甘蓝型油菜BnCNGC基因家族鉴定及其在核盘菌侵染和PEG处理下的表达特性分析  

Genome-wide identification of BnCNGC and the gene expression analysis in Brassica napus challenged with Sclerotinia sclerotiorum and PEG-simulated drought

作　　者：陈松余 丁一娟[1] 孙峻溟 黄登文 杨楠 代雨涵 万华方[1] 钱伟[1] CHEN Song-Yu;DING Yi-Juan;SUN Jun-Ming;HUANG Deng-Wen;YANG Nan;DAI Yu-Han;WAN Hua-Fang;QIAN Wei(College of Agronomy and Biotechnology,Southwest University/Academy of Agricultural Sciences,Southwest University,Chongqing 400716,China)

机构地区：[1]西南大学农学与生物科技学院/西南大学农业科学研究院,重庆400716

出　　处：《作物学报》2022年第6期1357-1371,共15页Acta Agronomica Sinica

基　　金：重庆市自然科学基金项目(cstc2020jcyj-msxmX0957);国家重点研发计划项目(2018YFD0100500)资助。

摘　　要：CNGC是植物中普遍存在的环核苷酸门控通道,对植物的逆境响应有重要作用。系统分析甘蓝型油菜BnCNGC基因家族成员全基因组分布、结构、进化及其响应不同逆境胁迫的表达特性,对于阐明其生物学功能具有重要意义。本研究利用拟南芥和甘蓝CNGC蛋白保守结构域及特异基序氨基酸序列在全基因组水平鉴定了甘蓝型油菜BnCNGC家族成员,分析其基因结构、染色体定位、蛋白理化性质、蛋白保守结构域、系统进化及启动子顺式作用元件等。利用转录组数据,筛选甘蓝型油菜逆境响应候选BnCNGC成员,并采用实时荧光定量PCR分析其在核盘菌、PEG模拟干旱胁迫下的表达模式。结果显示,共鉴定到49个甘蓝型油菜BnCNGC成员,分布于除A08、C06的17对染色体上,含有5~10个内含子,其上游1500 bp包含大量逆境胁迫响应元件。BnCNGC家族成员编码的蛋白质含413~801个氨基酸,相对分子量(MW)范围47.62~110.58kD,等电点(pI)范围6.10~9.88。系统进化分析表明,BnCNGC分为Group Ⅰ、Ⅱ、Ⅲ和Ⅳ四类。转录组数据分析表明, BnCNGC9、BnCNGC27和BnCNGC48均参与逆境胁迫响应。实时荧光定量PCR分析表明,甘蓝型油菜叶片中BnCNGC9、BnCNGC27和BnCNGC48均在核盘菌胁迫下表达下调,下调量分别为48.2%~99.1%、79.4%~87.1%和39.7%~92.6%。PEG模拟干旱胁迫时,甘蓝油菜叶片中BnCNGC9在处理24 h后上调3.34倍, BnCNGC48在处理48 h后上调6.27倍, BnCNGC27对干旱胁迫不敏感。CNGCs are common cyclic nucleotide-gated channel proteins in plants, which play important roles in plant stress response. In order to elucidate the biological function of the BnCNGC genes in Brassica napus, we identified the BnCNGC family members in the genome and analyzed the distribution, structure, evolution, and the expression of candidate members in B. napus exposed to Sclerotinia sclerotiorum or PEG-simulated drought. In this study, employing CNGC amino acid sequences of Arabidopsis thaliana and Brassica oleracea as references, the homologies were acquired using BLASTP software in Brassica napus Genome Browser, and the gene family members were identified according to the conserved domain and specific motif of BnCNGC in plants. The gene structure, chromosome distribution, protein physicochemical property, subcellular localization,phylogenetic evolution, and promoter cis-acting regulatory elements were analyzed. The relative expression profiles of the candidate BnCNGC genes, screened out based on the transcriptome data in B. napus challenged with S. sclerotiorum or PEG-simulated drought, were analyzed by qRT-PCR. As a result, a total of 49 BnCNGC genes were identified in B. napus genome. They were distributed at 17 pairs of chromosomes, except for A08 and C06, and generally had 5–10 introns. The 1500 bp region in the upstream sequence of BnCNGC contained lots of cis-acting regulatory elements involved in stress response. Most of the corresponding proteins were composed of 413–801 amino acids with molecular weight(MW) of 47.62–110.58 kD and isoelectric point(p I) of 6.10–9.88. All the identified family members had specific motif of CNGC in the CNBD region. Phylogenetic analysis showed that BnCNGC were divided into four categories, namely Group Ⅰ, Ⅱ, Ⅲ, and Ⅳ. Transcriptome data analysis revealed that BnCNGC9, BnCNGC27, and BnCNGC48 were involved in stress response. The relative expression levels of the three genes were detected in B. napus leaves exposed to different stresses. All were down-reg

关 键 词：甘蓝型油菜 CNGC基因家族 核盘菌 PEG模拟干旱 表达分析 

分 类 号：S565.4[农业科学—作物学]