多肉植物法师系品种qRT-PCR内参基因筛选  

Selection of qRT-PCR internal reference gene for succulent plants of Aeoniumseries

作　　者：赵荣 李素华[1] 韩浩章[1] 张丽华[1] 王芳[1] 蒋亚华[1] ZHAO Rong;LI Suhua;HAN Haozhang;ZHANG Lihua;WANG Fang;JIANG Yahua(Suqian University,Suqian,Jiangsu 223800,China)

机构地区：[1]宿迁学院,江苏宿迁223800

出　　处：《植物生理学报》2024年第4期714-724,共11页Plant Physiology Journal

基　　金：宿迁学院人才引进科研启动基金项目(校2022XRC036);宿迁学院重点建设学科项目(2021ZDJS04);宿迁学院创新团队项目(2021td05);宿迁市科技计划项目(K202207);宿迁市科技计划项目(M202001);苏北科技专项(SZ-SQ2021011)。

摘　　要：法师系多肉植物隶属于景天科莲花掌属(Aeonium),具有多个彩叶品种,观赏价值较高。然而,国内外还鲜有此类植物分子生物学方面的研究。为筛选出在不同法师系品种间稳定表达的内参基因,本研究以5个法师系品种叶片为材料,以法师系品种‘墨法师’和‘绿羊绒’转录组库为依据,筛选出了8个候选的qRT-PCR (quantitative RT-PCR)内参基因,并通过qRT-PCR分析了这些内参在不同品种间的表达丰度,经Genorm、normFinder、BestKeeper分析和综合分析,最终筛选出了法师系不同品种间的最适内参基因GAPDH。以该基因为内参基因,对‘墨法师’和‘绿羊绒’转录组数据库中差异表达的花青素合成酶相关基因进行表达水平验证,结果显示,除了AaCHS外,其他6个基因在上述2个品种中确实存在差异表达,进一步表明GAPDH适合作为法师系不同品种间基因表达分析的内参。本研究可为在法师系不同品种间进行基因表达分析奠定坚实的基础。The Aeonium series succulent plants belong to the sedum family,with multiple colored leaf vari-eties and high ornamental value.However,there are few studies on the molecular biology of such plants at home and abroad.In order to screen out the internal reference genes that are stably expressed among dif-ferent cultivated varieties of Aeonium series succulent plants,the leaves of five cultivated varieties of the Aeonium series were used as materials,and eight candidate qRT-PCR(quantitative RT-PCR)reference genes were selected based on the transcriptome libraries of the cultivated varieties Aeonium arboreum‘Zwartkop'and Aeonium spp..The expression abundance of these internal reference genes among differ-ent cultivated varieties was analyzed by qRT-PCR.After Genorm,normFinder,BestKeeper analysis and comprehensive analysis,the most suitable internal reference gene GAPDH among different varieties of the Aeonium series succulent plants was finally screened.Using this gene as an internal reference gene,the differentially expressed genes of anthocyanin synthesis in the transcriptome libraries of Aeonium arboreum‘Zwartkop'and Aeonium spp.were verified.The results showed that except for AaCHS,the other six genes are indeed differentially expressed in the above two varieties,indicating that GAPDH is a suitable internal reference gene for expression analysis among different cultivated varieties of Aeonium series succulent plants.This study can lay a solid foundation for gene expression analysis among different varieties of Aeonium series succulent plants.

关 键 词：法师系多肉植物 内参基因 荧光定量PCR 

分 类 号：S682.33[农业科学—观赏园艺]