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Citrus gummosis, caused by Phytophthora is one of the important disease of citrus, causing huge loss of yield every year in India. Diagnosis based on visible symptoms is generally unreliable and, thus advanced diagnostic tools, molecular characterization has revolutionized the detection test, making it feasible to test large number of samples. Soil samples from the rhizosphere of heavily infected, symptomatic citrus plants, showing damping off, foot rot, root rot, gummosis, etc., were collected from four different locations of Vidarbha region, Maharashtra during JulyDecember in 2016-17 and Phytophthora was isolated from four different samples using leaf bait and soil suspension methods on selective PARPH-CMA medium. Four different morphological features were observed in the colonies when cultured on different media. The patterns of growth were stellate stiated, light rosette, cottony with slightly rossette and dense cottony structures on PARPH-CMA, CMA, V8 juice agar and PDA media, respectively. Molecular characterization was done with ITS primers. The PCR amplicons obtained from all the isolates were of expected size in all four isolates, because the specificity of primer combinations towards Phytophthora. No unspecific amplification was observed in any of the isolates, confirmed that above primer combinations, amplifies ITS region of Phytophthora only. The study conclusively revealed that Phytophthora nicotianae is the most prevalent species in citrus orchards of Vidarbha region, causing the widespread problem of citrus decline. To control the infection, proper hygiene and use of healthy planting material is necessary. To accomplish this, present investigation focus on the production of Phytophthora infection free, quality planting material, by screening both, scion source and rootstock through molecular methods before grafting.

Original article Morphological and molecular characterization of Phytophthora isolated from citrus orchards in Maharashtra