建兰实时荧光定量PCR内参基因的筛选

GUO Li-ting; CHEN Juan; LI Yao; LI Jin-jin; TIAN Yang; AI Ye

doi:10.3969/j.issn.1009-7791.2024.05.005

亚热带植物科学 >

2024 , Vol. 53 >Issue 5: 425 - 432

DOI: https://doi.org/10.3969/j.issn.1009-7791.2024.05.005

研究论文

建兰实时荧光定量PCR内参基因的筛选

GUO Li-ting ,
CHEN Juan ,
LI Yao ,
LI Jin-jin ,
TIAN Yang ,
AI Ye

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(福建农林大学风景园林与艺术学院 / 兰科植物保护与利用国家林业与草原局重点实验室，福建福州 350002)

收稿日期: 2024-05-07

录用日期: 2024-07-29

网络出版日期: 2025-01-25

基金资助

福建农林大学科技创新专项基金项目(KFb22057XA)；国家重点研发计划课题(2019YFD1000401)

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Selection of Reference Genes for Quantitative Real-Time PCR in Cymbidium ensifolium

郭丽婷，陈娟，李瑶，李进金，田阳，艾叶

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(College of Landscape Architecture, Fujian Agriculture and Forestry University / Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization, Fuzhou 350002, Fujian China)

Received date: 2024-05-07

Accepted date: 2024-07-29

Online published: 2025-01-25

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摘要

为筛选出建兰Cymbidium ensifolium实时荧光定量PCR中稳定表达的内参基因，以建兰品种‘大凤素’的不同器官组织(花器官：萼片、捧瓣、唇瓣、合蕊柱；营养器官：根、假鳞茎、叶)为材料，利用实时荧光定量PCR技术分析5个候选内参基因(Actin、18SrRNA、rpoB、GAPDH、EF-1a)在‘大凤素’不同器官组织中的表达情况，利用geNorm、NormFinder、Bestkeeper软件对各候选内参基因的表达稳定性进行综合评价，并对筛选出的最佳内参基因进行验证。结果表明，候选内参基因在建兰‘大凤素’不同器官组织中表达稳定性存在差异，GAPDH和Actin分别为花器官和营养器官组织的最稳定内参基因。该研究可为建兰基因表达研究中内参基因的选择提供参考。

关键词： 建兰; 内参基因; 实时荧光定量PCR

本文引用格式

GUO Li-ting , CHEN Juan , LI Yao , LI Jin-jin , TIAN Yang , AI Ye . 建兰实时荧光定量PCR内参基因的筛选[J]. 亚热带植物科学, 2024 , 53(5) : 425 -432 . DOI: 10.3969/j.issn.1009-7791.2024.05.005

Abstract

In this study, different organ tissues (floral organs: sepal, petal, lip and gynostemium; vegetative organs: root, pseudobulb, and leaf) of Cymbidium ensifolium ‘Dafengsu’ were used as materials, to screen out the stably expressed reference genes for real-time quantitative PCR (qRT-PCR). The expression of five candidate internal reference genes (Actin, 18SrRNA, rpoB, GAPDH, EF-1a) in different organ tissues of ‘Dafengsu’ was analyzed by qRT-PCR. The expression stability of candidate reference genes was comprehensively evaluated by geNorm, NormFinder and Bestkeeper, and the selected reference genes were verified. The results indicated that the expression stability of candidate reference genes was varied, and GAPDH and Actin were the best stable reference genes in floral and vegetative organs in different parts of ‘Dafengsu’, respectively. This study can provide reference for the selection of reference genes in the gene expression research of C. ensifolium.

Key words： Cymbidium ensifolium; reference genes; qRT-PCR

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