu-1 as the material.The BnMLP2 gene was screened and cloned from the ramie transcriptome data,and bioinformatics analyses were conducted,including sequence alignment,domain prediction,and subcellular localization prediction.Fluorescent Quantitative PCR was used to determine the expression profile of the BnMLP2 gene in different tissues of ramie and to explore its expression changes under drought stress.The BnMLP2 gene was introduced into Arabidopsis thaliana to construct transgenic plants.Under drought stress conditions,phenotypic,physiological,and biochemical differences between transgenic and wild-type Arabidopsis were compared,along with the expression of stress-related genes.The results showed that the open reading frame of the BnMLP2 gene in ramie was 243 bp in length,encoding 80 amino acids.BnMLP2 had the closest amino acid sequence homology to metallothionein(MT)or metallothionein-like protein (MLP) from Malus domestica,both belonging to the metallothionein family; it contained the PFAM01439 domain and belonged to class Ⅱ metallothionein,with a predicted subcellular localization in the cytoplasm.The BnMLP2 gene was expressed in all tissues of ramie,and its expression was significantly induced by drought,especially in stems.Under drought stress,transgenic Arabidopsis overexpressing BnMLP2 exhibited stronger drought tolerance compared to wild-type plants,as evidenced by significantly increased root length and fresh weight,enhanced antioxidant enzyme and γ-glutamylcysteine ligase (γ-GCL) activities,and accumulation of more proline (Pro),glutathione(GSH),glutathione disulfide (GSSG),and phytochelatins (PCs) to regulate intracellular ion homeostasis.The contents of malondialdehyde (MDA)and hydrogen peroxide (H₂O₂)in transgenic lines were significantly lower than in wild-type plants,at 55% and 80% of wild-type levels,respectively.In addition, the content of sodium and potassium ions in transgenic Arabidopsis under drought conditions was 4.4 times and 1.4 times higher than that of the wild type, respectively. Overexpression of BnMLP2 induced increased expression of three stress-related genes,AtMT1a,AtNCED3,and AtWRKY40,with maximum expression levels of 1.5,1.9,and 2.8 times those in wild-type plants,respectively.These results suggest that the BnMLP2 gene enhances the tolerance of Arabidopsis to drought stress.

This study aims to identify potential proteins that interact with the coat protein(CP)of Tomato chlorosis virus (ToCV)from a cDNA library,to explore the infection mechanism of ToCV and the role of CP in the infection process.The research highlights the significant impact of Tomato chlorosis virus disease on tomato yield and quality during summer and late-autumn production.The Moneymaker tomato variety infected with ToCV was used as the experimental material.Using Gateway technology,a nuclear yeast cDNA library was constructed from ToCV-infected tomatoes,and a yeast two-hybrid bait vector,pGBKT7-CP(CP-BD),was developed.CP was employed as the bait protein to screen the nuclear yeast cDNA library,identifying hundreds of potential interacting proteins involved in various physiological processes.Further verification was performed using one-on-one yeast two-hybrid assays and NCBI BLAST analysis to confirm the proteins interacting with ToCV CP.The constructed nuclear yeast cDNA library had a primary capacity of 1.60×10⁷,with a 100% recombination rate and an average insert size exceeding 1 000 bp.The secondary library also achieved a capacity of 1.60×10⁷,with a 100% recombination rate and an average insert size greater than 1 000 bp,meeting the quality standards for subsequent yeast hybridization experiments.Proteins interacting with ToCV CP,identified through library screening,were categorized into cellular processes,biological regulation,and intracellular material transport.Notably,many of these proteins were associated with processes such as viral replication and transport,host cell infection,and the regulation of host cell metabolism and the cell cycle.Additionally,the identified proteins included those with functions such as protein binding,nucleic acid binding,and hydrolase activity.Among these,ribonucleases were the most abundant,playing a critical role in the viral infection process.Ultimately,30 proteins,including HSPs,DnaJ,and TCPs,were confirmed to interact with ToCV CP.These findings provide a strong foundation for further research into the infection mechanism of ToCV and the functional role of CP in the infection process.

In order to explore the role of NBS-LRR gene RPM1 in the resistance to Verticillium wilt in Solanum,it took the wild Solanum sisymbriifolium Lam.as material,and cloned the homologous sequence of RPM1 gene on the basis of its transcriptional sequencing.The physicochemical properties and molecular structure of the sequence encoded protein were analyzed,the evolutionary relationship tree was constructed,and subcellular localization was performed in tobacco.At the same time,the relative expression level of RPM1 gene in different parts of Solanum sisymbriifolium Lam.was detected,as well as the relative expression level at for time points(0,24,48 and 72 h)after inoculation with Verticillium dahliae(a pathogen of Verticillium wilt).The results showed that the total length of RPM1 gene(SsRPM1)was 2 772 bp,encoding 924 amino acids.SsRPM1 protein,with a total molecular weight of 105.99 ku,was an alkaline hydrophilic protein without transmembrane structure.SsRPM1 protein was mainly composed of α helix and random coil,including LRR,NBC and CC domains.Solanum dulcamara RPM1 protein had the closest relationship with it.Subcellular localization in tobacco found that the protein was located on the cell membrane.SsRPM1 gene was expressed in different organs(root,stem and leaf),among which the stem had the highest relative expression,followed by leaf and root.After inoculation with V.dahliae,in general,SsRPM1 gene expression in both control group and inoculation group showed a trend of first increasing and then decreasing.The relative expression level of SsRPM1 gene was the highest at 24 h after inoculation.Compared with the control group,the relative expression level of SsRPM1 gene in inoculation treatment was lower.It is suggested that SsRPM1 is a negative regulatory gene in response to Verticillium wilt stress.

OsAAP8 overexpressed transgenic lines were constructed based on the japonica rice variety Zhonghua 11.By detecting and analyzing its agronomic and quality traits,it explored the effects of OsAAP8 overexpression on rice growth and development,as well as rice quality,to provide theoretical basis for molecular design breeding by utilizing OsAAP8 gene to improve rice quality and yield.The results showed that compared with the transgenic negative plants,OsAAP8 overexpressed transgenic positive plants had significantly reduced plant height,tiller number,and single plant yield.The quality trait testing results showed that the content of glutamic acid,threonine,essential amino acids,total amino acids,protein,amylose,gel consistency,and gelatinization temperature in OsAAP8 overexpressed transgenic positive rice were significantly increased.The content of total starch and brown rice rate did not change significantly.The content of free fatty acids,taste value,polished rice rate,and whole polished rice rate were significantly decreased.The observation results of optical microscope and scanning electron microscope showed that the chalkiness rate and chalkiness degree of OsAAP8 overexpressed transgenic positive rice were significantly increased,but the chalkiness area did not change significantly,and the shape and arrangement of starch granules did not change.The grain size detection results showed no significant changes in grain length,width,and thickness of OsAAP8 overexpressed transgenic positive rice.The above results indicate that overexpression of OsAAP8 is not conducive to the growth and development of rice,but can significantly improve the nutritional quality of rice,which provides important information for the cultivation of new high-quality rice varieties.

To investigate the genetic evolution and molecular differences of Lentinula edodes germplasm resources and their hybrid progeny,it utilized whole-genome resequencing technology and analyzed genetic diversity and population structure based on single-nucleotide polymorphism(SNP)data.It involved eight main cultivated varieties,six hybrid new varieties,and two wild strains of L.edodes.The results revealed that the SNP numbers in the wild strains Y5 and Y6 were significantly higher than those in the cultivated varieties,with 158 659 and 149 422 SNPs,respectively,and they clustered in independent branches based on genetic distance.In contrast,the cultivated varieties and their hybrid progeny exhibited similar SNP numbers and genetic relationships,with SNP counts ranging from 97 295 to 105 627 and genetic distances ranging from 0.001 2 to 0.055 3,clustering within the same branch.The branches of the cultivated varieties and their hybrid progeny could be divided into six closely related groups.These included early-maturing varieties L18 and 868,early-maturing white-faced varieties RX11,QK212,and 0912,medium-maturing firm-textured varieties 808,LX1,and 168,hybrid progeny varieties JX15 and JX3 of 808,L18,and 868,hybrid progeny varieties JXB5 and JXB15 of JX15 and JX3,and hybrid progeny varieties E14 and E36 of 0912 and JXB5.Population structure and principal component analysis revealed that L.edodes germplasm resources could be divided into four subgroups.The first class included varieties RX11,QK212,and 0912,as well as their hybrid progeny E14 and E36.The second class consisted of main cultivated varieties LX1,168,and 808,along with their hybrid derivatives JX15 and JX3,which share extremely similar genetic information or background.The third class comprised L18 and 868,along with their multiple-generation,genetically improved,high-quality hybrid strains,JXB5 and JXB15,which exist in a transitional state between cultivated varieties and wild strains,with a propensity towards the cultivated varieties.The fourth class was represented by wild strains YX5 and YX6,which are distinctly different from the cultivated strains.This study clarified the genetic differences among L.edodes strains,laying the foundation for the selection of parents and the pairing of hybrid combinations in subsequent hybrid breeding.

