Protoplastfusion or somatic hybridization technology is an alternativetechnology for production hybrids of plants that are difficultto be produced by conventional methods due to their sexualincompatibility. An experiment was conducted to developtechniques for isolation, purification, and culture of riceprotoplasts of cultivar IR64 and a wild rice species (Oryzaofficinalis). Optimization of protoplast isolation and purificationmethods from both rice genotypes were successfullydone. The highest protoplast density was obtained bydigesting embryonic callus or stems of young seedling in anenzyme solution containing of 2% cellulose, 0.1% pectolyase,0.5% macerozyme, 0.5% driselase, 5 mM ES, and 13% mannitolin CPW solution. The protoplast digestion was done forthree hours by soaking in the enzyme solution followed byshaking at 50 rpm under a room temperature. Purification ofthe protoplasts were done by separating them from plantdebris using a 25% sucrose solution. Protoplast regenerationwas not successful using although different media compositionsand conditions. Growth process from cell division tocell aggregate was only successful on IR64 protoplast cultureon a medium that contained AgNO3.
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机译：原生质体融合或体细胞杂交技术是用于生产植物杂交体的替代技术，所述植物由于其有性不相容性而难以通过常规方法生产。进行了一项实验，以开发用于分离，纯化和培养栽培品种IR64和野生稻种（Oryzaofficinalis）的水稻原生质体的技术。成功完成了两种水稻基因型原生质体分离和纯化方法的优化。通过在含2％纤维素，0.1％果胶酶，0.5％铜绿酶，0.5％果胶酶，5 mM ES和13％曼尼替林CPW溶液的酶溶液中消化幼苗的胚性愈伤组织或幼芽茎可获得最高的原生质体密度。原生质体消化通过浸泡在酶溶液中三个小时来完成，然后在室温下以50 rpm摇动。原生质体的纯化是通过使用25％蔗糖溶液将其与植物残渣分离来完成的。尽管培养基组成和条件不同，但原生质体再生并不成功。从细胞分裂到细胞聚集体的生长过程仅在包含AgNO3的培养基上进行IR64原生质体培养时才成功。
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