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水稻胆碱单加氧酶(OsCMO)蛋白在增强转基因烟草中海藻糖胺生物合成中发挥作用,但在水稻(Oryza sativa L. ssp. japonica)中并不积累。

Rice choline monooxygenase (OsCMO) protein functions in enhancing glycine betaine biosynthesis in transgenic tobacco but does not accumulate in rice (Oryza sativa L. ssp. japonica).

机构信息

Ministry of Education Key Laboratory for Bio-resource and Eco-environment, College of Life Science and State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu, 610064, China.

出版信息

Plant Cell Rep. 2012 Sep;31(9):1625-35. doi: 10.1007/s00299-012-1276-2. Epub 2012 May 9.

Abstract

UNLABELLED

Glycine betaine (GB) is a compatible quaternary amine that enables plants to tolerate abiotic stresses, including salt, drought and cold. In plants, GB is synthesized through two-step of successive oxidations from choline, catalyzed by choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH), respectively. Rice is considered as a typical non-GB accumulating species, although the entire genome sequencing revealed rice contains orthologs of both CMO and BADH. Several studies unraveled that rice has a functional BADH gene, but whether rice CMO gene (OsCMO) is functional or a pseudogene remains to be elucidated. In the present study, we report the functional characterization of rice CMO gene. The OsCMO gene was isolated from rice cv. Nipponbare (Oryza sativa L. ssp. japonica) using RT-PCR. Northern blot demonstrated the transcription of OsCMO is enhanced by salt stress. Transgenic tobacco plants overexpressing OsCMO results in increased GB content and elevated tolerance to salt stress. Immunoblotting analysis demonstrates that a functional OsCMO protein with correct size was present in transgenic tobacco but rarely accumulated in wild-type rice plants. Surprisingly, a large amount of truncated proteins derived from OsCMO was induced in the rice seedlings in response to salt stresses. This suggests that it is the lack of a functional OsCMO protein that presumably results in non-GB accumulation in the tested rice plant.

KEY MESSAGE

Expression and transgenic studies demonstrate OsCMO is transcriptionally induced in response to salt stress and functions in increasing glycinebetaine accumulation and enhancing tolerance to salt stress. Immunoblotting analysis suggests that no accumulation of glycinebetaine in the Japonica rice plant presumably results from lack of a functional OsCMO protein.

摘要

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甘氨酸甜菜碱(GB)是一种相容的季铵盐,可使植物耐受非生物胁迫,包括盐、干旱和寒冷。在植物中,GB 通过两步连续氧化由胆碱合成,分别由胆碱单加氧酶(CMO)和甜菜碱醛脱氢酶(BADH)催化。水稻被认为是一种典型的非 GB 积累物种,尽管全基因组测序表明水稻含有 CMO 和 BADH 的直系同源物。几项研究表明,水稻具有功能性 BADH 基因,但水稻 CMO 基因(OsCMO)是否具有功能还是假基因仍有待阐明。在本研究中,我们报告了水稻 CMO 基因的功能特征。使用 RT-PCR 从水稻品种 Nipponbare(Oryza sativa L. ssp. japonica)中分离出 OsCMO 基因。Northern blot 表明 OsCMO 的转录受盐胁迫增强。过表达 OsCMO 的转基因烟草植物增加了 GB 含量并提高了对盐胁迫的耐受性。免疫印迹分析表明,功能性 OsCMO 蛋白存在于转基因烟草中,但在野生型水稻植物中很少积累。令人惊讶的是,大量源自 OsCMO 的截断蛋白在盐胁迫下诱导水稻幼苗中表达。这表明,缺乏功能性 OsCMO 蛋白可能导致所测试的水稻植物中不积累 GB。

关键信息

表达和转基因研究表明,OsCMO 受盐胁迫转录诱导,可增加甘氨酸甜菜碱的积累并增强对盐胁迫的耐受性。免疫印迹分析表明,日本水稻植物中甘氨酸甜菜碱的积累量低可能是由于缺乏功能性 OsCMO 蛋白所致。

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