甘蓝型油菜REVEILLE家族鉴定及诱导表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2023-1153

摘要/Abstract

摘要：

【目的】 REVEILLE家族基因具有重要生物学功能，而在甘蓝型油菜（Brassica napus L.）中该家族基因研究较少，深入认识甘蓝型油菜中RVE家族基因功能及其与非生物胁迫的关系。【方法】 利用生物信息学方法，在甘蓝型油菜、白菜（Brassica rapa）和甘蓝（B. oleracea）中分别鉴定到33、16和16个RVE基因，并对其系统进化、染色体定位、基因结构和理化性质以及启动子顺式元件进行分析。【结果】 所有的RVE蛋白被分为两个亚家族。33个BnaRVE分别定位在18条A亚基因组的染色体和15条C亚基因组的染色体上。大部分成员属于较稳定的疏水蛋白；多数Ka/Ks值小于1，说明该家族受到强烈的纯化选择作用。基因结构变异较大，内含子数目在4-11之间。共线性分析结果表明，甘蓝型油菜和拟南芥、白菜、甘蓝存在大量的同源基因。大部分甘蓝型油菜RVE家族成员在子叶和叶都有表达且表达量最多。BnaRVE启动子上含有大量与激素和生物逆境相关的元件，通过定量PCR分析，4个BnaRVE在ABA与MeJA诱导和低温胁迫下的表达量显著上调。【结论】 在甘蓝型油菜基因组中鉴定出33个BnaRVE家族成员。不同基因在不同发育时期和不同组织中表现出不同的表达模式。不同基因对各种胁迫存在响应，且表达模式不一。RVE家族成员对ABA、MeJA和冷胁迫具有正向响应功能。

关键词: 甘蓝型油菜, RVE基因家族, 生物信息学分析, 逆境, 植物激素, 表达分析, 表达谱

Abstract:

【Objective】 REVEILLE family genes have important biological functions, but there are few studies on this family gene in Brassica napus L. This project was conducted to further understand the function of RVE family genes in B. napus and their relationship with abiotic stress. 【Method】 Bioinformatics methods were used to identify33, 16 and 16 RVE genes in B. napus, B. rapa and B. oleracea, and to analyze their phylogenetic development, chromosome localization, gene structure, physical and chemical properties, and promoter cis-acting elements. 【Result】 All RVE proteins were classified into two subfamilies. The 33 BnaRVE were located on 18 chromosomes of the A sub-genome and 15 chromosomes of the C sub-genome. Most members belonged to relatively stable hydrophobic proteins. Most of the Ka/Ks values were < 1, indicating that the family was subjected to strong purification selection. The gene structure varied greatly, and the number of introns ranged from 4 to 11. Collinearity analysis showed that there were large number of homologous genes in B. napus and Arabidopsis, B. rapa and B. oleracea. Most RVE family members of B. napus expressed RVE in the cotyledon and leaves, and the expression was the highest. BnaRVE promoter contained a large number of cis-acting elements related to hormone and biological stress. Through quantitative PCR analysis, the expressions of 4 BnaRVE genes were significantly upregulated under ABA and MeJA induction as well as low temperature stress. 【Conclusion】 Thirty-three members of the BnaRVE family were identified in the genome of B. napus. Different genes showed different expression patterns in different developmental stages and different tissues. Different genes have different expression patterns in response to various stresses. RVE family members have positive response function to ABA, MeJA and cold stress.

Key words: Brassica napus L., RVE genes family, bioinformatics analysis, adverse stress, phytohormone, expression analysis, expression profile

刘蓉, 田闵玉, 李光泽, 谭成方, 阮颖, 刘春林. 甘蓝型油菜REVEILLE家族鉴定及诱导表达分析[J]. 生物技术通报, 2024, 40(6): 161-171.

LIU Rong, TIAN Min-yu, LI Guang-ze, TAN Cheng-fang, RUAN Ying, LIU Chun-lin. Identification and Induced-expression Analysis of REVEILLE Family in Brassica napus L.[J]. Biotechnology Bulletin, 2024, 40(6): 161-171.

图/表 8

表1 甘蓝型油菜RVE基因家族表达分析的实时荧光定量引物

Table 1 Real-time fluorescent quantitative primers for RVE gene family expression analysis in B. napus

Gene name	Forword primer sequence（5'-3'）	Reverse primer sequence（5'-3'）
BnaRVE1d	TGTCTCCTGTTTCCTTTGCATCACC	AACTCTCTCTGTTTACGAGTCTCTCTCT
BnaRVE8b	TTCGACCTCCGGTATGGGAAG	GAATTACCTTGAAGCACCGGAGG
BnaRVE3c	GTGTGGCTCTTACAACTTCAAACGCAT	CACCCAATTCCTCTTTGATCACTGGAA
BnaRVE5b BnaActin2	TCTCCACTTATTCGATCGAGATTGGAAG GTCTTCTCTGCTCTTCTCA	GTTCGTTAGCACCGCTCTTCTG CCATTCCAGTTCCATTGTC

图1 拟南芥、甘蓝型油菜、白菜和甘蓝RVE家族成员系统发育进化树

Fig. 1 Phylogenetic analysis of RVE family in A. thaliana, B. napus, B. rapa and B. oleracea

图2 甘蓝型油菜RVE家族基因染色体定位分布图

Fig. 2 Chromosomal location of the RVE family gene in B. napus

图3 甘蓝型油菜RVE家族成员保守基序、结构域及基因结构

Fig. 3 Conserved motif, domain and gene structure of RVE family members in B. napus

图4 甘蓝型油菜RVE家族成员启动子区域结合的顺式作用元件（A）及前10位CRE在基因组上的分布（B）

Fig. 4 Genomic distribution of cis-acting elements binding to promoter region of RVE family members in B. napus（A）and the top 10 CRE（B）in genome

图5 甘蓝型油菜RVE家族种内共线性分析（A）及RVE家族基因在甘蓝型油菜、拟南芥、甘蓝和白菜之间的共线性分析（B-D）

Fig. 5 Intra-specic collinearity analysis of B. napus RVE family（A）and collinearity analysis of RVE family genes among A. thaliana, B. napus, B. rapa and B. oleracea（B-D）

图6 甘蓝型油菜RVE家族基因表达分析

Fig. 6 Gene expression analysis of RVE family in B. napus

图7 BnaRVE在不同胁迫处理下的RT-qPCR表达分析 A：BnaRVE在MeJA处理中的表达；B：BnaRVE在ABA处理中的表达；C：BnaRVE在冷处理中的表达

Fig. 7 RT-qPCR expression analysis of BnaRVE under different stress treatments A: Expression of BnaRVE in MeJA processing.B: Expression of BnaRVE in ABA processing. C: Expression of BnaRVE in cold treatment

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