甘蔗茎尖胚状体脱毒苗快繁技术研究

Sugarcane Micropropagation through Shoot-tip Embryogenesis

以甘蔗新台糖22号为材料,比较茎尖胚状体、茎尖与腋芽3种分化成苗繁育方法在不同时期接种、不同激素水平下的组培苗增殖速度、苗素质及脱毒效果。试验结果表明：组培苗繁殖速度以茎尖胚状体分化苗最快,增殖5代后扩繁2589倍,茎尖297倍,腋芽104倍,培养基以6-BA 1.5mg/L＋NAA 0.01-0.1mg/L增殖效果最好;组培苗质量与繁殖速度相反;出现不正常生长的苗类型有白化苗、细弱小苗、玻璃化苗、疯长苗4种,茎尖胚状体苗发生率1.77%,茎尖苗1.56%,腋芽苗0.31%;不同处理间组培苗生根及移栽成活率差异不显著,生根率茎尖胚状体苗75.3%、茎尖苗76.9%、腋芽苗76.6%,移栽成活率茎尖胚状体苗94.8%、茎尖苗95.4%、腋芽苗95.1%,生根培养基以NAA7.5mg/L＋ABA 2.5mg/L最好;去除RSD、花叶病方面,以茎尖胚状体苗最好,RSD去除率95%、花叶病去除率100%,茎尖苗RSD去除率70%、花叶病75%,腋芽苗未能去除RSD、花叶病。应用茎尖胚性细胞再生植株,脱毒效果好,繁殖速度快,可克服目前脱毒苗生产中试管苗扩繁量小、成本高的难题。

Sugarcane ROC22 was selected as the experimental material;multiplication rate,plantlet＇s quality and effect of virus-free were compared between different time for inoculating and different concentration of hormones of sugarcane plantlets from embryogenesis,shoot tips and apical buds.After 5 subcultures on MS ＋ 6-BA 1.5mg/L ＋ NAA 0.01-0.1 mg/L medium generated 2589 plantlets from embryogenesis,297 plantlets from shoot tips and 104 plantlets from apical bud.Quality of plantlets opposite to multiplication rate,some abnormal plantlet（white,slim,vitrification and crazy-growing plantlet） occurred in the three methods（incidence of embryogenesis,shoot tips and apical bud was 1.77%,1.56% and 0.31%,respectively）.There was no significant difference between the different treatments of rooting rate and survival rate,the rooting rate of embryogenesis,shoot tips,and apical buds was 75.3%,76.9% and 76.6%,respectively;the survival rate of embryogenesis,shoot tips and apical buds was 94.8%,95.4% and 95.1%,respectively.MS＋NAA 7.5mg/L＋ABA 2.5mg/L was better for rooting.Plantlets generated from shoot tips embryogenesis was better for wiping off RSD and mosaics disease.The method of shoot tips embryogenesis for generating sugarcane plantlets is good for wiping off virus and enhancing the multiplication rate,and is better for overcome the problems of lower multiplication rate and high cost in sugarcane tissue culture.