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.

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.

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.

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.

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.

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

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

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

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

Hydrangea macrophylla is an ornamental plant with high aluminum(Al) tolerance and strong Al accumulation capacity.Its flower color is easily influenced by soil pH and the Al³⁺ content in sepals,yet the mechanisms underlying its Al tolerance remain poorly understood.The MYB transcription factor family plays a crucial role in plant stress responses.To investigate the function of MYB transcription factors in Al tolerance in H.macrophylla,this study used H.macrophylla 'Endless Summer' as experimental material.Gene cloning,bioinformatics,co-expression network,expression pattern analysis and yeast functional validation of HmMYB73 were conducted.The results showed that HmMYB73 encoded 266 amino acids,contained a typical R2R3 domain,and belonged to the R2R3-MYB subfamily.Genes co-expressed with HmMYB73 were involved in biological functions such as catalytic activity,transporter activity,response to stimulus and detoxification.Under Al treatment,HmMYB73 was significantly upregulated in the roots,leaves,and sepals of H.macrophylla,with the highest expression level observed in the roots,which was 16 times that of the control.Furthermore,overexpression of HmMYB73 significantly enhanced yeast tolerance to aluminum stress.In summary,HmMYB73 may be involved in the biological regulation during aluminum stress in H.macrophylla.

During the reproductive phase of Uncaria rhynchophylla,stem hooks undergo conversion into peduncles,consequently diminishing stem hook yield.To identify key regulatory factors governing this organ homology shift between stem hooks and peduncles in U.rhynchophylla,tissues from stem hooks and flowers were utilized to isolate its floral meristem gene,UrAP1(APETALA1).Comprehensive bioinformatic characterization was subsequently performed to elucidate the physicochemical properties of its encoded product.Concurrently,qRT-PCR analysis was conducted to quantifiy the relative transcript abundance of UrAP1 across distinct tissues of vegetative and reproductive branches,aiming to clarify its functional role and regulatory network in floral organogenesis.Results demonstrated that the UrAP1 coding sequence (CDS) spanned 729 bp,encoding a 242 amino acid polypeptide.This sequence exhibited substantial homology(94% identity) with the AP1 gene from Cephalanthus occidentalis and harbored conserved MADS-box and K-box domains.Furthermore,the UrAP1 protein lacked transmembrane domains,with primary localization predicted within the nucleus.UrAP1 expression was detectable and relatively stable across leaves,stems,stem nodes,and stem hooks of vegetative shoots.In contrast,its transcript levels varied significantly in reproductive shoots,showing distinct abundances in leaves,stems,stem nodes,peduncles,and flower buds.Expression peaked in young buds during early flowering stages and subsequently declined as buds matured.Comparison of the expression profiling between the two growth phases suggests a crucial role of UrAP1 in regulating floral bud differentiation in U.rhynchophylla.

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.

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.

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

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

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

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.

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

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

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.

In order to explore the relationship between MrAGL8 gene and pod dehiscence traits in Medicago ruthenica. In this study,Medicago ruthenica was used as the plant material.The MrAGL8 gene was amplified by PCR,cloned,and sequenced.Additionally,bioinformatics analysis,subcellular localization,and expression analysis in different tissues and organs were performed for this gene.The results showed that the complete coding region of MrAGL8 cDNA with a length of 711 bp was obtained through cloning using PCR amplification technology.Bioinformatics analysis results showed that MrAGL8 encoded 236 amino acids,it had MADS-box and K-box protein conserve domains.Its molecular weight was 27.39 ku,the theoretical isoelectric point was 8.69,the total number of positively charged residues was 39,the total number of negatively charged residues was 36,and the instability coefficient was 48.5.The secondary structure of the protein contained α-helix and β-sheet.It was an unstable protein and belonged to a hydrophilic alkaline protein.Subcellular localization results showed that MrAGL8 protein was located in the nucleus.The expression analysis of different tissues and organs showed that the expression level of MrAGL8 in different tissues was stem>root>pod>leaf>flower,and the expression levels of roots and stems were significantly different from those of leaves,flowers and pods.The results showed that MrAGL8 gene was related to pod dehiscence.

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

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.

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

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.

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

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.

