ANTISERUM PRODUCTION AND IMMUNOELECTRON MICROSCOPY OF POTATO LEAFROLL VIRUS
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Authors
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M. N. SINGH, S. M. PAUL KHURANA AND I. D. GARG
Central Potato Research Institute, Shimla 171 001
Keywords:
Potato leafroll virus, Luteovirus, Purification, ISEM, AntiserumAbstract
Potato leafroll virus (PLRV), a phloem limited luteovirus, was successfully purified with and without enzyme 'Driselase' or 'Celluclast' for macerating plant tissue from fresh infected P. floridana leaves/stems. The virus was extracted in 0.5 M potassium phosphate buffer (pH 7.4), clarified with chloroform and n-butanol, precipitated with polyethylene-glycol, resuspended and concentrated by differential centrifugation and finally purified by density gradient sedimentation. Yield of purified PLRV was almost doubled with the enzyme. Titre of the antiserum produced using the best virus preparation was 1 : 512. The virus was detected by IEM from single aphids.
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