Glyphosate is currently the most widely used broad-spectrum herbicide.Cultivating glyphosate tolerant crops will help improve the effectiveness of chemical control on weed in farmlands,reduce the use of pesticide,and simplify preventive and control measures.To fully detect the glyphosate tolerance(GT)loci in wheat,484 germplasm resources from the Huang-huai wheat region were used to identify glyphosate toxicity.Based on the wheat 15K SNP array data,genome-wide association analysis(GWAS)was used to explore QTL related to glyphosate tolerance in wheat.The main results were as follows:the trend of changes in glyphosate tolerance of wheat varieties cultivated in different eras was slow,and the glyphosate tolerance had not significantly improved;three glyphosate tolerant wheat germplasms(including Henong 130,Jimai 782 and Taishan 23)were selected based on the phenotypic identification results of pesticide damage;seven QTL associated with the level of wheat pesticide damage were detected by GWAS,including 19 significant SNPs,distributed on wheat chromosomes 1A(0.00—30.48 Mb),1B(6.57—30.57 Mb),1D(0.00—22.98 Mb),4A(656.09—680.09 Mb),5A(508.19—532.19 Mb),6A(54.56—85.09 Mb),and 6D(12.02—36.02 Mb);the two QTL qGlyT-1A and qGlyT-6A located on wheat chromosomes 1A and 6A were the main effector sites for glyphosate tolerance in wheat,containing a total of 16 genes that may be related to glyphosate tolerance in wheat.

WRKY transcription factor is a family of transcription factors unique to plants.Studies have demonstrated that WRKY transcription factor played an important role in plant growth and development and in plant response to biological and abiotic stress.In order to reveal the function of tomato WRKY gene,two inbred lines of tomato with high resistance to bacterial wilt Hm 2-2(R)and high susceptibility to bacterial wilt BY 1-2(S)were used as experimental materials based on the preliminary transcriptome data,and a WRKY transcription factor SlWRKY75 gene(Solyc05g015850.3)was cloned.The structure,expression pattern and function of the gene and its encoded protein were analyzed by bioinformatics analysis,multiple alignment of amino acid sequences,phylogenetic tree construction,Real-time Quantitative PCR(qRT-PCR)and virus induced gene silencing(VIGS).The results showed that the full length of the cDNA of this gene was 653 bp,its maximum open reading frame was 519 bp,encoding 172 amino acids,the relative molecular weight of the protein was 19.878 51 ku,the theoretical isoelectric point was 9.32.The protein belonged to the hydrophilic non-secreted protein,and there was no transmembrane structure.Meanwhile,the protein had a highly conserved WRKY domain and a CX₄CX₂₃HXH zinc finger motif,which belonged to the Class Ⅱ family.Phylogenetic tree analysis showed that SlWRKY75 was closely related to Solanum pennellii SpWRKY75 and clustered into a group with other Solanaceae,while it was far related to Hevea brasiliensis HbWRKY75 and Gossypium hirsutum GhWRKY75 and was in different branches in the phylogenetic tree.The results of qRT-PCR analysis showed that the expression of SlWRKY75 gene were tissue-specific and could be induced by Ralstonia solanacearum,salicylic acid and jasmonic acid.VIGS result showed that silencing SlWRKY75 reduced plant resistance to bacterial wilt,indicating that SlWRKY75 positively regulated tomato resistance to bacterial wilt.These results suggested that SlWRKY75 gene played an important role in regulating tomato resistance to bacterial wilt.

In order to explore the gene expression regulation mechanism in the formation of reactive oxygen species (ROS) induced by heat stress,heat stress treatments were set at seedling,heading and filling stage to investigate the dynamic change of ROS accumulation in rice,respectively.And the Quantitative Real-time PCR (qRT-PCR) was used to analyze the expression pattern of nine respiratory burst oxidase homologue (Rboh) encoding genes (OsRboh1-OsRboh9) in rice under different growth stages.Results showed that the ROS accumulation in rice leaves and grains significantly increased with the extension of heat stress.The ROS content increased slowly after 7 days of heat stress at seedling stage, while increased continuously after heat stress during the heading stage and early filling stage (1-10 d) in rice grains. Expression levels of the nine OsRboh family genes continuously increased with the extension of heat stress at seedling and heading stages,and OsRboh7 and OsRboh5 showed higher expression levels under heat stress.The expression levels of OsRboh1,OsRboh5 and OsRboh9 were continuously increased,while other genes showed a change tendency of increasing initially and then decreasing under heat stress at filling stage.The expression levels of OsRboh7 and OsRboh5 were all reached to a high level at seedling,heading and filling stage under heat stress.Furthermore,higher expression levels of OsRboh7 and OsRboh5 were showed in various tissues and organs of rice such as seedling leaves,flag leaf,floret,lemma,palea,stamen,pistil and grain.The higher induced range of gene expression levels in OsRboh7 and OsRboh5 by heat stress was shown in seedling leaves,floret,stamen,pistil and grain.Taken together,OsRboh7 and OsRboh5 were mostly remarkably responsive to heat stress at different growth stages among the OsRboh family genes in rice,which indicated that OsRboh7 and OsRboh5 played a vital role in the regulation of ROS formation pathway in rice under heat stress condition.

Auxin response factor(ARF)is a class of transcription factors with B3 domain,which is a direct molecule regulating auxin response and controlling gene expression.A gene,ZmARF10,which encoded ARF protein and actively participated in drought-rehydration stress response,was previously screened in maize by analyzed transcriptome data.To further research the molecular mechanism of ZmARF10 regulating drought resistance of maize,and also provide a new idea for molecular breeding of drought resistance,the gene was firstly analyzed by bioinformatics software.Secondly,Quantitative Real-time polymerase chain reaction(qRT-PCR)was used to detect the expression pattern of ZmARF10 in different tissues,under high temperature,drought,high salt,ABA and restoring treatments,and in different inbred lines.Finally,the function of ZmARF10 was analyzed using CRISPR/Cas9 technology.The results showed that ZmARF10 was located on chromosome 3 of maize,with a total length of 2 127 bp,and encoded 708 amino acids with a typical B3 domain.The upstream 2 kb region of ATG of this gene contained response elements related to methyl jasmonate,auxin,abscisic acid and low temperature.The phylogenetic tree showed that the protein encoded by the ZmARF10 gene was closely related to sorghum.qRT-PCR results showed that ZmARF10 was a constitutive expression gene,and the expression level of ZmARF10 was the highest in mature corn roots of maize.Under high temperature,drought,high salt and ABA treatments,the expression of this gene was significantly up-regulated,and the up-regulation ratio was up to 8.2 times after drought stress.After drought stress,the expression level of ZmARF10 gene was significantly higher in the drought-resistant inbred line Zheng 36 than that in the drought-sensitive inbred line B73.Investigation of Arabidopsis wild type and ARF10-deficient mutants showed that,compared with wild type,the mutant plants showed leaf wilting and even dry death,roots curled,root branch number decreased,and lateral root growth and development were hindered under drought conditions.Determination of physiological and biochemical indexes showed that the relative water content,chlorophyll content and net photosynthetic rate of the deficient mutants were significantly lower than those of the wild-type plants after drought stress,indicating that the drought resistance of Arabidopsis decreased after ARF10 gene was knocked out.

In order to analyze the molecular mechanism of the formation of pink stripes on the petals of Dayuanyangjin,a Rhododendron cultivar,and to explore the key genes related to the formation of pink stripes,the petals of Dayuanyangjin in its full bloom period were selected to determine the pigments in the white and pink stripe regions of the petals,and at the same time,their transcriptome was sequenced and analyzed using Illumina HiSeq^TM sequencing platform.Sequencing results were verified by Real-time Fluorescent Quantitative PCR (qRT-PCR).The results showed that the formation of pink stripes on the petals of Dayuanyangjin was caused by the accumulation of anthocyanins.A total of 7 086 differentially expressed genes (DEGs) were identified in the pink striped and white regions of petals,of which 3 802 were up-regulated and 3 284 were down-regulated.The anthocyanin biosynthesis,phenylpropanoid biosynthesis and flavonoid biosynthesis pathways related to anthocyanin formation were more active in the pink stripe region of petals.According to the results of the GO and KEGG enrichment analysis of DEGs,31 DEGs were screened from the related and regulatory pathways of anthocyanin biosynthesis,of which 26 were structural genes of anthocyanin biosynthesis pathway and 5 were transcriptional regulation genes (3 MYB and 2 bHLH);26 anthocyanin anabolic structure genes encode 9 enzymes,and the expression levels of 20 genes in the pink stripe tissue of petals were significantly higher than those in the white petal tissue.Through NCBI homology search,it was found that two R2R3-MYB and one bHLH were similar to the known homologous genes related to the regulation of anthocyanin synthesis in fruits or leaves,and their expression levels in the pink stripe region of petals were also significantly higher than those in the white region.Ten differentially expressed genes related to anthocyanin biosynthesis were selected and verified by qRT-PCR.

The identification of leaf shape mutant and genes controlling leaf shape can not only provide germplasm resources for genetic improvement of leaf shape,but also help to analyze the genetic regulation mechanism of leaf development.vrnl11 was identified from an EMS induced Wankelü 3(WK3)mutant library.Progeny populations derived from vrnl11/WK3 and vrnl11/Zhonglü 1 were used for genetic analysis,and the segregation pattern of different phenotypic plants in F₂ populations was determined by χ² test.Two F₂ populations constructed by crossing vrnl11 with Zhonglü 1 and Zhonglü 5 were used as mapping populations.Fine mapping for vrnl11 was completed by using BSA sequencing technology and map-based cloning strategy.Phenotype identification results showed that,compared with wild-type WK3,the leaf width and leaf area of vrnl11 decreased by 25.7% and 21.7%,respectively.Genetic analysis showed the narrow leaf phenotype of vrnl11 was controlled by a single recessive nuclear gene.BSA sequencing analysis located the mutation site within a 4.7 Mb interval from 15.0 Mb to the chromosomal end on chromosome 11.vrnl11 was finally located in the 186.5 kb interval between the markers nl-61 and nl-46 by using these newly developed polymorphic molecular markers.The mapping interval contained 9 predicted genes.These results provide a theoretical basis for cloning vrnl11 and understanding the molecular regulation mechanism of leaf development in mungbean.

In order to investigate the structural characteristics of Brassica juncea AHK family proteins and their regulatory functions related to seed weight traits,it employed bioinformatics methods to perform whole-genome identification,physicochemical properties analysis,phylogenetic tree construction,protein structure analysis,promoter cis-elements analysis,and differential tissue expression profiling of the BjAHK family members.The main findings were as follows:a total of 19 BjAHK protein sequences were identified from the B.juncea genome,and phylogenetic analysis classified the BjAHK family members into four branches:G1 (BjAHK2—BjAHK4),G2 (BjAHK5),G3 (BjAHK1),and G4 (BjCKI1).Analysis of functional domains revealed the presence of six conserved motifs (Motif1—Motif6) in BjAHK family members,corresponding to the HisKA,HATPase_c,and REC conserved domains.The N-terminal regions of BjAHK1—BjAHK4 and BjCKI1 proteins exhibited relatively conserved transmembrane domains,which might be associated with their transmembrane function.Differential tissue expression profiling revealed that the genes encoding cytokinin receptor proteins (BjAHK2—BjAHK4) were constitutively expressed genes,with the highest expression levels observed in root tissues.Through integration with existing transcriptomic data on seed weight traits in B.juncea,four candidate genes potentially involved in the regulation of seed weight traits were identified:BjA07.AHK3,BjB03.AHK3,BjA10.AHK5,and BjB05.AHK5.BjA07.AHK3 and BjB03.AHK3 might positively regulate seed development,while BjA10.AHK5 and BjB05.AHK5 might have the opposite effect.Based on the integration of gene family analysis and transcriptome result,it was inferred that BjAHK3 and BjAHK5 genes play an important role in the regulation of seed development in B.juncea.

12-oxo-phytodienoic(OPR) acid reductase is a flavin mononucleotide-dependent oxidoreductase,a key enzyme for the synthesis of jasmonic acid,which is important for plant growth, development and defence regulation.In order to study the disease resistance effect of OPR gene in maize, bioinformatics methods were used to identify OPR family members in 31 different maize inbred lines and the expression pattern after infestation by S. turcica was analyzed.The results showed that eight OPR genes were identified in maize line B73 and they were unevenly distributed on seven chromosomes.In addition,these expressed proteins of OPR genes were rich in acidic amino acids.Further analysis revealed that the maize OPR family contained only one structural domain,Oxidored_FMN,and all OPR members contained 10 protein conserved motifs identified.Phylogenetic relationship analysis of OPR family members from maize,wheat,rice and Arabidopsis using MEGA software revealed significant differences in the evolutionary relationships of OPR gene families in these plants.The OPR proteins of maize and those of rice were the closest in evolution.Homology analysis using OrthoFinder software revealed that the maize OPR gene family was highly conserved,with all OPR genes being core genes,but with slightly different functions.According to the previous RNA-seq data of Hebei key Laboratory of Plant Physiology and Molecular Pathology,we analyzed the expression pattern of OPR genes of maize B73 in response to S.turcica infection and verified the gene expression pattern of OPR genes through qRT-PCR.It was found that there were three different expression patterns in the process of S.turcica infection.This study systematically identified the maize pan-genomic OPR family genes and their expression patterns in response to S.turcica infection.

Salt-alkali stress has become one of the important factors restricting agricultural production in my country.Exploring the molecular mechanism of salt tolerance of crops has important theoretical and practical value for crop breeding.The purpose of this study is to clone the ZmMPI gene in corn and transform corn plants.First,qRT-PCR was used to analyze the ZmMPI expression changes in plants treated with saline-alkali solutions.Then DNAMAN software was used to perform multiple comparison analysis of the ZmMPI protein sequence.MEGA 7.0 software was used to construct a phylogenetic tree,and a series of software were used to analyze the ZmMPI protein sequence.ZmMPI performed bioinformatics analysis.Finally,molecular cloning technology was used to successfully clone the coding sequence of the ZmMPI gene,construct a plant overexpression vector,and use Agrobacterium-mediated genetic transformation method to transform the corn inbred line B104.The overexpression transgenic plants were transformed at the genome level,transcription level and protein level.Identify and analyze changes in expression levels.The results showed that the expression level of the ZmMPI gene showed an overall trend of first increasing and then decreasing after being subjected to salt-alkali stress;the ZmMPI protein sequence comparison result showed a similarity rate of 64.15%,and the phylogenetic tree showed that ZmMPI had the highest homology with Zea mays subsp.parviglumis ABA34115.1.The protein contained a protein domain Potato_inhibit,which had an α-helix,a random coil and a β-turn.It was relatively hydrophobic and had 10 predicted Potential phosphorylation sites;the identification results of the 49 transformation events obtained showed that the ZmMPI gene in 13 over-expressed transgenic lines could be expressed normally at the genome level,and the ZmMPI gene in 10 over-expressed transgenic lines could be transcribed and translated normally.Finally,10 overexpression transgenic lines capable of normal transcription and translation were obtained,laying the foundation for further exploring the molecular mechanism of ZmMPI gene in response to salt-alkali stress.

To explore the function and possible molecular mechanisms of amino acid transporters (AATs),firstly,a new hidden markov model was established to identify the members of the rice AAT family,and a phylogenetic tree was constructed to analyze the evolutionary relationship of AAT members between rice and Arabidopsis;secondly,the promoter,protein structure,and functional domain of rice polyamine transporter gene 1 (OsPUT1) were analyzed using bioinformatics methods;thirdly,the subcellular localization of OsPUT1 was determined by constructing the OsPUT1-GFP vector and expressing it in rice protoplasts;fourthly,the expression patterns of OsPUT1 gene in different tissues and under different stress conditions were detected using qRT-PCR;finally,the function of OsPUT1 gene was preliminarily studied using the OE,Nip and RNAi lines.The results showed that there were 96 OsAATs divided into 13 subfamilies in rice;the PUT subfamily contained 6 OsPUTs,among which OsPUT1 and AtPUT3 had the closest genetic distance and were distributed in the same branch.The promoter of OsPUT1 gene contained cis-acting elements related to growth and development,light regulation,plant hormones,and stress response;OsPUT1 protein contained the polyamine transporter domain PotE,and subcellular localization experiment indicated that it was located on the cell membrane;the expression of OsPUT1 gene was relatively high in leaves,while was low in flowers;gene expression was inhibited by JA,mannitol,and ABA,while decreased and then returned to normal under low temperature stress,yet increased and then decreased under SA,Spm and paraquat treatment,however,under sodium chloride treatment,the gene expression level first increased,reached its peak at 1 h,then decreased,reached its lowest at 12 h,and returned to normal levels at 24 h;OE lines significantly reduced resistance to paraquat,while RNAi lines significantly increased resistance to paraquat.Overall,the OsPUT1 protein might have the function of transporting PA and participating in stress response.

JAZ protein plays a key role in plant growth and development and stress signaling pathways.In order to explore the regulatory mechanism of JAZ protein in wheat late spring cold,TaJAZ6 gene was cloned from young spike of wheat,and its molecular characteristics,expression characteristics and subcellular localization were analyzed.The results showed that the full-length CDS sequence of the gene was 549 bp,encoding 178 amino acids.The predicted molecular weight of the encoded protein was 18.376 ku,the theoretical isoelectric point was 9.37,and the instability coefficient was 62.44,so it was an unstable protein.The protein encoded by this gene had a TIFY domain and a CCT_2 domain.Phylogenetic tree analysis showed that the protein had the closest relationship with TIFY 11b proteins of Triticum dicoccoides and Triticum urartu.In addition to the basic response elements such as CAAT-box and TATA-box,the promoter region of TaJAZ6 gene also contained hormone response elements,light response elements,low temperature response elements,defense and stress response elements.Real-time Quantitative PCR analysis showed that TaJAZ6 gene was expressed in roots,stems,leaves and young panicles,with the highest expression in roots.The expression of TaJAZ6 gene was also induced by low temperature and methyl jasmonate (MeJA).Under low temperature stress,the expression trends of TaJAZ6 in roots,stems and leaves of Aikang 58 (tolerant to late spring cold ) and Zhengmai 366 ( sensitive to late spring cold ) were the same trend,which significantly increased.After spraying 300,350 μmol/L MeJA,the expression of TaJAZ6 in plants treated with low temperature decreased significantly in both wheat varieties.The expression level of TaJAZ6 in the young panicles after low temperature stress showed an opposite trend.The expression level of TaJAZ6 in the young panicles of Aikang 58 decreased significantly,and increased significantly in the young panicles of Zhengmai 366.It was speculated that the gene might negatively regulate the defense response of wheat to late spring cold stress.By spraying MeJA,the relative expression of TaJAZ6 gene in young spikes of two wheat varieties under low temperature stress was significantly reduced,and the grain number of wheat was increased.Subcellular localization assay showed that TaJAZ6 protein was localized in the nucleus.The results above indicate that TaJAZ6 may play an important role in the response of wheat to late spring cold stress.

To explore the physiological response mechanism of maize to different drip irrigation quotas,a 2-year pond planting experiment was conducted under controlled conditions.Using two maize varieties with differences in drought resistance as materials,six treatments were set up:CK1 (Drought resistant variety,500 mm),T1 (Drought resistant variety,350 mm),T2 (Drought resistant variety,200 mm),CK2 (Drought sensitive variety,500 mm),T3 (Drought sensitive variety,350 mm),and T4 (Drought sensitive variety,200 mm) to analyse the photosynthetic,chlorophyll fluorescence,photosynthetic response characteristics,and grain filling characteristics of maize leaves changed in hormone content,starch synthase activity,and yield in grains.The results showed that four photosynthetic parameters,including net photosynthetic rate (Pn),and four chlorophyll fluorescence parameters,including maximum photochemical quantum yield of photosystem Ⅱ (Fv/Fm),decreased with the decrease of drip irrigation quota,while stomatal limitation percentage (Ls) and non-photochemical quenching coefficient (NPQ) increased.The apparent quantum efficiency (AQE) and other 10 photosynthetic response related parameters of maize leaves all decreased with the decrease of drip irrigation quota.The range of differences between light compensation point (LCP) and light saturation point (LSP),CO₂ compensation point (CCP) and CO₂ saturation point (CSP) in CK1,T1,and CK2 treatments was larger than that in other treatments.The grain filling rate reached its peak at 25 d after anthesis,and the T2,and T4 treatments significantly decreased compared to the CK1 and CK2 treatments.The content of cytokinin (CTK) and auxin (IAA) in grains decreased with the decrease of drip irrigation quota,and the content of abscisic acid (ABA) increased.The activities of acid sucrose invertase,sucrose synthase,starch synthase and adenosine diphosphate glucose pyrophosphorylase in grains of T2 and T4 treatments were significantly lower than those of CK1 and CK2 treatments.The maize yield significantly decreased with the decrease of drip irrigation quota,and the T1 treatment only decreased by 3.45% to 4.51% compared to the CK1 treatment.There was no significant difference between T1 treatment and CK1 and CK2 treatment in the above indicators.The leaves of maize treated with T1 treatment still maintained the photosynthetic performance and photosystem Ⅱ structure,enhanced the adaptability of leaves to light and CO₂,increased the content of hormones related to maize grains and the activity of enzymes related to starch synthesis,effectively regulated the growth and development of maize grains and the filling process,and maize yield and its components performed better.

