摘要
利用发根农杆菌A4和R1000感染四倍体菘蓝子叶,毛状根诱导频率和得根率分别达72.5%和27%以上。A4诱导四倍体菘蓝毛状根优于R1000,光照对毛状根的诱导起着重要作用,外源激素IAA能明显地促进毛状根的诱导作用。rolB、rolC基因的PCR分析表明,Ri质粒中的T-DNA片段已整合到毛状根细胞基因组中。毛状根能在不含激素的MS固体培养基上分化出不定芽,6-BA能促进毛状根不定芽的诱导;NAA促进不定芽生根,再生完整植株。
Hairy root clones of Isatis indigotica Fort. transformed with Agrobacterium rhizogene strains A4 and R1000 were established. A4 had higher root-inducing capability than R1000. The percentage root induction by A4 was 72.5% and average root number per cotyledon was over 27. Both IAA and light had significant effects on root-inducing effeciency. For example, the percentage of root induction could be enhanced with MS medium supplemented with IAA of 0. 5mg/L and no root occurred without light. The presence of the rolB and rolC genes in the hairy roots was confirmed by PCR analysis. The positive result obtained in the amplication of the rolB and rolC in hairy roots could be attributed to successful transformation rather than to residual Agrobacterium. These hairy roots grew well on MS agar medium or liquid medium without growth regulator for over one year. Four hairy root clones, namely TRA1, TRA2 ,TRA3 and TRA4, were established They grew vigorously and produced numerous lateral branches. Twenty one buds from 30 root segment cultures were obtained directly from hairy roots after subcuhuring on medium lacking growth regulator for 90 days. 6-BA had significant effect on bud differentiation and thus the optimized medium for bud differentiation was MS ＋ NAA1.0mg/L ＋ 6 - BA 0. 5mg/L. The buds were rooted on root induction medium supplemented with NAA and developed into regenerated plants.
出处
《作物杂志》
CSCD
北大核心
2008年第5期31-35,共5页
Crops
关键词
四倍体菘蓝
毛状根
植株再生
Autotetraploid Isatis indigotica
Hairy root
Plantlet regeneration
