海岛棉耐盐相关基因GbPLA1-32的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of the Salt Tolerance Related Gene GbPLA1-32 in Gossypium barbadense

作　　者：张红 张悦新 徐楠 芮存 范亚朋 王晶 韩明格 王琴琴 孙亮庆 陈修贵[2] 陆许可[2] 王俊娟[2] 王德龙[2] 王帅[2] 郭丽雪[2] 赵兰杰[2] 陈超[2] 陈全家 叶武威 Zhang Hong;Zhang Yuexin;Xu Nan;Rui Cun;Fan Yapeng;Wang Jing;Han Mingge;Wang Qinqin;Sun Liangqing;Chen Xiugui;Lu Xuke;Wang Junjuan;Wang Delong;Wang Shuai;Guo Lixue;Zhao Lanjie;Chen Chao;Chen Quanjia;Ye Wuwei(Engineering Research Centre of Cotton(Ministry of Education),College of Agriculture,Xinjiang Agricultural University,Urumqi,830052;State Key Laboratory of Cotton Biology,Research Base,Zhengzhou University,Key Laboratory for Cotton Genetic Improvement,MOA,Institute of Cotton Research of Chinese Academy of Agricultural Sciences,Anyang,455000)

机构地区：[1]新疆农业大学农学院,教育部棉花工程研究中心,乌鲁木齐830052 [2]中国农业科学院棉花研究所,棉花生物学国家重点实验室,郑州大学研究基地,农业部棉花遗传改良重点开放实验室,安阳455000

出　　处：《分子植物育种》2024年第5期1421-1429,共9页Molecular Plant Breeding

基　　金：国家自然科学基金青年基金项目(32001460)资助。

摘　　要：为了解海岛棉GbPLA1-32基因结构特征及功能,本研究以海岛棉‘海7124’为实验材料,利用前期耐盐转录组测序结果筛选得到差异表达基因并进行克隆。通过生物信息学分析,获得‘海7124’GbPLA1-32基因全长CDS,分析该基因编码蛋白的理化性质、鉴定其亲水性与疏水性,对磷酸化和糖基化位点、信号肽和跨膜区以及蛋白质结构进行预测,确定亚细胞定位并构建系统进化树。结果表明,该基因编码区全长1530 bp,编码509个氨基酸,编码蛋白在磷脂酶A1家族,属α/β-水解酶超家族。它属于亲水性蛋白,不存在明显信号肽且不具有跨膜结构,亚细胞定位于叶绿体中。同时,α-螺旋、延伸链、β-转角和无规则卷曲这4个元件构成了Gb PLA1-32蛋白的二级结构。进化树分析表明海岛棉与陆地棉亲缘关系最为接近,根据其编码蛋白将该基因命名为GbPLA1-32。RT-qPCR分析结果表明,GbPLA1-32基因能够对PEG和盐两种胁迫做出响应。本研究结果可为进一步阐明GbPLA1-32在海岛棉中的生物学功能提供参考。In order to understand the structural characteristics and functions of the sea island cotton GbPLA1-32 gene.In this study,the'Hai7124'in the sea island cotton was used as the experimental material,and the differentially expressed genes were screened and cloned using the results of salt-tolerant transcriptome sequencing.Through bioinformatics analysis,the full-length CDS of'Hai7124'GbPLA1-32 gene was obtained.Analyze the physical and chemical properties of the protein encoded by the gene.Identify its hydrophilicity and hydrophobicity.Prediction of phosphorylation and glycosylation sites,signal peptides and transmembrane regions,and protein structure.Determine subcellular location and construct a phylogenetic tree.The results showed that the coding re gion of the gene was 1530 bp in length,encoding 509 amino acids,and the encoded protein was in the phospholipase A1 family,belonging to theα/β-hydrolase superfamily.It is a hydrophilic protein,there is no obvious signal peptide and no transmembrane structure,subcellular localization in the chloroplast.At the same time,the four elements ofα-helix,extended strand,β-turn and random coil constitute the secondary structure of the GbPLA1-32 protein.The phylogenetic tree analysis showed that the relationship between sea island cotton and upland cotton is the closest.The gene was named GbPLA1-32 according to its coding protein.The results of RT-qPCR analysis showed that the GbPLA1-32 gene can respond to salt and alkali stress.The results of this study can provide references for further elucidating the biological functions of GbPLA1-32 in Gossypium barbadense.

关 键 词：海岛棉 磷脂酶A1 基因克隆 表达分析 生物信息学分析 

分 类 号：S562[农业科学—作物学]