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.

Cabbage fusarium wilt is a serious soil-borne fungal disease caused by Fusarium oxysporum f.sp.conglutinans (FOC),which affects the yield and quality of cabbage.In order to clarify the biological function of the transcription factor SNT2 in this pathogen,the SNT2 gene knockout mutant ΔSNT2 in F.oxysporum was successfully obtained by homologous recombination and protoplast transformation,and its phenotype and pathogenicity were analyzed.The results showed that SNT2 in F.oxysporum encoded 1 529 amino acids,had a SANT domain that binds to DNA,and the protein belonged to a hydrophilic protein.Compared with the wild type strain,the mycelial growth rate of the ΔSNT2 mutant decreased and the septation increased significantly,and the conidial production decreased significantly.Based on the results of exogenous stress,ΔSNT2 was insensitive to osmotic stress of 1 mol/L sorbitol,but its tolerance to oxygen stress,salt stress and cell wall stress of 0.1% H₂O₂,2 mol/L NaCl and 0.05% Congo red was reduced.At the same time,the pathogenicity test showed that the disease index of ΔSNT2 mutant was significantly lower than that of wild type,and the deletion of SNT2 resulted in a significant decrease in the pathogenicity of F.oxysporum.In conclusion,the transcription factor SNT2 plays an important role in maintaining the integrity of the cell wall during the interaction between the pathogen and the host,and participates in the regulation of the growth and development of F.oxysporum and the expression of pathogenicity.

In order to accelerate the breeding of pink-tomato varieties,a rapid breeding strategy was used to generate pink-fruited tomato material by CRISPR/Cas9-mediated gene editing of SlMYB12.Two adjacent target sites within the first exon of SlMYB12 were selected to construct the CRISPR/Cas9 binary vector, and then the vector was introduced into red-fruited inbred line R18-10C through Agrobacterium-mediated transformation. The homozygous mutants without exogenous Cas9 were screened using specific primers and their agronomic traits and fruit nutritional quality were analyzed. Sequencing results showed that three homozygous mutants of different mutation types were obtained and all of which were frame shift mutations caused by base deletion. Compared with wild-type red fruit tomatoes, SlMYB12-edited plants grew and developed normally,and there were no significant difference in plant height,single-fruit weight,total yield per plant,fruit total soluble solid content and lycopene level,but the mature fruit showed pink and the flavonoid naringenin chalcone(NarCh)content of tomato peels was significantly reduced in MYB12-edited plants.In summary,the procedure for the generation of pink-fruited tomato plants through CRISPR/Cas9-mediated targeted mutagenesis of SlMYB12 was set up and new pink-fruited tomato germplasms with stable inheritance were obtained.

PHR1 is a crucial factor in balancing plant disease resistance and low phosphorus stress resistance.To investigate the nature and function of the StPHR1 gene in potato and to explore the role of StPHR1 in the process of potato resistance to Alternaria solani infection,the CDS sequence of the StPHR1 gene was cloned by PCR technology using potatoes as the research material,and the structural,physicochemical properties,and phylogenetic relationships of StPHR1 were analyzed and predicted using bioinformatics software,then,the expression level of StPHR1 during the infection of potatoes by A.solani and under different hormone treatments was analyzed using qRT-PCR technology,and subcellular localization analysis of the protein was conducted using laser confocal microscopy technology.The results showed that the CDS of the StPHR1 gene was 1 353 bp,encoding 450 amino acids.The protein had a molecular formula of C₂₁₄₇H₃₃₉₉N₅₉₅O₇₁₁S₁₈,a molecular weight of 49.51 ku,and a theoretical isoelectric point of 5.07,encoding a hydrophilic,unstable protein with no signal peptide and no transmembrane structure.Its secondary structure consisted mainly of random coil and α-helix.Phylogenetic tree analysis revealed that the StPHR1 protein was most closely related to Arabidopsis thaliana; conservative domain analysis revealed that the StPHR1 protein,like other PHR1s,possesses both MYB-CC and MYB conserved structural domains at its C-terminus.Relative expression analysis found that StPHR1 was significantly induced by A.solani and salicylic acid,and it was hypothesized that StPHR1 played an important role in A.solani infection of potato and in the response to salicylic acid; and the subcellular localization showed that the StPHR1 protein was localized in the nucleus.It is hypothesized that StPHR1 may regulate potato resistance to A.solani through its MYB transcription factor activity and response to salicylic acid.

