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.

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 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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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 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.

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.

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.

The hexameric Paf1 (RNA polymerase Ⅱ associated factor 1) complex is a crucial transcription regulator in eukaryotes.Paf1-regulated expression of specific genes in plants is closely related to diverse biological processes including growth,development,and stress responses.In order to get information on the responses of Paf1 to abiotic stresses in common wheat,homologous sequence searches were performed to identify all of the genes encoding each of the Paf1 subunits in the wheat genome.mCherry fusions of the wheat Paf1 subunit proteins were expressed in protoplasts and tobacco leaves for determination of protein subcellular localization by fluorescence microscopy.qRT-PCR assays were conducted to profile the expression of wheat Paf1 subunit genes in response to different abiotic stresses.The results showed that,in wheat,five of the Paf1 subunits,TaVIP3,TaVIP4,TaVIP5,TaVIP6,and TaPHP,were each encoded by one set of homeologous genes while the sixth subunit TaVIP2 was encoded by two sets.Plant VIP2 sequences had an N-terminal proline-rich region with variable length,and wheat TaVIP2 sequences had an additional glutamine-rich region.Protein subcellular localization assays revealed the nuclear localization of TaVIP2,TaVIP4,TaVIP5,and TaVIP6 proteins and the nuclear and cytoplasmic localization of TaVIP3 and TaPHP proteins.Gene expression analyses revealed similar tissue-dependent constitutive expression variations and similar stress-induced expression patterns of wheat Paf1 subunit genes.These genes coordinately responded to the stress of high temperature by expression upregulation and to the stresses of salt and drought by expression downregulation.Collectively,our results suggested the involvement of expression regulation of Paf1 subunit genes in the responses of wheat to abiotic stresses.

In order to investigate the effects of conservation tillage techniques on soil nutrient and enzyme activity indicators,as well as ecological stoichiometry characteristics in black soil areas,this study used a 3-year positioning experiment method,with tillage (A-A) as the control,to investigate the changes in total nutrients soil organic carbon(SOC),total nitrogen (TN),total phosphorus(TP) and enzyme activities of β-D-glucosidase(βG),leucine aminopeptidase(LAP),N-1,4-acetylglucosidase(NAG),acid phosphatase(ACP)and their ecological stoichiometry characteristics in black soil under rotary tillage (B-B),conventional no tillage (C-C),and no tillage and straw mulching with original crop (0-0) treatments.The results showed that no tillage with straw mulching significantly increased soil SOC,TN,and TP content compared to tillage;except for C/N,the C/P and N/P ratios of straw mulching in no tillage with straw mulching were higher than those in tillage.The soil SOC and TN content of rotary tillage and conventional no tillage were relatively reduced compared to tillage,while the C/N,C/N,and N:P values all significantly increased in the second year.Compared with tillage,the activities of four enzymes were significantly increased under the no tillage and straw mulching treatment.In the third year,rotary tillage significantly increased ACP and βG by 22.05% and 50.00%,respectively,compared with tillage.Rotary tillage,conventional no tillage,and no tillage with straw mulching all significantly increased soil enzyme C/P.The vector angles of soil enzyme activity under the tested cultivation methods were all less than 45 degrees,indicating that soil microorganisms in the experimental area may be limited by N;the length of enzyme vector increased significantly with age,indicating an increased degree of restriction by C.The results of principal component analysis,grey relational analysis,and correlation analysis showed that no tillage with straw mulching had the most significant impact on soil nutrient content and enzyme activity.In summary,the no tillage and straw mulching with original crop technology measures have a good improvement effect on activating soil nutrients and enzyme activity,as well as maintaining soil ecological stability.

The aim was to elucidate the effects of different foliar fertilizer nitrogen application amounts on nitrogen accumulation,translocation,and utilization in maize.This experiment was conducted in 2021—2022 using a randomized block design,with maize variety Lihe 1 as the research object.No fertilization treatment (CK),conventional root fertilization treatment (CF),foliar nitrogen reduction treatment of 20% (LF1),foliar conventional nitrogen application treatment (LF2),and foliar nitrogen increase treatment of 20% (LF3) were set up to analyze the differences in nitrogen accumulation,translocation,and utilization of maize under different nitrogen fertilizer application amounts,no fertilization and conventional root fertilization.The results showed that the nitrogen accumulation in maize stems and leaves showed a trend of first increasing and then decreasing with the advancement of the growth stage,reaching the maximum value at the tasseling and silking stage.The nitrogen accumulation per plant gradually increased with the advancement of the growth stage and reached its maximum value at mature stage,the nitrogen accumulation per plant was highest in the LF2 treatment.The proportion of nitrogen distribution in leaves was highest before the silking stage;after the silking stage,the proportion of nitrogen distribution in grains gradually increased,reaching its peak at mature stage.The CK had the lowest proportion of nitrogen accumulation in grains,while the LF1 treatment had the highest proportion of nitrogen distribution in grains in 2021,and the LF3 treatment had the highest proportion of nitrogen distribution in grains in 2022.The nitrogen transport rate and the contribution rate of nitrogen transport to grains first increased and then decreased with the increase of nitrogen application amounts,the nitrogen harvest index and nitrogen utilization efficiency decreased with the increase of nitrogen application amounts;in 2021 and 2022,the nitrogen utilization efficiency of LF1 treatment,LF2 treatment,and LF3 treatment was higher than that of CF treatment,and the nitrogen utilization efficiency of LF1 treatment was the highest.The nitrogen absorption efficiency of foliar nitrogen application treatment for two years was higher than that of CF treatment.There were no significant differences in ear length,ear thickness,and ear row number among the treatments.CK had the longest bald tip length,and the row number and hundred grain weight of each nitrogen application treatment were higher than those of CK.CF treatment had the highest biological yield,while LF1 treatment had the highest grain yield and harvest index.The grain yield of each treatment was significantly higher than CK,and the harvest index decreased with increasing nitrogen application amounts.Therefore,maize can achieve better growth effects under LF1 foliar nitrogen application in the central and western regions of Inner Mongolia.

To investigate the transcriptome differences between resistant and susceptible varieties of cauliflower after inoculation with Xanthomonas campestris pv.campestris (Xcc),and to identify genes associated with cauliflower resistance to black rot disease,the susceptible variety Y1-2 and the resistant variety EC-247 of cauliflower were selected as the research subjects.Total RNA was extracted from cauliflower leaves at 0,1,3,and 5 days post-inoculation with Xcc,respectively.High-throughput parametric transcriptome sequencing was then conducted utilizing the Illumina RNA-Seq platform,followed by Real-time Quantitative PCR for validation of selected differentially expressed genes(DEGs).DEGs associated with disease resistance were screened and analyzed.The findings revealed that 6 355 genes exhibited significant differential expression between resistant and susceptible cultivars across the four time points.KEGG enrichment analysis focused on plant disease resistance pathways,identifying 47 genes involved in plant-pathogen interactions and 61 genes related to plant hormone signaling.Cluster analysis of these gene expression levels disclosed specific genes,including one CDPK,four CMLs,one PTK,one CaM,one RLK,and one SGT1 in the plant-pathogen interaction pathway,and three auxin-responsive protein genes,a TIFY gene,an indole-3-acetic acid-amido synthetase gene,two brassinazole-resistant protein genes,and a Shaggy-associated protein kinase zeta gene in the plant hormone signaling pathway.Notably,the expression of these genes was significantly higher in resistant varieties compared to susceptible ones,indicating their active response to pathogen infection at various time points.The results indicated that these differential genes might be related to disease resistance in cauliflower,which provided important genetic resources and scientific basis for molecular breeding of disease resistance in cauliflower.

