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

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.

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

In order to uncover key genes involved in flax's response to drought stress and elucidate the underlying molecular mechanisms,thereby providing theoretical support for breeding drought-resistant varieties.Using roots of the drought-resistant flax cultivar Tianya 13 as experimental material,drought stress was simulated with different concentrations(5%,10%,20%,30%)of polyethylene glycol(PEG).Based on the analysis of physiological indicators such as root length,root thickness,and dry/fresh weight ratio,20% PEG was identified as the optimal stress concentration,under which the above indicators were significantly reduced by 31.63%,30.50%,and 13.39 percentage points,respectively,compared to the control.Subsequently,Illumina transcriptome sequencing was performed on roots subjected to drought stress at different time points(8,16,24 h),identifying 5 428,5 304,and 3 603 differentially expressed genes(DEGs)at each time point,respectively.KEGG enrichment analysis revealed that these DEGs were significantly enriched in key pathways such as carbohydrate metabolism(12 genes),betaine biosynthesis(4 genes),and the antioxidant defense system(36 genes).qRT-PCR validation of selected DEGs confirmed expression trends consistent with the RNA-seq results,verifying data reliability.This study systematically reveals,that flax enhances drought tolerance through the coordinated regulation of osmotic adjustment substance(e.g.,betaine)synthesis,energy metabolism reprogramming(carbohydrate metabolism),and reactive oxygen species scavenging mechanisms.It provides important theoretical foundations and candidate gene resources for molecular design breeding in flax.

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.

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

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

In order to understand the function of SibHLH19 in foxtail millet,the CDS sequence and promoter sequence of SibHLH19 gene were separately cloned with the leaf cDNA and genomic DNA from resistance material Shilixiang as template by PCR.Promoter cis-acting elements and biological characteristics were analyzed using bioinformatics online tools.Then the expression patterns of SibHLH19 in different tissues and during the process to rust resistance were surveyed by qRT-PCR,respectively.Lastly the prokaryotic expression characteristics for the gene were detected by SDS-PAGE,laying a theoretical foundation for further research on SibHLH19 gene function and disease resistance mechanism.The results showed that the CDS sequence of the SibHLH19 transcription factor was 843 bp in length,encoding a total of 280 amino acids,the predicted protein molecular weight was 29.97 ku.The theoretical isoelectric point was 5.85,and the encoded protein chemical formula was C₁₂₉₆H₂₀₇₁N₃₉₇O₄₀₀S₁₁,containing a bHLH conserved domain,belonging unstable hydrophilic protein.The largest element of the protein's secondary structure was random coils,and the smallest element was a β-turn.Evolutionary analysis showed that SibHLH19 had the higher homology to the amino acid sequences of Panicum miliaceum (RLM85279.1),Panicum hallii (PUZ71581.1)and Panicum virgatum (XP_039835205.1),and had the lowest homology with Triticum aestivum(KAF7059972.1)and Aegilops tauschii subsp.strangulata (XP_040244423.1).The analysis of the promoter cis-acting elements showed that there were multiple response elements such as hormones and stresses in the promoter region of the SibHLH19 gene.Tissue expression analysis showed that the gene was mainly expressed at the seedling stage with the highest expression in the aboveground part,and was almost no expression at the booting stage.Within 24 hours of the response to the biotic stress of rust disease in foxtail millet,the SibHLH19 gene expression was up-regulated at 8 and 16 h in the disease resistance response,while its expression was only slightly up-regulated at 16 h and down-regulated at the rest of the time points in the susceptible response.It was speculated that SibHLH19 played a positive regulatory role in the resistance response to rust disease in foxtail millet.The constructed prokaryotic expression vector pET30a-SibHLH19 could express the SibHLH19 fusion protein with an apparent molecular weight of about 44 ku after being induced by 0.1 mmol/L IPTG.

