甜樱桃扩展蛋白PavEXPA2基因克隆与表达分析
作者:
作者单位:

贵州大学生命科学学院/农业生物工程研究院/ 山地植物资源保护与保护种质创新教育部重点实验室,贵阳 550025

作者简介:

张焱,E-mail:1694291562@qq.com

通讯作者:

文晓鹏,E-mail:xpwensc@hotmail.com

中图分类号:

S662.5

基金项目:

国家自然科学基金项目(32272649);贵州省教育厅重点项目(黔科合KY字[2021]038)


Cloning and expression of expansin PavEXPA2 gene in sweet cherries
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Affiliation:

The Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education)/Institute of Agro-Bioengineering/College of Life Sciences, Guizhou University, Guiyang 550025, China

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    摘要:

    为揭示甜樱桃中扩展蛋白基因的结构与功能,克隆了甜樱桃PavEXPA2基因,并分析了该基因在不同组织(茎、花芽、盛开花朵、幼叶、老叶、幼叶叶柄、老叶叶柄、2个脱落高峰期正常果柄与即将脱落果柄、2个脱落高峰期正常果实与脱落果实)及逆境胁迫(使用20% PEG6000和20 mmol/L NaCl溶液分别模拟干旱和盐胁迫,分别处理0、2、4、6、8 h)下的表达情况。结果显示:PavEXPA2基因cDNA全长序列为1 035 bp,开放阅读框(ORF)为852 bp,编码283 aa,蛋白等电点为8.90,分子质量约为30.81 ku,含有2个跨膜螺旋结构和1个信号肽,亚细胞定位预测PavEXPA2定位于细胞壁;组织表达分析显示,PavEXPA2基因在甜樱桃易脱落组织中表达量较高,如盛开花瓣,即将脱落的果柄、果实、叶柄等,其中在即将脱落叶柄中表达量最高;相对于未处理样品,干旱处理样本PavEXPA2表达量上升,在处理6 h时表达量达到最高,随后降低;盐胁迫条件下,PavEXPA2表达量呈先下降后上升再下降的趋势,其中在处理6 h时表达量达到最高。结果表明,甜樱桃可能通过PavEXPA2基因上调促进脱落而抵御逆境胁迫。

    Abstract:

    Expansin is a kind of protein widely found in plants.It may relax the cell wall and plays an important role in the growth and development,and the stress response of plants.The PavEXPA2 gene was cloned and bioinformatically analyzed to uncover the structure and function of expansingene(EXPA) in sweet cherries.The gene expression in different tissues including stems,flower buds,blooming flowers,young leaves,old leaves,petioles of young leaves,petioles of old leaves,normal fruit stalks at the 2 abscission peaks and about to abscission,the normal fruits at 2 abscission peaks and shedding,and under the abiotic stresses including using 20% PEG6000 and 20 mmol/L NaCl solutions to simulate drought and salt stress for 0,2,4,6,and 8 h was profiled.The results showed that the full length of PavEXPA2 was 1 035 bp with the open reading frame (ORF) of 852 bp,encoding 283 amino acids.The putative protein has an isoelectric point of 8.90 and a molecular mass of 30.81 ku,containing two transmembrane helical structures and a signal peptide.The results of subcellular location showed that the protein of gene was located in the cell wall.The results of tissue expression analysis showed that the expression level of PavEXPA2 was higher in the easily abscised tissues of sweet cherry including flowers,stalks,fruits and petioles,with the highest level in petioles about to abscission.Compared with the samples untreated,the expression of PavEXPA2 in the drought-treated samples increased,and the expression reached the highest at 6 h,and then decreased.Under the salt stress,the expression level of PavEXPA2 decreased first,then increased and then decreased,and the expression level reached the highest at 6 h.Therefore,sweet cherries may resist stress by promoting abscission through the up-regulation of PavEXPA2 gene.It will lay a theoretical foundation for further understanding the role of PavEXPA2 in the growth and development of sweet cherries.

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张焱,仇志浪,申洛男,文晓鹏.甜樱桃扩展蛋白PavEXPA2基因克隆与表达分析[J].华中农业大学学报,2023,42(1):27-33

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  • 收稿日期:2022-02-22
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  • 在线发布日期: 2023-02-22
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