基于间歇浸没式生物反应器的荔浦芋组培快繁体系优化

目的]优化基于间歇浸没式生物反应器系统(TIBs)的荔浦芋组培快繁体系,为荔浦芋脱毒组培苗的工厂化、自动化生产提供技术支持.方法]以荔浦芋新品种桂芋2号茎尖分生组织脱毒诱导获得的不定芽为外植体,利用TIBs培养系统筛选出适合组培苗增殖及生根的最佳激素组合,并分析不同继代材料、接种密度及浸没间歇频率对组培苗增殖和生根效果的影响.结果]利用TIBs培养系统可有效提高荔浦芋组培苗的增殖效果,增殖倍数达28.96倍,约是传统固体培养方法(2.87倍)的10倍,且组培苗的株高和生根数均极显著高于传统固体培养植株(P<0.01).在TIBs培养系统中,含4.00 mg/L 6-苄氨基嘌呤(6-BA)+0.05 mg/L萘乙酸(NAA)的培养基最适合荔浦芋组培苗增殖和生长,其组培苗增殖倍数为32.04倍.当接种材料为第4代荔浦芋继代材料、接种密度为10株/L、浸没间歇频率为5 min/6 h时,最有利于组培苗增殖和生长,增殖倍数均在30.00倍以上,可缩短萌芽时间,组培苗长势良好,且培养基污染率为0.结论]利用TIBs培养系统对荔浦芋继代材料进行高效快繁具有可行性,可为实现及推动荔浦芋健康种苗繁育的工厂化生产提供技术支持.

Optimization of Lipu taro tissue culture rapid propagation in temporary immersion bioreactors system

Objective]The rapid propagation system of Lipu taro was optimized by using temporary immersion biore-actors system(TIBs)in order to provide technical supports for industrialized and automated production of its virus-free plantlets.Method]The adventitious buds induced by meristem of Lipu taro variety Guiyu 2 was used as explants,the suitable hormone combination for multiplication and rooting of adventitious buds was screened by using TIBs,and the effects of different subculture materials,inoculated densities and intermittent frequencies of TIBs on the multiplication and rooting were analyzed.Result]In TIBs,the multiplication effect of Lipu taro plantlets was enhanced effectively,and the multiplication rate of buds reached 28.96 times,which was as 10 times as that of traditional solid culture(2.87 times);and both of plantlet height and rooting number of induced plantlets were extremely higher than those of traditional solid culture(P<0.01). The hormone combination of 4.00 mg/L 6-BA+0.05 mg/L NAA added in culture medium was the best for multiplication and growth of Lipu taro plantlets in TIBs,and the multiplication rate of plantlet reached 32.04 times. When the 4th sub-cultured plantlet was used as explants,10 plantlets per liter of inoculation densities and 5 min per 6 hours of immersion frequency were found to be the best for growth and multiplication of plantlets,the multiplication rate reached more than 30.00 times,and could shorten the germination of plantlets with well growth vigor in TIBs,and the contamination rate of culture medium was 0.Conclusion]The results showed that it is of highly practicable to use TIBs for rapid propagation of Lipu taro sub-culture plantlets,this will provide important technical supports for implementing and promoting large-scale industrialized production of Lipu taro healthy seedling propagation.