Preliminary transcriptomic analysis identified ZmRAV1 as a candidate gene involved in maize's response to drought stress. To further investigate its function, this study cloned the ZmRAV1 gene, conducted bioinformatics analysis of its coding sequence, and overexpressed this gene in Arabidopsis thaliana. The function of ZmRAV1 was validated by assessing the phenotypes and physiological and biochemical indices of the transgenic Arabidopsis lines under drought conditions. The results showed that the ZmRAV1 gene had a total length of 1 176 bp and encoded 389 amino acids.It had the highest proportion of irregular coils in its secondary structure and was a hydrophilic protein that did not contain signal peptides and was non transmembrane.Subcellular localization indicated that the protein was located in the nucleus.ZmRAV1 exhibited high conservation across different species.Phylogenetic analysis indicated that ZmRAV1 shares the closest evolutionary relationship with its homolog in Miscanthus sinensis, showing a high degree of homology. After drought stress treatment,the root length of Arabidopsis thaliana lines overexpressing ZmRAV1 during germination was significantly higher than that of wild-type (WT)lines.In the seedling stage,WT showed withering or even death after drought stress,while the survival rate was lower than that of overexpressing lines.Moreover,the POD and SOD activities of ZmRAV1 overexpressing lines were higher than those of WT after drought treatment,indicating that overexpression of ZmRAV1 gene could enhance Arabidopsis thaliana's resistance to drought stress.

The tail type of sheep is a complex trait formed by the interaction of genetic and environmental factors.circRNA is closely related to lipogenesis.To investigate the effect of circular RNA(circRNA)on the tail fat deposition of sheep,transcriptome sequencing and differential expression analysis of sheep tail fat were performed.Candidate circRNA associated with sheep tail fat were screened,and the regulatory network diagram of circRNA-miRNA-mRNA associated with sheep tail fat deposition was constructed,the selected circRNA were located,and their functions were verified.The results showed that a total of 679 differentially expressed circRNA were detected in the transcripts of adipose tissue of two different tail types of sheep,of which 422 were up-regulated and 257 down-regulated.Moreover,GO and KEGG functional enrichment analysis was performed on differentially circRNA target genes,which involved many biological development processes such as DNA metabolism,anatomical structure development,catabolic process,autophagy,carbohydrate absorption process,cell proliferation and lipid metabolism related to fat deposition.Target gene enrichment was involved in cell growth and apoptosis,cell motility,transport and catabolism,signal transduction,transcription and translation,amino acid anabolism and other functions,suggesting that these circRNA may participate in the deposition process of sheep tail fat through the above pathways.The selected differential circRNA_0018 was localized by fluorescence in situ hybridization and verified in the precursor adipocytes.The results showed that circRNA_0018 was a true and stable cytoplasmic ring molecule,and functional verification of circRNA_0018 showed that it could promote adipocyte differentiation.circRNA_0018 may be involved in the process of fat deposition and lipid metabolism in sheep.

To analyze the variability of the novel Goose astrovirus(GAstV)in Lu'an,Anhui Province and to express the VP27-VP34 fusion protein,the gout samples were collected from a farm in Lu'an.After confirming positivity via RT-PCR,the virus was isolated by passage culture in goose embryos.Then,the isolated strain was subjected to an animal regression test,whole genome amplification sequencing and genetic evolution analysis.Subsequently,the VP27-VP34 fusion protein of the isolated strain was induced and expressed,and purified recombinant protein was used to immunize 6-week-old female BALB/c mice to produce polyclonal antibodies.Serum antibody titers were assessed using agar diffusion methods,and the specificity of the polyclonal antibodies was detected by indirect immunofluorescence(IFA).The specificity of the antibody was detected by indirect immunofluorescence(IFA),and the titer of the prepared antibody was detected by the agar diffusion method.The results showed that one strain of GAstV,named AH-2021 strain,was isolated from clinical samples.The animal regression test showed obvious urate deposition on the surface of the heart and liver of goslings,and the kidney was white and swollen.Genetic evolution results revealed that AH-2021 belonged to GAstV-1,showing 98.0%—99.0% identity with other GAstV-1 strains in GenBank.The recombinant expression vector pCold-TF-VP27-VP34 was induced by IPTG to obtain the target protein,and SDS-PAGE showed that the molecular weight of the recombinant protein was about 110 ku,which mainly existed in the form of supernatant.IFA results showed that the polyclonal antibody was able to specifically recognize the GAstV,and the agar diffusion results showed that the titer of polyclonal antibody was up to 1:16.In conclusion,a strain of novel GAstV AH-2021 was isolated from gouty goslings,and animal regression tests showed that the novel Goose astrovirus was the pathogen causing gout in goslings,and a polyclonal antibody to the VP27-VP34 fusion protein was prepared.

To clarify the effects of different tillage methods on soil moisture dynamics changes and maize yield under the condition of straw return to field in the black soil area along the foothills of Daxing'anling,based on six consecutive years of tillage positioning experiments,this study analyzed the effects of seven types of tillage methods,namely,full-crushing and deep-tillage (SCD),full-crushing and shallow-tillage (SSS),full-crushing and deep-tillage (SCS),full-crushing and re-harrowing (SCR),full-crushing and rotary tillage (STR),full-crushing and no-tillage (NTS),and conventional tillage without returning straw to the field (CK)on soil moisture characteristics,water consumption,water use efficiency,and agronomic traits and yield of maize in the 0—60 cm soil layer at different growth stages of maize in each treatment.The results showed a bimodal pattern of soil quality moisture content in 2022 and 2023.0—10 cm soil layer soil quality moisture content was significantly higher than that of CK,and the NTS treatment had the highest soil quality moisture content in several periods.10—20 cm soil layer soil quality moisture content was lower than that of CK in the SSS and NTS treatments at the jointing stage,and soil quality moisture content was higher than that of CK in the 20—40 cm and 40—60 cm soil layers in 2022 and 2023 for all the treatments.In 2022 and 2023,maize plant height in all treatments except NTS treatment was significantly higher than CK at different growth stages.The SCD treatment was the tallest and the NTS treatment was the shortest at mature stage.Leaf area index (LAI) varied little among treatments at seedling stage of maize,STR treatment had the highest LAI after the jointing stage,and all treatments had significantly higher LAI than NTS treatment at the big trumpet stage.Dry matter accumulation in all treatments except for the SCS,NTS treatment was significantly higher than in CK,and dry matter accumulation was highest in the SCD treatment and lowest in the NTS treatment at mature stage.All tillage treatments increased maize yield and water use efficiency compared with CK,but the SCD treatment was significantly higher than CK.Comprehensive analysis of the indexes showed that the two tillage methods of full-crushing and deep-tillage and full-crushing and shallow-tillage were favorable to improve soil structure,maize yield and water use efficiency in the black soil area along the foothills of Daxing'anling.

In order to study the synergistic regulation characteristics of plant growth regulators on canopy-root traits of spring maize and to further reveal the lodging resistant mechanism of the plants,in 2021—2022,with the maize varieties Dika 159(DK159)and Xianyu 335(XY335)as experimental materials,at two planting densities of 75 000,90 000 plants/ha,plant growth regulator treatment (PGR) and water control (CK) were set up to analyze and compare the canopy structure,stem basel internode characters,root morphological characteristics and physicochemical properties of root bleeding sap under different treatments.The results showed that PGR had regulatory effects on both the maize canopy and roots.After PGR treatment,plant height,ear height and center of gravity height decreased,the mean leaf tilt angle above ear increased,light transmittance of ear layer increased by 23.59% on average,light transmittance of basel layer increased by 18.60% on average,and stem quality of basal internode was significantly improved.At the same time,the total root number,root length and root dry weight of PGR treatment increased,the root width at 10 cm below the surface increased,the root bleeding sap flow and nutrient flow increased,and the root morphological characteristics and transport capacity were obviously optimized.The flow of CTK and IAA in the root bleeding sap increased,while the flow of GA decreased.PGR effectively reduced the occurrence of stem lodging and root lodging through the synergistic regulation of canopy and root system,the field lodging rate of maize decreased from 13.43% to 6.47%,and the average yield of maize increased by 16.10%,thus achieving stable and high yield.

