甘蔗原生质体培养──快速再生植株

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用MSCa培养基从甘蔗PC9102嫩叶诱导愈伤组织。接种30~40天后,将愈伤组织转入AA液体培养基,进行悬浮培养、经过50~80天,建立起典型胚性细胞悬浮培养物,分离到原生质体、原生质体用MRP1-MRP2琼脂糖培养基培养,16~20天后给予光照,25~30天后转入Ne分化培养基,45天左右可见绿芽分化。重复试验表明,从外植体诱导愈伤组织开始,建立悬浮细胞系,分离原生质体到培养再生完整小植株的全过程约200天。 Callus was induced from young leaves of sugarcane PC9102 using MSCa medium. After 30 to 40 days of inoculation, the callus was transferred to AA liquid medium and cultured in suspension. After 50 to 80 days, a typical embryogenic cell suspension culture was established and protoplasts were isolated. Protoplasts were stained with MRP1-MRP2 agarose After 16 to 20 days light irradiation was given, and 25 to 30 days later it was transferred to Ne differentiation medium. About 45 days, the differentiation of green shoots was observed. Repeated experiments showed that starting from the explant-induced callus, the establishment of a suspension cell line, protoplast isolation and regeneration of whole plants to regenerate the entire process of about 200 days.
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