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.

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

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

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

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

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

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

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

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

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

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

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

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

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