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紫色马铃薯bHLH转录因子家族基因的鉴定及表达分析

Identification and Expression Analysis of bHLH Transcription Factor Family Genes in Purple Potato

  • 摘要: bHLH转录因子家族是第二大转录因子家族,在植物生长发育、花青素生物合成途径的调控、逆境胁迫响应等发挥重要作用。为挖掘马铃薯(Solanum tuberosum)的紫色块茎花青素合成相关的bHLH基因,本研究基于紫色马铃薯转录组测序数据筛选bHLH基因,通过生物信息学方法对其进行序列分析,对bHLH基因在不同时期的表达与功能进行分析与预测,并通过RT-qPCR技术分析花青素合成相关的5个bHLH基因在不同发育时期的表达水平。结果表明:紫色马铃薯中筛选出103个bHLH基因,编码的蛋白大小在129~699 aa,等电点在4.65~9.75,均为亲水性蛋白;碱性区域由15个碱性氨基酸组成,均有高度保守的H5-E9-R12序列(His5-Glu9-Arg12),两个螺旋区内含有高度保守的亮氨酸(L)、精氨酸(R)和缬氨酸(V);基序预测得到2个保守基序,最长的基序含35个氨基酸;有36个bHLH基因在整个生育期呈下调表达、有67个bHLH基因呈上调表达,其中41个bHLH基因在3个发育时期差异表达显著;103个紫色马铃薯bHLH基因与拟南芥(Arabidopsis thaliana)bHLH基因聚类为13个亚族;GO富集分析发现103个bHLH基因广泛参与花青素生物合成、防御反应、生长发育等72项功能,其中参与花青素和类黄酮合成的bHLH基因有5个,其RT-qPCR结果与测序表达一致。

     

    Abstract: The bHLH transcription factor family was known as the second largest transcription factor family, which plays important roles in the process of plant growth and development, the regulation of anthocyanin biosynthesis pathway, adversity stress and etc. In order to discover the bHLH genes related to the synthesis of anthocyanins, bHLH genes were screened based on the transcriptome sequencing data of purple potato, and the sequence of bHLH were analyzed by bioinformatics method, and the expression and function of bHLH genes were analyzed and predicted at different stages. Then the expression levels of five bHLH genes related to anthocyanin synthesis at different developmental stages were analyzed by RT-qPCR. The results showed that 103 bHLH genes in purple potatoes were screened, and the sizes of encoded proteins size was ranged from 129 aa to 699 aa, and the isoelectric point was ranged from 4.65 to 9.75, belonging to hydrophilic proteins. There was a highly conserved H5-E9-R12 sequence(His5-Glu9-Arg12) in the basic region, which was composed of 15 basic amino acids. There were highly conserved leucine(L), arginine(R)and valine(V)in two helical regions. Two conservative motifs were predicted, and the longest motif contained 35 amino acids. In different developmental stages, 36 bHLH genes were down-regulated expression, and 67 bHLH genes were up-regulated expression trend, of which 41 bHLH genes were differentially expressed during the three developmental stages; 103 bHLH genes of purple potato were divided into 13 sub-families clustering with bHLH family Arabidopsis thaliana; GO enrichment analysis revealed that 103 bHLH genes were widely involved in 72 functions such as anthocyanin biosynthesis, defense response, growth and development, among which five bHLH genes might be involved in the synthesis of anthocyanins and flavonoids. The results of RT-qPCR were consistent with sequencing expression trends.

     

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