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

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

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

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

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

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

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

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

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

To understand the changes in endogenous hormones and related gene expression levels after konjac infection with southern blight disease and reveal the main hormone pathways involved in konjac's response to southern blight disease,the ultra performance liquid chromatography (UPLC) and tandem mass spectrometry (MS/MS) was employed to detect the changes in the contents of phytohormone (auxin,abscisic acid,trans-zeatin nucleoside,jasmonic acid,and salicylic acid) in the three-month-old Amorphophallus muelleri,which was infected with southern blight disease for 0,1,3 and 6 days.Moreover,the gene expression levels of abscisic acid,jasmonic acid,and salicylic acid pathways was analyzed by Real-time Quantitative PCR.The results showed that southern blight disease caused changes in the levels of various endogenous hormones in konjac.The content of indole-3-acetic acid tended to increased and then decreased with the southern blight disease infection,while the content of indole-3-butyric acid showed tended to decrease;the content of trans-zeatin nucleoside significant decreased with the infection of southern blight disease;the content of abscisic acid showed a significant increase followed by a decrease with the infection of southern blight disease;the content of jasmonic acid metabolites and salicylic acid metabolites were significantly increased than the control group with the infection of southern blight disease.The content of jasmonic acid of konjac infected with southern blight disease 1,3,and 6 d was 2.31,2.31,and 5.08 times that of the control group,and the content of salicylic acid was 5.53,4.60,and 7.38 times that of the control group,respectively.Eight genes related to abscisic acid,jasmonic acid,and salicylic acid pathways were activated and participated in the response process of konjac to southern blight disease.The above results indicated that the endogenous hormone homeostasis was disrupted after konjac infection with southern blight disease,activated the expression of plant hormone pathway related genes,and jasmonic acid and salicylic acid might play an important role in konjac's resistance to southern blight disease.

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

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

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

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

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

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

Low temperature is the main factor affecting the distribution of Camellia in Northern China.In order to further expand the distribution range of Camellia and enhance the diversity of garden plants in Northern China.The yeast library was constructed by Gateway recombination technology using the leaves of C.japonica(Naidong)Daochengchunzao as materials.The total number of clones in the primary library was 1.44×10⁷ cfu,the total number of clones in the secondary library was 1.12×10⁷ cfu,the positive rate of recombination was 100%,the titer of yeast library was 1.00×10⁸ cfu/mL,and the average insert fragment of yeast clone was more than 1 000 bp,which met the standard of library construction.The bait vector pGBKT7-CjCBF1 was constructed by double enzyme digestion and homologous recombination,which was non-toxicity and self-activation activity in yeast cells.The library screening was conducted by plasmid cotransfer method,and 46 candidate proteins interacting with CjCBF1 were obtained,whose functions involved plant growth and development,flowering and fruiting,and response to stress,etc.CjRAV1 was selected as a candidate protein.Primers were designed according to the transcriptome database of C.japonica(Naidong)and Camellia sinensis genome database.The CjRAV1 gene was cloned and the full length of the gene CDS was 1 023 bp.Bioinformatics results showed that the gene encoded 341 amino acids with a relative molecularity of 37.99 ku,a protein isoelectric point of 9.10,an instability index of 34.74 and a lipid index of 76.60.The amino acid sequence alignment and phylogenetic tree construction of the species with close homology to CjRAV1 were carried out.It was found that it was closely related to Camellia lanceoleosa,Diospyros lotus and Actinidia chinensis.In order to reduce the false positive probability of the library screening,the pGADT7-CjRAV1 vector was constructed and verified with pGBKT7-CjCBF1 by point-to-point.It was confirmed that there was an interaction between the two proteins,which laid a foundation for further study on the molecular mechanism of low temperature response of C.japonica(Naidong).

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

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

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

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

To explore the function of key genes in photosynthesis, a functional knockout mutant (zmC₄nadp-me) of ZmC₄NADP-ME, the gene encoding the rate-limiting enzyme of the dark reaction of photosynthesis in maize, was obtained. Evolutionary tree analysis showed that ZmC₄NADP-ME and its homologous genes exist in multiple copies in most plants, with diverse expression patterns. Phenotypic analysis revealed that the entire zmC₄nadp-me plant was yellow-green, and its seedling-stage leaves dried up and died rapidly under light. Chlorophyll fluorescence analysis indicated that Y(Ⅱ) and electron transport rate ETR(Ⅱ) of photosystem Ⅱ (PSⅡ) in zmC₄nadp-me decreased significantly, with little change in Y(NPQ), while the Y(NO) increased notably. Measurement of the absorption capacity (P700) of photosystem Ⅰ (PSⅠ) found that both the electron transport rate (ETR(Ⅰ)) and the actual photoelectron efficiency (Y(Ⅰ)) of zmC₄nadp-me dropped substantially, and the gap widened with increasing light intensity. Under specific light intensities, Y(ND) and Y(NA) of zmC₄nadp-me were greater than those of the wild type (WT). In conclusion, ZmC₄NADP-ME is essential for plant growth and development. Disruption of this gene severely stresses PSⅡ, and the plant can't alleviate this stress by increasing Y(NPQ). Meanwhile, at low light intensities, the inhibition of PSⅠ may originate from the electron donor side of PSⅠ, and as the light intensity increases, the inhibition from the electron acceptor side of PSⅠ becomes a key factor.