Fig. 2
Validation of the transformant strains, floral virulence test and stress evaluations. A To select transformants where the cassette was correctly inserted into the TSI locus 1, we amplified four different PCR products. Primer combinations P5–P6 and P9–P10 were used to verify the insertion event. Primer combination P7–P8 evaluated whether the recombination event between the two PCR fragments had been successful. Primer combination P11–P12 tested whether each transformant was homokaryotic for the transgene. Red asterisks indicate the expected PCR size bands. The transformant in lane 5 displayed a slightly higher band size for primer combination P9–P10 when compared to the other transformants. The higher band in lane 5 could be the product of unequal crossover in the RB region of the cassette. B Wheat spikes inoculated with PH-1 or the transformant strain (PtrpC-mCherry-TtrpC). No differences were observed in the number of infected spikelets showing typical disease symptoms. Photographs were taken at 12 dpi. Bars graph shows the number of infected spikelets between PH-1 and the transformant strain. Error bars indicate SD. C All the transformed strains showed a similar morphology and growth rate as the wild type PH-1 for all the conditions tested. Photographs were taken after 3 dpi. Salt stress (NaCl), membrane stresses (Calcofluor, Congo Red, Tergitol, SDS). PDA potato dextrose agar only