Isolation of Plant Pathogen - Xanthomonas

By Neil Singh and Osmond Rebello. Practical Supervisors- Dr. Bhole, Dr. N. Deshpande,.

Introduction: Organisms which cause disturbance in the rhythmical equilibrium of a plant with respect to structure or physiology or both and may lead to death or may reduce the economic value of the plant and its products are called plant pathogens. Our aim was to isolate the bacterial pathogen causing citrus canker.
Materials and Mathod: 1. The source of isolation was infected lemon fruit. 2. Alcohol, 0.1% sterile Hgcl solution, sterile water, glass rods sterile petri plates for washing and surface sterilization. 3. For isolation of plant pathogen the medium used is nutrient agar. The composition of the medium is as follows: Yeast extract 0.5g Peptone 1.0g NaCl 0.5g Distilled water 100ml Agar 2.0g pH of medium 7.0-7.2 The medium is autoclaved at 15 Ibs\sq.inch for 20 mins. A saline suspension is prepared by crushing the cankers. This suspension is used for streaking of NA plates which are then incubated at room temperature for 2-4 days
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Xanthomonas colonies produce copious amounts of extracellular slime - a complex polysaccharide composed of more than one type of sugar (a heteropolymer). It is termed xanthan and has important commercial uses as a gel that is stable at relatively high temperatures. About 20,000 tonnes of xanthan are produced industrially from X. campestris each year. It is used as a gelling and stabilising agent in salad dressings, ice creams, toothpastes, cosmetics, waterbased paints, etc., and also as a drilling lubricant in oil wells.

Results and Discussion: After 4 days sufficient growth was obtained on NA plates. When isolation of desired bacterium was carried out, yellow coloured, smooth and butyrous, mucoid colonies were obtained. Upon gram staining it was confirmed that these colonies consisted of rod shaped, gram negative bacteria. Since colonies of this bacterium are rather slow growing(3-5 days for ideal growth) and knowing the host of this bacterium(lemon tree- Citrus species), it can be inferred that the isolated bacterium is Xanthomonas axonopodis pv.citri . References: th edition, Stanier ,Ingraham, Wheelis, Painter. 1. General Microbiology, 5 2. Bergays Manual of Systematic Bacteriology, Second edition, Volume 2, The Proteobacteria.