Soil nitrogen(N)cycling enzyme activity serves as a crucial indicator for characterizing soil fertility and N transformation.To investigate the effects of long-term application of fertilizers on the soil enzyme activities correlated with N cycling in rhizosphere soil of double-cropping rice fields in southern China,our project was based on a continue 37-year fertilization localization field experiment,including four fertilization treatments:without fertilizer as a control(CK),single fertilizer(MF),rice straw residue and mineral fertilizer(RF),and 30% organic manure and 70% mineral fertilizer(OM).The activities of N cycling enzymes in the rhizosphere soil were measured,and their correlation with soil chemical properties was analyzed.The results were as follows:compared to MF and CK treatments,OM and RF treatments significantly increased the contents of total N(TN),organic carbon(SOC),ammonium N($\mathrm{NH}_{4}^{+}-\mathrm{N}$),nitrate N($\mathrm{NO}_{3}^{-}-\mathrm{N}$)and microbial biomass N(SMBN)in rhizosphere soil,and also increased rice yield.The urease(Ure)and nitrite reductase(NiR)activities of rhizosphere soil in OM and RF treatments were significantly higher than those in MF and CK treatments.The RF treatment significantly increased rhizosphere soil hydroxylamine reductase(HyR)activities compared to the other three treatments,by 21.7%,13.0%,and 8.7%,respectively.This finding shown that OM treatment significantly increased protease(Pro),nitrogenase(Nit),nitrate reductase(NR)and nitrous oxide reductase(Nos)in rhizosphere soil compared to RF,MF and CK treatments.In comparison to MF treatment,OM treatments increased Pro,Nit,NR and Nos activities in rhizosphere soil by 20.0%,26.1%,426.1% and 26.7%,respectively.Nonetheless,the activity of nitric oxide reductase(Nor)on rhizosphere soil was considerably higher in the CK treatment than in MF,RF and OM treatments.Pearson correlation analysis revealed a substantial positive correlation between soil NR,NiR,Nit,Nos,Ure,Pro and soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN as well as rice yield.Soil Nor activity was observed to have a significantly negative connection with soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN and rice yield.The findings presented above showed that soil chemical properties and yield were substantially related to rhizosphere soil N cycling enzyme activities.Redundancy analysis(RDA)showed that the first order axis could explain 93.34% of the enzyme activity in rhizosphere soil and soil $\mathrm{NO}_{3}^{-}-\mathrm{N}$,TN and SOC contents were the key factors affecting the pattern of rhizosphere soil enzyme activities.Therefore,the long-term application of organic materials such as organic manure and rice straw can enhance soil chemical and biological characteristics,stimulate soil N cycling enzyme activities,and effectively fertilize paddy soils by partially replacing chemical fertilizers.

In order to provide production advice of high-quality upper tobacco leaves,field experiment was carried out to elucidate the effects of nitrogen application on the growth,development,carbon and nitrogen metabolism of upper leaves of different tobacco varieties.Three treatments,37.5(LN),75.0(MN)and 112.5 kg/ha nitrogen application rate(HN)were conducted with NC89 and Yunyan 87 as the test varieties.The effects of different treatments on the agronomic traits,photosynthetic characteristics,leaf tissue structure,key enzymes activity of carbon and nitrogen metabolism,and chemical composition of the upper leaves of were studied.The results showed that the leaf length,leaf width,leaf area,and leaf dry mass of the upper leaves of both varieties were significantly improved with the increased of nitrogen application rates.At 115 days after transplanting(DAT),the leaf area of NC89 and Yunyan 87 of HN were significantly increased by 63.10% and 68.43% compared to LN,respectively.Increasing nitrogen rate improved the leaf chlorophyll content of NC89,the leaf chlorophyll content of HN was 6.67%—37.50% higher than that of LN,respectively.The leaf net photosynthetic rate was notably improved by increasing nitrogen rate of Yunyan 87,especially at 70 DAT and 80 DAT.The leaf palisade tissue,leaf spongy tissue,and leaf thickness were markedly promoted with the increased nitrogen application rate on 85—115 DAT.The palisade tissue and spongy tissue thickness of LN and MN were almost stable from 95 to 115 DAT,but HN improved by 9.82%—14.08% and 10.72%—13.72% respectively.The leaf carbon content and leaf carbon and nitrogen ratio of both varieties were significantly decreased by reduced nitrogen rate,while leaf nitrogen content significantly increased.The activities of invertase,sucrose phosphate synthase,sucrose synthase,and glutamate synthase of both varieties were increased with the increased nitrogen application rate.HN decreased the amylase activity of Yunyan 87,but increased that of NC89.At 115 DAT,the amylase activity of HN of Yunyan 87 was 27.53% lower than MN,while HN increased amylase activity by 33.86% and 21.74% compared to LN and MN of NC89,respectively.The activity of glutamine synthetase of Yunyan 87 significantly increased with the increased nitrogen application rate,while there was no significant difference among different treatments of NC89.The content of flue-cured tobacco leaf reducing sugar and total sugar were reduced,but the content of nicotine and total nitrogen were improved with increased nitrogen application rate.Under the same nitrogen application rate,the content of flue-cured tobacco leaf nicotine,total nitrogen,and potassium of Yunyan 87 were higher than NC89,while the content of reducing sugar,content of total sugar(except LN),sugar and nicotine ratio,and nitrogen and nicotine ratio were lower than NC89.The effect of nitrogen application rate on upper leaves of tobacco varieties is difference.Increasing nitrogen application rate could promote the growth,development and carbon metabolism,reduce sugar and nicotine ratio and nitrogen and nicotine ratio,and improve the coordination of chemical components of NC89.However,increasing nitrogen application rate could lead to excessive nitrogen metabolism,delay the transformation from nitrogen metabolism to carbon accumulation metabolism,and cause late maturity of Yunyan 87.