The SiPRR73 gene was cloned from Yangu 11 using RT-PCR technology,and through analyzing tissue-specific expression,responsive features of SiPRR73 to different photoperiods,photo-thermal combinational treatments and five abiotic stress treatments,the regulation mode of photoperiod and temperature on SiPRR73,and the responsive pattern of SiPRR73 to abiotic stresses in foxtail millet were explored. The results showed that totally 2 928 bp cDNA sequence of SiPRR73 was obtained from Yangu 11,which included 2 283 bp CDS region,encoding 760 amino acids. The SiPRR73 proteins of C4 crops including Panicum miliaceum,Panicum hallii,Sorghum bicolor and Zea mays showed relatively close relationship with SiPRR73. The second parietal leaf was the highest expression tissue of SiPRR73,but the expression level at root,stem and panicle tissues was relatively lower. The expression level of SiPRR73 was higher at light period than that at dark period under both short-day and long-day conditions,and during the whole vegetative growth phase,SiPRR73 showed higher expression level under long-day compared to short-day,which indicated that the expression of SiPRR73 was induced by light and controlled by photoperiod. The temperature determined expression peak number of SiPRR73 and the photoperiod determined occurrence time of expression peaks,so temperature and photoperiod participated in regulating of SiPRR73 expression mutually. PEG and low temperature stresses induced SiPRR73 expression totally,NaCl induced SiPRR73 expression at early stress stage,but inhibited it at later stress stage. Fe stress inhibited SiPRR73 expression at early stage,but induced it at later stage. ABA stress caused the close responsive feature of SiPRR73 to NaCl. This study indicated that SiPRR73 showed light-dependent expression feature,and photoperiod and temperature regulated SiPRR73 by interaction pattern,suggesting that SiPRR73 participated in adaptability regulation process to different photo-thermal conditions and might play a certain role in coping with drought,low temperature,ABA,NaCl and Fe stresses in foxtail millet.

To investigate the expression characteristics of OsASL1.1 and its role in the drought stress response,the subcellular localization of OsASL1.1 protein and cis-acting elements in the promoter region of OsASL1.1 gene were analyzed.Additionally,Real-time Quantitative PCR(qRT-PCR)was used to examine the expression patterns of OsASL1.1 in response to abscisic acid(ABA),mannitol,and polyethylene glycol 6000(PEG6000).CRISPR/Cas9 gene editing technology was employed to generate Cas9-OsASL1.1 mutants with Kitaake as background,and the mutation types were characterized.The abscisic acid(ABA)sensitivity and drought tolerance of mutants were analyzed.The results indicated that OsASL1.1 protein was primarily localized in plastids.The promoter region of OsASL1.1 contained cis-acting elements responsive to low temperatures and plant hormones(ABA and auxin),including five ABA responsive cis-acting elements(ABREs).OsASL1.1 expression was induced by ABA,mannitol and PEG6000.Two homozygous mutant types,Cas9-6 and Cas9-12,were generated.Cas9-6 harbored a 17 bp insertion that caused premature translation termination,whereas Cas9-12 had a 12 bp deletion that resulted in the loss of four amino acids.Under ABA treatment,root elongation of Cas9-OsASL1.1 mutants and wild type were inhibited;however,the inhibition was less severe in the mutants than that in the wild type.Under 150 mmol/L mannitol treatment,the survival rates of both Cas9-6 and Cas9-12 mutants were significantly lower than that of wild type,with Cas9-6 showing the lowest survival rate.In summary,the Cas9-OsASL1.1 mutants show reduced sensitivity to ABA and decreased drought tolerance,indicating that the loss of OsASL1.1 function diminishes rice sensitivity to ABA and drought tolerance.

This study aimed to elucidate the compositional differences and molecular regulatory mechanisms of anthocyanins in kernels of fresh-eating maize with different colors by integrating metabolomic and transcriptomic analyses,thereby providing candidate genes and a theoretical basis for molecular breeding of colored fresh-eating maize.White WN2000,yellow HN,colored CN1,and black HTN188 were used as materials.Metabolomics was employed to identify anthocyanin-related metabolites and screen differential metabolites;transcriptome sequencing was conducted to obtain differentially expressed genes(DEGs).Metabolite gene associations were analyzed through GO annotation and KEGG pathway enrichment,and key structural genes and transcriptional regulators were mined within the anthocyanin biosynthetic pathway.The results showed that 49 anthocyanin-related metabolites were identified in total.Major anthocyanins,including cyanidin,peonidin,and pelargonidin,accumulated significantly in HTN188,indicating stronger pigment deposition.Transcriptome analysis identified 12 557 DEGs,among which structural genes such as F3H,DFR,and UFGT,as well as transcription factors including MYB and bHLH,were significantly up-regulated in varieties with high anthocyanin content.Correlation analysis revealed that the expression level of Zm00001d011438 was significantly and positively correlated with total anthocyanin content,suggesting that it may play a key role in kernel anthocyanin accumulation.Overall,the integrated metabolomic and transcriptomic analyses characterized differences in anthocyanin composition and related gene expression among differently colored fresh-eating maize kernels,and identified structural genes,transcription factors,and the candidate Zm00001d011438 associated with the anthocyanin biosynthetic pathway,providing references for subsequent functional validation and genetic improvement.

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

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.

