虎杖茎尖离体再生体系的建立和优化

ESTABLISHMENT OF OPTIMAL REGENERATION SYSTEM in vitro FOR STEM APEXES OF Polygonum cuspidatum

研究了苗龄、接种方式、不同植物生长调节剂、硝酸银和蔗糖等对虎杖(Polygonum cuspidatum)茎尖愈伤组织诱导、不定芽分化和生根的影响。结果表明:愈伤组织诱导的最佳培养基是MS+蔗糖28g/L+琼脂5.5g/L+2,4-D 1.5mg/L+6-BA 1.0mg/L,诱导率为100%;3-7d苗龄的虎杖茎尖愈伤组织诱导能力无显著差异,诱导率均达到95%以上,此后随着苗龄的增加,愈伤组织诱导能力快速下降,苗龄为12d时愈伤组织诱导率只有55.6%;以正插(形态学下端插入培养基)方式接种的外植体愈伤组织诱导率显著大于反插(形态学上端插入培养基)和平放的;不定芽分化最佳培养基为MS+AgNO34.0mg/L+蔗糖30 g/L+琼脂6.0g/L+NAA 0.5mg/L+TDZ 0.8mg/L,分化率为83.9%,增殖系数为7.63;生根培养基选用1/2 MS+蔗糖26g/L+琼脂6.5g/L+活性碳3%+IBA 0.2mg/L+NAA 0.3mg/L,生根率为100%。用透气膜封口比用聚乙烯菌膜生根率明显提高,生根明显提前。

The regeneration system of Polygonum cuspidatum in vitro was studied with stem apexes as explant. The effects of seedling age, inoculation method, different plant growth regulators, AgNO3 and sucrose on callus induction, bud differentiation and rooting were investigated. The results showed that the optimal medium for callus induction was MS ＋ sucrose 28g/L ＋ agar5.5g/L ＋ 2,4-D 1.5mg/L ＋ 6-BA 1.0mg/L with induction frequency of 100%. There was no significant difference among 3 to 7d seedling in their callus induction frequency, which were above 95 %. And there after the increasing of seedling age resulted in a reduction of callus induction frequency, which was only 55.6 % at the seedling age of 12d. The stem apexes inoculated with their abaxial end up were more effective in callus induction frequency than their abaxial ends down or placed horizontally. Best bud differentiation and bud multiplying medium with bud differentiation rate of 83.9% and multiplying coefficient of 7.63 was ： MS ＋ AgNO3 4. 0mg/L ＋ sucrose 30g/L ＋ agar 6.0g/L ＋ NAA 0.5mg/L ＋ TDZ 0. 8mg/L. As for the rooting induction, the best medium with the rooting rate of 100% was： 1/2 MS ＋ sucrose 26g/L ＋ agar 6.5g/L ＋ activated carbon 3% ＋ IBA 0.2mg/L ＋ NAA 0.3mg/L. In root induction culture using membrane filter the rate of root formation was higher and earlier than that of root formation with common membrane.