茶树两个MYB转录因子基因的克隆及功能验证

doi:10.13305/j.cnki.jts.2014.01.005

摘要/Abstract

摘要： 第4亚组R2R3-MYB可能参与木质素合成的调控,从而影响植物的生长发育。本文利用RACE技术,克隆了两个茶树第4亚组MYB转录因子（CsMYB4-5和CsMYB4-6）。生物信息学分析发现CsMYB4-5氨基酸序列与金鱼草中的AmMYB330一致性为48.45%,与拟南芥中的AtMYB3一致性为44.79%;CsMYB4-6氨基酸序列与金鱼草中的AmMYB308一致性为69.80%,与拟南芥中的AtMYB4一致性为62.41%。实时荧光定量PCR分析表明两个基因均在根中高表达而在茎中低表达。原核表达分析表明,CsMYB4-5和CsMYB4-6的分子量分别为32 kD和27 kD左右。烟草转化实验表明,与野生型烟草相比较,转CsMYB4-6基因的烟草叶片的叶脉紧缩而脉间凹凸不平,老叶有白色斑点,而转CsMYB4-5基因的烟草老叶发黄。

关键词: 茶树, MYB转录因子, 表达分析, 原核表达分析, 烟草转化分析

Abstract: R2R3-MYB in Sg4 subgroup probably participated in the regulation of lignin biosynthesis, which possibly play an important physiological role in plant growth and development. Two MYB transcription factors CsMYB4-5 and CsMYB4-6 in Sg4 subgroup were cloned by RACE technology. Investigation showed that the amino acid sequence of CsMYB4-5 showed 48.45% identity to AmMYB330 from Antirrhinum majus and 44.79% to AtMYB3 from Arabidopsis thaliana, while CsMYB4-6 showed 69.80% identity to AmMYB308 from Antirrhinum majus and 62.41% to AtMYB4 from Arabidopsis thaliana by bioinformatics analysis. The result of real-time fluorescent quantitative PCR showed that these two genes had all high expression levels in root while low expression levels in stem. The analysis of prokaryotic expression revealed that the recombinant CsMYB4-5 and CsMYB4-6 were expressed with a molecular weight of about 32 kD and 27 kD respectively. Compared with wild tobacco, transgenic tobacco with CsMYB4-6 gene appeared following symptoms: the veins of leaf tighten, lamina surface between veins with sags and crests, with white freckles in old leaf, but in transgenic tobacco with CsMYB4-5 gene the elder leaves turned yellow.

Key words: Camellia sinensis (L.), MYB transcription factor, expression analysis, prokaryotic expression analysis, transgenic tobacco analysis

中图分类号:

S571.1
Q52

贡年娣, 郭丽丽, 王弘雪, 赵磊, 王婕, 王文钊, 刘亚军, 王云生, 高丽萍, 夏涛. 茶树两个MYB转录因子基因的克隆及功能验证[J]. 茶叶科学, 2014, 34(1): 36-44. doi: 10.13305/j.cnki.jts.2014.01.005.

GONG Niandi, GUO Lili, WANG Hongxue, ZHAO Lei, WANG Jie, WANG Wenzhao, LIU Yajun, WANG Yunsheng, GAO Liping, XIA Tao. Cloning and Functional Verification of Two MYB Transcription Factors in Tea Plant [Camellia sinensis (L.) ][J]. Journal of Tea Science, 2014, 34(1): 36-44. doi: 10.13305/j.cnki.jts.2014.01.005.

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