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。
R2R3-MYB in Sg4 subgroup probably participated in the regulation of liguln 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 flom Antirrhinum majus and 44.79% to AtMYB3 flom ~trabidopsis thaliana, while CsMYB4-6 showed 69.80% identity to AmMYB308 flom 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.
出处
《茶叶科学》
CAS
CSCD
北大核心
2014年第1期36-36,37-44,共9页
Journal of Tea Science
基金
国家自然科学基金(30972401
31170647
31170282)
安徽省自然科学基金(11040606M73)
安徽省高校自然科学基金(KJ2012A110)
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
Camellia sinensis （L.）, MYB transcription gactor, expression analysis, prokaryotic expression analysis,transgenic tobacco analysis
