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植物研究 ›› 2024, Vol. 44 ›› Issue (2): 298-306.doi: 10.7525/j.issn.1673-5102.2024.02.015

• 分子生物学 • 上一篇    下一篇

羽衣甘蓝类受体激酶FERONIA基因克隆、表达及与相互作用蛋白分析

荀宝茹, 秦洪涛, 马蕊, 郭楠枫, 刘运平, 吴莹, 蓝兴国()   

  1. 东北盐碱植被恢复与重建教育部重点实验室,东北林业大学,哈尔滨 150040
  • 收稿日期:2023-11-29 出版日期:2024-03-20 发布日期:2024-03-11
  • 通讯作者: 蓝兴国 E-mail:lanxingguo@126.com
  • 作者简介:荀宝茹(1999—),女,硕士研究生,主要从事植物发育研究。
  • 基金资助:
    黑龙江省大学生创新创业训练计划项目(S202310225227);国家自然科学基金面上项目(31870300)

Gene Cloning, Expression and Interaction Protein Analysis of FERONIA in Brassica oleracea var. acephala

Baoru XUN, Hongtao QIN, Rui MA, Nanfeng GUO, Yunping LIU, Ying WU, Xingguo LAN()   

  1. Key Laboratory of Saline-alkali Vegetation Restoration and Reconstruction in Northeast China,Northeast Forestry University,Harbin 150040
  • Received:2023-11-29 Online:2024-03-20 Published:2024-03-11
  • Contact: Xingguo LAN E-mail:lanxingguo@126.com

摘要:

FERONIA(FER)受体激酶在有性生殖中的花粉与柱头信号识别中发挥重要作用,为分析类受体激酶FERONIA(FER)在羽衣甘蓝(Brassica oleracea var. acephala)授粉中的作用,以羽衣甘蓝自交不亲和系(S13-bS13-b )为研究材料,采用RT-PCR从柱头中克隆得到BoFER基因,获得的BoFER cDNA序列,全长2 682 bp编码893个氨基酸,其含有高度保守激酶结构域和Ser/Thr激酶结合位点。利用qRT-PCR技术对BoFER在授粉过程中的表达水平进行分析,结果发现BoFER在不亲和授粉过程其表达量逐渐升高,而在亲和授粉过程中其表达量逐渐降低。利用酵母双杂交分析BoFER与已知的自交不亲和相关蛋白的相互作用,结果显示BoFER与S位点受体激酶BoSRK13-b的激酶域存在相互作用。

关键词: 羽衣甘蓝, FERONIA, 自交不亲和性, 表达分析, 酵母双杂交

Abstract:

FERONIA (FER) receptor kinase plays an important role in pollen and stigma recognition in flowering plants. To analyze the function of receptor-like kinase FERONIA(FER) in the pollination in ornamental kale(Brassica oleracea var. acephala), BoFER gene was cloned from the stigma of the self-incompatibility line(S13-bS13-b ) of ornamental kale by RT-PCR, and the cloned cDNA sequence was 2 682 bp, which encoded 893 amino acids containing the highly conserved kinase domain and Ser/Thr kinase binding site, and the expression level of BoFER was analyzed by qRT-PCR. The results showed that the expression of BoFER in stigmas gradually increased after incompatible pollination, but gradually decreased after compatible pollination. Furthermore, the interaction between BoFER and known proteins related to self-incompatibility were analyzed by yeast two-hybrid. The results suggested that BoFER interacted with the kinase domain of S-locus receptor kinase BoSRK13-b.

Key words: Brassica oleracea var. acephala, FERONIA, self-incompatibility, expression analysis, yeast two-hybrid

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