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Abiotic stress treatment reveals expansin like A gene OfEXLA1 improving salt and drought tolerance of Osmanthus fragrans by responding to abscisic acid

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Horticultural Plant Journal 2024年第2期10卷 573-585页

作者： Bin Dong Qianqian Wang Dan Zhou Yiguang Wang Yunfeng Miao Shiwei Zhong Qiu Fang Liyuan Yang Zhen Xiao Hongbo Zhao School of Landscape Architecture Zhejiang Agriculture and Forestry UniversityHangzhouZhejiang 311300China Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants HangzhouZhejiang 311300China Key Laboratory of National Forestry and Grassland Administration on Germplasm Innovation and Utilization for Southern Garden Plants HangzhouZhejiang 311300China

Sweet osmanthus(Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental stresses. Currently, limited information is available regarding the genetic analysis and functional identification of expansin genes in response to abiotic stress in sweet osmanthus. In this study, a total of 29 expansin genes were identified and divided into four groups by genome-wide analysis from the sweet osmanthus genome. Transcriptome and quantitative Real-time PCR analysis showed that the cell wall-localized protein expansin-like A(OfEXLA1) gene was significantly induced by salt and drought treatment. Histochemical GUS staining of transgenic Arabidopsis lines in which GUS activity was driven with the OfEXLA1 promoter, GUS activity was significantly induced by salt, drought, and exogenous abscisic acid(ABA). In yeast, we found OfEXLA1overexpression significantly improved the population of cells compared with wild-type strains after NaCl and polyethylene glycol(PEG)treatment. Additionally, OfEXLA1 overexpression not only promoted plant growth, but also improved the salt and drought tolerance in Arabidopsis. To gain insight into the role of ABA signaling in the regulation of OfEXLA1 improving abiotic tolerance in sweet osmanthus, four differentially expressed ABA Insensitive 5(ABI5)-like genes(OfABL4, OfABL5, OfABL7, and OfABL8) were identified from transcriptome, and dualluciferase(dual-LUC) and yeast one hybrid(Y1H) assay showed that OfABL4 and OfABL5 might bind to OfEXLA1 promoter to accumulate the OfEXLA1 expression by responding to ABA signaling to improve abiotic tolerance in sweet osmanthus. These results provide the information for understanding the molecular functions of expansin-like A gene and molecular breeding of sweet osmanthus in future.

关键词： Osmanthus fragrans Abiotic tolerance expansin Abscisic acid

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Overexpression of the apple expansin-like gene MdEXLB1 accelerates the softening of fruit texture in tomato

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Journal of Integrative Agriculture 2022年第12期21卷 3578-3588页

作者： CHEN Yan-hui XIE Bin AN Xiu-hong MA Ren-peng ZHAO De-ying CHENG Cun-gang LI En-mao ZHOU Jiang-tao KANG Guo-dong ZHANG Yan-zhen Key Laboratory of Mineral Nutrition and Efficient Fertilization for Deciduous Fruits Liaoning Province/Key Laboratory of Fruit Germplasm Resources UtilizationMinistry of Agriculture and Rural Afffairs/Institute of PomologyChinese Academy of Agricultrual SciencesXingcheng 125100P.R.China National Engineering Research Center for Agriculture in Northern Mountainous Areas/Mountainous Area Research Institute Hebei Agricultural UniversityBaoding 071001P.R.China

Fruit firmness is an important quality trait of apple fruit texture,and the pre-harvest ripening period is the key period for the formation of apple fruit texture.expansin is a cell wall loosing protein family that has four subfamilies:α-expansin(EXPA),β-expansin(EXPB),expansin-like A(EXLA),and expansin-like B(EXLB).In this study,we investigated the key period of pre-harvest texture formation in‘Golden Delicious’apples based on fruit longitudinal diameter,transverse diameter,firmness,tissue structure,respiration intensity,ethylene release rate,and expansin activity.Within the 10 days before harvest,the fruit was found to reach maturity.Semi-quantitative RT-PCR revealed that most of the expansins were expressed at the ripening stage before harvest.The biological function of the EXLB subfamily gene,MdEXLB1,was further identified,and its subcellular localization on the cell wall was confirmed by transient transformation experiments.Compared with the wild type(WT),the transgenic tomato lines overexpressing MdEXLB1 had lower plant height,earlier fruiting period,fewer days for fruit ripening,higher fruit maturity,lower fruit firmness,higher fruit expansin activity,more discrete flesh cell structure,and accelerated fruit ripening process.Overall,this is the first study to propose that the apple EXLB subfamily gene,MdEXLB1,has biological functions and plays an important role in promoting fruit ripening and softening.

