花椰菜花球中花青素合成相关基因RT-qPCR内参基因的筛选
Screening of Reference Genes for RT-qPCR Analysis of Anthocyanin Biosynthesis Related Genes in Cauliflower Curd
张冠 1杨迎霞 2陆国清 2陈锐 2王倩 2王梦梦 1李荣 3姚星伟4
作者信息
- 1. 天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津,300134;天津市农业科学院生物技术研究所,天津,300381
- 2. 天津市农业科学院生物技术研究所,天津,300381
- 3. 天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津,300134
- 4. 天津市农业科学院蔬菜研究所,天津,300381
- 折叠
摘要
花椰菜收获过程中花球变紫的现象与花青素含量密切相关,目前针对花椰菜花球里花青素合成途径中相关基因表达分析的内参基因尚未报道.本研究以花椰菜始终不变紫色的白色花球组织和变紫花球中的未变紫、浅紫和深紫花球组织为研究材料,采用RT-qPCR技术测定了 6个候选内参基因在不同颜色花椰菜花球组织中的表达水平,用ΔCt法以及GeNorm、NormFinder、BestKeeper和RefFinder软件分析候选内参基因的稳定性.结果表明:花椰菜不同颜色花球组织中,6个候选内参基因的稳定性综合排名为SKIP16＞A CT＞PP2A＞TUB＞UBQ＞EF1α.SKIP16基因综合稳定性排名第一,其次是A CT基因,均适合作为花椰菜花球组织中花青素合成途径相关基因研究的理想内参基因;geNorm软件通过计算配对变异值分析最适内参基因组合的数量为2个,综合选用SKIP16和ACT基因.本研究结果可作为分析花椰菜花球花青素合成途径中相关基因准确测定的参考依据.
Abstract
When cauliflower is harvested,the curd typically becomes purple,which is strongly related to the an-thocyanin content.To date,no reference genes for evaluating the expression of related genes in the anthocyanin production pathway in cauliflower have been discovered.White curd tissue in the none-turning purple curd,white curd,light purple curd and dark purple tissue in turning purple curd were employed as research materials in this study.The expression of 6 potential reference genes in various colored cauliflower curd was determined using RT-qPCR.The ΔCt method,GeNorm,NormFinder,Bestkeeper,and RefFinder software were used to assess the stability of potential reference genes.The stability of six putative reference genes in cauliflower was determined to be SKIP16＞ACT＞PP2A＞TUB＞UBQ＞EF1α.SKIP16 ranks in first in terms of overall stability,followed by ACT.They are suitable reference gene candidate for researching genes involved in the biosynthesis of anthocyanins in cauliflower.By calculating the paired variation value with GeNorm software,the best number of candidate refer-ence gene pairs was 2,and the SKIP16 and A CT genes were fully picked.The findings of this study can be utilized as a guide for accurately determining associated genes in the cauliflower anthocyanin synthesis pathway.
关键词
花椰菜/花青素/内参基因/RT-qPCRKey words
Cauliflower/Anthocyanin/Reference genes/RT-qPCR引用本文复制引用
基金项目
天津市科技计划(19ZXZYSN0050)
天津市科技计划(20YFZCSN00400)
现代农业产业技术体系建设项目(CARS-23-A07)
天津市蔬菜产业技术体系建设项目(ITTVRS2017004)
青年科研人员创新研究与实验项目(2021023)
出版年
2024
NETL
NSTL
万方数据