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。
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% HgC12 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.
出处
《生物技术进展》
2015年第6期436-440,I0003,共6页
Current Biotechnology
基金
宁夏自然科学研究基金项目(NZ13034)资助
关键词
银柴胡
离体培养
发根农杆菌
毛状根
诱导
Stellaria dichotoma L. var. laceolata Bge
isolated culture
Agrobacterium rhizogenes
hairy roots
induction
