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以南瓜(Cucurbita moschata)未授粉子房为外植体,在离体条件下研究了激素种类、基因型、胚囊发育时期、消毒方式和热激处理时间对胚状体诱导效果的影响,旨在建立完善的南瓜未授粉子房离体诱导再生体系。结果表明,MS+4.0mg·L–1 2,4-D+0.5 mg·L–1 NAA+1.0 mg·L–1 6-BA和MS+0.04 mg·L–1 TDZ两种培养基胚状体的诱导率都较高,分别为19.8%和20.1%,二者相比,使用TDZ诱导更加简单;在6个南瓜品种中,2个生长势较强的品种(雪峰蜜本和鱷鱼南瓜)胚状体诱导率较高,适合作为离体雌核诱导的实验材料;开花当天的子房先切片后消毒可以有效降低愈伤组织的形成,提高胚状体的诱导率;在黑暗且35°C条件下热激5天有利于子房的转绿和出胚。将子叶胚转移至无激素的MS培养基上得到再生植株并移栽成活。经鉴定有7个再生植株的染色体数目n=x=22,气孔保卫细胞叶绿体数的平均值为4.28个,是单倍体植株。
The effect of hormone type, genotype, embryo sac development period, disinfection method and heat shock treatment time on the induction of embryoid bodies was studied in Cucurbita moschata without pollination ovary. In the establishment of a sound pumpkin unpollinated ovary in vitro induced regeneration system. The results showed that the embryos of MS + 4.0 mg · L-1 2,4-D + 0.5 mg · L-1 NAA + 1.0 mg · L-1 6-BA and MS + 0.04 mg · L- The results showed that induction of TDZ was more efficient than that of TDZ induction. Compared with the control, TDZ induction was more simple. Among 6 pumpkin cultivars, 2 plants with stronger growth potential (Xuefeng honey and crocodile pumpkin ) Embryoid body higher induction rate, suitable as an in vitro gynogenetic experimental material; on the day of flowering ovary first section after disinfection can effectively reduce the formation of callus and improve embryoid body induction rate; in the dark and 35 Heat treatment at ° C for 5 days favored greening and embryogenesis in the ovaries. Cotyledonary embryos were transferred to hormone-free MS medium to obtain regenerated plants and transplanted to survive. The number of chromosomes of 7 regenerated plants was identified as n = x = 22, and the average number of stomatal guard cells was 4.28, which was haploid plants.