In order to reveal the influence mechanism of sowing date adjustment on crop yield formation, a field experiment was conducted to adjust the sowing date of the same variety of summer maize from 2019 to 2021 at Hebei Gucheng Agricultural Meteorology National Observation and Research Station in Northern north China, which was set up in four sowing date, as 10 d early sowing, 10 d late sowing, 20 d late sowing and control. By monitoring the growth period changes, plant dry matter accumulation, leaf photosynthetic characteristics and grain filling characteristics, and sampling at maturity to determine the yield components, the results showed that:the sowing date of summer maize was appropriate to be advanced,the seedling stage,ear stage and the whole growth period were extended,especially,the duration of effective filling was extended with the advance of sowing date.The effective grain filling duration days were extended by 4.7 days for every 10 days ahead of the sowing date.The average filling rate of winter wheat was 4.04% higher on the sowing date was advanced by 10 days, compared with the sowing date was deferred by 10 or 20 days and the control.Grain filling accumulation increased,and 100-grain weight increased by 5.459 g.The main yield components of summer maize,such as grain number per ear,grain weight per ear and 100-grain weight showed significant differences during sowing,and increased with the sowing date.The theoretical yield increased by 1 395.4 kg/ha with 10 days ahead of the sowing date during the experimental period from June 8 to July 8.The average net photosynthetic rate(Pn)of leaves in the critical development period of summer maize increased by 0.764 μmol/(m²·s)with every 10 days ahead of sowing date,and the average Pn of sowing date 10 days earlier was increased by 7.31% compared with the control and late sowing 10 and 20 days.With the increase of photosynthetic rate,dry matter production,accumulation and transfer to grain were increased,and the grain weight per ear and 100-grain weight increased by an average of 24.01% and 18.00%,respectively,compared to the control and late sowing 10 and 20 days.Advancing sowing date resulted in low plant height,strong stems,lodging resistance,large individual green leaf area,high leaf area index(LAI),and high leaf photosynthesis ability,above-ground dry matter allocation rate at maturity stage every 10 days in advance of sowing date:grain weight improved by 2.26%,the source—pool distribution of dry matter between plant vegetative organ and ear changed,ear weight per ear,100-grain weight and grain yield increased.The research showed that the double cropping area of winter wheat—summer maize in north China could effectively improve the yield per unit area by making full use of the heat resources increased by climate warming,reasonably allocating stubble,sowing summer maize early at the appropriate time,and extending the development period and grain filling time.

In order to clarify the main agronomic traits and their relationships with grain yield of different colored-grain wheat varieties(lines),and select colored-grain wheat varieties suitable for cultivation in Eastern Shandong.The experiment was carried out during 2020-2022 winter wheat growing seasons.Four purple-grain wheat varieties (lines)Qingyan No.1 purple wheat (QYZ-1),QYZ-2,Shannong No.1 purple wheat (SNZM1),and Nongda 3753 (ND3753),two blue-grain wheat lines 20064 and 20072,and one common white grain wheat variety Jimai 22 (JM22,control variety) were selected as test materials,to systematically study the differences in flag leaf SPAD,leaf area index,dry matter accumulation and remobilization,grain yield and its component factors,stability of agronomic traits and yield sustainability of different colored-grain wheat varieties (lines).The results showed that grain yield,thousand grains weight,leaf area index at anthesis,SPAD value after anthesis,dry matter accumulation at anthesis,dry matter accumulation at maturity,dry matter accumulation at post-anthesis and harvest index of each colored-grain wheat varieties (lines) were lower than those of common white grain wheat (Jimai 22).Compared among the various colored-grain wheat varieties (lines),the purple-grain wheat QYZ-1 grain yield was significantly higher than that of other colored-grain wheat varieties (lines),and the grains per unit area number was not significantly different from that of QYZ-2,but its thousand grains weight was significantly higher than that of QYZ-2.The upper three leaves area index of the purple-grain wheat QYZ-1 was significantly higher than that of the blue wheat lines and ND3753 at the anthesis stage.The flag leaf SPAD,the dry matter accumulation at maturity and post-anthesis,the dry matter remobilization before anthesis,and the harvest index were higher than those of other colored-grain wheat varieties (lines);compared with other varieties (lines),the coefficient of variation (CV) of agronomic traits of JM22 and QYZ-1 were lower.Compared with colored-grain wheat,the mean yield and yield sustainability index (SYI) of QYZ-1 were higher.In addition,the correlation analysis showed that the grain yield was positive significantly correlated with the dry matter accumulation at anthesis,the dry matter accumulation at maturity and post-anthesis,the dry matter remobilization before anthesis,the harvest index,flag leaf SPAD value at 28 days after anthesis,whole green leaves area index at anthesis and thousand grains weight,respectively.Based on the results of the two years' study,QYZ-1 obtained the appropriate leaf area index,maintained the higher flag leaf SPAD after anthesis,and the longer duration of anthesis to maturity stage,delayed the leaf senescence during the late grain filling stage,and synergistically increased the post-anthesis dry matter accumulation and the dry matter remobilization before anthesis,dry matter accumulation at maturity and harvest index,and grain number per unit area and thousand grains weight,and then achieved higher grain yield.In summary,Qingyan No.1 purple wheat,as a colored-grain wheat variety,has a stable yield and good sustainability,making it suitable for planting in Eastern Shandong region.

To clarify the effects of drought stress and cultivars on the net photosynthetic rate(Pn),the activities of photosynthetic enzymes in flag leaf in the afternoon(FLA)during grain filling stage and grain yield of winter wheat,a pond culture experiment with four water levels and two winter wheat cultivars was conducted under the condition of rainproof pond cultivation in 2019—2021.The four water treatments included severe drought(W1),moderate drought(W2),mild drought(W3),and suitable water supply(W4).The Pn,and the activities of Rubisco,RCA,PEPC,ATPase and PPDK in flag leaf from 14:00 to 16:00 during the early(EGFS)and medium(MGFS)grain filling stage,and the grain yield at maturity of strong drought resistance cultivar Jinmai 47(JM47)and weak drought resistance cultivar Yanzhan 4110(YZ4110)were investigated.Drought stress decreased the FLA Pn and activities of most photosynthetic enzymes,and the grain yield of wheat.The decrease of these indexes increased with drought stress degree,but the effects were different among varieties and years.Compared with W4,the FLA Pn of JM47 under W1,W2 and W3 decreased by 33.6%—40.6%,12.0%—30.5% and 5.0%—13.5%,as well as YZ4110 decreased by 44.0%—52.0%,22.5%—38.1% and 11.5%—20.5%,respectively.Compared with W4,the FLA Rubisco activity decreased during EGFS but increased during MGFS for JM47,while it decreased by 13.3%—25.6%,7.1%—14.0% and 11.2%—11.6% for YZ4110,respectively,under W1,W2 and W3.Compared with W4,the FLA RCA activity significantly decreased under most drought treatments during EGFS,while increased under W2 and W3 for JM47 and decreased under W1 and W2 for YZ4110 during MGFS.Compared with W4,the FLA ATPase activity of JM47decreased under W1 but increased under W3,while that of YZ4110 decreased under W1,W2 and W3 by 19.3%—48.7%,7.2%—24.2% and 0.1%—8.9%,respectively.The FLA PEPC activity under different treatments varied with growing seasons and varieties.Compared with W4,the FLA PPDK activities of JM47 and YZ4110 under W1 were decreased by 12.4%—18.8% and 16.7%—18.2%.Compared with YZ4110,in most conditions,the FLA Pn and photosynthetic enzyme activities of JM47 had no significant difference under suitable water supply(W4),but increased under drought treatments(W1,W2 and W3).The results of correlation analysis showed that yield,FLA Pn were significantly positively correlated to FLA ATPase activity during EGFS and MGFS,as well as FLA PEPC activity during EGFS.Thus,increasing the FLA ATPase and PEPC activities during the grain filling period is conducive to the increase of FLA Pn and grain yield of wheat.

To investigate the molecular response to drought stress of silks,two treatments with soil moisture content of 80% field holding capacity (CK) and 60% field holding capacity (DS) were set up using Annong 591 and Xianyu 335,and the maize silks were sampled during the rapid increase in silk elongation,and transcriptome sequencing was performed.By comparing the gene expression differences between the drought and control groups of the two varieties,the key pathways and related candidate genes of maize silks in response to drought were clarified.The results showed that the phenylpropane biosynthesis pathway (ko00940) and starch and sucrose metabolism pathway (ko00500) were the key pathways in maize silk response to drought.In the phenylpropane biosynthetic pathway 4CL,CCR,CAD and POD genes were lowly expressed in drought-prone maize silks.Correlation analysis showed that down-regulation of 4CL,CCR,CAD and POD genes was associated with reduced lignin content.Converting enzyme and sucrose synthase genes in the starch and sucrose metabolic pathways were lowly expressed in drought maize silks.Correlation analysis indicated that down-regulation of convertase and sucrose synthase genes was associated with sucrose metabolism.In summary,this study identified key genes and metabolites involved in sucrose and lignin metabolism.The lignin synthesis are associated with drought tolerance in maize silks,and 4CL, CCR,CAD,and POD are the candidate genes for drought tolerance in maize silks.In addition,the starch and sucrose metabolic pathways are associated with silk elongation and drought tolerance of silks,and invertase and sucrose synthase are the main candidate key enzymes.

In order to explore the suitable phosphate fertilizer application methods in the Huang-Huai area,this research conducted field experiments to investigate the dry matter accumulation,root morphological indexes,available phosphorus content and phosphatase activity in different soil layers,maize yield and its component factors of four phosphate fertilizer application methods(conventional broadcast (P1),layered application (P2),strip application (P3) and hole application (P4)).The results showed that the ear length and kernel row number of P2 and P4 were significantly increased by 22.57%,16.81% and 15.19%,7.60% compared with P1.The grain number per spike of P2,P3 and P4 were significantly increased by 25.26%,13.86% and 17.00% compared with P1,respectively.Additionally,a decrease of spikes per area was found in P2 (15.30%) compared with P1.The yields of P2 and P4 were significantly increased by 15.20% and 10.79% compared with P1.Among root traits,the root length,root surface area,root volume and root tip number were significantly affected by the phosphorus application method.The root length under P2 and P4 treatments was significantly increased by 30.41% and 33.75% compared with P1.The root surface area of P2 treatment was significantly increased by 23.77% compared with P1.The root surface area of P4 treatment was significantly increased by 29.60% and 21.70% compared with P1 and P3 treatments,respectively.The root volume and root tip number of P2 and P4 treatments were significantly higher than those of P1 and P3.The analysis of soil available phosphorus content showed that the available phosphorus contents of P2 and P4 were significantly higher than that of P1 in the soil layer of 10—20 cm at three-leaf stage of maize.The available phosphorus contents of P2,P3 and P4 in the 0—20 cm soil layer were lower than that of P1,and the available phosphorus content of P4 in the soil layer of 20—30 cm was significantly lower than that of P1 at maturity stage.Correlation analysis showed that there was a significant positive correlation between acid phosphatase activity and soil available phosphorus content in the 10—20 cm soil layer at three-leaf stage of maize.The acid phosphatase activity and soil available phosphorus content in P2 and P4 treatments were higher in this soil layer,which was conducive to the conversion of soil phosphorus into maize absorbable state.In conclusion,compared with the traditional application method,layered phosphorus application and cave phosphorus application can improve the soil phosphorus availability in the early stage of maize,promote the growth and development of roots,and then increase the summer maize yield,so they are more suitable application method for corn phosphorus fertilizer in Huang-Huai area.

In order to study the biological function of NbEHD1 in Nicotiana benthamiana,its gene structure,conserved domains,phosphorylation sites,subcellular localization and evolutionary relationship were predicted by bioinformatics methods.The encoding sequence of NbEHD1 was 1 638 bp,and its genomic sequence contained 16 exons and 15 introns.NbEHD1 protein was predicted located in the cytoplasm,which had 42 phosphorylation sites and had no signal peptide and transmembrane region.NbEHD1 belonged to the P-loop_NTPase superfamily and had a conserved domain specific to the EHD family.Phylogenetic relationship showed that NbEHD1 was closely related to EHD sequences of Solanum tuberosum and Solanum lycopersicum.Blast analysis against N.benthamiana sub-data base in SNG was performed to obtain the predicted full length of NbEHD1,then its full nucleic acid sequence was amplified by gene-specific primers.After obtaining NbEHD1 CRISPR/Cas9 gene editing construct,the vector was successfully transformed into the leaves of N.benthamiana by Agrobacterium-mediated genetic transformation.Eventually 16 T₀-generation plantlets were successfully identified,which provided materials for further research to determine the biological function of NbEHD1.

To investigate the function of GhFUL1 gene in branching development of cotton,the GhFUL1 gene in the MADS-box family and its promoter were cloned from cotton,and the function and promoter activity of the gene were analyzed.The results indicated that the open reading frame (ORF) of GhFUL1 gene was 726 bp and encoded 241 amino acids.Phylogenetic tree analysis showed that the amino acid sequence of FUL subclade was conserved,and GhFUL1 owned the highest similarity to Cacao TcAGL8-1 except for the homologous proteins in cotton.Subcellular localization result showed that GhFUL1 was localized in nucleus and membrane.The expression level of different cotton tissues was examined,and the result showed that GhFUL1 was highly expressed in shoot apical meristem.Overexpressing GhFUL1 in Arabidopsis thaliana increased the number of lateral branches by decreasing the expression of cytokinin oxidation/dehydrogenase genes AtCTK1 and AtCTK6.Compared with the wild type,transgenic plants did not result in significant changes of the flowering time and floral organ development.The present results suggested that GhFUL1 might affect the number of branches by regulating the cytokinin synthesis pathway.The promoter of GhFUL1 was predicted to include the TATA-box and CAAT-box elements,and cis-acting elements related to light response,auxin response and stress response by PlantCARE website.Then the promoter of GhFUL1 was cloned and constructed into expression vector pBI121 to detect promoter activity.The GUS staining of transgenic Arabidopsis lines showed that GhFUL1 promoter was specifically expressed in shoot apical meristem at seedling stage and flower organ at maturity.This study revealed that the gene and promoter of GhFUL1 played an important role in the Arabidopsis branching development process after transformation.

In order to promote the breeding of high quality grain shape Japonica rice varieties,a natural population consisting of 295 Japonica rice varieties collected from around the world was used as test material for phenotypic analysis of five grain shape-related traits,including grain length,grain width,grain thickness,aspect ratio and thousand grains weight,between 2018 and 2019,combined with 788,396 SNPs obtained by resequencing,and genome-wide association analysis using the MLM model of TASSEL 5.0.Haplotype analysis was performed on all the genes in the QTL intervals that were jointly detected during the two years and controlled multiple shape-related traits, and candidate genes for high quality rice grains were mined by combining the results of previous studies and functional annotation of the genes. The results showed that there was extensive phenotypic variation for the five grain shape-related traits,all of which were approximately normally distributed,and most of the grain shape-related traits were significantly or highly significantly correlated with each other. And a total of 221 QTLs significantly associated with rice grain shape-related traits were detected under the threshold condition of P≤5.46×10^-6,which were distributed on all 12 chromosomes of rice,and phenotypic contribution was 8.62%-20.73%, of which, seven QTLs were jointly detected in two years, and combined with the haplotype analysis and functional annotation of the genes, it was hypothesized that LOC_Os12g 44290 was a new candidate gene for rice grain shape. In summary,this study used genome-wide association analysis for QTL localization and candidate gene analysis for grain type-related traits in 295 Japonica rice varieties,providing a theoretical basis for breeding Japonica rice varieties with high quality grain types.

The plant NADPH oxidase Rbohs(Respiratory burst oxidase homologs) is the main source of reactive oxygen species (ROS),which participate in various physiological processes such as plant growth,development,stress resistance and plant-microorganism interaction.In order to explore the function and mechanism of Rbohs in symbiotic nitrogen fixation,GmRbohL,a member of soybean Rbohs gene family,was cloned in this study.The gene expression pattern,protein subcellular localization and gene function were studied by molecular biology,cell biology and genetics,respectively.The results revealed that: GmRbohL gene was induced by rhizobia and expressed specifically in soybean roots and nodules.Subcellular localization analysis indicated that the gene-encoded protein GmRbohL was a membrane protein.The plant gene silencing (RNAi) vector of GmRbohL was constructed,and the transgenic hairy roots were obtained by the transformation of soybean hairy root mediated by Agrobacterium rhizogenes K599.Gene silencing of GmRbohL resulted in a significant reduction in the number of nodules of transgenic hairy roots,and the production of ROS was also inhibited. Gene silencing of GmRbohL reduced the infection of rhizobia at the stage of root nodule organogenesis,and the expression level of nodulation marker genes also decreased with the decrease of GmRbohL expression.The root nodule tissue sections showed that gene silencing of GmRbohL significantly reduced the number of symbionts in the infected area of root nodules,and the nitrogenase activity of root nodules also decreased accordingly.The above data indicated that gene silencing of GmRbohL significantly inhibits the symbiotic nodulation process of soybean by reducing the production level of ROS.It is speculated that GmRbohL may play an important positive regulatory role in organogenesis of soybean nodules and regulation of nitrogen fixation function.

Sucrose is the main form of water-soluble carbohydrates transported in wheat stems,and plays a key role in the regulation of wheat growth and grain filling,which is a typical quantitative trait with a complex genetic basis.The SNP loci and candidate genes significantly associated with sucrose accumulation and translocation in wheat stems were explored at the genome-wide level to provide a theoretical basis for gene cloning and molecular marker-assisted selection.Seven traits of 117 domestic wheat varieties were measured, such as the sucrose content of flowering, filling, and maturity, the translocation rate and contribution rate to grain filling of sucrose at the pre-anthesis and post-anthesis, under different environmental conditions. Based on 35K SNP microarray genotyping, a mixed linear model MLM (Q + K) was used for genome-wide association study (GWAS) and candidate gene prediction.The overall phenotypic variation ranged from 9.53% to 45.70% with rich phenotypic diversity;the heritability of seven traits ranged from 0.32 to 0.49;the genotyping results showed that the polymorphism information content (PIC) of SNP markers ranged from 0.10 to 0.38;the population structure analysis divided the tested materials into four subgroups.The MLM(Q+K)model detected 146 significantly associated loci for traits related to sucrose accumulation and transport,among them,three SNP loci were detected repeatedly in two environments,and 13 candidate genes related to sucrose accumulation and transport related traits were screened based on the 200 kb physical location interval upstream and downstream of the 3 SNP loci that were stably and significantly associated.Among them, TraesCS1B02G324100 (Nucleotide-diphospho-suger transferase gene),TraesCS1B02G324200(Cellulose synthase-like protein gene),and TraesCS1B02G324300 (β-1,3-glucanase gene) were associated with sugar metabolism.

Spike length is an important agronomic trait in wheat and is closely related to yield components.Studying wheat spike length genes and screening molecular markers linked to spike length genes can provide molecular support for wheat molecular marker-assisted breeding.The F₂ population constructed with Keda 116 and Keda 101 as parents was used as test material to construct a genetic map covering the wheat genome by SSR molecular markers and to locate QTLs for spike length by combining the complete composite interval mapping method.A total of 434 pairs of primers with polymorphism between parents were screened from 3 234 pairs of primers,and the detection rate of polymorphic primers was 13.42%.A total of 28 molecular markers were screened for possible linkage to spike length by BSA mixed pool analysis,16 of these markers were verified to be tightly linked to the target gene by a population of 262 plants.The genetic map of wheat chromosome group was constructed by QTL-IciMapping software,and the average genetic distance between markers was 38.66 cM.A total of seven QTLs loci associated with spike length were detected,which were located on chromosomes 3B,4A,4B and 6B.The additive effect values of all the seven QTLs were positive and their contributions to the genetic variation of phenotypic traits ranged from 4.01% to 23.16%.Two major QTLs were mapped on chromosome 4B,explaining 17.59%-23.16% of the phenotypic variance.Among them,Qsl4B-2 was the most closely linked QTL locus with only 3.5 cM away from the nearest molecular marker,and the analysis found that it might be a new major QTL locus.Therefore,genes associated with spike length might exist on chromosome 4B.Within the marker interval of chromosome 4B from yzu397456 to yzu404917 and yzu409422 to yzu405167,there might be seven candidate genes regulating wheat spike length,which were consistently highly expressed in the spike.

