山银花的组培快繁和种质离体保存研究

Study on Rapid-propagation and in Vitro Preservation of Lonicera confusa DC

关键词:  组培快繁  离体保存  山银花  硝普钠

Key words:  tissue culture and rapid-proliferation  in vitro preservation  Lonicera confusa DC.  sodium nitroprusride

摘要:

以山银花(Lonicera confusa DC.)嫩茎段为外植体,研究不同培养基(MS、1/2MS、1/3MS、N68、White)和植物生长调节剂(6-BA、IBA、NAA、2,4-D、BR)对其诱导、分化和增殖的影响,建立山银花的快速繁殖再生体系,并探讨在温度15~20℃条件下,不同保存培养基(MS、1/2MS、1/4MS)对种质离体保存的影响。结果表明:山银花嫩茎段分化能力较强,在培养基MS+6-BA0.5mg·L^-1+IBA0.1mg·L^-1上培养7d腋芽开始萌发,15d时的诱导率达到90%;以N68为基本培养基,附加6-BA0.4mg·L^-1和IBA0.4mg·L^-1能较好地促进芽的伸长和增殖,避免叶片发黄脱落,每30d的增殖倍数为5.0倍;生根培养基以1/2MS+IBA0.2mg·L^-1最好,培养30d时生根率达95%;以单株芽段为离体保存材料,在1/2MS+蔗糖20g·L^-1培养基上培养30d后无菌条件下加入0.5mg·L^-1硝普钠溶液,继代间隔期可以延长至180d。

Abstract:

Using tender stems as explants, experiments were conducted to study the effects of media and exogenous hormones on inducement, differentiation, proliferation of Lonicera confusa DC., and to establish its regeneration system.Meanwhile, the impacts of media on in vitro preservation when the temperature was 15~20℃ were investigated.According to the results, the axillary buds could be well induced on the medium MS+6-BA0.5 mg·L^-1+ IBA0.1 mg·L^-1, on which they began to bourgeon after 7 days, with 90% inducement rate after 15 days.On N68 medium with 6-BA0.4mg·L^-1 and IBA0.4mg·L^-1, the propagation coefficient was 5.0 times every 30 days, and elongaton and proliferation of buds were promoted while preventing their leaves from etiolating and falling off.The ideal rooting medium was 1/2MS+IBA0.2mg·L^-1, on which rooting rates were 95% after 30 dayes.Being cultured on the preservation medium 1/2MS+sucrose 20 g·L^-1, 0.5mg·L^-1 sodium nitroprusride was added after 30 days in asepsis condition, the single caudexs could be cultivated for 180 days without being transfered.