Morpho-Molecular Characterization of Xanthomonas Axonopodis Pv. Citri Associated with Kinnow (Mandarin) and its Management
Muhammad Iqbal1, Muhammad Ehetisham ul Haq1*, Muhammad Kamran1, Muhammad Idrees1, Shahid Nazir2, Ihsan Ullah3, Sumera Naz1, Shaukat Ali1 and Muhammad Zafar Iqbal2
ABSTRACT
Millions of dollars have been spent annually on quarantines, prevention, eradication and management programs around the globe. The present study was coined to record the incidence of the disease in the Sargodha districts of the Punjab province. Furthermore, isolated bacterium from diseased samples was characterized morphologically and molecularly. The relative efficacy of commercially available antibiotics against the pathogen was evaluated in the lab and in field conditions. A systematic survey of randomly ten localities/sampling sites was conducted for a reliable estimation of citrus canker disease in District Sargodha of Punjab Province. Nine sampling sites showed 90 % the presence of Xanthomonas axonopodis pv. citri and only one site was free of citrus canker disease. Total cellular DNA was isolated from freshly growing aqueous bacterial culture using CTAB protocol with modifications. Amplification of 581bp fragment from isolated DNA from bacterial isolates confirmed the presence of pathogen i.e. X. axonopodis pv. citri. The amplified PCR product was purified and further confirmed by sequencing which showed 100% similarity with 40 nucleotide sequences of X. axonopodis pv. citri submitted in NCBI. Relative efficacy of six antibiotics (Streptomycin sulphate, Oxytetracycline, Cefalexin, Kasugamycin, Kanamycin and Levofloxacin USP) against colony growth of X. axonopodis pv. citri was evaluated at 50, 100 and 150 ppm using disc sensitivity technique. Streptomycin sulphate was found most effective to inhibit the colony growth of the bacterium. Streptomycin sulphate and Oxytetracycline were evaluated against the disease in field conditions. Streptomycin sulphate was found relatively most effective as compared to the Oxytetracycline.
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