多花兰种子无菌萌发及离体快繁研究

以多花兰种子为材料,研究了无机盐浓度、植物生长调节剂和光照条件对多花兰种子非共生萌发的影响,在此基础上,通过研究原球茎增殖和分化、芽苗壮苗和生根的培养基配方及培养条件,建立多花兰组培快繁技术体系.结果表明:多花兰种子萌发培养基为1/6 MS十NAA0.5 mg·L-1+6-BA 2.0 mg· L-1+马铃薯泥50g·L-1+AC 1.0g·L-1,光照度为1.25μmol·m-2·s-1,萌发率63.6％；原球茎增殖继代培养基为1/4 MS+6-BA2.0 mg·L-1+NAA 0.5 mg· L1+AC 1 g·L-1+PE 200 g·L-1,繁殖倍数6.5倍/60 d,芽分化率60.2％；再生芽分化培养基为1/4 MS十6-BA2.0mg·L-1+NAA 0.2 mg· L-1+AC 1 g·L-1+PE 200g· L-1,繁殖倍数4.0倍/60 d,芽分化率85.0％；芽苗壮苗和生根培养基为1/6 MS+6-BA 3.0 mg·L-1+NAA 1.0 mg·L-1+AC 1 g·L-1+蔗糖20g·L-1 +PE 200 g·L-1和1/4 MS+6-BA 2.0 mg·L-1+NAA1.2 mg·L-1+AC1 g·L-1十蔗糖20g·L-1+PE200g· L-1,生根率达100％,生根苗移栽成活率90％.此技术可用于多花兰种苗繁育和种质资源保护.

Germfree germination of seeds and rapid prolife- ration in vitro of Cymbidium floribundum

Cymbidium floribundum seeds were used to investigate the effects of salt contents,plant growth regulators and light on seed asymbiotic germination.Based on this,the tissue culture and rapid propagation technical system of C.floribundum were established,by investigating the medium formula and culture conditions of proliferation and redifferentiation of protocorm,strong seedlings and rooting of bud.The optimal medium for asymbiotic germination of C.floribundum seeds was 1/6 MS + NAA 0.5mg·L-1+6-BA2.0mg·L-1+meshed potato 50g·L-1+AC1.0g·L-1;the light-intensity of culture was 1.25μmol·m-2·s-1;the seed germination rate was 63.6%.The optimal medium for subculture for protocorm multiplication was 1/4MS+6-BA2.0mg·L-1+NAA0.5mg·L-1+AC1.0g·L-1+PE200g·L-1;the proliferation coefficient was 6.5/60d;the bud redifferentiation rate was 60.2%.The optimal medium for bud redifferentiation was 1/4MS+6BA2.0mg·L-1+NAA0.2mg·L-1+AC1.0g·L-1+PE200g·L-1;the proliferation coefficient was 4.0/60d;the bud redifferentiation rate was 85.0%.The optimal medium for strong seedlings and rooting were 1/6MS+6-BA3.0mg·L-1+NAA1.0mg·L-1+AC1.0 g·L-1+cane sugar 20g·L-1+PE200g·L-1 and 1/4MS+6-BA2.0mg·L-1+NAA1.2mg·L-1+AC1.0g·L-1+cane sugar20g·L-1+PE200g·L-1,respectively.The rooting percentage was100%,and transplantsurvival rate reached 90%.This technology could be used to seedling breeding and germ-plasm protection of C.floribundum.