白术不定根的高效诱导及悬浮培养研究

Studies on Adventitious Root Suspension Culture of Atractylodes macrocephala Koidz.in Vitro

对白术试管苗不定根的诱导和离体培养进行了系统研究。通过在培养基中添加不同浓度的蔗糖和生长素,筛选并建立了能够从白术叶片有效诱导不定根的最适培养基:MS+蔗糖4%+NAA(α-萘乙酸)2.0 mg/L;通过正交试验的方法对所得到的不定根进行悬浮培养研究,得到最适培养基:MS+蔗糖3%+NAA1.0 mg/L+ABT(生根粉)0.5 mg/L。对悬浮培养得到大量根在最适培养基上进行继代培养并测定得到生长曲线。同时,使用高效液相对不定根和药材进行对比分析,发现培养物中含有和作为中药材的白术根茎中相同的各类化学成分。为白术中有效成分的获得手段提供了一种新思路;证明了某些中药植物药用部位可以成功地进行单独培养,并且预示有可能通过发酵罐培养获得大量目的产物。

The induction and culturc of adventitious root of Atracylodes macrocephala Koidz. were studied in vitro systemically. To establish a new technique for direct induction adventitious roots from the leaves of Atracylodes macrocephala Koidz. , basal MS medium supplemented with different concentrations of sucrose and growth regulators was tried and optimized. The experimental results indicated that the optimized medium for efficient adventitious root induction was MS--Sucrose 4% ＋NAA2.0 mg/L, on which clustered roots formed and grew vigorously through 2 months. The optimal medium for suspension culture of roots was determined by using orthogonal design, and the results showed that the most appropriate condition for root mass accumulation was MS＋Sucrose 3%＋NAA 1.0 mg/L＋ABT 0. 5 mg/L. Growth curve on optimal suspension culture medium was also determined. Meanwhile, fingerprint spectra showed that the cul tured material contained the same chemical components as the original TCM. This research will provide techniques and new thoughts for obtaining effective chemical ingredients.