To clarify the regulatory effect of nitrogen(N)fertilizer reduction and postponing on the productivity of wheat-maize double cropping system in Huang-Huai-Hai Plain.The annual N fertilizer experiment of summer maize and winter wheat was established of four N application systems:annual N fertilizer application 400 kg/ha of traditional farmer treatment(F₄₀₀),10% reduction of annual N fertilizer(FN),20% reduction of annual N fertilizer(FH),and 30% reduction of annual N fertilizer(FL)from 2020 to 2023 at Jiyang Experimental Base of the Shandong Academy of Agricultural Sciences in Jinan.The grain yield,aboveground N accumulation characteristics,N use efficiency,and the nitrate residue after harvest in the 0—200 cm soil layer of wheat-maize double cropping system were tested,in order to provide the theoretical basis for further optimization of N fertilizer management in Huang-Huai-Hai Plain.The results indicated that N fertilizer postponing was optimized the grain yield of summer maize and winter wheat under the condition of N reduction,and the averaged across the three years,FL significantly increased by 9.2%—18.1%,13.5%—20.5%,and 11.1%—19.1%,respectively,compared with F₄₀₀ and FN.N fertilizer postponing improved the N accumulation rate,and promoted aboveground N accumulation at wheat-maize different growth stages,and the averaged across the three years,FL significantly increased plant N accumulation by 5.7%—12.3% and 5.0%—12.8% under silking and maturity,respectively,compared with F₄₀₀,FN,and FH,as well as 8.2%—17.2% in grain N accumulation.For winter wheat,FL and FH treatments were significantly higher than F₄₀₀ and FN at jointing,anthesis,and maturity,and the averaged across the three years,FL and FH significantly increased by 23.4%—28.1%,20.7%—26.3%,and 12.6%—20.8%,respectively,compared with F₄₀₀,FN and FH,at the same time the grain N accumulation under FL significantly increased by 16.4%,15.0% and 5.8%,respectively,compared with F₄₀₀ and FN.N fertilizer postponing optimized the N use efficiency of wheat-maize double crop system,the averaged across the three years,FL significantly increased N uptake efficiency by 4.8%—57.7% and 32.0%—72.4% of summer maize and winter wheat,respectively,compared with F₄₀₀,FN,and FH;and FL significantly increased N partial factor productivity by 68.8% and 40.4% in summer maize,respectively,compared with F₄₀₀ and FN,as well as by 38.4%—71.8% in winter wheat compared with F₄₀₀,FN,and FH.At harvest of summer maize and winter wheat,the soil nitrate residue was mainly enrichment in the 0—40 cm soil layer under four N application systems,the averaged across the three years,accounted for 40.0%,38.9%,44.9%,42.5% and 37.3%,36.9%,46.7%,38.3% of the 0—200 cm soil layer,respectively.In addition,the obvious accumulated effects in 0—200 cm soil layer nitrate residue under F₄₀₀ and FN treatments at harvest of summer maize and winter wheat,but there was the relative balance was achieved under FL and FH treatments.In conclusion,a 30% reduction of annual N fertilizer by N fertilizer postponing could optimize plant N accumulation characteristics and realized synergistic improve grain yield and N use efficiency.Therefore,FL treatment was an optimal N application system for realizing the collaborative target of high-yield,high-efficiency,and environment-friendly of wheat-maize double cropping system in Huang-Huai-Hai Plain.

Light-harvesting chlorophyll a/b binding proteins are important in plant photosynthesis and abiotic stress response.To study the characteristics of GhLhcb2A1 and its expression patterns and functions in low temperature and drought response in upland cotton,full-length CDS of GhLhcb2A1 gene was cloned from the leaf cDNA of Jimian 262 by PCR.Bioinformatic analysis was conducted to learn the basic characteristics of the gene.The expression patterns and functions in low temperature and drought response were evaluated by qRT-PCR and virus-induced gene silencing.It was shown that the length of GhLhcb2A1 CDS was 798 bp,encoding 265 amino acids.GhLhcb2A1 was highly expressed in leaves and was significantly up-regulated in leaves and roots under low temperature and drought treatment.Compared with the control,its expression maximized at 3 h under low temperature and drought in leaves with 17.42 and 30.03 folds increase respectively,whereas maximized at 6 h under low temperature and 12 h under drought in roots with 11.65 and 65.04 folds respectively.Subcellular localization assay verified that GhLhcb2A1 was expressed in the chloroplasts of cells.Compared with the control plants,GhLhcb2A1 silenced plants showed a more severe phenotype of water loss and dryness under both low temperatures and drought.The accumulated malondialdehyde content in the leaves of the silenced plants was significantly higher than that of the control,while the proline content and superoxide dismutase activity were significantly lower than those of the controls,suggesting that GhLhcb2A1 silenced plant reduced the resistance to low temperature and drought.The above results implied that this gene played a positive role in regulating low temperature and drought response.

The aim was to clarify the effects of different nitrogen application levels on soil organic nitrogen fractions and nitrogen use efficiency in maize fields in the central-western region of Inner Mongolia,so as to provide a reference for the scientific management of soil nitrogen and sustainable development of modern agriculture.Six nitrogen application levels were set up,N0(0 kg/ha),N8(120 kg/ha),N12(180 kg/ha),N16(240 kg/ha),N20(300 kg/ha),and N24(360 kg/ha).The dynamic changes with effects of nitrogen application on soil total nitrogen content,particulate organic nitrogen content,light fraction organic nitrogen content and heavy organic nitrogen content,as well as maize yield and nitrogen use efficiency were analyzed at different soil layers at pre-sowing and post-harvest.The results showed that soil total nitrogen,particulate organic nitrogen,light fraction organic nitrogen,and heavy fraction organic nitrogen content decreased with deepening of the soil layer at the same nitrogen application level;soil total nitrogen content at pre-sowing increased with the nitrogen application levels.Soil total nitrogen content in the N16,N20,and N24 treatments was significantly higher than that in the N0,N8,and N12 treatments at post-harvest.Soil particulate organic nitrogen content of N16 treatment was highest in the 0—10 cm,10—20 cm,and 20—40 cm soil layers at pre-sowing,with 0.14,0.13,and 0.09 g/kg,respectively.At post-harvest,N16 treatment had the highest content in the 10—20 cm,20—40 cm,and 40—60 cm soil layers,with 0.19,0.10,and 0.09 g/kg,respectively.The highest increase of soil light fraction organic nitrogen content of 37.27% was in the N16 treatment,and the highest increase of soil heavy fraction organic nitrogen content of 7.35% was in the N24 treatment,followed by the N16 treatment,at 6.84%.The N16 treatment had the highest maize biological yield of 31 443.50 kg/ha;the highest maize economic yield of 18 526.47 kg/ha;and the nitrogen use efficiency decreased with the increase in nitrogen fertilizer application levels,N16 treatment in the nitrogen harvest index was the highest, at 79.20%.In conclusion,the more suitable nitrogen fertilizer application level in the central-western region of Inner Mongolia should be maintained under 240 kg/ha,in order to achieve the best soil nitrogen management ang crop yield.

In order to investigate the expression pattern and biological function of lncRNA TCONS_00143126 in E.coli type mastitis of cows in depth.This study used cDNA from bovine mammary epithelial cells as a template,and confirmed the presence of lncRNA TCONS_00143126 using PCR cloning and sequencing techniques.Subcellular localization analysis of lncRNA was performed,and potential target miRNAs and genes were predicted.The potential mechanism of its action in bovine mastitis was explored through KEGG pathway enrichment analysis.In addition,LPS was used to induce bMECs to construct an in vitro model of bovine mastitis,and the expression of lncRNA TCONS_00143126 in LPS-induced bMECs at 6,12 and 24 h was detected by RT-qPCR.The results showed that lncRNA TCONS_00143126 was real,and its expression was significantly up-regulated in LPS-induced bMECs,and it was mainly distributed in the nucleus.The results of target gene prediction and KEGG enrichment analysis showed that lncRNA TCONS_00143126 might regulate inflammatory signaling pathways such as JAK-STAT,mTOR and MAPK by targeting miRNAs(bta-miR-133a,bta-miR-193a-5p and bta-miR-375,etc.)and target genes(IFNE,SLC2A10,MEX3B),and then play a role in the inflammation of bovine mammary epithelial cells.

