《植物生理学报》 2021, 57(2): 293-300

拟南芥线粒体RNA加工蛋白(RPF)基因的鉴定及其功能和应用

赵重燕¹，暴会会¹，张丽梅^2,3，梁晓丽^2,4，杜康华¹，杨正安^1,*，杨飞^2,4,*

¹云南农业大学园林园艺学院, 昆明650201；²云南农业大学烟草学院, 昆明650201；³红云红河烟草(集团)有限责任公司昆明卷烟厂, 昆明650202；⁴云南农业大学云南生物资源保护与利用国家重点实验室, 昆明650201

通信作者：杨正安；E-mail: yangzhengan@ynau.edu.cn；杨飞；E-mail: feiyang1212@163.com

摘　要：

细胞质雄性不育(cytoplasmic male sterility, CMS)在植物杂交育种中广为应用, 其发生与线粒体中的CMS基因有着密切关系。但是由于线粒体基因敲除或下调表达技术手段十分有限, 各种作物中大量鉴定出的CMS基因都未能进行功能验证。拟南芥(Arabidopsis thaliana)线粒体RPF (RNA processing factor)蛋白属于PPR (pentatricopeptide repeat)蛋白家族中的P亚类, 由6~8个PPR重复基序组成。每个PPR基序含有35个氨基酸, 其第5位和第35位氨基酸对于识别线粒体靶向RNA的一个碱基起到关键作用, 由此建立了“PPR密码”规则。拟南芥RPF蛋白以序列识别的方式结合并加工线粒体RNA转录本的5′末端, 但目前尚不清楚该加工过程的具体细节及其生理功能。本文综述了RPF蛋白编码基因RPF1~RPF7的鉴定及其功能分析, 以及利用人工设计的RPF2蛋白使线粒体基因nad6下调表达的研究进展。植物线粒体RPF蛋白编码基因的鉴定及其功能和应用, 对于阐明线粒体mRNA 5′末端加工的分子机制, 以及利用反向遗传学手段研究线粒体基因功能, 具有重要的科学意义和应用前景。

关键词：拟南芥; 细胞质雄性不育; 线粒体RNA加工; PPR密码; RPF蛋白

收稿：2020-09-04   修定：2020-12-17

资助：国家自然科学基金(31960603和31760583)

Identifcation, function and application of mitochondrial RNA processing factor (RPF) genes in Arabidopsis thaliana

ZHAO Chongyan¹, BAO Huihui¹, ZHANG Limei^2,3, LIANG Xiaoli^2,4, DU Kanghua¹, YANG Zhengan^1,*, YANG Fei^2,4,*

¹College of Horticulture and Landscape, Yunnan Agricultural University, Kunming 650201, China; ²College of Tobacco Science, Yunnan Agricultural University, Kunming 650201, China; ³Kunming Cigarette Factory of Hongyun Honghe Tobacco (Group) Co., Ltd., Kunming 650202, China; ⁴State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming 650201, China

Corresponding author: YANG Zhengan; E-mail: yangzhengan@ynau.edu.cn; YANG Fei; E-mail: feiyang1212@163.com

Abstract:

Cytoplasmic male sterility (CMS) has been widely used in plant hybrid breeding and is related to CMS genes in mitochondria. Because limited methods are available to knockout or downregulate mitochondrial genes, it is difcult to verify the candidate CMS genes in most of the crops. Mitochondrial RNA processing factors (RPFs), which belong to the P subclass of pentatricopeptide repeat (PPR) proteins, are constituted by 8–16 PPR repeat motifs in Arabidopsis thaliana. The amino acids in the sequence position of 5 and 35 in each PPR repeat are key to recognize one base in the target mRNA, establishing the rule of “PPR code”. RPF proteins can bind and process the 5′ end of the target mitochondrial mRNA by sequence recognition manner in A. thaliana. However, little is known about the details of RPF function in the processing and their physiological effects. This article reviews the identifcation and functional analysis of seven RPF genes, which were named RPF1 to RPF7, and downregulation of mitochondrial nad6 by the redesigned RPF2 protein. Identifcation, functional analysis and application of the RPF genes have great signifcance in research and application prospect, to elucidate the molecular mechanism of mitochondrial mRNA 5′ end processing, and to study the function of mitochondrial genes via reverse genetics.

Key words: Arabidopsis thaliana; cytoplasmic male sterility; mitochondrial RNA processing; PPR code; RPF protein