同源四倍体黑皮冬瓜的诱导与倍性鉴定

Induction and ploidy identification of autotetraploid black wax gourd

【目的】探索高效的同源四倍体黑皮冬瓜Benincasa hispida诱变方法组合和寻求简便、快速、大通量、准确的倍性鉴定方法。【方法】以二倍体自交系幼苗为材料,利用不同浓度的秋水仙素和Oryzalin在06:00—07:00、12:00—13:00、17:00—18:00共3个时间段"剥滴"处理幼苗生长点来诱变产生同源四倍体黑皮冬瓜,利用植株形态学、细胞学及流式细胞术鉴定法与根尖染色体计数鉴定法对变异株进行倍性鉴定。【结果】不同浓度的秋水仙素和Oryzalin在不同时间段处理幼苗,均有一定的诱变效果,但秋水仙素的诱变率整体高于Oryzalin处理,且以2 g·L^(-1)的秋水仙素在06:00—07:00处理的诱变率最高,达32.18%;集成植株形态学、细胞学及流式细胞术鉴定的结果与根尖染色体计数法倍性鉴定的结果一致。【结论】2 g·L^(-1)的秋水仙素在06:00—07:00"剥滴"处理为最高效的诱变组合方法;集成植株形态学、细胞学及流式细胞术鉴定方法能简便、快速、大通量、准确地进行倍性鉴定。

【Objective】To explore efficient induction method of autotetraploid Benincasa hispida and seek a simple, convenient, large flux and accurate method of ploidy level identification.【Method】The seedlings of diploid B. hispida inbred lines were used as materials. Autotetraploid B. hispida was induced using different concentrations of colchicine and oryzalin at different times including 06：00 to 07：00, 12：00 to 13：00 and 17：00 to 18：00 to treat the growth points of seedlings. Ploidy levels of mutated plants were determined by using the methods based on plant morphology, cytology, flow cytometric analysis and chromosome numbers of root tips.【Result】There were certain induction effects by using different concentrations of colchicine and oryzalin at different times, but the induction rate of colchicine was overall higher than that of oryzalin, and the highest induction rate was 32.18% using 2 g·L（-1） colchicine from 06：00 to 07：00. The ploidy identification result fromthe combination of methods based on plant morphology, cytology and flow cytometric analysis was the same as the result based on chromosome numbers of root tips.【Conclusion】The optimum method of obtaining autotetraploid was using 2 g·L（-1） colchicine from 06：00 to 07：00. The combination of plant morphology, cytology and flow cytometric analysis was a simple, convenient, large flux and accurate way to identify ploidy level.