The naked seeds of zucchinii possess significant natural advantages in processing.To investigate the genetic mechanisms underlying the hull-less trait in zucchini,we utilized the hulled 17pu10 (P₁) and the hull-less 17pu08 (P₂) as parental strains,and constructed populations of F₁ (P₁×P₂),F₂,and BC₁.The phenotypes of the zucchini seeds in the progeny population were assessed.The results showed that the number ratio of hulled seeds and hull-less seeds in the progeny population was in line with 3∶1 separation ratio.This indicated that the hull-less trait was regulated by a single gene and that the hull-less allele was recessive.Genetic mapping within this interval revealed that the gene for hull-less in zucchini was located between the markers InDel3157329 and InDel3724121,with genetic distances of 1.4,2.6 cM,respectively,while the physical distance was 0.6 Mb.The annotation and function analysis of 24 genes in the interval showed that 4 genes were directly or indirectly involved in the biosynthesis of cell wall,cellulose and lignin.Further analysis of the expression differences of 4 genes showed that only Cp4.1LG12g04350 and Cp4.1LG12g04370 had significant differences in expression levels during seed development.It was inferred that Cp4.1LG12g04350 or Cp4.1LG12g04370 was the candidate gene controlling the hull-less trait.In addition,InDel markers linked to hull-less genes were developed,which could be used as markers to identify zucchini hull-less traits,so as to accelerate the breeding of high-quality zucchini seed hull-less varieties.

Cucumber powdery mildew is one of the main diseases that adversely impacts cucumber production,posing a significant challenge to its sustainable cultivation.Identifying genes related to cucumber resistance to powdery mildew can help understand the genetic principles and molecular mechanisms of cucumber resistance to powdery mildew,and provide diverse gene resources for disease resistant breeding.This study constructed F₁ and F₂ populations of QK×QG using cucumber resistant inbred line QK and susceptible inbred line QG as parents.Using the extreme trait mixed pool resequencing (BSA-seq) method,the genomic regions harboring the resistance genes of cucumber powdery mildew were preliminarily located.By integrating transcriptome data with gene annotation information,the association interval of the disease phenotype was narrowed,sequence variations were identified,and key genes were screened.The results showed that the resistance of powdery mildew may be controlled by recessive genes,and the population of F₂ showed a continuous normal distribution from resistant to susceptible.The BSA-seq analysis,combining the SNP-Index method and QTG(quantitative trait genomics)-seq method analysis highlighted the 19—21 Mb region of chromosome 5,where there were 77 annotated genes with SNP differences between samples,including 33 non-synonymous mutations.The transcriptome sequencing (RNA-seq) results showed that there were 309 upregulated genes and 697 downregulated genes in the susceptible material.The expression levels of 13 genes within the candidate segment of chromosome 5 showed significant differences after infection.Through a comprehensive analysis of differentially expressed genes and BSA,the candidate genes in this segment were narrowed to 3,and only SNP mutations were detected in the LOC101207011 gene.The candidate gene LOC101207011 was characterized by a mutation resulting in an amino acid change from Valine at position 656 to Leucine.This gene emerges as the primary focus of our investigation due to its potential role in conferring resistance to powdery mildew.

