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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。
OBJECTIVE: To optimize screening, sterilization conditions and culture medium of trichosanthes explants to reduce the pollution rate and browning rate, and to establish the trichoderma rhizome sterile culture system. Methods: The most suitable explants were screened from 5 materials of Trillium, leaves, stems, underground rhizomes and root tips. The effects of different antibacterial agents and antibiotics on the culture medium were studied. The effects of ascorbic acid (V_C), polyvinylpyrrolidone (PVP) and activated carbon on reducing browning rate were studied. Results: The rhizomes were the most suitable explants for tissue culture of Trillium chinense. Alternaria could be sterilized alternately by UV irradiation for 30 min, 800 mg carbendazim for 30 min and 0.2% Hg Cl 2 for 20 min. The medium was supplemented with 100 mg L -1 (-1) ampicillin and 1.0 g · L -1 activated carbon could reduce the tissue culture rate and browning rate of rhizome. CONCLUSION: Tricholoma glabra rhizome sterile culture system with low pollution rate and browning rate was established, which provided reference for the tissue culture of other rhizomes.