茶叶科学Issue(1)：92-101,10.
不同氮处理茶树实时定量 PCR 内参基因筛选和验证
Screening and Validation of Reference Genes for Quantitative Real-time PCR Analysis in Tea Plant (Camellia sinensis) under Different Nitrogen Nutrition
摘要
Abstract
The objective of this study was to select the most reliable reference genes for qRT-PCR analysis of target tea plant genes under varying nitrogen source and availability. We chose 6 housekeeping genes which included five commonly used and one new candidates to systematically assess their expression levels at three different tissues (young leaves, mature leaves and roots) under different nitrogen regimes by qRT-PCR. GeNorm and NormFinder software were used to analyze and evaluate the data for reference genes. The results indicated that GAPDH, β-actin and RPL13 are the best reference genes for normalizing target gene expression in tea plant under different nitrogen nutrition, whereas a-tubulin and RuBP are not suitable in many experimental conditions and the best combination (GAPDH+β-actin) was recommended. Meanwhile, the expression levels of CsNRT1.2 and CsAMT1.1 in young leaves of tea plants were analyzed. The results showed that the variation tendency of CsNRT1.2 and CsAMT1.1 are exactly consistent when using GAPDH and GAPDH+β-actin as reference genes. However, the expression levels of these genes are showed significant differences when a-tubulin was used as a reference gene. Thus, validation of suitable reference genes for specific condition can guarantee the accurate quantification of the target genes in qRT-PCR analysis.关键词
茶树/内参基因/氮处理/qRT-PCRKey words
tea plant (Camellia sinensis)/reference gene/nitrogen treatment/qRT-PCR分类
农业科技引用本文复制引用
刘圆,王丽鸳,韦康,成浩,张芬,吴立赟,胡娟..不同氮处理茶树实时定量 PCR 内参基因筛选和验证[J].茶叶科学,2016,(1):92-101,10.基金项目
浙江省茶树农业新品种选育重大科技专项（2012C12905）、国家茶叶产业技术体系（nycytx-23）。 （）
