火龙果离体培养茎段灭菌及芽诱导增殖研究

Study on Stem Disinfection,Bud Induction and Proliferation of Hylocereus undatus in Vitro Culture

【目的】探讨火龙果茎段灭菌及芽诱导增殖技术,为研发火龙果组织培养技术提供支撑。【方法】以桂红龙1号火龙果茎段为外植体,研究不同因素对火龙果茎段灭菌及芽诱导增殖过程的影响。【结果】茎段大小是影响灭菌效果的主要因素,其次是灭菌时间,茎段老熟程度的影响相对较小。灭菌时间对芽诱导的影响最大,6-BA浓度次之,茎段老熟程度和茎段大小对芽诱导影响较小,且差别不大。6-BA 4.0 mg/L时芽诱导效果最佳。6-BA浓度越高,芽增殖倍数越高,但当6-BA达到4.0 mg/L时,芽出现玻璃化现象。IAA的芽增殖效果优于NAA,IAA为0.1 mg/L时芽增殖效果较好。在培养基中添加PP333对抑制玻璃化促进火龙果组培苗继代增殖的效果优于AC。【结论】适宜茎段灭菌及芽诱导的方案为:选取生长1个月左右的健康茎段,切成含1个芽眼的小段,用0.1%Hg Cl_2溶液灭菌10 min,将灭菌的茎段接种至MS+6-BA 4.0 mg/L培养基。适宜芽继代增殖的培养基配方为:MS+6-BA 3.0 mg/L+IAA 0.1 mg/L+PP333 1.0 mg/L。

[ Objective]Hylocereus undatus stem disinfection, bud induction and proliferation were explored during tissue culture to provide technical support for mass seedling production. [Method]Taken stem ofHylocereus undatus cuhivar Guihonglong No. 1 as the explants, the effects of different factors on stem disinfection, bud inducement and proliferation were studied. [ Result] Stem size was the main factor affect- ing the pollution rate of stem disinfection, followed by the effect of sterilization time and stem mature degrees. The sterilization time had the greatest influence on the bud induction, followed by the 6-BA concentration. Both the stem mature degrees and stem size had little effect on bud induction. 6-BA 4.0 mg/L had the best induction effect. The higher concentration of 6-BA showed the higher multiplication ratio of buds, but when 6-BA concentration reached 4.0 mg/L, the buds appeared vitrification. The effect of IAA was better than that of NAA, and the effect of bud proliferation was better when IAA was 0.1 mg/L. PF333 was better than AC in inhibiting vitrification and promoted subcul- ture multiplication of Hylocereus undatus. [ Conclusion ] The optimal treatment for stem disinfection and bud induction was suggested as：sliced the health stems of about 1 months into pieces with single bud, and disinfected them with 0.1% HgC12 solution, and inoculated the explants in medium of MS ＋ 6-BA 4.0 mg/L. The optimal medium for bud proliferation was MS ＋6-BA 3.0 mg/L ＋ IAA 0. 1 mg/L ＋ PP333 1.0 mg/L.