In order to reveal the expression patterns of PsbS protein in Nicotiana tabacum,the full-length NtPsbS gene sequence was cloned from cDNA of the tobacco cultivar K326,and the multiple protein sequences of NtPsbS gene and PsbS genes of seven crops including rice,tomato and soybean were compared by DNAMAN software.The phylogenetic analysis was carried out by MEGA 11 software,and the phylogenetic tree was established by neighbor-joining method.Tissue expression of NtPsbS gene in tobacco at different growth stages was detected by qRT-PCR.The plant expression vector pS1300-PsbS-GFP was constructed to find subcellular localization of the NtPsbS mature protein.Finally,the mRNA levels of NtPsbS under abiotic stresses were analyzed in K326.The results showed that the total length of tobacco NtPsbS gene was 825 bp,encoding 274 amino acids.The homology of tobacco NtPsbS protein and tomato SlPsbS protein was the highest up to 91%.The NtPsbS gene exhibited the highest mRNA levels in the leaves among the tissues,including leaves,roots,stems,seeds and other parts of K326.The mature NtPsbS protein was located in chloroplasts.For the abiotic stress,it was found that the expression level of NtPsbS was significantly increased under salt stress,cold stress and Abscisic acid(ABA)treatments.In summary,the expression level of NtPsbS was the highest in tobacco leaves at different growth stages,and it was higher when under the treatments of salt stress,cold stress and ABA,indicating that this gene may be involved in the salt and cold stress resistance and ABA metabolic pathway in tobacco,which could provide basis and reference for the functional analysis of the NtPsbS gene in future.

To reveal the sequence characteristics,subcellular localization,and expression pattern of PatASIL2,belonging to the Trihelix transcription factor family,the PatASIL2 gene was first cloned using the cDNA of Pogostemon cablin (patchouli)as a template,and the bioinformatics analysis was further performed.The PatASIL2-EGFP expressed vector was constructed and transformed into Arabidopsis protoplasm to investigate the subcellular localization of PatASIL2.The real-time quantitative reverse transcription PCR(qRT-PCR)was also performed to detect the expression profiles of PatASIL2 gene in different tissues of patchouli and under treatments of methyl jasmonate(MeJA),salt stress,drought stress,and cold stress.The results indicated that PatASIL2 gene contained an open reading frame of 1 035 bp and encoded 344 amino acids.The PatASIL2 protein was an unstable hydrophilic protein without transmembrane domains and signal peptides and had 41 serine phosphorylation sites and one Myb_DNA-bind_4 conserved domain.The phylogenetic analysis showed that PatASIL2 was classified into the SIP1 subfamily of Trihelix transcription factor family,and clustered closely with Sesamum indicum SiASIL2.The subcellular localization result indicated that PatASIL2 was a nuclear-localized protein.The qRT-PCR results indicated that PatASIL2 expressed in the young leaf,mature leaf,old leaf,stem,and root in patchouli,particularly with the highest expression in the old leaf.The PatASIL2 expression was significantly upregulated by MeJA at 12—24 h and salt stress at 3—24 h.The expression of PatASIL2 was significantly upregulated at 24 h after drought treatment,and at 12 h after cold treatment.

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

The aim was to study the biological function of FolSid1 gene in Fusarium oxysporum f.sp.lini and its protein localization in Fusarium by cloning the gene.The gene sequence of FolSid1 was cloned by homologous comparison with F.oxysporum, and based on the principle of homologous recombination, a gene deletion box containing hydromycin resistance gene(hph)was constructed by Split Marker strategy,and the gene deletion mutant(ΔFolSid1)was obtained by PEG-mediated transfer into protoplasts of the wild type.pZESH1,a green fluorescent expression vector containing FolSid1 gene was constructed,and the subcellular localization of FolSid1-EGFP fusion protein was performed.The results showed that the sequence of FolSid1 gene consisted of 5 392 bp,which contained 3 introns.Compared with the wild type and the external insertion mutant, the knockout mutant ΔFolSid1 conidia showed a significant decrease in yield, although they did not differ in morphology and size; morphological observations revealed that the growth rate of colonies from the knockout mutant was significantly slower. The experiments of subcellular localization showed that FolSid1 protein was located in the cell membrane of mycelia cell.FolSid1 gene regulated the vegetative growth of mycelium,conidiogenesis and pathogenicity of Fusarium oxysporum f.sp.lini.

