Drought is one of the key factors limiting the yield of japonica rice.Exploring the genetic mechanism of drought tolerance in japonica rice at the germination stage,screening drought-tolerant candidate genes,and identifying their superior haplotypes provide a theoretical basis for deciphering the drought tolerance mechanism of japonica rice at the germination stage and developing related molecular markers.139 core germplasm resources of japonica rice from Northwest China were used as materials.Japonica rice seeds after radicle emergence were subjected to simulated drought stress using 15% PEG-6000.Six drought tolerance-related traits,including relative germination rate(RGR)and relative root length(RRL),were determined.Combined with 255 501 high-quality single nucleotide polymorphisms(SNPs),genome-wide association studies(GWAS)were conducted using the general linear model(GLM),mixed linear model(MLM),and fixed and random model circulating probability unification(FarmCPU)model.Haplotype analysis was further performed based on drought tolerance phenotypic data to identify potential candidate genes.The results showed that the mean values of the six drought tolerance traits followed a normal distribution,with coefficients of variation(CV)ranging from 3.43% to 56.25%.Among these traits,the relative root and shoot dry weight(RRSDW)exhibited the largest CV and showed a significantly positive correlation with relative root length(RRL),relative shoot length(RSL),and relative root and shoot fresh weight(RRSFW).GWAS for the six drought tolerance traits using high-density SNPs detected a total of 1 164 distinct SNPs,among which 379 were regarded as high-confidence SNPs loci and 6 as quantitative trait nucleotides(QTNs).By integrating the 50 kb upstream and downstream regions of these loci,14 significant SNPs loci were screened out,corresponding to 14 reliable quantitative trait loci(QTL)candidate intervals,which were mainly significantly associated with RRSDW.Genes within these 14 intervals were further predicted and analyzed using databases.With missense mutations in exons as the criterion,the gene LOC_Os03g56130,which encodes lichenase-2 precursor,was identified.Combined with drought tolerance phenotypes,haplotype analysis of this gene revealed that 5 missense mutations in exons constituted 4 haplotypes.Among them,Hap.2 showed a significantly higher RRSDW than the other haplotypes,being identified as the superior haplotype.These findings indicate that LOC_Os03g56130 is a potential candidate gene associated with drought tolerance in japonica rice at the germination stage.

The objective of this study was to identify novel major QTLs conferring plant height with breeding potential, and to screen favorable gene loci for marker-assisted selection, so as to provide theoretical basis and molecular resources for plant architecture improvement, lodging resistance improvement and high-yield breeding in wheat. A total of 162 doubled haploid (DH) lines derived from wheat cultivars AN1589 and LX6 were used as experimental materials, which exhibited significant phenotypic variation in plant height. The DH population was genotyped with the wheat 55K SNP array, and polymorphic molecular markers were screened to construct a high-density genetic linkage map. Combined with phenotypic data of plant height collected from six environments across three consecutive years, genome-wide QTL mapping was performed for plant height traits. Four environmentally stable major QTLs were detected via inclusive composite interval mapping (ICIM), which were distributed on chromosomes 4B, 4D, 5D and 6D, individually explaining 7.08%-39.31% of the phenotypic variation. Among them, Qph.ahau-4B was steadily identified in all six environments and BLUP analysis, explaining 28.65%-39.31% of the phenotypic variation. This locus covered a physical interval of 20.61-22.26 Mb, flanked by markers AX-109018264 and AX-109968140, with an additive effect ranging from -9.34 cm to -8.68 cm. This QTL presented prominent genetic effect and high environmental stability, and was verified as a novel and previously unreported major dwarfing QTL by literature comparison. In conclusion, a novel major and environmentally stable QTL for plant height was successfully identified on chromosome 4B. It lays a foundation for subsequent fine mapping, gene cloning and molecular marker-assisted breeding in wheat.

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

This study aimed to clone the wheat leaf rust resistance-related gene TaRLKS.2 and detect the kinase activity of its expressed protein,in order to lay a foundation for further exploring the mechanism of TaRLKS.2 in wheat resistance against Puccinia triticina infection and to provide new genetic resources for wheat leaf rust resistance breeding.Using cDNA from the leaf rust-resistant near-isogenic line TcLr26 as a template,the full-length coding region of TaRLKS.2 was cloned.The domain structure of the TaRLKS.2 protein was analyzed using GSDS 2.0 software.A pET28a-TaRLKS.2 prokaryotic expression vector was constructed,and the TaRLKS.2-His fusion protein was induced with IPTG,followed by purification using Ni-NTA affinity chromatography.The kinase activity of the expressed TaRLKS.2 protein was detected using the phosphorylation substrate MBP and Western Blotting.The results showed that the coding region of TaRLKS.2 was 2 244 bp in length,encoding a protein containing two kinase domains with conserved kinase active centers and ATP-binding sites,exhibiting typical characteristics of the RLCK family.Under conditions of 28 ℃ and 0.25 mmol/L IPTG,the TaRLKS.2-His recombinant protein was induced to form soluble protein.After adding the substrate MBP,ATP,phosphate buffer,and protease inhibitors to the purified recombinant protein,the reaction was carried out at 30 ℃ for 30 min.Detection with an anti-phospho-Ser/Thr-Pro antibody confirmed that TaRLKS.2 possesses protein kinase activity.It is speculated that TaRLKS.2 may regulate wheat immune pathways by phosphorylating downstream signaling components,thereby enhancing wheat resistance to leaf rust.

In order to identify the pathogenic source of Tomato chlorosis virus disease and clarify the taxonomic status of Tomato chlorosis virus (ToCV)in the Shijiazhuang area of Hebei Province,an analysis of the phylogenetic relationships of diseased samples collected from the Tomato Planting Base in Dahe Town,Shijiazhuang,was conducted.Additionally,an efficient method for ToCV identification was established.The published ToCV sequences in NCBI database were selected for similarity analysis,and the full-length RNA1 and RNA2 sequences of ToCV,Heat-shock protein 70 homologue(Hsp70h)and coat protein(CP)sequences were used to construct the phylogenetic tree.The results showed that the nucleotide(nt)sequences of RNA1 and RNA2 of the ToCV-SJZ shared 85.2% and 92.4% similarity,respectively,with the corresponding sequences of the ToCV-XS Taiwan strain,indicating the closest genetic relationship.The evolutionary analysis showed that ToCV could be divided into two clades,one represented by the Florida strain and the other by the Taiwan strain.According to the CP gene sequences of ToCV-SJZ,ToCV-XS and ToCV-Florida strains,a RT-qPCR assay system based on SYBR Green Ⅰ was established.In conclusion,this study clarifies the taxonomic status of the ToCV-SJZ isolate and provides technical support for the identification of disease-resistant germplasm resources and scientific prevention and control.

Serine hydroxy methyl transferase(SHMT) is a conserved pyridoxal phosphate-dependent enzyme that catalyzes the reversible conversion between serine and glycine. It is also an essential enzyme for nucleotide biosynthesis, folate, and amino acid metabolism, playing a critical role in plant development and enhancing the ability of crops to resist stress.The watermelon inbred line TN07011 was used as the material.ClSHM78 was cloned by RT-PCR technology and subjected to bioinformatics analysis.An overexpression vector was constructed and transformed into Arabidopsis thaliana. The T₃ transformed lines were subjected to low-temperature and salt stress treatments,by determining phenotypic and physiological indices, the function of ClSHMT8 in abiotic stress was explored.The results showed that ClSHMT8 was 1 749 bp in full length, encoding 582 amino acids. NCBI-CDD conserved domain prediction indicated that the protein contained a typical SHMT domain.Phylogenetic tree analysis showed that this protein shares a close genetic relationship with its homologous proteins from cucumber and bitter melon.Heterologous expression of ClSHMT8 in Arabidopsis thaliana yielded three T₃ lines, which exhibited significantly larger rosette leaf area, earlier flowering period, and larger seed size compared to the wild type. Under low-temperature and salt stress, the T₃ plants showed significantly higher contents of soluble protein(SP) and chlorophyll(Chl), as well as activities of antioxidant enzymes such as peroxidase(POD), superoxide dismutase(SOD), and catalase(CAT), while the content of malondialdehyde(MDA) was significantly lower than that of wild type. Taken together, ClSHMT8 not only positively regulates plant growth and development, but also enhances plant tolerance to abiotic stresses.These results demonstrate that ClSHMT8 plays an important role in the response of Arabidopsis thaliana to abiotic stress environments.

