濒危植物珙桐愈伤组织的诱导及悬浮细胞培养初探

Callus Induction and Preliminary Study of Cell Suspension Culture in an Endangered Plant,Davidia involucrata

以珙桐(Davidia involucrate)无菌苗的下胚轴为外植体,以LSD分析和正交设计与分析为主要研究方法。诱导和筛选愈伤组织,进行悬浮培养,初步建立了珙桐悬浮培养体系,并对珙桐悬浮培养中生物量和pH值的变化进行了初步研究。结果表明:KT和2,4-D的组合诱导出的愈伤组织更适合进行细胞悬浮培养,最佳愈伤组织诱导培养基为WPM+KT 1.0 mg/L+2,4-D 0.5 mg/L+Vc 1.0 mg/L;悬浮细胞培养中,接种量和Ca2+浓度是影响生物量的主要因素,最佳接种量为25 g/L,最佳Ca2+浓度为标准WPM培养基中Ca2+浓度的2倍;黑暗条件下,珙桐悬浮培养生物量变化曲线呈"S"型,在21 d时可获得最大生物量;pH值的变化与生物量的变化有一定的相关性。

The hypocotyls from aseptic seeding of Davidia involucrata were studied via LSD analysis and orthogonal design.We firstly induced and screened the callus and then carried out suspension cultures.On this basis,we established the cell suspension culture system of Davidia involucrata,and determined the changes in biomass and pH.The results showed that the hormone combination of KT and 2,4-D induced callus were more suitable for cell suspension culture,and the best callus induction medium was WPM with KT 1.0 mg/L,2,4-D 0.5 mg/L and Vc 1.0 mg/L.The quantity of the original incubated callus and the concentration of Ca2＋ were the main factors affecting biomass in the cell suspension culture system.The best quantity of original incubated callus was 25 g/L,and the best concentration of Ca2＋ was two times the concentration of Ca2＋ in the standard WPM medium.Under dark conditions,the growth change curve for the biomass suspension culture of Davidia involucrata resembled an＂S＂and the maximum biomass was available in 21 d.Results also showed a relationship between the change in pH and changes in biomass.