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一个甘薯香叶基香叶基焦磷酸合酶基因IbGGPS增加拟南芥类胡萝卜素含量并增强其渗透胁迫耐受性。

A Sweetpotato Geranylgeranyl Pyrophosphate Synthase Gene, IbGGPS, Increases Carotenoid Content and Enhances Osmotic Stress Tolerance in Arabidopsis thaliana.

作者信息

Chen Wei, He Shaozhen, Liu Degao, Patil Gunvant B, Zhai Hong, Wang Feibing, Stephenson Troy J, Wang Yannan, Wang Bing, Valliyodan Babu, Nguyen Henry T, Liu Qingchang

机构信息

Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education, China Agricultural University, Beijing, China.

Division of Plant Sciences and National Center for Soybean Biotechnology, University of Missouri, Columbia, Missouri, United States of America.

出版信息

PLoS One. 2015 Sep 16;10(9):e0137623. doi: 10.1371/journal.pone.0137623. eCollection 2015.

Abstract

Sweetpotato highly produces carotenoids in storage roots. In this study, a cDNA encoding geranylgeranyl phyrophosphate synthase (GGPS), named IbGGPS, was isolated from sweetpotato storage roots. Green fluorescent protein (GFP) was fused to the C-terminus of IbGGPS to obtain an IbGGPS-GFP fusion protein that was transiently expressed in both epidermal cells of onion and leaves of tobacco. Confocal microscopic analysis determined that the IbGGPS-GFP protein was localized to specific areas of the plasma membrane of onion and chloroplasts in tobacco leaves. The coding region of IbGGPS was cloned into a binary vector under the control of 35S promoter and then transformed into Arabidopsis thaliana to obtain transgenic plants. High performance liquid chromatography (HPLC) analysis showed a significant increase of total carotenoids in transgenic plants. The seeds of transgenic and wild-type plants were germinated on an agar medium supplemented with polyethylene glycol (PEG). Transgenic seedlings grew significantly longer roots than wild-type ones did. Further enzymatic analysis showed an increased activity of superoxide dismutase (SOD) in transgenic seedlings. In addition, the level of malondialdehyde (MDA) was reduced in transgenics. qRT-PCR analysis showed altered expressions of several genes involved in the carotenoid biosynthesis in transgenic plants. These data results indicate that IbGGPS is involved in the biosynthesis of carotenoids in sweetpotato storage roots and likely associated with tolerance to osmotic stress.

摘要

甘薯在块根中大量合成类胡萝卜素。在本研究中,从甘薯块根中分离出一个编码牻牛儿基牻牛儿基焦磷酸合酶(GGPS)的cDNA,命名为IbGGPS。绿色荧光蛋白(GFP)与IbGGPS的C末端融合,获得IbGGPS - GFP融合蛋白,该蛋白在洋葱表皮细胞和烟草叶片中瞬时表达。共聚焦显微镜分析确定IbGGPS - GFP蛋白定位于洋葱质膜的特定区域和烟草叶片的叶绿体中。将IbGGPS的编码区克隆到35S启动子控制下的二元载体中,然后转化到拟南芥中获得转基因植株。高效液相色谱(HPLC)分析表明转基因植株中总类胡萝卜素显著增加。转基因和野生型植株的种子在添加聚乙二醇(PEG)的琼脂培养基上萌发。转基因幼苗的根比野生型幼苗长得明显更长。进一步的酶活性分析表明转基因幼苗中超氧化物歧化酶(SOD)的活性增加。此外,转基因植株中丙二醛(MDA)的水平降低。qRT - PCR分析表明转基因植株中几个参与类胡萝卜素生物合成的基因表达发生了改变。这些数据结果表明IbGGPS参与甘薯块根中类胡萝卜素的生物合成,并且可能与渗透胁迫耐受性有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09b0/4574098/bd5d98196ec9/pone.0137623.g001.jpg

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