不同处理对苜蓿小孢子离体培养存活率的影响

Effects of Survival Rate of Alfalfa Microspore Culture by Different Treatments

【目的】探讨不同处理对苜蓿小孢子培养的影响，为苜蓿游离小孢子培养体系的建立提供依据，也为今后苜蓿单倍体育种奠定基础。【方法】以杂花苜蓿新牧1号花蕾为材料进行小孢子离体培养，探讨预处理、培养基类型及接种密度对苜蓿小孢子离体培养前期存活率的影响。【结果】在预处理实验中，4℃低温处理24h存活率最高，达23．5％；33℃高温处理48h后转入25℃暗培养，小孢子存活率最高，达26．58％；按1．5×10^5个／mL的密度接种，小孢子存活率最高，达28．2％；小孢子在B5液体培养基中培养优于其它培养基，存活率达30．1％。【结论】材料在4℃低温下处理24h，将游离小孢子按1．5×10^5个／mL的接种密度接种于B5液体培养基中，置于33℃高温下处理48h后转入25℃恒温培养箱中暗培养，25d后胚细胞诱导率最高，达30．1％。

[ Objective] The purpose of this project is to study the effects of survival rate of alfalfa microspore culture in initial stage by different treatments in order to provide the theoretical foundation for the establishment of alfalfa microspore culture system. [ Method ] The experiment was conducted by using alfalfa buds（M, varia Martyn cv. Xinmu No. 1 ）as material. The effects of survival rate of alfalfa microspore culture in initial stage by different pretreatment, medium and inoculum density were studied. [ Result ] The results indicate that it had the highest survival rate （ 23.5% ） when pretreated 24 h at 4℃ ; And in the high temperature pretreatment, the 48 h pretreatment at 33% which then continued to train under the dark culture conditions of 25℃ had better effect, the survival rate reached 26.58% ; In the trims of screening medium, the effect of B5 medium was better than others, and in the screening density test, conditions of this density had higher survival rate when it was 1.5 × 10^5/mL, the survival rate reached 30.1%. [ Conclusion] This test has obtained the following conclusions：After the materials at 4℃ for 24 h, isolated microspores were vaccinated in B5 medium with the density of 1.5 × 10^5/mL. and they were placed in the high temperature of 33℃ for 48 h and then transferred to constant temperature of 25℃ under the dark. Embryo cells were induced the highest after 25 days incubation to 30.1%.