灌木铁线莲多倍体诱导与鉴定

Polyploid induction and identification of Clematis fruticosa

采用秋水仙素棉球处理法和琼脂处理法对灌木铁线莲进行诱变处理,以探究秋水仙素诱导灌木铁线莲多倍体的最佳条件和鉴定方法,成功获得了同源四倍体新种质(2n=4x=32)。研究结果表明,对于灌木铁线莲的秋水仙素诱变处理,棉球处理法要优于琼脂处理法。棉球处理法以0.2%~0.3%浓度的秋水仙素下处理48~72 h的诱变效果较好,琼脂处理法在0.2%浓度下处理96 h的变异率最高,为20%。琼脂处理可通过延长处理时间增强诱变效果。秋水仙素处理过的植株生长发育较为缓慢、株高降低,叶面积、叶片气孔变大。在对嵌合体不同倍性细胞含量的分析中,采用图像分析技术对气孔鉴定法进行创新,获得的嵌合率与流式细胞仪的鉴定结果呈极强相关性,推测此方式在多倍体鉴定中具有应用价值。

In order to explore the optimum conditions and identification methods of colchicine-induced polyploidy of Clematis fruticose,colchicine cotton ball treatment and agar treatment were used to mutagenize C.fruticosa.A new tetraploid germplasm(2n=4x=32)was successfully obtained.The results showed that the cotton ball treatment was superior to the agar treatment for colchicine mutagenesis of C.fruticosa.The cotton ball treatment with 0.2%-0.3%colchicine for 48-72 h had better mutagenic effect.The mutation rate of agar treatment at 0.2%concentration for 96 h was 20%,was the highest,and agar treatment could enhance mutagenic effect by prolonging treatment time.The plants treated with colchicine grew slowly,plant height decreased,leaf area increased,leaf stomata enlarged.In the analysis of different ploidy cell contents of chimeras,image analysis technology was used to innovate the stomatal identification method.There was a strong correlation between the chimerism rate obtained and the results of flow cytometry,which suggested that this method had application value in polyploid identification.