文献类型： 外文期刊

作者： Mi, Yaolei ¹ ; Zhu, Zhihui ³ ; Qian, Guangtao ³ ; Li, Yu ⁴ ; Meng, Xiangxiao ¹ ; Xue, Jianping ³ ; Chen, Qingfu ⁵ ; Sun, Wei ¹ ; Shi, Yuhua ¹ ;

作者机构： 1.China Acad Chinese Med Sci, Inst Chinese Mat Med, Key Lab Beijing Identificat & Safety Evaluat Chin, Beijing, Peoples R China

2.China Acad Chinese Med Sci, Artemisinin Res Ctr, Beijing, Peoples R China

3.Huaibei Normal Univ, Coll Life Sci, Huaibei, Anhui, Peoples R China

4.Sichuan Acad Agr Sci, Econ Crop Res Inst, Chengdu, Sichuan, Peoples R China

5.Guizhou Normal Univ, Res Ctr Buckwheat Ind Technol, Guiyang, Guizhou, Peoples R China

关键词： Genetics; Issue 157; Tartary buckwheat; hairy roots; genetic transformation; GFP; Agrobacterium rhizogenes; secondary metabolite; gene function

期刊名称：JOVE-JOURNAL OF VISUALIZED EXPERIMENTS （ 影响因子：1.355； 五年影响因子：1.696 ）

ISSN： 1940-087X

年卷期： 2020 年 157 期

页码：

收录情况： SCI

摘要： Tartary buckwheat (TB) [Fagopyrum tataricum (L.) Gaertn] possesses various biological and pharmacological activities because it contains abundant secondary metabolites such as flavonoids, especially rutin. Agrobacterium rhizogenes have been gradually used worldwide to induce hairy roots in medicinal plants to investigate gene functions and increase the yield of secondary metabolites. In this study, we have described a detailed method to generate A. rhizogenes-mediated hairy roots in TB. Cotyledons and hypocotyledonary axis at 7-10 days were selected as explants and infected with A. rhizogenes carrying a binary vector, which induced adventitious hairy roots that appeared after 1 week. The generated hairy root transformation was identified based on morphology, resistance selection (kanamycin), and reporter gene expression (green fluorescent protein). Subsequently, the transformed hairy roots were self-propagated as required. Meanwhile, a myeloblastosis (MYB) transcription factor, FtMYB116, was transformed into the TB genome using the A. rhizogenes-mediated hairy roots to verify the role of FtMYB116 in synthesizing flavonoids. The results showed that the expression of flavonoid-related genes and the yield of flavonoid compounds (rutin and quercetin) were significantly (p < 0.01) promoted by FtMYB116, indicating that A. rhizogenes-mediated hairy roots can be used as an effective alternative tool to investigate gene functions and the production of secondary metabolites. The detailed step-by-step protocol described in this study for generating hairy roots can be adopted for any genetic transformation or other medicinal plants after adjustment.