矮牵牛游离小孢子培养体系的建立及关键影响因素分析OA

Taking four Petunia hybrida genotypes,namely'Mitchell''Rose Star Spot New Superb''Mirage Exquisite Pink'and'Dreamy Burgundy'as experimental materials,the method of isolated microspore culture was employed to investigate the impacts of factors including the correspondence between the external morphology of flower buds and the development stages of microspores,disinfection methods,genotype differences,low-temperature pretreatment,combined pretreatment of starvation and high-temperature,and hormone treatment on the effects of isolated microspore culture.The objective was to establish an isolated microspore culture system for Petunia hybrida,in order to provide a reference for the haploid breeding research of Petunia hybrida and other Solanaceae ornamental plants.The results showed that the sampling periods for the four varieties corresponded to petal-sepal ratios of 0.48,0.73,0.60,and 0.58 respectively.There were significant differences in induction effects among different genotypes,with'Mitchell'having the highest induction rate.Disinfection with 1％NaClO for 25 minutes could reduce the contamination rate of microspores and maintain their high activity.Low-temperature pretreatment(treatment at 4 ℃ for 6 days)and combined pretreatment of starvation and high-temperature(3 days high-temperature treatment at 32 ℃+6 days starvation treatment)could significantly increase the induction rate of embryoids.Hormone combinations of TDZ at 0.5 mg·L-1 and 2,4-D at 0.5 mg·L-1+6-BA at 0.2 mg·L-1 could promote the formation and development of embryoids.