Abstract:

6 isolates of Xanthomonas campestris pv. Campestris collected by University of Warwick was used as the materials. The affect factors of AFLP were optimized, which were DNA extraction method, digestion time of Mse I/EcoR I, compositions of PCR reaction system and staining method, and an efficient system for X. campestris pv. Campestris AFLP analysis was established. The optimized factors were: the digestion system was 50 μL, the DNA template was 600 ng, the Mse I and EcoR I were both 5 U, the temperature was 37℃, reaction time was 4-6 h, pre-amplified products were diluted 10-fold for next selective amplification, the detection method was silver staining (AgNO3 content was 8 g/L and staining time was 15 min or so). The system was a powerful support for molecular level of genetic diversity in X. campestris pv. Campestris.