陆地棉CRK基因家族抗黄萎病功能分析  

Functional analysis of the CRK gene family in Gossypium hirsutumresistance to Verticillium wilt

作　　者：裴洪伟 索佳慧 武海娜[1] 孙正文 张艳[1] 杨君[1] PEI Hongwei;SUO Jiahui;WU Haina;SUN Zhengwen;ZHANG Yan;YANG Jun(Collaborative Innovation Center for Cotton Industry of Hebei Province,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]河北农业大学河北省棉花产业协同创新中心,河北保定071001

出　　处：《北方农业学报》2024年第6期1-12,共12页Journal of Northern Agriculture

基　　金：国家重点研发计划项目(2022YFF1001403);河北省自然科学基金项目(C2022204001);河北省现代农业产业技术体系专项资金(HBCT2024100201);河北农业大学大学生创新创业训练基金项目(2022196)。

摘　　要：【目的】鉴定分析陆地棉(Gossypium hirsutum)富含半胱氨酸类受体激酶(CRK)基因家族成员,为棉花抗黄萎病育种提供基因资源和理论依据。【方法】根据全基因组测序结果,利用生物信息学技术在陆地棉农大棉8号基因组中鉴定GhCRK基因家族,并进行蛋白理化性质、染色体定位和遗传进化分析。基于基因表达数据挖掘潜在的黄萎病抗性基因,并通过病毒诱导的基因沉默(virus induced gene silencing,VIGS)技术对候选基因进行抗病功能鉴定。【结果】在陆地棉农大棉8号基因组中共鉴定到50个GhCRKs,分布在14条染色体上,编码蛋白长度为390~1 774个氨基酸残基,分子量为43.15~196.66 kDa,平均等电点为6.94,大多定位在细胞质膜。蛋白结构域分析显示,90%的GhCRKs(45个)含有2个Stress-antifung结构域。在GhCRKs启动子区鉴定到植物激素顺式响应元件茉莉酸甲酯、脱落酸、水杨酸等7类。筛选出14个受大丽轮枝菌诱导发生显著上调表达的GhCRKs,且GhCRK08和GhCRK33沉默后,陆地棉黄萎病症状更严重、病情指数显著增高。【结论】陆地棉农大棉8号基因组中存在大量的CRKs,其中14个为潜在重要的黄萎病抗性候选基因,进一步明确了GhCRK08和GhCRK33的抗病功能。【Objective】To identify and analyze the cysteine-rich receptor-like kinase(CRK)gene family members of Gossypium hirsutum,and provide genetic resources and theoretical basis for cotton Verticillium wilt resistance breeding.【Methods】According to the results of whole genome sequencing,the GhCRK gene family in the Gossypium hirsutum Nongdamian 8 genome was identified using bioinformatics methodologies,analyses were conducted on protein physicochemical properties,chromosomal localization and genetic evolution.Furthermore,gene expression data were employed to potential Verticillium wilt resistance genes,and the resistance function of candidate gene was assessed through virus induced gene silencing(VIGS)technology.【Results】The genome of Gossypium hirsutum cultivar Nongdamian 8 harbored a total of fifty GhCRKs,distributed across fourteen chromosomes.These length of the encoded protein was 390 to 1774 amino acid residues,resulting in diverse molecular weights spanning from 43.15 kDa to 196.66 kDa,the average isoelectric point was observed at 6.94.The majority of GhCRKs were predominantly localized in the plasma membrane.Protein domain analysis showed that 90%of GhCRKs(45)contained two Stress-antifung domains.The promoter regions of GhCRKs encompassed seven cis-acting elements relating to responsiveness to plant hormones,including methyl jasmonate,abscisic acid,salicylic acid,et al. Fourteen GhCRKs exhibited up regulation expression in response to Verticillium dahliae infection. Following the silencing of GhCRK08 and GhCRK33,Gossypium hirsutum exhibited exacerbated symptoms of Verticillium wilt and a significantly higher disease index.【Conclusion】The genome of Gossypium hirsutum Nongdamian 8 contains a substantial number of CRKs,among which 14 hold potential as significant candidate genes for Verticillium wilt resistance. Additionally,the role of GhCRK08 and GhCRK33 in Verticillium wilt resistance has been further elucidated.

关 键 词：陆地棉 富含半胱氨酸类受体激酶(CRK)基因家族 黄萎病 生物信息学 基因功能 基因表达 

分 类 号：S562[农业科学—作物学] Q786[生物学—分子生物学]