油松花粉离体萌发培养基及花粉贮藏条件研究

Medium for pollen germination of Pinus tabulaeformis in vitro and its pollen storage conditions

花粉生活力对油松杂交育种至关重要。为筛选油松花粉离体萌发培养基,探讨不同温度贮藏下花粉的生活力,采用正交试验设计,研究不同水平蔗糖、硼酸和pH值对油松花粉离体萌发的影响;测定不同温度贮藏下油松花粉萌发率随时间的动态变化。结果表明:油松新鲜花粉在蔗糖100 g·L^(-1)、硼酸0.10 g·L^(-1)、琼脂8 g·L^(-1)、pH值6.5的培养基上萌发效果最好,萌发率高达93.12%。不同贮藏温度下油松花粉生活力存在明显差异,-80℃贮藏15 d和30 d的花粉萌发率分别为92.93%和91.27%,萌发率下降不显著;-20℃和4℃贮藏15 d,花粉萌发率下降不显著,但贮藏30 d的花粉萌发率与初始萌发率相比均极显著下降,萌发率分别为88.65%和86.84%;常温25℃贮藏15 d、30 d的花粉萌发率分别为90.27%、82.66%,与初始萌发率均达到极显著差异;由此可见,贮藏温度越低,越有利于花粉生活力的保持。此研究可为油松花粉生活力检测提供一种可靠方法,并为花粉贮藏提供参考依据。

Pollen viability is of great importance for Pinus tabulaeformis cross breeding.In order to select suitable medium for pollen germination of P.tabulaeformis in vitro and understand the pollen viability under different storage temperature,the effects of different levels of sucrose,boric acid and pH on the pollen germination of P.tabulaeformis in vitro were studied by orthogonal experiments.At the same time,the dynamic changes of germination rate of P.tabulaeformis pollen stored at different temperature were measured using optimized medium.The results indicated that germination rate of fresh pollen in vitro was up to 93.12%when placed in the optimal medium containing 100 g·L^(-1) sucrose,0.10 g·L^(-1) boric acid,8 g·L^(-1) agar and pH 6.5.The pollen viability of P.tabulaeformis under different storage temperature had obviously differences,when the pollen were stored at -80℃ for 15 d and 30 d,their germination rates were 92.93%and91.27%,respectively,which had no significant decrease.When stored at -20℃ and 4℃ for 15 d,the pollen germination rate had no significant decrease,however,the germination rates of pollen stored 30 d declined significantly compared with their initial germination rate,which were 88.65%and 86.84%,respectively.When the pollen were stored at 25℃ for 15 d and 30 d,the germination rates were 90.27%and 82.66%respectively,which had significant differences from initial germination rate.It could be concluded that the lower the storage temperature was,the better viability the pollen maintained.The present study could provide a reliable method for pollen viability measurement of P.tabulaeformis and supply reference basis for pollen storage.