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

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

In order to further analyze the genetic basis regulating the protein content of super sweet corn stalk,this study used the BLINK model to locate the major SNP loci and predict candidate genes controlling the trait,based on the maize 56K gene chip.The results showed that during the harvest period of super sweet corn,the protein content of stalk ranged from 12.63 to 30.29 g/kg,with an average of 20.76 g/kg,showing a normal distribution.Six significant SNP loci regulating the protein content of super sweet corn stalk were identified by GWAS.These loci were distributed on chromosomes 1,4 and 6,and their contribution rates ranged from 10.07% to 18.17%.Among them,Affx-115330268 on chromosome 6 had the highest contribution rate.A total of 184 candidate genes were detected in the 200 kb confidence interval between the upstream and downstream of the significant association sites,of which 34 were annotated genes,accounting for 18.48%.Through GO functional annotation and KEGG pathway enrichment analysis,candidate genes were mainly involved in biological processes such as biological regulation,cellular processes,and catalytic activity,and were enriched in steroid biosynthesis,tryptophan metabolism,photosynthesis-antenna protein and other pathways.Based on the literature search and comparison,it was found that six key genes (GRMZM2G141473, GRMZM2G007854, GRMZM2G111319, GRMZM2G121024, GRMZM2G010353, GRMZM2G049547) that regulate the protein content of super sweet corn stalk,which are mainly involved in protein and enzyme synthesis.

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

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

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

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

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.

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

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

In order to study the tolerance of heterologously overexpressed Atvip1 gene in sorghum to defense saline-alkali stress and the corresponding growth, NaHCO₃:Na₂CO₃ of 5:1 solution with 75 mmol/L and pH 9.63 was used in sorghum at the stage of three leaves and one heart. The root growth index, chlorophyll content, antioxidant enzyme activity and MDA content were measured at 0, 4, 12, 24, 72, and 120 h of stress. The results indicated that the heterologous overexpression of Atvip1 gene could alleviate the damage of saline-alkali stress on the growth of sorghum seedlings, increase the root surface area and root volume, the number of root tips and branches, and also cause the browning of sorghum main roots to appear later and mild symphonys, and the earlier and more lateral roots occurrence. The new leaves could still be normally extended at 72 h and present little effect on the growth of aboveground. Overexpression of Atvip1 gene could increase the activity of O₂^-· resistance, decrease the content of MDA and enhance the activities of antioxidant enzymes in transgenic sorghum roots. SOD, CAT and GR had obvious effects at 4-12 h during the early stage of stress, respectively. All enzymes played roles during the middle of stress at 24-72 h. CAT and GSH-PX played important roles at the later stage of stress at 120 h. On the base of differential transcriptome analysis of saline-alkali stress, COG analysis of differentially expressed genes(DEGs) showed that defense mechanisms accounted for a relatively large proportion during various periods, and 42 DEGs related to antioxidant enzymes were obtained. Heterologous overexpression of Atvip1 gene can improve the resistance of transgenic sorghum to saline-alkali stress by alleviating the effects on photosynthesis, growth and development, reducing the damages of reactive oxygen species and membrane damage.

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

Temperature stress is one of the main nonbiological stresses that affect the quality and yield of spinach.Investigating the molecular response mechanism of spinach to temperature stress is crucial for spinach stress tolerance breeding.To provide a theoretical basis for the research of the mechanism of spinach resistant to cold stress and heat stress,this study used the cold-tolerant inbred line D3 and the heat-tolerant inbred line M10 of spinach as experimental materials and analyzed their transcriptomes and metabolomes under cold and heat stress to explore the transcriptional and metabolic mechanisms underlying spinach tolerance.Transcriptomic analysis showed that the pathways in which the DEGs were the most enriched in D3 and M10 were essentially the same under cold stress and heat stress.Metabolomics analysis showed that under cold stress,they were coenriched in the pyrimidine metabolism and lysine degradation in KEGG pathways.Under heat stress,these were mainly enriched in the tryptophan metabolism,toluene degradation,biosynthesis of various other secondary metabolites,and glycine,serine and threonine metabolism in KEGG pathways.The joint transcriptomic and metabolomic analysis indicated that through data analysis and gene annotation.SpADH(sov2g036390),SpSHMT(sov1g001130)and SpALDH-1(sov4g007150)were identified as the candidate genes for cold stress tolerance in spinach.SpALDH-1(sov4g007150),SpALDH-2(sov1g043320)and SpNPC(sov1g040610)were identified as candidate genes for heat stress in spinach.Among them,SpALDH-1(sov4g007150),which may be a regulatory gene for spinach stress tolerance,was significantly expressed under both cold and heat stress.

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.

