6个铁线莲品种花粉保存方法
Pollen preservation methods for six Clematis florida cultivars

为了探究铁线莲花粉保存的最佳条件,研究比较了花粉采集时间(开花第1、2、3天)、干燥时间(12、24、48 h)和保存温度(4、-20、-196 ℃)对花粉萌发率的影响。结果表明,花粉粒的平均直径均随着开花时间的延长而上升,且差异显著,其中‘蓝龙’花粉粒直径上升幅度最大为4.53 μm,‘如梦’的上升幅度最小,为1.13 μm。开花第2天采集的‘爱莎’、‘第一夫人’、‘冰美人’、‘亨利’和‘如梦’花粉萌发率最高,而开花第3天采集的‘蓝龙’花粉萌发率最高;‘第一夫人’、‘爱莎’和‘蓝龙’分别经过12~48 h干燥后4 ℃保存的花粉相对存活率最高,分别为新鲜花粉的87.86%、71.27%和67.24%;‘亨利’、‘冰美人’和‘如梦’花粉干燥24 h后-196 ℃保存的相对存活率最高,分别为新鲜花粉的59.65%、78.74%和69.33%。-20 ℃保存的铁线莲花粉相对存活率仅为0~56.85%。利用4和-196 ℃保存的花粉进行杂交验证试验,平均坐果率为70.7％~100%,初步明确了‘第一夫人’、‘爱莎’和‘蓝龙’花粉适合4 ℃保存,而‘亨利’、‘冰美人’和‘如梦’则适合液氮超低温保存,这为解决铁线莲品种间杂交的花期不遇问题奠定了基础。
In order to investigate the optimum preserving conditions for pollens of Clematis florida,the effects of collecting time (the first,second and third day of flowering),drying time (12,4 and 48 h) and preserving temperature (4,-20 and -196 ℃) on the pollen germination rates were tested.The results showed that the average diameter of pollen grains was increased along with the flowering time.The differences among six cultivars were significant,which the maximum amplitude was 4.53 μm for C.‘Fujinami’ and with a minimum of 1.13 μm for C.‘Hagley Hybrid’.The pollen germination rates reached the peak at the second day of flowering for C.‘Asao’,C.‘The First Lady’, C.‘Madame le Coultre’,C.‘Henryi’ and C.‘Hagley Hybrid’,but at the third day of flowing for C.‘Fujinami’.The pollens of C.‘The First Lady’,C.‘Asao’ and C.‘Fujinami’ stored at 4 ℃ obtained the highest relative survival rate of 87.86%,71.27% and 67.24% after drying for 12-48 h.After preserved at -196 ℃,the relative survival rates of the pollens of C.‘Madame le Coultre’,C.‘Henryi’ and C.‘Hagley Hybrid’after drying for 24 h were 59.65%,78.74% and 69.33%,respectively,only 0~56.85% of the pollens of the six cultivars could be active preserved at -20 ℃.The averaged fruit setting rates of crossbreed were 70.7%~100% when pollens preserved at 4 ℃ or -196 ℃.The above results showed that the pollens of C.‘Asao’,C.‘Fujinami’ and C.‘The First Lady’ should be preserved at 4 ℃,while the ones of C.‘Madame le Coultre’,C.‘Henryi’ and C.‘Hagley Hybrid’ should be preserved at -196 ℃.The present study laid the foundation for solving the flowering asynchronism in crossbreeding.