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

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

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

To investigate the function of lysophosphatidicacid acyltransferase 2(LPAT2)in Brassica napus,from which one copy(A07)of BnaLPAT2 was cloned by PCR.we constructed the overexpression vectors p35S∷BnaLPAT2-A07 and the seed-specific expression vector pNapin∷BnaLPAT2-A07,and by utilizing Agrobacterium-mediated genetic transformation method,obtained a total of 15 and 11 transgenic Brassica napus cv.Zhongshuang 6 respectively by PCR positive detection.Real-time Quantitative PCR(qRT-PCR)showed that the transcript levels of BnaLPAT2-A07 in most tissues of T₃ overexpressed rapeseed were higher than that of CK.However,in the seed-specific expression tissues of T₃ transgenic rapeseed,the BnaLPAT2-A07 genes were strongly expressed in the development and maturation stages of silique.Soxhlet extraction results showed that the oil content in the transgenic seeds driven by the 35S or Napin promoter accumulated 1.39 and 2.36 percentage point more oil than control seeds,respectively.The fatty acid components of transgenic rape were detected by gas chromatography.Compared with CK,the content of linolenic acid was increased by 3.13 and 1.47 percentage point,respectively.Taken together,the BnaLPAT2-A07 could promote seed oil synthesis,however,the specific selection function of BnaLPAT2-A07 for linolenic acid needs to be further verified.

The identification and expression pattern analysis of CmPIPs gene family in melon could provide theoretical basis and support for further exploring the function of CmPIPs gene family and the genetic improvement of melon.TBtools,MEME,MEGA X and Plant-CARE tools were used to analyze the bioinformatics of CmPIPs,and the expression level of CmPIP2;7 in the pericarp of melon at different stages after pollination,and the expression level of each member of CmPIPs in different tissues and different concentrations of plant hormone treated young leaves were visualized in the software GraphPad Prism 10.The results showed that CmPIP2;7 and CsPIP2;8 had nearest kinship;the 12 members of the CmPIPs family were mainly distributed on chromosomes 1,3,4,5,9,10 and 11;except that CmPIP2;8 had 3 CDS regions,the other members had 4 CDS regions.The promoter regions of each member of CmPIPs had multiple cis-acting elements,hormone responsive elements,such as auxin,gibberellin,and abscisic acid.The expression level of CmPIP2;7 was significantly up-regulated during rapid development and maturity of melon fruit.Members of various families of CmPIPs were expressed in different tissues of melon.After treatment with 40.0 μmol/L auxin,the expression level of CmPIP2;4 was significantly up-regulated,while the expression level of CmPIP1;1,CmPIP 2;1,CmPIP2;2 and CmPIP2;3 were extremely significantly down-regulated,and when the concentrations of abscisic acid were 0.4,4.0 and 40.0 μmol/L,the expression level of CmPIP1;1,CmPIP2;1,CmPIP2;3,CmPIP2;9 were significantly down-regulated.After treatment with 44.640 μmol/L methyl jasmonate,the expression level of CmPIP2;1 and CmPIP2;5 were significantly down-regulated,while the expression level of CmPIP2;2,CmPIP2;3,CmPIP2;7 and CmPIP2;9 were significantly up-regulated.When the concentration of ethylene glycol was 4.0 mmol/L,the expression level of each member of CmPIPs was significantly up-regulated.The gene structure,sequence characteristics,evolutionary relationship and collinearity of CmPIPs gene family members were clarified,and their expression pattern were analyzed.

As one of the subfamily members of Ca²⁺-ATPase,ACA exerts a vital role in sustaining the intracellular Ca²⁺ concentration equilibrium and modulating plant growth and development in response to abiotic stress.To acquire an in-depth comprehension of the function of the lettuce ACA gene family,bioinformatics methodologies were utilized to identify and dissect the members of the lettuce ACA gene family.The outcomes manifested that 17 ACA genes were identified in lettuce,denominated as LsACA1 to LsACA17;the LsACA genes were unevenly distributed across 8 chromosomes;the subcellular localization prediction results disclosed that all LsACA proteins were localized in the plasma membrane;the number of introns exhibited significant variation(0—32)among the members of the LsACA gene family,a total of 15 conserved domains of LsACA proteins were identified,with amino acids ranging from 21 to 50;the proportion of the secondary structure was alpha helix>random coil>extended strand>beta turn;based on phylogenetic analysis,LsACA proteins were categorized into 5 subfamilies,namely Group Ⅰ to Group Ⅴ;according to collinearity analysis,it was found that there were fragment duplications in 6 pairs of genes,the Ka/Ks of their collinear gene pairs were all less than 1,signifying that purifying selection was the preponderant force in evolution.The expression patterns of LsACA gene family members under different calcium ion concentrations were analyzed by qRT-PCR.The results showed that:compared with the control,the expression levels of 12 LsACA genes in the calcium-sensitive variety Baoshilü under low calcium treatment were extremely significantly downregulated,while the expression levels of 9 LsACA genes in the calcium-insensitive variety Yeluo were extremely significantly upregulated and the expression level of 1 LsACA gene was significantly upregulated.The ACA gene family members of lettuce were identified and analyzed,and the characteristics of the LsACA gene family members were revealed.

