《植物生理学报》 2015, 51(4): 501-508

拟南芥叶绿体分裂突变体cpd44中ARC6基因的鉴定与分析

张琴, 贾宁, 李慧, 高宏波*

北京林业大学生物科学与技术学院, 北京100083

通信作者：高宏波；E-mail: gaobjfu@yahoo.com；Tel: 010-62336496

摘　要：

叶绿体是光合作用的主要场所, 叶绿体的分裂对于维持其正常的功能有着非常重要的作用。本实验从EMS (ethyl methane sulfonate)诱变的拟南芥Columbia生态型(Col)种子中筛选到一个叶绿体分裂突变体cpd44 (chloroplast division 44), 其叶绿体的体积显著增大, 每个叶肉细胞中的叶绿体数目减少至1~5个。遗传分析显示该表型是单基因控制的隐性突变。运用图位克隆技术和测序分析发现ARC6 (accumulation and replication of chloroplasts 6)基因发生突变, 第1 160个碱基由C突变成T, 发生无义突变。半定量RT-PCR分析结果显示cpd44中ARC6的mRNA可能被降解。遗传互补实验进一步证明cpd44的叶绿体异常表型是由ARC6突变造成的。本工作为进一步研究叶绿体分裂机制提供了新材料。

关键词：拟南芥; 叶绿体分裂; ARC6; cpd44

收稿：2015-01-20   修定：2015-03-20

资助：高等学校学科创新引智计划(B13007)。

Identification and Analysis of ARC6 Gene in a Chloroplast Division Mutant cpd44 in Arabidopsis thaliana

ZHANG Qin, JIA Ning, LI Hui, GAO Hong-Bo*

College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China

Corresponding author: GAO Hong-Bo; E-mail: gaobjfu@yahoo.com; Tel: 010-62336496

Abstract:

Chloroplasts are the major place for photosynthesis. The division of chloroplast is important for the maintaining of its function. chloroplast division 44 (cpd44) is a chloroplast division mutant obtained in a mutant screening of ethyl methane sulfonate (EMS) mutagenized Arabidopsis thaliana in Col ecotype, in which the chloroplast volume was increased obviously and the chloroplast number was decreased to one to five per mesophyll cell. Genetic analysis indicated that the phenotype of cpd44 was controlled by a single recessive gene. Map-based cloning and sequencing analysis revealed a mutation in accumulation and replication of chloroplasts 6 (ARC6) gene, which was a change from C to T at the 1 160th base and resulted in a nonsense mutation. Semi-quantitative RT-PCR analysis suggested that the mRNA of ARC6 in cpd44 might be degraded. Genetic complementation experiment further confirmed that the chloroplast phenotype in cpd44 was caused by the mutation of ARC6. This work provides a new material for the further study of the division mechanism of chloroplast.

Key words: Arabidopsis thaliana; chloroplast division; ARC6; cpd44