Explore the effects of amino acid type water-soluble fertilizer on chlorophyll content and proportion of celery,and the regulation of gene expression related to chlorophyll metabolism,which provides a theoretical basis for the rational application of amino acid type water-soluble fertilizer foliar fertilizer in the production of high-quality pigment-rich celery.The celery variety Ningqin 1 was sprayed with different concentrations of amino acid type water-soluble fertilizer,the contents of chlorophyll a,chlorophyll b and total chlorophyll in leaf blades and petioles were determined and analyzed,and the relative expression levels of genes related to chlorophyll metabolism were determined by fluorescence quantitative PCR(RT-qPCR).The effects of amino acid type water-soluble fertilizer on chlorophyll accumulation and relative expression levels of related genes were related to the treatment concentration and leaf location.Treatment with 500 μL/L amino acid type water-soluble fertilizer increased the contents of chlorophyll a,chlorophyll b and total chlorophyll in leaf blades and petioles,and decreased the ratio of chlorophyll a/b in petioles.The expression of chlorophyll synthesis-related genes(AgHEMA,AgHEMB,AgCHLM,AgPOR and AgCAO)was induced,and the expression levels of degradation related genes(AgPAO and AgPPH)were significantly down-regulated.Treatment with 1 000 μL/L amino acid type water-soluble fertilizer inhibited chlorophyll accumulation to a certain extent,increased chlorophyll a/b ratio in leaf blades,and significantly down-regulated the expression levels of genes related to chlorophyll synthesis and chlorophyll degradation AgPPH.The suitable concentration of amino acid type water-soluble fertilizer foliar fertilizer can improve the chlorophyll content of celery at seeding stage by regulating the expression level of genes related to chlorophyll metabolism.

To elucidate the distribution characteristics and evolution patterns of total nitrogen(TN)and hydrolytic nitrogen(HN)in irrigated desert soil profiles under long-term fertilization regimes,and to explore effective strategies for enhancing soil fertility and rational fertilization,it utilized soil samples(0-200 cm depth)collected in 2003 and 2022 from a long-term fertilization experiment established in 1982.TN and HN contents were measured across soil profiles,and their dynamic changes in the topsoil after 1,10,20,30,and 40 a of different fertilization treatments were analyzed.The results indicated that TN and HN contents gradually decreased with soil depth increasing under all fertilization regimes.Long-term combined application of organic manure and chemical fertilizers significantly increased TN content in surface soil.At the same time,treatments incorporating organic manure exhibited notably higher HN content in surface soil compared to those without organic inputs.Prolonged sole nitrogen fertilization led to TN profiles similar to those observed in unfertilized control plots.A significant positive correlation was identified between HN and TN across all fertilization regimes.Specifically,every 1.00 g/kg increase in TN under organic-amended treatments corresponded to a 71.57 mg/kg increase in HN.The findings demonstrate that long-term integrated organic-inorganic fertilization significantly enhances both TN and HN contents,thereby improving soil nitrogen fertility.Conversely,nitrogen depletion or excessive nitrogen-only applications reduced soil nitrogen fertility.These results highlight that combined organic and inorganic fertilization represents an effective approach for soil fertility improvement in irrigated desert soil regions.

The purport was to identify the black rot pathogen of bitter gourd (Momordica charantia L.) and screen highly effective biocontrol Bacillus spp.,so as to provide a theoretical basis for scientific prevention and control of black rot of bitter gourd.The pathogen of black rot was isolated and purified by tissue separation method from postharvest bitter gourd,and the pathogenicity of isolated pathogens was further verified according to Koch's rule.The pathogen was identified by observing morphological characteristics combined with molecular phylogenetic analysis.The representative strain NCKG8.2-4 was used as the test pathogen,and antagonistic experiments were conducted to compare the effects of Bacillus subtilis subsp. spizizenii TEB-1,Bacillus amyloliquefaciens SWB-2,Bacillus licheniformis SWB-1,Bacillus velezensis SB023 and Bacillus polymyxa SWP-1 on black rot pathogen of bitter gourd.The inhibition effect of the best antagonistic strains on test pathogen was further evaluated by the gradient inhibition experiment of the cell-free supernatant (CFS) from its fermentation broth.The results showed that NCKG8.2-4 strain was pathogen causing black rot of bitter gourd,and the ITS,β-Tubulin and GAPDH fragments sequences were 100% consistent with Stagonosporopsis cucurbitacearum.And it was respectively clustered with S.cucurbitacearum on a branch by phylogenetic analysis based on β-Tubulin or GAPDH sequences.The results of antagonistic experiment showed that the inhibitory area of SB023 against colony growth of black rot pathogen NCKG8.2-4 was the largest.At the same time,the inhibitory effect increased with the Mcrease of SB023 CFS content,and the inhibitory rate of 9% CFS for NCKG8.2-4 growth was up to 93.7% and the EC₅₀ of SB023 CFS was 3.48% to NCKG8.2-4 on PDA.The pathogen of black rot of bitter gourd was identified as S.cucurbitacearum,and Bacillus velezensis SB023 had a high inhibitory effect on the pathogen NCKG8.2-4.

The screening of aromatic rice germplasm resources and the rapid identification and utilization of aromatic genes have practical significance for the genetic breeding of aromatic rice.In order to clarify the aroma types of aromatic rice materials in Heilongjiang Province and improve the breeding efficiency of aromatic rice varieties,180 aromatic rice varieties approved and promoted in Heilongjiang Province were sequenced.Compared with the reference genome Nipponbare,the sequencing results were divided into two types of mutations.The first type was the 8 bp deletion of exon 7 and three base substitutions(Badh2-E7 type),including 177 varieties.The second type was the 7 bp deletion of exon 2(Badh2-E2 type),which contained only three varieties.According to these two mutation types,two specific KASP molecular markers were designed to accurately detect the aroma types of E7 and E2 in rice.Through the identification of 180 aromatic rice varieties,the typing results were consistent with the sequencing results.Two hybrid populations were constructed by using E7 aromatic rice variety Wuyoudao 4 and E2 aromatic rice variety Songkejing 134 with non-aromatic japonica rice varieties Weinong 101 and Dongfu 114,respectively.The F₂ generation of the hybrid population was screened and identified for aroma genes,and the identification results were consistent with 100%,which verified the accuracy of KASP molecular markers.The 620 japonica rice parents created and collected by our laboratory were further identified by KASP markers,and 248 aromatic rice materials were screened.It was found that these parents were all Badh2-E7 type genes.At the same time,the existing 1 220 F₇ generation materials with E7 aromatic rice as parents were genotyped,including 293 aromatic rice materials,906 non-aromatic rice materials and 21 heterozygous rice materials.Five aromatic and non-aromatic rice materials were randomly selected for sequencing verification.The sequencing results verified that the KASP molecular marker typing was correct.This study provided new markers and materials for breeding new fragrant rice varieties in cold regions by using molecular marker assisted selection.

Waterlogging stress severely impacts soybean production.To investigate the effects of waterlogging stress on soybean growth and development and elucidate its response mechanisms,this study employed four soybean germplasms cultivated in the Huaibei region as experimental materials.Through simulated waterlogging treatment at the V2 stages,combined with yield analysis,physiological indices,and transcriptomic profiling,the study explores the waterlogging tolerance mechanisms in soybean.The results demonstrated that waterlogging stress significantly reduced dry matter accumulation and nitrogen uptake efficiency in the waterlogging-sensitive cultivar Xudou 18,resulting in a yield loss of 30.23%.In contrast,the waterlogging-tolerant cultivar Huaidou 13 exhibited stronger adaptability with only a 16.77% yield loss,attributed to stabilized root nitrogen absorption and maintained root dry matter accumulation.Transcriptomic analysis revealed that differentially expressed genes(DEGs)in Huaidou 13 under waterlogging stress were significantly enriched in pathways that related to the biosynthesis of secondary metabolites,photosynthetic systems,and antioxidant activity.Conversely,DEGs in Xudou 18 were predominantly associated with hormone transduction,photosynthesis,and peroxisome-related pathways.A total of 148 genotype-specific DEGs were identified between cultivars with contrasting waterlogging tolerance,primarily enriched in photosynthesis,secondary metabolite biosynthesis,and fatty acid metabolism.Identified transcription factors included members of the AP2/ERF-ERF,C3H,ARR-B,NAC,and WRKY families.In summary,transcription factors or genes in secondary metabolic pathways that may be related to waterlogging tolerance were screened.

Taking the safflower variety Yuhonghua No.1 as the research object,it adopted the planting method of two harvests of safflower and corn per year was adopted,and six fertilization treatments were set up:blank(T1),straw returning to the field(T2),straw+Bacillus subtilis bacterial fertilizer(T3(125 kg/ha),T4(250 kg/ha)),straw+Bacillus mucilaginosus bacterial fertilizer(T5(125 kg/ha),T6(250 kg/ha)).The influence of straw combined with bacterial fertilizer on the yield of safflower and soil nutrient availability was analyzed.The results showed that the continuous application of straw combined with bacterial fertilizer in 2023 and 2024 had a significant impact on the soil stoichiometric characteristics and yield.In terms of soil stoichiometric characteristics,the T5 treatment was the best for increasing total soil nitrogen(TN),the T3 treatment was the best for increasing total soil phosphorus(TP)and total soil potassium(TK),and the T4 treatment was the best for increasing soil organic matter(SOM).There were significant differences in soil N∶P,K∶P,and N∶K between two years,but the increase and decrease patterns were basically the same.The limiting factors for each treatment were all N and P elements.In terms of output,the T4 treatment was the best for increasing the filament yield,increasing by 62.53% and 52.99% respectively in two years.The T4 and T5 treatments had remarkable increasing effects on the output of seed yield.The output of T4 increased by 7.51% and 8.81% respectively in the two years,while that of T5 increased by 8.28% and 8.59%.There was an extremely significant positive correlation between soil TN,N∶P,N∶K,filament and seed yield,and there was a significant or extremely significant positive correlation between soil TN,SOM and filaments yield.Overall,among the two consecutive years of straw combined with microbial fertilizer application,the T3 treatment had the best effect in improving soil nitrogen,phosphorus,potassium and organic matter.The soils of each treatment were all limited by N and P elements.The T4 treatment had the most significant effect in increasing the comprehensive yield of filament and seed.