In order to systematically analyze the sequence characteristics and expression pattern of BolMYC2 in broccoli,and to explore the role of BolMYC2 in response to the infection of soft rot pathogens in broccoli,the coding sequence of BolMYC2 gene was cloned by PCR amplification using broccoli cDNA as a template.The basic properties of BolMYC2 protein were analyzed by bioinformatics software.BolMYC2-EGFP fusion expression vector was constructed and transiently transformed into Nicotiana benthamiana to analyze the subcellular localization of BolMYC2 protein.The transcriptional expression characteristics of BolMYC2 in different tissues of broccoli and the transcriptional expression characteristics of BolMYC2 under the stress of soft rot pathogens were analyzed by qRT-PCR.The results showed that the coding sequence of BolMYC2 was 1 833 bp,encoding 610 amino acids.The theoretical molecular weight was 66.297 ku and the theoretical isoelectric point was 5.21.BolMYC2 was an unstable hydrophilic protein without transmembrane structure and signal peptide.Conserved domain analysis showed that BolMYC2 protein contained the conserved domains bHLH-MYC_N and bHLH_SF of MYC transcription factor family.Phylogenetic tree analysis showed that BolMYC2 protein was closely related to brussels sprouts.Analysis of the cis-acting elements of the BolMYC2 promoter showed that it contained cis-acting elements involved in light response,hormone response,stress response,and multiple MYB,MYC,and WRKY transcription factor recognition elements.Subcellular localization results showed that BolMYC2 was localized in the nucleus.BolMYC2 was highly expressed in leaves and seeds.In addition,the expression level of BolMYC2 decreased first and then increased after broccoli was infected by soft rot pathogens,suggesting that BolMYC2 plays an important role in broccoli response to biotic stress.

Small heat shock proteins(sHSP)play an important regulatory role as molecular chaperones in plant stress resistance.To investigate the biological function of sHSP in pepper,the heat-tolerant pepper material 17CL30 was used as the experimental material.The sHSP-encoding gene CaHSP21 was cloned and subjected to bioinformatics analysis.Its expression patterns in different tissues and under high-temperature stress were analyzed by Quantitative Real-time PCR.The results showed that the full-length CDS of CaHSP21 was 705 bp,encoding 234 amino acids.Physicochemical analysis revealed that the molecular mass and theoretical isoelectric point(pI)of CaHSP21 were 25.74 ku and 6.53,respectively,indicating it is a neutral protein.Its instability index was 37.39,it is classified as a stable protein.CaHSP21 contained the typical HSP20 domain of the sHSP family,and its secondary structure was primarily composed of random coils.Protein phylogenetic analysis showed that CaHSP21 had the closest genetic distance to Capsicum chinense and Capsicum baccatum,and they belonged to the same evolutionary branch.Tissue-specific expression analysis indicated that CaHSP21 expression was lowest in flowers and highest in fruits.Under high-temperature stress,CaHSP21 was induced by high temperature,and its relative expression level peaked after 3 hours of treatment.Subcellular localization results demonstrated that CaHSP21 was localized in the plasma membrane and cell nucleus.In conclusion,CaHSP21 is a key gene involved in pepper's response to high-temperature stress.

The chloroplast genome of Cucurbita ficifolia was assembled,annotated and analyzed to reveal the phylogenetic relationship between C.ficifolia and other Cucurbitaceae plants.The Illumina NovaSeq 6000 platform was used to sequence C.ficifolia,and its sequence characteristics,gene type,gene repeat sequence,codon preference and species evolution were analyzed.The chloroplast genome of C.ficifolia was 157 631 bp in length with a GC content of 37.15%.A total of 141 genes were annotated,comprising 60.99% protein-coding genes,33.33% tRNA genes,and 5.67% rRNA genes.These genes were categorized into four groups:photosynthesis-related genes,self-replication genes,other genes,and genes of unknown function.Simple sequence repeat analysis was performed on the chloroplasts of C.ficifolia,and a total of 192 SSR loci were detected.Codon preference analysis showed that there were 31 codons with RSCU values greater than 1.00,and 29 codons ended with A or U.The boundary genes of C.ficifolia were different from those of 8 other Cucurbitaceae species,and the expansion and contraction phenomena were obvious in the IR region and LSC region.The results of evolutionary analysis showed that C.ficifolia was in the same branch with C.moschata,C.maxima,C.pepo,C.argyrosperma var. palmeri,C.foetidissima,C.okeechobeensis subsp. martinezii and C.lundelliana.C.ficifolia was distant in relationship with other species of Cucurbitaceae.The chloroplast genome of C.ficifolia was assembled,and annotated with 141 genes and 192 SSR loci.Its evolutionary relationship is relatively distant with the selected Cucurbitaceae species.This study provides a theoretical basis for phyletic evolution and molecular breeding strategies in C.ficifolia.

The molecular chaperone heat shock protein HSP40 binds to HSP70 through the J domain to assist in protein folding and participates in various stress responses in plants.However,the mechanism and function of NtHSP40 gene family in response to UV-B radiation remain unknown.Through bioinformatics methods,the members of the NtHSP40 gene family were identified at the whole-genome level,and phylogenetic analysis,collinearity analysis,cis-acting elements of the promoter,chromosomal localization,gene structure analysis and physical and chemical property analysis were carried out to clarify the basic characteristics of the NtHSP40 gene family.In tobacco seedlings treated with UV-B radiation,RNA-sequencing analysis was performed to investigate the eight differentially regulated genes according to the transcriptome data.The expression levels of candidate genes belonging to NtHSP40 gene family were investigated by Quantitative Real-time PCR(qRT-PCR)experiments and the total polyphenol content of metabolites was quantified by spectrometry.The results showed that there were 48 NtHSP40 genes in the genome of tobacco K326.These genes were distributed on 20 chromosomes and all encoded J domain proteins which can be further divided into three subfamilies.The variable gene structure and protein sequence characteristics indicated the diversification of the functions of the NtHSP40 gene family members.Among them,eight NtHSP40 genes responded to UV-B radiation,and was correlated with the total polyphenol content in tobacco.The research findings establish a foundation for further elucidating the functions of the NtHSP40 gene family in plant stress responses and secondary metabolism in tobacco,and for enhancing the quality of tobacco leaves.

To further investigate the function of cotton basic helix-loop-helix(bHLH)transcription factors in salt stress response,we introduced the cotton bHLH transcription factor GhbHLH149-like(previously identified by our team)into wild-type Arabidopsis thaliana via Agrobacterium-mediated transformation,and verified the function of GhbHLH149-like transcription factor in Arabidopsis.Protein structure prediction indicated that the secondary structure of GhbHLH149-like protein is mainly composed of α-helice and random coil,and it was a non-transmembrane hydrophilic protein.Quantitative Real-time PCR(qRT-PCR)results showed that the expression level of the GhbHLH149-like gene in transgenic Arabidopsis was significantly higher than wild-type.The results of phenotypic and physiological index assays indicated that the seed germination rate and root length of transgenic seedlings were significantly higher under salt stress,and the growth performance of transgenic seedlings was markedly better.Meanwhile,the contents of hydrogen peroxide(H₂O₂)and superoxide anion(O₂^-)were significantly lower,whereas the superoxide dismutase(SOD)activity was significantly higher in the transgenic lines.Taken together,GhbHLH149-like gene can significantly enhance the salt tolerance of Arabidopsis,presumably by participating in reactive oxygen species(ROS)scavenging.This study lays a theoretical foundation for the in-depth dissection of the molecular mechanism of cotton GhbHLH149-like gene in salt stress response.

