Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur 176061, HP, India; Academy of Scientific and Innovative Research, New Delhi, India.
Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur 176061, HP, India; Academy of Scientific and Innovative Research, New Delhi, India.
Gene. 2014 Apr 15;539(2):250-7. doi: 10.1016/j.gene.2014.01.071. Epub 2014 Feb 8.
The transcript expression of a gene SrUGT85C2 has been documented for direct relation with steviol glycoside content in Stevia plant. Steviol glycoside and gibberellin biosynthetic routes are divergent branches of methyl erythritol-4 phosphate (MEP) pathway. So, SrUGT85C2 might be an influencing gibberellin content. Hence in the present study, transgenic Arabidopsis thaliana overexpressing SrUGT85C2 cDNA from Stevia rebaudiana was developed to check its effect on gibberellin accumulation and related plant growth parameters. The developed transgenics showed a noteworthy decrease of 78-83% in GA3 content. Moreover, the transgenics showed a gibberellin deficient phenotype comprising stunted hypocotyl length, reduced shoot growth and a significant fall in relative water content. Transgenics also showed 17-37 and 64-76% reduction in chlorophyll a and chlorophyll b contents, respectively. Reduction in photosynthetic pigments could be responsible for the noticed significant decrease in plant biomass. Like steviol glycoside and gibberellin biosynthesis, chlorophyll biosynthesis also occurs from the precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) of MEP pathway in the plastids. The observed downregulated expression of genes encoding MEP pathway enzymes geranyl geranyl diphosphate synthase (GGDPS), copalyl diphosphate synthase (CDPS), kaurenoic acid oxidase (KAO), chlorophyll synthetase and chlorophyll a oxygenase in transgenics overexpressing SrUGT85C2 might be responsible for the reduction in gibberellins as well as chlorophyll. This study has documented for the first time the regulatory role of SrUGT85C2 in the biosynthesis of steviol glycoside, gibberellins and chlorophyll.
基因 SrUGT85C2 的转录表达与甜叶菊植物中甜菊糖苷的含量有直接关系。甜菊糖苷和赤霉素的生物合成途径是甲基赤藓醇-4-磷酸(MEP)途径的两个不同分支。因此,SrUGT85C2 可能会影响赤霉素的含量。因此,本研究通过构建甜叶菊 SrUGT85C2 基因的转基因拟南芥,检测其对赤霉素积累和相关生长参数的影响,来研究 SrUGT85C2 的功能。结果表明,转基因植株中 GA3 的含量显著降低了 78%-83%。此外,转基因植株表现出赤霉素缺乏的表型,包括下胚轴伸长变短、地上部分生长减少以及相对含水量显著降低。转基因植株中叶绿素 a 和叶绿素 b 的含量也分别降低了 17%-37%和 64%-76%。光合色素的减少可能是导致植物生物量显著下降的原因。与甜菊糖苷和赤霉素的生物合成一样,叶绿素的生物合成也发生在质体中的 MEP 途径前体异戊烯二磷酸(IPP)和二甲基烯丙基二磷酸(DMAPP)上。在 SrUGT85C2 过表达的转基因植株中,编码 MEP 途径酶香叶基香叶基二磷酸合酶(GGDPS)、牻牛儿基牻牛儿基二磷酸合酶(CDPS)、贝壳杉烯酸氧化酶(KAO)、叶绿素合酶和叶绿素 a 加氧酶的基因表达下调,这可能是导致赤霉素和叶绿素减少的原因。本研究首次证明了 SrUGT85C2 在甜菊糖苷、赤霉素和叶绿素生物合成中的调控作用。