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ELECTRON MICROSCOPIC EVIDENCE FOR PURIFICATION OF COCONUT ROOT (WILT) DISEASE PHYTOPLASMA M. Mayilvaganan; J. J. Solomon
International Coconut Community Journal Vol 18 No 01 (2002): CORD
Publisher : International Coconut Community

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37833/cord.v18i01.353

Root (wilt) disease phytoplasma was purified from diseased coconut tissues using discontinuous Percoll gradient centrifugation method. Crude sap prepared from coconut was layered on a discontinuous Percoll gradient of 15, 30, 50 and 60%(v/v). After centrifugation at 20,000 g for 30 min, the turbid fraction formed on the top of 30% gradient in the diseased plant material was recovered, processed and fixed for electron microscopy. Electron microscopic examination of sections prepared from purified preparation of diseased plant material showed typical cells of root (wilt) phytoplasma with heterogeneous sizes and more or less spherical shape that are similar to those found in sieve elements of diseased tissues and salivary glands of infective (viruliforms) insect vectors. These purified phytoplasma bodies showed trilaminar membrane with internal materials of ribosome granules and DNA fibrils. However, the yield in terms of number of cells was fewer and in addition to intact bodies, free membranes and empty bodies lacking internal contents also were observed.

Phytoplasma diseases of coconut in India - root (wilt) and tatipaka diseases J. J. Solomon; L. Geetha
International Coconut Community Journal Vol 20 No 1 (2004): CORD
Publisher : International Coconut Community

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37833/cord.v20i01.384

Root (wilt) and Tatipaka diseases in India are caused by phytoplasma. Lethal yellowing disease in Caribbean region, Lethal yellowing like disease in west and east Africa and Coconut stem necrosis.in Sumatra and Peninsular Malaysia are phytoplasma induced diseases of coconut in other continents. Root wilt disease in India was first reported from Kerala in 1874.Tatipaka disease was first noticed in Andhra Pradesh after the 1949 cyclone. The progress made on various aspects including history, nature of spread, description of external & internal symptoms, etiology abiotic factors, biotic agents implicated in transmission of disease, culturing, chemotherapy, screening of coconut germplasm, breeding for disease resistance/tolerance and disease management is detailed.

Development of DAC Indirect ELISA for the Rapid Detection of Coconut Root (Wilt) Disease M. Sasikala; K. R. Chithra; J. J. Solomon; G. Rajeev
International Coconut Community Journal Vol 17 No 02 (2001): CORD
Publisher : International Coconut Community

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37833/cord.v17i02.350

Identification of root(wilt) disease-free palm is a basic requirement for evolving disease resistant/tolerant planting materials for the management of this phytoplasma induced malady. For this purpose, various types of ELISA were performed with different enzymes and their respective substrates to standardize the most suitable and sensitive one. Indirect ELISA, Protein A indirect ELISA and F(ab')2 indirect ELISA using Alkaline Phosphatase conjugate and DAC indirect ELISA using Horse raddish peroxidase conjugate were carried out. Of the various types of ELISA tried, DAC indirect ELISA has been found to be the best for the rapid detection of coconut root(wilt) disease. In this assay, crude leaf extracts and unfractionated polyclonal antiserum were employed as test antigen and specific antibody respectively. Higher specificity was observed with the addition of gelatin and ovalbumin in the extraction medium and overnight incubation of ELISA plate at 5°C after substrate addition. Antigen titre was found to be very high in spear leaves followed by the next outer leaf. Similarly, maximum antigen titre was observed during the early stage of the diseased palm. The test could be completed within 44h and in a single ELISA plate, 20 samples with three replications could be screened using microlitre quantities of the specific antibody.

Electron Microscopic Evidence for Purification of Coconut Root (Wilt) Disease Phytoplasma M. Mayilvaganan; J. J. Solomon
International Coconut Community Journal Vol 18 No 01 (2002): CORD
Publisher : International Coconut Community

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37833/cord.v18i01.353

Root (wilt) disease phytoplasma was purified from diseased coconut tissues using discontinuous Percoll gradient centrifugation method. Crude sap prepared from coconut was layered on a discontinuous Percoll gradient of 15, 30, 50 and 60%(v/v). After centrifugation at 20,000 g for 30 min, the turbid fraction formed on the top of 30% gradient in the diseased plant material was recovered, processed and fixed for electron microscopy. Electron microscopic examination of sections prepared from purified preparation of diseased plant material showed typical cells of root (wilt) phytoplasma with heterogeneous sizes and more or less spherical shape that are similar to those found in sieve elements of diseased tissues and salivary glands of infective (viruliforms) insect vectors. These purified phytoplasma bodies showed trilaminar membrane with internal materials of ribosome granules and DNA fibrils. However, the yield in terms of number of cells was fewer and in addition to intact bodies, free membranes and empty bodies lacking internal contents also were observed.

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