Hairy root culture of Przewalskia tangutica for enhanced production of pharmaceutical tropane alkaloids
Hairy root culture of Przewalskia tangutica for enhanced production of pharmaceutical tropane alkaloids
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发表时间：
2010
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通讯作者：
Lan Xiaozhong;Q. Hong
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作者：
Lan Xiaozhong;Q. Hong
从唐古特高原普氏原羚种子萌发出无菌苗。用发根农杆菌A4菌株介导的遗传转化法，在乙酰丁香酮的诱导下，以叶片为起始外植体进行遗传转化。从叶片受伤部位诱导毛状根的频率为100%。将独立转化的快速生长的毛状根在MS培养基上进行继代培养，建立毛状根培养系。利用基因组DNA PCR技术检测了唐古特普氏原羚基因组中整合的生根基因rolB和rolC的存在。在MS液体培养基中培养5周后，毛状根产生的东莨菪碱和莨菪碱水平高于野生型根。东莨菪碱最佳生产线的东莨菪碱产量为0.68mg/g DW，莨菪碱最佳生产线的东莨菪碱产量为1.13mg/g DW。首次建立了唐古特毛状根培养体系，生产托品烷类生物碱。
The bacteria-free seedlings of Przewalskia tangutica were germinated from seeds. The leaves were used as initial explants for genetic transformation mediated by Agrobacterium rhizogenes strain A4, with the induction of Acetosyringone. The hairy roots were induced from the wounded sites of the leaves at the frequency of 100%. The independently transformed and fast growing hairy roots were subcultured in MS medium to establish the hairy root culture lines. Genomic DNA PCR analysis was applied to detect the presence of the rooting genes, including rolB and rolC that had integrated into the genome of Przewalskia tangutica. After 5-week culture in MS liquid medium, the hairy roots produced scopolamine and hyoscyamine at higher levels by comparison to the wild-type roots. The best scopolamine-production line produced 0.68 mg/g DW scopolamine, and the best hyoscyamineproduction line produced 1.13 mg/g DW hyoscyamine. It was the first time that hairy root cultures of P. tangutica were established to produce tropane alkaloids.