美味猕猴桃原生质体培养及植株再生技术研究

STUDIES ON TECHNOLOGY OF PROTOPLAST CULTURE AND PLANT REGENERATION OF Actinidia deliciosa

以美味猕猴桃雄株茎段愈伤组织为材料，分离原生质体，并培养在ＫＭ８Ｐ附加０．４５ｍｏｌ／Ｌ葡萄糖和０．０５ｍｏｌ／Ｌ蔗糖的培养基上，用低熔点琼脂糖包埋，６～７ｄ发生第一次细胞分裂，培养２０ｄ的分裂频率为１１．３％．在未添加新鲜培养液的情况下，原生质体再生的细胞可持续分裂至８０ｄ左右，并形成２～３ｍｍ大小的愈伤组织．然后采用二步诱导分化法将原生质体来源的愈伤组织诱导分化出绿苗，再诱导生根，形成完整的小植株．

Protoplasts isolated from stem segmentderived calli of Actinidia deliciosa (male cultivar) are cultured in KM8P medium supplemented with 0.45 mol/L-1 glucose and 0.05 mol/L-1 sucrose. Protoplasts are embeded in low melting temperature agarose. First division of regenerated cells occurrs after 6~7 days in culture and the percentage of cell division is 11.3% in 20 days. Protoplastsderived cells divide itself sustainably and develop into calli of 2~3 mm in size in 80 days in the original protoplastculturemedium without adding fresh medium. These calli regenerate shoots by using two steps differentiation method.Regenerated shoots are rooted and the perfect plantlets are regenerated.