款冬叶柄愈伤组织培养与再生体系建立

以款冬幼嫩叶柄为材料,研究植物生长调节剂对其离休培养与植株再生的影响,建立了款冬离体快繁技术体系。结果表明:款冬叶柄切段较理想的灭菌方法为75%乙醇浸泡30 s,转入饱和漂白粉上清液浸泡15 min;适合愈伤诱导的培养基为MS+6-BA 3.0 mg·L~(-1)+2,4-D 2.0 mg·L~(-1),诱导率96.2%;在培养基MS+ZT 2.0 mg·L~(-1)+NAA 0.3 mg·L~(-1)上芽苗分化效果较好,分化率91%,平均芽数8.26个;较佳的不定芽增殖培养基为MS+KT 1.0 mg·L~(-1)+IBA 0.3 mg·L~(-1),增殖倍数为11.81,平均苗高4.9 cm;生根培养基为1/2MS+IBA 0.2 mg·L~(-1),生根数平均为5.68条,生根率为95.22%以上;瓶苗移栽于河沙-有机肥3∶1的基质中生长良好,成活率达90%以上;大田试验表明:同等栽培条件下款冬组培株、栽培株和野生株在生长量与花粒产量等方面存在显著差异,以组培株生长量与花粒产量相对较高。组培品与野生品有效成分含量基本一致。

Tissue culture of callus and establishment of regeneration system of Tussilago farfara petiole

Young petiole of Tussilago farfara was used as the material to investigate the plant growth regulators which could influence in vitro culture and plant regeneration and to establish rapid propagation technique. The ideal sterilization method was that young petiole of T. farfara was sterilized with 75% ethanol for 30 s, and then transferred to saturated bleaching power supernatant for 15 min. The suitable medium for callus induction was MS+6-BA 3.0 mg·L^-1 +2,4-D 2.0 mg·L^-1 with 96.2% induction rate. The seedlings had better differentiation with 91% differentiation rate and 8.26 buds on the medium containing MS+ZT 2.0 mg·L^-1 +NAA 0.3 mg·L^-1. The preferred enrichment medium of adventitious bud was MS+KT 1.0 mg·L^-1 +IBA 0.3 mg·L^-1 with 11.81 enrichment times and 4.9 cm seedling height. The rooting medium included 1/2MS+IBA 0.2 mg·L^-1 with the average number of rooting was 5.86 and the rooting rate was above 95.22%. The container seedlings can grow well and the survival rate was more than 90% when they were transplanted on the medium added with river sand and organic fertilizer with the ratio of 3:1. The field experiments indicated that significant differences in increment and yield of pollen grains among the tissue-culture, cultivation and wild type of T. farfara under the same cultivation conditions. The cultivated plants were relatively high on the increment and yield of pollen grains.The active ingredient contnet of the tissue culture and the wild materials was basically the same.