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

In the early stage, the dwarf dwarf-12 was successfully selected by distant hybridization of dwarf sorghum and local maize.After previous studies, the dwarf gene may be controlled by br2.Since there were no adverse traits, in order to utilize and discover excellent dwarf inbred lines, and then crossed dwarf-12 with local white maize, and selected the excellent dwarf inbred line d8227. The dwarf maize material d8227 obtained by predecessors was combined with maize of different heights, and it was identified that it has good combining ability. In order to increase the germplasm resources of dwarf maize and improve the yield of maize, in-depth research was carried out.d8227 and dwarf-12 were used as research materials to compare the differences in main agronomic traits between dwarf parents and progeny to observe the differences in stem cytology;d8227 and four inbred lines with different backgrounds were used for genetic mating design to analyze dwarf culm.Inheritance mode of genes; construct a targeting population, perform high-throughput sequencing with BSA method, conduct preliminary mapping of dwarf genes, and perform allelic identification of dwarf materials with known positioning intervals to clarify target genes and known genes relationship. The results showed that the plant height of d8227 increased by 9.35%, the ear position increased by 31.50%, the leaves of d8227 decreased, and the length of stem nodes increased.dwarf-12 increased by 52.21%, 5.26%, 23.76%, 6.93%, 12.02%; using the paraffin section method, observe the characteristics of transverse and longitudinal cells on the upper, ear and lower ear of d8227 and dwarf-12 with a microscope, and d8227 was longitudinally sectioned.The cells were loosely arranged and the cells were obviously elongated;the dwarf-12 cells were arranged regularly and compactly.After measuring the cell area, ear and lower ear cell area of the d8227 were significantly increased than that of the dwarf-12, which was mainly caused by the elongation of the d8227 cells. Through genetic analysis, the dwarf stalk gene was a single recessive gene, and the gene was preliminarily located.The dwarf stalk gene were located at 190-215 Mb of chromosome 1.The dwarf maize that had been located in the interval was selected for allelic identification.The two-year planting results showed that d8227 and 123d, Na360 were not alleles, but may be alleles with 125d and 123d, which needed follow-up fine mapping and in-depth research. On the whole, d8227 is a medium dwarf material with excellent characters and has breeding potential, but further studies such as fine positioning are needed to judge its utilization value.

Tiller-related traits are important characteristics of wheat plant type,which determine plant structure and affect grain yield.In order to understand the inheritance and drought resistance of tiller-related traits in wheat under different water conditions,and to excavate the loci related to tiller-related traits,240 wheat varieties (lines) were selected as the subjects of this study,based on the phenotypic identification of tiller angle,effective tiller number and yield per unit area under normal irrigation (NI) and drought stress (DS) conditions,and the comprehensive evaluation of drought resistance,combined with 90K gene chip,genome-wide association study (GWAS) was to identify genetic loci for tiller-related traits and to screen for superior germplasm.The tiller angle,effective tiller number and yield per unit area showed significant difference,and the coefficient of variation ranged from 0.07 to 0.33.According to D-value,the drought resistance of Zhongyou 206 was the best.A total of 54 stable genetic loci significantly associated with tiller angle and other traits were detected, distributed on all chromosomes except 3D, 4D and 5D. Three identical stable loci were commonly detected under both treatments, located on chromosomes 2B, 4B, and 6B. Additionally, four pleiotropic loci were commonly detected in different traits, located on chromosomes 2B, 2D, and 5B.At the same time,the haplotype analysis of Ra_c491_902 (R²=5.45%—17.91%),which was significantly correlated with tiller angle on chromosome 2B,showed that there were three haplotypes:TA-Hap1,TA-Hap2 and TA-Hap3,the haplotypes (lines) containing TA-Hap1 were mainly derived from Huanghuai winter wheat regain.Five candidate genes related to tiller angle were screened by screening the stable genetic loci detected under different treatments.Gene annotation of the genes selected on Ra_c491_902 showed that the genes encoding cytochrome P450 family protein can be used as important genes such as regulating tillering angle,plant drought resistance and defense,to explore the association between genes and phenotype,and lay the foundation for the genetic improvement of tiller-related traits in wheat.