Anoctamin 5(ANO5)is a multichannel membrane protein localized in the sarcoplasmic and sarcoplasmic reticulum that primarily plays a role in myosin membrane repair and phospholipid scrambling,mutations in the ANO5 gene can lead to jaw hypoplasia as well as various myopathies.It aimed to investigate the association of SNPs in the ANO5 gene with fat deposition traits in sheep.A population of 1 005 healthy and clearly genealogical Hu sheep male lambs was selected for the study,and PCR amplification and KASPar typing techniques were used to detect the locus polymorphisms of the ANO5 gene in the experimental population and analyze the associations with fat deposition traits.The expression level of ANO5 gene in different tissues was analyzed by qPCR.The results showed that sheep ANO5 gene was widely expressed in a variety of tissues in Hu sheep,and the highest expression of ANO5 gene was found in heart tissue compared with other tissues.Three genotypes of CC,CT and TT with the g.58010 C>T polymorphic locus were detected in the 10th intron of the sheep ANO5 gene.Descriptive statistics showed that the perirenal fat weight was the most different and had the highest degree of variability compared with other fat weights.Correlation analysis showed that fat deposition related traits were positively correlated with growth and feed efficiency traits,and the results of association analyses showed that the polymorphic locus was significantly associated with perirenal fat weight and its related traits in the Hu sheep.Among them,the perirenal fat weight of individuals with CC genotype was significantly lower than that of individuals with TT genotype.In conclusion,the g.58010 C>T mutation locus of the ANO5 gene can be used as a candidate molecular marker for perirenal fat deposition traits in Hu sheep.

Soybean P34 protein mainly exists in soybean seeds,and its upstream promoter was likely to regulate the high expression of downstream genes in seeds.In order to further study the tissue expression pattern of soybean P34 protein gene and the regulatory activity of soybean P34 protein gene promoter,qRT-PCR was used to detect the expression of soybean P34 protein gene in soybean tissues.The 5'upstream sequence of soybean P34 protein gene(GmP34P)was cloned.The transcription initiation sites and cis-elements were analyzed by bioinformatics.The expression vector was constructed and the tobacco was transformed by Agrobacterium-mediated leaf disk method to detect GUS expression in transgenic tobacco.The results showed that the expression of P34 protein gene in soybean seeds was significantly higher than that in roots,stems,leaves and flowers.The length of GmP34P sequence obtained by cloning was 1 380 bp.Predictive analysis showed that the transcriptional start site of this sequence was base A at position 1 342,and the sequence contained a variety of cis-acting elements related to high seed expression,such as RY element,Skn-1 motif,2S seed protbanapa,etc.The plant expression vector pCAM-GmP34P containing GUS gene driven by GmP34P promoter was obtained.The positive transgenic plants were screened by hygromycin,PCR and RT-PCR.The results showed that GUS gene expression was extremely significant in transgenic tobacco seeds compared with other tissues by qRT-PCR with positive pCAM-GmP34P transgenic tobacco plants.GUS histochemical staining showed that the GmP34P promoter could regulate the high expression of downstream GUS gene in seeds.

As a key regulating factor in response to various abiotic stresses,plant heat shock transcription factor (Hsf) has a big family,and diverse structure,characteristics and functions.Hsf not only directly regulates Hsp and other relative gene expression and participates in the processes of response and adaption to various abiotic stresses,but also mediates many life activities regulation.Since the first Hsf was cloned from yeast in the 1980s,more and more Hsfs from other species have been identified and studied.In the previous reports,the identification of the Hsf family in plants was performed only in model species such as Arabidopsis and tomato.Furthermore,the studies is mainly focused on the HsfA subfamily,with few studies on the HsfB subfamily.And,the precise function of HsfC family is also largely unknown.With global climate change,the frequent occurrence of extremely high temperature events has seriously threatened the yield and quality of wheat,maize and other crops.To deal with the threat posed by heat stress,unraveling the mechanism of thermotolerance,identifying functional the targeted Hsfs and improving stress tolerance of crop through biotechnology methods is important.The number of Hsf family in field crops is various,the genome is complex,and the related research started lately compared with model species.To this end,our laboratory began to study the Hsf family of crops in 2009.Based on the latest genomic information,we confirmed the number of members,the modular structure and the spatio-temporal expression pattern of Hsf family.At the same time,with the help of transgenic wheat and mutant by genetic transformation and the CRISPR/Cas9 clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein mediated genome editing technology,several Hsfs were cloned and their regulatory functions of thermotolerance were identified,and some mechanism of thermotolerance was clarified.Our research not only enriched the theoretical basis of thermotolerance,but also provided new germplasm for biological breeding.At present,many studies have reported on functional identification and transcriptional regulation of Hsfs,however,evidences lack on which upstream component mediate Hsf's participation in regulation of thermotolerance,and the related mechanism is still unknown.Based on previous research results about wheat and maize Hsf families of lab,and many relative reports published in public,we reviewed the roles and mechanisms of plant Hsf in regulating process reported in recent years,aiming to promote research in illustrating the extensive and special roles and regulation network of plant Hsf family further,and dig useful genes and selective QTLs for biological breeding for plant thermotolerance.

Bioinformatics and expression patterns of Dicer-like(DCL),Argonaute(AGO)and RNA-dependent RNA polymerase(RDR)gene families in the whole genome of Solanum habrochaites were analyzed,so as to provide references for further study on the functions of DCL,AGO and RDR gene families in the response of S.habrochaites to abiotic and viral infection.Using Arabidopsis thaliana DCL,AGO and RDR genes as reference sequences,the genome of S.habrochaites LA1777 was searched by local perl language and software such as Pfam and SMART,and the members of ShDCL,ShAGO and ShRDR gene families were determined.Bioinformatics analysis of DCL,AGO and RDR family genes in S.habrochaites was carried out by means of ExPASy,GSDS 2.0,MEGA,Tbtools and SWISS-MODEL.According to abiotic stress treatment,Tomato chlorosis virus(ToCV)treatment and Real-time Fluorescence Quantitative PCR technology,the expression patterns of these genes were analyzed.Seven ShDCL,15 ShAGO and 6 ShRDR genes were identified from S.habrochaites,which were distributed on chromosome 5,7 and 6 respectively.The encoded proteins were similar in structure to DCL,AGO and RDR in other plants,and all of them contained conserved domains unique to this family.Phylogenetic analysis showed that these genes were divided into 4 subgroups,and there were high structural and functional similarities between S.habrochaites and S.lycopersicum.ShDCL2a,ShDCL2c,ShDCL3,ShDCL4,ShAGO1b,ShAGO3,ShAGO4b,ShAGO5,ShAGO7,ShAGO10a,ShAGO10b,ShRDR1,ShRDR2,ShRDR3a,ShRDR6a and ShRDR6b were significantly up-regulated after various abiotic stresses and ToCV infection.It is speculated that these genes play important roles in abiotic stress and virus infection.

In order to study the problem of strong winter/spring Brassica napus seed germination and flowering period under different winter sowing dates.Two strong winter rapeseeds and two spring rapeseeds provided by Gansu Agricultural University were used as materials.The experiment was carried out in the experimental field of Gansu Agricultural University from October 2022 to August 2023.The winter rapeseeds was carried out on October 11,2022.The winter/spring rapeseeds was sown every 20 days from December 10,2022,and the sowing ended on February 8,2023.The flowering period was recorded,and the germination seeds of winter rapeseed were sampled every 20 days to determine their physiological and biochemical characteristics and analyze the expression characteristics of vernalization genes(FLC,VRN2,FRI,FT).The results showed that the flowering period of winter/spring rape seeds was different by 22—34 days.The difference of flowering time between autumn sowing and spring sowing was 4—7 days.The flowering time of winter rapeseed in autumn sowing(October 11 th)was close to that of spring rapeseed under different winter sowing dates(December 10th,December 30th,January 19th,February 8th),and the flowering overlap time was as long as 15—20 days.With the delay of the sowing date,the relative expression levels of FLC,FRI and FT genes in germinating seeds of winter sowing were down-regulated.The relative expression of VRN2 gene was down-regulated in the early vernalization and up-regulated in the late vernalization.The activities of superoxide dismutase(SOD),peroxidase(POD),catalase(CAT)and the contents of soluble protein(SP),gibberellin(GA3)and salicylic acid(SA)in germinating seeds were increased in the early vernalization,but those were decreased in the late vernalization.The contents of malondialdehyde(MDA)and abscisic acid(ABA)were increased in rapeseed germinating with the increase of vernalization time.

