Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol 8, Issue 4, 2006; Page No.(765-770)

PCR DETECTION OF HUANGLONGBING (GREENING DISEASE) IN SWEET ORANGE (CITRUS SINENSIS OSBECK)

K. GOPAL, V. GOPI, S. KHAYUM AHAMMED AND Y. SRINIVASULU

Abstract

Citrus greening disease (CGD) is caused by fastidious prokaryotic bacterium. The diseased trees show variety of symptoms. The diagnosis based on symptoms and biological indexing is often confused with symptoms of nutritional and physiological disorder. Survey data revealed that the mean CGD incidence was high in Prakasam district (26.91%) followed by Ananthapur (25.23%). Least mean incidence of 23.35% was recorded in Nalgonda district of Andhra Pradesh state of India. The CGB can be easily detected by a rapid and reliable method, PCR. Quality of DNA and its amplification was not satisfactory in PCR due to the presence of phenols and tannins in citrus tissue, which is responsible for degradation of DNA. Hence, two methods of DNA extraction were compared with CTAB method with respect to the quality of CGB DNA and its amplification in PCR. Addition of sodium sulphite to Tris-EDTA and CTAB method yielded good quality of DNA compared to Tris-EDTA method, which inturn resulted good amplification in PCR. Further, 6 ill CGB DNA was found good for better amplification in PCR.

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