摘要
Protoplasts of Valencia sweet orange ( Citrus sinensis Osb.),irradiated by X_ray with a dose rate of 3.8 krad/min for 45 min, were electrically fused with protoplasts of Murcott tangor ( C. reticulata×C. sinensis ) that were treated with 0.25 mmol/L iodoacetic acid for 15 min. It took nearly 15 months for the fusion_derived calli to develop into embryoids that were only originated in the medium of MT supplemented with 2% glycerol. The shoots were recalcitrant to rooting in the root_induction medium. In vitro grafting was employed to produce whole plants though one self_rooting plant was obtained. Cytological determination of root and shoot tips showed mainly diploid and aneuploid cells, together with few tetraploid cells in some plants. RAPD (random amplified polymorphism DNA) analysis with 10_mer primers demonstrated that bands specific to the fusion parents were detected in the regenerated plants, indicating that interspecific somatic hybrids have been obtained via protoplast asymmetric fusion in Citrus .
用 3.8krad/min的X射线辐射伏令夏甜橙 (CitrussinensisOsb .)原生质体 45min ,0 .2 5mmol/L碘乙酸处理Murcott橘橙 (C .reticulata×C .sinensis)原生质体 15min ,电场诱导两种原生质体融合。融合再生的愈伤组织培养近15个月后再生出胚状体 ,而且只能在MT +2 %甘油的培养基中再生。根尖和茎尖染色体检查表明 ,在再生植株中主要是二倍体细胞和非整倍体细胞 ,只在部分植株中观察到少数四倍体细胞。采用 10_mer随机引物进行RAPD分析 ,在再生植株中都能检测到融合亲本的特征带 ,表明通过非对称融合获得了柑橘种间体细胞杂种植株。
基金
TheNational Natural Science Foundation of China(39830260
39625017)
the International Foundation for Science(D/3001_1)
