山新杨谷胱甘肽S-转移酶编码基因PdbGSTU功能分析

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国家科技期刊平台
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中文摘要：[目的]本研究旨在通过分析山新杨谷胱甘肽S-转移酶编码基因PdbGSTU的抗病功能，为林木抗性育种提供基因资源与抗性种质。[方法]克隆PdbGSTU基因序列并对其进行生物信息学分析，利用荧光定量PCR技术分析该基因的组织特异性表达及植物激素诱导下的表达模式。利用转基因技术获得山新杨的过/抑制表达PdbGSTU基因植株，通过观察比较接种细链格孢菌后各植株叶片的表型和病斑面积，验证该基因的抗病功能;同时测定接种病原菌前后，野生型和转基因山新杨植株内过氧化氢含量和抗氧化相关酶活性。[结果](1)山新杨PdbGSTU基因开放阅读框全长753 bp，编码氨基酸250个，对应的蛋白质相对分子质量为29。01 kDa，为稳定的酸性亲水蛋白，定位于细胞质中;系统进化分析显示，PdbGSTU蛋白与银中杨的蛋白KAJ6918316亲缘关系最近;启动子序列分析显示，PdbGSTU基因启动子序列含多种响应植物激素和逆境胁迫的顺式作用元件。(2)RT-qPCR结果显示，PdbGSTU基因在山新杨顶芽表达量最高，在其根部表达量最低，且该基因受茉莉酸甲酯、水杨酸和1-氨基环丙基-1-羧酸3种植物激素诱导，均上调表达。(3)接种细链格孢菌后，野生型和抑制表达PdbGSTU基因植株的叶片上，病斑面积分别为6。42和16。46 mm2，而过表达PdbGSTU基因的植株叶片上，少部分接种点出现明显病斑，其余接种部分仅出现褪色。[结论]PdbGSTU正向参与山新杨对细链格孢菌侵染的抵御过程，可通过清除活性氧提高杨树对病原菌的抗性。

外文标题：Functional analysis of encoding gene PdbGSTU for glutathione S-transferase in Populus davidiana × P.bolleana

外文摘要：[Objective]This paper aims to provide genetic resources and resistant germplasm for tree resistance breeding by analyzing the disease-resistance function of encoding gene PdbGSTU for glutathione S-transferase in Populus davidiana × P. bolleana.[Method]PdbGSTU was cloned in P. davidiana × P. bolleana by PCR,and bioinformatics analysis were analyzed. The tissue specific expression patterns and expression patterns induced by plant hormones of PdbGSTU were also analyzed by RT-qPCR. Overexpressing/sciencing PdbGSTU plants of P. davidiana × P. bolleana obtained through transgenic technology and function of PdbGSTU on disease resistance were verified by observing and comparing the phenotypes and concurrent lesion areas of leaves after inoculation with Alternaria alternata. Moreover,the contents of H2O2 and the activities of antioxidant enzyme were measured in leaves before and after A. alternata inoculation.[Result](1) The open reading frame (ORF) length of PdbGSTU was 753 bp,which encoded 250 amino acids with the protein relative molecular mass 29.01 kDa. The PdbGSTU was a stable hydrophilic acidic protein located in the cytoplasm. Systematic evolutionary analysis revealed that PdbGSTU in P. davidiana × P. bolleana had a close relationship with KAJ6918316 in P. alba × P. berolinensis. The analysis of cis-acting elements in promoter sequence of PdbGSTU showed that,this sequence contained multiple cis-acting elements related to plant response to stress or phytohormones. (2) RT-qPCR showed that PdbGSTU had the highest expression level in apical bud of P. davidiana × P. bolleana,and the lowest expression level in root. In addition,PdbGSTU could response to MeJA,SA and ACC,and show up-regulated expression. (3) After inoculation with A. alternata,the lesion areas on the leaves of wild-type and PdbGSTU gene suppressed P. davidiana × P. bolleana were 6.42 and 16.46 mm2,respectively. However,on the leaves of plants overexpressing PdbGSTU gene,a small number of inoculation sites showed obvious lesions,while the rest of inoculation sites only showed fading.[Conclusion]PdbGSTU is positively involved in the resistance process of P. davidiana × P. bolleana to A. alternata infection,and can enhance the resistance of P. davidiana × P. bolleana to pathogens by clearing reactive oxygen species.

外文关键词：

gene expressionPopulus davidiana × P. bolleanaglutathione S-transferasesAlternaria alternatadisease resistance function

作者：

黄颖、王晓东、遇文婧

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作者单位：

黑龙江省森林保护研究所,黑龙江哈尔滨 150080

东北林业大学林学院,林木遗传育种全国重点实验室,黑龙江哈尔滨 150040

关键词：

基因表达山新杨谷胱甘肽S-转移酶细链格孢菌抗病功能

基金：

黑龙江省省属科研院所科研业务费项目黑龙江省省属科研院所科研业务费项目中央高校基本科研业务费专项资金项目黑龙江省博士后基金项目

项目编号：

CZKYF2022-1-C048LKSB2024-22572020BA08

出版年：

2024

DOI：

10.12171/j.1000-1522.20240020

北京林业大学学报

北京林业大学

北京林业大学学报

CSTPCD北大核心

影响因子：1.237

ISSN：1000-1522

年,卷(期)：2024.46(8)