金花茶离体培养体系的建立

目的]通过愈伤组织发生途径建立金花茶(Camellia nitidissima)的离体培养体系,为金花茶的快速繁殖和种质资源保存提供理论和技术支持.方法]在金花茶种子萌发阶段以沙子替代培养基进行无菌萌发,以其成熟植株的茎尖、茎段和叶片为外植体,在WPM培养基上添加不同种类和浓度激素进行愈伤组织诱导、愈伤组织分化及生根培养.结果]金花茶成熟种子去壳、去皮、取出胚乳后在添加无菌液的湿润沙子中培养,平均萌发率为95.40%,平均萌发时间为19.10 d;WPM+2.00 mg/L 6-BA+0.50 mg/L 2,4-D是金花茶子叶、胚轴愈伤组织诱导的适宜培养基;WPM+2.00 mg/L 6-BA+0.10 mg/L NAA是愈伤组织分化的适宜培养基,分化系数为4.56;1/2WPM+0.50~1.00 mg/L IBA是金花茶生根的适宜培养基,生根率为45.40%~54.00%,生根时间为19.80~20.90 d.结论]金花茶离体培养可通过胚轴诱导产生愈伤组织进而分化成完整植株.

Establishing in vitro culture system of Camellia nitidissima

Objective]The present study established in vitro culture system of Camellia nitidissima through callus ap-proach to provide theoretical and practical support for rapid reproduction and germplasm resources preservation. Method]At seed germination stage, sand was used instead of medium for aseptic germination. Shoot tip, stem and leaf of mature plant were used as explants. Callus induction,callus differentiation and rooting culture were conducted in WPM media sup-plemented with various hormones at different densities. Result]The mature seeds were peeled, and their endosperms were removed. Endosperm-removed embryos were buried in the sand wetted by sterile liquid. In such condition, average germi-nation rate was 95.40%, average germination time was 19.10 d. The suitable culture medium for callus induction of cotyle-don and hypocotyl was WPM+2.00 mg/L 6-BA+0.50 mg/L 2,4-D. The suitable culture medium for callus differentiation was WPM+2.00 mg/L 6-BA+0.10 mg/L NAA, and differentiation coefficient was 4.56. The optimum culture medium for rooting culture was 1/2WPM+0.50-1.00 mg/L IBA,and rooting rate was 45.40%-54.00%,rooting time was 19.80-20.90 d. Conclu-sion]The in vitro culture system of C. nitidissima is as follows:callus is induced by hypocotyl, then callus differentiate into whole plant.