陆地棉CLE基因家族的鉴定及GhCLE13参与调控棉花抗旱性的功能分析  

Identification of the CLE gene family in Gossypium hirsutum and functional analysis of the drought resistance of GhCLE13

作　　者：戎宇轩 惠留洋 王沛琦 孙思敏 张献龙[1] 袁道军[1] 杨细燕[1] RONG Yu-Xuan;HUI Liu-Yang;WANG Pei-Qi;SUN Si-Min;ZHANG Xian-Long;YUAN Dao-Jun;YANG Xi-Yan(National Key Laboratory of Crop Genetic Improvement/Hubei Hongshan Laboratory,Huazhong Agricultural University,Wuhan 430070,Hubei,China)

机构地区：[1]华中农业大学作物遗传改良全国重点实验室/湖北洪山实验室,湖北武汉430070

出　　处：《作物学报》2024年第12期2925-2939,共15页Acta Agronomica Sinica

基　　金：兵团财政科技计划项目(2022DB012);农业生物育种国家科技重大专项(2023ZD04038)资助。

摘　　要：CLAVATA3/Embryo surrounding region-related(CLE)小肽家族是植物中最大的小肽激素家族,在植物中广泛存在,且参与植物多个重要的生命活动。本研究对陆地棉CLE基因家族进行了鉴定,并对GhCLE基因家族成员进行基因结构、启动子顺式作用元件、蛋白质理化性质、系统发育的分析;通过RNA-seq数据构建了GhCLE基因家族成员在陆地棉各组织的表达谱及干旱处理下的表达模式,并筛选在棉花根系特异表达且受干旱诱导的CLE基因;最后通过VIGS技术对筛选出的GhCLE基因进行了抗旱功能的验证。结果显示,在陆地棉全基因组共鉴定出40个GhCLE基因,GhCLE基因结构较为简单,32个GhCLE基因没有内含子,所有基因编码的蛋白序列均包含12 aa的CLE结构域;GhCLE基因启动子区域包含多种光诱导响应、胁迫响应、激素响应和发育相关的顺式作用元件;GhCLE基因在陆地棉多个组织均有表达,筛选出了1个根系特异表达且受干旱诱导的GhCLE13-D-2基因。通过VIGS技术和MDA含量的测定和比较验证了GhCLE13-D-2基因提高棉花的抗旱性的功能。本研究为CLE小肽在植物抗逆方面的深入研究以及棉花种质创新提供了新的理论依据。The CLAVATA3/Embryo surrounding region-related(CLE)peptide family is the largest family of small peptide hormones in plants.It is widely present in various plant species and participates in multiple important life activities of plants.In this study,the CLE gene family was identified at the genome-wide level in Gossypium hirsutum,and their gene structures,cis-acting elements in the promoter,protein physicochemical properties,and phylogenetic analysis of GhCLE gene family members were analyzed.The expression profiles of GhCLE gene family members in various tissues were constructed using RNA-seq data.Finally,the drought resistance function of the screened GhCLE genes was validated by virus-induced gene silencing(VIGS)technology.The results showed that a total of 40 GhCLE genes were identified in the whole genome of Gossypium hirsutum.The structures of GhCLE genes were relatively simple,with 32 GhCLE genes having no introns,and their protein sequences all contained a 12 aa CLE domain.The GhCLE gene promoter region contains a variety of cis-acting elements related to light response,stress response,hormone response,and development.The gene expression profile showed that the GhCLE gene was expressed in multiple tissues of Gossypium hirsutum,with GhCLE13-D-2 being specifically expressed in root tissue and induced by drought.The function of the GhCLE13-D-2 gene in improving cotton drought resistance was validated through VIGS technology and the measurement and comparison of MDA content.This study provides a new theoretical basis for the in-depth study of small peptides,such as CLE,in plant drought resistance and cotton germplasm innovation.

关 键 词：陆地棉 CLE小肽基因家族 根系特异表达 抗旱 GhCLE13-D-2 

分 类 号：S562[农业科学—作物学]