南瓜种子无菌苗培养技术研究

Culture Technology for Aseptic Seedling of Pumpkin

以南瓜"密本"种子为试材,研究种皮处理和种膜处理方式、催芽、培养基琼脂和蔗糖质量浓度对组织培养种子萌发及无菌苗生长情况的影响,分时段分析了各处理种子胚根萌发率、子叶变绿比率、侧根发生率、子叶展开率、第1真叶发生率,测量无菌苗真叶长、真叶宽、茎粗、节间长、植株高度、生长点高度等差异,筛选南瓜无菌苗最佳培养体系,以研究南瓜种子用组织培养方法培育无菌壮苗技术的最佳条件,为南瓜扩繁和遗传转化育种提供支持。结果表明:南瓜种子获取无菌苗的较好处理方式是将种子剥壳、留种膜,琼脂浓度5g·L^(-1)时,无菌苗真叶的分化最早,但无菌苗逐渐出现玻璃化;琼脂浓度6g·L^(-1)或8g·L^(-1)时,无菌苗的综合性状最优;培养基不加蔗糖,植株萌发期和幼苗前期生长与发育最快,而添加蔗糖30g·L^(-1),植株后期营养生长最旺盛。试验表明,对南瓜无菌种子培养时,将种子剥壳、留种膜,以培养基MS+30g·L^(-1)蔗糖+6g·L^(-1)琼脂为条件最适合培养南瓜无菌苗。

The‘Miben＇pumpkin seeds were used as materials,the influence of seed skin treatment,the type of membrane treatment,pregermination,the agar and sucrose concentration of culture medium on germination and tissue culture seedling were studied,for which a serious of indices such as radicle rate,lateral roots rate,ratio of cotyledon turning green,cotyledon expansion rate and first main leaf rate were analyzed,and differences of leaf blade length,leaf blade width,leaf blade coarseness,internode length,plant height and growing point height were measured to obtain the optimum cultivation system of pumpkin sterile seedling.This best condition of aseptic tissue culture pumpkin seedling cultivation was researched,to provide support for pumpkin breeding.The results showed that,the better way for pumpkin seeds to get aseptic seedlings was to peel the seed shell and preserve seed membrane,the true leaf differentiation appeared the earliest when agar concentration was5 g·L^-1,but the plants gradually appeared vitrified;the comprehensive properties of the sterile seedlings were optimal when agar concentration was 6 g·L^-1 or 8 g·L^-1;plant germination as well as seedling early growth and development went the fastest when medium was not added with sugar,plant late vegetative growth was the most exuberant when added with 30 g·L^-1 sugar.The influence factors of aseptic culture of pumpkin seedlings were discussed,among which pumpkin seeds were peeled with membrane intact and sown in MS medium with 6 g·L^-1 agar and 30 g·L^-1 sugar,to provide support for breeding pumpkin propagation and genetic transformation.