To reveal the mechanism of drought resistance of different resistant rice during germination period,Rice drought-sensitive materials(Calrose,Jingning 10,Shanxing 86)and drought resistance materials(Farry,Songjing 3,Ningjing 36)were studied on the effects of simulated drought stress(15% PEG-6000)on the growth index,physiological indexes and corresponding gene expression of different rice seeds.The results showed that under normal conditions,there were no significant differences in the expression levels of growth indicators and stress-related genes between drought-sensitive and drought-resistant cultivars.However,changes in physiological indicators were shown that there were no significant differences in the activities of superoxide dismutase(SOD) and peroxidase(POD),the contents of soluble sugar(SS) and hydrogen peroxide(H₂O₂) among different genotypes.The contents of malondialdehyde(MDA) and superoxide anion($\mathrm{O}_{2}^{\bar{.}}$) in the drought-sensitive cultivar Shanxing 86 were significantly higher than those in other materials,and the contents of catalase(CAT),proline(Pro) and soluble protein(SP) of drought resistant Ningjing 36 were significantly higher than those of other materials as well.Under drought stress,the relative germination potential(RGP),relative bud length(RSL),germination drought resistance index(GDRI)and vitality index(VI)of germinating seeds increased by 0.03—0.07 percentage,0.32—0.39 percentage,0.12—0.18 percentage and 92.41%—108.39%,respectively;MDA and reactive oxygen species($\mathrm{O}_{2}^{\bar{.}}$,H₂O₂) contents in germinating seeds of drought-resistant cultivars decreased by 2.54%—61.64%,19.60%—46.30% and 35.61%—62.02% respectively compared with drought-sensitive cultivars.The contents of osmotic regulating substances(Pro,SS,SP) increased by 5.93%—18.29%,1.08%—7.97% and 3.47%—6.03% respectively.The activities of antioxidant enzymes(SOD,POD, CAT) were increased by 17.29%—33.12%,15.24%—76.06% and 14.68%—18.61% respectively.The relative expression levels of OsP5CS,antioxidant enzyme synthesis genes (OsALM1, OsPOX1, OsCATC) were up-regulated by 2.66%—182.31% and 57.14%—513.27%,0.38%—109.06% and 63.39%—184.25% respectively.Comprehensive analysis showed that drought stress inhibited the germination of rice seeds and affected the physiological characteristics of seeds and the expression of corresponding genes during germination.Under drought stress,vigor index(VI),peroxidase(POD)and peroxidase synthesis gene(OsPOX1)are the key indicators affecting rice seed germination,whether it is drought-resistant or drought-sensitive materials.In addition to the above indicators,soluble protein(SP),proline synthesis gene(OsP5CS)and catalase gene(OsCATC)are other key indicators affecting drought-resistant materials.Relative shoot length(RSL),hydrogen peroxide(H₂O₂)and superoxide dismutase gene(OsALM1)are other key indicators affecting drought-sensitive materials.

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

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

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

NRL(NPH3/RPT2-Like)is a type of light-responsive protein unique to plants and plays a vital role in the phototropic signal pathway. To reveal the NRL gene maize genome's characteristics and expression,we analyzed them using bioinformatics methods combined with qRT-PCR technology. The property,structure,evolution of their encoded proteins,and growth period tissue expression and stress expression were analyzed. 31 ZmNRL genes identified were located in nine maize chromosomes,encoding protein amino acids 464-749 aa,which predicted to have chloroplast,nuclear and cytoplasmic locations. According to protein conservation,ZmNRL family was divided into four categories. Their gene structure also presented certain conservation,the most contained four exons. Analysis of the cis-elements of gene promoters revealed a large number of abscisic acids,jasmonic acid,light response,and anti-oxidation elements,among which G-box and Sp1 were two types of light-related elements. The expression of ZmNRL family genes in tissues during the growth period showed a temporal and spatial specificity,and the majority expression level was not high. Only ZmNRL2,ZmNRL4,ZmNRL24,and ZmNRL29 highly expressed. Furthermore,the characteristic modules were produced based on the data of the tissue co-expression genes. And the GO enrichment analysis of a particular leaf growth module containing six ZmNRL genes,mainly associated with the plastid organization biological processes and rRNA binding molecular functions. The expression of ZmNRL5,ZmNRL7,ZmNRL12,and ZmNRL19 genes were analyzed by qRT-PCR under salt,drought,high temperature,and Rhizoctonia solani inoculation treatments. The results showed that ZmNRL12 was significantly up-regulated in maize seedlings treated with high temperature,while ZmNRL5,ZmNRL7 and ZmNRL19 genes were down-regulated in drought,salt and pathogen treatments. In summary,31 ZmNRL genes were identified in the maize genome. They not only had apparent specific tissue expression but also participated in biotic and abiotic stress responses.

Leaf color mutant is one of the ideal materials to explore the development mechanism of chloroplast and the utilization of C4 light energy in foxtail millet. To study the molecular mechanism of leaf color mutant in foxtail millet,we screened and identified a stripe leaf mutant wsl2 from the EMS mutagenesis library of the main variety Changnong 35. The mutant was studied through phenotype identification,genetic background detection and genetic analysis,location of the mutant gene rapidly and precisely with MutMap method and developing co-isolated molecular markers according to the mutation site. The results showed that wsl2 showed stripe leaf phenotype at seedling stage,but returned to normal leaf phenotype from jointing stage. wsl2 had the same genetic background as the wild type by detecting the genetic background. And wsl2 was controlled by a recessive nuclear gene with genetic analysis. The association interval contained 9 non-synonymous mutant genes by MutMap method. Seita.9G561800 encoded a PsbP-like protein associated with chloroplast,containing 6 exons and 5 introns. A G/T base mutation occured at 77 bp in the first exon,resulting in an arginine(R)becoming leucine(L). The mutation site of candidate gene in wsl2 mutant was further verified by the dCAPS marker MRI498-1(Cac8 Ⅰ)and the co-isolation marker MRI501-3. This study identified a new stripe leaf mutant wsl2, which laid a theoretical foundation for further study on the mechanism of photosynthesis reaction of PsbP gene,enriched the resources of leaf color mutant,and verified the effectiveness of MutMap method in cloning mutant gene in foxtail millet.