This study explored the effects of intercropping and rotation on the growth,yield and quality of continuous cropping peanut,to provide theoretical basis for achieving high yield in peanut production.From 2022 to 2023,sweet potato-peanut rotation system(PSP)and maize-peanut intercropping and rotation system(PMP)were set up in the experimental farm of Henan University of Science and Technology on the basis of continuous cropping peanut for 2 years and 11 years respectively,with continuous cropping peanut as control(CCP1 and CCP2,respectively).The effects of PSP and PMP on photosynthetic characteristics,root characteristics,dry matter accumulation and distribution and yield of peanut were studied.The results showed that compared with CCP1,the leaf area index(LAI)of rotating peanut in PSP system(SRP)was significantly increased by 35.08%—53.68% and 24.32%—33.52% at pod-setting stage(PSS)and full pod maturity stage(PMS),respectively.The SPAD value at PSS and pod bulking stage(PBS)increased by 11.93%—18.55% and 5.95%—9.63%,respectively.Compared with CCP2,the LAI of rotating peanut in PMP system(MRP)increased by 46.81%—57.96% and 27.00%—61.78% at PSS and PMS,respectively.At PSS and PBS,compared with CCP2,the SPAD value of MRP and intercropping peanut(MIP)increased by 3.32%—3.69%,7.50%—8.64% and 5.47%—18.37%,15.73%—31.11%,respectively.At PSS and PBS,compared with CCP1,the net photosynthetic rate of SRP increased by 23.68%—41.31% and 26.52%—32.55%,and compared with CCP2,MRP increased by 12.77%—17.81% and 16.88%—62.07%,respectively.They both significantly improved the root length and root tip number,and promoted the dry matter accumulation and the distribution to pods during PMS,and the yields increased by 31.42%—47.36% and 54.12%—75.09%,respectively.Compared with CCP2,MIP reduced the LAI,net photosynthetic rate,root length,root tip number,as well as dry matter accumulation and yield of peanut under the influence of maize shading.At the same time,the content of peanut oleic acid and oleic acid-linoleic acid ratio was significantly increased after rotation.Among them,SRP increased by 1.63—1.65 percentage point and 6.59%—10.52%,respectively,compared with CCP1,and MRP increased by 1.95—2.82 percentage point and 9.75%—14.16% compared with CCP2,respectively.In summary,sweet potato-peanut rotation and maize-peanut rotation increased the peanut yield compared with continuous cropping peanut,the reason was that sweet potato-peanut and maize-peanut rotation promoted peanut root growth,delayed the leaf senescence,and increased photosynthetic rate,especially the photosynthetic rate during late growth period,which promoted the dry matter accumulation and distribution to seeds.Besides that,they could improve the quality of peanut to a certain extent.

This study systematically investigates the effects of silicon-modified biochar (MSC) on the chemical properties of acidic soil,organic carbon and silicon fractions,and the growth of tomato plants.Silicon-modified biochar was prepared,with a focus on investigating its impacts on carbon and silicon chemical fractions,and the availability in acidic soils;tomato growth and soil microbial activity were also evaluated.The results showed that silicon-modified biochar significantly increased soil pH,cation exchange capacity,electrical conductivity,available phosphorus and potassium.MSC also raised the levels of water-soluble sodium and iron in the soil and enhanced the activities of hydrogen peroxidase and sucrase enzymes,thereby improving soil quality.Both biochar modification and unmodified biochar significantly increased the content of different carbon fractions in the soil.Compared with unmodified biochar,silicon-modified biochar significantly increased soil microbial biomass carbon(21.9%) and water-soluble organic carbon (898.3%).Furthermore,silicon-modified biochar significantly increased the contents of soil available silicon,water-soluble silicon,free silicon,active silicon,iron-manganese-bound silicon and amorphous silicon by 362.6%,158.9%,18.1%,34.9%,193.8%,and 74.1%,respectively.Meanwhile,the application of biochar promoted the growth of tomato plants and the absorption of silicon nutrients,with modified biochar showing more pronounced effects.The accumulation of plant dry matter,silicon content,and absorption rate increased by 82.0%,98.9%,and 261.5%,respectively.In summary,silicon-modified biochar significantly affected the carbon and silicon chemical forms and transformation in the soil,increased soil effectiveness and enzyme activity,thereby promoting nutrient absorption and growth of crops,demonstrating its good potential application in agricultural production.

To study the photosynthetic characteristics,fluorescence parameters and yield response to phosphorus in maize,and to clarify the optimal phosphorus application rate for maize under drip irrigation and water fertilization technology.Providing solid theoretical basis and technical support for high-yield and high-efficiency cultivation of maize in Ningxia region.The experiment was carried out at Pingjipu Farm,Yinchuan,Ningxia,from 2019 to 2020,with six phosphorus treatments in the order of 0(P0),60(P1),120(P2),180(P3),240(P4),and 300 kg/ha(P5).Analysis of the changing patterns of photosynthetic and fluorescence parameters of spring maize leaves and their correlation with yield under different phosphorus fertilizer treatments.In two years,during the big bell mouth stage,leaf area index (LAI) was increased by 4.21% to 12.78% and 4.68% to 15.60% for P3 compared to other treatments,respectively.Phosphorus fertilizer at 180 kg/ha was most effective in promoting leaf area index and photosynthetic potential(LAD) of maize.LAD was significantly increased by 14.42% under P3 treatment compared to no phosphorus fertilizer treatment during the full two year period.The photosynthetic characteristics of maize responded differently to the intensity of phosphorus application,and as the intensity of phosphorus application increased,the net photosynthetic rate (Pn) all reached the maximum value after the stamen pumping stage,and at the R1 stage of the 2 years,the Pn was significantly increased by 10.68% under the P3 treatment as compared to the no-phosphorus-fertilizer treatment.The coefficient of determination (R²) was 0.926 5,0.889 9,and 0.832 0,respectively.Phosphorus application increased the maize photosystem Ⅱ composite performance index (PI),which had its maximum peak at the R1 stage in 2 years,and PI increased by 1.12% to 8.50% and 8.47% to 15.40% under the P3 treatment compared with the other treatments,respectively.The maximum yield was obtained at 180 kg/ha of phosphorus application,which was 17.27% higher as compared to no phosphorus treatment.Based on the analysis of the yield fitting equation,it was shown that the maximum corn yield of 13 823.84 kg/ha was reached at 179.34 kg/ha of phosphorus applied.Pearson's correlation analysis showed that appropriate leaf area index significantly affected maize yield in the late stage,and the photosynthetic parameters all had highly significant effects on maize yield completion;principal component analysis showed that the P3 treatment had the highest composite score for the optimization effect on maize yield.Reasonable transportation of phosphorus fertilizer can effectively ensure higher SPAD value,PSⅡ reaction center activity,improve the capture and utilization of light energy in spring maize,and promote photosynthesis,so as to improve the yield and economic benefits of maize.