关键词： fruit firmness expansin apple transgenic tomato

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expansin Genes Expressed Differentially in Peduncle Elongation of Near-Isogenic Wheat Rht Lines

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Journal of Integrative Agriculture 2014年第10期13卷 2091-2101页

作者： WANG Jia-li LIU Dong-cheng GUO Xiao-li YANG Wen-long ZHANG Ai-min State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental BiologyChinese Academy of Sciences Institute of Shandong River Wetlands College of Biology China Agricultural University The Collaborative Innovation Center for Grain Crops in Henan College of AgronomyHenan Agricultural University

The introduction of reduced height （Rht） genes Rht-Blb and Rht-Dlb led to impressive increases in wheat （Triticum aestivum L.） yields during the Green Revolution. In the present study, the dynamic elongation of peduncle in a set ofnear-isogenic lines （NILs） carrying different Rht alleles （Rht-Blb, Rht-Dlb, Rht-Blc, Rht-Dlb＋Rht-Blb, and Rht-Dlb＋Rht-Blc） were investigated. The reduction of the final length of peduncle in NILs was dependent mainly on the elongation rate, which was reduced by Rht genes, during rapid elongation phase. Resin sections showed that Rht genes strongly reduced the cell extension in peduncle. The expression of expansin genes, which mediate cell wall loosening and leading to cell expansion, were analysed by using real- time quantitative PCR （qPCR）. Among the 23 possible wheat expansin genes, 17 were expressed in the peduncle. The spatial distribution of expression was further analysed for five expansins that showed high expression levels in the peduncles of Rht lines. Compared to wild type plants, the incorporation of Rht-Dlb allele decreased about 37 and 80% of the expression levels of ExpA 7 and ExpA3 in elongation zone, respectively. The presence Rht-Blc dwarfing genes, however, produced 53% reduction in the expression level of ExpA7, and seriously decreased about 70% of ExpB9 expression. Although the expression levels of five genes exhibited variability among the lines, an expansin gene, ExpB2, showed its expression level highly associated with the cell elongation rate in peduncle of different Rht lines.

关键词： expansin peduncle gene expression dwarfing gene

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Cloning and expression analysis of a homologous expansin gene EXP2 in Picea wilsonii

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Journal of Forestry Research 2016年第2期27卷 247-255页

作者： Tong Zhang Yanfang Li Yanni Zhou Lingyun Zhang Key Laboratory of Forest Silviculture and Conservation of the Ministry of Education Beijing Forestry University

expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends based on the cDNA library of Picea wilsonii and EST fragment of PwEXP2. It was found that PwEXP2 coded 253 amino acids, and putative signal peptides exist at the N-terminal, followed by 8 cysteines, a HFD （His-Phe-Asp） conserved domain, and 4 tryptophan residues at the C-terminal. PwEXP2 was located in cytoplasm and nucleus when transformed in an onion epidermal cell. Quantitative real-time PCR assays showed that PwEXP2 was expressed in various tissues with a relatively high level in needles and low level in mature pollen. The expression level of PwEXP2 dramatically increased after seed germination. Gene expression profiles in abiotic stresses showed that PwEXP2 was induced by high temperature and osmotic stress but not involved in ABA-dependent signaling path- way. These results display the important roles of the PwEXP2 in plant development and multiple adversity stresses.

