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

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-b)S_(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2682 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。

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 selfincompatibility line(S_(13-b)S_(13-b))of ornamental kale by RT-PCR,and the cloned cDNA sequence was 2682 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 Slocus receptor kinase BoSRK_(13-b).