不同柳树花粉离体培养及活力测定方法比较研究

Comparative study on in vitro pollen germination and viability evaluation of different willow species

以6个不同种柳树亲本及杂交子代个体为试验材料,采集其花粉进行离体培养以确定最佳培养条件,同时比较离体培养法、蓝墨水染色法、I2-KI染色法和TTC染色法4种花粉活力测定方法,以期探索适合柳树不同种个体材料的花粉活力快速检测体系。结果表明:离体培养基中蔗糖、H_3BO_3和Ca(NO_3)_2的质量浓度配比因柳树个体不同而略有差异;不同培养时间对柳树花粉活力影响显著,适宜培养时间为10—12 h;光照条件对离体培养影响不显著;不同个体最适培养温度均为25℃。4种测定方法均可对柳树花粉进行活力检测,蓝墨水染色法可代替离体培养法对部分柳树个体的花粉活力进行快速测定。该研究揭示了柳树不同种个体间花粉变异特点,为柳树杂交育种工作提供了技术支持。

The fresh pollens were collected to optimize the in vitro germination condition from six willow species of differentparents and hybrid progeny individuals which were used as experimental materials. And four methods including in vitro culture method, blue ink staining method, I2-KI staining method and TTC staining method, were compared to explore the rapid evaluation system of pollen viability suitable for different willow individuals. The results showed that the concentration ofsucrose, H3BO3 and Ca（NO3）2 in media had slightly different effects among individuals. The culture time had significanteffects on pollen viability, with the suitable time of 10-12 h. However, the light condition had not significant effect on theculture and the optimum culture temperature was 25 °C. All of the methods for pollen viability evaluation were effective andthe blue ink staining method as a fast examination method could take place of in vitro culture method for some willow individuals. This study revealed the characteristics of pollen variation among different willow species, and provided technicalsupport for the willow hybrid breeding.