Chalcone synthase(CHS)is an important structural gene that regulates the early biosynthesis of flavonoid pathways in plants,and plays a role in plant growth and development and stress response.Previously,the key genes StCHS4 and StCHS5 for anthocyanin biosynthesis were identified in the potato CHS family by expression analysis.To further explore the function of potato StCHS4 and StCHS5 in the biosynthesis of flavonoids and anthocyanins,the characterization of StCHS4 and StCHS5 proteins was analyzed by online website.The 35S∷StCHS4-GFP and 35S∷StCHS5-GFP recombinant vectors were constructed by homologous recombination method based on the pRI101 binary vector,and then were transformed into Agrobacterium GV3101 strain.The subcellular localization of StCHS4 and StCHS5 proteins was determined by transient transformation of Nicotiana benthamiana.N.tabacum was used as the experimental material for transient overexpression and stable genetic transformation to analyze the content of total flavonoids and anthocyanins after overexpression of StCHS4 and StCHS5 genes.The results showed that the secondary structures of StCHS4 and StCHS5 proteins were mainly α-helix and random coil.StCHS4 was an unstable hydrophilic protein,and StCHS5 was a stable hydrophilic protein.The sequence alignment revealed that StCHS4 and StCHS5 had a close relationship with the CHSs of Capsicum annuum and Solanum lycopersicum,respectively.The results of subcellular localization revealed that StCHS4 and StCHS5 proteins were localized in the cytoplasm and cell membrane.In transient overexpression of tobacco,StCHS4 and StCHS5 genes significantly enhanced anthocyanin accumulation at 3—5 days after injection.Three transgenic N.tabacum lines of StCHS4 and StCHS5 gene were obtained,respectively.Compared with the wild type,the expression of StCHS4 and StCHS5 in transgenic plants was significantly higher,and the contents of total flavonoids and total anthocyanins were higher than those in the wild type.The total flavonoid content in StCHS4-OE3 and StCHS5-OE1 transgenic plants was significantly increased.The anthocyanin content in StCHS5-OE1 and StCHS5-OE2 plants increased by 89%,131%,respectively.The above results demonstrated that StCHS4 and StCHS5 were the key CHS genes in the flavonoid pathway of Solanum tuberosum,and the overexpression of StCHS4 and StCHS5 contributed to the biosynthesis of anthocyanins and flavonoids.

DNA damage significantly hinders impact plant growth and development.NBS1 plays an important role in the cellular response to DNA damage,to analyze the functional difference between NBS1 and its alternative splicing isoform,NBS1-3.Specific primers were designed to clone NBS1 and NBS1-3 from the complementary DNA of Arabidopsis thaliana,and the sequences of the NBS1,NBS1-3 and the three-dimensional structure of the proteins were analyzed.Transgenic Arabidopsis thaliana lines overexpressing of NBS1 and NBS1-3 were generated respectively,and nbs1 homozygous mutant plants were identified,detecting the expression levels of NBS1 in overexpressed lines NBS1 and NBS1-3.To further elucidate the functional distinction between NBS1 and NBS1-3,wild type,mutant and overexpressed plants were treated with 0.6 mmol/L Methyl Methanesulfonate(MMS)and observed damage area.The results of quantitative showed that the expression levels of NBS1 in overexpressed lines NBS1 and NBS1-3 were higher than wild type.PI staining of root tips showed that after 0.6 mmol/L MMS treatment,the mutant showed the highest relative damage area,while the NBS1-3 overexpressed lines exhibited the least damage,followed by NBS1 overexpressed lines and wild type.These results suggest a potentially more important role of NBS1-3 in DNA damage repair compared to NBS1.