In order to study the correlation between stem traits and the formation of elasticity and density tolerance of maize,and further reveal the mechanism of lodging resistance,6 maize varieties with different lodging resistance were selected as materials,and 3 planting densities of 6.0×10⁴,7.5×10⁴ and 9.0×10⁴ plants/ha were set.The stem pull lodging angle in the field was used as the evaluation index of elasticity,and the morphological characteristics between plants and basal nodes.The internode anatomical structure,material accumulation and mechanical characteristics were analyzed.The results showed that plant height,ear height,basal internode length,thickness,epidermal thickness,crusty tissue thickness,total vascular bundle,sheath area of small vascular bundle,fresh weight per unit length,dry weight per unit length and content of each component,puncture and folding strength had significant effects on the stem pull lodging angle,among which internode diameter(r=0.521^**)and dry weight per unit length(r=0.562^**)had the greatest effects.The greater the planting density,the smaller the internode diameter,the thickness of the hard skin tissue,the total number of vascular bundles,the fresh weight per unit length,the dry weight,the content of each component,the lodging resistance,the worse the elasticity of the stem.There were significant differences in stem traits among different varieties.The elasticity related traits of Lishou 1,Chuangyu 107,Jingnongke 728 and MC278 were better than those of other varieties,and the variation amplitude was smaller with the increase of density,so the elasticity and density tolerance were stronger.Traits such as internode diameter and dry weight per unit length had significant effects on stem pull lodging angle, that was stalk elasticity,and the variation amplitude of these traits after densification determined the density tolerance of stalk.

In order to further explore the primary QTL loci for grain-related traits in wheat and explore the genetic relationships among grain traits,124 DH populations constructed from wheat varieties AN859 and WN988 with large differences in grain traits were utilized as research materials,The phenotypic values of grain length,grain width,and thousand grains weight were measured in seven environments over two years,respectively,to carry out the multiple regression analysis of grain traits,and QTL detection of grain-related traits was performed based on the 55K microarray data of the DH populations.The results showed that grain width contributed most to thousand grains weight in the multiple regression analysis.QTL localization for grain traits by complete interval mapping,a total of 69 QTLs related to grain traits were detected on 19 chromosomes except chromosome 6D and 7B,including 24 QTLs for thousand grains weight,28 QTLs for grain length,and 17 QTLs for grain width,with phenotypic interpretations of individual QTLs ranging from 6.87% to 27.74%.Among them,grain length-associated Qgl.ahau-7A.1 on chromosome 7A was detected under seven environments and BLUP,with a phenotypic interpretation rate of 9.48%—22.26%,an additive effect of 0.11—0.21 mm,and a physical interval of 4.91 Mb(AX-110430243—AX-110442528),for the new primary effector QTL.Therefore,the Qgl.ahau-7A.1 locus can be used as a region of focus for subsequent fine localization and molecular marker-assisted breeding.

Conservation tillage and organic fertilizer application are one of the effective solutions to alleviate ecological vulnerability in dry farming areas of the Loess Plateau.Reasonable tillage and fertilization measures are of great significance for the realization of two crops a year in the arid area of central Shanxi.The dry farmland in the Loess Plateau was taken as the research object,and the split-plot experimental design was adopted,the main area was three tillage methods (deep tillage (DT),deep loosening (SS),no tillage (NT)),and the sub-area was four fertilization levels (No fertilizer control (CK),full chemical fertilizer (CF),50% chemical fertilizer+50% organic fertilizer (OF),full organic fertilizer (OM)).The changes of soil bulk density,available nutrients and annual yield of wheat and maize under different tillage and fertilization methods were explored.The results showed that in 0-20 cm the soil bulk density of SS and NT treatments were lower than that of DT treatment after organic fertilizer application.The soil bulk density of SS+OF treatment was 1.13 g/cm³,which was significantly lower than that of DT+OF treatment in wheat season.In the whole annual rotation system,the soil mass water content of NT+OM treatment was the highest,which was 7.88 percent point higher than that of other fertilization treatments,and the three-phase ratio of soil under SS+OM treatment was more ideal,and the deviation value of three phase was low.Under the three tillage methods,the content of available nitrogen and available phosphorus generally showed the trend of OM>OF>CF>CK,and the content of available potassium was significantly related to the fertilization method,which was the highest under the CF treatment.The application of organic manure in maize season significantly increased the fresh ear yield of maize compared with a single application of chemical fertilizer,but the difference between them was not significant.The annual yield of wheat and maize under SS+OM treatment was the highest,reaching 19 145 kg/ha.In summary,under the conditions of this experiment,the combination of SS,NT and organic fertilizer application could significantly improve the physical properties of the soil,and the content of alkali-hydrolyzable nitrogen and available phosphorus also had a certain degree of improvement,and SS+OM treatment was beneficial to the increase of the yield of wheat-maize continuous cropping field in the Loess Plateau.

To investigate the effects of Mn treatment on the growth,development and cadmium adsorption capacity of cotton under Cd stress.Different concentrations of Mn treatment(0,5,10,50 μmol/L)were applied to exploit changes of superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA),free proline(Pro),Cd content,dry matter quality,and main root length of roots of cotton seedlings of Xiang C178 and Xiang FZ001 under Cd stress.The stress of 10 μmol/L Cd had a significant effect on the growth of cotton root.The physical barrier and antioxidant defense mechanism were constructed to reduce the damage of Cd to root cells,resulting in a decrease in SOD activity and an increase in CAT activity,MDA content and Pro content.And, adding Mn was observed to effect the physiological response to cadmium of cotton seedling roots. Low concentration of Mn treatment resulted in the decrease of Cd content,the increase in SOD activity and Pro content,the decrease in CAT activity and MDA content in the root of cotton seedlings,which effectively prevented the inhibition of Cd stress on the root growth of cotton seedlings.High concentrations of Mn could increase Cd content and SOD activity in cotton seedling roots, which affected the root growth of cotton seedlings.When using cotton to repair Cd-contaminated farmland,it can be less than 10 μmol/L Mn ion treatment is used to improve the Cd tolerance of cotton seedlings and ensure the normal growth of cotton plants.

Partial substitution of organic fertilizer nitrogen for chemical fertilizer nitrogen is one of the ways to achieve sustainable crop development.This study explored the appropriate ratio of wheat organic nitrogen partial substitution for chemical nitrogen,as well as the characteristics of nitrogen accumulation,transportation,and utilization after substitution,in order to provide a basis for nitrogen fertilizer reduction and efficiency enhancement technology for winter wheat in Hebei Region.Field experiments of the following nine treatments were conducted in Ningjin,Hebei from 2021 to 2023:T1,non-nitrogen,and application of chemical P and K fertilizer alone;T2,high efficiency fertilization treatment,and application of chemical N,P and K fertilizer alone;T3—T7,substitution of 20%,40%,60%,80% and 100% the chemical N rate of T2 with organic fertilizer,respectively;T8,traditional fertilization,and application of chemical N,P and K fertilizer alone;T9,substitution of 100% chemical N rate of T2 with organic fertilizer,and spraying liquid nitrogen fertilizer at the erecting stage.The results of two years of experiments showed that the 100% substitution rate+liquid nitrogen treatment could achieve the highest wheat yield.Secondly,the yield of the 40% substitution rate treatment was equivalent to that of the high efficiency fertilization treatment,and it was much higher than that of the traditional fertilization treatment in the second year of the experiment.The 100% substitution rate+liquid nitrogen treatment increased the nitrogen content in stems and leaves by spraying available nitrogen during the erecting period,and the nitrogen accumulation in plants was equivalent to that of high efficiency fertilization and traditional fertilization treatments.Treatment with 40% and 80% substitution rates also achieved nitrogen accumulation equivalent to the highly efficient fertilization treatment.The 20%—100% substitution rate treatment (including liquid nitrogen treatment) could achieve a higher nitrogen transfer rate in stems and leaves,as well as the contribution rate of nitrogen transfer to grain.Among them,the 100% substitution rate+liquid nitrogen treatment had good fertilizer nitrogen absorption and utilization effect,achieving higher fertilizer nitrogen accumulation,nitrogen utilization rate,and nitrogen harvest index.Nitrogen fertilizer effect of the 100% substitution rate+liquid nitrogen treatment was equivalent to or slightly higher than that of highly efficient fertilization treatment.Secondly,the treatment with the 40% substitution rate had a similar or slightly lower nitrogen fertilizer effect than the highly efficient fertilization treatment.In summary,the 100% substitution rate+liquid nitrogen treatment showed better wheat yield,plant nitrogen accumulation,nitrogen transport rate,grain nitrogen accumulation,and nitrogen efficiency,followed by the 40% substitution rate treatment.

In order to explore the effect of salt stress on the physiological mechanism of cotton,saline-tolerant cotton Zhong J0710 was used as a control,two new cotton distantly hybrid germplasms HL2 and A₂H cultivated in cotton breeding research group were treated with salt solution for 15 d(concentration of 200 mmol/L)at seedling stage,and the relative plant height,relative root length and relative survival rate of the test materials were analyzed,and the salt damage index was calculated.The salt tolerance of the test materials was comprehensively evaluated by combining the measurement results of physiological indicators such as primary light energy conversion efficiency (Fv/Fm),antioxidant enzyme activity,proline content,and soluble sugar of seedlings at different treatment times (0,5,12,24,48 h).The results showed that after 15 d of salt stress,the damage degree of the growth of seedlings of the three test materials was as follows:Zhong J0710<HL2<A₂H,and salt stress significantly inhibited the relative plant height,relative root length and relative survival rate of A₂H,indicating that the salt concentration treatment had less effect on the growth and development of HL2 and Zhong J0710,but had a greater effect on A₂H.Under different treatment times,salt stress had different degrees of influence on the maximum light energy conversion efficiency of the three test materials,and the inhibition degrees from weak to strong compared with before stress were HL2<A₂H<Zhong J0710.Compared with the non-stress period, the peroxidase (POD) activity of the tested materials was increased by salt stress, and the POD activity of HL2 during the stress process was significantly higher than that of A2H and Zhong J0710 in the salt-tolerant control, and reached a very significant level at 12—48 h of stress,and the change law of superoxide dismutase (SOD) activity was not obvious,and the role in salt stress was low.The contents of proline in HL2 and the salt-tolerant control increased with the increase of salt stress time,and the soluble sugars and soluble proteins of germplasm HL2 were significantly higher than those in the salt-tolerant control and A₂H during the entire salt stress period.In summary,in salt adversity,salt-tolerant materials have a more developed root system;higher light energy conversion efficiency,higher reactive oxygen species clearance capacity in cells and higher accumulation of osmoregulatory substances are the physiological basis for strong salt tolerance.

Glutamate decarboxylase(GAD)is a cleavage enzyme that catalyzes the decarboxylation of glutamic acid,plays a key role in GABA synthesis pathway and has an important influence on plants' resistance to abiotic stress.Three watermelon GAD gene family members(Cla97C01G007270,Cla97C01G007290 and Cla97C04G079700)were screened by the association analysis of metabolomics and transcriptomes.On this basis,three watermelon GAD family genes were cloned,named ClGAD1,ClGAD2 and ClGAD3 respectively,and their bioinformatics and expression patterns were analyzed.The results showed that the CDS of ClGAD1,ClGAD2 and ClGAD3 were 1 524,1 497 and 1 497 bp,respectively,and the number of amino acids encoded by them was 507,498 and 498,respectively.All three proteins were hydrophilic proteins and were located in mitochondria.It was highly conserved,and the conserved domain was glutamic acid decarboxylase.Its promoter region contained a large number of CAAT-box and TATA-box,as well as corresponding elements related to adversity stress such as MYB and MYC.The secondary structures of the three GAD proteins were mainly α-helix and random coil.The tertiary structures were all homohexamer structures.Phylogenetic tree showed that members of GAD gene family were closest to GAD of Momordica charantia and Arabidopsis thaliana.qRT-PCR analysis indicated that the highest expression levels of ClGAD1 and ClGAD2 were showed in stems,while that of ClGAD3 was showed in flowers.The expression levels of ClGAD1,ClGAD2 and ClGAD3 were the highest under salt stress for 12,24 and 48 h,respectively.Under cold and drought stress,the expression patterns of the three GAD genes were similar,and all of them were expressed in large quantities in the early stage of stress,and the expression of ClGAD3 under abiotic stress was higher than that of the other two family members.The full-length cDNA of three GAD genes in watermelon was obtained,and their bioinformatics and expression patterns were analyzed,which provided an experimental basis for enriching the abiotic stress resistance gene resources of watermelon.

The domain of unknown function 668(DUF668)family is a family of plants whose function is unknown.To unveil the function and characteristics of maize DUF668(ZmDUF668)gene family,we identified ZmDUF668 gene family by bioinformatics method.The results showed that there were 19 ZmDUF668 genes in maize,distributing on 8 chromosomes,named ZmDUF668-1—ZmDUF668-19;the most of proteins encoded by ZmDUF668 were alkaline,and most of the family members were localized in the nucleus,cytoplasm and chloroplast.ZmDUF668 family proteins can be divided into two subfamilies according to the multispecies phylogenetic tree.Ten Motifs were identified from 19 members of ZmDUF668,all of which contained Motif 1 and Motif 5.Through the analysis of protein conserved domain,it was found that 14 of the 19 members contained not only DUF668 domain but also DUF3475 domain.Gene structure analysis showed that members of the same subfamilies had similar gene structure.Synteny analysis showed that there were 21 collinear relationships between 13 ZmDUF668 genes and 10 OsDUF668 genes.The analysis of cis-acting elements revealed that the promoters of ZmDUF668 genes widely contained cis-elements that light-response,plant hormones and abiotic stress.The prediction results of protein-protein interaction network(PPI)indicated that only ZmDUF668-10 of the ZmDUF668 family proteins had interaction with other proteins.Analysis of RNA-Seq revealed that the gene expression level of some ZmDUF668 gene family members changed significantly under cold,heat,salt stress and ultraviolet treatment.The response of ZmDUF668 family genes to cold and heat stress was verified by RT-qPCR.Bioinformatics was applied to the analysis of the ZmDUF668 gene family,unveiling the characteristics of the members of the ZmDUF668 gene family,and providing theoretical basis for the subsequent study of the molecular biological function of the ZmDUF668 gene family.

In order to explore the effects of amino acid water-soluble fertilizer on grape yield and quality,the full-fruit Zaohebao grape was used as the experimental material,three treatments were set up:conventional fertilization (CK);increased application of organic fertilizer (OPT);organic fertilizer + amino acid water soluble fertilizer treatment (OPT+AA),the effects of exogenous amino acids on grape yield,growth,quality and soil nutrient content were studied.The results showed that compared with CK,grape yield decreased by 14.17% under OPT treatment and 7.50% under OPT+AA treatment,but were not significant.Compared with CK,partial productivity of fertilizer treated with OPT and OPT+AA increased significantly.Partial productivity of N,P₂O₅ and K₂O treated with OPT+AA increased significantly by 73.44%,85.01% and 85.01%,respectively.OPT+AA significantly increased total sugar by 8.09%,soluble solids by 1.32 percentage,sugar-acid ratio by 38.80%,and the titratable acid content by 0.16 percentage.The nutrient content of shoots was significantly increased by 24.73% in OPT+AA,total phosphorus content and total potassium content were significantly increased by 24.73%.In addition,OPT and OPT+AA treatments did not inhibit plant growth,but could effectively reduce the leaching of available nutrients and increase the nutrient content in the root domain.In summary,reducing and increasing the application of organic fertilizer had no significant effect on grape yield,but could reduce the leaching of available nutrients and significantly improve the partial productivity of fertilizer.The application of amino acid water-soluble fertilizer could promote nutrient absorption in plants,improving grape yield and fruit quality.

In order to explore the response of ARP to low temperature stress in ornamental pepper,and provide a theoretical basis for subsequent research on gene function in ornamental pepper under low temperature stress.This study cloned homologous ARP gene from ornamental pepper using RT-PCR technology,named CfARP,and analyzed their expression in different tissues of ornamental peppers and under low temperature treatment;Using bioinformatics analysis to study gene protein coding,physicochemical properties,and genetic relationships,and using qRT-PCR technology to detect the expression level of CfARP in different tissues of ornamental pepper and under low temperature treatment.The results showed that the CDS sequence of the CfARP gene was 342 bp,with 100% homology with Zunla 1.It could encode 113 amino acids and contained a conserved Auxin-repressed domain;protein physicochemical analysis revealed that the molecular weight of CfARP protein was 12.156 ku,its isoelectric point was 10.22,and the average coefficient of hydrophilicity was -0.913.It was preliminarily predicted that CfARP was a hydrophilic protein;compared with other species' ARP amino acid sequences,it was found that CfARP was highly conserved in Solanaceae plants.The Real-time Fluorescence Quantitative PCR results showed that the expression level of CfARP gene was the highest in the stems of ornamental pepper,followed by roots,and relatively less in leaves and flowers;the expression level of CfARP gene continuously increased with the increase of low-temperature treatment duration.It was preliminarily speculated that CfARP had a certain role in the response of ornamental pepper to low temperature stress.

To express the protein StSPS1 encoding potato sucrose phosphate synthase(SPS)in E.coli and prepare polyclonal antibodies.The StSPS1 gene was cloned from the tubers of tetraploid potato variety Chuanyu 10,with a total coding region of 3 165 bp and a protein length of 1 055 aa.Subsequently,based on the constructed His tag fusion expression vector PET30a-StSPS1,the protein induction,denaturation,purification,renaturation,and rabbit immune tests of StSPS1 protein were carried out.The results showed that the molecular weight of StSPS1 protein was approximately 119.62 ku,and its expression was minimal in soluble supernatant,mainly in insoluble precipitates.The optimal induction conditions were induced with 0.5 or 1.0 mmol/L IPTG at 37 ℃ for 4 h.Due to StSPS1 being an inclusion body protein,it was subjected to inclusion body denaturation and purified using His tags to match the size of the target band.At the same time,a protein immunoblotting(WB)test was performed using His antibodies,and the target band was detected at 119.62 ku,indicating the successful purification of StSPS1 inclusion body protein.Finally,by injecting the dialyzed and refolded StSPS1 protein into the subcutaneous tissue of rabbits,two antibodies against StSPS1 were successfully immunized.After WB identification,it was found that both antibodies could hybridize target bands in the antigen and the total protein of leaves in Chuanyu 10.In summary,potato StSPS1 protein was induced and purified,and rabbit polyclonal antibodies against StSPS1 protein were successfully prepared.

Photoperiod is one of important environmental factors affecting plant bolting and flowering which are regulated by plant phytochrome proteins.The signal network mediated by PHYB has an important inhibitory effect on plant bolting and flowering.Previous studies revealed that there were large segment insertion/deletion differences between the PHYB gene promoter of the Chinese cabbage late-bolting line 06-247 and the easy-bolting line He102.In order to further investigate the impact of promoter mutation on PHYB and the key genes of its downstream pathways,this study was conducted.Based on bioinformatics methods,the redundancy characteristics of phytochrome genes in genome Chinese cabbage were firstly analyzed.It was found that the Chinese cabbage genome contained six phytochrome genes,of which PHYA had two copies,and PHYB,PHYC,PHYD,and PHYE all had only one copy.Then amino acid sequence alignment was used for screening of the specific sequence of PHYB.Antigenic determination clusters were designed based on the specific sequence and the antibodies against PHYB was prepared.The fluorescence quantitative RT-PCR technology and Western Blot technology were used to study the relative content of PHYB in 06-247 and He102.At the same time,the dynamic changes of key regulatory genes such as CCA1,FLC,CO and FT which in the downstream of PHYB pathway were also compared.The results showed that the promoter mutation caused significant differences both in level of mRNA in 06-247 and then significantly increased the protein level of PHYB.At the same time,the downstream regulatory genes such as CCA1,FLC,CO and FT were highly expressed in 06-247,which had an important impact on bolting resistance of 06-247.