In order to determine the genetic law of the first flower node in kenaf,an F₂ population consisting of 148 individual plants was developed from the inbred lines K215 and K89,which showed significant differences in the first flower node position.The high-density genetic linkage map of kenaf was constructed using RAD-seq for bin marker genotyping.The map contained 3 418 bin markers unevenly distributed on 18 chromosomes,with a total genetic distance of 864.61 cM and an average genetic distance between markers of 0.25 cM.Based on the phenotypic data and genetic map of the first flower node,the first flower node QTL named FFN6 was detected using an interval mapping method.This QTL was located on chromosome 6,with a LOD value of 8.66,a phenotypic contribution rate of 13.77%,and an additive effect value of -19.97.The results indicated that the allele locus comes from the paternal K215 and had a negative regulatory effect on the first flower node.Based on the gene annotation information within the QTL mapping interval,seven candidate genes were preliminarily screened and obtained.The qRT-PCR results showed that under short-day conditions,the expression levels of Hc-bHLH6 and Hc-MYB6 genes significantly increased.The function of the two genes promotes the flowering of kenaf plants,which may be closely related to the node of the first flower of kenaf plants.

AINTEGUMENTA-LIKE 5(AIL5)is one of the key genes regulating plant growth and development during somatic embryogenesis.To explore the function of the AbAIL5 gene in the somatic embryogenesis of Amorphophallus bulbifer and to accelerate its molecular breeding process,the AbAIL5 gene was cloned from the bulbils of A.bulbifer,and its nucleotide sequence and protein structure were analyzed using bioinformatics tools.The open reading frame(ORF)of AbAIL5 was 654 bp in length,encoding 217 amino acids.Bioinformatics analysis indicated that the AbAIL5 protein was a hydrophilic protein containing a conserved AP2 superfamily domain,without transmembrane domains or signal peptides.Phylogenetic analysis showed that AbAIL5 was most closely related to that of Platanthera zijinensis.Subcellular localization analysis revealed that the protein resides specifically within the cell nucleus.Regarding expression patterns,qRT-PCR analysis revealed that AbAIL5 was expressed in the aerial parts(leaves,petioles,and bulbils)of A.bulbifer,with higher expression levels observed in the early stages of bulbil development.The relative expression levels in petioles,leaves,and bulbils were 1.00,0.55,and 1.42,respectively.For functional verification,an AbAIL5 overexpression vector was constructed and transformed into A.bulbifer callus mediated by Agrobacterium tumefaciens.After 30 days of culture,the AbAIL5 overexpression group showed significantly increased growth rates and germination numbers,as well as an improved overall survival rate compared with the control group.At 70 days,plant growth in the overexpression group was significantly superior to that of the control.qRT-PCR results confirmed that the expression level in the overexpression group was 5.34-fold higher than that of the control.These results indicated that AbAIL5 facilitates adventitious bud differentiation and callus proliferation,providing a theoretical basis for revealing the function of AbAIL5 in the development of A.bulbifer and for improving its genetic transformation efficiency.

The large-scale production of soybean hybrids and the utilization of hybrid vigor are key to germplasm resource innovation and variety improvement.By mining and utilizing soybean male sterility genes,more advantageous soybean varieties can be cultivated,thereby improving soybean yield and quality.This study selected the male sterile mutant population that appeared in the hybrid offspring of Baoquanling green soybean(female parent)and Heinong 44(male parent)as the experimental material.Flower buds were selected from soybean fertile and sterile plants,the anthers and pollen cytology were observed,and a high-throughput sequencing platform was used to perform transcriptome sequencing on the flower and anther tissues of soybean fertile and sterile plants.Annotated,screened and analyzed the sequencing results.The results indicated that abnormal development of the tapetum layer and high pollen sterility in sterile plants were important causes of male infertility.The pentose and glucuronate interconversion pathway(ko00040:Pentose and glucuronate interconversion)was a key pathway affecting soybean fertility,and the genes encoding pectin methylesterase(PEM)and pectin lyase(PL)were downregulated in expression.The differentially expressed gene protein interaction network indicated that the pectin lyase gene was a key gene affecting the interconversion pathway between pentose and glucuronic acid.The determination of pectin lyase content showed that the pectin lyase content in sterile plant anthers was significantly reduced.The qRT-PCR detection results were consistent with the RNA-Seq sequencing results.It is speculated that the downregulation of these genes may cause a decrease in pectin lyase activity,thereby affecting the development of the tapetum and leading to male infertility.

In order to clarify the function of the protein phosphatase 2C gene GmPP2C72 in salt stress response and its salt-tolerance molecular mechanism,bioinformatics analysis of GmPP2C72 protein was conducted, GmPP2C72 gene was cloned to construct the overexpression vector,and then was transferred into Lotus japonicus using the cotyledon node genetic transformation method mediated by Agrobacterium tumefaciens.Finally,the phenotype,physiological indicators and the expression levels of salt tolerance related genes of transgenic plants were analyzed under salt stress condition.The results showed that GmPP2C72 gene contained an open reading frame of 1 206 bp,encoding 401 amino acids,and the encoded protein belonged to the member of the class A subfamily of PP2C family.GmPP2C72 gene did not exhibit tissue expression specificity,it was expressed in various tissues of soybean,and the expression of this gene was induced by salt stress.The overexpression vector of GmPP2C72 gene was successfully constructed.GmPP2C72 gene was successfully transferred into Lotus japonicus,and three transgenic lines with GmPP2C72 gene were successfully obtained.Compared with wild type,the three transgenic lines maintained better growth state,with significantly higher contents of chlorophyll and proline and lower relative plasma membrane permeability and malondialdehyde content in leaves of Lotus japonicus under salt stress.In addition,the expression levels of four key salt tolerance related genes,LjLEA, LjCOR,LjRD29B,and LjP5CS were significantly higher compared with wild type.Overall,the overexpression of GmPP2C72 gene enhances the salt tolerance of Lotus japonicus by reducing the degree of membrane lipid peroxidation and the permeability of the plasma membrane,increasing the contents of chlorophyll and proline in the leaves,and enhancing the expression levels of salt tolerance related genes.

Discovery and mapping of disease-resistant genes in wheat varieties can provide resistance sources,genes and markers for the breeding of new wheat cultivars.KU3067/Apav BC₁F₅ population were used to evaluate for leaf rust resistance at seedling and adult plant stage in Baoding,Hebei Province,and leaf rust resistance gene(s)were mapped using molecular marker technology.At the seedling stage,KU3067 was highly susceptible to three leaf rust physiological races while Apav was highly resistant.The 148 BC₁F₅ lines showed a separation ratio of one pair of genes,indicating that there was a dominant gene in Apav.It was confirmed that the gene was the known leaf rust resistance gene Lr13 using molecular marker technology.Both parents showed high resistance at the adult stage,and the population showed continuous distribution,which was consistent with the characteristics of quantitative traits.Combined with the DArT-Seq data of the population,five leaf rust resistance QTLs were mapped in this population.Lr13,from Apav,was detected in two environments and explained 51.60% and 25.08% of the phenotypic variance,respectively.Lr67 from KU3067 was detected in two environments,explaining 4.98%—7.30% of the phenotypic variance,respectively.QLr.hebau-2AS,from Apav,explained 6.68% of the phenotypic variance.QLr.hebau-5DL.1 from Apav,and QLr.hebau-5DL.2 from KU3067 explained 23.74% and 8.41% of the phenotypic variance,respectively,the two gene might be novel.This study clarified the resistance performance characteristics of the KU3067/Apav population in the Baoding environment,mapped multiple genetic loci including Lr13,Lr67,and three potentially novel leaf rust resistance QTLs,and developed closely linked molecular markers.This provides valuable genetic resources and markers for wheat leaf rust resistance breeding in China.

