| 棉花GhPBL1基因克隆及抗枯萎病功能研究 |
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| 引用本文： | 赵曾强,李艳军,彭建智,朱金成,张薇.棉花GhPBL1基因克隆及抗枯萎病功能研究[J].南方农业学报,2022,53(12):3307-3317. |
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| 作者姓名： | 赵曾强 李艳军 彭建智 朱金成 张薇 |
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| 作者单位： | 1 石河子大学农学院, 新疆石河子 832000;2 新疆农垦科学院棉花研究所, 新疆石河子 832000 |
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| 基金项目： | 国家自然科学地区科学基金项目（32160476） |
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| 摘 要： | 【目的】克隆棉花中类受体胞质激酶（RLCK）基因GhPBL1，并进行抗枯萎病功能研究，为棉花抗病分子育种提供新的基因资源。【方法】基于枯萎病菌诱导棉花根部基因表达谱数据，并结合棉花数据库，从中筛选并克隆棉花RLCK家族基因GhPBL1，利用生物信息学软件分析其序列特征，采用实时荧光定量技术（qRT-PCR）检测基因在不同抗性品种的不同组织及枯萎病菌和外源激素处理下的表达情况，并通过病毒诱导基因沉默（VIGS）技术研究GhPBL1基因的抗枯萎病功能。【结果】GhPBL1基因的开放阅读框（ORF）为1116 bp，编码371个氨基酸残基，含有STYKc保守结构域，位于第81~360位氨基酸，属于RLCK家族基因。GhPBL1基因在抗病材料和感病材料的根、茎和叶中均有表达，且均在根中的相对表达量最高。枯萎病菌处理后，抗病材料中GhPBL1基因的表达水平明显高于感病材料。水杨酸（SA）和茉莉酸甲酯（MeJA）均能诱导GhPBL1基因的表达，但该基因对SA胁迫响应时间早于MeJA胁迫。与对照植株（TRV:00）相比，TRV:GhPBL1沉默植株更易感病，且其抗病相关基因GhEDS1、GhCCR1、GhHCT1和Gh4CL的相对表达量显著（P<0.05）较低，但GhPR5基因的相对表达量极显著（P<0.01）较高。【结论】 GhPBL1基因具有明显的组织表达特异性，在抗病材料和感病材料中的表达水平存在差异，能被枯萎病菌诱导表达上调，且能响应SA和MeJA胁迫，推测GhPBL1基因在棉花抗枯萎病和外源激素胁迫中发挥正调控作用。
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| 关 键 词： | 棉花 枯萎病菌 类受体胞质激酶 GhPBL1 表达分析 功能研究 |
| 收稿时间： | 2022-06-07 |
Cloning of GhPBL1 gene in cotton and its resistance function against Fusarium wilt |
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| ZHAO Zeng-qiang,LI Yan-jun,PENG Jian-zhi,ZHU Jin-cheng,ZHANG Wei.Cloning of GhPBL1 gene in cotton and its resistance function against Fusarium wilt[J].Journal of Southern Agriculture,2022,53(12):3307-3317. |
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| Authors: | ZHAO Zeng-qiang LI Yan-jun PENG Jian-zhi ZHU Jin-cheng ZHANG Wei |
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| Affiliation: | 1 College of Agronomy, Shihezi University, Shihezi, Xinjiang 832000, China;2 Cotton Research Institute, Xinjiang Academy of Agricultural Reclamation Sciences, Shihezi, Xinjiang 832000, China |
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| Abstract: | 【Objective】 GhPBL1, a receptor-like cytoplasmic kinase(RLCK) gene from cotton, was cloned and its resistance to Fusarium wilt was studied, which provided new gene resources for cotton disease-resistant molecular breeding.【Method】 Using the gene expression profile data of cotton roots induced by Fusarium oxysporum and combined with the cotton database, the RLCK family gene GhPBL1 was screened and obtained. Bioinformatics software was used to analyze the sequence characteristics of GhPBL1 gene, and real-time quantitative fluorescence(qRT-PCR) was used to detect the gene expression in different tissues of different resistant varieties, F. oxysporum and exogenous hormone treatments. The anti-Fusarium wilt function of GhPBL1 gene was studied by virus-induced gene silencing(VIGS) technology.【Result】 The open reading frame(ORF) of GhPBL1 gene was 1116 bp. It encoded 371 amino acids residues, contained a STYKc conserved domain located in 81-360 amino acids. It belonged to RLCK family gene. Expression analysis showed that the gene was expressed in roots, stems and leaves of resistant and susceptible materials, but the expression in roots was the highest. After F. oxysporum treatment, the expression level of GhPBL1 gene in resistant materials was higher than that in susceptible materials. Both salicylic acid(SA) and methyl jasmonate(MeJA) could induce the expression of GhPBL1 gene, but the response time of the gene to SA stress was earlier than MeJA stress. Compared with control plants(TRV:00), TRV:GhPBL1-silenced plants were more susceptible to disease, and the relative expression levels of resistance related genes GhEDS1, GhCCR1, GhHCT1 and Gh4CL were significantly lower than the control(P<0.05), but the relative expression levels of GhPR5 were extremely significantly higher than the control(P<0.01).【Conclusion】 The expression of GhPBL1 differs among the resistant and susceptible materials and has obvious tissue specificity. The gene is up-regulated after F. oxysporum treatment and is able to respond to SA and MeJA stress. It is speculated that GhPBL1 gene plays a positive regulatory role in cotton resistance to Fusarium wilt and exogenous hormone stress. |
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