In order to excavate the mycovirus resources in Ustilaginoidea virens and deeply analyze the relationship between the genome organization and function of a novel mycovirus,it took a U.virens strain Uv321 with abnormal phenotype,isolated from Hainan Province,as the research object,and identified the species of the novel mycovirus in the strain Uv321 on the basis of the previous meta-transcriptome data,a series of studies have been carried out around the novel mycovirus.The results showed that a novel mycovirus was identified in strain Uv321,named Ustilaginoidea viruses botourmiavirus 7 (UvBV7).The genome of UvBV7 is positive single-stranded RNA(+ssRNA),with a total length of 2 406 nt and a GC-content of 53.78%,containing an open reading frame(ORF)encoding RNA-dependent RNA polymerase(RdRP),which encodes 643 amino acids with a molecular weight of about 72.727 ku.The prediction of the protein secondary structure of the viral terminal showed that the 5' and 3' terminal bases of UvBV7 were complementary and paired,forming a hairpin structure.The BlastP alignment showed that UvBV7 had the highest similarity with the virus Erysiphe necator associated ourmia-like virus 72,which belonged to the genus Botoulivirus in the family Botourmiaviridae,but only 44.52%.The multiple alignment results based on the RdRP sequences of UvBV7 and other similar viruses showed that there were 8 conserved domains in the RdRP amino acid sequences of UvBV7 and the members of the family Botourmiaviridae.The GDD motif was found in the Ⅵ conserved domain,which is the typical highly conserved core motif of viral RdRP proteins.The phylogenetic tree constructed based on the amino acid sequence of the viral RdRP also indicated that UvBV7 clustered with the members belonging to the genus Botoulivirus.Therefore,UvBV7 is a novel mycovirus belonging to the genus Botoulivirus in the family Botourmiaviridae.The results of dual cultures of different strains of U.virens showed that UvBV7 could be transmitted horizontally between vegetative compatibility strains,but the mycelial tip-ribavirin,heat-ribavirin and protoplast regeneration-ribavirin treatments were unable to eliminate the mycovirus UvBV7 in strain Uv321.In conclusion,this study not only enriched the diversity of mycoviruses in U.virens,but also provided potential biocontrol agents with hypovirulence for the biocontrol of rice false smut.

To explore the influence of straw returning years to the field after deep turning back on soil bacterial community in continuous cropping corn fields in the West Liaohe Plain,and provide theoretical guidance for continuous straw returning to improve soil fertility.This study takes corn straw leaving the field as a control (CK),based on the high-throughput sequencing technology of bacterial 16S rDNA V3-V4 region,combined with bioinformatics,we analyzed the characteristics of organic matter,nitrogen nutrients content,soil bacterial community structure and diversity changes in 0-20 cm and 20-40 cm soil layers of corn fields with continuous straw returning for 2(SR2),5(SR5) and 10 a (SR10).The results showed that SR2 had no significant difference with CK in soil organic matter and nitrogen nutrients content,while SR5 and SR10 significantly increased soil organic matter and nitrogen nutrient content;under different straw returning years,soil bacterial diversity was different and each had its own unique OTU,among which SR10 OTU had the largest number;alpha diversity index showed that the relative abundance and diversity of soil bacterial community were not significantly different between SR2 and CK,SR5 and SR10 increased significantly,and SR10 and CK had the largest difference;a total of 51 bacterial groups were obtained at the phyla level,including Actinobacteriota,Proteobacteria,Acidobacteriota,Crenarchaeota and Chloroflexi,whose relative abundance was more than 5%.The relative abundance of Proteobacteria,Acidobacteriota and Chloroflexi increased with the increase of returning years.RDA clustering analysis showed that the soil bacterial community structure of 0-20 cm soil layer SR2 was similar to CK,SR5 was more similar to SR10,20-40 cm soil layer,different straw returning years,treatments were different from CK,SR2 and SR5 were similar,SR10 had significant differences in community structure compared to SR5 and SR2.The contents of soil {\mathrm{NO}}_3^{-}-N, {\mathrm{NH}}_4^{+}-N,OM,and TN all had effects on bacterial flora,and they were soil {\mathrm{NO}}_3^{-}-N> {\mathrm{NH}}_4^{+}-N>OM>TN.The contents of soil organic matter and nitrogen nutrient had no significant difference between continuous straw returning for 2 a and CK,while continuous straw returning for 5 and 10 a were significantly increased.In summary,the microbial diversity and richness of bacteria showed minimal changes in the soil layers of 0-20 cm and 20-40 cm after continuous straws returning for 2 a.After continuous straws returning for 5 and 10 a,both soil layers significantly increased;the bacterial community structure continuous straw returning for 2 a was similar to CK in the 0-20 cm soil layer,while the bacterial community structure continuous straw returning for 2,5,10 a were changed in the 20-40 cm soil layer.

To investigate the expression characteristics of the PDK4 gene in various tissues of Meiren yaks and to explore its correlation with the mechanism of fat deposition,tissue samples were collected from the longest and dorsal muscles,heart,liver,spleen,lung,kidney,fat and testis of Meiren yaks,and total RNA was extracted.The gene was amplified,analyzed and characterized by RT-PCR,bioinformatics software,and qPCR.The results showed that the CDS region of the PDK4 gene was 1 224 bp,407 amino acids,the relative molecular mass of protein was 46.159 14 ku,the theoretical isoelectric point was 6.60,and the total average hydrophilicity was-0.169,which was a hydrophilic protein;the phylogenetic tree results showed that Meiren yak was the closest related to wild yak and the most distant related to hyena;PDK4 protein had 34 phosphorylation sites,no signal peptide and transmembrane region,and mainly plays a biological function in mitochondria;the PDK4 protein of Meiren yak was mainly composed of α-helix and irregular curling.The expression of the PDK4 gene in the liver and lung of 7-day-old Meiren yak was significantly higher than that of other tissues (P<0.05),moderately expressed in muscle,fat,spleen,and testes,and low in the kidney and heart.As the yak aged,the expression of the PDK4 gene in the adipose tissue of 30 months old Meiren yak increased and was significantly higher than that in the fat of 7-day-old Meiren yak(P<0.05).The above results suggest that the PDK4 gene may be involved in fat regulation during the growth process of Meiren yak and play an essential role in the fat deposition process.

To explore the effects of different farming modes on the growth and development,photosynthesis,leaf structure and yield of maize,and to provide a theoretical basis for optimizing the cultivation measures and creating efficient planting patterns of maize in Hexi oasis irrigation area.Two tillage methods,no-tillage(NT)and conventional tillage(CT),and three planting patterns,wheat-maize intercropping(W/M),winter rapeseed-maize rotation after wheat(W-G→M),and wheat-maize rotation(W-M),were set up in the experiment,with a total of 6 treatments.The results showed that compared with CT,the plant height,stem diameter and leaf area of NT maize increased by 6.83%,4.10% and 3.97%,respectively.The dry matter quality of intercropping maize was higher than that of rotation,but the difference was not significant.The leaf pigments increased first and then decreased with the growth period,which showed that NT chlorophyll a,b and carotenoids were 11.93%,22.41% and 13.43% higher than CT,respectively,and the difference was significant.The net photosynthetic rate(Pn)and intercellular CO₂ concentration(Ci)of NT leaves were 9.17% and 3.81% higher than those of CT.The stomatal conductance(Gs)and transpiration rate(Tr)of CT treatment were 9.95% and 1.48% higher than those of NT.The leaf structure of NT maize was better,the mesophyll cells were more and arranged in order,the vascular bundles were clearly visible,the garland structure was larger,the palisade tissue and sponge tissue were rich,and the leaf thickness of NT was 2.51% thicker than that of CT,and the difference was significant.The yield of NT maize increased by 8.02% compared with CT,and the yield benefit of intercropping was greater than that of rotation(LER>1).This study found that the growth and development,leaf structure and yield of no-tillage maize were better than those of traditional tillage,and wheat intercropping maize could be promoted as the main farming mode in this area.

The aim was to screen key differentially expressed genes and signaling pathways and explore the molecular regulatory mechanisms of chicken skin follicle traits through transcriptomic analysis of the dorsal skin follicle tissues of Chongren partridge chicken.Total RNA of skin hair follicle tissue was extracted by TRIzol method,and transcriptome high-throughput sequencing(RNA-Seq)was performed by Illumina platform.Finally,the transcriptome sequencing was validated by Real-time fluorescence quantitative PCR.The results were as follows:A total of 3 856 differentially expressed genes were screened,and 971 differentially expressed genes were screened in the cockerel(BGB vs YGB)comparative combinations,of these,274 up-regulated and 697 down-regulated.There were 3 529 differentially expressed genes in comparative combinations of hen(BMB vs YMB),of these,1 477 up-regulated and 2 052 down-regulated.Three highly expressed genes,KRT75,KRT6A,and KRT14,associated with skin follicle traits,were screened among the common differentially expressed genes in roosters and hens.In the significant enrichment of GO functions,516 GO terms were enriched in the BGB vs YGB and 1 020 GO terms were enriched in the BMB vs YMB.Genes were mainly focused on activities involved in cell adhesion and cell activation.Significantly enriched multiple KEGG pathways,with 8 significant KEGG pathways enriched in the BGB vs YGB and 20 significant KEGG pathways enriched in the BMB vs YMB.Three key pathways,Wnt signaling pathway,neuroactive ligand-receptor regulatory pathway and TGF-β regulatory pathway,were screened.Fluorescence quantification showed that the expression of five randomly selected differential genes in skin hair follicles was consistent with the trend towards RNA-Seq.In summary,the screened KRT75,KRT6A,and KRT14 genes may affect chicken skin follicle traits;the neuroactive ligand-receptor regulatory pathway,the Wnt signaling pathway,and the TGF-β signaling pathway are important pathways that regulate follicular traits in chicken skin.

In order to clarify the impact of irrigation amount on maize production,using Zhengdan 958 as the main material and the local average reference crop water demand(ET0)was used as the basis of irrigation volume,a study was conducted under rain-protection shed in 2020 and 2021,which included eight irrigation treatments:T1,T2,T3,T4,T5,T6,T7 and T8 representing 60% ET0,80% ET0,100% ET0,120% ET0,140% ET0,160% ET0,180% ET0 and 200% ET0,respectively.The study aimed at analysing the effects of irrigation on yield and its components and plant physicochemical properties.The results indicated that the yield and its components,ear length,grain dry matter distribution ratio,free water content,plant height,ear position height,ear height coefficient,photosynthetic potential and seed setting rate of maize showed an increasing trend when irrigation was added.Specifically, the yield and grain number per ear increased significantly while the material distribution ratio decreased in leaf and corn cob.There was a small change rate of chlorophyll and water potential of maize leaves,no significant difference in plant height and ear position height among treatments and stable ear height coefficient at VT and R6 stages under high water treatment(T5—T8).Comparing with T3,a decrease of 20% and 40% in irrigation volume could cause a significant decrease in yield,but the lower yield increase rate appeared when irrigation volume had the same increase rate as the decrease rate. The seed setting rate also showed the same variation under the same condition. However,the water supply efficiency decreased with the increase of irrigation volume and declined more than 20% when the irrigation volume exceeded T6.Correlation analysis showed that correlation between maize yield and leaf chlorophyll content,water potential at anthesis and water content was not significant,but there was a significant or extremely significant correlation with other factors.Therefore,maize yield could be raised by improving ear traits,grain dry matter distribution ratio and physical and chemical properties of plant development through irrigation,an increased irrigation amount by 40%—60% on the basis of the average water demand should be suggested for maximizing the yield and efficiency during the growth period in the region.

This study aimed to investigate the function of the anthocyanidin glycosyltransferase gene(UFGT)in the anthocyanin biosynthesis in common buckwheat.Guihong 1 was used as the experimental material.A UFGT gene,named FeUFGT1,was obtained by homologous alignment analysis,and cloned followed by performing on bioinformatics analysis.Overexpression vector of this gene was constructed,then transferred into Arabidopsis thaliana anthocyanin 3-O-glycosyltransferase mutant atugt78d2 to assess its function.The results revealed that the open reading frame of FeUFGT1 was 1 404 bp and encoded 467 amino acid residues with speculated molecular weight of 51.46 ku and theoretical isoelectric point of 4.97.FeUFGT1 protein contained GT-B type glycosyltransferase family domain and a PSPG-box,specific one of the plant UGT family at the C-terminus.The phylogenetic analysis showed that FeUFGT1 protein was closely related to the 3-O-glucosyltransferase of Siraitia grosvenorii UGT74AC1.The expression level of FeUFGT1 in the white-flower petals of Fengtian 1 was 3.7-fold as high as that in the red-flower petals of Guihong 1 with a significant difference.Mutant recovery experiment showed that FeUFGT1 could restore the phenotype of the Arabidopsis mutant which lacked anthocyanin accumulation.

To explore the function of the TCP gene family in the growth and development of Jasminum sambac,bioinformatics methods were used to identify and analyze the whole genome of J.sambac TCP(JsTCP)gene family.Moreover,the expression levels of TCP gene family in different stages of flower development and pollen-stigma interaction were analyzed.A total of 27 TCP gene family members were identified from the whole genome of J.sambac,named JsTCP1—JsTCP25.The protein contained 208—539 amino acid residues,with molecular weight of 22.95—56.96 ku and isoelectric point of 5.70—9.97,and all of them were unstable hydrophilic proteins.The subcellular localization prediction showed that all JsTCPs were located in the nucleus.JsTCPs were unevenly distributed on 10 chromosomes.Gene structure analysis showed that JsTCPs had 1—5 exons and 0—4 introns.Protein conserved motifs and phylogenetic analysis showed that JsTCPs contained conserved TCP domain,and they were divided into two subclasses:Class Ⅰ and Class Ⅱ.Promoter of JsTCPs contained cis-acting elements related to plant hormone response,stress response,and growth and development.The expression pattern analysis showed that 24 JsTCPs were expressed at different stages of flower development,and 22 genes were expressed in pollen-stigma interactions.In conclusion,27 TCP gene family members were identified from J.sambac,and it found that they expressed specifically at different stages of flower development and different stages after pollination.

In view of the low conversion rate of artificial domestication and survival rate of seedling cultivation of Silurus lanzhouensis,in order to use gene editing technology to create and cultivate new germplasm of S.lanzhouensis that was easy to domesticate,the effects of neuropeptide Y gene(NPY) of S.lanzhouensis were studied.The results showed that the full cDNA sequence of the NPY gene was 775 bp in length,with an open reading frame(ORF)of 288 bp,which encoded 95 amino acids and contained a transmembrane helix and a signal peptide binding site for pancreatic hormone recognition.The NPY protein sequence of S.lanzhouensis had the highest consistency with that of Silurus meridionalis,which was 98.95%.The Real-time Fluorescent Quantitative PCR results showed that NPY mRNA expression was the highest in brain tissue,followed by the hypothalamus,and the lowest in muscle.The expression level of NPY mRNA in male tissues of different ages was higher than that in female tissues;and the expression of NPY mRNA decreased with age in both sexes.Localization of brain cells by in situ hybridization found that NPY was mainly expressed in the cytoplasm,and abundant expression was found in the preoptic area,tectum,pituitary,the lentiform nucleus of cerebellum and the caudate nucleus of the medulla oblongata in different parts of the brain.Further verification of starvation compensation showed that the expression of NPY in brain tissue increased significantly after starvation,and decreased significantly after feeding.The above results showed that NPY gene might play an important role in the growth and feeding process of S.lanzhouensis.It could be used as an important verification of the subsequent functional gene editing and the creation of new germplasm,and had important theoretical significance and technical support for accelerating the breeding of new varieties of S.lanzhouensis.

In order to investigate the reasons for the differences in plant height between SSSL-B50 and HJX74,an F₂ population was constructed by backcrossing SSSL-B50 with HJX74 for genetic analysis and gene mapping of the plant.The genetic analysis showed that F₁ plants exhibited tall plant height.And the separation ratio within F₂ was 3:1(χ²=0.18<3.84)between the tall plant individuals and the semi-dwarf ones,which indicated that the tall plant height of SSSL-B50 was a dominant trait.Based on the linkage analysis between the marker genotypes and phenotypes of plant height in the F₂ population,the gene controlling tall plant height was mapped between markers S18 and X161 within the 38.38—39.07 Mb interval of the substituted segment on chromosome 1.Moreover,it was found that the known gene of "Green revolution",SD1,fell to the mapping interval.Through amplification and sequence alignment of SD1 gene in Oryza glumaepatula,SSSL-B50,and HJX74,it showed that a 280 bp deletion in CDS in HJX74 compared with Oryza glumaepatula and SSSL-B50.The qRT-PCR results showed that the expression level of SD1 was significantly higher in the second,third,and fourth sections of SSSL-B50 stem than in HJX74.The comparison of the SD1 gene sequence of Oryza glumaepatula with the previous reported results showed that the base substitutions occurred at two sites (E100G, Q339R) in the encoded amino acid.The results revealed that the tall plant height of SSSL-B50 was controlled by SD1.Furthermore,a new allelic type of SD1, SD1^Glu,was identified in Oryza glumaepatula.

Calmodulin-like proteins(CMLs),one of the Ca²⁺ receptors in plants,are involved in the process of plant growth and development,as well as adaptation to environmental changes.To understand the sequence characteristics of garlic CMLs and their responses to osmotic stress,AsCML15 and AsCML42 genes from garlic variety Cangshan siliuban were cloned,and their expression patterns under drought and salt stress conditions were determined.The results showed that the open reading frame of AsCML15 and AsCML42 genes were 498 and 543 bp in length,respectively,encoding 165 and 180 amino acid residues.AsCML15 and AsCML42 harbored four and three EF-hand domains,respectively.AsCML42 was closer to Arabidopsis AtCML42 and AtCML43 in evolutionary relationship,whereas AsCML15 was more closely related to Arabidopsis AtCML15 and AtCML16.Real-time Quantitative PCR technology showed that AsCML15 and AsCML42 were expressed in bulbs,leaves,and roots,and these two genes can be induced by 200 mmol/L NaCl and 15% PEG6000.The AsCML15 and AsCML42 genes may be involved in the process of garlic resisting salt and drought stress,and their biological functions can be further identified.

To explore the effects of fertilizer reduction and organic fertilizer application on physicochemical properties and microbial community structure of tobacco-planting soil,and provide theoretical reference for fertilizer reduction and rational application of organic fertilizer in tobacco production.Using conventional fertilization without organic fertilizer as control(CK),Illumina high-throughput sequencing technology combined with bioinformatics,the changes of soil physical and chemical properties and microbial community structure under different treatments such as 10% reduction of fertilizer(T1),10% reduction of fertilizer+sesame cake fertilizer(T2),10% reduction of fertilizer + humic acid fertilizer(T3),and 10% reduction of fertilizer + sesame cake fertilizer + humic acid fertilizer(T4)were analyzed.The results showed that compared with CK,soil nutrient content and soil enzyme activity decreased under T1 treatment,and soil physical properties were slightly improved.Combined with organic fertilizer,soil nutrient and physical properties were further improved,bulk density decreased,moisture content and porosity increased,and the contents of available phosphorus and available potassium under T2 and T4 treatment were significantly higher than those under CK and T1 treatment.The enzyme activity of soil treated with organic fertilizer increased significantly.Combined application of organic fertilizer increased the bacteria and fungi in tobacco-growing soil,among which the dominant bacteria were Proteobacteria and Actinobacteriota.Followed by Firmicutes,Bacteroidota,Chloroflexi and Acidobacteriota.The dominant fungal groups are Ascomycota,Anthophyta,Mortierellomycota,Chlorophyta,Ciliophora and Basidiomycota.The Alpha diversity index showed that the reduction of fertilizer decreased the richness of microbial community,but the combined application of organic fertilizer increased the diversity index of bacterial and fungal communities,and the bacterial community richness increased more significantly.RDA analysis showed that the important soil physicochemical factors affecting soil microbial community structure and diversity included organic matter,available potassium,alkali-hydrolyzed nitrogen,available phosphorus and soil physical properties,etc.Soil physicochemical factors had a greater impact on bacterial community structure.In conclusion,under the condition of fertilizer reduction,organic fertilizer combined with organic fertilizer can significantly improve soil nutrients,physical properties,soil enzyme activity and microflora structure,especially when combined with sesame cake fertilizer and humic acid fertilizer treatment.

