木本曼陀罗毛状根植株再生体系的建立

Plant Regeneration from Hairy Roots of Datura arborea L.

摘要:

建立了木本曼陀罗(Datura arborea L.)毛状根的植株再生体系并对再生植株进行了初步检测。采用“一步法”诱导不定芽的适宜培养基为MS+6-BA 2.0 mg L-1+NAA 0.2 mg L-1；采用“两步法”诱导不定芽时, 先在MS+6-BA 4.0 mg L-1 +KT 0.5 mg L-1+2,4-D 0.5 mg L-1上诱导愈伤组织, 然后在MS+6-BA 4.0 mg L-1+NAA 0.2 mg L-1上诱导愈伤组织分化不定芽。诱导的不定芽的最适宜生根培养基为1/2MS+IBA 0.1 mg L-1。用PCR技术从再生植株叶片中得到了rolB、rolC目的基因片段。HPLC检测结果表明毛状根再生植株中莨菪烷类生物碱(Tropane alkaloids, TA)的含量较野生植株有明显的提高。

关键词:木本曼陀罗;毛状根;再生;莨菪碱;东莨菪碱

Abstract:

Plant regeneration system derived from hairy roots of Datura arborea L. was established. The optimum medium for inducing adventitious buds by ‘one-step method’ was MS+6-BA 2.0 mg L-1+NAA 0.2 mg L-1. Using ‘two-step method’, callus was induced on MS medium supplement with 6-BA 4.0 mg L-1+KT 0.5 mg L-1+2,4-D 0.5 mg L-1 at first, then adventitious buds were differentiated from the induced callus on MS medium supplemented with 6-BA 4.0 mg L-1+NAA 0.2 mg L-1. The optimum rooting medium for adventitious buds was 1/2MS + IBA 0.1 mg L-1. By using PCR technique, the gene segments rolB and rolC were obtained from the leaves of regeneration plant. The results of HPLC showed that the contents of tropane alkaloids (TA) in regeneration plants was much more than those in the wild plants.

Key words:Datura arborea L.; Hairy roots; Regeneration; Hyoscyamine; Scopolamine