野葛毛状根离体培养与异黄酮生产

Pueraria lobata Hairy Root Culture in vitro and Isoflavone Production

用发根农杆菌 (Agrobacteriumrhizogenes)R16 0 1菌株感染野葛 (Puerarialobata (Willd .)Ohwi)叶片外植体后获得的毛状根 ,经连续 4次继代培养 ,鲜重增加到最初的 6 .2倍。pH为 5 .5的 1/ 2SH液体培养基较适合野葛毛状根的生长。与自然根相比 ,毛状根在无激素的 1/ 2SH液体培养基中离体培养 2 0d后 ,鲜重增加 2 2 .6倍 ,释放入培养液中的异黄酮量增加 10 .6倍。毛状根生产异黄酮的量略高于自然根 ,且根中含量明显高于愈伤组织及茎中异黄酮的含量。

Hairy roots were obtained from leaf explants of Pueraria lobata (Willd.) Ohwi after inoculation with Agrobacterium rhizogenes R1601 strain. The increment of fresh weight of hairy roots raised 5.2 times after the fourth in vitro subculture compared with the initial data(Fig.1). The optimum of culture condition for the growth of P. lobata hairy roots was observed on 1/2 SH medium with pH 5.5(Figs.2 and 3). Compared with the natural roots, the fresh weight of hairy roots suspended in hormone-free SH medium 20 days in vitro increased 22.6 times and the isoflavone content in culture medium increased 10.6-fold(Figs.4 and 7). The isoflavone content in the hairy roots was little higher than that in the natural roots and significantly higher than that in the calli and stems(Fig.6). The volume of the medium also affected the growth of hairy roots in vitro and the release of isoflavones(Figs.5 and 8).