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

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

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（S_13-bS_13-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BoSRK_13-b的激酶域存在相互作用。

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（S_13-bS_13-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 BoSRK_13-b.

关键词

羽衣甘蓝 / FERONIA / 自交不亲和性 / 表达分析 / 酵母双杂交

Key words

Brassica oleracea var. acephala / FERONIA / self-incompatibility / expression analysis / yeast two-hybrid

Author summay

荀宝茹（1999—），女，硕士研究生，主要从事植物发育研究。