摘要： 用决明（CasiaobtusifoliaL.）子叶和下胚轴为材料，游离和培养原生质体。结果表明，15日龄无菌苗的子叶和下胚轴比较适于游离原生质体，每1g材料的原生质体产量高达1.6×104个；PectolyaseY-23是分离原生质体所必需的；用含2,4-D0.4mg/L（以下单位同）,NAA1.0与KT0.1的KM8P漂浮培养利于原生质体的分裂。培养4d后，原生质体开始分裂，15d后其植板率约为19.2%,30d形成了细胞团或小愈伤组织。增殖愈伤组织在分化培养基上培养，可分化出芽。芽在生根培养基上培养14d生根，从而再生决明小植株。
关键词:
决明,
原生质体,
再生植株
Abstract: The culture of protoplast derived from cotyledon and hypocotyl of Cassia obtusifolia seedlings was studied.About 1.5mm long sections of cotyledons and hypocotyls were digested in any enzyme solution for a certain time.The protoplasts were cultured in a liquid medium at a density of 5.1×10 4 No./ml and transferred onto a solid medium for callus culture and shoot regeneration.Some factors affecting protoplast isolation and division were studied.The results showed that 15 day age seedlings’ hypocotyls and cotyledons used as donor tissues were suitable for their protoplast isolation and the yield of their protoplasts amounted to 1.6Ⅹ10 4NO /g.FW;Pectolyase Y 23 was indispensible to their protoplast isolation;KM8P medium supplemented with 0.4mg/L 2,4 D,1.0mg/L NAA and 0.1mg/L KT as well as floating culture fitted in the division of protoplast derived cells.After a culture of 3 days ,the first division was observed.The plating frequency was about 29.2% in 15 days.Complete plantlets were regenerated 14 days after the shoots,3 cm or so,were transferred onto 1/2 MS medium containing 0.2mg/L IBA.
Key words:
Cassia obtusifolia,
Cotyledon and hypocotyl protoplast,
Plant regeneration
中图分类号:
周延清， 苑保军， 张根发， 卢龙斗. 决明原生质体的分离与培养研究[J]. 华北农学报, 1998, 13(3): 107-111. doi: 10.3321/j.issn:1000-7091.1998.03.021.
Zhou Yanqing， Yuan Baojun， Zhang Genfa， Lu Longdou. A Study of Influence Factors on Isolation Culture of Cotyledon and Hypocotyl Protoplasts from Cassia obtusifolia Seedlings[J]. ACTA AGRICULTURAE BOREALI-SINICA, 1998, 13(3): 107-111. doi: 10.3321/j.issn:1000-7091.1998.03.021.