延龄草无菌培养体系的建立及防褐变措施

目的:对延龄草外植体的筛选及灭菌条件和培养基进行优化,降低污染率和褐变率,建立延龄草根茎无菌组织培养体系。方法:从延龄草子房、叶、茎、地下根茎、根尖5种材料中筛选出最适宜的外植体,考察多种药液交替灭菌和培养基中添加抗生素对降低组培污染率的影响,研究培养基中添加抗坏血酸(V_C),聚乙烯吡咯烷酮(PVP),活性炭对降低褐变率的效果。结果:地下根茎为延龄草组织培养最适宜的外植体,可采用紫外线照射30 min,800倍多菌灵浸泡30 min和0.2%Hg Cl_2处理20 min交替灭菌,培养基中添加100 mg·L~(-1)氨苄青霉素和1.0 g·L~(-1)活性炭可降低根茎组培污染率和褐变率。结论:建立了低污染率和褐变率的延龄草根茎无菌培养体系,为其他根茎类植物组织培养提供参考。

Establishment of Sterile Culture System of Trillium tschonoskii and Control of Browning

Objective: To optimize the screening, sterilization and culture medium for explants of Trillium tschonoskii, reduce the pollution rate and browning rate, and establish an aseptic tissue culture system of T. tschonoskii. Method: Most suitable explants were selected from ovary, leaves, stems, underground rhizomes and root tips of T. tschonoskii, and the effect of a variety of drug liquid alternating sterilization and addition of antibiotics in medium were studied on the contamination rate. VitC(V_C), polyvinyl pyrrolidone(PVP) and activated carbon were added to the medium to study their effect on reducing the browning rate. Result: The underground rhizome of T. tschonoskii was the most optimum explants. It could be alternately sterilized with ultraviolet radiation for 30 min, 800 times of carbendazim soaking for 30 min, and processed with 0.2%HgCl₂ for 20 min. The medium supplemented with 100 mg · L^-1 ampicillin and 1.0 g · L^-1activated carbon could reduce the contamination rate and browning rate of the rhizome tissue culture. Conclusion: The rhizome sterile culture system of T. tschonoskii has been established with low contamination rate and browning rate. It can provide reference for tissue culture of other rhizome-plants.