Anthocyanin synthase(ANS)is a key enzyme in the anthocyanin biosynthesis pathway in plants,catalyzing the conversion of colorless leucoanthocyanidins into colorful anthocyanins such as red,orange,and blue.To investigate the molecular mechanism of ANS gene in leaf coloration regulation in chicory(Cichorium intybus),bioinformatics analysis of ANS was first conducted.Then,the red heading C.intybus(Indiou)and green forage variety(Puna)were selected as the materials to clone the C.intybus ANS (CiANS)gene.Differences in amino acid sequences and protein structures of CiANS between the two materials were analyzed.Finally,tissue-specific expression of the ANS gene and subcellular localization of CiANS were characterized,and prokaryotic expression of the CiANS was performed.The results showed that CiANS shared the closest phylogenetic relationship with ANS from lettuce(Lactuca sativa)and endive(C.endivia).Motif analysis revealed that the protein motifs of CiANS were relatively conserved across different plant species.Cloning results indicated that the full-length ANS gene in both chicory varieties was 1 068 bp,encoding 355 amino acids with 9 divergent residues,though no significant differences were observed in predicted tertiary structures.qRT-PCR results demonstrated that CiANS was expressed in all tissues,with the highest expression level in leaves,and its expression in red heading Indiou was significantly higher than in green Puna.Subcellular localization revealed that CiANS protein was localized in both the cytoplasm and nucleus.After prokaryotic expression,the induced CiANS protein exhibited a molecular weight of 45 ku,consistent with the predicted size.In conclusion,the observed leaf color variation in C.intybus is likely associated with differential expression levels of the CiANS gene.This study provides theoretical insights for elucidating the molecular regulatory network of Cichorium intybus leaf coloration and genetic improvement of anthocyanin metabolism.

To investigate the function of tachykinin 3 receptor gene(TACR3)and the expression and distribution of its encoded product neurokinin B receptor(NK3R)in the oviduct and uterus of Ganjia Tibetan sheep,hypothalamus,oviduct and uterus tissues during estrous cycle and anestrus of Ganjia Tibetan sheep were taken as samples.The TACR3 gene coding sequence(CDS)was cloned by nested PCR.The biological function of TACR3 interacting proteins was predicted using STRING protein interaction database,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).Real-time Quantitative PCR(qRT-PCR),Western Blot and immunohistochemistry(IHC)were used to analyze the distribution and expression of TACR3 gene and NK3R in the oviduct and uterus during estrous cycle and anestrus of Ganjia Tibetan sheep.The results showed that the total length of the open reading frame of TACR3 gene in Ganjia Tibetan sheep was 1 395 bp,encoding 464 amino acids,and no signal peptide structure was found,with seven transmembrane structures.TACR3 may participate in biological processes such as neuropeptide signaling pathway,GnRH signaling pathway and calcium signaling pathway.The NK3R protein encoded by TACR3 was mainly expressed in the mucosal epithelium,ciliary cells,secretory cells of oviduct,uterine glands and stromal cells of uterus.In oviduct,the expression level of TACR3 mRNA was the highest in the metestrus,and the expression level of NK3R protein was the highest in the estrus.The expression levels of TACR3 mRNA and protein in uterus were the highest in the metestrus.The results showed that TACR3 was conserved in animal evolution,and TACR3 mRNA and protein were expressed in oviduct and uterus tissues at different estrus periods,which may play an important role in the reproductive process of Ganjia Tibetan sheep.

In order to elucidate the molecular mechanism of flour-quality regulation at the transcriptional level,wheat mutant varieties A94 and A261 were obtained by ⁶⁰Co-γ irradiation mutagenesis of Shimai 15,and the developing seeds at days 7,14,21 and 28 after flowering were subjected to transcriptome sequencing,and water absorption,dough development time,stable time,softening degree and farinogram index of the three varieties were measured.The results showed that farinogram parameters of two mutant varieties were better than those of Shimai 15.Twenty-four samples were sequenced,and after the data were filtered and annotated,a total of 26 714 genes were identified.With respect to Shimai 15,there were 1 042,258,301 and 366 differentially expressed genes(DEGs)at four time points in A94,respectively,and there were 2 178,248,117 and 959 DEGs in A261,respectively.KEGG enrichment showed that DEGs were mainly involved in pathways of starch and sucrose metabolism,protein processing in endoplasmic reticulum.GO enrichment showed that DEGs were mainly involved in glycogen biosynthesis processes,amyloplast and other biological processes.By analyzing the differential genes in these pathways and biological processes,it was hypothesized that the reasons for the improved flour quality in the mutant varieties include the up-regulation of genes related to sucrose and starch synthesis and metabolism in the early stages of filling,the degradation of misfolded proteins in the middle and late stages,and the differential expression of genes regulating protein and starch accumulation.

The study aimed to examine the differences in physiological responses to NaCl between these two sweet cherry scion-rootstock combinations,explore the physiological mechanisms governing how different sweet cherry scion-rootstock combinations respond to salt stress,and provide a basis for screening salt-tolerant rootstocks and cultivating sweet cherries in saline-alkali soils.One-year-old plants of the sweet cherry cultivar Meizao(M) grafted onto Gisela 6 (G6) and Colt (KT) rootstocks were used as experimental materials.A pot culture experiment was conducted to simulate NaCl stress,with determinations made on the contents of osmotic adjustment substances,activities of antioxidant enzymes,and photosynthetic gas exchange parameters.The results showed that NaCl treatment promoted the accumulation of osmotic adjustment substances such as soluble sugar,soluble protein,and proline in the leaves of the two sweet cherry scion-rootstock combinations.With the increase in NaCl treatment concentration,the soluble sugar content of M/G6 increased first and then decreased,while that of M/KT continued to increase.Under the 150 mmol/L NaCl treatment,the increase in proline content of M/KT (6.11 times) was significantly higher than that of M/G6 (1.74 times).The contents of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) increased with the increase in NaCl treatment concentration.However,the peroxidase (POD) activity of M/G6 reached its peak at 100 mmol/L,which was significantly higher than that of M/KT.With the increase in NaCl treatment concentration,the net photosynthetic rate (Pn) and stomatal conductance (Gs) of the leaves of the two sweet cherry scion-rootstock combinations gradually decreased,while the intercellular CO₂ concentration (Ci) gradually increased.Non-stomatal limitation was the main factor for the decrease in Pn of the leaves of the two scion-rootstock combinations.M/G6 showed better salt tolerance under moderate salt stress (100 mmol/L),alleviating oxidative damage by relying on the accumulation of soluble sugar and the enhancement of POD activity.M/KT maintained osmotic balance through the accumulation of proline and the increase in SOD activity under high salt stress (150 mmol/L).Based on comprehensive evaluation,in actual production,M/KT can be considered as a reference option for slightly saline soil,and M/G6 is recommended for moderately saline soil.

The aim of this study was to clone the silent mating type information regulator 2-related enzyme 7(SIRT7)gene,detect its expression pattern in different tissues and investigate its effect on milk fat synthesis in mammary epithelial cells of dairy goats.The results showed that by designing the primers of SIRT7 gene,the sequence of SIRT7 gene in dairy goat was successfully cloned,with 1 376 bp and a coding region of 1 203 bp,encoding 400 amino acids and showing the highest homology with sheep.SIRT7 was a hydrophilic protein,free of signal peptides and transmembrane structures,with secondary structures such as irregular coils,α-helices,extended chains and β-turns.The relative expression of SIRT7 gene was the highest in the liver tissues of dairy goats,and it also had certain expression levels in other tissues by qRT-PCR.The pcDNA3.1-SIRT7 overexpression vector was successfully constructed and transfected into mammary epithelial cells of dairy goats.Compared with the control group,the mRNA level of SIRT7 gene in the overexpression group was up-regulated about 33 times.After SIRT7 gene overexpression,the expression of stearoyl-CoA desaturase-1(SCD1)gene was significantly decreased,and the expression levels of cluster of differentiation 36(CD36),diacylglycerol acyltransferase 1(DGAT1),sterol regulatory element binding protein 1(SREBP1)genes extremely significantly reduced.The content of triglyceride in cells was significantly decreased.The results of oil red O staining showed that the accumulation of lipid droplets around the nucleus was significantly reduced.Thus, SIRT7 gene inhibited lipid metabolism in mammary epithelial cells of dairy goats,which laid a foundation for further exploration of the regulatory mechanism of SIRT7 in mammary epithelial cells of dairy goats.