The abscisic acid(ABA)receptor protein PYL,as the initial component of ABA signaling transduction,actively participated in ABA-mediated signaling transduction in plants and played a crucial role in regulating plant resistance to stress.In our previous study,based on transcriptome data,the gene Cla97C07G133100.1,which was negatively responsive to salt stress in watermelon,was screened out and named ClPYL8.To further explore the biological function of the watermelon ABA receptor ClPYL8,using watermelon TN07011 as the experimental material,the structure and function of ClPYL8 were analyzed using bioinformatics,subcellular localization,and RT-qPCR methods.The results showed that the open reading frame of ClPYL8 gene was 702 bp,encoding 233 amino acids.Protein conserved domain prediction revealed that ClPYL8 contained the PYR/PYL/RCAR-like conserved domain,belonging to the SPRBCC superfamily.The promoter cis-acting element analysis indicated that the ClPYL8 promoter contained various elements responsive to stress,auxin,and light.The seamless cloning technique was used to construct the EGFP fusion expression vector,which was then transformed into tobacco.The results showed that ClPYL8 was localized in the nucleus and cell membrane.Tissue-specific(RT-qPCR)analysis revealed that the gene exhibited the highest expression level in seeds and the lowest in fruits and female flowers;under salt stress,the expression of ClPYL8 showed a downregulation trend;under drought stress,the gene expression first decreased and then increased;under low-temperature stress,the gene expression first increased and then decreased.In conclusion,ClPYL8 actively responded to abiotic stress and might regulate watermelon's stress response through the ABA signaling pathway,suggesting a potential negative regulatory role in salt stress and positive regulatory roles in drought and low-temperature stress.

Jasmonates play a crucial role in regulating fruit ripening and quality formation.The JAZ proteins are key repressors of the jasmonic acid signaling pathway.To investigate the function of PavJAZ1 in sweet cherry fruit ripening and quality regulation,the PavJAZ1 gene was identified and cloned from the sweet cherry genome.Bioinformatics analysis was performed to explore its physicochemical properties,protein structure,conserved domains,and cis-acting elements in the promoter region.The gene expression pattern of PavJAZ1 in fruits was analyzed by Quantitative Real-time PCR.The subcellular localization of the PavJAZ1 protein was observed,and the interaction between PavJAZ1 and PavMYC2 was verified using the bimolecular fluorescence complementation(BiFC).The results of gene identification and cloning showed that the open reading frame of the PavJAZ1 gene was 837 bp in length,encoding 278 amino acids.The molecular weight of the PavJAZ1 was 30.09 ku,the theoretical isoelectric point was 9.21,the instability index was 54.14,and the grand average of hydropathicity was -0.564,indicating that it was an alkaline hydrophilic and unstable properties.Protein structure analysis revealed that PavJAZ1 was mainly composed of random coils and α-helices,containing two conserved domains,the Tify domain and the Jas domain.It also exhibited high homology with proteins from other species,among which it showed the highest homology with that of peach,a plant that also belonged to the genus Prunus of the Rosaceae family.Subcellular localization analysis showed that the PavJAZ1 protein was localized in the nucleus.The expression level of PavJAZ1 increased significantly in the early stages of fruit development and then gradually decreased.This result suggested that PavJAZ1 may play different roles at different developmental stages.In addition,the BiFC results showed that PavJAZ1 interacted with PavMYC2,confirming that PavJAZ1 participates in the jasmonic acid signaling pathway.

The Kip-related protein(KRP)gene family acts as a key regulator of the cell cycle,modulating cell division and endoreduplication by inhibiting cyclin-dependent kinase(CDK)activity,and participates in plant abiotic stress responses.To investigate the structural characteristics and environmental stress response profile of PbKRP2 in Pyrus betulifolia,the gene was cloned using reverse transcription PCR(RT-PCR).Bioinformatic analysis was conducted to elucidate its sequence features,predicted protein interactions,and phylogenetic relationships.Quantitative Real-time PCR(qRT-PCR)was employed to assess the tissue-specific expression characteristics of PbKRP2 and its expression patterns under various stress conditions.Results showed that PbKRP2 was 585 bp in length,contained 4 exons and 3 introns,and encoded a 194-amino acid protein.The predicted protein had 25 potential phosphorylation sites,a O-glycosylation site,and a conserved C-terminal cyclin-dependent kinase inhibitor(CDI)domain.Phylogenetic analysis indicated that PbKRP2 was closely related to PyKRP2 in Prunus yedoensis var.nudiflora,PpKRP2 in Prunus persica,and MdKRP2 in Malus domestica.Protein interaction network screening identified potential interactions between PbKRP2 and 10 proteins.Analysis of the static structure of potential interacting proteins suggested that PbKRP2 formed spatial interaction conformations with its target proteins PbCDKB1;1 and PbCYCD7;1,respectively.The promoter region of PbKRP2 contained multiple cis-acting regulatory elements responsive to environmental factors and hormonal signals.qRT-PCR analysis demonstrated that PbKRP2 transcript levels were significantly higher in stems and leaves compared to roots,petals,and fruits.Furthermore,PbKRP2 transcription levels were consistently upregulated under low-temperature stress and abscisic acid(ABA)treatment,but significantly downregulated under dehydration and salt stress.In summary,these findings provide a candidate gene for stress-resistant breeding of pear and genetic improvement of fruit trees by elucidating the sequence characteristics of PbKRP2 and its differential response patterns to abiotic stresses.

Evaluating the pedicel length trait of cucumber germplasm resources and exploring the related variation sites can help improve the breeding efficiency of cucumber quality.This study first evaluates the fruit pedicle length traits of 229 cucumber germplasm resources on the day of flowering and during the commercial fruiting period.Then,using the identified medium to long pedicle self-cross lines 9D6 and 6457 as parents,and short pedicle self-cross lines MT and 6429 as parents,F₂ populations were constructed.QTL-seq was used to locate the QTLs controlling cucumber pedicel length traits,and Real-time Quantitative PCR(qRT-PCR)was employed to analyze the expression patterns of candidate genes and validate variant sites.The results showed that the pedicel length on the day of flowering and during the commercial melon period was positively correlated with the germplasm resources,and the variation of the pedicel length on the day of flowering and during the commercial melon period was 0.4—6.2 cm,0.4—5.3 cm,respectively.Genetic analysis showed that the broad genetic forces of cucumber pedicel length were 69.4%,94.4%,respectively,showing typical quantitative trait inheritance characteristics.Six and five QTL loci related to pedicel length on the day of flowering were detected in two sets of F₂ population,respectively.Two QTL intervals 0.840—7.350 Mb on chromosome 5 and 17.735—31.087 Mb on chromosome 6 were co-localized in the two sets of populations.CsPG1(CsaV3_5G010970)and CsROS1 (CsaV3_6G040410)were predicted within the QTL interval of co-location.A non-synonymous SNP311 C/T was found at the conserved domain of CsPG1 coding sequence(CDS)in the medium-long fruit pedicel parental lines 9D6 and 6457.In addition,there was a 9-bp base insertion in the CsPG1 promoter of 9D6,which contained TATA-box,which may be related to its significantly higher expression than other parents on the day of flowering.A 3-bp deletion was found at the CDS of CsROS1 in the short fruit pedicel parental lines 6429,resulting in the loss of one serine residue in the encoded protein.A synonymous SNP3282 T/C was detected at the CDS in the long fruit pedicel parental lines 9D6 and 6457,causing the preferred codon UAU to change to the non-preferred codon UAC.CsPG1 and CsROS1 were considered to be key candidate genes for cucumber fruit pedicel length.

