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棉花学报 ›› 2021, Vol. 33 ›› Issue (6): 459-468.doi: 10.11963/cs20210061

• 研究与进展 • 上一篇    下一篇

陆地棉纤维起始期优势表达基因GhCRPK1的克隆及功能研究

王雪慧(),陈丽锦,赵若林,程海亮,张友平,王巧连,吕丽敏,宋国立,左东云*()   

  1. 中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南 安阳 455000
  • 收稿日期:2021-10-28 出版日期:2021-11-15 发布日期:2022-04-14
  • 通讯作者: 左东云 E-mail:1115193717@qq.com;zdy041@163.com
  • 作者简介:王雪慧(1993―),女,硕士研究生, 1115193717@qq.com
  • 基金资助:
    国家自然科学基金(31621005);中国农业科学院科技创新工程;中央级公益性科研院所基本科研业务费专项(1610162021013)

Cloning and functional analysis of GhCRPK1, a gene preferentially expressed during fiber initiation in upland cotton

Wang Xuehui(),Chen Lijin,Zhao Ruolin,Cheng Hailiang,Zhang Youping,Wang Qiaolian,Lü Limin,Song Guoli,Zuo Dongyun*()   

  1. Institute of Cotton Research, Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology, Anyang, Henan 455000, China
  • Received:2021-10-28 Online:2021-11-15 Published:2022-04-14
  • Contact: Zuo Dongyun E-mail:1115193717@qq.com;zdy041@163.com

摘要:

【目的】研究陆地棉类受体激酶GhCRPK1(Cold-responsive protein kinase 1,CRPK1)在棉花纤维发育中的功能。【方法】以陆地棉全基因组关联分析(Genome-wide association study,GWAS)数据及陆地棉纤维发育不同时期的表达数据,筛选到GhCRPK1在棉花纤维起始期优势表达,通过基因克隆、亚细胞定位等方法研究了GhCRPK1的功能,对过表达GhCRPK1的拟南芥及突变体crpk1的表皮毛长度和密度进行观察统计,利用酵母双杂交筛选与GhCRPK1相互作用的蛋白。【结果】GhCRPK1基因全长为4 594 bp,开放阅读框长度为1 047 bp,编码348个氨基酸,在开花当天到开花后3 d持续优势表达,编码产物定位在细胞膜上。GhCRPK1在拟南芥中异源表达后可促进表皮毛的伸长;拟南芥中同源基因突变后,表皮毛变短。酵母双杂交实验发现,GhCRPK1与热激蛋白Gh_D08G0057相互作用。【结论】GhCRPK1在棉花纤维发育起始时期优势表达,定位在细胞膜上,在拟南芥中异源表达后调控拟南芥表皮毛的伸长。

关键词: 类受体激酶; GhCRPK1; 纤维发育; 表皮毛; 信号传导

Abstract:

[Objective] The aim of this article is to study the function of GhCRPK1 (Cold-responsive protein kinase 1, CRPK1) during fiber development in upland cotton. [Method] According to the genome-wide association study (GWAS) data and the transcriptome data of fiber development at different stages in cotton, GhCRPK1 may be involved in fiber initiation by gene clone, subcellular localization to study the GhCRPK1. The full length of GhCRPK1 was amplified and transformed into Arabidopsis, and the length and density of trichomes of the GhCRPK1-overexpressed plants and Arabidopsis mutant crpk1 were observed. At last, yeast two-hybrid assay was performed to screen the interaction proteins. [Result] The full-length of GhCRPK1 is 4 594 bp containing a 1 047 bp open reading frame and encoding 348 amino acids. The expression level of GhCRPK1 peaked at the 0-3 days post anthesis. The GhCRPK1 was localized on the plasma membrane. The Arabidopsis plants overexpressing GhCRPK1 had longer trichomes compared with the wild type. While the trichomes were shorter in the crpk1 mutant. Yeast two-hybrid analysis revealed that GhCRPK1 can interact with a heat shock protein Gh_D08G0057. [Conclusion] GhCRPK1, a gene preferentially expressed during fiber initiation and localized on the plasma membrane, regulated the elongation of trichome in Arabidopsis.

Key words: receptor like kinase; GhCRPK1; fiber development; trichome; signal transduction