In order to clarify the role of the Zn(Ⅱ)2Cys6 transcription factor gene VDAG_ 04814 in the growth, development and pathogenicity of the Verticillium dahliae. It constructed a VDAG_04814 gene knockout mutant using homologous recombination mediated by polyethylene glycol. Wild-type and mutant strains were inoculated separately onto PDA media supplemented with hydrogen peroxide, sodium chloride, potassium chloride, sorbitol, sodium dodecyl sulfate, Congo red, as well as onto media overlaid with sterile cellophane, to analyze their levels of resistance to oxidative stress, salt stress, osmotic stress, stress on cell wall and plasma membrane integrity, and strain penetration ability. Their pathogenicity was assayed, and the fungal biomass in potato plants was detected. After hygromycin selection and PCR validation, the correct knockout transformants were able to amplify DNA bands of 1 500 bp upstream and downstream, respectively, as well as the full-length 4 500 bp knockout fragment sequence. The results demonstrated that the growth rate and melanin formation ability of the ΔVDAG_04814 mutants were significantly reduced. On media subjected to oxidative stress and salt stress with the addition of hydrogen peroxide, sodium chloride, and potassium chloride, ΔVDAG_04814 showed a higher inhibition rate compared to the wild-type. On osmotic stress media with sorbitol, the growth inhibition rate of ΔVDAG_04814 was significantly lower than the wild type. No growth inhibition was observed for ΔVDAG_04814 on media subjected to cell wall and membrane integrity stress with the addition of sodium dodecyl sulfate and Congo red. On media overlaid with sterile cellophane, no colonies grew for ΔVDAG_04814, whereas the wild-type strain produced normal colonies. Pathogenicity tests indicated that the wilting index of ΔVDAG_04814 was significantly reduced compared to the wild-type, with wilting index ranging from 47.22 to 55.56. It has been demonstrated that VDAG_04814 can regulate the growth, development, stress resistance, penetration ability and pathogenicity of V. dahliae towards potato. This study provides a new target for the control of potato Verticillium wilt disease.

Investigating the predominant genotypes of Bovine viral diarrhea virus(BVDV)infecting cattle in Tongliao,Inner Mongolia,to provide reference for the BVDV epidemiology and prevention and control.In the preliminary phase of the experiment,fecal samples from diarrheic calves were collected at a cattle farm in Tongliao,Inner Mongolia.These samples were tested using PCR to detect BVDV positivity.Positive fecal samples were then inoculated into madin-darby bovine kidney cells(MDBK)for isolation.The isolated strains were identified using RT-PCR and indirect immunofluorescence staining.Subsequently,the full-length genome of the isolates was sequenced,followed by genetic evolution analysis and genotype determination based on sequences of the 5'UTR,N^pro,and E2 genes.The results indicated that this experiment successfully isolated a strain of BVDV,designated as NM-21.Inoculation of NM-21 into MDBK did not induce cytopathic effects,indicating it was a non-cytopathic strain(NCP).Both RT-PCR and indirect immunofluorescence staining confirmed its positivity,with a virus titer of 10^-3 TCID₅₀/mL.Based on the full-length genomic sequence,and homology and genetic evolution analysis of the 5'UTR,N^pro,and E2 gene sequences,the isolate NM-21 showed the highest nucleotide homology with the BVDV-1c subtype strain NM2103(GenBank accession number ON337882.1)from Inner Mongolia,China.

The highly contagious gastrointestinal infectious disease caused by Porcine epidemic diarrhea virus(PEDV)has led to significant economic losses in China's pig industry.It aimed to establish a basis for PEDV antibody detection methods and functional research of the N protein through the screening of specific nanobodies(Nbs)using phage display technology.Peripheral blood lymphocytes were isolated from a camel immunized with the N protein,and total RNA was extracted and reverse transcribed into cDNA.The variable domain of the heavy chain of heavy chain antibodies(VHH)was amplified by PCR,subcloned into the pCANTAB5E-ccdb vector,and electroporated into ER2738 competent cells to construct the VHH phage antibody display library.Subsequently,the library was subjected to four rounds of panning against the PEDV N protein,and positive phage clones were cloned into the pET-30a vector.The binding affinity and specificity of the Nbs were determined by indirect ELISA and Western Blot.The results showed that after the fifth immunization,the antibody titer reached 1∶25 600.The constructed phage display library had a capacity of 4.72×10⁸ and an abundance of 4.3×10¹⁰ cfu/mL,with a 93.75% positive rate.After four rounds of screening,16 Nb clones with different amino acid sequences were obtained,and Nb45 was validated to possess excellent specificity and binding ability to the PEDV N protein.This study successfully screened and obtained specific N protein-targeting Nbs,providing biological materials for the establishment of PEDV detection methods and foundational research.

In order to investigate the function of watermelon calcium-dependent protein kinase (CDPK) in grafted seedlings and abiotic stress environments, this study used RT-PCR technology to clone the ClCDPK(Cla97C01G019720) gene from watermelon grafted seedlings and performed bioinformatics analysis on it. Further designed specific primers with Kpn Ⅰ and Sal Ⅰ enzyme cleavage sites based on the ClCDPK sequence,conducted amplification and double enzyme cleavage, and connected with pCAMBIA1300 to successfully construct the expression vector pCAMBIA1300-35S-ClCDPK for the target gene.Using RT-qPCR technology, the gene expression levels of ClCDPK were measured in self rooted seedlings (ZG) and grafted seedlings (JJ) after being subjected to salt and drought stress, respectively.The results showed that the ORF of ClCDPK gene was 1 647 bp, encoding 548 amino acids. Its protein contained STKc_CAMK and FRQ1 functional domains, and was a hydrophilic protein. Subcellular localization prediction showed that the protein was located in the nucleus. Evolutionary tree analysis of ClCDPK with CDPK from six other plants revealed that it was closely related to CDPK from Cucurbitaceae melons and pumpkins, with protein sequence homology alignment exceeding 92.64%, indicating high homology.The RT-qPCR expression results showed that the expression level of ClCDPK in grafted seedlings was significantly higher than that in self rooted seedlings. With the duration of stress, the expression levels of ClCDPK in grafted and self rooted seedlings first increased and then decreased, and under the same stress treatment, the expression level of ClCDPK in grafted seedlings was higher than that in self rooted seedlings.This study indicated that ClCDPK responded positively to salt and drought stress, and the ability of grafted seedlings to resist stress was higher than that of self rooted seedlings. It is speculated that ClCDPK is one of the key factors in watermelon's response to grafting, thereby improving the salt and drought resistance of watermelon grafted seedlings.

To investigate the role of calcium-dependent protein kinase (CDPK) in wheat growth and stress response,the TaCDPK17 gene was cloned from common wheat and its sequence structure,expression pattern,and stress resistance function were preliminarily analyzed.The results showed that the length of the TaCDPK17 gene coding region was 1 701 bp, encoding 566 amino acids and possessing typical structural features of the CDPK family, including one conserved serine/threonine kinase domain and four EF hand shaped domains. Evolutionary tree analysis of TaCDPK17 and CDPK17 from 12 other plants showed that TaCDPK17 had high homology with the CDPK17 sequence of gramineous crops,especially Aegilops tauschii and barley.The promoter region of TaCDPK17 gene contained multiple cis regulatory elements related to hormone signaling pathways,light response.Among them, there are more abscisic acid (ABA) responsive elements (ABRE) and methyl jasmonate responsive elements (CGTCA). The expression analysis based on Real-time Fluorescence Quantitative PCR showed that the expression level of TaCDPK17 increased to varying degrees after induced by 100 μmol/L ABA, 100 μmol/L methyl jasmonate, 20% PEG6000, and 250 mmol/L NaCl. Under stress conditions of 2 μmol/L ABA and 100 mmol/L NaCl, the germination rate of Arabidopsis seeds overexpressing TaCDPK17 was significantly higher than that of the wild type. Meanwhile, overexpression of TaCDPK17 alleviated the inhibitory effects of ABA or osmotic stress treatments on seedling root growth. During stomatal closure, transgenic plants overexpressing TaCDPK17 are more sensitive to ABA and exhibit a stronger stomatal closure trend compared to wild-type plants. These results indicated that TaCDPK17 plays an important role in stress response and hormone signaling in wheat.

