铁线莲‘Utopia’无菌外植体的建立

Studies on Getting Bacteria-free Explants of ‘Utopia’ by Applying Orthogonal Design

以Clematis‘Utopia’为材料,采用L9(34)正交试验设计,设外植体(A:分别为叶片、带一对侧芽茎段和不带侧芽茎段)、消毒时间(B:消毒时间分别为3、57、min)和培养基(C:分别为培养基:MS+0.75 TDZ+0.1 NAA、MS+0.75 TDZ+1.0 2,4-D和MS+0.5 TDZ+1.0 2,4-D)3因素,每因素设3个水平。外植体经表面消毒后,接种在MS琼脂培养基中,于(25±1)℃,2 000lx光照培养,25 d后统计成活率。结果表明:最优水平组合为A1B1C2。

The Clematis ＇Utopia＇ was used as test material, test method was orthogonal design L9 （34 ）. Three factors were explants（A）, sterilizing times （B） and culture medium （C）. Explants were leaves, stem with axillary buds and stem without axillary buds. Sterilizing times were 3 rain, 5 min and 7 min. Culture medium were MS＋0. 75 TDZ＋0. 1 NAA,MS＋0, 75 TDZ＋1. 0 2,4-D and MS＋0. 5 TDZ＋1.0 2,4=D. Explants were inoculated on MS base medium（supplied with 7 g/L agar and 30 g/L sucrose, pH 5. 8） after them were sterilized, and cultured under （25±1）℃ and 2 000 lx conditions, then the explants were compiled surviving rate after cultured 25 days. The results showed that the best level combination was A1B1C2.