毛竹LhcaPe03基因在不同组织中的表达量
|
摘要
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1 149 bp cDNA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 1149 bp 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.
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 1149 bp 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.
引文
[1]訾兴中,李树春.安徽树木志[M].北京:中国林业出版社,2015.
[2]郁飞,唐崇钦,辛越勇,等.光系统I(PSI)的结构与功能研究进展[J].植物学通报,2001,18(3):266-275.
[3]The properties of the chorophyll a/b-binding proteins Lhca2 and Lhca3 using antisense inhibition[J].Plant Physiology,2001,127(1):150-158.
[4]FRANK K,ANDREAS S,CHRISTOS N,et al.Abundantly and rarely expressed Lhc protein genes exhibit distinct regulation patterns in plantsp[J].Plant Physiology,2006,140(3):793-804.
[5]张立国,张琚.实时定量PCR技术的介绍[J].生物技术,2003,13(2):39-40.
[6]彭书明,唐琳,叶杨,等.苦瓜BAG基因组织特异性表达研究[J].园艺学报2006,33(5):1007-1010.
[7]程占超,马艳军,侯丹,等.毛竹PheMADS15基因的克隆及功能分析[J].浙江农林大学学报,2017,34(3):421-426.
[8]王萍,丛敏,李忆梅,等.Taqman荧光定量PCR是核酸水平对管家基因GAPDH进行定量的好方法[J].首都医科大学学报,2006,27(2):210-213.
[9]唐文莉,高健彭,彭镇华.刚竹属3种重要散生竹光系统Ⅰ基因(Lhca1)的克隆、序列分析和蛋白结构预测[J].北京林业大学学报,2008,30(4):109-115.
[10]唐文莉,高健彭,彭镇华.毛竹(Phyllostachys edulis)光系统I基因LhcaPe02全长的克隆与序列分析[J].安徽农业大学学报,2008,35(2):153-15.
[11]黄培堂.分子克隆实验指南[M].科学出版社.2002:27-28.
[12]高志民,李雪平,彭镇华,等.竹子捕光叶绿素a/b结合蛋白基因全长的克隆和序列分析[J].林业科学.2007,43(3):34-38.
[13]马荣群,陈红运,黄文胜,等.实时定量PCR方法检测转基因产品[J].植物检疫,2002,16(1):61-64.
[2]郁飞,唐崇钦,辛越勇,等.光系统I(PSI)的结构与功能研究进展[J].植物学通报,2001,18(3):266-275.
[3]The properties of the chorophyll a/b-binding proteins Lhca2 and Lhca3 using antisense inhibition[J].Plant Physiology,2001,127(1):150-158.
[4]FRANK K,ANDREAS S,CHRISTOS N,et al.Abundantly and rarely expressed Lhc protein genes exhibit distinct regulation patterns in plantsp[J].Plant Physiology,2006,140(3):793-804.
[5]张立国,张琚.实时定量PCR技术的介绍[J].生物技术,2003,13(2):39-40.
[6]彭书明,唐琳,叶杨,等.苦瓜BAG基因组织特异性表达研究[J].园艺学报2006,33(5):1007-1010.
[7]程占超,马艳军,侯丹,等.毛竹PheMADS15基因的克隆及功能分析[J].浙江农林大学学报,2017,34(3):421-426.
[8]王萍,丛敏,李忆梅,等.Taqman荧光定量PCR是核酸水平对管家基因GAPDH进行定量的好方法[J].首都医科大学学报,2006,27(2):210-213.
[9]唐文莉,高健彭,彭镇华.刚竹属3种重要散生竹光系统Ⅰ基因(Lhca1)的克隆、序列分析和蛋白结构预测[J].北京林业大学学报,2008,30(4):109-115.
[10]唐文莉,高健彭,彭镇华.毛竹(Phyllostachys edulis)光系统I基因LhcaPe02全长的克隆与序列分析[J].安徽农业大学学报,2008,35(2):153-15.
[11]黄培堂.分子克隆实验指南[M].科学出版社.2002:27-28.
[12]高志民,李雪平,彭镇华,等.竹子捕光叶绿素a/b结合蛋白基因全长的克隆和序列分析[J].林业科学.2007,43(3):34-38.
[13]马荣群,陈红运,黄文胜,等.实时定量PCR方法检测转基因产品[J].植物检疫,2002,16(1):61-64.