菜心组织培养技术初探

Preliminary Study on Tissue Culture Technique in Brassica campestris L. ssp. chinensis var. utilis

为建立菜心(Brassica campestris ssp.chinensis var.utilis)的快繁技术体系,以花药和子叶-子叶柄为外植体进行组织培养研究。结果表明,花药培养以选取未开放的花蕾为宜,且花柱略高于花瓣,此时小孢子多数处于单核靠边期。菜心花粉的萌发率不高,且秋冬季的花粉比夏季的萌发率高。菜心花药愈伤组织诱导培养基为:MS+1.0 mg L–1 KT+1.0 mg L–1 2,4-D+3%糖+6 g L–1琼脂+8%椰乳,不定芽诱导培养基为:MS+2.0 mg L–1 6-BA+0.5 mg L–1 NAA+1.0 g L–1活性炭+2%糖+6 g L–1琼脂或MS+2.0 mg L–1 ZT+0.5 mg L–1 IAA+0.5 g L–1 AgNO3+1.0 g L–1活性炭+2%糖+6 g L–1琼脂。花药培养的不定芽诱导率为36.7%,不定芽培养出现褐化现象,不能形成再生植株;而以子叶-子叶柄为外植体培养获得的植株再生率可达80%。

In order to establish rapid propagation system of Brassica campestris ssp. chinensis var. utilis, the anthers as explants were in vitro cultured. The results showed that the anthers should be selected from unopened buds, which stigma was slightly higher than petal, and most of microspores were at uninucleate stage. The pollen germination rate was not high, and that in autumn and winter was higher than that in summer. The callus induction medium for anthers was MS＋1.0 mg L-1 KT＋1.0 mg L-1 2,4-D＋3%sugar＋6 g L-1 agar＋8%coconut milk （pH=5.8）. The adventitious bud differentiation medium was MS＋2.0 mg L-1 6-BA＋0.5 mg L-1 NAA＋1.0 g L-1 active carbon＋2%sugar＋6 g L-1 agar or MS＋2.0 mg L-1 ZT＋0.5 mg L-1 IAA＋0.5 g L-1 AgNO3＋1.0 g L-1 active carbon＋2%sugar＋6 g L-1 agar （pH=5.8）. The adventitious bud rate inducted from anthers was 36.7%, and the regeneration plantlet rate was low owing to adventitious buds browning, while the regeneration plantlet rate reached to 80%induced from cotyledon or petioles.