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Agrobacterium-Mediated Genetic Transformation of the Medicinal Plant Veratrum dahuricum

文献类型： 外文期刊

作者： Ma, Rui 1 ; Yu, Zhijing 1 ; Cai, Qinan 1 ; Li, Haiyun 1 ; Dong, Yingshan 1 ; Oksman-Caldentey, Kirsi-Marja 2 ; Rischer 1 ;

作者机构： 1.Jilin Acad Agr Sci, Jilin Prov Key Lab Agr Biotechnol, Changchun 130033, Peoples R China

2.VTT Tech Res Ctr Finland Ltd, POB 1000, FI-02044 Espoo, Finland

关键词： Veratrum dahuricum; cyclopamine; Agrobacterium tumefaciens; transformation

期刊名称：PLANTS-BASEL （ 2020影响因子：3.935； ）

ISSN：

年卷期： 2020 年 9 卷 2 期

页码：

收录情况： SCI

摘要： Veratrum dahuricum L. (Liliaceae), a monocotyledonous species distributed throughout the Changbai mountains of Northeast China, is pharmaceutically important, due to the capacity to produce the anticancer drug cyclopamine. An efficient transformation system of Veratrum dahuricum mediated with Agrobacterium tumefaciens is presented. Murashige and Skoog (MS) medium containing 8 mg/L picloram was used to induce embryogenic calli from immature embryos with 56% efficiency. A. tumefaciens LBA4404 carrying the bar gene driven by the cauliflower mosaic virus 35S promoter was employed for embryogenic callus inoculation. A. tumefaciens cell density OD660 = 0.8 for inoculation, half an hour infection period, and three days of co-culture duration were found to be optimal for callus transformation. Phosphinothricin (PPT, 16 mg/L) was used as the selectable agent, and a transformation efficiency of 15% (transgenic plants/100 infected calli) was obtained. The transgenic nature of the regenerated plants was confirmed by PCR and Southern blot analysis, and expression of the bar gene was detected by RT-PCR and Quick PAT/bar strips. The steroid alkaloids cyclopamine, jervine, and veratramine were detected in transgenic plants, in non-transformed and control plants collected from natural sites. The transformation system constitutes a prerequisite for the production of the pharmaceutically important anticancer drug cyclopamine by metabolic engineering of Veratrum.

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