秦艽发根系体细胞胚的诱导和植株再生

Induction of somatic embryos and plant regeneration from hairy root line of Gentiana macrophylla pall

在含0.1—10.0 mg/L 2,4-D的MS培养基上,秦艽发状根切段在30d后全部转化成淡黄色致密的胚性愈伤组织.转入不含激素的MS培养基上继续培养30d后,该愈伤组织转变成由许多淡黄色致密颗粒(球形的体细胞胚)组成的疏松愈伤组织团.把愈伤组织团切成小块,再次转到新鲜的MS无激素固体培养基上培养30d后,表面有许多独立的和丛生的芽出现,解剖镜下观察发现,该愈伤组织团内包含许多球形、心形、鱼雷形、子叶形胚.将所得到的再生小植株移栽到土/蛭石混合物(1:1)中后,大多数再生苗可以成活.与野生苗相比,再生苗的叶子看起来相对加宽,叶面积增大,叶色较深.胚性愈伤组织诱导的最佳培养基为含1.0 mg/L 2,4-D的MS培养基.

30 day after having been cultured on sold MS medium with 0.1-10.0 mg/L 2, 4-D, all the hairy root segments of Centiana macrophylla Pall., produced by infecting the wounded cotyledon with Agrebaterium rhizogenes, converted into yellowish compact embryogenic calli. 30 day after having been transferred onto hormone free MS medium, these calli developed into loose yellowish calli consisting of many small compact particles. Another 30 days after being cut into smaller pieces and transferred onto fresh MS agar medium without hormone, many single and fasciculate shoots formed on the surfaces of these calli. Under the anatomical microscopy, these cultures were found to contain many globe-, heart-, torpedo- and cotyledon- shaped somatic embryos and plantlets with well-developed roots. The majority of the regenerated plantlets were able to survive after being transplanted in a mixture of soil and vermiculite（1 ： 1）. In comparison with corresponding wild seedlings, those regenerated plantlets displayed widened and color-darkened leaves. The optimal medium which could produce somatic calli is MS medium with 1.0 mg/L 2, 4-D.