Research on the expression pattern and function of FaesANT gene in long-homostyle(LH)common buckwheat and uncovering its roles involved in floral organ development will help to provide gene resources for the molecular breeding and germplasm innovation of buckwheat.The full-length FaesANT gene was isolated from long-homostyle common buckwheat by combining PacBio SMRT(Single Molecule Real-Time)sequencing and RACE technique.FaesANT expression levels in different tissues and floral buds at sequential developmental stages of long-homostyle common buckwheat were separately detected by using paraffin sections and qRT-PCR.Moreover,the FaesANT gene function involved in floral development was investigated by using virus-induced gene silencing(VIGS)in long-homostyle common buckwheat.The results were as follows,FaesANT cDNA(1 982 bp)comprised a 1 632 bp ORF encoding a 543 amino acid transcription factor.Phylogenetic tree analysis grouped the FaesANT into the eudicots ANT lineage.FaesANT expression levels were mainly detected in stamen,pistil and young fruit of long-homostyle common buckwheat and showed no significant differences among them.However,only weak transcripts were detected in the stems,leaves and tepals.During floral bud differentiation,obvious expression of FaesANT gene was detected when pistil and stamen primordia formation,and FaesANT expression level reached peak during filament elongation stage of stamen.Then,FaesANT expression decreased gradually with the floral bud development and dropped to the lowest level after flowering.TRV2-FaesANT-treated long-homostyle plants with strong phenotypic changes produced flowers with style-and filament-length decreased,and stamen number reduced.In addition,some TRV2-FaesANT-treated long-homostyle plants displayed part stamens with anther homeotically converted into tepaloid structure.Moreover,FaesANT expression decreased significantly in the flowers of FaesANT-silenced long-homostyle plants with strong phenotypic changes.It suggested that FaesANT was mainly involved in regulating filament and style length,and determining stamen number of common buckwheat based on the gene expression pattern and phenotypes of VIGS-treated long-homostyle common buckwheat flowers.

Cinnamic acid 4-hydroxylase (C4H),a key enzyme in the flavonoid synthesis pathway,plays a crucial role in plant stress resistance.To clarify the bioinformatic functions of the C4H gene family in three medicinal liquorice species (Glycyrrhiza uralensis Fisch.,Glycyrrhiza inflata Bat.,and Glycyrrhiza glabra L.),investigate their expression characteristics under abiotic stress,as well as their relationship with the synthesis and accumulation of liquiritin,we used bioinformatics methods to analyze the C4H gene family members,and combined transcriptome data and RT-qPCR validation to understand their expression under abiotic stress and their relationship with liquiritin content.Results showed that a total of 11 C4H genes were identified in the whole genomes of the three medicinal liquorice species and were named GuC4H1—3,GiC4H1—3,and GgC4H1—5,respectively.These genes were distributed on 2 chromosomes in G.uralensis and G.inflata and on 3 chromosomes in G.glabra.Although the gene members differed in physicochemical properties such as the number of amino acids and isoelectric points,all of them were predicted to be localized in the endoplasmic reticulum and contain a C-terminal heme-binding domain (PFGVGRRSCPG).Expression pattern analysis indicated that the 11 C4H genes were expressed in different tissues of the three medicinal liquorice species,and abiotic stress significantly induced the upregulation of C4H gene expression in the underground parts.Among them,the expression of GgC4H5 was the highest under salt stress for 24 h,which was 1.68 times that of the control group (24 h);the expression of GuC4H3 was the highest under drought stress for 2 h,which was 3.03 times that of the control group (2 h).In addition,moderate abiotic stress significantly increased the liquiritin content in the underground parts of the three liquorice species.Under NaCl stress for 2 h,the increase in liquiritin content in G.inflata was the most significant,which was 2.92 times that of the control group (2 h);under PEG stress for 24 h,the increase in liquiritin content in G.glabra was the most significant,which was 2.31 times that of the control group (24 h).GgC4H5 showed a similar expression trend to liquiritin content under salt stress,while GuC4H2,GuC4H3,GiC4H2,and GiC4H3 showed a similar expression trend to liquiritin content under drought stress.Therefore,these genes are likely to be the key genes involved in the response to abiotic stress and liquiritin synthesis in the underground parts of the three medicinal liquorice species.

SAUR genes play an important regulatory role in plant growth and development.The plant hormones ABA and GA₃ play key roles in the plant growth-stress response.To investigate whether the rice OsSAUR23 gene is regulated by ABA and GA₃,it analyzed the cis-regulatory elements in the promoter region of the OsSAUR23 gene.Using transgenic Arabidopsis lines expressing OsSAUR23 and wild-type Arabidopsis as experimental materials,the plants were treated with exogenous hormones.By measuring agronomic traits,physiological indicators,and the expression of genes related to acid growth and the ABA pathway,the study elucidated the mechanism by which the rice OsSAUR23 gene responds to ABA and GA₃.The results indicated that the promoter region of the OsSAUR23 gene contained cis-acting elements for ABA and GA₃,and that the expression of the OsSAUR23 gene in rice significantly increased under ABA and GA₃ treatment.Regarding agronomic traits,as ABA concentration increased,the germination rate,root length,plant height,and fresh weight of transgenic Arabidopsis were inhibited with increasing ABA concentrations,but plant height and fresh weight remained significantly higher than those of WT.GA₃ treatment induced a bidirectional regulatory pattern in transgenic Arabidopsis,low concentrations promoted germination rate and root length,while high concentrations suppressed them.Plant height and fresh weight of transgenic lines were consistently higher than WT under GA₃ treatment.Physiologically,SOD activity in transgenic Arabidopsis increased significantly with ABA concentration,while POD activity,CAT activity,and MDA content showed no significant difference compared to WT.Soluble protein content was significantly higher in transgenic lines.Under GA₃ treatment,SOD and POD activities increased,CAT activity initially rose and then declined,MDA content first decreased and then increased,and soluble protein content remained elevated compared to WT.Furthermore,both ABA and GA₃ treatments promoted a decrease in pH in the roots and other tissues of the overexpression lines,although this effect was inhibited under high hormone concentrations.qRT-PCR results demonstrated that overexpression of OsSAUR23 significantly downregulated the expression of AtPP2C.D family genes (AtAPD5,AtAPD6,and AtAPD9),which lifted the inhibition of plasma membrane H⁺-ATPase activity,promoted proton efflux,and ultimately led to acidification in root zones and other regions.Meanwhile,overexpression of OsSAUR23 also caused differential changes in the expression of AtPP2C.A members,suggesting its potential role in modulating ABA signaling transduction.OsSAUR23 positively regulates plant growth in response to ABA and GA₃.

Based on quantitative trait locus(QTL)mapping of grain weight and grain shape-related traits in a backcross inbred line(BIL)population derived from the cross between indica rice variety Changhui 891 and japonica rice variety 02428,this study provided valuable genetic resources for elucidating the genetic basis of grain development and molecular breeding for high yield and quality in rice.A high-density genetic map was used to conduct QTL analysis for thousand-grain weight,grain length,grain width,grain thickness,and length-width ratio under two environments.Transcriptome data of developing grains at 5,10,and 15 days after flowering from both parents were integrated to analyze candidate genes within QTL regions.Frequency distribution indicated that grain weight and shape traits in the BIL population exhibited typical quantitative characteristics,confirming the suitability for QTL mapping.A total of 37 QTLs were detected across both environments,including 18 specific to Hainan(Sanya),12 specific to Jiangxi(Nanchang),and 7 consistently identified in both locations.These QTLs were distributed on chromosomes 1—5,7,8,11,and 12,comprising 7 for thousand grains weight,5 for grain length,7 for grain width,11 for length-width ratio,and 7 for grain thickness.The analysis showed that the LOD values of these QTLs ranged from 2.79 to 43.10,with a contribution rate of 1.36% to 46.12%.By integrating differentially expressed genes from developing grains of both parents at three stages,28 candidate genes were prioritized from the 37 QTL regions.Among these,LOC_Os11g10480 and LOC_Os11g10100 were identified as key candidates for further functional validation.

