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

doi:10.7525/j.issn.1673-5102.2024.02.015

Abstract

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.

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

CLC Number:

S635

Baoru XUN, Hongtao QIN, Rui MA, Nanfeng GUO, Yunping LIU, Ying WU, Xingguo LAN. Gene Cloning, Expression and Interaction Protein Analysis of FERONIA in Brassica oleracea var. acephala[J]. Bulletin of Botanical Research, 2024, 44(2): 298-306.

Figures/Tables 9

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引物名 Primer name	引物序列（5′→3′） Primer sequence（5′→3′）	用途 Application
BoFER-CDS-F	ATGACGTTCACGGAGGGACG	基因克隆 Gene clone
BoFER-CDS-R	CTAACGTCCCTTAGGGTTCA
BoFER-KD-F	GCGTACCGGAGACGGAAGAG
BoFER-KD-R	CTAACGTCCCTTAGGGTTCA
qRT-PCR-BoFER-F1	TCCGCCAACATCAATAAAATTAACC	qRT-PCR
qRT-PCR-BoFER-R1	GGCTTGGTGATTGAGTTAGGATG	qRT-PCR
BoACTIN-F	GCAGACCGTA TGAGCAAAGA	内参基因 Reference gene
BoACTIN-R	GGAGGTGCAACCACCTTAAT	内参基因 Reference gene
PGADT7-FER-KD-F	CAGATTACGCTCATATGGCGTACCGGAGACGGAAGAG	酵母载体构建 Yeast vector construction
PGADT7-FER-KD-R	CGAGCTCGATGGATCCCTAACGTCCCTTAGGGTTCA	酵母载体构建 Yeast vector construction

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