Development of species-specific primers for detection of Xanthomonas campestris pv. campestris causing black rot of crucifers

Aim: Black rot of crucifers caused by Xanthomonas campestris pv. campestris (Pammel) Dowson (Xcc) is a major seed-borne disease. The present study aimed to develop a rapid diagnostic protocol for the specific and sensitive detection of this pathogen. Methodology: A specific primer set was designed based on rpf gene and optimization of PCR condition was done for specific detection of Xcc. Sensitivity of PCR for primer set was then determined by diluting the Xcc DNA and cells. Results: Specific primer set was able to amplify a specific band of 304 bp in all 11 isolates of Xcc but failed to amplify other Xanthomonas species and one each of Ralstonia solanacearum, Erwinia caratovora subsp. caratovora, Bacillus subtilis, Pseudomonas fluorescens and P. aeruginosa. The primer set was highly sensitive as it was able to detect 10 pg μl-1 bacterial DNA and up to 3x103 CFU ml-1 corresponding to 12 viable cells of Xcc which were used as template for PCR reaction Interpretation: The results suggest that developed PCR primers are highly specific and sensitive and it can be used to detect the pathogen at an early stage of infection for disease management.

Effect of Shading on Incidence of Black Rot Disease and Yield Quality of Broccoli in Humid Tropical Regions of Thailand.

The study was carried out in the experimental location of Department of Plant Science, Faculty of Natural Resources, Prince of Songkla University, Thailand, from April to June, 2012. Split-plot in randomized complete block design was used in this experiment with four replications. The broccoli plants under the shading had the lowest incidence of black rot disease (14.48%) which was significantly lower than the full sunlight (76.91%). There was a positive impact of the shading on yield quality of the broccoli. The Yok Kheo had the highest head length and diameter (39.13 cm and 12.11 cm, respectively) when planting under the shading. These results consistent with broccoli total yield, the highest total yield was obtained from the Yok Kheo under the both shading and full sunlight (10.92 t.ha-1 and 8.29 t.ha-1, respectively), followed by the Green Queen under the shading (6.21 t.ha-1). Therefore, the appropriate variety for planting during dry season in humid tropical regions of Thailand was the Yok Kheo and Green Queen because its total yield per hectare was higher than of the Top Green and they also had higher quality of head.

Artificial Screening for Black Rot Resistance Based on Different Disease Parameter in Early Cauliflower

India has maximum genetic materials in early cauliflower, which grow in subtropical conditions. Different disease parameters like linear growth, maximum growth rate per day, AUDPC, apparent infection rate and percent diseased area were calculated in artificially inoculated plants. Apparent infection rate is not co-related with the black rot disease incidence and should never be considered during characterization of disease resistance and varietal screening. Based on the above disease parameters Kunwari-18, Phool Gobhi Kunwari, Kataki-7 and BT-10-2 were selected as moderately resistance to black rot in early cauliflower. These lines can be used for black rot prone area and also for black rot disease improvement programme. Considering the qualitative and quantitative parameters, slow rotting resistance cauliflower lines are selected as such for cultivation and would be best suited in integrated disease programme.