Virus-induced gene silencing(VIGS)is a post-transcriptional gene silencing technique widely used in plant gene function research.However,there are few reports on the establishment of a VIGS system for cabbage in China.The aim of this study was to establish a PCVA/PCVB-mediated gene silencing system using phytoene desaturase(PDS) as an effective visual indicator gene in cabbage.Cabbage,Chinese cabbage,and radish were used as plant materials.The PDS gene was silenced by constructing the PCVA-PDS vector.The PCVA/PCVB-PDS-GFP vector was then generated and transformed into Agrobacterium tumefaciens,which was used to infect cabbage and tobacco epidermal cells via injection and to perform vacuum infiltration in cabbage seedlings.Real-time Quantitative PCR was used to study the applicability of the VIGS system in cabbage,and the system was also applied to another representative Brassicaceae crops-Chinese cabbage and radish.The results showed that green fluorescence could be observed on the cell membranes of cabbage and tobacco leaf cells after infection with Agrobacterium tumefaciens transformed with PCVA/PCVB-PDS-GFP.After 14 d of vacuum infiltration in cabbage seedlings,photobleaching appeared on the new leaves,with the affected area gradually expanding.Real-time Quantitative PCR analysis indicated that the relative expression of PDS homologous genes in the experimental groups decreased 3.2-fold and 1.7-fold compared to the control group,respectively.After infecting Chinese cabbage and radish seedlings,whitening of the leaf veins and some leaves was observed,along with some leaf curling.In conclusion,the photobleaching observed in cabbage leaves after PDS gene silencing demonstrates that the VIGS system effectively replicates and spreads within cabbage plants.The whitening of leaves in Chinese cabbage and radish also indicates that the VIGS system can be applied to other Brassicaceae crops,thus expanding the application scope of this silencing system.The establishment of the cabbage VIGS system provides a theoretical foundation for functional gene studies in Brassicaceae crops.

To explore the expression pattern of potato gene StEXLB1a and provide a theoretical basis for the resistance to potato disease,based on the previous research of the Laboratory of Plant Resources and Molecular Biology at Northeast Agricultural University,the study using the Atlantic variety of potato as material, cloned the StEXLB1a gene, and preliminarily analyzed its bioinformatics, expression pattern, and disease resistance. The results showed that the full-length cDNA sequence of StEXLB1a gene was 768 bp,encoding 255 amino acids.The annotation indicated that it belonged to the extend protein family genes.Subcellular localization showed that the protein was localized to the cell wall.Among different tissues of potato,StEXLB1a gene expression was highest in leaves,followed by roots and stems,and lowest in flowers and tubers.The expression level of StEXLB1a gene changed significantly under various stress treatments,such as hormone(indoleacetic acid,gibberellin,abscisic acid),stress(high temperature,low temperature,salt,drought),fungal disease dry rot(Fusarium avenaceum),bacterial disease soft rot(Erwinia carotovora subsp,Ecc)and bacterial wilt(Ralstonia solanacearum,RS).The overexpressed transgenic potato plants were inoculated with the dry rot pathogen Fusarium avenaceum,and the transgenic plants were more seriously infected than the wild-type plants.The enzyme activities of active oxygen scavenging system in overexpressed plants were significantly changed,POD and SOD activities were inhibited,MDA content was higher than that of wild-type plants,and the damage degree of plants was aggravated.The results showed that transgenic potato plants with overexpression of StEXLB1a had lower resistance to dry rot than those with wild type.

To elucidate the role of the MYL2 gene in muscle development and high-altitude adaptation in yak,and to explore its potential impact on meat quality and production performance,this study conducted cloning and tissue expression analysis of the MYL2 gene in Pamir yak.By cloning its coding sequence(CDS)and performing bioinformatics-based functional prediction and tissue-specific expression profiling,the functional characteristics of the MYL2 gene were further clarified.Additionally,it seeks to analyze and explore the structure and function of the MYL2 gene.Real-time quantitative PCR(RT-qPCR)technology was employed to quantify the expression levels of the MYL2 gene in various tissues of the Pamir yak,including the heart,liver,spleen,lung,kidney,pancreas,and longissimus dorsi muscle.The results indicated that the coding sequence(CDS)length of the Pamir yak MYL2 gene was 501 bp,which encoded a protein of 166 amino acids.Comparative homology analysis revealed that the MYL2 gene of Pamir yak shared the closest genetic relationship with that of the wild yak,exhibiting a similarity of 100%,while the most distant genetic relationship was observed with the chicken.The protein analysis results indicated that the molecular formula of the protein was C₈₄₂H₁₃₁₃N₂₁₉O₂₆₁S₇,with a molecular mass of 18.904 42 ku and a theoretical isoelectric point of 4.831.The instability index was measured at 28.3,suggesting that the protein was stable.Additionally,the protein contained one N-glycan modification potential site and 15 potential phosphorylation sites.Furthermore,its structure did not exhibit a transmembrane domain or signal peptide,and it was primarily localized in the cytoplasmic membrane and cell wall.The secondary structure was predominantly composed of 93 α-helices(56.02%),60 random coils(7.83%),and 13 β-sheets(36.14%).RT-qPCR results showed that MYL2 gene expression was extremely significantly higher in the heart and longissimus dorsi muscle of Pamir yak compared to other tissues,suggesting that this gene may play a key regulatory role in maintaining cardiac function and promoting skeletal muscle growth.Based on protein function prediction,MYL2 is likely involved in the development of cardiac and skeletal muscles by regulating the expression of muscle structural proteins,thereby contributing to the maintenance of locomotor capacity and thermogenic performance under cold,hypoxic plateau conditions.

To determine the relationship between the esters compounds and key enzyme activities and genes that involved in the differences in aromas of oriental melon fruits,the disparities of ester metabolites and the enzyme activities and gene expression characteristics of CXE and AAT were investigated,with the fruits of oriental melon,DX108 and DX3-5,used as experimental materials.The results showed that a total of 644 metabolites were detected in oriental melon fruits,including 114 esters.A total of 108 metabolite variations and 23 different esters were detected in mature fruits.Compared with DX3-5,the types and contents of esters in ripe fruits of DX108 increased significantly,indicating that esters were the key substances in the formation of aroma difference in ripe fruits of oriental melons.K-means clustering analysis showed that isoamyl acetate,ethyl 2-methybuyrate,β-ethyl phenylacetate,p-cresol acetate,hexyl acetate ester,methyl phenylacetate,methyl anthranilate and ethyl caproate were the key esters,which caused the difference of ripe fruit aroma between two oriental melons.In addition,the activities of AAT and CXE in fruits showed opposite trend,suggesting that the coordination of AAT and CXE affected the metabolism of esters.The expression characteristics of AAT and CXE gene family members were different with fruit development,and correlation analysis suggested that CmCXE5,CmCXE6 and CmAAT5 were involved in the metabolism of eight different esters in oriental melon ripe fruits.From those findings,the variations in synthesis and accumulation of eight key esters are important reasons that direct the difference in aroma of two kinds of oriental melon,and CmCXE5,CmCXE6 and CmAAT5 may play a major regulatory role.

A field experiment was conducted from 2023 to 2024 to investigate the response of silage yield in different types of summer maize varieties to various planting patterns.Two varieties,Cangyu 338(CY338,a low-density tolerant variety)and Nongda 372(ND372,a density-tolerant variety),were selected as experimental materials.Four planting patterns were established,including conventional mode(CT),1∶1∶2 mode(1∶1∶2),double plant mode(DP),and 40+80 mode(40+80).Results showed that the dry weight(dry yield)of maize plants and whole ears increased gradually,while the fresh weight(fresh yield)declined during the maize growth period.Significant differences in dry and fresh weight,as well as yield,were observed among the different planting patterns.The dry and fresh weight showed as follows:CT>40+80>1∶1∶2>DP,however,higher plant yield was achieved under DP and 1∶1∶2 patterns.The highest whole ear yield was obtained under CT in 2023 and DP in 2024,followed by the 1∶1∶2 pattern.Additionally,there were significant annual variations in whole ear yield.Dry and fresh weight per plant and whole ear of CY338 were higher than those of ND372,whereas the yield exhibited the opposite trend.A similar changing trend was observed across different years.The response of dry and fresh weight per plant and whole ear to planting patterns indicated that CT resulted in the highest values,while DP yielded the lowest.Except for the dry and fresh weight per plant of ND372 in 2023 under other planting patterns,the 40+80 pattern generally produced higher dry and fresh weights compared to the 1∶1∶2 pattern.In contrast to dry and fresh weight,variety yield exhibited significant variations.The lowest yield for both plant and whole ear was observed under the 40+80 pattern.However,plant yield under the DP and 1∶1∶2 patterns exceeded that of other patterns.Notably,the DP pattern achieved the highest yield,except for ND372 in 2023,which experienced higher precipitation during the experiment.For CY338 in 2024,a year with less precipitation,the DP pattern yielded the highest whole ear production,followed by the 1∶1∶2 pattern,while other varieties showed the highest yield under the CT pattern.In conclusion,planting patterns significantly influence plant and ear characteristics.Corn varieties with high-density tolerance can achieve increased plant and whole ear silage yields.The DP pattern is particularly effective for enhancing plant silage yield(with the 1∶1∶2 pattern being suitable for density-tolerant varieties in rainy years),while the CT pattern is more effective for whole ear silage yield(with the DP pattern being advantageous for low-density-tolerant varieties in the years with less rainfall).

bHLH(Basic helix-loop-helix)gene family is one of the largest plant transcription factor families,binds to cis-acting elements on target gene promoters and plays crucial roles in various physiological processes including growth development,secondary metabolism regulation,signal transduction,and stress response.As one of the world's most important cash crops,cotton is severely affected by stresses such as Verticillium and Fusarium wilt,low temperature,high temperature,salinity,drought,and heavy metal exposure,which significantly impact yield and fiber quality.Meanwhile,with the continuous decline in cotton cultivation benefits,the high-value utilization of by-products such as cottonseeds and cotton secondary metabolites plays a crucial role in improving the comprehensive utilization value of cotton.In this paper,the functions of cotton bHLH transcription factors in cotton fiber development,biotic stress response,abiotic stress response,plant architecture,anther,glandular development,and somatic embryogenesis were summarized.Furthermore,it discusses the application prospects of in-depth analysis of the biological functions of bHLH transcription factors in enhancing the utilization value of cotton by-products,such as cottonseed oil,cottonseed protein,gossypol,anthocyanins,cottonseed vitamins,and nectar,in order to provide theoretical reference for further clarifying the functions of bHLH transcription factors in cotton growth and development,as well as improving the comprehensive utilization value of cotton by-products.