The screening of aromatic rice germplasm resources and the rapid identification and utilization of aromatic genes have practical significance for the genetic breeding of aromatic rice.In order to clarify the aroma types of aromatic rice materials in Heilongjiang Province and improve the breeding efficiency of aromatic rice varieties,180 aromatic rice varieties approved and promoted in Heilongjiang Province were sequenced.Compared with the reference genome Nipponbare,the sequencing results were divided into two types of mutations.The first type was the 8 bp deletion of exon 7 and three base substitutions(Badh2-E7 type),including 177 varieties.The second type was the 7 bp deletion of exon 2(Badh2-E2 type),which contained only three varieties.According to these two mutation types,two specific KASP molecular markers were designed to accurately detect the aroma types of E7 and E2 in rice.Through the identification of 180 aromatic rice varieties,the typing results were consistent with the sequencing results.Two hybrid populations were constructed by using E7 aromatic rice variety Wuyoudao 4 and E2 aromatic rice variety Songkejing 134 with non-aromatic japonica rice varieties Weinong 101 and Dongfu 114,respectively.The F₂ generation of the hybrid population was screened and identified for aroma genes,and the identification results were consistent with 100%,which verified the accuracy of KASP molecular markers.The 620 japonica rice parents created and collected by our laboratory were further identified by KASP markers,and 248 aromatic rice materials were screened.It was found that these parents were all Badh2-E7 type genes.At the same time,the existing 1 220 F₇ generation materials with E7 aromatic rice as parents were genotyped,including 293 aromatic rice materials,906 non-aromatic rice materials and 21 heterozygous rice materials.Five aromatic and non-aromatic rice materials were randomly selected for sequencing verification.The sequencing results verified that the KASP molecular marker typing was correct.This study provided new markers and materials for breeding new fragrant rice varieties in cold regions by using molecular marker assisted selection.

Bacillus thuringiensis(Bt)produces multiple insecticidal proteins and serves as a vital biological control agent.This study aims to identify Bt strains with insecticidal activity against lepidopteran pests,thereby alleviating pest resistance pressure and expanding resource reserves.A total of 109 Bt strains were selected from soil of Hebei Province.The shape of parasporal crystals was observed under an oil microscope,and the insecticidal protein genotypes of the Bt strains were identified by PCR using 40 pairs of primers(including cry,cyt,and vip genotypes).The insecticidal activity bioassay was performed using five lepidopteran pests,including Plutella xylostella and Spodoptera litura.The molecular weight of crystal proteins was detected via SDS-PAGE.The shape of parasporal crystals from 90 strains was spherical,and the crystals from the remaining 19 strains were diamond-shaped.The detection rate of insecticidal protein genotypes reached 93.6%,with 29 distinct cry genotypes,1 cyt genotype,and 3 vip genotypes identified.A total of 91 strains possessed combinations of at least two insecticidal protein genotypes,among which 26 strains contained six or more,predominantly exhibiting cry+vip patterns.We found that 19 Bt strains showed high insecticidal activity against various lepidopteran pests such as P.xylostella. These strains primarily expressed proteins at 130,65 ku,encompassing lepidopteran-killing genotypes such as cry1,cry2,and cry15,consistent with PCR identification results.Here,we identified broad-spectrum insecticidal and highly toxic Bt strains against lepidopteran pests,and revealed its insecticidal genes.Our work provides an important candidate for the development of bioinsecticide.

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

In recent years,soil acidification and aluminum ion toxicity in southern cultivated land have intensified.To explore the gene loci related to aluminum tolerance in rapeseed,the F₂ population obtained by crossing the aluminum tolerant variety R248 with the aluminum sensitive variety S120 was used as the experimental material.The extreme mixed pool of aluminum toxicity tolerance was constructed by phenotypic identification,and QTL localization analysis was performed by BSA-seq.The phenotype identification results showed that the aluminum toxicity tolerance of the F₂ population exhibited a continuous normal distribution.After sequencing,mutation detection,and association analysis,a total of 4 intervals related to aluminum toxicity tolerance were obtained,distributed on chromosomes A07,C02,C03,and C08,with a total length of 0.73 Mb and 98 genes.Based on the enrichment analysis results and gene annotation information,it is predicted that BnaA07g35600D,BnaA07g35660D,BnaA07g35700D,BnaA07g3570D, and BnaC03g39670D will participate in the synthesis and transportation of aluminum tolerance hormones in rapeseed,BnaC03g39840D participates in the synthesis of cell wall pectinesterase,BnaC03g39750D and BnaC08g20580D were transcription factors with MYB structure,BnaC03g39850D and BnaC03g39980D were involved in amino acid metabolism.These genes are highly correlated with aluminum tolerance in rapeseed,laying the foundation for the cloning and functional research of aluminum tolerance genes in the next step.

This study seeks to develop a highly efficient dual-base editing tool for solanaceous crops to address the challenges of poor compatibility and low editing efficiency associated with existing base editing systems in these plants. The research involved fusing the nickase Cas9 enzyme (nCas9),which has partially lost its nuclease function,with components such as the highly efficient adenine deaminase ABE8e,the optimized cytosine deaminase sdd7-mini,and the uracil glycosylation inhibitor UGI. Systematic optimization was conducted based on the promoter preferences and codon usage frequencies specific to solanaceous crops. Ultimately,the dual-base editor TPDBE-S was constructed. In transient transformation experiments using tomato and eggplant protoplasts,this editor exhibited exceptionally high editing efficiency. In the stable transformation system of tobacco,targeting the PDS gene verification revealed that four out of five positive transgenic families exhibited pronounced albinism phenotypes,thereby confirming the stable editing capability. The editing window of this editor aligns closely with the characteristics of ABE8e and sdd7-mini. Furthermore,optimizing codon preference and promoter activity in Solanaceae crops significantly enhances editing efficiency and adaptability. The construction of TPDBE-S employs a modular design. The multi-target expression cassette,based on the tRNA self-cleaving mechanism,can be rapidly assembled through one-step gene synthesis,which is both cost-effective and straightforward. This tool addresses the gap in double-base editing specifically for Solanaceous crops.

To clarify the genetic characteristics of total leaf number and leaf number above the ear in maize,and reveal their regulatory effects on maize yield potential,biomass accumulation,and lodging resistance,experiments on QTL mapping and gene mining for controlling maize leaf number were conducted.By using SLAF-seq technology for high-throughput sequencing of Chang 7-2,PHB1M,and 138 F_2∶3 families,combined with two planting density treatments(E1:60 000 plants/ha;E2:120 000 plants/ha)QTL mapping was performed on leaf number phenotypic data,and gene mining was performed on the mapping results.The results showed that two total leaf number QTLs distributed on chromosome 8 were the main effect QTLs,contribution rates were 16.96% and 23.08%,respectively,and the additive effect value was negative;the QTL for the leaf number above the ear distributed on chromosomes 2 and 4,with a positive additive effect value.the two QTLs distributed on chromosome 4 were the main effect QTL,contribution rates were 10.18% and 14.75%,respectively;the QTLs for total leaf number and leaf number above the ear were distributed in different chromosomal regions and might be subject to relatively independent genetic regulation.Gene functional analysis revealed genes screened involvement in carbohydrate metabolism,plant hormone signal transduction,and selected some transcription factors.The results of this study provide richer theoretical support for further revealing the genetic basis of maize leaf number and offer targets for marker-assisted breeding.