To provide a solid foundation for studying the biological function of WRKY family members in kenaf in response to salt stress,all members of WRKY family were identified and their expression patterns were analyzed.Physical and chemical properties,phylogeny and conserved functional domains of WRKY gene family members were analyzed by bioinformatics method.The expression characteristics of WRKY gene family members under salt stress were analyzed by RT-PCR.The results showed that a total of 33 WRKY family members were identified,which were unevenly distributed on 12 chromosomes.There were certain differences in the physical and chemical properties of each member,such as amino acid number,molecular weight and theoretical isoelectric point.The conserved sequence WRKYGQK of each member did not change.Phylogenetic tree analysis showed that 33 WRKY family members were divided into 3 groups,GroupⅠ,GroupⅡ,Group Ⅲ,of which Group Ⅲ contained 5 subgroups.The real-time fluorescence quantitative RT-PCR results showed that there were 26 WRKY family members induced by salt stress,of which 23 had positive regulation and 3 had negative regulation.A total of 33 WRKY family members of kenaf were identified,of which 26 WRKY family members were involved in the salt stress response of kenaf.

WRKY is a unique class of transcription factors in plants,which plays an important role in plant abiotic stress response,seed dormancy and germination,growth and development,etc.In order to reveal the function and underlying molecular mechanism of GmWRKY44 gene in soybean WRKY transcription factor family,bioinformatics analysis and biology function verification of soybean Williams 82 GmWRKY44 were performed.GmWRKY44 gene was 1 077 bp in length and encoded 358 amino acids;the results of structural prediction and evolutionary analysis showed that,the secondary structure was composed of 23.46% α-helix,4.75% β-fold,58.94% irregular coil and 12.85% extended chain,and the tertiary structure was unified with the secondary structure;It contained a conserved WRKY domain, the zinc finger structure was of the C₂H₂ type, and it belonged to the WRKY IIc subfamily; GmWRKY44 is a homologous gene of Arabidopsis thaliana AtWRKY71 with a similarity of 35.56%, and the two genes had similar gene structures. RT-qPCR analysis showed that GmWRKY44 responded to salt stress and its expression level first decreased and then increased.Under salt stress,the germination rate and root length of wild-type(Col-0)and GmWRKY44 overexpressing Arabidopsis lines were inhibited to a certain extent,but GmWRKY44 overexpressing lines were significantly better than Col-0.In addition,under salt stress,the growth inhibition of GmWRKY44 overexpressing lines was lower than that of Col-0.Physiological index analysis revealed that under salt stress,the overexpression lines of GmWRKY44 exhibited significantly higher activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT)than Col-0,while the content of malondialdehyde(MDA)was significantly lower than Col-0.These data indicated that overexpression of GmWRKY44 could improve salt tolerance in transgenic Arabidopsis.

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 explore new approaches for high-oil peanut breeding and establish a new method for directly developing high-oil peanut germplasm,this study employed in vitro mutagenesis breeding technology to create new high-oil peanut germplasm.Jihua 9 embryo leaflet was used as mutagenic test materials,Jihua 9 and Jihua 54 were used as control test materials,and bleomycin was used as mutagenic agent.The ovules were sterilized and placed in gradient mutagenesis medium and screened for semi-lethal concentrations of bleomycin.After somatic embryos germinated into seedlings,sterile peanut seedlings were used as rootstocks,and transplanted to the field.Bioinformatics analysis of two known regulated peanut fat synthesis genes WRI1 and experimental feasibility validation by the correlation of WRI1 gene expression in grain and crude fat content of mutagenic plants were conducted.The results were best when the bleomycin was 3 mg/L.The crude fat content of IM13-3 was higher than that of Jihua 9(CK₁,test variety control)and Jihua 54(CK₂,high oil variety control).Two WRI1 genes,WRI1X2 and WRI1X1,encoding 366 and 357 amino acids,respectively,were both unstable hydrophilic proteins. WRI1 gene expression and crude fat content were significantly positively associated in grain.Bleomycin was first used as a peanut vitro mutagenesis agent,and IM13-3 was obtained with a crude fat content of 56.64%.It further proves the authenticity of Jihua 9 high oil mutant and the feasibility of peanut in vitro mutagenesis method. The gene expression level of the high-oil mutant WRI1 was determined and was significantly different from the control varieties. Demonstrate the feasibility of breeding methods for in vitro mutagenesis of peanut.

