大花三色堇花粉生活力测定及贮藏方法

Pansy Pollen Viability Determination and Its Storage Method

为解决大花三色堇杂交育种过程中的时空障碍,提高育种效率,以纯黄大花三色堇品种EYO花粉为试材,采用离体萌发法、碘-碘化钾染色法、醋酸洋红染色法和TTC(氯化三苯基四氮唑)染色法对新鲜花粉生活力进行测定,并研究不同贮藏条件对花粉萌发的影响。结果表明:培养基中添加蔗糖和硼酸对花粉的萌发有促进作用,且二者之间具有互作效应,当培养基中添加200g/L蔗糖+150mg/L硼酸,萌发率较高,达35.04%。与离体萌发法相比较,碘-碘化钾染色法、醋酸洋红染色法测定的花粉活力值偏高,分别为50.80%、71.41%,TTC染色法不能使大花三色堇花粉着色。花粉在不同贮藏条件下,生活力变化存在显著差异,常温贮藏3d后花粉活力降为零,而4℃、-80℃不干燥贮藏26d后花粉仍有活力,萌发率分别为6.26%和10.31%。

In order to solve the spatio temporal obstacle in the cross breeding process, and improve the breeding efficiency of pansy, the fresh pollen ability was determined in vitro, I2 KI staining, carmine acetate dyeing and TTC staining methods, and effects of different storage on the pollen germination were studied too. The results showed that culture media with different concentration of sucrose and boric had good effects on the pollen germination. The pollen germinating percentage reached the highest after incubated on the media supplemented with 200 g/L sucrose and 150 mg/L Boric acid, the germination of pollen reached 35.04%. The rates of pollen germination were 50.80% and 71.41% detected with the methods of staining by I2 KI solution and aceto carmine respectively, these measured values were higher than the rates of germination in vitro. The fresh pollen of pansy was not colored by TTC solution. Different storage methods had significantly effected on pollen viability. The germination rate of pollen reached zero at the normal temperature for 3 days. The fresh pollen still had viability when it was stored at 4 ℃ and -80 ℃for 26 d, and the germination rates were 6.26% and 10.31% respectively.