To investigate the structure and function of the potato molybdenum cofactor sulfurase(LOS5)gene,StLOS5 was cloned from the potato cultivar Zicai No.3 using PCR,followed by bioinformatics analysis,subcellular localization assays,yeast stress-tolerance assays,and expression pattern analysis under drought stress.The results showed that the CDS region of the potato StLOS5 gene was 2 460 bp in length,encoding 819 amino acids,with a molecular weight of 91.50 ku,a theoretical isoelectric point of 6.78,and a GRAVY value of -0.268,indicating that the protein was hydrophilic.Phylogenetic analysis revealed that the StLOS5 protein had the highest homology with tomato LOS5,suggesting that the two proteins may possess similar functions.Multiple cis-acting elements related to abiotic stress were detected in the promoter region.Subcellular localization showed that StLOS5 was primarily localized in the nucleus.Yeast stress-tolerance assays indicated that StLOS5 enhances tolerance to osmotic stress.Quantitative Real-time PCR analysis demonstrated that StLOS5 exhibits tissue-specific expression,with higher transcript levels in leaves,and responds rapidly to drought stress,with its expression in leaves peaking 12 h after treatment with 8%PEG.These results indicated that StLOS5 plays a role in the potato response to drought stress.

Aquaporins(AQPs)are primarily responsible for the transmembrane transport of water molecules and are involved in processes such as plant growth,development, and responses to environmental stress. Garlic is an important vegetable crop widely cultivated worldwide, but there are currently few reports on salt-tolerance genes in garlic.It aimed to clone the genes AsPIP1;1 and AsTIP2;1, encoding plasma membrane intrinsic protein (PIP) and tonoplast intrinsic protein (TIP) respectively, from garlic, analyze their sequence characteristics, investigate their expression patterns under salt stress conditions, and evaluate their potential roles in garlic salt tolerance. We cloned the AsPIP1;1 and AsTIP2;1 genes from garlic using RT-PCR. Bioinformatics methods were employed to analyze the open reading frames, encode amino acid sequences, and conserve domains of the cloned genes. Furthermore, the expression profiles of AsPIP1;1 and AsTIP2;1 in different tissues and under salt stress conditions were detected using Quantitative Real-time PCR. The results showed that the open reading frames of the AsPIP1;1 and AsTIP2;1 genes were 867, 747 bp in length, encoding 288,248 amino acids, respectively, with the conserved NPA motifs. AsPIP1;1 and AsTIP2;1 were highly expressed in garlic leaves and roots, and salt stress strongly induced changes in the transcriptional levels of both genes. Furthermore,AsPIP1;1 and AsTIP2;1 may interact with other AQP proteins, cell wall-associated proteins, and transporter proteins to adapt to the salt stress environment. These results indicate that the AsPIP1;1 and AsTIP2;1 genes may be involved in the garlic plant's response to salt stress.

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

To investigate the genetic evolution and molecular differences of Lentinula edodes germplasm resources and their hybrid progeny,it utilized whole-genome resequencing technology and analyzed genetic diversity and population structure based on single-nucleotide polymorphism(SNP)data.It involved eight main cultivated varieties,six hybrid new varieties,and two wild strains of L.edodes.The results revealed that the SNP numbers in the wild strains Y5 and Y6 were significantly higher than those in the cultivated varieties,with 158 659 and 149 422 SNPs,respectively,and they clustered in independent branches based on genetic distance.In contrast,the cultivated varieties and their hybrid progeny exhibited similar SNP numbers and genetic relationships,with SNP counts ranging from 97 295 to 105 627 and genetic distances ranging from 0.001 2 to 0.055 3,clustering within the same branch.The branches of the cultivated varieties and their hybrid progeny could be divided into six closely related groups.These included early-maturing varieties L18 and 868,early-maturing white-faced varieties RX11,QK212,and 0912,medium-maturing firm-textured varieties 808,LX1,and 168,hybrid progeny varieties JX15 and JX3 of 808,L18,and 868,hybrid progeny varieties JXB5 and JXB15 of JX15 and JX3,and hybrid progeny varieties E14 and E36 of 0912 and JXB5.Population structure and principal component analysis revealed that L.edodes germplasm resources could be divided into four subgroups.The first class included varieties RX11,QK212,and 0912,as well as their hybrid progeny E14 and E36.The second class consisted of main cultivated varieties LX1,168,and 808,along with their hybrid derivatives JX15 and JX3,which share extremely similar genetic information or background.The third class comprised L18 and 868,along with their multiple-generation,genetically improved,high-quality hybrid strains,JXB5 and JXB15,which exist in a transitional state between cultivated varieties and wild strains,with a propensity towards the cultivated varieties.The fourth class was represented by wild strains YX5 and YX6,which are distinctly different from the cultivated strains.This study clarified the genetic differences among L.edodes strains,laying the foundation for the selection of parents and the pairing of hybrid combinations in subsequent hybrid breeding.

In order to elucidate the molecular mechanism of flour-quality regulation at the transcriptional level,wheat mutant varieties A94 and A261 were obtained by ⁶⁰Co-γ irradiation mutagenesis of Shimai 15,and the developing seeds at days 7,14,21 and 28 after flowering were subjected to transcriptome sequencing,and water absorption,dough development time,stable time,softening degree and farinogram index of the three varieties were measured.The results showed that farinogram parameters of two mutant varieties were better than those of Shimai 15.Twenty-four samples were sequenced,and after the data were filtered and annotated,a total of 26 714 genes were identified.With respect to Shimai 15,there were 1 042,258,301 and 366 differentially expressed genes(DEGs)at four time points in A94,respectively,and there were 2 178,248,117 and 959 DEGs in A261,respectively.KEGG enrichment showed that DEGs were mainly involved in pathways of starch and sucrose metabolism,protein processing in endoplasmic reticulum.GO enrichment showed that DEGs were mainly involved in glycogen biosynthesis processes,amyloplast and other biological processes.By analyzing the differential genes in these pathways and biological processes,it was hypothesized that the reasons for the improved flour quality in the mutant varieties include the up-regulation of genes related to sucrose and starch synthesis and metabolism in the early stages of filling,the degradation of misfolded proteins in the middle and late stages,and the differential expression of genes regulating protein and starch accumulation.

Wheat leaf rust and wheat powdery mildew are two important diseases in the world.Cultivating and planting disease resistant varieties is the most economical and effective method to control these two diseases.In order to test the distribution of wheat leaf rust resistance gene Lr21,the molecular marker closely linked to Lr21 was used to detect 1 200 wheat varieties(lines),and 23 wheat varieties(lines)were detected to contain Lr21.The molecular markers related to other leaf rust resistance genes were used to detect 23 wheat materials,Lr20 gene was detected in Gaoyou 2018,Lr37 gene was detected in Aifeng 8,Jingdong 22,Zhongmai 175,Luyuan 205 and Tang Y958,and Lr46 gene was detected in Jinan 17.No wheat varietie(line)was detected to contain Lr9,Lr10,Lr19,Lr24 or Lr34.Resistance evaluation of 23 wheat varieties(lines)containing Lr21 was identified using the epidemic races of Puccinia triticina(Pt)and Blumeria graminis f.sp.tritici(Bgt),respectively.The results showed that seven wheat varieties(lines)were resistant to both Pt and Bgt,with a frequency of 30.43%,and six wheat varieties(lines)were susceptible to Pt and Bgt mildew at the same time,and the frequency was 26.09%.Taken together,Lr21 was detected in 1 200 wheat cultivars(lines),as well as the presence of other rust resistance genes,and further evaluated their resistance to Pt and Bgt,which will provide a theoretical basis for the selection of multi-resistant wheat.

