棉花2个胚胎发育晚期富集蛋白基因的克隆与表达分析

doi:10.7668/hbnxb.20190096

华北农学报 ›› 2019, Vol. 34 ›› Issue (6): 33-38. doi: 10.7668/hbnxb.20190096

所属专题： 棉花；生物技术；

• 作物遗传育种·种质资源·生物技术 • 上一篇下一篇

棉花2个胚胎发育晚期富集蛋白基因的克隆与表达分析

甄军波¹, 刘迪¹, 宋世佳², 刘琳琳¹, 蔡肖¹, 张曦¹, 李海山², 迟吉娜¹

1. 河北省农林科学院棉花研究所, 农业部黄淮海半干旱区棉花生物学与遗传育种重点实验室, 河北石家庄 050051;
2. 河北省农林科学院, 河北石家庄 050051

收稿日期:2019-08-27 出版日期:2019-12-28
通讯作者: 迟吉娜(1977-),女,山东掖县人,研究员,博士,主要从事棉花功能基因组和资源创新研究;李海山(1980-),男,河北唐山人,副研究员,主要从事农业技术研究和人事管理工作。
作者简介:甄军波(1984-),男,河北无极人,助理研究员,硕士,主要从事棉花抗逆分子生物学研究。
基金资助:
河北省科技支撑计划子课题（16226303D-3）；河北省现代农业科技创新工程子课题（2019-3-7-3）；河北省棉花产业技术体系（HBCT2018040207）

Cloning and Expression Analysis of Two Late Embryogenesis Abundant(LEA) Proteins from Upland Cotton

ZHEN Junbo¹, LIU Di¹, SONG Shijia², LIU Linlin¹, CAI Xiao¹, ZHANG Xi¹, LI Haishan², CHI Ji¹

1. Cotton Research Institute, Hebei Academy of Agriculture and Forestry Sciences, Key Laboratory of Cotton Biology and Genetic Breeding in Huanghuaihai Semiarid Area, Ministry of Agriculture, Shijiazhuang 050051, China;
2. Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050051, China

Received:2019-08-27 Published:2019-12-28

摘要/Abstract

摘要： LEA基因以基因家族的形式在高等植物中广泛存在，在植物生长发育及逆境胁迫应答的过程中发挥重要的作用。为深入研究棉花LEA基因的作用和功能，利用RT-PCR的方法，从棉花纤维均一化cDNA文库中分离得到2个LEA家族基因，分别命名为GhLEA和GhLEAG1（登录号分别为KF906314和KF906316）。GhLEA全长948 bp，编码315个氨基酸，等电点4.4，分子质量为35 ku，GhLEAG1全长756 bp，编码251个氨基酸，等电点10.76，分子质量为27 ku。系统进化树表明，GhLEA属于LEA2亚家族，GhLEAG1属于LEA3亚家族，2个基因的保守基序组成有明显的差异。荧光实时定量PCR表明，GhLEA和GhLEAG1均在棉花纤维20DPA表达量最高，GhLEA在棉花根中表达量最高，而GhLEAG1在茎中的表达量最高。在高盐和PEG处理条件下，2个基因的表达模式差异明显，此外，2个基因还受到脱落酸（ABA）的诱导表达，基因表达量均呈现先升高后降低的趋势，推测2个基因均参与了棉花高盐、PEG等非生物胁迫应答。本研究可为进一步研究LEA蛋白在棉花中的功能提供理论依据。

关键词: 棉花, LEA, 非生物胁迫, 序列分析, 表达分析

Abstract: The LEA genes often exist as a large gene family in higher plants, and have the pivotal role in plant growth and development, and various stress response. To further study the function of cotton LEA genes, two LEA genes were isolated from a cotton fiber-normolized cDNA library, which were named GhLEA and GhLEAG1 (GenBank accession number KF906314 and KF906316). GhLEA contained an open reading frame(ORF) of 948 bp, encoding a protein of 315 amino acids, with a molecular weight of 35 ku, and a theoretical isoelectric point of 4.4. GhLEAG1 contained an open reading frame(ORF) of 756 bp, encoding a peptide of 251 amino acids, with a molecular weight of 27 ku, and a theoretical isoelectric point of 10.76. Phylogenetic tree showed that GhLEA and GhLEAG1 had different motif elements, belonging to LEA2 and LEA3, separately. Quantitative RT-PCR analysis showed that both GhLEA and GhLEAG1 had the expression peak at 20DPA, and the expression of GhLEA in root was the highest, but the expression of GhLEAG1 was the highest in stem. The expression patterns of the two genes were significantly different under salt stress and PEG. In addition, the two genes were also induced by ABA, the expression level was increased first and then decreased. The data indicated that GhLEA and GhLEAG1 might play an important role in the abiotic stress response of cotton. This study will provide an important reference for further research on the molecular function of LEA in cotton.

Key words: Cotton, LEA, Abiotic stress, Sequence analysis, Expression analysis

中图分类号:

Q78
S562.03

甄军波, 刘迪, 宋世佳, 刘琳琳, 蔡肖, 张曦, 李海山, 迟吉娜. 棉花2个胚胎发育晚期富集蛋白基因的克隆与表达分析[J]. 华北农学报, 2019, 34(6): 33-38. doi: 10.7668/hbnxb.20190096.

ZHEN Junbo, LIU Di, SONG Shijia, LIU Linlin, CAI Xiao, ZHANG Xi, LI Haishan, CHI Ji. Cloning and Expression Analysis of Two Late Embryogenesis Abundant(LEA) Proteins from Upland Cotton[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(6): 33-38. doi: 10.7668/hbnxb.20190096.

http://www.hbnxb.net/CN/Y2019/V34/I6/33

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Cloning and Expression Analysis of Two Late Embryogenesis Abundant(LEA) Proteins from Upland Cotton

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