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大花铁线莲单重被花的转录组差异分析

花匠小妙招
2024-11-26 10:50

摘要： 以即可开单被花又可开重被花的大花铁线莲重被品种"薇安"为研究对象，对"薇安"同一时期同一植株上的3种不同花瓣类型（单被、半重被、重被）采用高通量测序技术进行拼接及功能注释，筛选不同花被类型下表达量高度差异的关键基因后进行实时荧光定量PCR验证。结果显示：转录组测序共产生13.8GB原始数据，3个转录本文库两两比较共获得3 075条差异表达基因（DEG），其中单被花与半重被样本对比（A vs B）包括649条上调DEG，605条下调DEG；半重被花与重被花对比（B vs C）包括上调DEG 1 046条和下调DEG 721条；单被花与重被花对比（A vs C）有上调DEG 1 129条和下调DEG 859条。3个不同花被下共存的差异表达基因有134条。根据基因功能注释从总DEG中筛选出26条可能与重瓣化性状相关的基因进行聚类分析，并随机挑选10个目的基因进行荧光定量PCR验证。PCR结果显示这些基因的表达量在铁线莲"薇安"同一时期同一植株的3种花被类型中均存在显著差异。最终筛选出与铁线莲重瓣化相关的关键基因有MADS-BOX类基因PMADS1、AP3、FRUITFULL、FLC；生长素反应蛋白IAA9、生长素输入载体、脱落酸8'羟化酶、吲哚乙酸诱导蛋白ARG7等。本研究为探究铁线莲重瓣花分子机制提供了基础数据和理论依据。

关键词: 大花铁线莲, 单重被花, 高通量测序, 荧光定量PCR

Abstract: With the typical Clematis patens ‘Vyvyan Pennell’ which can open either single petal or double petals as the research object, we chose high throughput sequencing to splice and annotate the sequencing data of three different petal types(single petal, semi-double petal and double petal) on the same plant at the same period of C.patens ‘Vyvyan Pennell’, then selected differentially expressed genes to real-time fluorescence quantitative PCR. There were 13.8 GB raw data. A total of 3 075 differentially expressed genes(DEG) were obtained from the three transcriptome libraries by paired comparison, including 649 upregulated DEG and 605 upregulated DEG in the comparison between single and semi-double petals samples(A vs B). The comparison of semi-double flowers with double flowers(B vs C) included 1 046 up-regulated DEG and 721 down-regulated DEG. There were 1 129 upregulation DEG and 859 upregulation DEG in the comparison between single and double flowering(A vs C). There were 134 differentially expressed genes coexisting under three different perianth. By gene functional annotation, 26 genes that might be related to the double petal trait were screened out from the total DEG for cluster analysis, and 10 target genes were randomly selected for fluorescence quantitative PCR verification. PCR results showed that the expression levels of these genes were significantly different in the three floral envelope types of the same plant at the same time of C.patens ‘Vyvyan Pennell’. Finally, the key genes associated with double tepals Clematis conversion were screened out, including MADS-BOX genes PMADS1, AP3, FRUITFULL and FLC. Auxin reactive protein IAA9, auxin input vector, abscisic acid 8' hydroxylase, indoleacetic acid-induced protein ARG7, etc. This study provides basic data and theoretical basis for exploring the molecular mechanism of Clematis double flower.

Key words: Clematis patens, single and double tepals, high throughput sequencing, fluorescence quantitative PCR

中图分类号:

S68