To understand the member characteristics of the WRKY transcription factor family in hydrangea and their roles in response to leaf spot disease,this study utilized the bioinformatics method to characterize WRKY family members in Hydrangea macrophylla Endless Summer,and systematically analyzed the protein physicochemical features,gene structure,systematic evolution,collinearity and expression patterns under the infection with Corynespora cassiicola of WRKY family members.The results showed that there were 84 non-redundant HmWRKY members in H.macrophylla genome.All of the HmWRKYs were hydrophilic proteins and unevenly distributed on 18 chromosomes of H.macrophylla,encoding 112—1 046 amino acids.In addition,HmWRKY members were divided into 3 subgroups(Group Ⅰ—Group Ⅲ),which contained a conserved DNA binding domain composed of WRKYGQK and C2H2.The sequence length of HmWRKY varies greatly from 512 bp to 40 338 bp,and 8 collinear gene pairs with Ka/Ks ratios less than 1 were detected,indicating that the HmWRKY family experienced purification selection in evolution.18 HmWRKY members showed significantly differential expression after infection with C.cassiicola,of which 9 up-regulated and 9 down-regulated.The results indicated that these HmWRKY genes might play important roles in response to leaf spot disease of hydrangea.

4-coumaric acid:coenzyme A ligase(4CL),as the main biosynthetic enzyme of flavonoids and lignin,is closely related to the formation of plant flavonoids.In order to explore the relationship between the 4CL gene family of Glycyrrhiza uralensis and the synthesis and accumulation of isoglycyrrhizin,this study determined the content of isoglycyrrhizin in G. uralensis after drought stress,and the expression characteristics and bioinformatics analysis of the 4CL gene family of G. uralensis Fisch.to understand the relationship between the accumulation characteristics of isoglycyrrhizin and the 4CL gene of G.uralensis.The results showed that isoliquiritigenin was mainly accumulated in the roots of G.uralensis,and drought stress could significantly increase the content of isoliquiritigenin in the roots of G.uralensis.The content of isoliquiritigenin after 2 h of polyethylene glycol(PEG)stress was 3.91 times that of the control group(0 h).Drought stress could induce the up regulation of Gu4CL gene in the underground part.The expression levels of Gu4CL2, Gu4CL4 and Gu4CL5 were higher after PEG stress induction,while Gu4CL2 was most significantly up-regulated after PEG stress.The expression of Gu4CL2 was similar to that of isoliquiritigenin.Bioinformatics analysis of Gu4CL gene showed that 11 genes had two conserved polypeptide motifs Box Ⅰ(SSGTTGLPKGV)and Box Ⅱ(GEICIRG),and 11 genes were distributed on 11 Scaffold fragments.Promoter cis-acting element analysis showed that Gu4CL2, Gu4CL4 and Gu4CL5 genes contained more abscisic acid and jasmonic acid response elements than other Gu4CL genes.Therefore,drought stress may induce the synthesis of jasmonic acid and abscisic acid,regulate the expression of Gu4CL2, Gu4CL4 and Gu4CL5 genes in licorice roots,and increase the accumulation of isoliquiritigenin.This study provides a research basis for optimizing the cultivation techniques of G.uralensis,improving the adaptability and quality of cultivated land in saline-alkali land,exploring the role of Gu4CL gene family in G.uralensis,and exploring the synthesis mechanism of isoliquiritigenin.

