木豆高频不定芽再生及体细胞遗传变异的分子检测

Efficient Regeneration from Cotyledonary Nodes of Pigeon Pea Via Organogenesis and Genetic Fidelity of Adventitious Shoots as Revealed by ISSR Markers

以木豆子叶节为外植体,建立高频不定芽再生体系,并对继代丛芽的遗传变异进行分子检测。结果表明:子叶节不定芽再生的最佳培养基为EC6附加4.0mg/LBA和0.1mg/LIBA;丛芽增殖培养以EC6附加4.5mg/LBA、0.1mg/LIBA和0.4mg/LKT效果最佳。同时,随机挑选继代第5代的20个丛芽提取基因组DNA,采用45个ISSR引物对基因组DNA进行分子检测,结果显示,在所用引物范围内均未检测到变异。

With cotyledonary nodes of pigeon pea（Cajanus cajan） as explant to establish efficient regeneration system and detect genetic variation of adventious buds of Pigeon pea（Cajanus cajan） with molecule biology method.The result showed that the EC6 supplemented with both 4.0 mg /L BA and 0.1 mg /L IBA was the best medium for cotyledonary nodes regeneration efficiently.Bud regeneration cultivation was the best in EC6 medium supplement with the combination of 4.5 mg /L BA,0.1 mg /L IBA and 0.4 mg /L KT.To investigate the genetic stability of in vitro shoots,20 in vitro shoots sellected randomly from the 5th cycle of transfer were employed to carried out PCR using 45 ISSR primers.The result showed no variation was detected among all primers.