Citrus is an important fruit grown all over the world, but it is vulnerable to attack of Xanthomonas axonopodis pv. citri (Xac), causative agent of citrus canker. It is documented as most devastating disease leading to great economic losses by affecting overall fruit production in citrus. In contemporary study Xac was isolated from the infected plant samples which were collected from Bahawalpur by using nutrient agar medium. The bacterium was purified on nutrient agar medium by streaking method. Confirmation of Xac was done by its morphological, biochemical and cultural features. It showed positive result in KOH solubility test, starch hydrolysis test, casein hydrolysis, catalase test, and citrate utilization test while it expressed negative result in fluorescent pigmentation, Staining test, indole production and Kovac’s oxidase test. The bacterium colonies appeared as mucoid, convex, yellow to orange color on NA medium; flattened or slightly raised, bright to pale yellow color, mucoid colonies on YDCA medium. This study is helpful for researchers and scientist for accurate identification of Xac which is necessary for in-vitro evaluation of different chemicals to find out most affective chemical against canker disease in field.