《植物生理学报》 2021, 57(1): 225-234

屏边空竹实时荧光定量PCR内参基因的筛选

李露双，陈凌娜，夏体泽，杨汉奇^*

中国林业科学研究院资源昆虫研究所, 昆明650224

通信作者：杨汉奇；E-mail: yanghanqikm@aliyun.com

摘　要：

屏边空竹(Cephalostachyum pingbianense)是目前已知唯一在自然条件下能够四季产笋的竹(sf. Bambusoideae)种, 对于研究竹笋芽萌发机理的多样性有重要意义。为筛选出在屏边空竹中稳定表达的内参基因, 本研究利用实时荧光定量PCR技术分析6个常见内参基因(GAPDH、PP2A、EF1α、TIP41、RPL3、ACT1)在屏边空竹不同组织器官(根、秆、叶)、不同发育时期(休眠期、萌动期、发育期、成熟期)的笋芽以及处于快速生长期的竹笋中的表达情况, 并采用geNorm、NormFinder、BestKeeper软件评价各候选内参基因的稳定性。结果表明: 在不同的组织器官中, 表达最稳定的基因为EF1α和ACT1; 在笋芽发育成笋的过程中, 表达最稳定的基因为ACT1和RPL3。ACT1和RPL3在所有样本中均表现出较高的稳定性, 且ACT1的表达水平较高, 可用于校正较高表达水平目的基因; RPL3的表达水平较低, 适用于校正较低表达水平目的基因。

关键词：屏边空竹; 内参基因; 实时荧光定量PCR

收稿：2020-09-23   修定：2020-11-19

资助：国家自然科学基金(31870574)

Selection of reference genes for quantitative real-time PCR analysis in Cephalostachyum pingbianense

LI Lushuang, CHEN Lingna, XIA Tize, YANG Hanqi^*

Research Institute of Resources Insects, Chinese Academy of Forestry, Kunming 650224, China

Corresponding author: YANG Hanqi; E-mail: yanghanqikm@aliyun.com

Abstract:

Cephalostachyum pingbianense is the only known bamboo (sf. Bambusoideae) species that can produce bamboo shoots in four seasons under natural conditions, which is of great significance for the study of bamboo shoots germination mechanism. In order to choose suitable reference genes in C. pingbianense, quantitative real-time PCR was used to analyze the expression stability of six candidate genes (GAPDH, PP2A, EF1α, TIP41, RPL3 and ACT1) in different tissues (root, stem and leaf), shoot bud at different developmental stages (dormant stage, germinative stage, developmental stage and mature stage), and bamboo shoot at fast growth stage. geNorm, NormFinder and BestKeeper were used to evaluate the stability of candidate reference genes. The results show that EF1α and ACT1 were the most stable genes in different tissues, and ACT1 and RPL3 were most stable in the developmental stages from shoot bud to shoot. ACT1 and RPL3 showed high stability in all samples. The expression abundance of ACT1 was high, which could be used for the correction of target genes with high expression level. And RPL3 showed low expression abundance in all samples, which is suitable for the correction of target genes with low expression level.

Key words: Cephalostachyum pingbianense; reference gene; quantitative real-time PCR