摘要
用1%半纤维素酶,0.4%纤维素酶,0.1%果胶离析酶,CPW9M酶液分离沙打旺无菌苗下胚轴和子叶原生质体。K8P原生质体培养基悬滴培养。下胚轴原生质体形成小细胞团后用琼脂糖包埋培养,形成小块愈伤组织后转入增殖培养基M1、M2(改良MS培养基)上形成大块愈伤组织。经过两次诱导分化,在分化培养基M3(MS+0.7mg/L BA+0.2mg/L NAA),M4(MS+0.5mg/L BA+0.5mg/L KT+0.5mg/L ZT+0.2mg/L NAA)和M6(MS+3mg/L ZT+0.2mg/L IAA)上分化出苗,再生植株。由子叶分离的原生质体未能形成愈伤组织。
Protoplasts were enzymatically isolated from hypocotyls of Astragalus huangheensis. The protoplasts formed calli in K8P liquid medium. The calli were transferred onto Ml medium (MS+l-2mg/L 2,4-D + 0.5mg/L ZT + 0.2mg/L NAA) and M2 medium (MS + 0.5-1mg/L 2.4-D + 0.7mg/L BA + 0.2mg/L NAA) to form big calli. Shoot formation was initiated on M3 medium (MS + 0.7mg/L BA + 0.2mg/L NAA), M4 medium (MS + 0.5mg/L ZT + 0.5mg/L BA + 0.5mg/L KT + 0.2mg/L NAA) and M6 medium (MS + 3mg/L ZT + 0.2mg/L NAA). Plants were regenerated from protoplasts of hypocotyls.
关键词
沙打旺
原生质体培养
再生植株
Astragalus huanghecnsis
Protoplast culture
Plant regeneration
