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园艺学报 ›› 2020, Vol. 47 ›› Issue (4): 635-642.doi: 10.16420/j.issn.0513-353x.2019-0695

• 研究论文 • 上一篇    下一篇

发根农杆菌介导的荔枝LcMYB1转化烟草叶片的研究

秦雅琪,胡桂兵,赵杰堂*   

  1. 华南农业大学园艺学院,亚热带农业生物资源保护与利用国家重点实验室,华南地区园艺作物生物学与种质创制农业部重点实验室,广东省荔枝工程技术中心,广州 510642
  • 出版日期:2020-04-25 发布日期:2020-04-25
  • 基金资助:
    国家自然科学基金项目(31872066);国家现代农业产业技术体系建设专项资金项目(CARS-32)

Studies on Agrobacterium rhizogenesis-mediated Transformation of LcMYB1 Gene into Tobacco Leaves

QIN Yaqi,HU Guibing,and ZHAO Jietang*   

  1. College of Horticulture,State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(South China),Ministry of Agriculture and Rural Affairs,Guangdong Litchi Engineering Research Center,South China Agricultural University,Guangzhou 510642,China
  • Online:2020-04-25 Published:2020-04-25

摘要: 利用荔枝果皮花青苷生物合成的关键调节基因LcMYB1为标记,构建植物表达载体pBA002-LcMYB1,电转法转入发根农杆菌A4菌株,通过叶盘法转化烟草K326。结果表明,烟草叶片侵染一周后长出白色的毛状根,约3周后部分毛状根逐渐变成红色,毛状根诱导率为100%,红色毛状根诱导率为19.9%。当筛选培养基中添加5 mg ? L-1 Basta,毛状根诱导率降低为33.3%,但红色毛状根诱导率提高到75%。高效液相色谱分析表明,红色毛状根中积累矢车菊素–3–葡萄糖苷和矢车菊素–3–芸香糖苷,PCR检测证实红色毛状根是由LcMYB1的转入引起的。实时荧光定量PCR表明转LcMYB1可以诱导烟草毛状根中花青苷生物合成相关的结构基因和调节基因表达。

关键词: 荔枝, 花青苷, MYB, 发根农杆菌, 烟草, 转基因, 毛状根

Abstract: A plant expression vector pBA002-LcMYB1 was constructed by using LcMYB1,a key regulator of anthocyanin biosynthesis in Litchi chinensis,and transferred into tobacco K326 mediated by Agrobacterium rhizogenes A4 strain. Results showed that white hairy roots grew one week after leaves infection,and some hairy roots gradually turned red about three weeks later. The induction rate of hairy roots was 100%,and that of red hairy roots was 19.9%. When 5 mg ? L-1 Basta was added to the screening medium,the induction rate of hairy roots decreased to 33.3%,but the induction rate of red hairy roots increased to 75%. High performance liquid chromatography analysis showed that cyanidin-3-glucoside and cyanidin-3-rutoside were accumulated in the red hairy roots. PCR analysis confirmed that the red hairy roots were caused by the over expression of LcMYB1 gene. Real-time quantitative PCR showed that LcMYB1 gene could induce the expression of structural and regulatory genes involved in anthocyanin biosynthetic in hairy roots of tobacco.

Key words: Litchi chinensis, anthocyanin, MYB, Agrobacterium rhizogenes, tobacco, transgenic, hairy root

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