Understanding the variation and population structure of the foxtail millet blast provides a theoretical basis for future cultivation of resistant varieties and control of the disease.Twenty pairs of SRAP primers were used for PCR amplification of strains of P.oryzae collected from 9 regions.NTSYSpc-2.11F software was used for data analysis,UPGMA method was used for cluster analysis,and Popgene 32 software was used to calculate the genetic diversity index among populations.The results showed that 8 pairs of primers were selected with good polymorphism.A total of 1 728 discernible bands were amplified by 8 pairs of primers,1 492 of which showed polymorphism,and polymorphism rate was 86.34%.The cluster analysis of 90 strains of P.oryzae showed that the similarity coefficient was between 0.77-0.85.All strains were divided into 27 genetic lineages (L1-L27) when the genetic similarity coefficient was 0.802.Among them,L1 was the absolute dominant group,containing 29 strains of P.oryzae,which from Shandong,Hebei,Shanxi,Henan and Liaoning Province,accounting for 32.22% of the total strains.According to the calculation and analysis of Popgene 32 software,Nei's genetic diversity index (H) and Shannon's information index (I) of foxtail millet blast in 9 regions ranged from 0.141 4 to 0.288 1,and 0.196 0 to 0.441 6,respectively.Nei's genetic diversity index and Shannon's information index in the summer sowing region of Hebei Province were the highest,with the richest genetic diversity,while Hainan population had the lowest genetic diversity.Compared with different regions of the populations,the genetic relationship between the Jilin population and the Hainan population was the farthest,while the genetic relationship between the Hebei summer sowing millet region population and the Hebei spring sowing millet region population was the closest.It can be seen that the genetic diversity of P.oryzae in different regions was rich,and there was genetic differentiation among strains,but there was no significant correlation between genetic differentiation and geographical origin.

MYB transcription factors play an important role in the growth and development of cotton,and GhMYB42 is one of the transcription factors of the MYB family,which also has certain research value,so this study cloned the coding sequence of GhMYB42 gene from cotton and constructed a prokaryotic expression vector.The nucleotide sequence and amino acid sequence of GhMYB42 were analysed using bioinformatics methods, and the coding sequence of the GhMYB42 gene was constructed into the prokaryotic expression vector pGEX-4T-1 by the Gataway BP and LR reactions, and the optimal conditions for the induction of the protein by IPTG were determined by setting up different IPTG-inducing conditions, and finally the recombinant protein was identified by Western Blot.The results showed that the full-length sequence of GhMYB42(XP_016732693.1) was 1 508 bp,the coding region length was 792 bp,the coding area was 263 amino acids,the predicted molecular weight was about 29.534 ku,and the isoelectric point was 5.18.Amino acid sequence comparison analysis revealed that the sequence similarity of the MYB transcription factors was 80.62%, and that the N-terminus of the GhMYB42 protein contained two tandem SANT structural domains, making it an R2R3 transcription factor.Phylogenetic tree analysis showed that the MYB42 protein in Gossypium hirsutum had the highest homology with another MYB protein (XP_012439547.1) in Gossypium hirsutum and was on one branch.Since the results of each experimental gradient were not significantly different during protein induction,it chose the conditions for IPTG to be 0.2 mmol/L at a final concentration of 0.2 mmol/L,a temperature of 37 ℃ for 3 h,and a temperature and time for protein solubility to be induced at 37 ℃ for 3 h.The Western Blot results showed that the size of the recombinant protein was correct,and finally the GhMYB42 recombinant protein with a size of 55.54 ku was successfully obtained,and it will purify the recombinant protein and further study the function of the transcription factor GhMYB42.

The WRKY transcription factor family plays an important role in regulating abiotic stress.To systematically analyze the sequence characteristics and expression patterns of NtWRKY11 gene in tobacco,and explore the response mechanism of NtWRKY11 under abiotic stress such as low temperature and drought,the full-length cDNA sequence of NtWRKY11 gene was amplified by PCR using common tobacco cDNA as a template.The basic properties of NtWRKY11 protein were analyzed by bioinformatics software and the subcellular localization of NtWRKY11 protein was studied by constructing a plant expression vector.The expression of NtWRKY11 gene was detected by qRT-PCR technology in different tissues at the full flowering stage and under different abiotic stresses.The results showed that the full-length cDNA of NtWRKY11 gene in tobacco was 999 bp,encoding 332 amino acids,and it shared 54.23% similarity with ramie BnWRKY11.Its promoter region contained three kinds of cis-acting elements(one MBS,one MYB and three ARBE),which probably worked together to enhance the drought resistance of plants.It also contained three salicylic acid-responsive cis-acting elements(TCA-element),which could improve the low temperature tolerance of plants.Subcellular localization results indicated that the NtWRKY11 protein was located in the nucleus.The expression analysis of NtWRKY11 in different tissues at the full flowering stage showed that NtWRKY11 was highly expressed in old leaves,significantly higher than in roots.However,the expression in flowers was significantly lower than that in roots,and there was no significant difference between stems,new leaves and roots. The expression analysis under abiotic stress showed that the relative expression of the gene was significantly higher than that of normal (CK).The relative expression level under high salt stress was not significantly different from CK,while the relative expression level under high temperature stress was significantly lower than CK.All in all,NtWRKY11 is highly expressed in old leaves,and its expression level is enhanced under abiotic stresses such as drought stress and low temperature stress,indicating that this gene acts as a forward transcription factor to regulate drought and low temperature stress.

The NAC(NAM,ATAF1/2,CUC2)gene is an important regulatory factor in the stress signal transduction network.Cloning the NAC gene in castor,studying its molecular characteristics and expression characteristics,aiming to provide data support for the potential function of the castor NAC gene.The RcNAC100-like gene of Tongbi No.5 was cloned by RT-PCR technology,and its molecular characteristics were analyzed,including bioinformatics,subcellular localization,expression patterns,and transcription activation domain analysis.The results showed that the full length of RcNAC100-like gene cDNA was 1 244 bp,including a 1 086 bp coding sequence(CDS),encoding 361 amino acids.The protein had more irregular coil and α-helix structures,and was a hydrophilic,non-secretory protein.Phylogenetic analysis showed that the RcNAC100-like protein was most closely related to NAC proteins in Manihot esculenta and Hevea brasiliensis,with highly similar motif composition and positioning.The subcellular localization of RcNAC100-like protein was consistent with the predicted results,located in the nucleus.The predicted cis-acting elements in the RcNAC100-like promoter region indicate the presence of multiple environmental response and growth-related elements.Expression pattern analysis showed that the RcNAC100-like gene had tissue-specific expression,with the highest relative expression level in the roots.Additionally,the gene could respond rapidly to adverse environments(drought,salt,cold,and ABA stress)and actively express,indicating that the RcNAC100-like gene might be a key gene in the castor's response to stress.Transcription activation assay results showed that the RcNAC100-like transcription factor has transcriptional activation activity in yeast.In summary,the RcNAC100-like gene may play an important role in the castor's resistance to adversity.

In order to provide theoretical basis for straw fertilizing high-yield field and improving saline-alkali field in the process of continuous high yield and stable yield of spring corn in Inner Mongolia plain irrigation area.This study set up a positioning test of corn straw returning to the field for 1 to 4 years (HT1—HT4), taking the straw not returning to the field as the control (CK). Soil organic carbon content, total nitrogen content, available nitrogen content, available phosphorus content, cation exchange capacity, pH value and acid-base buffer curve were measured before spring sowing and during harvest. The results showed that, the relative change rates of soil organic carbon content, total nitrogen content, available nitrogen content, available phosphorus content and cation exchange capacity were 1.34%—3.62%, 0.20%—1.51%, -0.11%—0.78%, 0.89%—6.36% and 0.09%—0.41% during harvest period of HT1—HT4 compared with that before spring sowing. The relative change rates of soil organic carbon content, total nitrogen content, available nitrogen content, available phosphorus content and cation exchange capacity were 1.57%, -0.02%, -0.45%, -0.15% and -0.05% in CK harvest period compared with that before spring sowing. Soil pH value of HT2, HT3 and HT4 was significantly lower than that of CK. The buffer capacity of soil to alkali was HT4>HT3>HT2>HT1>CK. In summary, with the increase of straw returning years, soil carbon sequestration capacity, fertilizer retention capacity and buffering performance increase, and the ability to effectively resist drastic changes in soil pH value caused by fertilizer application and other factors was enhanced, soil quality was significantly improved by straw returning to field for fertilizer cultivation and soil improvement measures.

WRKY transcription factors play an important role in the regulation of plant growth and defence responses.In order to explore the function of WRKY gene family in chestnut under drought resistance,the chestnut WRKY gene family members were identified and the physical and chemical properties,phylogeny and structure of the encoded proteins were analyzed.Transcriptome sequencing and qRT-PCR were used to analyze the expression characteristics of WRKY gene family members under drought stress.The results showed that 65 WRKY gene family members were identified and divided into three groups:Ⅰ,Ⅱ and Ⅲ,among which group Ⅱ was divided into five subgroups according to its structure and phylogenetic relationship.Analysis of WRKY gene structure and conserved motif showed that the number of exons ranged from 1 to 7,and the number of exons and motif distribution in the same subgroup were similar.The conserved domain of WRKY gene had certain variation,including the deletion of WRKY heptapeptide domain,the deletion of zinc finger structure,and the heptapeptide domain was mutated into WRKYGKK,WRKYGRK and WRKYGRK.Transcriptome sequencing data showed that 4 WRKY family members were not expressed in the samples,and 49 WRKY family members were up-regulated under drought stress,while 11 WRKY family members were down-regulated under drought stress.The above results showed that,chestnut WRKY genes played an important role in response to drought stress.

Basic leucine zipper(bZIP)transcription factor protein is a kind of transcription factor with conservative structure and function in animals,plants and microorganisms.In order to clarify the function and mechanism of bZIP transcription factor in plant pathogenic fungi,and further determine its relationship with the growth,development and pathogenicity of the pathogen,the StbZIP9 gene was cloned from Setosphaeria turcica 01-23(GenBank No.XM _ 008032179.1).StbZIP9 is a member of the bZIP transcription factor family.The analysis of the gene structure and protein characteristics showed that the DNA sequence was 788 bp in length,with an open reading frame of 726 bp,encoding 241 amino acids.The encoded protein contained a highly conserved homologous domain BRLZ in fungi.The RNA-seq data of the gene during the growth and development of the pathogen and the process of infecting the host were analyzed.It was found that the expression level of StbZIP9 was 2 to 4 times higher than that in the appressorium and germ tube period compared with the mycelium period.After 24,72 h of infection of maize leaves,the gene expression increased from scratch and continued to increase,indicating that StbZIP9 was associated with appressorium development and germ tube formation and played an important role in the process of pathogen infecting host cells.Further,bioinformatics techniques were used to predict its binding conserved motifs and regulatory target genes.The binding motif was NNTWACGTNN,including the bZIP transcription factor recognition core sequence ACGT,and the downstream target genes of StbZIP9 were predicted according to the sequence.Combined with the expression pattern analysis using the RNA-seq data,four downstream target genes(protein IDs in the JGI database were :132893,163024,162798,40466)were obtained,and the functional annotation table was obtained.The functional annotation revealed its involvement in many biological processes, such as polymerization and transport of cell wall components, host infection, and spore dormancy. It will provide the basis for further elucidation of the molecular mechanism involved in the regulation of pathogen infection.

In order to improve the selection efficiency of melon materials with different pulp firmness types and accelerate developing varieties.The F₂ and BC₁F₁ populations were constructed with crisp melon materials 19A21,19A75 and soft melon materials N84,20S66 as parents.According to sensory testing statistics,the theoretical values of 3∶1 and 1∶1 for crispy and soft materials were consistent,indicating that the melon pulp firmness was controlled by a single gene,and crisp was dominant to soft.The changes of ACO activity and expression pattern of CmACO1 with different firmness types were analyzed with 4 materials.The results showed that the ACO activity of soft melon appeared at a peak during fruit development,but the crispy melon did not appear.The expression of CmACO1 in soft melon was significantly higher than that in crispy melon.This gene may be involved in regulating pulp firmness.According to the difference of insertion and deletion sites of CmACO1 in different materials,the InDel-Pf marker was developed.The InDel marker was used to detect the genotypes of 32 melon materials.Among them,crisp melon showed deletion band type,soft melon showed non-deletion band type,the marker polymorphism and pulp firmness were co-separated.Two F₂ populations were used to validate the InDel-Pf,the genotype and phenotype coincidence rates were 95.3% and 98.1%.The results showed that InDel-Pf marker had high accuracy in the actual identification of melon pulp firmness,which could effectively improve the efficiency of breeding selection and shorten the breeding cycle of improved varieties.

The aim of this trial was to investigate the effect of different levels of NDF on the expression of genes related to growth axis and duodenal growth in lambs,in order to provide a reference for the appropriate NDF level for lamb starter.Thirty-six male Hu lambs were selected for the trial and randomly divided into 15% and 25% NDF level groups.At 56 days of age,six lambs from each group were selected for slaughter,weighed for tissue and organ,and the expression of relevant genes was measured.The results showed that there was no significant effect(P>0.05)of NDF levels in the starter diet on tissue and organ mass and index;in the 15% group of lambs,hypothalamic growth hormone releasing hormone(GHRH),hepatic insulin-like growth factor-1(IGF-1),duodenal IGF-1 and IGF-1 receptor(IGF-1R)expressions were significantly higher in the 25% group(P<0.05);the expression of pituitary growth hormone(GH)and growth hormone receptor(GHR) in the duodenum were significantly higher in the 25% group than in the 15% group(P<0.01).The average daily gain of lambs was strongly negatively correlated with hypothalamic GHRH expression(P<0.01),significantly negatively correlated with hepatic GHR and duodenal IGF-1 expression(P<0.05),and significantly positively correlated with pituitary GH and duodenal GHR expression(P<0.05);pituitary GH was significantly or highly significantly negatively correlated with hepatic GHR,IGF-1,duodenal IGF-1 and IGF-1R expression(P<0.05 or P<0.01),and strongly significantly positively correlated with duodenal GHR expression(P<0.01);hepatic GHR was strongly significantly positively correlated with hepatic IGF-1,duodenal IGF-1 expression(P<0.01),and strongly significantly negatively correlated with duodenal GHR expression(P<0.01).In conclusion,the expression of genes related to GH-GHR levels in the growth axis of lambs is enhanced by feeding 25% NDF level open feed compared to the 15% level group.

To improve seedling blast resistance of rice cytoplasmic male sterile(CMS)line Taonong1A and its maintainer Taonong1B,the Chinese local rice variety Gumei 4 was used as the donor parent harboring the broad-spectrum and durable blast resistance gene Pigm,and the Pigm-tightly-linked codominant marker T9E3 was developed and employed in marker-assisted selection(MAS)breeding practice.T9E3 showed clear and stable polymorphism between the donor and acceptor parents,amplifying a 926 bp DNA band from Gumei 4 genome,while the PCR product from either genome of Taonong1B or Taonong1A was about 2 000 bp.Greenhouse inoculation assay was performed using thirty-two Magnaporthe oryzae isolates collected from different rice areas,Gumei 4 and the improved rice lines showed much higher resistance frequency(90.63%)than that of two acceptor parents Taonong1B and Taonong1A(9.38%).Accordingly,Gumei 4 and the improved rice lines showed high-level seedling blast resistance(0 grade)but highly susceptible(8 grade)for two acceptor parents in natural nursery.Three blast resistant CMS pure lines(Taonong1A-Pigm-1—Taonong1A-Pigm-3)and the corresponding resistant maintainers(Taonong1B-Pigm-1—Taonong1B-Pigm-3)were preliminarily bred through MAS breeding,and one pair of them,Taonong1A-Pigm-2/Taonong1B-Pigm-2,was screened for potential application duo to the high-level blast resistance and elite sterility,agronomic and yield traits.Taonong1A-Pigm-2 showed complete male sterility with nearly 60% typical abortive pollens and 40% round abortive pollens,higher stigma exsertion rate(71.1%)and lower enclosed spikelet rate(33.6%),Taonong1B-Pigm-2 exhibited excellent agronomic and yield trait indexes,such as moderate sowing-heading duration(69 days),plant height(76.2 cm),main panicle length(28.1 cm),panicles per plant(15.8),spikelets per panicle(115.3)and 1000-grain weight(27.7 g)。New parent materials were created for the application of cytoplasmic male sterile(CMS)line hybrid rice breeding,and the comprehensive improvement of seedling blast resistance and characters of Taonong1A and Taonong1B were realized.

Breeding resistant varieties is the most economical and feasible method to control wheat leaf rust.In order to further explore the resistance genes,50 wheat varieties were selected in this study,including wheat varieties from Shandong,Henan,Hebei,Shandong and other 8 provinces.Firstly,16 physiological races of leaf rust fungus (THFS,TGTS,THJS,FHKT,FGJN,KHKS,FCJQ,RFKS,THFM,MHGT,KHGS,KBGT,FHGT,PHHT,FHJT,FCJT) were inoculated to 36 vector cultivars containing known leaf rust resistance genes,and 50 wheat varieties tested during the seedling stage.Due to the different virulence of each strain,specific molecular markers closely linked to known disease resistance genes can be combined and analyzed based on the differences in phenotype,and it can be inferred that 50 wheat materials may contain leaf rust resistance genes.The leaf rust resistance genes were identified by genetic deduction,molecular markers and pedigree analysis.The results showed that a total of 9 known resistance genes (Lr1,Lr2c,Lr10,Lr16,Lr26,Lr34,Lr37,Lr45 and Lr46) and a few unknown genes were detected in 50 cultivars.There were 22 varieties including Zimai 12 that contained the Lr1 gene;there were 10 varieties including Lumai 14 that contained the Lr2c gene;Laizhou 9361 only contained the Lr10 gene;there were 25 varieties including Kenong 199 that contained the Lr16 gene;there were 15 varieties including Xuzhou 24 that contained the Lr26 gene;Baomai 3 and Jingdong 8 contained the Lr34 gene;The Lr37 gene was contained in Zimai 12,He 0927,and He 9946;there were 11 varieties including Lianmai 2 that contained the Lr45 gene;there were 38 varieties including Shannong 19 that contained the Lr46 gene.

In order to explore the effects of melatonin on the growth and development of adzuki bean seedlings under salt stress,a phytotron experiment was conducted with Baohong 876 as the material and four treatments:CK(Clear water),S(60 mmol/L NaCl),MT(50 μmol/L melatonin)and S-MT(60 mmol/L NaCl and 50 μmol/L melatonin).The growth indexes,photosynthetic indexes,mineral ion contents and antioxidant enzyme activities of adzuki seedlings were analyzed using classical physicochemical determination.The results showed as follows:under salt stress,compared with CK treatment,the growth of adzuki bean seedlings was significantly inhibited,chlorophyll content,net photosynthetic rate(Pn),transpiration rate(Tr),stomatal conductivity(Gs)and intercellular CO₂ concentration(Ci)were significantly decreased;the contents of Na⁺ in leaves,stems were significantly increased.Meantime,the content of Ca²⁺ in leaves was significantly increased while the content of Mg²⁺ was significantly decreased,and the ratio of Na⁺/K⁺ was significantly increased.The contents of MDA and H₂O₂ increased significantly,which damaged the selective permeability of membrane lipid.Compared with S treatment,the plant height,leaf area,biomass,and total root length of adzuki bean seedlings were significantly increased under S-MT condition.Meantime,chlorophyll content,leaf Pn,Tr,and Ci were increased.The contents of Na⁺ and K⁺ in leaves,stems were significantly decreased,the content of Mg²⁺ in leaves was significantly increased and Ca²⁺ was significantly decreased,and the Na⁺/K⁺ ratio was decreased.The contents of MDA, {\mathrm{O}}_2^{-} and H₂O₂ decreased.The levels of SOD,POD and CAT activity in leaves and roots were increased.Under normal conditions,the application of melatonin could promote the growth of adzuki bean seedlings,improve the photosynthetic capacity of seedlings,and increase the content of soluble protein.Spearman correlation analysis was performed on 15 representative indicators,and it was found that total root length,chlorophyll content,net photosynthetic rate,and antioxidant enzyme activities were significantly or extremely significantly negatively correlated with Na⁺/K⁺ ratio and ROS.In summary,under salt stress,melatonin can directly and indirectly reduce H₂O₂ content and maintain the stability of membrane lipid permeability by activating the antioxidant enzyme activity of adzuki bean seedlings,thus maintaining the Na⁺/K⁺ balance in plants.Meanwhile,H₂O₂ and Ca²⁺,as signaling molecules,have a synergistic effect in the regulation process of melatonin regulating intracellular ion balance,thereby improving the salt tolerance of plants.