To investigate the molecular mechanisms of the potato miR7997 family in response to Phytophthora infestans infection,this study analyzed the sequence characteristics,target gene prediction,expression patterns,and stress-responsive expression dynamics of Stu-miR7997 and its targets using the potato cultivar Desiree.The results revealed that the potato miR7997 family comprised three members(Stu-miR7997a/b/c)distributed across two chromosomes,with Stu-miR7997a/b sharing identical mature sequences.All precursor sequences formed canonical stem-loop secondary structures,with minimum folding free energies ranging from -37.00 to -49.50 kcal/mol,and mature sequences were located on the 5' arm.Stu-miR7997c exhibited distinct sequence length and functional element distribution compared to Stu-miR7997a/b.Promoter analysis identified light-responsive,hormone-responsive,transcription factor-binding,and stress defense-related cis-regulatory elements;target prediction identified 19 genes predominantly co-regulated by the miR7997 family.Tissue-specific expression profiling showed that Stu-miR7997a/b were highly expressed in stems,while Stu-miR7997c accumulated predominantly in roots.Upon P.infestans infection,all miR7997 members were significantly downregulated,whereas their target genes-including the transcription factor MYB92 gene,NAD(P)H-quinone oxidoreductase gene,and pectin lyase gene-were markedly upregulated.These findings suggest that the Stu-miR7997 family may indirectly modulate potato disease resistance by negatively regulating target genes.This study provides a theoretical foundation for further exploration of the miR7997 family in potato late blight resistance.

The aim of this study was to explore the reproductive adaptability of PEDV in porcine intestinal epithelial cells(IPEC-J2)and to evaluate the inhibitory effect of Polygonum cuspidatum water extract on it in vitro.The effects of different trypsin concentrations and different time on the activity of IPEC-J2 cells and the expression of PEDV N protein in cells were evaluated by cytotoxicity assay and Western Blot assay.The virus titer,PEDV N gene transcription level and N protein expression were detected by TCID₅₀,Real-time Fluorescence Quantitative PCR and Western Blot.The effect of water extract of P.cuspidatum on the proliferation activity of IPEC-J2 cells was detected by CCK-8 method.Further,the inhibitory effect of P.cuspidatum water extract on the expression of PEDV N gene and N protein was detected by Real-time Fluorescence Quantitative PCR and Western Blot experiments,and the effect of P.cuspidatum water extract on each stage of PEDV life cycle was explored.The results showed that when the concentration of trypsin was 2 μg/mL,trypsin had no effect on the cells.At this time,PEDV showed the best reproductive adaptability and reached the highest virulence value after 36 h.When the concentration of P.cuspidatum water extract was lower than 100 μg/mL,there was no cytotoxicity at 12,24 h.The water extract of P.cuspidatum significantly inhibited the expression of N gene under Co-treatment and Post-treatment,and significantly inhibited the expression of PEDV N gene mRNA and N protein in a dose-dependent manner.The water extract of P.cuspidatum had a significant inhibitory effect on PEDV N gene mainly in the replication and proliferation stage of the virus life cycle,and had no significant inhibitory effect in other stages.In summary,2 μg/mL trypsin had the best adaptability to PEDV reproduction,and the water extract of P.cuspidatum had a significant inhibitory effect on PEDV in vitro,which mainly acted on the replication and value-added stage of PEDV life cycle.

Fat storage inducing transmembrane protein 2 (FITM2) plays an important role in regulating lipid storage and skeletal muscle energy metabolism.To scan the polymorphisms of the FITM2 gene in sheep and investigate its relationship with growth traits in Hu sheep,the experiment selected 1 128 healthy and disease-free,genetically well-documented Hu sheep as the experimental group,and used the FITM2 gene in sheep as the research object.First,the expression differences of the FITM2 gene in different tissues were analyzed using qPCR technology.Then,the genetic polymorphisms of the FITM2 gene in sheep were detected using PCR amplification,Sanger sequencing,and AQP (allele-specific quantitative PCR-based genotyping assay).Finally,the association between the genetic polymorphisms and growth traits was analyzed.The research results showed that the FITM2 gene was expressed in different tissues of Hu sheep,with the highest expression level in the tail fat and significantly higher than that in other tissues.The gene polymorphism detection results showed that there was a C>T mutation at position 72704027 in the first intron of the FITM2 gene.The results of correlation analysis showed that the 100,120 d body weight,140,160 d body height,80,120,140,160 d body length,and 100 d chest circumference of CC genotype individuals were significantly higher than those of TT genotype individuals.In summary,the FITM2 gene g.72704027 C>T mutation site in sheep can be used as a candidate molecular marker related to growth traits in Hu sheep,providing a theoretical basis for the genetic breeding work of Hu sheep.

It is of great significance to study the effects of brackish water sprinkler irrigation on the growth and yield of summer maize to alleviate the shortage of fresh water resources,promote the utilization of unconventional water resources and guarantee the production of summer maize.Field trial was conducted for two consecutive years in the central plain area of the Heilonggang Basin,Hebei Province.It aimed to investigate the effects of three sprinkler irrigation treatments(fresh water,2 g/L brackish water,and 3 g/L brackish water)on root distribution,photosynthetic performance,and grain yield in summer maize.Results showed that the average dry root weight density of 0-60 cm soil layer in the whole growth period of 2 g/L brackish water sprinkler irrigation was reduced by 7%-9% than fresh water irrigation but with no significant difference,and the average root dry weight density of 3 g/L brackish water sprinkler irrigation was significantly reduced by 19%-26%.Compared with fresh water irrigation,the 2,3 g/L brackish water sprinkler irrigation both had adverse effects on the photosynthetic characteristics of leaves,especially the 3 g/L brackish water sprinkler irrigation,the net photosynthetic rate,stomatal conductance and light use efficiency were significantly reduced by 29%-37%,40%-49% and 29%-35%,respectively,which seriously affected the photosynthetic capacity of the leaves.In 2020,the leaf area index of 2 g/L brackish water sprinkler irrigation was reduced by 15% compared with that of fresh water sprinkler irrigation but with no significant difference,and the leaf area index of 3 g/L brackish water sprinkler irrigation was significantly reduced by 28%.In 2021,there was no significant difference in leaf area index between different treatments during the whole growing period.There was no significant difference in plant height between different treatments during the whole growth period of two years.2 g/L brackish water sprinkler irrigation had no significant effect on yield of summer maize in both years,and the yield of 3 g/L brackish water sprinkler irrigation was significantly reduced by 9% in 2020.No significant differences were observed in ear count,kernel number per ear,or 100-grain weight across the irrigation treatments.In summary,the salinity of brackish water sprinkler irrigation less than 2 g/L can guarantee the growth and yield of summer maize in this area.

This study was carried out to understand the effect of different light quality treatment on the growth,photosynthesis and antioxidant enzyme activities of tomato seedlings in artificial light plant factory.The trial was conducted at a light intensity of 200 μmol/(m²·s),with blue LED light(B)as control.Green light(G),red light(R),far red light(Fr),or ultraviolet light(UV)with consistent light intensity was added on the basis of blue light to clarify the growth,photosynthetic and antioxidant enzyme activities of tomato seedlings under different light quality combinations.The results showed that compared with control,the height of tomato seedlings under B+Fr treatment was significantly improved,increased by 34.54%.B+R treatment significantly increased the stem diameter and dry and fresh weight of tomato seedlings by 39.27%,25.58% and 22.20%,respectively compared with B.The intercellular CO₂ concentration and stomatal conductance(Gs)of tomato seedlings under B+G treatment were higher than other treatments.Besides,B+R treated seedlings had the highest effective photochemical quantum yield of PS Ⅱ,the actual photochemical quantum yield of PSⅡ and the photochemical quenching coefficient,increased by 4.30%,0.97% and 1.87%,respectively compared with the control.Furthermore,B+G,B+R treatment significantly increased the antioxidant enzymes activity,and significantly decreased the content of malondialdehyde(MDA),reactive oxygen species and hydrogen peroxide in tomato seedlings.Thereinto,MDA content of B+R treated seedlings was 50.64% lower than B treatment,while SOD,POD and APX activities were 15.44%,25.00% and 21.65% higher than B treatment,respectively.Taken together,B+R treatment was not only conducive to the cultivation of strong tomato seedlings,but also helped improving PSⅡ activity of leaves,thus enhancing photosynthetic performance.Meanwhile,B+R light treatment could also maintain a high level of antioxidant enzyme activity in tomato plants and improved the antioxidant capacity of tomato plants.

