Агробактеріальна трансформація in planta пшениці озимого сорту Подолянка та ярого сорту Bobwhite

Abstract

Aims. Triticum aestivum L., being one of the world’s most important staple crops, remains a challenge for genetic transformation. This paper describes the application of in planta method to obtain transformants by treating cells of intact plants with argobacterium. Methods. Analysis of the plant material for the presence of nptII transgene was carried out using polymerase chain reaction (PCR). Results. Fresh agrobacterium containing vector p014 was transferred into spikes of wheat. There were 13 seeds (18.0%) collected of cv. Bobwhite out of 72 possible from 12 spikes and 293 seeds (46.4%) of cv. Podolyanka out of 620 possible from 24 spikes. Based on the results of PCR for the presence of nptII transgene there were no positive signals found for the cv. Bobwhite. On the other hand there were 4 clear positive samples detected for cv. Podolyanka indicating the presence of nptII sequence. Conclusions. Agrobacterium-mediated in planta transformation was conducted for winter cv. Podolyanka and spring cv. Bobwhite. The ambient temperature below 28°C during the next 18 hours after agrobacterium application was crucial. Tested probes of cv. Podolyanka (13.3%) were positive for the presence of transgene nptII.

Key words: wheat, transformation, nptII, agrobacterium, GFP.