西葫芦大孢子离体培养再生植株技术优化研究

西葫芦大孢子离体培养过程中,培养体玻璃化是再生植株形成的主要障碍之一。为保障西葫芦大孢子培养过程中胚状体的正常产生,减少培养体玻璃化对再生植株形成的影响,提高试管苗成活率。本试验通过4种培养基诱导胚状体形成筛选优良诱导培养基,通过对组织培养过程中环境温度、湿度、GA浓度等因子调整来降低培养体玻璃化的发生。结果表明：4种培养基均能不同程度诱导胚状体形成,其中D号B5 40g/LSuc 8g/LAgar 0.5mg/LNAA 0.05mg/LTDZ 30mg/LAgNO3培养基最佳(诱导出胚达96％),且直接生成76％不定芽；在培养温度为25℃,湿度85％,生根培养基GA浓度0.5mg/L条件下西葫芦大孢子培养体玻璃化发生率最低。这项技术在前人基础上完善、优化、建立了最佳的西葫芦未受精子房再生植株技术体系,为后续西葫芦的研究奠定基础。

Improving Technology of Megaspore Culture in Vitro and Plantlet Regeneration in Cucurbita pepo

The vitrification phenomenon of culturing body is one of the main obstacles to regenerated plantlet formation in the process of in vitro culture of megaspore zucchini (Cucurbita pepo). The aim of the research is to guarantee normal production of the embryoid, minimize the impact of vitrification phenomenon on plantlet regeneration from megaspore and enhance the survival rate of test-tube plantlets. In the experiment, 4 kinds of media-induced somatic embryogenesis were selected to induce the formation of excellent induction medium. And the environmental temperature, humidity and concentration of GA were adjusted to reduce the occurrence of vitrification shoots of culturing body. The results showed that the four kinds of different levels of culture medium could induce the formation of embryoid bodies. One of the best was No. D: B5 40 g/L Suc 8 g/L Agar 0.5 mg/L NAA 0.05 mg/L TDZ 30 mg/L AgNO3. Meanwhile, embryo induction was up to 96% and 76% adventitious bud was generated directly. In the condition of 25℃ , 85% humidity, and 0.5% rooting medium GA concentration, the vitrification shoots rate of culturing body was the lowest. On the basis of previous researches, the experiment optimized and established the best zucchini plant regeneration unfertilized room technology system, provided a reference for further study on zucchini.