Radchuk V V, Klocke E, Radchuk R I, Neumann M
Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kiev, Ukraine.
Genetika. 2000 Jul;36(7):932-41.
The procedure for genetic transformation of two spring and one winter rapeseed cultivars was developed. No-paline strains of Agrobacterium tumefaciens GV3101 and EHA105 were shown to be preferable for gene transfer, as compared to the octopine strain GV2260. With two types of plant explants, the segments of hypocotyls and cotyledons, transformation was successful; however, its efficiency was somewhat higher with the fragments of hypocotyls. Analysis of regenerated plants by PCR and Southern blotting confirmed the presence of the nptII and nisA genes in transformants. RNA analysis by Northern blotting showed expression of the nisA gene in transformed shoots. The transgenes were inherited in T2 as Mendelian traits. The effect of biotic and abiotic factors on the efficiency of genetic transformation in rapeseed is discussed.
开发了两种春油菜品种和一种冬油菜品种的遗传转化程序。与章鱼碱型菌株GV2260相比，根癌农杆菌GV3101和EHA105的无章鱼碱菌株被证明更适合基因转移。使用两种类型的植物外植体，即下胚轴和子叶片段，转化成功；然而，下胚轴片段的转化效率略高。通过PCR和Southern印迹对再生植株进行分析，证实转化体中存在nptII和nisA基因。通过Northern印迹进行的RNA分析表明，nisA基因在转化芽中表达。转基因在T2代中作为孟德尔性状遗传。讨论了生物和非生物因素对油菜遗传转化效率的影响。