Microtubules and microtubule-binding proteins in cotton fibroblasts play an important role in regulating intracellular material transport and morphogenesis.The TOG domain is an important functional component of several protein families that regulate microtubule dynamics,and CLASPs are a unique class of microtubule-binding proteins in plants,whose expression is very important for the dynamic adjustment and functional exercise of microtubules.In order to study the relationship between Togaram1 gene and cotton fiber development and fiber quality formation,GhTogaram1 and GbTogaram1 genes were cloned from upland cotton Line 9 and sea island cotton Xinhai 16,and their bioinformatics analysis was conducted.Meanwhile,qRT-PCR was used to analyze the expression patterns of Togaram1 gene in different materials and different fiber development stages.The results showed that the total length of CDS sequence of GhTogaram1 and GbTogaram1 was 918 bp,which encoded 305 amino acids,but there were nucleotide and amino acid sequence differences between upland cotton and sea island cotton.Bioinformatics analysis showed that Togaram1 protein was a hydrophilic protein with TOG domain and CLASP-N-terminal domain,and was localized in the nucleus.The phylogenetic tree and multiple sequence comparison showed that GhTogaram1 and Gossypium barbadense belonged to the same cladisticbranch,the similarity was 99.34%,and the closest relation was Gossypium mustelinum.The results of qRT-PCR showed that the relative expression levels of Togaram1 gene were significantly different at different stages of fiber development(0,5,25 d)between the two materials.In addition,there were significant differences in the fiber development at 5 and 10 d of Togaram1 gene in the extreme offspring of the HL population,and the expression level of HL-78 was the highest in the short fiber line.In conclusion,Togaram1 gene was predominantly expressed in the fibers of different cotton materials 5 d after flowering,indicating that Togaram1 gene may play a role in cotton fiber elongation.With the increase of flowering days,the expression of Togaram1 gene decreased,which may be due to the influence of brassinsterol.The preliminary functional verification of Togaram1 gene in cotton fiber laid a foundation for the subsequent study of Togaram1 gene in cotton fiber development.

Hedgehog(Hh)signaling pathway plays an important role in the growth and development of various organisms,and Smoothened(Smo)protein,as a key receptor of Hh signaling pathway,is a core regulator of Hh signal transduction.In order to investigate the function of Smo protein and its regulatory mechanism in honey bees,and to deeply analyze the molecular mechanism of olfactory signal transduction in honey bees,the Smo protein was expressed and purified by using the Escherichia coli BL21 expression system,and the purified Smo protein was used as the antigen to immunize New Zealand white rabbits,and a polyclonal antibody against Smo protein with high activity and specificity was prepared.The expression localization of Smo protein in the antennae of Apis mellifera was further detected by immunofluorescence technique.The results showed that the recombinant Smo protein was enriched at 83.72 ku after induced expression in E.coli,which was consistent with the predicted molecular weight size,indicating that the induced expression was successful;the results of SDS-PAGE analysis showed that,after enrichment and purification,a high concentration of recombinant Smo protein eluate was obtained,indicating that the Smo protein was purified well.The results of ELISA assay showed that the antibody potency of the prepared Smo protein reached 1∶364 500,showing high immunoreactivity.Immunofluorescence showed that Smo protein was widely expressed in the antennae of western honey bees,especially in the hairy sensilla.The polyclonal antibody to Smo protein was successfully prepared with high purity,high potency and specificity,and the localization of Smo protein in the antennae of western honey bees was clarified.

To study the effect of silicon fertilizer on the coupled process of methane (CH₄) anaerobic oxidation and nitrous oxide (N₂O) reduction in warming paddy fields,it could provide scientific evidence for exploring new ways to reduce greenhouse gas emissions in paddy fields.Four field treatments were set up,including no silicon application at ambient nighttime temperature (CK),no silicon application at nighttime warming (NW),silicon application at ambient nighttime temperature (Si),and silicon application at nighttime warming (NW+Si).Surface soils were collected of above-mentioned treatments,and were conducted anaerobic cultivation under different gas (¹³CH₄,¹³CH₄+N₂O,N₂O) addition conditions.Using isotope labeling method combined with quantitative PCR technology,it aimed to study the effect of warming and silicon application on the CH₄ anaerobic oxidation driven by N₂O reduction in paddy fields.The CH₄ oxidation rate driven by N₂O reduction in NW paddy fields was 3.99 nmol/(g·d),which was not significantly different from that in CK paddy fields;the CH₄ oxidation rate in Si paddy fields was only 1.90 nmol/(g·d),significantly lower than that in CK and NW+Si paddy fields.This indicated that silicon application had an inhibitory effect on the coupling process of CH₄ anaerobic oxidation and N₂O reduction in paddy fields at ambient temperature,while nighttime warming had no significant effect on this process.Under the addition of ¹³CH₄+N₂O treatment,the number of mcrA genes in four paddy fields reached 3.53×10⁷-8.19×10⁷ copies/g,which was 24%-35% higher than that in the ¹³CH₄ treatment.However,there was no difference in the anaerobic pmoA gene between these two treatments,indicating that the anaerobic oxidizing archaea M.nitroreducens might directly participate in the coupling process of ¹³CH₄ oxidation and N₂O reduction.The copy number of nosZ Ⅱ gene was significantly positively correlated with CH₄ oxidation rate driven by N₂O reduction,indicating that nosZ Ⅱ microorganisms might play an important role in this CH₄ oxidation process.Unlike paddy fields at ambient temperature,silicon application had no inhibitory effect on the coupled process of CH₄ anaerobic oxidation and N₂O reduction in paddy fields with nighttime warming.The CH₄ anaerobic oxidating archaea M.nitroreducens and nosZ Ⅱtype N₂O reducing bacteria may be involved in this coupling process.

In order to explore the effects of different canopy light environments on apple mesophyll conductance and its key parameters and photosynthetic biochemical parameters,the open-center Fuji apple trees were used as experimental materials.The canopy light environment,photosynthetic biochemical parameters and leaf tissue structure of apple trees were measured after thinning,and the apple trees without thinning were used as controls(CK).Results showed that,the peak and mean value of daily maximum and averaged photosynthetically active radiation(PAR)increased by 23%,166%,and 43%,123%,respectively.Compared to the CK,product of light absorption coefficient and light energy partitioning ratio(α·β)and dark respiration rate under light(R_d)of treatment increased by 84.62%,56.00% respectively,while apparent CO₂ compensation point(C_i^*)value decreased by 6.67%.The average value of mesophyll conductance(g_m)calculated by quantitative α·β value was 9.89% higher than that of the CK,and the maximum and average values of mesophyll conductance(g_m')calculated by empirical α·β value(0.425)were 20.65%,39.38% higher than those of the CK,respectively.The g_m ' of the treatment and the CK was higher than that of the g_m,and the g_m' of the treatment and the CK was 54.33%,21.67% higher than the average value of the g_m,respectively.V_cmax derived from P_n-C_i(neglecting mesophyll conductance),P_n-C_c'(using empirical α·β)and P_n-C_c(using quantified α·β)response curves demonstrated significant differences(V_cmax-Ci and V_cmax-Cc' underestimated by 40.59%,19.85% in comparison with V_cmax-Cc in treatment,respectively),and treatment was significantly higher than CK(V_cmax-Ci,V_cmax-Cc' and V_cmax-Cc increased by 11.21%,5.85%,10.69%,respectively);J_max and V_tpu were similar to V_cmax.The anatomical structures of leaves showed that the mean values of palisade tissue(PT)thickness and spongy tissue(ST)thickness of thinning treatment significantly increased respectively by 43.95%,43.31%,in comparison with those of the CK.The area and perimeter of PT and ST in thinning treatment were respectively 43.60% and 310.86% with 34.73% and 115.82% higher than those in CK.In summary,after thinning treatment,the light condition of apple canopy was improved,the leaf structure was changed,and the photosynthetic efficiency of the plant was improved.

