Xinjiang Agricultural Sciences ›› 2025, Vol. 62 ›› Issue (1): 137-145.DOI: 10.6048/j.issn.1001-4330.2025.01.017

• Cultivation Physiology·Physiology and Biochemistry·Germplasm Resources·Molecular Genetics·Soil Fertilizer • Previous Articles     Next Articles

Functional analysis of a C2H2 zinc protein GhZFP8 gene in Gossypium hirsutum resistance to Verticillium wilt

CHENG Guanfu(), LU Guoqiang, CUI Yongxiang, HOU Aocheng, ZHANG Guoshuai, LIANG Chunyan, LEI Jianfeng, LU Wei, DAI Peihong, LI yue()   

  1. Xinjiang Key Laboratory for Ecological Adaptation and Evolution of Extreme Environment Biology/College of Life Sciences, Xinjiang Agricultural University, Urumqi 830052, China
  • Received:2024-08-11 Online:2025-01-20 Published:2025-03-11
  • Correspondence author: LI yue
  • Supported by:
    National Natural Science Foundation of China(32160494);Natural Science Foundation of Xinjiang Uygur Autonomous Region(2023D01E03);National Innovation and Entrepreneurship Training Program for College Students(202210758001);Key Scientific R & D Program Project of Xinjiang Uygur Autonomous Region(2022B02033-1)

陆地棉锌指蛋白基因GhZFP8黄萎病的抗性功能验证

程贯富(), 卢国强, 崔永祥, 侯奥成, 张国帅, 梁春燕, 雷建峰, 路伟, 代培红, 李月()   

  1. 新疆农业大学生命科学学院/新疆极端环境生物生态适应与进化重点实验室,乌鲁木齐 830052
  • 通讯作者: 李月
  • 作者简介:程贯富(1998-),男,山东济南人,硕士研究生,研究方向为棉花逆境分子生物学,(E-mail)cgfyouxi@163.com
  • 基金资助:
    国家自然科学基金项目(32160494);新疆维吾尔自治区自然科学基金项目(2023D01E03);国家级大学生创新创业训练计划项目(202210758001);新疆维吾尔自治区重点研发专项(2022B02033-1)

Abstract:

【Objective】 To explore the function of the terrestrial cotton C2H2-type zinc finger protein gene GhZFP8 in the response to Verticillium wilt in cotton in the hope of laying a theoretical foundation for mining cotton disease resistance genes. 【Methods】 The cotton gene GhZFP8 was obtained by homologous cloning and its biological function explored. Bioinformatics methods were used to analyze the physicochemical properties of the gene, while the VIGS vector for the gene was constructed and transformed into cotton by Agrobacterium-mediated method to preliminarily test the disease resistance function of GhZFP8. 【Results】 The open reading frame (ORF) of GhZFP8 was 789 bp, encoded as a hydrophilic protein containing 262 amino acids, a relative molecular weight was 28.12 kD, a basic amino acid with an isoelectric point of 8.16, a lipid index of 59.31, and an average hydrophobicity of-0.718. The absence of a signal peptide in the GhZFP8 protein predicted that GhZFP8 was a non-transmembrane protein. GhZFP8 protein revealed that it had 2 ZnF-C2H2 structural domains. Its secondary structure prediction revealed that the GhZFP8 protein contained 6 α-helices and 4 β-folds. GhZFP8 gene silenced plants showed relatively reduced resistance to Verticillium wilt. 【Conclusion】 The GhZFP8 gene plays a positive regulatory role in cotton resistance to verticillium wilt.

Key words: Verticillium wilt; GhZFP8; gene cloning; VIGS

摘要:

【目的】 探究陆地棉C2H2型锌指蛋白基因GhZFP8在棉花抗黄萎病反应中的功能,为挖掘棉花抗病基因奠定理论基础。【方法】 通过同源克隆方法得到一个陆地棉C2H2型锌指蛋白基因GhZFP8,利用生物信息学方法分析该基因的理化性质,构建该基因病毒诱导的基因沉默(VIGS)载体,通过农杆菌介导法转化棉花,检验GhZFP8的抗病功能。【结果】 GhZFP8开放阅读框(ORF)为789 bp、编码为262个氨基酸,相对分子量为28.12 kD、等电点为8.16、脂肪指数为59.31和平均疏水性为-0.718,为亲水性、碱性的非跨膜蛋白。GhZFP8蛋白具有2个ZnF-C2H2结构域,含有6个α-螺旋和4个β-折叠,属于C2H2型锌指蛋白。GhZFP8基因沉默植株对黄萎病菌的抗性相对减弱。【结论】 GhZFP8基因在棉花抗黄萎病过程中发挥正调控作用。

关键词: 黄萎病, GhZAT8, 基因克隆, 病毒诱导的基因沉默

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