摘要
用酶法由甘蓝型油菜(Brassica napus L.)下胚轴分离出原生质体,在Nitsch培养基(添加1 mg·L^(-1)2,4-D、1 mg·L^(-1)NAA、O.5mg·L^(-1)BAP及0.49mol·L^(-1)蔗糖)中进行液体浅层培养。再生的细胞团植板于MS固体培养基上,培养基中添加1mg·L^(-1)BAP、0.1mg·L^(-1)2,4-D(或2 mg·L^(-1)BAP、 0.1mg·L^(-1)NAA),3%蔗糖及0.5%琼脂糖。待长成直径约5mm的小愈伤组织后转到MS固体分化培养基(添加2mg·L^(-1)BPA及1%琼脂)上,再生了植株。并对影响培养的若干因素进行了探讨。
Protoplasts isolated from hypocotyls of Brassica napus were inoculated in Nitsch medium supplemented with 1 mg·L^(-1)2, 4-D, 1 mg·L^(-1)NAA, 0.5mg· L^(-1)BAP and 0.49 mol·L^(-1)sucrose at a density of 10~4～10~5 protoplasts/ml and maintained at 25℃ in darkness. Fresh medium was added at an interval of every three days since the second week. The protoplasts could undergo first division in 1～2 days and second division in 5～6 days after inoculation. Large multicellular masses and microealli which were visible after 15～20 days of cuture were transferred onto MS medium supplemented with 0.1 mg·L^(-1)2, 4- D, 1 mg·L^(-1)BAP, 3% sucrose and solidified with 0.5% agarose, and kept at 25℃ with 16 hrs illumination per day. Approximately 15 days later, the calli that grew to ca. 5 mm in diameter were then transferred onto MS medium supplemented with 2 mg·L^(-1)BAP, 3% sucrose and 1% agar for diffcreentiat- ion. 12 plantlets were obtained, among them one plantlet has flowered in the flask.
出处
《武汉大学学报（自然科学版）》
CSCD
1992年第3期97-101,共5页
Journal of Wuhan University(Natural Science Edition)
基金
国家自然科学基金
高等学校博士点研究基金资助的课题
关键词
甘蓝型
油菜
原生质体
再生
植株
protoplast culture
plant regeneration
Brassica napus
