Petri César, Wang Hong, Alburquerque Nuria, Faize Mohamed, Burgos Lorenzo
Mejora de Plantas, CEBAS-CSIC, Aptd. de Correos 164, 30100, Murcia, Spain.
Plant Cell Rep. 2008 Aug;27(8):1317-24. doi: 10.1007/s00299-008-0550-9. Epub 2008 May 1.
A protocol for Agrobacterium-mediated stable transformation for scored, whole leaf explants of the apricot (Prunus armeniaca) cultivar Helena was developed. Regenerated shoots were selected using a two-step increased concentrations of paromomycin sulphate. Different factors affecting survival of transformed buds, including possible toxicity of green fluorescent protein (GFP) and time of exposure to high cytokine concentration in the regeneration medium, were examined. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines was 5.6%, when optimal conditions for bud survival were provided. Southern blot analysis on four randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene. This is the first time that stable transformation of an apricot cultivar is reported and constitutes also one of the few reports on the transformation of Prunus cultivars.
开发了一种用于杏(Prunus armeniaca)品种海伦娜带评分的全叶外植体的农杆菌介导稳定转化方案。使用两步增加硫酸巴龙霉素浓度的方法筛选再生芽。研究了影响转化芽存活的不同因素,包括绿色荧光蛋白(GFP)的潜在毒性以及在再生培养基中暴露于高细胞分裂素浓度的时间。当提供芽存活的最佳条件时,基于对来自独立株系的单个推定转化芽进行PCR分析,转化效率为5.6%。对四个随机选择的PCR阳性芽进行的Southern印迹分析证实了nptII转基因的存在。这是首次报道杏品种的稳定转化,也是关于李属品种转化的少数报道之一。