Effective spike number is an important component of castor yield.In order to reveal the genetic basis and mining candidate genes of effective spike number,the QTLs conferring effective spike number per plant(ESNPP)and number of effective primary branch spike(EPBSN)were mapped by genotyping with SSR primers and by the 2 detection methods(CIM and ICIM-ADD)in the populations F₂ and BC₁ (F₁×P₂)derived from 2 parents with significant phenotypic differences,and QTL remapping was conducted in S₁ population by the same method.A total of 9/5(CIM/ICIM-ADD,the same below) QTLs were identified in the F₂ population,including 3/2 and 6/3 underlying ESNPP and EPBSN,with a phenotypic variation explained(PVE)of 6.70%/11.87% and 25.15%/13.87%,respectively.The mapping results in BC₁ population were generally consistent with those in F₂ population.Among them, qESNPP3.1 and qEPBSN3.1 were stable QTLs with contribution rates close to 10%,and the latter was detected again in S₁ population with a PVE of 13.27%;the stable QTLs were overlappingly located in the marker interval RCM915—RCM950,and together constituted a main-effect QTL cluster underlying the effective spike number.Two candidate genes,Rc-US03g04880 and Rc-US03g05950,were annotated from the marker interval,which were differentially expressed in multiple branching tissues of both parents.The above results provided essential molecular markers for marker-assisted selection of the effective spike number and necessary candidate genes for its genetic and physiologic mechanism study in castor.

The transcription factor BnHY5-2 is associated with plant stress resistance.In order to reveal the response of Brassica napus L.transcription factor BnHY5-2 to salt alkali stress in B.napus,the response of BnHY5-2 to light and salt and alkali was analyzed by transient overexpression,qRT-PCR analysis and subcellular localization.The results revealed that under light conditions,the expression level of the BnHY5-2 gene in B.napus leaves and stems was 29.22 and 3.15 fold higher,respectively,compared to dark conditions.The higher sensitivity to light in leaves suggested that they were the primary site for light signal response.Under light conditions,the expression of BnHY5-2 in leaves and stems was significantly downregulated by 53.1% and 31.0%,respectively,when B.napus was planted in Dalian coastal saline-alkali soil;after applying saline-alkali treatment under dark conditions,the expression of BnHY5-2 was downregulated by 48.2% in the stem,while the difference in expression in the leaves was not significant,indicating organ differences,indicating that the leaves had stricter requirements for light conditions.In B.napus leaves with transient overexpression of BnHY5-2,two out of six genes related to saline-alkali stress(BnNAC32 and BnGS)showed upregulation by 1.25,3.28 fold,respectively,while the other four genes(Bnamy,BnAsp,BnNHX7,BnTPS)were downregulated by 24.8%,25.4%,71.0%,and 82.0%,respectively.Meanwhile,the content of the resistance substance betaine in B.napus increased from 0.256 to 0.573 mg/g,indicating an enhancement by 1.24 fold,suggesting that the overexpression of BnHY5-2 gene could improve the saline-alkali tolerance of B.napus.Subcellular localization results showed that the transcription factor BnHY5-2 was localized in the nucleus and regulates the expression of functional genes.Therefore,BnHY5-2 is not only related to light signaling but also participates in the saline-alkali resistance of Brassica napus L.

By screening differentially expressed genes and metabolites in the green and purple leaves of Brassica napus,it lays a theoretical foundation for the analysis of mechanism of anthocyanin synthesis.Phenotypic observation of GL and PL seedlings showed that PL exhibited a mottled distribution of purple when two true leaves were grown(3 weeks),as the purple gradually deepened during development,it gradually became lighter and eventually disappeared after the bolting stage(>16 weeks),while GL leaves remained green throughout the entire period;transcriptome analysis identified 2 523 co-differentially expressed genes at weeks 6,13,and 16,and these differentially expressed genes were significantly enriched in the anthocyanin synthesis pathway.Twenty-four genes related to anthocyanin synthesis were significantly differentially expressed,including 3 MYBL2,1 C4H,1 F3H,2 F3'H,2 TT8,3 DFR,4 ANS,5 UGT,and 3 TT19.The expression levels of these genes in PL leaves were higher than those in GL at 6 and 13 weeks.Eight differentially expressed genes were selected for qRT-PCR validation and the results were consistent with the trend of transcriptome analysis data.A total of 50 anthocyanins were detected in the anthocyanin-targeted metabolome,of which 29 showed significant difference in accumulation;compared to GL,PL had 16 types of anthocyanin accumulation upregulated and 13 types of anthocyanin accumulation downregulated.The up-regulation of TT8 and its target genes(DFR,ANS,UGT,TT19)in early development(6—13 weeks)promoted the accumulation of cyanidin-based anthocyanins and the decrease of petunian-based anthocyanins in PL.The expression levels of differentially expressed genes and metabolites in the anthocyanin synthesis pathway reached significant levels.

In order to study the mechanism of PSY1 gene in different ripe fruit colors of peppers, Y15016,Y15016-2,SP01,SP02 and Z1 were used as materials,and the functional properties of PSY protein and the expression of PSY1 gene in different fruit color materials of pepper were studied and analyzed by combining some bioinformatics methods.The results showed that the full-length PSY1 gene could be cloned in all five pepper varieties,and there was no difference in sequence.Gene structure analysis showed that the PSY1 gene contained six exons and five introns with a total length of 2 844 bp,and its CDS contained 1 260 bp and encoded 419 amino acids.Sequence alignment and phylogenetic tree analysis showed that the PSY protein of pepper was the closest to the homologous PSY protein of tomato and tobacco of the same family.The results of qRT-PCR analysis showed that the expression level of PSY1 gene in the five materials was lowest in the root tissue and highest in the leaf tissue.Besides,the PSY1 gene was expressed in all the tissues of these materials.The expression level of orange mutant Y15016-2 was higher than that of wild-type Y15016,while that of yellow mutant SP02 was significantly lower than that of wild-type SP01.At different stages of fruit development,the expression of PSY1 gene increased with the development of fruits,except for the decrease in stage Ⅲ,and reached the maximum value at the maturity stage(Ⅳ—Ⅴ)of different fruit color materials.The results of PSY1 gene promoter analysis showed that there was no difference in the sequences in the test materials.The results suggest that the differential expression of PSY1 gene may play an important role in the formation of peppers with different fruit colors.

To clear under the condition of slow release nitrogen fertilizer applying different content of maize development rule of the diurnal change of photosynthetic characteristics and growth,the light response curve was to fit,so as to farming-pastoral zone of spring maize cultivation and nitrogen utilization to provide theoretical basis.This study took Guangde 5 as the research object,and based on the long-term positioning experiment in 2018,the leaves ear position in spinning—tasseling stage of maize were mesured and analyzed under six nitrogen gradient of N 0(N0,CK), 120(N8), 180(N12), 240(N16), 300(N20),360(N24) kg/ha in 2019 and 2020. SPAD value, intercellular carbon dioxide concentration, stomatal conductivity, diurnal variation of net photosynthetic rate, transpiration rate, light response curve and dry matter accumulation rule.The results showed that with the increase of nitrogen fertilizer application, the changes of SPAD value, net photosynthetic rate, transpiration rate and stomatal conduction degree day of maize increased first and then decreased,and N16 treatment was the highest,and intercellular carbon dioxide concentration was the lowest.The changes of net photosynthetic rate,transpiration rate and stomatal conduction degree day showed unimodal curve.The light response curve fitting analysis showed that the maximum net photosynthetic rate of N16 treatment was the highest.In 2019 and 2020,N16 treatments were 37.48%,29.51%,31.85%,18.17%,37.32% and 80.04%,59.73%,50.30%,6.42%,62.51% higher than N0,N8,N12,N20 and N24 treatments,respectively.Dry matter mass accumulation in whole plant and ear were positively correlated with maximum net photosynthetic rate,apparent quantum efficiency and SPAD value.Comprehensive analysis showed that the most suitable nitrogen fertilizer application rate in western Inner Mongolia was 240 kg/ha.

In order to explore the effects of straw and microbial fertilizer on the salt content of soil and crops in saline-alkali soil,the experiment was conducted in Dalat Banner,Inner Mongolia.Four treatments were set up:no straw and no microbial fertilizer(CK),single application of straw(G),single application of microbial fertilizer(F)and combined application of straw and microbial fertilizer(GF).Four crops,oat,sorghum hybrid sudangrass,proso millet and oil sunflower,were planted,respectively.The effects of straw and microbial fertilizer on salt ion concentration and salt content of soil and crops in saline-alkali soil were analyzed.The results showed that compared with CK,straw combined with microbial fertilizer could significantly reduce soil salt content,increase crop biomass and salt ion accumulation in crops,and the soil total salt content in the soil planted with oil sunflower was the lowest at 4 400.41 mg/kg,which was significantly reduced by 13.85% compared with CK.Crop biomass and salt ion accumulation of sorghum hybrid sudangrass treated with straw combined with microbial fertilizer were the largest,and the biomass and salt ion accumulation were increased by 68.32% and 108.28% compared with CK,respectively.In addition,straw combined with microbial fertilizer enhanced the selective absorption ability of each crop for salt ions.Under combined treatment,oat had stronger absorption ability for Mg²⁺,Cl^- and {{\mathrm{SO}}_4}^{2-},which were 50.51%,72.74% and 56.39% higher than CK,respectively.The selective absorption capacity of Ca²⁺ was 76.01% higher than that of CK.Oil sunflower mainly absorbed K⁺,and its content was 12.90% higher than CK.In conclusion,straw combined with microbial fertilizer could significantly improve crop biomass,salt ion accumulation and salt ion selective absorption ability,and could effectively reduce the total salt content of saline-alkali soil.Among the 4 crops,sorghum hybrid sudangrass and oil sunflower perform better.

To investigate the nature and function of PlSPL1 (SPL)gene of Paeonia lactiflora,further elucidate the commonalities and characteristic differences of the PlSPL1 gene in different species,and explore the role of PlSPL1 in the degree of stem straightening in Paeonia lactiflora. The full-length sequence of the PlSPL1 gene was obtained by RACE technology using the stalk of herbaceous peony Hongfeng as the research material,and the structural,physicochemical properties,and phylogenetic relationships of PlSPL1 were analyzed and predicted using bioinformatics software,then,the expression level of PlSPL1 in different developmental stages of herbaceous peony stems was analyzed using qRT-PCR technology,and subcellular localization analysis of the protein was conducted using laser confocal microscopy technology.The results showed that the open reading frame of the PlSPL1 gene was 3 000 bp,encoding 999 amino acids.The protein had a molecular formula of C₄₈₆₉H₇₆₈₂N₁₄₀₆O₁₄₉₇S₄₃,a molecular weight of 111.25 ku,a theoretical isoelectric point of 6.26,encoding a hydrophilic unstable acidic protein,phosphorylation modification predominantly on serine,no signal peptide,with transmembrane structure,and had a secondary structure consisting mainly of random coil.Phylogenetic tree analysis revealed that the PlSPL1 protein was most closely related to peony,followed by a closer relationship with grape;protein sequence comparison analysis revealed that the PlSPL1 protein had a conserved SBP domain which was unique to SPL transcription factor family.The relative expression analysis found that PlSPL1 gradually showed a decreasing trend with stem development,indicating that PlSPL1 was negatively regulating the stem development of herbaceous peony and it was hypothesised that PlSPL1 played important roles in degree of stem straightness;and the subcellular localisation showed that the PlSPL1 protein was localised in the nucleus.The above results indicate that PlSPL1 is participated in the stalk development of herbaceous peony.

Drought stress has a serious effect on the growth and development of maize,which leads to a decrease of maize yield.Purple acid phosphatase is a phospholipase protein involved in many physiological and biochemical functions of plants.In order to further study the role of purple acid phosphatase family genes in the process of stress resistance of maize,this paper explored the response mode of ZmPAP26b gene under drought stress,and Real-time fluorescence Quantitavive analysis was used to analyze the relative gene expression in different maize inbreeding lines under simulated drought conditions;ZmPAP26b(GenBank:NC_050104.1)was cloned from maize,and PAP genes in Zea mays,Arabidopsis thaliana,Oryza sativa L.,Triticum aestivum L.,Sorghum bicolor and Brachypodium distachyon were identified and bioinformatic analysis was performed.Meanwhile,prokaryotic overexpression strains were constructed for functional verification.The results showed that the expression of this gene decreased in drought tolerant materials and increased in drought sensitive materials under drought stress.The CDS length of this gene was 1 431 bp,encoding 476 amino acids.A total of 228 PAP genes were found in six species,divided into 4 subfamilies by phylogenetic analysis.The 19 PAP genes in maize were distributed on 9 chromosomes and had similar conserved domains.Analysis of promoter cis-acting elements showed that they contained elements responding to drought and hormones.Prokaryotic expression experiments showed that the growth of strains containing the recombinant plasmid pET28a-ZmPAP26b was inhibited compared with non-loaded strains under 10% PEG-6000 and 15%PEG-6000 simulated drought stress.In summary,it is speculated that ZmPAP26b is negatively regulated under drought stress.

To improve the applicability of biochar in saline-alkali agroecosystem,the effect and microbial mechanism of modified biochar were studied.In a 2-year field experiment,common biochar(4.5 t/ha),nitrogen-rich modified biochar(7.5 t/ha)and phosphorus-rich modified biochar(15.0 t/ha)were added to investigate their impact on crop grain yield,soil physicochemical properties and soil microbial diversity.It had been observed that the addition of biochar enhanced the quality of saline-alkali soil,with nitrogen-rich modified biochar and phosphorus-rich modified biochar demonstrating more notable effects.Biochar could boost crop yield,improve soil structure and reduce soil bulk density in saline-alkali land.The effects of the three biochar types were not consistent.Among them,the application of 15.0 t/ha phosphorus-rich modified biochar showed favorable responses,with grain yield of(8.92±0.12)t/ha,representing a 110% increase compared to the control group.Biochar affected soil microbial diversity.Common biochar increased soil microbial diversity,whereas phosphorus-rich modified biochar decreased it.With the continuous addition of biochar,soil physical and chemical properties could affect the relationship between soil microorganisms and plant structure,weakening their relationships.In this study,the application of 15.0 t/ha phosphorus-rich modified biochar was recommended to improve saline-alkali agroecosystem.

In order to study the differences in yield and nitrogen utilization,transport and accumulation of different winter wheat cultivar(line)types in the Northern Huanghuai,the yield traits and nitrogen utilization efficiency of 29 and 26 winter wheat cultivars(lines)for testing in the Northern Huanghuai wheat area were investigated and analyzed in 2021—2022 and 2022—2023,respectively.Through cluster analysis,wheat was classified into four types:high-yielding,medium-high-yielding,medium-yielding and low-yielding.Nitrogen content of wheat stems,leaves,spikes,and grains at maturity were measured at anthesis and maturity,respectively,to analyze the relationship between parameters related to nitrogen utilization traits and yield.The results showed that the average yields among high-yielding,medium-high-yielding,medium-yielding and low-yielding varieties(lines)differed significantly from 2021 to 2023,with the number of spikes per hectare of high-yielding varieties significantly higher than that of other types.At the anthesis stage,the nitrogen accumulation and distribution rate of each organ were stems and sheaths>leaves>spikes;at the maturity stage,the nitrogen accumulation of each organ was grains>stems and sheaths>spikes>leaves.The post-anthesis nitrogen accumulation,pre-anthesis nitrogen transport and its contribution to the grain of different varieties of wheat were higher in high-yielding varieties(lines),and the pre-anthesis nitrogen transport and its contribution to the grain were greater than the post-anthesis nitrogen accumulation and its contribution to the grain.Nitrogen utilization efficiency,nitrogen harvest index,nitrogen accumulation at anthesis and nitrogen accumulation at maturity were significantly and positively correlated with wheat grain yield.Therefore,water and fertilizer management can be carried out through the nitrogen uptake and transfer pattern and distribution characteristics of different varieties,or selecting and breeding wheat varieties with high nitrogen utilization efficiency to achieve high-yield and high-efficiency production of wheat.

Obtaining the LaeA gene sequence of Ganoderma lucidum through electronic cloning,analyzing its gene sequence information and preliminarily exploring its regulatory function.This study adopts the method of electronic cloning,using the known LaeA protein sequence of Penicillium citrinum as a template,perform sequence similarity search and alignment(Blast)in the EST database of Ganoderma lucidum and obtain the cDNA sequence of the LaeA gene of Ganoderma lucidum through electronic cloning methods such as sequence splicing,sequence validation and sequence extension.Some characters of amino acids encoded by LaeA gene,including the physical and chemical properties,hydrophobicity/hydrophilicity,subcellular localization,secondary and tertiary structure of protein,and phylogenetic relationship were analyzed by bioinformatics tools.The length of LaeA gene from G.lucidum was 1 134 bp,encoded 378 amino acids.The protein encoded molecular weight of 42.895 3 ku.The protein was an instability protein that was present in the cytoplasm and not secreted to the extracellular.Furthermore,the structure of LaeA protein was mainly composed by 47.88% random coil,33.33%α-helix and 18.78% extension strand,contained SAM binding site,belonged to AdoMet_MTases superfamily proteins.Phylogenetic analysis showed that LaeA was closely related to white rot basidiomycetes such as Trametes versicolor and Dichomitus squalens;the Real-time Fluorescence Quantitative PCR results showed that the expression level of LaeA in Ganoderma lucidum cells during liquid static culture was significantly higher than in oscillatory culture.It is speculated that the LaeA protein as a methyltransferase protein that participates in histone methylation modification,thereby affecting the expression level of gene clusters.

To investigate the influence of green manure cultivation on the carbon and nitrogen content of saline-alkali soil under freshwater leaching,a field experiment was conducted from October 2021 to May 2022.Three treatments were set up:winter fallow(T1),Dongmu 70 rye(T2),and rape(T3).The spatial distribution of soil organic carbon(SOC),nitrate nitrogen($\mathrm{NO}_{3}{ }^{-}-\mathrm{N}$),and ammonium nitrogen($\mathrm{NH}_{4}^{+}-\mathrm{N}$)in the soil and leachate were measured.The results revealed that in the 0—30 cm soil layer,the soil organic carbon content of T1,T2,and T3 increased from 6.20,6.58,7.24 g/kg before leaching to 6.48,7.39,8.06 g/kg after leaching,representing an increase of 4.41%,12.20%,and 11.23%,respectively.After freshwater leaching,the nitrate nitrogen content in the 0—60 cm soil layer of T1 was significantly higher than that of T2 and T3.In the 0—30 cm soil layer,the respective reductions for each treatment were 42.42%,3.85%,and 10.84%.In the 60—90 cm soil layer,the reductions were 1.38%,7.96% and 18.11%.There were no significant differences in ammonium nitrogen content among the different treatments before leaching,but after leaching,the highest ammonium nitrogen content was observed in T2.In conclusion,after leaching irrigation,the soil organic carbon content in different soil layers increased compared to before leaching,while soil nitrogen showed a significant decrease.Analysis of nitrogen content in the soil and leachate indicated that the main nitrogen loss caused by leaching irrigation was in the form of nitrate nitrogen.Compared to winter fallow farmland,the cultivation of rapeseed green manure had a significant effect on increasing soil nitrogen content,while Dongmu 70 green manure was the the most effective in reducing soil nitrogen loss.

GRAS transcription factors exist extensively in plant genomes and play an important role in both biological and abiotic stress in plants.To study the significant role of the response to stress in soybean cyst nematode (SCN).The susceptible and disease-resistant varieties Dongnong L-10 and Heinong 37 were inoculated with SCN 3 physiological subspecies respectively.After magenta staining was used to confirm the success of insect grafting,transcriptome sequencing was performed on plant roots.A total of 20 candidate genes related to SCN were obtained.The secondary and tertiary structures,phosphorylation sites,hydrophilicity and hydrophobicity,promoter information and gene evolution relationship of encoded proteins were analyzed and studied in bioinformatics.mRNA was extracted from different tissue parts of the plant,including roots,stems and leaves,and reverse transcription cDNA was performed for key candidate genes by RT-PCR.The results showed that the genes in this family were closely related to MYB and MYC transcription factors.The protein encoded by the gene was mainly α-helix and random coil,and the secondary structure and tertiary structure were highly consistent.Protein was a soluble protein,the phosphorylation site was mainly serine.RT-PCR showed that there were certain expression levels in different parts of plant tissues,mainly in roots.The GRAS expression level of resistant cultivars was significantly higher than that of susceptible cultivars under the same soybean cyst nematode stress.GRAS transcription factors were identified as SCN-related resistance genes,providing an important source of genes for resistance to stress response.

