Cervera M, Pina J A, Juárez J, Navarro L, Peña L
Dpto. Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Apartado Oficial, E-46113 Moncada, Valencia, Spain e-mail:
Plant Cell Rep. 1998 Dec;18(3-4):271-278. doi: 10.1007/s002990050570.
The effects of cocultivation with Agrobacterium tumefaciens, regeneration and selection conditions on the transformation efficiency of citrange (Citrus sinensis L. Osbeck×Poncirus trifoliata L. Raf.) have been investigated. Factors such as cocultivation period, preculture of explants, use of acetosyringone or feeder plates during cocultivation, cocultivation on a medium rich in auxins, postcultivation in darkness, and different kanamycin concentrations for selection were assessed. A 3-day cocultivation on a medium rich in auxins improved transformation frequencies, since it increased the number of dividing cells competent for transformation, at the cut ends of the explants. Exposure of explants to darkness for 4 weeks on selection medium resulted in further callus development and increased the regeneration frequency of transgenic shoots. Furthermore, this treatment drastically reduced the number of regenerated escape shoots. A transformation efficiency of 41.3% was achieved using the optimized transformation procedure.
研究了与根癌农杆菌共培养、再生和选择条件对枳橙（Citrus sinensis L. Osbeck×Poncirus trifoliata L. Raf.）转化效率的影响。评估了共培养时间、外植体预培养、共培养期间使用乙酰丁香酮或饲养层平板、在富含生长素的培养基上共培养、黑暗中继代培养以及不同卡那霉素浓度用于选择等因素。在富含生长素的培养基上进行3天共培养提高了转化频率，因为它增加了外植体切口端可用于转化的分裂细胞数量。外植体在选择培养基上黑暗处理4周导致愈伤组织进一步发育，并提高了转基因芽的再生频率。此外，这种处理大大减少了再生逃逸芽的数量。使用优化的转化程序实现了41.3%的转化效率。
