研究报告

海岛棉几丁质酶基因GbCHI的克隆与功能分析

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  • 1. 山东农业大学农学院, 作物生物学国家重点实验室, 泰安271018 2.中国科学院微生物研究所, 植物基因组学国家重点实验室, 北京100101

收稿日期: 2011-03-22

  修回日期: 2011-05-23

  网络出版日期: 2012-02-25

基金资助

国家自然科学基金项目(编号:30671116)资助

Cloning and functional analysis of chitinase gene GbCHI from sea-island cotton (Gossypium barbadense)

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  • 1. State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai’an 271018, China 2. State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China

Received date: 2011-03-22

  Revised date: 2011-05-23

  Online published: 2012-02-25

摘要

几丁质酶是植物主要的病程相关(PR)蛋白之一。前期工作中利用比较蛋白质组学方法, 从海岛棉7124根部蛋白中分离到一个IV型几丁质酶(GbCHI)。文章通过同源克隆获得了海岛棉GbCHI基因的cDNA序列, 并对该基因的表达特征及其蛋白的抑菌功能进行了分析鉴定。qRT-PCR实验结果表明GbCHI基因在棉花根、茎、叶、花和胚珠中均有表达, 其表达受大丽轮枝菌、水杨酸(SA)、乙烯(ACC)和茉莉酸(JA)诱导; 亚细胞定位分析显示GbCHI蛋白主要分布在细胞膜上; 体外抑菌实验证明GbCHI蛋白能显著抑制大丽轮枝菌孢子的萌发和菌丝的生长。这些研究结果为了解GbCHI的功能及其在抗黄萎病棉花分子育种中的应用提供了实验依据和思路。

本文引用格式

马银平,王付欣,杨淳淋,沈法富,夏桂先 . 海岛棉几丁质酶基因GbCHI的克隆与功能分析[J]. 遗传, 2012 , 34(2) : 240 -247 . DOI: 10.3724/SP.J.1005.2012.00240

Abstract

Chitinase is one of the important pathogenesis-related (PR) proteins in plants. By comparative proteomics study, a novel pathogen-responsive chitinase (known as GbCHI) has been identified from sea-island cotton (Gossypium barbadense). The GbCHI cDNA was cloned from wilt-resistant sea-island cotton and the anti-fungal activity of the gene product was investigated. qRT-PCR analysis indicated that GbCHI was expressed constitutively in root, stem, leaf, flower, and ovule of cotton plant, and the expression could be induced by Verticillium dahliae and plant hormone SA, ACC, and JA. Subcellular localization analysis using GFP-tagged proteins showed that GbCHI-GFP fusion proteins were targeted mainly to the plasma membrane. Anti-fungal assay demonstrated that GbCHI could inhibit spore germination and hyphae growth of V. dahliae significantly. These results provide important information for under-standing the cellular function of GbCHI and for exploring the application potential of this gene in molecular breeding of wilt-tolerant cotton plants.

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