《植物生理学报》 2010, 46(10): 1061-1066

可用于实时荧光定量PCR 标准化的白桦内参基因

尹静¹, 任春林¹, 詹亚光^1,*, 滕文华², 陈秀福¹, 王玉成³, 孙红冉¹

东北林业大学1 生命学院; 2 帽儿山实验林场, 3 林学院, 哈尔滨150040

通信作者：詹亚光；E-mail: yaguangzhan@126.com；Tel: 0451-82191752

摘　要：

为了快速、准确的对白桦多个样品进行基因表达分析, 本研究应用荧光定量PCR 技术, 对白桦持家基因微管蛋白基 因(Tu)和泛素基因(UbQ)在不同处理的细胞及不同树龄或不同生长季节白桦植株各部位表达的稳定性进行检测。结果表明, 对于茉莉酸甲酯(MeJA)、水杨酸(SA)处理的白桦细胞和不同生长季节白桦植株的基因定量表达分析, 可同时使用Tu和UbQ 平衡化 qRT-PCR 数据; 而针对不同树龄白桦茎皮中基因表达研究时, 单独使用 UbQ 基因作为内参对照即可获得精确的表 达数据。

关键词：白桦; 实时定量PCR; 持家基因

收稿：2010-05-13   修定：2010-07-30

资助：黑龙江省自然基金重点项目(ZD200918)和中央高校基本科研业务费专项资金(DL09BA14)。

Selection of Internal Control Genes for Real-Time RT-PCR Normalization in White Birch (Betula platyphylla Suk.)

YIN Jing¹, REN Chun-Lin¹, ZHAN Ya-Guang^1,*, TENG Wen-Hua², CHEN Xiu-Fu¹, WANG Yu-Cheng³, SUN Hong-Ran¹

¹College of Life Sciences, ²Maoershan Experimental Forestry Center, ³College of Forestry, Northeast Forestry University, Harbin 150040, China

Corresponding author: ZHAN Ya-Guang; E-mail: yaguangzhan@126.com; Tel: 0451-82191752

Abstract:

In order to facilitate rapid and accurate gene expression analysis of multiple samples from birch, the expression stability of housekeeping genes such as genes coding micro-tube protein (Tu) and ubiquitin (UbQ) were assessed by qRT-PCR using a set of cDNAs from different birch samples including cells after different treatments and different parts of birch plants in different ages or different growing seasons. The results suggested that the Tu and UbQ genes could provided superior transcript normalization in birch cells treated with different elicitors (MeJA and SA), and birch plants in different growing seasons, while UbQ gene was the recommended reference gene for gene expression studies using cDNAs from white birch bark in defferent years.

Key words: birch; real-time RT-PCR; housekeeping gene