Raise up of Scopolamine in Hairy Roots Via Agrobacterium rhizogenes ATCC15834 as Compared with Untransformed Roots in Atropa komarovii.

Ofelia Banihashemi, Ramazan-Ali Khavari-Nejad, Narguess Yassa, Farzaneh Najafi
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引用次数: 1

Abstract

Atropa komarovii generates tropane alkaloids and three other compounds such as hyoscyamine. Racemate atropine and scopolamine (hyoscine) are the main alkaloids with anticholinergic, antispasmodic, and sedative agents. A proficient convention has been reported for the formation of transgenic Atropa komarovii by the use of Agrobacterium rhizogenes. Root culture, by utilizing leaves explants was contaminated by Agrobacterium rhizogenes ATCC 15834, a strain with the paired vector. The hairy roots after contamination for three weeks were specifically shaped from the cut edges of the leaves. The PCR intensification demonstrated that rol B genes of Ri plasmid of Agrobacterium rhizogenes were coordinated and communicated into the genome of the changed hairy roots. Examination of HPLC revealed that hairy roots can produce scopolamine and hyoscyamine and it was appeared that scopolamine content was essentially expanded in changed roots and hyoscyamine was extremely expanded in non-transgenic roots. According to the results, it was perceived that the scopolamine content in hairy roots was raised significantly compared to the control roots. It was evidenced that hairy roots gather a great number of metabolites that have a commercial significance. Thus, later on we can enhance efficiency for example by building up the biosynthetic route overexpression of gene codifying enzymes in the metabolic route for expanding valuable secondary metabolites in the plant cures.

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发根农杆菌ATCC15834对黄芪毛状根中东莨菪碱含量的提高与未转化根的比较
Atropa komarova产生莨菪烷生物碱和其他三种化合物,如莨菪碱。外消旋体阿托品和东莨菪碱是具有抗胆碱能、抗痉挛和镇静作用的主要生物碱。一个熟练的惯例已经报道了形成转基因阿托罗夫利用根生农杆菌。利用叶片外植体进行根培养,感染了具有配对载体的根生农杆菌ATCC 15834。污染三周后的毛状根是由叶子的切割边缘形成的。PCR强化结果表明,根农杆菌Ri质粒的rol B基因被协调并传递到变异毛状根的基因组中。HPLC检测结果表明,毛状根可产生东莨菪碱和山莨菪碱,并且在转基因根中东莨菪碱含量基本增加,而在非转基因根中山莨菪碱含量增加。结果表明,与对照根相比,毛状根中东莨菪碱含量显著提高。有证据表明,毛状根收集了大量具有商业意义的代谢物。因此,以后我们可以提高效率,例如通过建立生物合成途径,在代谢途径中过度表达基因编码酶,以扩大植物药材中有价值的次生代谢物。
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