紫背天葵叶片中花青素种类及合成调控基因转录组分析
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引用本文:张少平,邱珊莲,张 帅,洪佳敏,林宝妹,郑开斌.紫背天葵叶片中花青素种类及合成调控基因转录组分析[J].西北植物学报,2019,39(5):808~816
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张少平1,2,邱珊莲1,张 帅1,洪佳敏1,林宝妹1,郑开斌1,2* (1 福建省农业科学院 亚热带农业研究所福建漳州 3630052 福建省农业科学院 作物研究所福州350013) 
基金项目:福建省农业科学院创新团队(STIT2017 2 11);
中文摘要:为获取紫背天葵(Cynura bicolor DC.)叶片中花青素种类及其合成调控基因等信息,该试验以紫背天葵叶背面紫色以及经处理叶背面几乎全绿(对照)的叶片为材料,进行转录组测序分析,同时进行6个相关差异表达基因的qRT PCR分析和6种花青素苷元的HPLC检测,以揭示紫背天葵特有的花青素苷元及其合成调控关键基因信息。结果表明:(1)在紫背天葵中共获得14个花青素苷元及32个花青素合成调控基因信息,其中表达量差异显著下调的4个基因为 Pg(c11692)、Cy(c42112)、ANS(c38551)和3GT(c9064),表达量差异显著上调的2个基因是D FR(c35961)和3GT(c20283)。(2)qRT PCR检测结果显示,上述6个基因在2种紫背天葵叶中的表达趋势(上调或下调)与转录组测序结果完全一致,但转录组测序检测到的表达趋势差异倍数比qRT PCR检测结果更加明显。(3)HPLC分析显示,紫背天葵叶中均未检测到Dp、Pt、Pn及Mv等4类花青素苷元,但紫背天葵叶中富含Cy花青素苷元,且背面紫色的叶中Cy类花青素苷元含量(62.21 mg/kg)显著高于绿色叶对照(6.86 mg/kg);背面紫色和全绿叶中的Pg花青素苷元含量均低于0.43 mg/kg。研究推测,Cy和Pg花青素苷元在绿叶紫背天葵(对照)中含量显著降低可能是因为存在1个ANS和1个3GT正调控以及1个DFR和1个3GT负调控所致。
中文关键词:紫背天葵  转录组测序  花青素  花青素苷元  调控基因
 
Transcriptome Analysis of Anthocyanidins and Their Synthesis Regulatory Genes in Gynura bicolor Leaves
Abstract:In order to explore different anthocyanidins and their synthesis regulator genes in Gynura bicolor leaves, we sequenced the transcriptomes of 2 G. bicolors including the purple leaves and the green leaves in control by Illumina HiSeq 2500 platform. At the same time, previously reported 6 kinds of anthocyanidins were detected by HPLC and 6 differential expressed genes were identified by the RT PCR.The aim is to reveal its unique anthocyanidins and key gene information for its synthesis and regulation in G. bicolor. The results showed that: (1) 14 unigenes related to anthocyanidins and 34 unigenes related to their synthesis regulator genes were obtained. There were 6 differential expressed genes from all of these genes between the 2 G. bicolors with purple and green leaves, which were 4 significant down regulated genes including pelargonidin (c11692), cyanidin (c42112), anthocyanidin synthase (c38551), flavonoid 3 O glucosyltransferase (c9064) and 2 significant up regulated genes including dihydroflavonol (c35961), flavonoid 3 O glucosyltransferase (c20283). (2) The up or down expression trend of these 6 genes agreed very well with the testing by the RT PCR. However, the expression trend by transcriptome sequencing was more significantly different than that by qRT PCR. (3) According to the HPLC detection, the result showed that there were not 4 kinds of anthocyanidins such as delphinidin, peonidin, petunidin and malvidin in G. bicolor. However, the purple leaves had significantly higher content of cyanidin (62.21 mg/kg) than that in green leaves (6.86 mg/kg). At the same time, the content of pelargonidin was below 0.43 mg/kg in 2 G. bicolors. So this study inferred that the reason why the levels of the 2 kinds of anthocyanidins including cyanidin and pelargonidin were decreased significantly in green G. bicolor was the 4 anthocyanin synthesis regulator genes involved in positive regulatory genes like an anthocyanidin synthase and a flavonoid 3 O glucosyltransferase, negative regulatory genes like a dihydroflavonol and a flavonoid 3 O glucosyltransferase.
keywords:Gynura bicolor  transcriptome sequence  anthocyanin  anthocyanidins  regulator genes
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