花生FAD2基因RNAi载体转化及转基因籽粒脂肪酸分析

RNAi transformation of Ah FAD2 gene and fatty acid analysis of transgenic seeds

在构建了以花生△12脂肪酸去饱和酶(FAD2)基因自身启动子驱动、有内含子间隔的该基因编码序列反向重复片段的RNA干扰载体的基础上,对上述干扰载体进行了农杆菌介导的花生胚小叶的遗传转化,以期特异调控花生籽粒中油酸、亚油酸含量。在2个受体品种中获得105株抗性再生植株,用两对引物对其进行了PCR扩增,其中32株初步证实为转基因植株。采用近红外分析仪对转基因T1籽粒油酸和亚油酸含量的检测结果表明,转基因株系内油酸/亚油酸比值的变异高于对照,多数转基因株系油酸亚油酸比值平均数也显著高于对照。

An RNAi vector for peanut FAD2 gene can be activated by its own promoter and containing its inverted repeat coding sequence fragments spliced by an intron. This gene was transformed into leaflet from germinating peanut seeds by the mediation of Agribacterium in order to manipulate the content of oleic acid and linoleic acid in the seeds specifically. A total of 105 plants tolerant to PPT were regenerated from two recipient cuhivars, in which 32 plants were confirmed by PCR to be tansgenic plants. Every seed produced by the transgenic plants was tested for its oleic and linoleic content by Infratec 1 255 Food＆Feed Analyzer. It was shown that the variation of oleic/linoleic ratio （O/L） within transgenic plants were greater than that within the wild type. T - test revealed that the average O/L values of seeds in most transgenic lines were significantly higher than that of the wild type,implying the efficiency of RNAi vector.