摘要
利用已构建载体530Shiva,转化马铃薯普通栽培种甘农薯1号、澳布、HLS和夏波蒂的试管苗及块茎盘的研究。结果表明,以Kan50mg·L-1和100mg·L-1作为筛选转基因细胞和植株的剂量。对于发根农杆菌侵染马铃薯试管苗应采用新鲜菌落直接涂抹伤口进行侵染,这种方法诱导生根率高且易于操作,根诱导率、成愈率及分化率远高于浸泡法;发根农杆菌转化试管苗,后期愈伤组织分化产生5个幼苗。发根农杆菌菌液侵染甘农薯1号和夏波蒂两品种的块茎盘,7d后就有很小的毛状根突起产生,2周后即有大量毛状根产生,3周后毛状根产生量有所减少,产生的毛状根3周时其成愈率分别为31.0%和27.0%,愈伤组织分化成苗比较困难,分化出3个幼苗,分化率分别为6.45%和3.70%。转化材料进行冠瘿碱检测表明,分化苗均为胭脂碱阳性反应。环腐病菌接种试管苗结果表明,分化植株与对照相比抗性明显增强。
The potato cvs. Gannongshul, Aobu, HLS and Shepody were transformed using the bacterium strain containing Ri plasmid 530 shiva with Shival gene. In this research, the 50 mg·L^-1 and 100mg·L^-1 were used as the selection dosages for selecting the transgenic plants（Kan^R）. The results showed that the wounded potato tube-seedling should be directly transformed using fresh colony. This method was easy to operate and the hair-root, calli and regenerated plantlets were higher than dipping method. Five plantets were regenerated using this method. When dipping method was used to transform the tuber-disc of Grannongshul and Shepogy, very small hair-root was formed aftter 7 days of inoculation, and plentiful hair-roots was produced aftter 2 weeks. However, the production of hair-root was reduced aftter 3 weeks. The ratio of calli was 31.0% and 27.0%, respectively, for these two cuhivars. Three plantlets were regenerated, the ratio of regeneration being 6.45% and 3.70%, respec-tively. The transformed material contained the opine when tested. Furthermore, the transgenic test-tube plantlets were proved to have resistance to potato ring-rot disease.
出处
《中国马铃薯》
2007年第6期341-345,共5页
Chinese Potato Journal
关键词
发根农杆菌
Shival
转化
马铃薯
Agrobacterium rhizogenes
Shival
transformation
Solanum tuberosum
