毛竹LhcaPe03基因在不同组织中的表达量

Different Tissue-specific Expressions of Gene LhcaPe03 in Phyllostachys edulis

摘取毛竹苗幼嫩新鲜叶片,提取RNA,反转录进行RT-PCR,先克隆捕光叶绿素a/b结合蛋白基因Lhca3片段,采用RACE扩增全长,获得一条1149bpcDNA的基因,检测后定名为LhcaPe03。从毛竹未出土的笋芽、叶(顶端第一片新叶、老叶)、茎(茎尖、茎秆)中提取RNA备用,根据已测的毛竹LhcaPe03基因序列设计引物,采用实时荧光定量PCR技术,检测毛竹LhcaPe03基因在不同部位的相对表达量。结果显示:LhcaPe03基因在毛竹不同部位的表达差异明显,茎尖和新叶中表达水平较高,在老叶和茎秆中表达量相当低,在笋中几乎没有表达。这说明毛竹新叶的光合能力比老叶强;茎尖和茎秆呈绿色,具有光合能力,茎尖光合能力较强;未出土的笋芽几乎不参加光合。

RNA was extracted from bamboo shoots tender leaves, and underwent reverse transcription RT-PCR. A segment of chlorophyll a-b binding protein LhcaPe03 gene was first cloned, and then an 1149bp cDNA was obtained through RACE, which was named LhcaPe03. RNA's in earth-covered bamboo shoots, leaves and stalks were extracted for backups. Primers were designed according to determined sequence of LhcaPe03 gene, and the expressions of LhcaPe03 gene in different tissues of Phyllostachys pubescens was determined by real-time quantitative fluorescence PCR. The results showed that the expression of LhcaPe03 gene in different tissues was significantly different, with expression of LhcaPe03 gene in shoot tip and new leaf being higher than that in old leaf, stem and stalk, and with almost no expression in shoot. These indicated that new leaves of Phyllostachys pubescens had stronger photosynthetic ability than old leaves did. The green shoot tip and stalk had photosynthetic ability, with shoot tip's photosynthetic ability being stronger;the earth-covered shoots almost did not participate in photosynthesis.