山杏原生质体培养再生植株

Plant Regeneration from Cultured Protoplasts of Armeniaca vulgaris var. ansu

对影响山杏原生质体分离和培养的因素进行了系统研究。以悬浮培养物为分离材料，在ＣＰＷ＋１．５％ＣｅｌｕｌａｓｅＯｎｚｕｋａＲ－１０＋０．５％ＭａｃｅｒｏｚｙｍｅＲ－１０＋０．５％Ｈｅｍｉｃｅｌｕｌａｓｅ＋０．６ｍｏｌ／Ｌ甘露醇＋１％ＰＶＰ的酶解液中酶解１０ｈ，原生质体产量和活力分别达到８×１０６个／ｇＦＷ和９５％。以ＫＭ８Ｐ为基本培养基，在培养初期加２，４－Ｄ１．０、ＢＡ０．２５ｍｇ／Ｌ，培养１０ｄ和３０ｄ后分别将ＢＡ调至０．５和１．０ｍｇ／Ｌ，以０．５５ｍｏｌ／Ｌ山梨醇调节渗透压，培养过程中逐步降压，在０．５×１０５个／ｍＬ的植板密度下液体浅层培养６０ｄ后形成了微愈伤组织。将微愈伤组织转至固体培养基上继代培养，在分化培养基上分化出不定芽，在生根培养基上生根形成完整植株。

The conditions of protoplast isolation and culture of Armeniaca vulgaris var. ansu were studied systematically.The protoplast yield and viability could amount to 8×10 6protoplasts/g FW and 95% respectively when suspension cultures were incubated in enzyme solution containing CPW salts,1.5% Cellulase Onzuka R-10,0.5% Macerozyme R-10,0.5% Hemicellulase, 0.6 mol/L mannitol and 1% polyvinylpirrolidone for 10 hours.Protoplasts were cultured on KM8P medium with 1.0mg/L 2,4-D and 0.25mg/L BA at beginning stage,adjusting BA concentration to 0.5 and 1.0mg/L after 10 days and 30 day′s culture respectively.Osmoticum of the medium was adjusted to 0.55mol/L with sorbitol and the osmotic pressure was reduced progressively.Microcalli formed after 60 days′s culture in a liquid layer with plating density of 0.5×10 5/mL.The microcalli were subcultured on solid medium and developed adventitious buds on differentiation medium.Eventually,shoots rooted and developed into whole plantlets on rooting medium.