Serine hydroxymethyltransferase(SHMT),as an important enzyme involved in basic metabolism,plays an important role in plant cell metabolism,photorespiration and defense activities.To understand the bioinformatics function of the SHMT gene family in watermelon,explore its gene expression characteristics under abiotic stress,and provide a basis for the functional development of watermelon SHMT and the breeding of watermelon stress-resistance genes.Bioinformatics methods were used to identify SHMT family,and RT-qPCR was used to analyze the expression patterns of ClSHMTs in different tissues and abiotic stresses.The results showed that 8 ClSHMTs gene family members were identified in the whole genome of watermelon,which were unevenly distributed on 6 chromosomes and named ClSHMT1—ClSHMT8 in turn.There were some differences in the physical and chemical properties of each gene family member,such as the number of amino acids,molecular weight,isoelectric point.The protein contained 471—585 amino acids,with molecular weight of 51.87—65.00 ku and isoelectric point of 6.57—8.52,all of which were hydrophilic proteins.The subcellular localization prediction was mainly distributed on mitochondria.Gene structure and protein conserved motifs analysis showed that the ClSHMTs structure consisted of 4—15 exons and 3—14 introns,and all ClSHMTs contained conserved SHMT domains.Furtherly,phylogenetic analysis with 6 species such as cucumber and wheat showed that 50 SHMTs were divided into 3 sub-families,Group Ⅰ—Ⅲ.Promoter of ClSHMTs contained cis-acting elements related to light response,plant hormone response and stress response.The expression pattern analysis showed that 6 ClSHMTs were expressed in different tissues of watermelon,and the expression levels of ClSHMT1,ClSHMT4,ClSHMT5,ClSHMT8 in leaves were significantly higher than those in other tissues.Under low temperature,drought and salt stress,the expression abundance of ClSHMTs varies,but the expression was mainly up-regulated.In conclusion,this study systematically analyzed the SHMT gene family in watermelon,and will provide a reference for the further study of the biological functions of ClSHMTs.

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.

Phytosulfokine receptor(PSKR)plays an important role in promoting plant cell proliferation and is involved in plant response to abiotic stresses.To explore the sequence characteristics and the function of wheat PSKR genes,the cDNA sequences of three homologous genes of TaPSKR1 were cloned from wheat variety Jinmai 47 by homologous cloning technology,named TaPSKR1-6A,TaPSKR1-6B and TaPSKR1-6D because of their locations on chromosome 6A,6B and 6D,respectively.The gene structure,protein physical and chemical properties,cis acting elements,functional domains and evolutionary relationships were analyzed by bioinformatics analysis.The expression patterns of TaPSKR1 genes in different tissues and under different stresses were detected by qRT-PCR.The results showed that TaPSKR1-6A,TaPSKR1-6B and TaPSKR1-6D all contained one exon.The open reading frame(ORF)of the three TaPSKR1 genes were 3 153,3 132,3 156 bp,respectively,which encoded 1 050,1 043 and 1 051 amino acid residues.Bioinformatics analysis showed that TaPSKR1 proteins were located on the cell membrane,containing signal peptide,transmembrane domains,eight LRRs type domains and intracellular kinase domain,which belonged to PSKR gene family.Phylogenetic analysis showed that TaPSKR1 proteins had closely relationship with its related species and rice,which were clustered into the same subgroup.The results of expression analysis showed that TaPSKR1 genes were expressed in roots,stems,leaves and seeds,and the expression levels in roots were the highest.Under drought and salt stress treatments,the expressions of three homologous copies of TaPSKR1 genes were sharply upregulated in leaves,suggesting that TaPSKR1 might play an important regulatory role in wheat defense to abiotic stresses.

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.

