生物技术进展 ›› 2015, Vol. 5 ›› Issue (6): 436-440.DOI: 10.3969/j.issn.2095-2341.2015.06.06

• 研究论文 • 上一篇    下一篇

银柴胡离体培养与毛状根诱导技术初步研究

胡海英1,吴晓玲2   

  1. 1.宁夏大学科技处, 银川 750021;
    2.宁夏大学生命科学学院, 银川 750021
  • 收稿日期:2015-05-22 出版日期:2015-11-25 发布日期:2015-06-12
  • 作者简介:胡海英, 副教授,研究方向为植物生物技术。E-mail:haiying@nxu.edu.cn
  • 基金资助:

    宁夏自然科学研究基金项目(NZ13034)资助。

Studies on Isolated Culture and Hairy Roots Inducing of Stellaria dichotoma L. var. laceolata Bge

HU Hai-ying, WU Xiao-ling   

  1. 1.Science and Technology Department, Ningxia University, Yinchuan 750021, China;
    2.Life Science College, Ningxia University, Yinchuan 750021, China
  • Received:2015-05-22 Online:2015-11-25 Published:2015-06-12

摘要: 以银柴胡茎段为外植体,经消毒获得无菌再生材料后,筛选发根农杆菌介导毛状根诱导产生的最适条件。结果显示:最适的无菌消毒方法为:70%酒精浸5 s,0.1%升汞消毒3 min,获得了银柴胡离体培养材料;以发根农杆菌A4菌株介导的银柴胡毛状根诱导过程中,与叶片和不带腋芽茎段相比,带腋芽茎段为最适转化外植体,用OD600=0.8的菌液侵染茎段15 min,共培养3 d,800 mg/L头孢噻肟钠除菌,其诱导率及诱导密度最高,分别为100%和4.7,为最适诱导条件。研究结果说明在适合条件下,银柴胡带腋芽茎段适于诱导毛状根。

关键词: 银柴胡, 离体培养, 发根农杆菌, 毛状根, 诱导

Abstract: In this study, stems of Stellaria dichotoma L. var. laceolata Bge as explants  were sterilized, and sterile culture material were acquired. Then its hairy roots were induced by Agrobacterium rhizogenes A4 with Ri-plasmid. Results showed the method  that the explants was soaked in 70% alcohol for 5 s and in 0.1% HgCl2 for 3 min, was the optimal sterilized method, so sterile culture material were acquired. During the induction by Agrobacterium rhizogenes A4, comparing with the leaves and stems with no axillary bub taking the stems with axillary bud was the best explant for transformation. The induction rate and the density was 100% and 4.7, respectively, when the stems with axillary bud were infected for 15 min and co-cultured for 3 days with Agrobacterium rhizogenes (OD600=0.8), and sterilized by 800 mg/L cefotaxime sodium. So these were the optimized factors, and the stems with axillary bud were the optimized explants for transformation.

Key words: Stellaria dichotoma L. var. laceolata Bge, isolated culture, Agrobacterium rhizogenes, hairy roots, induction