To investigate the effects of combined nitrogen and selenium fertilizer application on non-structural carbohydrates, yield, and quality of oil flax, this study utilized Longya 11 as the test material.A two-factor split-plot field experiment was conducted to examine the effects of nitrogen fertilizer(N0:0 kg/ha,N1:75 kg/ha,N2:150 kg/ha)and selenium fertilizer(Se:0 g/ha,Se1:30 g/ha,Se2:60 g/ha)on oil flax non-structural carbohydrates,grain yield and quality of oil flax under dryland conditions.The results showed that the content of soluble sugar of stems and leaves decreased gradually with the advancement of growth,while the content of starch of stems was the opposite.Under the same selenium fertilizer level,nitrogen fertilization significantly increased the soluble sugar and starch content of stems and leaves,dry matter accumulation,branch number,valid fruits,seed number grain,yield,and seed protein and oleic acid content,and the N2 level had the highest value,and the increase was greater at maturity stage.Under different nitrogen application levels,the combined application of N1 or N2 and selenium fertilizer significantly increased the soluble sugar and starch content of stems and leaves,the dry matter accumulation,branch number,valid fruits,seed number,1000-seed weight,grain yield,protein,oil content,lignans,and oleic acid content.Among the treatments,N2Se1 treatment significantly increased soluble sugar,starch content of stems and leaves and dry matter accumulation at maturity,and the valid fruits,seed number,and the 1000-grain weight were significantly higher than those of other treatments,which were 12.35%,3.89%,4.18% significantly higher than those of N2Se0 treatment,respectively.The oleic acid content of seed of N2Se1 was significantly higher than that of other treatments by 2.44%—25.64%,and the seed yield reached 1 700.50 kg/ha.The results showed that the combined application of 150 kg/ha nitrogen fertilizer and 30 g/ha selenium fertilizer could significantly increase the non-structural carbohydrate content and dry matter accumulation of oil flax stems and leaves in the test area,increase seed yield and oleic acid content,and improve quality.

Muscle formation depends on the interplay of various cellular and extracellular signals and factors.This study investigates the regulatory relationship between Myocyte enhancer factor 2A(MEF2A)expression levels and promoter methylation at both tissue and cellular levels,to provide theoretical references for the genetic development of Guanling cattle.Guanling cattle were used as experimental subjects.Initially,the expression levels of the MEF2A in Guanling cattle tissues were analyzed in conjunction with their promoter methylation status.Subsequently,overexpression and interference vectors of the MEF2A gene were successfully transfected into primary myoblasts of Guanling cattle,and the effects of MEF2A expression level changes on promoter methylation levels were analyzed.The results showed that the MEF2A expression levels in various tissues of young Guanling cattle were higher than those in adult cattle,while the methylation rate of MEF2A in young cattle tissues was lower than that in adult cattle,with DNMT1 expression trends consistent with this.Additionally,increased MEF2A expression led to a decrease in its methylation rate,whereas decreased MEF2A expression resulted in an increase in its methylation rate.This study confirms from both tissue and cellular perspectives that the expression level of the MEF2A gene in Guanling cattle is negatively correlated with its methylation rate,providing a theoretical basis for genetic marker-assisted genetic improvement of cattle.

To identify the suitable sowing date and planting density for the synergistic improvement of yield and quality of strong gluten wheat,the strong gluten wheat variety Kexing 3302 was used as the experimental material.Two sowing dates(October 18(S1)and October 28(S2))and five densities(basic seedlings of 180×10⁴ plants/ha(D180),225×10⁴ plants/ha(D225),270×10⁴ plants/ha(D270),315×10⁴ plants/ha(D315),and 375×10⁴ plants/ha(D375))were set.Field experiments were conducted in 2021—2023 to investigate the effects of sowing date and density on physiological indicators,yield,and quality of wheat plants.The results showed that the relative chlorophyll content(SPAD)of wheat leaves generally decreased with the increase of planting density,and reached its maximum at most growth stages under low-density treatment.With the delay of the sowing date,the relative chlorophyll content of leaves showed an upward trend,with S1 sowing date being lower than S2.As planting density increased,the normalized difference vegetation index(NDVI)generally showed an upward trend,and the NDVI value at S2 sowing date was significantly lower than that at S1 sowing date.The number of spikes and grain yield decreased with the delay of the planting density,but increased with the increase of planting density,with the highest yield under D375 treatment.With the delay of the sowing date,both protein content and wet gluten content generally showed a slight downward trend,but the grain protein content of all treatments was above 13.5%.The total starch content of wheat grains generally decreased with the increase of density at S1 sowing date,but increased with the increase of density at S2 sowing date.In addition,the peak viscosity,trough viscosity,final viscosity,and breakdown value generally increased with the increase of density under the two sowing dates.Parameters such as dough stability time and water absorption rate did not exhibit obvious regular pattern under different sowing dates and planting density,but all met the standards for strong gluten wheat.In summary,at sowing date S1,the planting density of Kexing 3302 at 315×10⁴—375×10⁴ plants/ha can achieve high quality and high yield.

Drought during the grain filling period can seriously affect the later growth and development of foxtail millet plants,as well as the morphogenesis,yield,and quality of grains.To analyze the effects of drought during the grain filling period on the later growth and development of foxtail millet,as well as the morphogenesis of grains,and to provide a theoretical research basis for the molecular mechanism of foxtail millet starch regulation in response to drought stress,it investigated the effects of different degrees of drought during the grain filling period on the agronomic and grain quality traits of foxtail millet using Jingu 21 as the material.Simultaneously,Real-time Quantitative PCR was used to analyze the response of key genes SiAGPase1(Glucose pyrophosphorylase)and SiSSS1(Starch synthase)to drought during starch synthesis.The results showed that drought had significant effects on fresh weight per plant,fresh weight per root,fresh weight per panicle,panicle weight,grain weight per spike,grain weight per thousand grains,main stem diameter,subpanicle internode length,chlorophyll content,seed setting rate,spike code density and other agronomic characters.The more severe the stress,the greater the degree of reduction.The analysis of grain composition showed that drought can reduce the content of fat,crude protein,peak viscosity and final viscosity.The more severe the stress,the greater the reduction degree.Gene expression analysis showed that the expression levels of SiAGPase1 and SiSSS1 changed sharply during the grain filling period under drought stress.The results revealed that SiAGPase1 and SiSSS1 played an active role in regulating the balance between starch decomposition and starch accumulation in grains under drought stress in order to adapt to plant response to drought stress.

The members of the Aux/IAA gene family encode proteins that regulate various developmental processes in plants,such as embryogenesis and seed development,by modulating the auxin (IAA) signal transduction pathway.To explore the role of Aux/IAA family genes in jujube embryo development,this study performed bioinformatics analysis and functional verification on the key genes of the Aux/IAA family that affect jujube embryo development,using transcriptome data of Huizao (a jujube variety with high embryo fertility) and Kongfusucui (a jujube variety with low embryo fertility).Results showed that an Aux/IAA family gene sequence with a length of 1 731 bp was obtained via gene cloning,which encoded 362 amino acids.Protein structure prediction indicated that its secondary structure was mainly composed of random coil and that it had a typical conserved domain of the IAA9 protein;thus,it was named ZjIAA9.Homology analysis revealed that ZjIAA9 was closely related to RGQ29_009000 in Quercus rubra and CFP56_014209 in Quercus suber.Transgenic Micro-Tom tomato plants were obtained via Agrobacterium-mediated leaf disc transformation.Determination of auxin content in transgenic Micro-Tom tomato plants showed that the IAA content in the pulp of transgenic plants was higher than that in non-transgenic plants,indicating that ZjIAA9 may regulate auxin biosynthesis.Compared with wild-type plants,transgenic plants showed a seedless or few-seeded phenotype,suggesting that jujube ZjIAA9 may be involved in regulating jujube embryo development.

To clarify the effect of short-day photoperiod induction on the metabolic material changes in adzuki bean leaves,the late maturing variety Jihong 16 was used,and 10 h light /14 h dark short-day photoperiod induction was set.The group(10h14d)of short-day photoperiod induction and control group(CK)were studied using ultra-high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).The results demonstrated that a total of 128 significantly differential metabolites were obtained by quantitative testing(103 substances were up-regulated and 25 substances were down-regulated),including 11 metabolites,such as flavonoids,amino acids and derivatives,organic acids,phenolic acids etc,the number(proportion)were 49(38.28%),26(20.31%),12(9.38%),10(7.81%),all above 10%.KEGG enrichment analysis found that 128 metabolic substances were enriched in 49 metabolic pathways.Moreover,significant differential top 20 metabolic pathways were mainly enriched in glucosinolate biosynthesis,aminoacyl-tRNA biosynthesis,2-oxocarboxylic acid metabolism,isoflavonoid biosynthesis,cyanoamino acid metabolism,biosynthesis of amino acids,flavonoid biosynthesis etc.Among the top 10 metabolic pathways with a proportion greater than 10%,the enriched metabolites were mainly acids and ketones.Moreover,most of these metabolites were upregulated.In summary,ketone and acid-related metabolites are considered to be main metabolites in response to short-day photoperiod induction.This provides a theoretical basis for optimizing the planting structure and improving the yield and quality of adzuki bean.

