黑龙江省青年科学基金(QC2016037) ;国家国际科技合作专项(2015DFR50260)
大麻THCA合成酶基因CsTHCA在调控大麻THC含量方面起着重要的作用。通过RT-PCR方法从大麻中克隆了CsTHCA,该基因开放阅读框全长1638 bp,共编码545个氨基酸。通过pSKint中间载体,构建CsTHCA RNA干扰植物转化载体并转化大麻茎尖,获得转基因植株,通过分析大麻中THCA合成酶基因的表达并结合薄层色谱法和SPE-HPLC法发现,转基因植株中基因表达量降低且THC含量降低,这表明CsTHCA通过某种机理正调控THC的含量。
CsTHCA plays important role in regulating THC contents. In this study, we isolated the complete coding sequence of CsTHCA from Cannabis Sativa L., with an open reading frame of 1638 base pair. The deduced CsTHCA protein contained 545 amino acids. The CsTHCA RNA interference construct was generated through pSKint intermediate vector, which was transformed into the shoot tips of hemp. By analyzing the transcriptional level of CsTHCA by semi-quantitative RT-PCR and the THC content by thin-layer chromatography and SPE-HPLC, the reduction of CsTHCA expression and THC content was observed in transgenic lines relative to wild type. Thus, our results supported a positive role of CsTHCA in modulating THC content in hemp.
姜颖,孙宇峰,王晓楠,等.大麻THCA合成酶基因(CsTHCA)RNA干扰载体的构建及遗传转化[J].植物遗传资源学报,2019,20(1):207-214.
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