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OsPPR6,一个参与编辑和拼接叶绿体 RNA 的五肽重复蛋白,是水稻叶绿体生物发生所必需的。

OsPPR6, a pentatricopeptide repeat protein involved in editing and splicing chloroplast RNA, is required for chloroplast biogenesis in rice.

机构信息

State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China.

Jiangsu Collaborative Innovation Center for Modern Crop Production, Nanjing, China.

出版信息

Plant Mol Biol. 2017 Nov;95(4-5):345-357. doi: 10.1007/s11103-017-0654-0. Epub 2017 Aug 30.

Abstract

OsPPR6, a pentatricopeptide repeat protein involved in editing and splicing chloroplast RNA, is required for chloroplast biogenesis in rice. The chloroplast has its own genetic material and genetic system, but it is also regulated by nuclear-encoded genes. However, little is known about nuclear-plastid regulatory mechanisms underlying early chloroplast biogenesis in rice. In this study, we isolated and characterized a mutant, osppr6, that showed early chloroplast developmental defects leading to albino leaves and seedling death. We found that the osppr6 mutant failed to form thylakoid membranes. Using map-based cloning and complementation tests, we determined that OsPPR6 encoded a new Pentatricopeptide Repeat (PPR) protein localized in plastids. In the osppr6 mutants, mRNA levels of plastidic genes transcribed by the plastid-encoded RNA polymerase decreased, while those of genes transcribed by the nuclear-encoded RNA polymerase increased. Western blot analyses validated these expression results. We further investigated plastidic RNA editing and splicing in the osppr6 mutants and found that the ndhB transcript was mis-edited and the ycf3 transcript was mis-spliced. Therefore, we demonstrate that OsPPR6, a PPR protein, regulates early chloroplast biogenesis and participates in editing of ndhB and splicing of ycf3 transcripts in rice.

摘要

OsPPR6,一个参与叶绿体 RNA 编辑和剪接的五肽重复蛋白,是水稻叶绿体生物发生所必需的。叶绿体有自己的遗传物质和遗传系统,但它也受核编码基因的调控。然而,关于核质调控机制在水稻早期叶绿体生物发生中的作用知之甚少。在本研究中,我们分离并鉴定了一个突变体 osppr6,该突变体表现出早期叶绿体发育缺陷,导致白化叶片和幼苗死亡。我们发现 osppr6 突变体无法形成类囊体膜。通过基于图谱的克隆和互补测试,我们确定 OsPPR6 编码一种新的五肽重复(PPR)蛋白,定位于质体中。在 osppr6 突变体中,由质体编码 RNA 聚合酶转录的质体基因的 mRNA 水平降低,而由核编码 RNA 聚合酶转录的基因的 mRNA 水平增加。Western blot 分析验证了这些表达结果。我们进一步研究了 osppr6 突变体中的质体 RNA 编辑和剪接,发现 ndhB 转录本发生了错误编辑,ycf3 转录本发生了错误剪接。因此,我们证明了 OsPPR6 是一种 PPR 蛋白,它调节水稻早期叶绿体生物发生,并参与 ndhB 的编辑和 ycf3 转录本的剪接。

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