The objective of this study was to identify differentially expressed mRNAs,miRNAs,and lncRNAs between different tail types of sheep in their tail fat tissues,and to construct a critical ceRNA(competing endogenous RNAs)regulatory network involved in the regulation of fat deposition in the tails of sheep.Three 2-year-old(24-month-old)Dorper sheep and three Tianjin large-tail sheep were randomly selected.Tail fat tissues were collected for H&E staining and whole transcriptome sequencing.Bioinformatics approaches were employed to screen for differentially expressed RNAs,predict target genes,and conduct functional enrichment analyses.The regulatory network and targeting relationships were further analyzed from the perspective of lipid deposition regulation.The results demonstrated that the tail fat cell area of Tianjin large-tail sheep was significantly larger than that of Dorper sheep.There were a total of 1 156 differentially expressed mRNAs between the tail fat tissues of Dorper sheep and Tianjin large-tail sheep(with 403 upregulated and 753 downregulated),72 differentially expressed miRNAs(46 upregulated and 26 downregulated),and 392 differentially expressed lncRNAs(142 upregulated and 250 downregulated).Additionally,216 target genes were predicted from the differentially expressed lncRNAs.These target genes were involved in biological processes related to lipid metabolism,such as the tetrahydrofolate metabolic process,GAIT complex,and coenzyme catabolic process,as well as signaling pathways like glycerolipid metabolism,biotin metabolism,fat digestion and absorption,and the phosphatidylinositol signaling system.The study successfully constructed a ceRNA network closely associated with sheep tail fat deposition by predicting the targeting relationships between differentially expressed mRNAs and lncRNAs and miRNAs.Among them,24 lncRNAs and 6 miRNAs were predicted to regulate genes closely related to lipid metabolism,including PROX1,LPL,and LRP1B.In summary,PROX1,LPL,and LRP1B play a key role in the deposition of tail fat in sheep.This study predicts that four lncRNAs,including MSTRG.9916.7,positively regulate fat metabolism through PROX1,while MSTRG.7563.1 and MSTRG.4729.1 positively regulate fat metabolism through LPL.

Catalase(CAT)plays a critical role in plant resistance against biotic stresses.To investigate the functions of StCAT1 and StCAT3 in potato defense against Alternaria solani infection,this study cloned the cDNA sequences of StCAT1 and StCAT3 via RT-PCR.Sequencing results demonstrated that both genes were 1 479 bp in length,encoding 493 amino acids.Phylogenetic tree construction revealed that StCAT1 shared high homology with SlCAT1 from tomato,while StCAT3 exhibited high homology with both SlCAT3(tomato)and SpCAT3 from wild tomato.Following A.solani infection for 6 days,the expression of StCAT1 and StCAT3 was significantly upregulated by approximately 3.9-fold and 8.7-fold,respectively.A co-silencing vector targeting both StCAT1 and StCAT3 was constructed,and transient co-silencing was achieved using virus-induced gene silencing(VIGS).qRT-PCR screening identified an effective co-silenced potato line(VIGS-3)with suppressed StCAT1 and StCAT3 expression.Upon A.solani inoculation,the silenced line exhibited 42% larger lesion areas compared to the control,indicating that co-silencing StCAT1/StCAT3 compromised potato resistance to early blight.DAB(3,3'-diaminobenzidine)staining further demonstrated that H₂O₂ accumulation in co-silenced leaves was significantly elevated,with staining intensity being 3.8-fold higher than that in the control.These results demonstrate that StCAT1 and StCAT3 regulate H₂O₂ homeostasis to mediate potato resistance against A.solani,providing a theoretical foundation for elucidating the molecular mechanisms of CAT genes in potato-pathogen interactions.

In order to explore the molecular mechanism of the interaction between Bursaphelenchus doui and host plants,an FMRFamide-like neuropeptide gene,named Bd-FLP-12,was cloned from B.doui by using RACE-PCR technique in a specific cDNA library of the anterior end of B.doui.Further,the nucleotide sequence and amino acid sequence of Bd-FLP-12 were analyzed using bioinformatics methods,the gene copy number was identified by Southern Blot,and the developmental expression pattern was investigated by semi-quantitative RT-PCR.The Bd-FLP-12 gene encoded a protein of 90 amino acids with a mature peptide sequence of FLP-12 and a signal peptide sequence but no transmembrane structure,indicating that its encoded protein was a secretory protein.Southern Blot indicated that the Bd-FLP-12 gene was a single copy gene in the B.doui genome,and the RT-PCR result showed that the transcription level of Bd-FLP-12 was lower in the egg stage than in the other stages.In summary,the full-length sequence of the Bd-FLP-12 gene was cloned for the first time in B.doui,and the structure,nature and expression of the gene were characterized,which could serve as a foundation for further study of the gene function.

To investigate the function of StIMPα in potato, this study used the potato cultivar Kexing No.1 as material and successfully cloned the StIMPα2 gene via PCR. Bioinformatic analysis revealed that the coding sequence (CDS) of StIMPα2 had a length of 1 590 bp, encoding a protein containing the typical domains of the IMPα family. Phylogenetic tree analysis indicated that StIMPα2 was most closely related to AtIMPα-1 and AtIMPα-2 from Arabidopsis thaliana, suggesting functional conservation. Furthermore, analysis of the StIMPα2 promoter region identified multiple cis-acting elements associated with responses to biotic and abiotic stresses. To determine its subcellular localization, a StIMPα2-GFP fusion expression vector was constructed and transiently expressed in leaves of Nicotiana benthamiana via Agrobacterium-mediated transformation. Confocal laser scanning microscopy showed that the GFP fluorescence signal was specifically enriched in the nucleus, confirming that StIMPα2 is a nuclear-localized protein. Expression pattern analysis demonstrated that StIMPα2 expression was significantly induced by abiotic stresses such as low temperature, high salinity, and drought, as well as by BTH (benzothiadiazole). For functional validation, StIMPα2 was overexpressed in N. benthamiana via Agrobacterium infiltration, followed by inoculation with Phytophthora infestans. Pathological phenotype analysis showed that compared with the control, the lesion area on leaves overexpressing StIMPα2 was significantly reduced. Meanwhile, Quantitative Real-time PCR detection of P. infestans biomass confirmed a significant decrease in pathogen biomass in StIMPα2-overexpressing plants. In conclusion, these results indicate that StIMPα2 is a nuclear-localized protein induced by various biotic and abiotic stresses, and it enhances resistance to P. infestans by positively regulating plant immune responses.

In order to identify the molecular biological characteristics and expression patterns of Galectin-3 gene in Chinese giant salamander before and after infection,the Galectin-3 gene was amplified by RT-PCR,and its physicochemical properties and protein structure were analyzed by bioinformatics.Real-time Quantitative PCR(qRT-PCR)was used to detect the expression of Galectin-3 in different tissues of Chinese giant salamanders before and after Aeromonas hydrophila.The results showed that the length of Galectin-3 gene CDS region was 351 bp,encoding 116 amino acids,the theoretical isoelectric point(pI)was 5.87,the relative molecular weight was 13.53 ku,and it was a stable hydrophilic protein located in the cytoplasm.Structural analysis showed that Galectin-3 protein contained a GLECT domain.The phylogenetic tree showed that the Galectin-3 amino acid sequence was closely related to Rhinatrema bivittatum.qRT-PCR results showed that Galectin-3 was expressed in all tested tissues of the salamander,and the relative expression level was the highest in the skin.The relative expression levels of Galectin-3 in the liver,muscle,intestines and kidneys of Chinese giant salamander increased significantly and reached the highest value at 12 h of infection with A.hydrophila.The relative expression levels of Galectin-3 in the spleen and skin of Chinese giant salamander increased significantly and reached the highest value at 48 h of infection with A.hydrophila.In conclusion,Galectin-3 gene was actively expressed in the early stage of salamander infected by foreign bacteria,suggesting that Galectin-3 gene may be involved in the immune regulation of salamander.

In order to explore the diurnal variation characteristics and influencing factors of surface CO₂ and N₂O emission fluxes in tobacco fields in central Henan under the management mode of continuous green manure incorporation,and to determine the optimal collection time of the two gases,in 2021,based on a long-term experiment in Maozhuang Science and Education Park of Henan Agricultural University,a 24-hour continuous dynamic observation was carried out by static dark box-gas chromatography every 30 days after tobacco seedlings were transplanted.The daily emission fluxes of CO₂ and N₂O in tobacco fields under nitrogen phosphorus potassium treatment(NPK)and NPK+planting and turning over ryegrass treatment(NPKG)were measured.The results showed that the tobacco field soil in central Henan was the source of CO₂ and N₂O emissions,and the emission trend was similar to the trajectory of atmospheric temperature change,which was high in day and low in night,and the average emission flux of day and night reached a significant difference level.The CO₂ emission flux showed an inconspicuous bimodal pattern at 30 and 60 days after transplanting,and a single peak state of high day and low night at 90 days after transplanting.The N₂O flux showed a single peak state of high day and low night.The CO₂ and N₂O emission fluxes of NPKG treatment were significantly higher than those of NPK treatment.The CO₂ emission fluxes of three typical days were 90 d>60 d>30 d,and the N₂O emission fluxes were 60 d>90 d>30 d.During the observation days,water-filled porosity of soil(WFPS) and 10 cm soil temperature jointly affected the emission rates of CO₂ and N₂O,and the 10 cm soil temperature was significantly or extremely significantly positively correlated with the daily emission fluxes of CO₂ and N₂O.The correction coefficients of daily CO₂ and N₂O emission fluxes at 9:00 and 21:00 were the closest to 1,and there was no significant difference compared with the daily average emission flux.There was a significant positive correlation between CO₂ flux and N₂O flux.Within a certain range,the N₂O flux increased linearly with the increase of CO₂ flux.In summary,continuous green manure returning increased the surface CO₂ and N₂O emission flux of tobacco field,and the best collection time of CO₂ and N₂O was around 9:00 and 21:00.During the three sampling days of tobacco growth period,the surface CO₂ emission flux of NPK treatment and NPKG treatment was the highest at 90 days after transplanting,followed by 60 days.The N₂O emission flux was the highest at 60 days after transplanting,followed by 90 days.