籼稻YTB成熟胚愈伤组织继代培养条件试验

Study on subculture conditions of YTB mature embryos of indica rice callus

为确定籼稻YTB成熟胚愈伤组织继代培养的最佳条件,对籼稻愈伤组织进行不同继代培养条件优化组合试验,研究不同因素对籼稻愈伤组织继代培养的影响。结果表明:NB培养基是较适合籼稻愈伤组织继代生长的培养基;籼稻YTB成熟胚愈伤组织不适合二次继代;铁盐浓度对籼稻愈伤组织的继代培养有明显影响;水解酪蛋白、脯氨酸、谷氨酰胺及硫代硫酸钠等能改善籼稻愈伤组织的质量,降低褐化率;抗坏血酸(VC)对籼稻胚愈伤组织的继代生长及抗褐化无明显影响;形态学观察和细胞学观察的一致性证明了外源腐胺PUT(30 mg/L)在籼稻愈伤组织继代培养中可以使愈伤组织保持良好的胚性,从而提高后续遗传转化的转化率。

To establish the most optimal callus subculture system on YTB mature embryo, the effect of different callus subculture conditions on YTB mature embryo were studied. The resuhs showed that NB medium was more suitable to the growth of callus subculture; YTB callus from mature embryos wasn＇t suitable for the second subculture. Fe iron concentration had a significant impact on callus subculture; hydrolysis of casein protein, proline, glutamine and sodium thiosulfate improved the quality of callus and reduced the browning rate; ascorbic acid （VC） had no significant impact on the embryo callus subculture growth and anti-browning. The consistency of morphology and cytology observation proved that exogenous putrescine （30 mg/L） in the callus subculture maintained callus in a good embryogenic condition, thereby enhanced the subsequent genetic transformation rate.