Bacillus subtilis ZA1 exhibits notable antibacterial and growth-promoting activities. Helicases have been established as crucial protein complexes in nucleic acid metabolism, with members of the helicase superfamily performing key catalytic functions during genetic information transfer by specifically cleaving phosphodiester bonds in DNA/RNA molecules. To elucidate the potential roles of the helicase genes ypvA and yjcD in the antibacterial process of B.subtilis ZA1, we used B.subtilis ZA1 as the experimental material, cloned the ypvA and yjcD genes, analyzed their sequence characteristics through bioinformatics, and determined their subcellular localization. A preliminary investigation was conducted on the structural and functional properties of these two genes and their encoded proteins. The results showed that the cloned coding sequences (CDS) of ypvA and yjcD were 1 791, 1 767 bp in length, encoding 596, 588 amino acids, respectively. The predicted relative molecular masses of the encoded proteins were 68.804 01, 68.275 08 ku, with theoretical isoelectric points (pI) of 4.81, 8.25, respectively, and both exhibited weak hydrophilicity. The spatial structures of the ypvA and yjcD proteins consisted of a compact architecture primarily composed of α-helices, β-sheets, and extensive random coils. Subcellular localization analysis indicated that both proteins were localized in the cytoplasm and nucleus, lacking transmembrane domains and signal peptides, thus classifying them as non-secretory proteins. Analysis of conserved domains revealed that ypvA belonged to the DinG family of Rad3-related DNA helicases, while yjcD belongs to the UvrD superfamily I DNA/RNA helicases. Further phylogenetic analysis confirmed that both proteins are ATP-dependent helicases. In summary, cloning and sequence analysis of the ypvA and yjcD genes from B.subtilis ZA1 identified them as non-secretory ATP-dependent helicases.

The study aimed to explore the genetic mechanism of fruit shape in oriental melon.The parents,M125(Elongate)and M30(Near round),were used to build the six generation populations,and the genetic patterns of fruit longitudinal diameter,transverse diameter,and shape index were studied using the method of main gene+multiple gene generation analysis.The results showed that significant positive correlation between fruit longitudinal diameter and shape index,and the correlation values were high in 2021-2023.In three years,the genetic models of fruit longitudinal diameter,transverse diameter,and shape index were MX2-ADI-ADI,two pairs of additive-dominant-epistatic master genes+additive-dominant-epistatic polygenes.In the first-order genetic parameters,the additive effect values of the two main genes controlling the fruit longitudinal diameter,transverse diameter,and shape index were equal and positive in the three years(d_a=d_b>0),and it had a synergistic effect.The absolute value of the dominant effect of the first pair of major genes controlling fruit longitudinal diameter in 2021 and 2023 was significantly higher than that of the second pair of major genes,while the dominant effect of the second major gene was slightly higher than that of the first pair of major gene in 2022.The additive effects of the two pairs of major genes controlling fruit transverse diameter showed little variation across different years and were all negative,indicating that the dominant effects of the major genes had a negative impact on fruit diameter.The absolute value of the dominant effect of the first pair of major genes controlling fruit shape index was greater than that of the second pair of major genes,indicating that the first pair of major genes played a leading role in the dominant effects.The second-order genetic parameters showed that in the F₂,from 2021 to 2023,the heritability of main genes for fruit longitudinal diameter was 81.51%,72.16%,80.77%,respectively.The heritability of major genes for transverse diameter was 77.88%,69.94%,65.90%,respectively.The heritability of main genes for fruit shape index was 80.41%,70.81%,82.49%,respectively.The inheritance of the three fruit shape traits in Oriental melon is quantitative traits,and the heritability of major genes in F₂ generation was high.Therefore,the three fruit shape traits were selected in early generations during the oriental melon breeding.

Wheat,as one of the world's major food crops,its growth and development,yield,and quality are often constrained by abiotic stresses. To cope with these abiotic stresses,wheat has evolved a series of response mechanisms,including signal perception,signal transduction,and gene expression regulation,to maintain cellular homeostasis and protect the functions of biological macromolecules. Drought stress causes the accumulation of osmotic-adjustment substances and antioxidants in wheat,a decline in the photosynthetic rate,and alterations in morphological structure. Its molecular mechanisms mainly involve pathways such as reactive oxygen species (ROS) signaling,hormone signaling,and gene expression regulation. Saline-alkali stress inhibits seed germination and seedling growth,leading to photosynthetic impairments,osmotic-regulation imbalances,ion-balance disorders,and membrane-lipid oxidation. Under such circumstances,wheat can enhance salt tolerance by activating the salt overly sensitive (SOS) signaling pathway,calcium-ion signaling,hormone signaling,and gene-expression regulation. Extreme temperature stress inhibits photosynthesis,triggers membrane-lipid oxidation,and promotes the accumulation of osmotic-adjustment substances. Its regulatory mechanisms mainly involve hormone signaling,transcriptional activation,and epigenetic regulation. This study reviews the response mechanisms of wheat to drought,saline-alkali,and extreme temperature stresses and looks ahead to future research directions:focusing on the cross-interaction mechanisms of multiple stresses and mining genes responsive to combined stresses; effectively integrating stress-resistant traits into high-yield and high-quality genetic backgrounds to achieve the coordinated improvement of ideal plant types and stress resistance; promoting the in-depth integration of modern biotechnology with conventional breeding methods to enhance breeding efficiency. It aims to provide comprehensive theoretical support and technical references for the genetic improvement of wheat's resistance to abiotic stresses.

bHLH(Basic helix-loop-helix)gene family is one of the largest plant transcription factor families,binds to cis-acting elements on target gene promoters and plays crucial roles in various physiological processes including growth development,secondary metabolism regulation,signal transduction,and stress response.As one of the world's most important cash crops,cotton is severely affected by stresses such as Verticillium and Fusarium wilt,low temperature,high temperature,salinity,drought,and heavy metal exposure,which significantly impact yield and fiber quality.Meanwhile,with the continuous decline in cotton cultivation benefits,the high-value utilization of by-products such as cottonseeds and cotton secondary metabolites plays a crucial role in improving the comprehensive utilization value of cotton.In this paper,the functions of cotton bHLH transcription factors in cotton fiber development,biotic stress response,abiotic stress response,plant architecture,anther,glandular development,and somatic embryogenesis were summarized.Furthermore,it discusses the application prospects of in-depth analysis of the biological functions of bHLH transcription factors in enhancing the utilization value of cotton by-products,such as cottonseed oil,cottonseed protein,gossypol,anthocyanins,cottonseed vitamins,and nectar,in order to provide theoretical reference for further clarifying the functions of bHLH transcription factors in cotton growth and development,as well as improving the comprehensive utilization value of cotton by-products.

Waterlogging stress severely impacts soybean production.To investigate the effects of waterlogging stress on soybean growth and development and elucidate its response mechanisms,this study employed four soybean germplasms cultivated in the Huaibei region as experimental materials.Through simulated waterlogging treatment at the V2 stages,combined with yield analysis,physiological indices,and transcriptomic profiling,the study explores the waterlogging tolerance mechanisms in soybean.The results demonstrated that waterlogging stress significantly reduced dry matter accumulation and nitrogen uptake efficiency in the waterlogging-sensitive cultivar Xudou 18,resulting in a yield loss of 30.23%.In contrast,the waterlogging-tolerant cultivar Huaidou 13 exhibited stronger adaptability with only a 16.77% yield loss,attributed to stabilized root nitrogen absorption and maintained root dry matter accumulation.Transcriptomic analysis revealed that differentially expressed genes(DEGs)in Huaidou 13 under waterlogging stress were significantly enriched in pathways that related to the biosynthesis of secondary metabolites,photosynthetic systems,and antioxidant activity.Conversely,DEGs in Xudou 18 were predominantly associated with hormone transduction,photosynthesis,and peroxisome-related pathways.A total of 148 genotype-specific DEGs were identified between cultivars with contrasting waterlogging tolerance,primarily enriched in photosynthesis,secondary metabolite biosynthesis,and fatty acid metabolism.Identified transcription factors included members of the AP2/ERF-ERF,C3H,ARR-B,NAC,and WRKY families.In summary,transcription factors or genes in secondary metabolic pathways that may be related to waterlogging tolerance were screened.

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.