摘要
为明确茎瘤芥瘤状茎形成过程中的糖酸含量变化,揭示茎瘤芥甘油醛-3-磷酸脱氢酶(GAPDH)的主要生理功能,并进一步探究BjuGAPC基因的表达模式对糖酸含量的调控机制,首先对BjuGAPC基因进行克隆,同时,利用生物信息学方法对BjuGAPC的理化性质、系统进化发育等多方面进行系统分析,并使用qRT-PCR比较分析BjuGAPC基因在茎瘤芥茎不同发育阶段的表达规律。结果显示:(1)BjuGAPC基因开放阅读框全长1 038 bp,是一个稳定的亲水性蛋白质,同时具有多个蛋白质糖基化位点、磷酸化位点和特异性蛋白质激酶的结合位点;BjuGAPC与其他物种的GAPDH具有高度相似性,亲缘关系与同科的芜菁、油菜、萝卜较近。(2)qRT-PCR分析结果表明,BjuGAPC基因在茎瘤芥茎膨大不同时期的相对表达量差异显著且随着器官膨大相对表达量下调,与转录组的表达丰度一致。(3)相关性分析结果显示,基因表达量与可溶性糖和可滴定酸含量均呈正相关关系,并与糖酸比呈显著正相关关系。说明BjuGAPC基因参与甘油醛-3-磷酸脱氢酶中NAD+-GAPDH的合成,BjuGAPC基因对糖酸含量有正调控作用,可能参与了糖酸的合成过程。
The aim of the study was to clarify the changes of sugar and acid contents during the formation of tuberculate stem in Brassica juncea var. tumida and reveal the main physiological functions of glyceraldehyde-3-phosphate dehydrogenase(GAPDH), and to further explore the regulation mechanism of BjuGAPC gene expression mode on sugar and acid content. BjuGAPC gene was cloned first, and its physicochemical properties, system evolution development and other aspects were systematically analyzed by bioinformatics methods at the same time. qRT-PCR method was used to compare and analyze the expression pattern of BjuGAPC gene in different development stages of B. juncea var. tumida stem. The results showed that, firstly, the overall length of open reading frame of BjuGAPC gene was 1 038 bp, and it was a stable and hydrophilic protein with multiple glycosylation sites, phosphorylation sites and specific protein kinase binding sites. BjuGAPC had a high similarity with GAPDH of other species, and its genetic relationship was closely related to crops of the same family, such as Brassica rapa, Brassica napus and Raphanus sativus. Secondly, results of qRT-PCR analysis showed that, the relative expression level of BjuGAPC gene at different stages of B. juncea var. tumida stem swelling was significantly different and was down-regulated with the organ swelled, which was consistent with the expression abundance of transcriptome. Thirdly, correlation analysis revealed that, gene expression was positively correlated with soluble sugar content and titratable acid content, and was significantly positively correlated with sugar-acid ratio. The study revealed that, BjuGAPC gene participated in the synthesis of NAD+-GAPDH in GAPDH, and showed positive regulatory role in sugar and acid contents, which may be involved in the synthesis process of sugar and acid in Brassica juncea.
作者
李晓燕
李成双
金业程
魏小涵
李梦瑶
LI Xiao-yan;LI Cheng-shuang;JIN Ye-cheng;WEI Xiao-han;LI Meng-yao(College of Horticulture,Sichuan Agricultural University,Chengdu 611130,China)
出处
《江苏农业学报》
CSCD
北大核心
2022年第6期1474-1483,共10页
Jiangsu Journal of Agricultural Sciences
基金
国家自然科学基金项目(32002027)。
关键词
茎瘤芥
茎膨大
糖酸含量
BjuGAPC基因
理化性质
表达分析
Brassica juncea var.tumida
stem swelling
sugar and acid content
BjuGAPC gene
physical and chemical properties
expression analysis