关键词： expansin Picea wilsonii Stress response Gene expression

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枯草芽孢杆菌expansin蛋白的原核表达及活性分析

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中国生物制品学杂志 2019年第9期32卷 977-982页

作者：叶凯霞朱华琛赵庆新陈洪生南京工业大学生物与制药工程学院江苏南京211800 盐城师范学院盐城市蛋白工程技术研究中心江苏盐城224051 伦敦学院大学医学部伦敦WC1E6BT 江苏博益生物科技有限公司江苏盐城224051

目的原核表达枯草芽孢杆菌expansin蛋白,并分析其促进多糖糖化的活性。方法以过夜培养的Bacillus subtilis subsp. Subtilis subtilis str. 168基因组为模板合成expansin基因;在引物的N-端添加6-his tag,PCR合成融合基因6his-expansin,构建重组质粒6his-expansin-6his-p ET28a(+)[heh-pET28a(+)],转化大肠埃希菌BL21(DE3),IPTG诱导表达融合蛋白6His-expansin-6His(HEH);并对表达条件进行优化。产物经Ni-NTA纯化,检测其与多糖降解酶(polysaccharide degrading enzyme,PDE)(纤维素酶、木聚糖酶、果胶酶)及多糖复合酶(polysaccharide complex enzyme,PCE)制剂的协同活性(synergistic activity,SA)。结果经双酶切鉴定,重组质粒heh-p ET28a(+)构建正确;融合蛋白HEH在0. 05 mmol/L IPTG 30℃诱导30 h的最适表达条件下,表达量约为1. 2 mg/m L,其相对分子质量为26 400,目的蛋白纯度达95%;该蛋白与纤维素酶、木聚糖酶、果胶酶的SA分别为171. 53%、61. 95%、230. 00%,与PCE降解滤纸(filter paper)、秸秆(rice straw)的SA分别为312. 83%、356. 59%。结论重组的融合蛋白HEH与PDE均具有较高的协同作用,为expansin在生物质木质纤维素糖化领域的应用奠定了基础。

关键词：枯草芽孢杆菌 expansin 原核细胞基因表达多糖糖化协同活性

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expansin超级家族的进化与命名

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广东农业科学 2007年第8期34卷 133-135页

作者：牛艳梅沈文涛周鹏华南热带农业大学热带生物技术研究所/热带生物技术国家重点实验室海南海口571101

综述了expansin基因家族的命名法和进化研究的最新进展,介绍了expansin超级家族的发现过程和定义,分析了其基因结构和进化状况,详述了expansin这个超级家族成员的命名法则。

关键词： expansin 进化命名

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Cloning and Sequence Analysis of expansin Genes from Litchi chinensis Fruit

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Agricultural Sciences in China 2003年第6期2卷 658-662页

作者： LUWang-jin JIANGYue-ming CollegeofHorticulture SouthChinaAgriculturalUniversityGuangzhou510642P.R.China SouthChinaInstituteofBotany ChineseAcademyofSciencesGuangzhou510650P.R.China

Using PCR degenerate primers, designed with reference to the sequences of the conserved amino acids of known expansins, to amplify cDNA fragments in litchi fruit by RT-PCR, two different cDNA fragments , named as Lc-Exp1 and Lc-Exp2 , were cloned. Lc-Exp1 and Lc-Exp2 was respectively composed of 531 bp encoding 177 amino acids and 537 bp encoding 179 amino acids. Eight cysteine residues and three tryp-tophan residues, which is supposed to be the characteristics of expansins, are conserved in both Lc-Exp1 and c-Exp2. In addition, the homology between the two expansins is 71. 6% at nucleotide acid sequences and 76.3% at amino acid sequences. The homology of Lc-Exp1 with Fa-Exp2 or Pp-Exp1 was 92.7% or 92.1%, but that of Lc-Exp2 with Fa-Exp2 or Pp-Exp1 was only 77. 4% or 76.3% at amino acid sequences.

关键词： Litchi chinensis Sonn. fruit expansin cDNA cloning Sequence analysis

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Exp2 polymorphisms associated with variation for fiber quality properties in cotton(Gossypium spp.)