In order to explore the effects of different irrigation methods and nitrogen fertilizer management on late rice growth,development,yield formation,and nitrogen use efficiency,Y Liangyou 911 was used as test material.Two irrigation methods(W1.flooded irrigation;W2.moistening irrigation) and three nitrogen fertilizer managenments base,tiller,ear and grain fertilizer ratios(N1:5∶3∶2,N2:3∶4∶3 and N3:3∶4∶2∶1) were designed.No fertilization served as the control (CK1.flooded irrigation;CK2.moistening irrigation).Leaf area index,SPAD value of rice leaves,dry matter mass,yield formation and nitrogen use efficiency were determined under each treatment combination.The results showed that compared with W1,the LAI of rice treated with W2 was lower in the early growth stage and higher in the middle and late growth stages.The SPAD value of W1 treatment was higher,but there was no significant difference in SPAD value in the late growth period.Under the same irrigation conditions,compared with N1,N2 and N3 treatments it could delay the decline of LAI and SPAD values in the late growth period of rice.W2 treatment could significantly increase rice dry matter accumulation by 6.61%-16.37% compared with W1 treatment.Under nitrogen fertilizer treatment,the dry matter mass was higher in the early and middle stages of growth with N1,and the dry matter mass was higher in the later stages with N1 and N3.The yield increase of W2 mode was 7.59%-10.47% compared with W1 mode.The yield of W2N3 treatment was 3.24%-14.53% higher than that of other treatment.Although the effective panicle number was lower in W2N3 treatment,other yield components were increased,which resulted in higher yield.During the two years,W2N2 and W2N3 had higher values of total nitrogen accumulation and nitrogen absorption efficiency,W2N3 had higher values of nitrogen agronomic utilization,nitrogen partial productivity and nitrogen harvest index,and W1N3 had the higher value of physiological nitrogen utilization.In conclusion,irrigation methods and nitrogen management significantly affect rice yield and nitrogen uptake and utilization.The W2 (moistening irrigation) coupled with N3 (base fertilizer∶tillering fertilizer∶ear fertilizer∶grain fertilizer=3∶4∶2∶1) nitrogen management method is more conducive to rice dry matter accumulation,yield improvement and efficient use of nitrogen fertilizer,which can not only meet high yield,but also play a role in water saving.It is the best coupling method of water and fertilizer.

In order to investigate the effects of density on individual and population structure characteristics and yield of winter wheat under rainfed and limited water supply conditions,a field experiment was carried out at Gaocheng Experimental Station,Shijiazhuang City during 2022-2023 season using JM22 wheat cultivar under four densities:which were 1.8×10⁶ (D180),3.0×10⁶ (D300),4.2×10⁶ (D420) and 5.4×10⁶ (D540) plants/ha.Two irrigation treatments for each density,which were no irrigation during the whole growth period (W0) and irrigated once at the jointing stage (W1).The influence of planting density and irrigation treatments on leaf area (flag leaf,top 2 nd leaf,top 3 nd leaf,top 4 th leaf),non-leaf green organs (ear,awns,stem sheath) area,leaf area index,non-leaf green organ area index,dry matter accumulation,photosynthetic active radiation interception rate,water use efficiency (WUE) and yield of winter wheat were studied.The results showed that the leaf area of each leaf layer and non-leaf green organs decreased with the increase of density.D300 treatment got the highest leaf area index and non-leaf green organs index,and significantly higher than that in D540.The contribution of post-anthesis dry matter to grain yield was more than 70%.Compared with D540 treatment,reducing plant density decreased the transfer of dry matter before anthesis,but increased the accumulation of dry matter after anthesis and its contribution to grain yield.With the increase of density,WUE increased first and then decreased,D300 treatment achieved the highest WUE.Specifically,the WUE of D300 was 1.2%-14.4% and 2.5%-12.7% higher than that of other densities under W0 and W1,respectively.Compared with W0,W1 treatment increased the area of leaf and non-leaf green organs of different densities,delayed leaf senescence,and improved the photosynthetic active radiation interception rate of canopy.Ultimately,the grain yield increased by 28.1%-39.7%.Under the conditions of this experiment,D300 treatment increased the leaf and non-leaf green organ area,leaf and non-leaf green organ area index,canopy photosynthetically active radiation interception rate,post-anthesis dry matter accumulation and its contribution to final grain yield.The yield of D300 was 2.4%-6.6% and 0.3%-9.7% higher than that of the other densities under W0 and W1,respectively,which was the optimal density in this study.

The adult plant resistance gene Lr12 exhibits excellent resistance in production systems.To fine map and develop reliable molecular markers for Lr12,a cross was made between the susceptible variety Thatcher and the resistant near-isogenic line RL6011 containing the Lr12 gene.The F₁ generation resulting from this cross was self-pollinated to generate F₂ individual plants and F_2∶3 families.Field evaluations were conducted using a mixture of five highly virulent leaf rust pathotypes (PHTT, THKS, THTT, PHTS, and PHKS) to inoculate F₂ individual plants and F_2∶3 families for adult plant resistance assessment and genetic analysis of resistance.Subsequently,genotyping was performed using a 16K liquid chip on 10 resistant and 10 susceptible individuals from the F₂ generation to identify SNP markers closely linked to Lr12.This enabled the determination of the chromosomal physical interval containing the resistance gene,the development of SSR molecular markers,and the construction of a genetic linkage map.The results indicate that the segregation ratio of resistance to leaf rust in 3 494 F₂ individuals derived from the RL6011(Lr12)/Thatcher cross was consistent with a 3∶1 ratio ( {\mathrm{\chi }}_{3:1}^2=0.14;P=0.71). In the assessment of 685 F_2∶3 families, the segregation ratio among resistant individuals, resistant heterozygous individuals, and susceptible individuals conformed to a 1∶2∶1 ratio ( {\mathrm{\chi }}_{1:2:1}^2=2.01;P=0.37), suggesting that Lr12 is a dominant gene and the population segregation follows Mendelian single-gene inheritance patterns. Genetic linkage map analysis localized the adult plant leaf rust resistance gene Lr12 between SSR molecular markers YK12817 and YK12928,within a genetic interval of 0.38 cM.This corresponds to a physical interval of 2.09 Mb within the physical range of 579.44 Mb to 581.53 Mb on chromosome 4BL of the Chinese Spring reference genome(IWGSC.Ref.V1.0).These findings provide a solid basis for predicting candidate genes.

In order to analyze the possible role of dTLR2 in duck innate immunity,we study the expression of dTLR2 mRNA in immune organs during 1 to 10 weeks,as well as the expression in blood after challenging with Escherichia coli and Salmonella enteritis by Fluorescence Quantitative PCR.We also prepared polyclonal antibodies of the extracellular domain of dTLR2 by recombinant protein expression for further studying the biological function of dTLR2.The results showed that dTLR2 mRNA was expressed in spleen,thymus and bursa at different weeks.The expression of dTLR2 mRNA was significantly different among different weeks in the spleen and thymus tissues,and there was no significant difference in bursa.After infection with E.coli and S.enteritis,the expression of dTLR2 mRNA was significantly higher than that of the control group in the first and second days,but there was no significant difference among groups in the third day.Analyzing the sequence of dTLR2 gene,we predicted the extracellular domain,designed primers for PCR assay,and built pET28a-TLR2 recombinant plasmid.The recombinant plasmid was translated into E.coli BL21 (DE3) for expression of dTLR2-His protein.The SDS-PAGE results showed that the fusion recombinant protein efficiently expressed in the supernatant,molecular mass was around 29 ku.The results of Western Blot showed that the antibody which harvested from immunizing rabbits could detect recombinant dTLR2 and endogenous dTLR2.The analysis of the expression of dTLR2 and successful preparation of polyclonal antibody provide favorable support for further studying of dTLR2 biology function.