Currently,traditional breeding methods cannot fully meet the demands of the cotton market and production. Therefore, molecular biotechnology can be employed to accelerate the development of new cotton varieties. Transcription factors have emerged as significant tools in the study of cotton gene function, genetics, and breeding during this period. Among them, the MYB transcription factor family is one of the largest transcription factor families present in many plants, and it performs multiple roles in plant growth and development. Hence, the MYB transcription factor holds high research value. While the function of MYB transcription factors has been extensively studied in model plants, research in non-model plants, particularly cotton, remains limited and is mostly concentrated in upland cotton. To further understand MYB transcription factors, this paper reviews the research progress on MYB transcription factors in various plants and cotton. It covers their classification basis, structural characteristics, evolution, roles in response to biotic and abiotic stress, cotton fiber development, and secondary metabolism. Additionally, it provides statistics on the known functions of cotton MYB transcription factors. This review aims to deepen our understanding of cotton MYB transcription factors and serve as an important reference for future studies on the functions and mechanisms of MYB transcription factors in different cotton species.

This study aims to provide theoretical guidance for the effective use of sister line materials,accelerate the promotion of the wheat Annong 1687 variety,and offer a reference for the genetic improvement of new wheat varieties.Wheat Annong 1687 and its sister lines and parents was used as materials.We combined phenotypic identification of field agronomic traits with genotypic identification of wheat materials using 55K microarrays.The data was analysed using SPSS and mapped it with the RIdeogram package.Significant differences in spike length were found between parents and some sister line combinations.Specifically,parents Annong 1106 and Xinong 822,and sister lines Annong 1687 and Xinong 822,showed very significant differences.The parental lines Annong 1106 and Xinong 822,as well as the sister lines Q6,8 and 105,133,exhibited a total of 909 differential SNP sites across 19 chromosomes.Notably,chromosome 5B contained 500 of these sites,which were concentrated in intervals of 63—87 Mb and 400—410 Mb.Cloning and sequencing of candidate genes within the intervals revealed that two genes encoding growth hormone-inducible proteins,TraesCS5B01G225000 and TraesCS5B01G058700,had missense mutations resulting in amino acid changes.Mutations in TraesCS5B01G225000 resulted in a codon change to the termination codon TGA and termination of amino acid synthesis.The bi-parental and sister line combinations had five and two missense mutations,respectively.It is hypothesized that these two genes may have an effect on the length of wheat spikes.

To study the genetic diversity of astringency in walnut germplasm resources,118 germplasm resources from 4 different astringency walnut populations were used as materials,which was studied by using SSR capillary electrophoresis fluorescence labeling technology,and the dendrogram of 4 populations was constructed.The results showed that a total of 93 allelic variants were detected by 12 pairs of primers,ranging from 2 to 18,with an average of 7.75 alleles detected by each pair of SSR primers.Polymorphism information content (PIC) ranged from 0.357 9 to 0.785 2,with an average of 0.541 1.The number of polymorphic sites (Np) was 91,the percentage of polymorphic sites (PPB) was 97.85%,the number of observed alleles (Na) was 1.978 5,the effective number of alleles (Ne) was 1.198 5,Shannon's information index (I) was 0.209 7,Nei's diversity index (H) was 0.126 3,number of total genetic diversity index (Ht) was 0.126 7 and intra-population genetic diversity index (Hs) was 0.122 2,indicating that the genetic diversity and variation of the four walnut populations were not high.The coefficient of genetic differentiation among populations was 0.035 5,indicating that the genetic variation within populations was 96.45%.The percentage of polymorphic loci in the slightly astringent population was the largest,which was 72.04%,indicating that the slightly astringent population had the richest genetic diversity among the four populations.UPGMA cluster analysis showed that the genetic identity between four populations ranged from 0.989 8 to 0.997 1,indicating that the genetic distance among four populations was close.The four populations can be divided into two groups at the coefficient of 0.993 4.The first group included unastringent,slightly astringent and more astringent populations,with the astringent populations as a separate group.By calculating the gene frequency of 93 locis in different populations,41 locis of 12 pairs of SSR primers were found to be able to distinguish the resources with different levels of astringency.SSR capillary electrophoresis fluorescence labeling technology was used to analyze the genetic diversity related to astringency in 4 walnut populations.According to the transcriptomic sequencing results,12 pairs of primers were selected for SSR analysis.A total of 93 SSR locis were amplified,and the percentage of polymorphic sites was 97.85%,indicating that the primers had high polymorphism and were suitable for genetic diversity analysis of walnut,which can be used for subsequent studies on genetic diversity and breeding of astringency in walnut.