In order to provide a theoretical basis for rational application of organic fertilizers and ensure the healthy development of soil ecosystems in corn fields,the effects of soil fungal communities under different levels of organic fertilizer application for the composition,diversity,and functional characteristic were studied.In the meantime,a field experiment was used,with a total of 4 treatments were set up,and the high-throughput sequencing and FUNGuild method were used to analyze the soil fungal diversity,community structure and functional groups in different fertilization rates.The results showed that the increased application of organic fertilizer could increase the content of soil organic matter,available phosphorus,available potassium,urease,catalase,etc.,and it was positively correlated with the application amount of organic fertilizer.Applied organic fertilizers could increase the diversity of soil fungal communities and reduce their richness.The dominant fungal communities in the soil under different fertilization rates were Ascomycota,Mortierellomycota,Basidiomycota,and Olpidiomycota,the relative abundance of Ascomycota and Basidiomycota was significantly higher in the treatment with a medium amount of cattle manure than in the control without organic fertilizer;from the perspective of fungal genera,the dominant genera include Fusarium,Humicola,Olpidium,and Microidium.The application of organic fertilizer increased the relative abundance of symbiotic and saprophytic nutrients,and with the increase of organic fertilizer application,the abundance of pathological nutrients showed a decreasing trend;the number of woody saprophytic fungi in the treatment with increased organic fertilizer application was significantly higher than that in the treatment without organic fertilizer application,while the number of plant and animal pathogens was lower than the control.Therefore,it is believed that increasing a certain amount of organic fertilizer application could optimize the soil microbial environment and promote the increase of corn plant yield.

The aim of this study was to understand the structure and function of ferritin light chain(FTL)gene in yak and its expression pattern in eight tissues of different age groups.The coding sequence(CDS)of FTL gene was cloned from the cDNA of liver tissue of adult(36 months old)Meiren yak,and bioinformatics analysis was carried out.The relative expression of FTL gene in heart,liver,spleen,lung,kidney,longissimus dorsi,fat and testis was detected by Real-time fluorescence quantitative(qRT-PCR).The results showed that the CDS region of FTL gene of Meiren yak was 528 bp,encoding 175 amino acids,and had the closest relationship with wild yak and cattle(99.8%).FTL protein was a stable hydrophilic secretory protein without signal peptide and transmembrane structure.It was mainly distributed in the cytoplasm.The secondary structure of the protein was mainly α-helix,which mainly interacted with FTH1,SCARA5,NCOA4 and other proteins.The results of qRT-PCR showed that the expression levels of FTL gene in adult cattle were liver,kidney,testis,spleen,lung,fat,longissimus dorsi and heart,respectively.The expression levels of FTL gene in fat and longissimus dorsi tissues of 7-day-old calves were the highest,and the expression level of FTL gene in liver tissue of adult cattle was the highest.By comparing the two stages,it was found that the expression levels of fat,longissimus dorsi and heart tissues decreased with the increase of age,while the expression levels of lung,spleen,testis,kidney and liver tissues increased with age.

Leaf color mutant is an ideal material for studying C4 photosynthesis pathway and chlorophyll metabolism mechanism.In order to study the molecular mechanism of yellow-green leaf mutation in millet,and lay a foundation for the functional study of yellow-green leaf genes and the molecular mechanism analysis of chlorophyll metabolism,a stable hereditary yellow-green leaf mutant ygl7 was identified in the ethyl methacrylate(EMS)mutant library of Changnong 35.Agronomic traits,photosynthetic pigment content and photosynthetic parameters,and chloroplast ultrastructure observation were carried out on the mutant and wild type.At the same time,genetic analysis of mutant leaf color was performed,primary mapping was performed by BSA method,fine mapping was performed by an F₂ population,and candidate genes were predicted according to functional annotation combined with RNA-Seq.The expression pattern was analyzed by qRT-PCR,and the protein interaction was verified by yeast two-hybrid experiment.The results showed that the leaves of ygl7 were obviously yellow-green at seedling stage and elongation stage,and gradually turned to light green at heading stage compared with the wild type.The chlorophyll content and photosynthetic rate of ygl7 during the whole growth stage were significantly lower than that of the wild type,and the chloroplast structure was abnormal.Genetic analysis showed that ygl7 yellow-green leaf phenotype was controlled by a pair of recessive single genes.Yellow-green leaf gene was located in the 434.9 kb region of chromosome Ⅶ.Candidate gene analysis predicted that Seita.7G290300 encoding protoporphyrin Ⅸ magnesium chelatase Ⅰ was the candidate gene for regulating yellow-green leaf.The results of qRT-PCR showed that Seita.7G290300 was highly expressed in leaves,and the expression of Seita.7G290300 in mutant was lower than that of wild type.The expression levels of genes related to chlorophyll synthesis pathway(CHLD,CHLI)and photosystem(LHCB1,LHCB6)were down-regulated in the mutants.The experiment of yeast two-hybrid showed that SiYGL7 interacted with MORF2.

The physiological and biochemical indices of cold tolerance in common overwintering cultivated rice seedling were studied for providing theoretical basis for the innovation and utilization of overwintering cultivated rice germplasm. A total of 20 overwintering cultivated rice germplasms were selected as experimental materials,altogether five physiological and biochemical indices such as chlorophyll,malondialdehyde and SOD activity in rice seedling were measured and analyzed under both normal temperature(25 ℃)and 4 days of low temperature(4 ℃)stress. The five cold tolerance indices under normal temperature condition displayed extremely significant phenotypic differences among all tested materials from seven different provinces in China. The cold tolerance index such as chlorophyll and soluble sugar content apparently declined after chilling stress,but exhibited an inconsistent rate of decline among all experimental materials,ranged from -36.46% to -90.31%,-70.54% to -94.03%,respectively. While the cold tolerance indices such as malondialdehyde,proline and SOD activity in all tested rice seedlings displayed disparate response to cold chilling and exhibited an inconsistent rate of rising or falling. Among all tested materials,the malondialdehyde content in three tested materials such as Huangnuo 2,Wanjingnuo M99037 and Changnongjing 3 exhibited significantly decline after chilling stress,the remaining seventeen tested materials exhibited a significant rise. The proline content in eight tested materials such as Tiejingdao 4,Zhendao 7 and Jindao 1187 exhibited a significantly rising after being chilling stress,but the remaining twelve tested materials exhibited a significant decline malondialdehyde content value. The SOD activity in Ewandao 15 and Jinjingzasi exhibited an inconspicuous decline after chilling stress,while the SOD activity in the remaining eighteen tested materials exhibiteda significant rise. Only 2 pairs of extremely significant PCC values among the five cold tolerance indices were calculated under the normal temperature environment,while 8 pairs of extremely significant PCC values were calculated after chilling stress. Apparently,the number,strength and direction of correlations among the 5 cold tolerance indices were easily varied after chilling stress. On the whole,the cold tolerance indices in rice seedling under both normal temperature and low temperature exhibited significant difference owing to the different genetic background of overwintering cultivated rice.

In order to reveal the physiological mechanism of winter rye in response to low temperature stress,the physiological changes of osmoregulatory substances and antioxidant enzyme activity in the leaves and tillering nodes of Wintergraze 70 and White BK-1 were compared during the cold hardening and rejuvenation periods.The results showed that during the cold hardening period,rye mainly improved its cold resistance by accumulating osmoregulatory substances in leaves and tillering nodes.During this period,the semi lethal temperature gradually decreased,and the semi lethal temperature of White BK-1 reached -9.93 ℃,which was 1.95 ℃ lower than Wintergraze 70 while the soil freezing.The increase of soluble sugar and soluble protein contents in the leaves and tillering nodes of White BK-1 was greater than those of Wintergraze 70,which meant these physiological pathways played an important role in improving the tolerance of White BK-1 to low temperatures in winter.Research on the rejuvenation period found that with the extension of low temperature stress time,the content of malondialdehyde first increased and then decreased.Rye resisted the low temperature of rejuvenation period by increasing the content of osmoregulatory substances,improving the activity of antioxidant enzymes in leaves and regulating the activity of antioxidant enzymes in tillering nodes.White BK-1 suffered less damage under the low temperature stress during rejuvenation period,and its malondialdehyde content was lower than Wintergraze 70 after recovery.In addition,after recovery,two varieties of rye accumulated higher contents of proline and soluble protein in the tillering nodes,which providing sufficient nutrients for growth and development after rejuvenation.

In order to cultivate and screen saline-alkali tolerant peanut varieties and expand the utilization area of saline-alkali soil,50 peanut varieties were used as materials,and four saline-alkali gradients,including 0,0.4%,0.8% and 1.2%,were set by potting to screen saline-alkali tolerant varieties. The results showed that under the saline-alkali stress of 0.8% in the germination stage,the seeds of 50 varieties were significantly differentiated in terms of saline-alkali tolerance,and the germination rate and seedling vigor of 8 varieties remained above 90% and 85%. The evaluation results of various physiological indexes at the seedling stage showed that the root indexes and aerial traits did not change much under low concentration (0.4%)saline-alkali stress,but showed different degrees of decline under medium (0.8%)and high (1.2%)concentration stress,and the reduction range was 22%—54% and 30%—56%,respectively. In addition,chlorophyll indexes showed an upward trend with the increase of saline-alkali concentration,with an increase range of 12%—13%. The comprehensive evaluation results of peanut using the excellent index showed that 23,5 and 0 varieties performed excellently under the saline-alkali stress of 0.4%,0.8% and 1.2%,respectively. Correlation analysis revealed that,there was no significant correlation between saline-alkali tolerance stress at the germination stage and the seedling stage,indicating that there were different mechanisms of saline-alkali tolerance during germination and seedling stages. The above results concluded that under the conditions of pot cultivation,moderate concentration (0.8%)saline-alkali was suitable for the screening of peanut saline-alkali tolerance varieties. According to the comprehensive excellence index,five saline-alkali tolerant peanut varieties were selected,namely Jihua 572,Tang 3432,Jinonghua 31,Yihua 16 and Yuhua 191,which provided a basis and germplasm resources for the promotion and cultivation of salt alkali tolerant peanut varieties.

To scientifically and accurately identify foxtail millet germplasm resources,strengthen the management of millet germplasm resources and protect new varieties.33 pairs of SSR markers were selected to establish molecular identity cards for 94 foxtail millet germplasm resources.A total of 500 SSR markers covering 9 linkage groups of foxtail millet chromosomes were screened,of which 33 markers showed polymorphic bands and stable amplification,these markers were used to establish molecular ID of 94 foxtail millet varieties.A total of 203 polymorphic bands were obtained.The number of alleles revealed by each marker was 3—10 with an average of 6.2.The polymorphism information content(PIC)values for the SSR loci ranged from 0.429 to 0.864 with an average of 0.740.Ten markers,including b185,b260,b224,b103,b225,CAAS1044,b186,b253,b105 and CAAS3008,were identified to be efficient in the variety ID construction.The IDs of 94 foxtail millet varieties were established by the 10 pairs of markers,which could clear distinguish all the varieties tested.

In order to determine the suitable sowing date of green manure for multiple cropping rape in Hetao irrigation area,field experiments were carried out in Hangjinhou Banner,Bayannur City,Inner Mongolia Autonomous Region in 2019 and Dalad Banner,Ordos City in 2022.There were five sowing dates in Hangjinhou Banner experimental site from July 26th to August 15th,2019,and one sowing date was set every five days,which were sown on July 26th,July 31st,August 5th,August 10th and August 15th,respectively.Similarly,from July 22nd to August 11th,2022,the experimental site in Dalad Banner was set up with five sowing dates,and one sowing date was set every five days to study the effects of different sowing dates on the green manure biomass,nutrient content and nutrient accumulation of rape.The results showed that although the biomass level and climate conditions of the two experimental sites were different,the overall trend showed a decline with the delay of sowing date.Compared with the first sowing stage,the rape biomass in the fifth sowing stage decreased by 90.3% and 75.4% respectively,and the average active accumulated temperature,effective accumulated temperature and sunshine hours in the two-point growth period decreased by 469.9 ℃,409.9 ℃ and 179.1 h,respectively.At the same time,when sowing in the first,second and third stages,compared with the nutrient input (N 27.3 kg/ha,P₂O₅ 34.5 kg/ha),the nutrient input of rape was higher than the input,and the amount of carbon and potassium returned to the field was at least C1 800 kg/ha and K₂O 200 kg/ha.Therefore,considering the biomass of rape green manure and the amount of nutrients returned to the field,rape could be sown early after wheat harvest in Hetao irrigation area,making full use of light and temperature resources to promote rape growth and nutrient accumulation.In order to achieve more than 5 t/ha of rape returning to the field and obtain a higher amount of nutrients returning to the field,at least 1 300 ℃ of active accumulated temperature,1 100 ℃ of effective accumulated temperature and 640 h of effective sunshine hours should be accepted during the rape growth period.

To investigate the function of ATP-binding cassette transporter subfamily G11(ABCG11)in xerophytic economic shrub Lycium barbarum ssp. Bianguo,using the Lycium barbarum ssp. Bianguo as the material. The cDNA sequence of Lycium barbarum ssp. Bianguo LbABCG11 gene was obtained by RT-PCR and RACE method, the sequence patterns were analyzed by bioinformatics software. Quantitative Real-time PCR(qRT-PCR)was used to analyze the expression patterns of LbABCG11 in different organs of Lycium barbarum ssp. bianguo under salt treatment,osmotic stress and abscisic acid(ABA)treatments.The results showed that LbABCG11 gene contained 2 971 bp in full length and a 2 130 bp open reading frame,encoding 710 amino acids.The protein contained six transmembrane regions, the calculated molecular weight, isoelectric point, adipose index, instability index and average hydropathicity was 79.39 ku,8.42, 89.63, 35.52, 0.070, the encoded protein chemical formula was C₃₅₉₀H₅₅₇₇N₉₃₅O₁₀₂₇S_35. LbABCG11 is a stable alkaline hydrophobic protein.The protein contained a nucleotide-binding domain(NBD)and a transmembrane domain(TMD),which were arranged in the form of NBD-TMD.LbABCG11 was a white-brown complex (WBC)half-size molecular transporter.The secondary structure of the protein was mainly composed of α-helix and random coil, accounting for 45.21% and 33.66%, respectively, which was consistent with the predicted results of the tertiary structure. Phylogenetic analysis showed that LbABCG11 had closer genetic relationship with ABCG11 from Solanum lycopersicum and Capsicum barbarum, but has low homology with ABCG11 from Arabidopsis thaliana and Chenopodium quinoa. LbABCG11 gene was expressed in some organs of Lycium barbarum ssp. bianguo,including leaves,stems and roots.The LbABCG11 expression level was induced by NaCl,osmotic stress and ABA,manifesting that LbABCG11 was involved in the stress response of xerophytes.

The aims of this study were to explore the genetic diversity,phylogenetic evolution and genetic relationship from six yak groups of Tibetan yaks by mtDNA ND1.To provide a theoretical basis for the genetic diversity,historical evolution,the protection and utilization of genetic resources of Tibetan yaks.The sequences of ND1 gene protein coding region(CDS)in 95 individuals of Pali yak,Jiali yak,Gongbujiangda yak,Sibu yak,Jiangda yak and Leiwuqi yak were analyzed by PCR and direct sequencing.DNAMAN,DNASP 5.1,Mega 7.0,Arlequin 3.5.2 and other software were used to analyze the sequence polymorphism,haplotype diversity,genetic distance and constructed the haplotype network map and phylogenetic tree.The result showed that the sequence length of CDS region of ND1 gene of Tibetan yaks population were 956 bp,and a total of 78 mutation loci and 16 haplotypes were detected and the average haplotype diversity(Hd)and nucleotide diversity(Pi)were 0.670 and 0.004 21,Tajima's D was all negative.According to the molecular variance analysis of population genetic differences,the degree of variation within Tibetan yaks was greater than among populations;the degree of differentiation between Sibu yak and Jiali yak was large,and the degree of differentiation among most other groups was medium or weak.In addition,cluster analysis found that Leiwuqi yak was grouped into a single group,Jiali yak,Gongbujiangda yak,Sibu yak and Jiangda yak were first grouped into one group,and then grouped with Pali yak;the 16 haplotypes were divided into two clusters,suggesting that there may be two maternal origins in Tibetan yaks.The cluster analysis with other cattle breeds showed that Bos grunniens,Bos taurus and Bos javanicus may be the mixed maternal origin of Tibetan yaks.The genetic diversity of Tibetan yaks was very rich,among which the genetic diversity of Leiwuqi yaks was the most abundant,six yak populations had two maternal origins.

In order to explore the role of RPM1 in sorghum disease resistance,a sorghum SbRPM1 gene was obtained from sorghum smut resistant variety SX44B by homologous cloning method.The bioinformatics analysis results showed that the total length of the cDNA of SbRPM1 gene was 2 802 bp,encoding 933 amino acids,and its protein had a theoretical molecular weight of 106.1 ku and an isoelectric point of 7.11,which was a hydrophilic protein.The SbRPM1 protein had no transmembrane structure,and its subcellular localization was in the cytoplasm.Conservative domain analysis showed that SbRPM1 protein contained RX-CC-like,NB-ARC and LRR domains,and belonged to CNL proteins in the NLRs family.Phylogenetic analysis showed that SbRPM1 protein was most closely related to the RPM1 protein of Miscanthus lutarioriparius.The expression pattern of SbRPM1 gene was detected by Real-time quantitative PCR,and the results showed that the expression of SbRPM1 gene was higher in leaves and inflorescence,followed by roots,and the lowest in stem.The expression of SbRPM1 gene was significantly up-regulated at 24—72 h in disease-resistant varieties after inoculation with Sporisorium reilianum pathogen,suggesting that this gene could be induced by S.reilianum and played an important role in sorghum disease resistance.In this study, the CDS sequence of the SbRPM1 gene was cloned for the first time in sorghum, and the structure, nature and expression of the gene were characterized.

Luchuan pig myoblast determining gene 1(MyoD1)was cloned,bioinformatics analysis was performed,and its expression was analyzed in different tissues of Luchuan pigs,to explore the role of MyoD1 gene in the growth and development of skeletal muscle.Published MyoD1 gene of wild boar sequence on NCBI as the template,designed specific primer,cloned the CDS region of MyoD1,and compared the similarity with wild boar,cattle,sheep,human,mouse and rat gene sequence,constructed system tree for bioinformatics analysis by online software.Then,the relative expression of MyoD1 gene in different tissues of Luchuan pigs by RT-PCR.The CDS of MyoD1 gene in Luchuan pig was 960 bp,encoding 319 amino acids,five base mutations.The similarities of MyoD1 genes between Luchuan pig and wild boar,cattle,sheep,human,mouse and rats were 99.2%,93.0%,92.3%,90.0%,84.3% and 84.0%,respectively.Total number of MyoD1 gene atoms in Luchuan pigs was 4 680,molecular mass of the protein was 33.99 ku,molecular formula was C₁₄₅₇H₂₂₉₆N₄₄₂O₄₇₁S₁₄;the instability coefficient was 63.87,indicating a lack of stability;isoelectric point was 5.63,which belongs to the acidic protein;no transmembrane structure existed,found 35 phosphorylation sites and 1 glycosylation site;protein secondary structure was dominated by an irregular coil,which accounting for 60.69%.The MyoD1 gene in Luchuan pigs had the highest expression in the longest dorsal muscle,which was significantly higher than in other tissues,while kidney had the lowest expression.MyoD1 gene was expressed in all tissues of Luchuan pigs and mainly in the longest dorsal muscle,we speculated that MyoD1 gene might play a crucial role in the muscle growth and development of Luchuan pigs.

Silk color is an important agronomic trait for determining distinctiveness and uniformity of maize variety.In order to analyze the genetic mechanism of silk coloring characteristics of anthocyanins in maize,a doubled haploid(DH)population with 213 lines derived from green silk inbred line WL134 and purple silk inbred line D7 was used for QTL mapping analysis under the environment of 2022 and 2023,respectively.The results showed that there were significant differences in silk coloring characteristics of anthocyanins among different lines and years,the heritability was 0.864.A total of 9 QTLs were detected in two years.These QTLs with phenotypic variation explained(PVE)ranging from 4.83% to 9.26% were detected on chromosomes 2,3,5,6,8,and 9 of maize.A stable and repeatable site qSC5 located between 19.15 Mb and 19.80 Mb was detected on chromosome 5 in two-year data.The LOD scores of the major QTLs were 4.65 and 5.76 in 2022 and 2023 respectively,with phenotypic variation explained(PVE)of 7.22% and 7.17%.It was a new site for regulating silk coloring characteristics of anthocyanins compared with previous studies in maize.Based on SNP markers on both sides of qSC5,90.91% of the genotypes in extreme purple silk DH lines were CCCC,while only 44.00% of the genotypes in green silk DH lines.This marker was significantly correlated with silk color in maize and might link to key genes regulating silk color characteristics of anthocyanins.

