陆地棉GhCEF-G基因家族鉴定及在种子萌发中的功能分析  

Genome-wide identification of GhCEF-G gene family in Gossypium hirsutum and functional analysis in seed germination

作　　者：司爱君 陈斌[1] 王志城[1] 陈丽婷 张艳 王省芬[1] SI Aijun;CHEN Bin;WANG Zhicheng;CHEN Liting;ZHANGYan;WANG Xingfen(Key Laboratory for Crop Gennplasm Resources of Hebei/College of Agronomy,Hebei Agricultural University,Baoding 071000,China;Cotton Research Institute,Xinjiang Academy of Agricultural and Reclamation Science/Key Laboratory for Cotton Biology and Genetic Breeding of Northwestern Inland Region,Ministry of Agriculture,Shihezi 832000,China)

机构地区：[1]河北农业大学河北省作物种质资源重点实验室/农学院,河北保定071000 [2]新疆农垦科学院棉花研究所/农业部西北内陆棉区棉花生物学与遗传育种重点实验室,新疆石河子832000

出　　处：《河北农业大学学报》2023年第1期22-30,共9页Journal of Hebei Agricultural University

基　　金：国家棉花产业技术体系项目(CARS-15-03);河北省重点研发计划项目(21326314D)。

摘　　要：延伸因子G(EF-G)是蛋白质生物合成过程中肽链延长必需的调节因子之一,对叶绿体编码蛋白的翻译至关重要。本研究以陆地棉‘新陆早28号’基因组数据为参考,运用生物信息学对GhCEF-G家族成员的染色体定位、基因结构、保守结构域和顺式元件进行了分析,并基于前期关于种子萌发性状的GWAS分析,从陆地棉鉴定并克隆得到叶绿体延伸因子GhCEF-G06基因。结果显示,陆地棉GhCEF-G基因家族包含19个成员,其编码区由2~21个外显子组成,编码蛋白含有6个保守基序,保守性较强;GhCEF-G基因上游2 kb序列中存在激素及逆境胁迫响应等顺式作用元件。GhCEF-G06基因开放阅读框全长2352 bp,编码783个氨基酸,蛋白的分子量为86.31 kD,理论等电点为5.54,属亲水性氨基酸;转录组数据的基因表达量热图显示,GhCEF-G06基因在吸胀后种子的子叶和根中高表达;qRT-PCR分析表明,GhCEF-G06基因在‘新陆早28号’种子吸胀时期的表达量较高。进化关系分析表明,GhCEF-G06氨基酸序列与同为锦葵科的木槿氨基酸序列相似性最高,其次是锦葵目的榴莲与可可。GhCEF-G06基因过表达拟南芥的表型鉴定显示,转基因植株的萌发和幼苗生长速度比野生型慢;经夫西地酸(Fusidic acid,FA)处理后,转基因植株萌发恢复正常。陆地棉GhCEF-G基因家族成员的鉴定及功能分析为深入解析GhCEF-G调控棉花种子萌发的分子机制提供重要参考。Elongation factor G(EF-G) is one of the necessary regulators for peptide chain elongation during protein biosynthesis, which is crucial for the translation of chloroplast-encoded proteins. In this study, the chromosome localization, gene structure, conserved domains and cis-elements of GhCEF-G family members were analyzed via bioinformatics based on the genomic data of Gossypium hirsutum Nongdamian8. Then the chloroplast elongation factor GhCEF-G06 was identified and cloned from upland cotton for functional study according to our previous GWAS analysis of seed germination traits.The results showed that the GhCEF-G gene family contained 19 members,and the gene coding region was consisted of 2-21 exons.The GhCEF-G proteins had six highly conserved motifs.The 2 kb upstream sequences of GhCEF-G genes contained cis-acting elements of hormone and stress responses.The length of open reading frame of GhCEF-G06 was 2 352 bp encoding a hydrophilic protein with 783 amino acids. The molecular weight of GhCEF-G06 protein was 86.31 kD, and its theoretical isoelectric point was 5.54. The expression analysis using transcriptome data showed that GhCEF-G06 was highly expressed in the cotyledons and roots of imbibitional seeds. The qRT-PCR analysis showed that the expression of GhCEF-G06 was higher during the imbibition stage of ‘Xinluzao 28’ seed.Phylogenetic analysis showed that GhCEF-G06 protein sequence shared the highest similarity to homology in hibiscus that is also in the mallow family. It also similar to homologies in durian and cocoa in the mallow order. Phenotypic identification of GhCEF-G06 over expressed Arabidopsis showed that germination and seedling growth of transgenic plants were slower than wild-type plants. The seeds of the transgenic plants germinated normally as wild-type when treated with fusidic acid(FA).The identification and functional analysis of GhCEF-G provided a reference for further analysis of the molecular mechanism of GhCEF-G genes regulating cotton seed germination.

关 键 词：陆地棉 延伸因子 种子萌发 表达 

分 类 号：S188[农业科学—农业基础科学]