To study the resistance mechanism of TaHis gene to Fusarium head blight(FHB)in wheat,the full-length coding sequence of TaHis was cloned,and the bait vector pGBKT7-TaHis was constructed,which was then used as bait for screening a yeast two-hybrid library of wheat ear induced by FHB.After obtaining the interacting proteins,yeast two-hybridization and bimolecular fluorescence complementation were further used to verify the interaction between these proteins,and RT-qPCR was used to analyze the expression pattern of TaHis interacting protein induced by FHB in resistant and susceptible cultivars respectively.The results showed that the bait vector pGBKT7-TaHis was successfully constructed,and 18 yeast monoclones were obtained on the four deficient selection medium(SD/-Leu/-Trp/-His/-Ade)after yeast two-hybrid library screening.Blast analysis showed that a total of 5 proteins were obtained,and the coding sequence of serine/arginine-rich mRNA splicing factor SR45a-like(TaSR)was identified in 6 colonies.We cloned the full-length coding region of TaSR gene from Bainong 4299 and constructed pGADT7-TaSR vector.The experiment of yeast two-hybrid showed that the yeast cells co-transformed with pGADT7-TaSR and pGBKT7-TaHis grew well and appeared blue on SD/-Leu/-Trp/-His/-Ade/ X-α-Gal/AbA,indicating that TaSR and TaHis directly interacted in yeast cells.The vectors of YC-TaHis and YN-TaSR were constructed,and the bimolecular fluorescence complementary experiments were performed.The results showed that strong fluorescence signals were generated in tobacco cells co-transferred with YC-TaHis and YN-TaSR,which further verified the interaction between TaSR and TaHis.RT-qPCR analysis of TaSR gene expression showed that TaSR expression was up-regulated in resistant cultivar Bainong 4299,while down-regulated in susceptible cultivar Bainong 607 upon FHB infection,indicating a positive correlation between TaSR expression level and FHB resistance in wheat.To sum up,the interaction between wheat TaSR and TaHis was proved,and TaSR expression level was positively correlated with FHB resistance in wheat.

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

In order to study the function of GhAOP2-like in cotton,based on the previous proteomic data from the laboratory of Institute of Cotton, Hebei Academy of Agriculture and Forestry Sciences, this study obtained the gene sequence of GhAOP2-like using homologous cloning strategy.The physicochemical properties,structure and subcellular localization of GhAOP2-like protein were analyzed by bioinformatics method.The tissue-specific expression of GhAOP2-like and the change of expression level under drought,salt and hormone treatment were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The results showed that the CDS sequence length of GhAOP2-like was 972 bp and encoded 323 amino acids.GhAOP2-like was located on chromosome D13.GhAOP2-like protein did not contain signal peptides and transmembrane domains,and was located in the cytoplasm with the molecular formula of C₁₆₅₄H₂₅₂₅N₄₂₉O₄₇₈S₁₉ and the theoretical isoelectric point was 5.20.The clustering analysis results showed that the AOP sequences in cotton and other species were clearly divided into two groups,but had the closest relationship with the AOP sequences of Hibiscus syriacus.qRT-PCR results showed that GhAOP2-like was expressed in roots,stems,leaves,and developing seeds,but the expression level of GhAOP2-like was the highest in roots.GhAOP2-like was up-regulated under drought,salt,GA₃,MeJA and ABA treatments,but the expression level changed most in MeJA treatment,indicating that GhAOP2-like might improve the resistance of upland cotton by participating in JA signaling pathway.

Abstract: Tetraploid wheat is the ancestor specie of common wheat and an important food crop.Aiming to provide new resistance sources for wheat variety breeding,the resistance tetraploid wheat germplasm was explored and their resistance genes were identified.TDI-1 is a cultivated emmer wheat that has been immune to powdery mildew in the field for many years.To determine the resistance genes carried by TDI-1,and provide a theoretical basis for genetic improvement of wheat resistance,a durum wheat TDU-1 that was susceptible to powdery mildew was used to hybridize with TDI-1,and their F₁ plants,F₂ population,and F_2:3 lines were obtained.Genetic analysis of resistance was conducted on parents TDI-1,TDU-1,and their hybrid offspring that were inoculated with powdery mildew isolate E09.Then,bulked segregant analysis method combined with molecular markers was used to map the resistance gene.The results showed that TDI-1 was susceptible to E09 during the seedling stage but immune during the adult stage.F₁ plants derived from the cross of TDI-1 and TDU-1 were immune to E09 during the adult stage.The resistance of adult F₂ individuals was separated,and the ratio of resistant and susceptible plants was 3:1($χ_{3:1}^{2}$=0.11,P=0.74);the ratio of the number of homozygous resistant,separated resistant,and homozygous susceptible F_2:3 lines was 1:2:1($χ_{1:2:1}^{2}$=0.47,P=0.79),indicating that the resistance to powdery mildew in the adult stage of TDI-1 was controlled by one dominant gene,temporarily named PmTDI-1.Subsequently,a set of molecular markers was used to amplify the parents and their F₂ population,and then four markers on chromosome 2A,including Xwmc407,NRM-2AS29,NRM-2AS45 and NRM-2AS84, confirmed to be linked to PmTDI-1. PmTDI-1 was between the flanking markers NRM-2AS45 and NRM-2AS84,with genetics distances of 1.8 cM and 4.6 cM,respectively.Therefore,the adult stage powdery mildew resistance gene PmTDI-1 was preliminarily localized on chromosome 2A.This study identified a novel dominant adult-plant-resistance powdery mildew gene PmTDI-1 from tetraploid wheat TDI-1.