目的为老瓜头Cynanchum komarovii资源利用提供前期的生物技术实验参考。方法以老瓜头的根段、茎段和叶片为外植体,采用正交试验方法,筛选诱导愈伤组织产生的最适培养基;进行愈伤组织增殖培养过程中,测定了不同时间愈伤组织的鲜、干质量,绘制愈伤生长曲线;同时进行了愈伤组织分化培养基的实验筛选。结果不同外植体诱导愈伤产生,其根诱导效果最好,各处理诱导率达90%~100%,茎段和叶片愈伤组织诱导最佳配方分别为MS+2,4-D1.0mg/L+6-BA2.0mg/L+NAA0.5mg/L和MS+2,4-D0.5mg/L+6-BA0.5mg/L+NAA0.5mg/L,茎段比叶片较容易诱导,各处理诱导出的愈伤组织质地松散,颜色鲜黄,几乎无褐化现象。愈伤组织继代生长在第10天进入对数生长期,愈伤鲜生长量最大达4.961g,干质量0.496g。愈伤组织的分化率很低,很难生芽,易分化出钉状根。结论对于诱导愈伤形成,根段是最佳的外植体,筛选出了茎段和叶片诱导愈伤的最适配方,愈伤组织生长量较大,但分化困难。

Objective To provide the initiative experimental reference for resource utilization of Cynanchum komarovii i by biological technology. Methods Using root, stem, and leaf of C. komarovii as explants, the proper media for inducing of callus were optimized by orthogonal test. In culturing process for proliferation of callus, the fresh and dry weights were determined at various times and callus growth curve was drawn. Meanwhile, division test of callus was taken. Results Root callus test showed the best effect in successful inducing percentage of 90%-100%. The best media combination of stem and leaf were MS+2,4-D 1.0 mg/L+6-BA 2.0 mg/L+NAA 0.5 mg/L and MS+2,4-D 0.5 mg/L+6-BA 0.5 mg/L+NAA 0.5 mg/L. The stem segament was more easily induced than leaf. The callus induced by various treatments is in loose character and fresh yellow colour with no brown phenomenon. At day 10 the growth entered the exponent period of callus offspring, the maximal fresh weight to 4.961 g and the dry weight to 0.496 g. The division rate of callus was rather low and not easy to shoot. However, the nail root was commonly seen in the test. Conclusion The root is proved to be the best explants in the test of inducing callus, which shows the best medium of stem and leaf for forming the callus. The growth of callus is bigger, but the division is not easy.

宁夏大学自然科学基金资助项目(NS0505)