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The Crop Journal 2014年第5期2卷 315-328页

作者： Daohua He Zhongping Lei Hongyi Xing Baoshan Tang Junxing Zhao Bixia Lu College of Agronomy Northwest A&F University College of Life Sciences Northwest A&F University

Plant expansins are a group of extracellular proteins thought to affect the quality of cotton fibers. Previous expression profile analysis revealed that six expansin A genes are present in cotton, of which two(GhExp1 and GhExp2) produce transcripts that are specific to the developing cotton fiber. To identify the phenotypic function of Exp2, and to determine whether nucleotide variation among alleles of Exp2 affects fiber quality, candidate gene association mapping was conducted. Gene-specific primers were designed to amplify the Exp2 gene. By amplicon sequencing, the nucleotide diversity of Exp2 was investigated across92 accessions(including 7 Gossypium arboreum, 74 Gossypium hirsutum, and 11 Gossypium barbadense accessions) with different fiber qualities. Twenty-six SNPs and seven InDels including 14 from the coding region of Exp2 were detected, forming twelve distinct haplotypes in the cotton collection. Among the 14 SNPs in the coding region, five were missense mutations and nine were synonymous nucleotide changes. The average SNP/InDel per nucleotide ratio was 2.61%(one SNP per 39 bp), with 1.81 and 3.87% occurring in coding and non-coding regions, respectively. Nucleotide and haplotype diversity across the entire Exp2 region was 0.00603(π) and 0.844, respectively, and diversity in non-coding regions was higher than that in coding regions. For linkage disequilibrium(LD), the mean r2 value for all polymorphism loci pairs was 0.48, and LD did not decay over 748 bp. Based on132 simple sequence repeat(SSR) loci evenly covering 26 chromosomes, the population structure was estimated, and the accessions were divided into seven groups that agreed well with their genomic origin and evolutionary history. A general linear model was used to calculate the Exp2-wide diversity–trait associations of 5 fiber quality traits, considering population structure(Q). Four SNPs in Exp2 were associated with at least one of the fiber quality traits, but not with fiber elongation. The highest positive effect on UHML and STR was observe

关键词： Cotton expansin Fiber quality Linkage disequilibrium Association mapping

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植物细胞壁松弛因子——expansin研究进展

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江苏农业科学 2013年第6期41卷 11-13页

作者：张安曹清河周志林赵冬兰李昂李元元唐君江苏徐州甘薯研究中心江苏徐州221121 中国农业科学院研究生院北京100081

就expansin的发现与来源、基因家族分类与基因结构、功能、作用机理等方面展开论述,旨在让人们系统地了解expansin的研究进展,并为未来通过调控expansin的表达来调控植物的生长发育、提高作物产量、延长贮藏时间等生产应用提供参考。

关键词： expansin 扩张蛋白膨胀素细胞壁研究进展

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香蕉果实expansin cDNA克隆及序列分析

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华南农业大学学报 2003年第3期24卷 40-42页

作者：陆旺金华南农业大学广东省果蔬保鲜重点实验室广东广州510642

提取成熟香蕉果肉的RNA并分离mRNA,以mRNA反转录合成cDNA,以该cDNA为模板,设计expansin的简并引物,进行RT PCR(退火温度为50℃).得到的扩增产物纯化后与pGEM T Easy载体连接,转化大肠杆菌,任意挑选2个阳性克隆,进行序列分析,结果得到2... 详细信息

提取成熟香蕉果肉的RNA并分离mRNA,以mRNA反转录合成cDNA,以该cDNA为模板,设计expansin的简并引物,进行RT PCR(退火温度为50℃).得到的扩增产物纯化后与pGEM T Easy载体连接,转化大肠杆菌,任意挑选2个阳性克隆,进行序列分析,结果得到2个序列相同的expansin的cDNA基因片段,命名为MA Exp3(以示与已登录的香蕉MA Exp1和MA Exp2的区别).MA Exp3中存在expansin基因的保守区域,即8个半胱氨酸残基和3个色氨酸残基,它编码177个氨基酸,与MA Exp1的核苷酸同源性为74 2%,氨基酸同源性为86 4%.

关键词：香蕉果实 expansin cDNA克隆序列分析大肠杆菌阳性克隆基因片段核苷酸同源性氨基酸同源性

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