In order to control the amount of organic manure in the greenhouse vegetable management system,reduce the load of nitrogen and phosphorus in the greenhouse soil and reduce the risk of non-point source pollution of nitrogen and phosphorus in the greenhouse vegetable,a 3-year field experiment,was conducted,and a total of 5 treatments were set.They were the farmers' traditional amount of organic manure in this study area(FP,i.e.the control treatment),the amount of 50% of farmers' traditional amount of organic manure(FP-M,i.e organic manure reduction treatment),and catch crop planted in the fallow period on the basis of the FP-M treatment with forage maize,forage sorghum and legumes incorporation(FP-M+C,FP-M+S and FP-M+L,respectively).In FP-M+C,FP-M+S and FP-M+L treatment,the straw was returned in situ before vegetable transplanting.The leaching liquor was collected in situ,and nitrogen and phosphorus leaching amount,soil organic matter, {{\mathrm{NO}}_3}^{-}-N,Olsen-P and vegetable yield were measured,to study the effect of substitution of catch crops for organic manure during the fallow period.The results showed that,there was no difference in vegetable yield between FP treatment and FP-M,FP-M+C,FP-M+S and FP-M+L treatment.However,FP-M+C and FP-M+S treatments had significantly higher organic matter content in 0—40 cm soil compare to FP treatment.Compared with FP treatment,the leaching amount of total nitrogen in the FP-M,FP-M+C,FP-M+S and FP-M+L treatment showed a significant decrease in the fallow period,and decreased by as much as 13.9%,59.5%,65.4% and 54.5%,respectively;in the vegetable growing period,and decreased by as much as 38.3%,48.5%,39.0% and 24.0%,respectively.Compared with FP treatment,the leaching amount of total phosphorus in the FP-M,FP-M+C,FP-M+S and FP-M+L treatment showed a significant decrease in the fallow period,and decreased by as much as 42.3%,53.9%,45.4% and 49.1%,respectively;in the vegetable growing period,and decreased by as much as 37.7%,33.8%,27.7% and 26.3%,respectively.The {{\mathrm{NO}}_3}^{-}-N and Olsen-P contents soil profile of the FP-M,FP-M+C,FP-M+S and FP-M+L treatments were significantly decreased compared with FP treatment,in 80—200 cm and 20—80 cm,respectively.Therefore,that reducing 50% of the organic manure by farmers' practice and planting catch crops in the fallow period and returning the straw in situ before vegetable transplanting can increase organic matter content in 0—40 cm soil in the study area.Obviously,the organic manure can be substituted with catch crops.It reduces the risk of nitrogen and phosphorus pollution in greenhouse vegetable while decreasing nitrogen and phosphorus leaching.

In order to clarify the sequence characteristics of the MYB62 transcription factor,expression changes after stress,and further explore the biological function of the MYB62 transcription factor,kohlrabis were used as plant materials.MYB62 transcription factors were obtained by homologous cloning method and bioinformatics was analyzed.Spatiotemporal expression analysis of MYB62 and its expression after stress were performed by Real-time PCR.Results of gene cloning showed that gDNA length of BocMYB62 gene was 1 353 bp,the length of CDS was 837 bp,which contained four exons and three introns and encoded 278 amino acids.The sequence structure analysis showed that BocMYB62 was a hydrophilic protein with two SANT-MYB domains,belonging to the R2R3-MYB type in the MYB transcription factor family.The spatial structure prediction revealed a typical α-helix structure.Phylogenetic analysis indicated that BocMYB62 was closely related to MYB62 in Brassica napus.The spatiotemporal expression results showed that BocMYB62 was consistently higher in green kohlrabi than that in purple with clear tissue specificity.The expression of BocMYB62 was significantly increased during drought stress with the highest expression under 12 h.The BocMYB62 expression was significantly lower than control and the lowest at 4 ℃ cold stress.These results inferred that BocMYB62 may be involved in the regulation of anthocyanin biosynthesis and the regulation of stress.It provides a theoretical basis for the subsequent functional identification of MYB62 transcription factors.

The full-length sequence of Diapausin,a novel antimicrobial peptide gene,was obtained from the third instar of oriental armyworm(Mythimna separata)by RT-PCR and RACE.And then,the amino acid sequence of Diapausin gene was analyzed by bioinformatics software.Finally,the prokaryotic expression system was used to induce the protein expression.This study would lay a theoretical foundation for further parsing of the function and structure of Diapausin gene in M.separata. Results showed that the novel antimicrobial peptide gene Diapausin was successfully cloned from M.separata and named Mysdiap (GenBank No.AZJ51075).The total length of this gene was 537 bp,with the 5' and 3' non-coding ends of 63,279 bp,respectively.The full length of the open reading frame was 195 bp,encoding 64 amino acid residues.The molecular weight and the isoelectric point of the protein were 7.13 ku and 5.59,respectively.The first 24 N-terminal amino acid residues of Mysdiap were signal peptide sequences.Multiple alignment of amino acid sequences showed that Mysdiap had a highly conserved region ECCRAHG.The recombinant expression plasmid pET-Mysdiap was successfully constructed and induced to express proteins by IPTG in Escherichia coli.The molecular weight of the expressed recombinant protein was about 20 ku,which could exist in both inclusion bodies and soluble proteins.In summary,the full-length sequence of the novel antimicrobial peptide gene Mysdiap was cloned from M.separata and successfully induced to express proteins in E.coli.

Modern soybean cultivars typically display yellow to brown pods,while their wild ancestral specie,Glycine soja,possesses black pods.Pod color is an important domestication trait and phenotypic characteristic,which is strongly related to pod blasting habit and avoidance of predation.Two alleles were certified to control the pod color in soybean,among which the brown pod L2 gene has not been identified.In order to identify L2 gene on the soybean genome,and provide a theoretical basis for functional analysis and breeding application of brown pod related genes in soybean.The cultivated varieties Zhonglongyou 203(yellow pod)and wild varieties FF1235(black pod)were used as parents to generate an F₂ segregating population for genetic analysis in this study.The BSA-seq was performed using two gene pools which were constructed by brown pod and yellow pod individuals from the F₂ population,respectively.On this basis,recombinant exchange individuals were analysed.The results showed that brown pod was a quality trait controlled by a pair of alleles in soybean.The brown pod L2 gene was located in the 0—0.75 Mb region of Chromosome 3.By further use of 7 polymorphic InDel markers in fine mapping,the candidate interval was finally delimited between Indel-L2-3 and Indel-L2-6 with 344 kb physical distance.There were 32 candidate genes in the interval,among which Glyma.03G005700 gene was annotated as isopropylmalate polymerase.Glyma.03G005700 gene is highly homologous to the discovered black pod gene L1 (Glyma.19G120400),which may be responsible for converting 4-hydroxypyruvate into eucomic acid and piscidic acid,and may be a key gene in the regulating the formation of brown pod in soybean.

In order to determine the effect of allele variation in DEP1 locus on quality traits and metabolic profile of grains in rice,the high quality japonica cultivar Daohuaxiang and its erect panicle type mutant dep1-1 were used.The grain quality traits and differential metabolites between wild-type and mutant dep1-1 were measured.The results showed that the mutant dep1-1 had a single base A inserted into the fifth exon of the DEP1 gene,leading to premature termination of translation and the deletion of the cysteine-rich region in G-protein gamma subunit.Compared with wild-type cultivar,the grain length,appearance,viscosity,balance,taste value,peak value,recovery value and peak time of mutant dep1-1 were significantly decreased,while amylose content, acidity value and hardness were significantly increased.Compared with wild-type cultivar,37 metabolites were significantly up-regulated in dep1-1,while 45 were significantly down-regulated.KEGG analysis showed that these differential metabolites were mainly enriched in galactose metabolism,fatty acid biosynthesis,starch sucrose metabolism,linoleic acid metabolism,ABC transporters,and phenylpropanoid biosynthesis pathways.Compared with wild-type cultivar,the contents of sucrose,D-raffinose,raffinose,D-(+)-maltose D-(+),betaine and indole of grains significantly increased in dep1-1,while those of stearic acid,palmitic acid,L-leucine and indole-3-acetic acid significantly decreased.These differential metabolites preliminarily explained the physiological and biochemical mechanism of grain quality changes in dep1-1 mutant.

The aim of this study was to obtain the serum inflammatory factor content,spleen pathological changes and transcriptome expression profile characteristics of diarrheic piglets with Clostridium perfringens type C,and to provide a reference for the diarrhea prevention and treatment.Six 7-day-old binary piglets with good health and similar body weight were randomly selected and divided into two groups,namely control group (CS) and treatment group (TS).Each piglet in the TS group was fed with 1 mL bacterial solution (1×10⁹ cfu/mL) every day for 5 consecutive days.Then,piglets in the CS group were fed with culture medium without the bacterial solution.Blood samples were collected and serum was separated on 0,1,3 and 5 days after infection.The serum IL-8,IFN-α and IFN-γ concentration were detected by ELISA.After the experiment,piglets were slaughtered to collect the spleen.The partial spleen was collected for HE staining to observe the pathological changes.Additionally,differentially expressed (DE) genes in the spleen were obtained using RNA-Seq.GO terms and KEGG signaling pathways enrichment analysis were performed.The key spleen genes during the infection of piglets with C.perfringens type C were verified by RT-qPCR.The spleen of piglets in the TS group displayed obvious inflammation,neutrophil infiltration,accompanied by degeneration and necrosis of parenchymal cells;on the 3rd and 5th day,the levels of IL-8,IFN-α,and IFN-γ in the CS group serum were extremely significantly lower than those in the TS group (P<0.01);a total of 1 079 DG genes was screened between TS group and CS group,including 583 up-regulated genes and 496 down-regulated genes.DG genes were mainly enriched in GO terms such as cell response to interferon-β,cell response to DNA damage stimulation,and negative regulation of inflammatory response.Differentially expressed gene were mainly enriched in KEGG signaling pathways such as tuberculosis,hepatitis B,and B cell receptors.These GO terms and KEGG signaling pathways were involved in piglet resistance to C.perfringens type C infection;the RNA-Seq results of the selected candidate genes (such as JAK3,TLR3,ILF2,CXXC1) resistant to C.perfringens type C infection were consistent with the RT-qPCR results.The above results indicated that JAK3,TLR3 and other genes played important roles in the process of piglet spleen resistance to C.perfringens type C infection through key signaling pathways.

HD-Zip transcription factors play an important regulatory role in stress resistance in plants,in order to reveal the characteristics and potential functions of the HD-Zip gene family in the hemp genome,and to provide important clues for further study on the regulation of CsHDZ gene under drought stress.Here this study based on the reference sequence in the hemp whole genome database,perform a genome-wide analysis of HD-Zip gene family by bioinformatics methods in hemp,and their physical and chemical characteristics,evolutionary relationship,gene structure,the cis-acting elements and transcript patterns in drought stress were analyzed.The results showed that a total of 20 hemp HD-Zips (CsHDZs),they were non-evenly distributed on seven chromosomes,and classified into four distinctive subfamilies.Their amino acid sequence size and isoelectric point were 204—837 aa and 4.54—9.04,they were all unstable proteins,and subcellular localization analysis indicated that HD-Zip protein was targeted to the nucleus.Analysis of protein conserved motifs showed that there was a total of 10 conserved motifs in CsHDZs,of which motif 1 and motif 9 were shared by all numbers.In addition,the regulatory element analysis showed that a variety of elements related to abiotic stresses were identified in the promoter region of CdHDZs.The results of Real-time Fluorescence Quantitative analysis showed that the transcription level of CsHDZ1/8/10 gene had no significant difference in the early stage of drought,but was significantly upregulated in the late stage.The results indicated that CsHDZs gene was involved in the stress response process and could actively respond to drought.

To investigate the effect of exogenous melatonin (MT) on salt tolerance of soybean seedlings,and to screen the appropriate application concentration under different salt stress.The soybean variety Tianyou-2986 was used as the test material,and 3 salt concentrations (low salt S₃:3 g/L,medium salt S₅:5 g/L,high salt S₇:7 g/L ) and 6 MT concentrations were set(M₀:0 μmol/L,M₁:25 μmol/L,M₂:50 μmol/L,M₃:75 μmol/L,M₄:100 μmol/L,M₅:150 μmol/L),the morphological parameters,biomass,antioxidant enzyme activity and osmoregulatory substance content of soybean seedlings were analyzed.With the increase of salt stress,the root morphological parameters,biomass,root-shoot ratio,antioxidant enzyme activity and osmoregulatory substance content of soybean seedlings decreased,while the malondialdehyde content increased.Under low salt (S₃) and medium salt (S₅) stress,The number of total length,lateral roots,SOD and POD of 50 μmol/L MT were increased by 52.30%,19.98%,74.10%,40.03% (low salt) and 68.52%,19.24%,81.72% and 37.42% (medium salt),respectively.Under high stress (S₇),75 μmol/L MT increased by 71.17%,19.11%,80.79% and 27.01%,respectively.Under salt stress,exogenous 25—100 μmol/L MT promoted soybean seedling growth and improved salt tolerance to different degrees.The overall evaluation of the affiliation function showed that 50 μmol/L MT was the most effective in alleviating salt damage under low and medium salt stress,and the suitable concentration of MT was 75 μmol/L under high salt stress,and the main reason for the alleviation of salt damage in soybean seedlings was that MT increased the activities of antioxidant enzymes and osmoregulatory substance content,and reduced the content of malondialdehyde,which could alleviate oxidative and osmotic stresses of soybeans under salt stress.

In order to study the effect of applying N,P and K fertilizer on the yield and nutrient use efficiency of oil sunflower under the double cropping pattern of winter wheat-summer oil sunflower in Southern Shanxi,a field plot fertilizer experiment was carried out with Linkui 4 from 2019 to 2021.Four fertilization treatments including NPK,PK,NK and NP were set up.The effects of different fertilization treatments on yield response,agronomic efficiency and fertilizer use efficiency of N,P,K were analyzed.The results showed that the three-year average yield of oil sunflower treated with NPK was 3 634 kg/ha(3 502—3 818 kg/ha),which was 19.6%,12.1%,13.7% higher than that of PK/NK/NP,respectively.PK treatment had the lowest yield.The average yield responses of N,P and K fertilizers were 588.8(481.0—704.7 kg/ha),391.8(296.0—488.7 kg/ha),438.2 kg/ha(329.3—629.7 kg/ha),respectively.The agronomic efficiencies of N,P and K were 3.8(2.8—4.9 kg/kg),3.1(2.1—4.1 kg/kg),2.9 kg/kg(2.2—4.2 kg/kg),respectively.The aboveground accumulations of N,P and K were 174.4(172.5—176.8 kg/ha),60.0(58.3—61.8 kg/ha),241.1 kg/ha(236.2—246.8 kg/ha),respectively.The nutrient use efficiency of N,P,K was 36.3%(36.0%—36.7%),15.7%(15.3%—16.2%)and 47.1%(46.0%—47.9%),respectively,of which K>N>P.So the application of N,P and K fertilizer could not only significantly increase the oil sunflower yield but also the aboveground accumulation of N,P and K.Among the three kinds of NPK fertilizers,N increased the yield most,and K uptake by the aboveground part and utilization rate of oil sunflower was the highest.Therefore,in the production we should pay attention to the reasonable application of N,P and K,while N and K should be supplemented by seed fertilizer and topdressing.

Calcium and potassium are important mineral nutrient elements in wheat.It is significant to explore the related genetic mechanisms and effects on human nutritional health.To provide a theoretical basis for biofortification breeding of trace elements in wheat grains,we used 164 F₆ recombinant inbred lines(RILs)derived from Avocet/Chilero(AC)and 175 F₆ RILs derived from Avocet/Huites(AH).Our investigation focused on phenotypic variations in grain calcium(GCa)and grain potassium(GK)content in five environments.QTL mapping was conducted with diversity arrays technology(DArT)chip.Nineteen QTLs associated with grain calcium content were identified,distributed on chromosomes 1A,1D,2A,2B,3A,3D,4A,4B,4D,5A,5B,7A,7B,and 7D,explaining 3.23%—16.29% of phenotypic variation.Simultaneously,23 QTLs linked to grain potassium content were identified on chromosomes 1B,2A,2B,3A,3B,4A,4D,5A,6A,6B,and 7D,explaining 3.31%—24.66% of phenotypic variation.QGCa.haust-1A,QGCa.haust-AC-5A and QGK.haust-AC-2A.2 were located in multiple environments.QGCa.haust-1A and QGCa.haust-AC-5A explained 7.82%—12.72% and 9.68%—15.57% of phenotypic variation,and the physical intervals were 498.67—532.21 Mb and 461.52—486.26 Mb,respectively.QGK.haust-AC-2A.2 explained 8.15%—15.20% of phenotypic variation,with a physical range of 354.61—462.37 Mb.The genetic effect analysis of QGCa.haust-1A,QGCa.haust-AC-5A,and QGK.haust-AC-2A.2 showed that each locus effectively increased the calcium and potassium content in wheat grain.Aggregation effect analysis indicated that the lines with QGCa.haust-1A and QGCa.haust-AC-5A effect loci had highly significantly higher calcium content than those with only a single locus.In summary,three stable loci of grain calcium and potassium content are mapped on chromosomes 1A,2A,and 5A,which could significantly increase calcium and potassium content in wheat grain.

In order to reveal the molecular mechanism of the response to exogenous selenium of sugar beet,seeds of the sugar beet line HD802 were used as test materials for hydroponic experiments.The concentrations of nano-selenium were set to 0(water spray,control),20,50,80,100,150,200 mg/L,respectively.When the plants grew to 8 true leaves stage,nano-selenium solution was evenly sprayed.After 24 h,the activities of superoxide dismutase(SOD),peroxidase(POD),catalase(CAT)and the content of malondialdehyde(MDA)were measured.After data analysis,50,150 mg/L nano-selenium treated leaves were selected for examination.RNA-Seq was performed to analyze the differentially expressed genes and significant enrichment pathways.The results showed that 50 mg/L nano-selenium treatment had promoting effects on the growth of sugar beet leaves,while 150 mg/L treatment had destructive effects on sugar beet leaves.A total of 9 161 DEGs were identified,of which 3 717 were up-regulated and 5 444 were down-regulated.GO functional enrichment was mainly enriched in processes such as signal transduction,cell communication, integral component of membrane, intrinsic component of membrane, primary metabolic process, organic substance metabolic process, and biological process. The metabolic pathways of KEGG mainly included plant pathogen interaction,plant hormone signal transduction,and other pathways.Transcription factor analysis involved 11 families,including AP2,zf-Dof,HLH,WRKY,HSF_DNA-bind,NAM,zf-BED and Homeobox.It found the optimal concentration for selenium treatment to promote the growth and development of sugar beet seedlings,and preliminarily screened the relevant genes responsive to exogenous selenium in sugar beet.

This study aims to explore the effects of different concentrations of zearalenone (ZEN) on the growth of bovine mammary alveolar cell-T (MAC-T) and the expression of genes related to milk fat synthesis.Firstly,MAC-T cells were treated with different doses of ZEN for 36 hours,and the number of cells was counted using a hemocytometer.Then,cells were stained and analyzed for apoptosis and necrosis to determine the appropriate dose of ZEN.Next,acommercial detection kit was used to examine the impact of ZEN on reactive oxygen species (ROS) and mitochondria number in MAC-T cells.Finally,using Real-time Quantitative PCR (RT-qPCR) technology,the influence of ZEN on proliferation,apoptosis,oxidative stress,and milk fat synthesis-related gene expression in MAC-T cells was analyzed.The results showed that low doses of ZEN (0.01—1.00 μmol/L) tended to promote MAC-T growth,whereas high doses of ZEN (5.00—10.00 μmol/L) significantly reduced the number of cells,0.10 μmol/L ZEN had no obvious effect on ROS and mitochondria number in MAC-T,but 10.00 μmol/L ZEN notably elevated ROS levels and decreased mitochondria number.RT-qPCR results indicated that 0.10 μmol/L ZEN significantly promoted the expression of proliferation genes (CDK1,CCND2),antioxidant genes (DHODH,GPX4),and anti-apoptotic genes (BCL-2),but also increased the expression of apoptotic genes (CAS-3,BAX).Whereas,10.00 μmol/L ZEN significantly inhibited the expression of proliferation genes (PCNA,CDK1,CCND2),antioxidant genes (DHODH,GPX4,AIFM2),and anti-apoptotic genes (BCL-2),while significantly promoting the expression of apoptotic genes (CAS-3,BAX).Notably,0.10 μmol/L ZEN obviously promoted the expression of milk fat synthesis-related genes (PPARγ,FASN,JAK-2),but 10.00 μmol/L ZEN significantly suppressed these genes.The aforementioned results suggested different effects of ZEN concentrations on MAC-T cells:0.10 μmol/L ZEN could promote MAC-T cell growth and milk fat synthesis-related gene expression while also inducing the expression of apoptotic genes.In contrast,10.00 μmol/L ZEN induces oxidative stress in MAC-T cells,reduces mitochondria number,inhibits the expression of genes associated with proliferation,antioxidant properties,anti-apoptotic properties,and milk fat synthesis,while promoting the expression of apoptotic genes,leading to cell apoptosis.