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Disease Detection and Losses

Sensitive Broad-Spectrum Detection of Indian Peanut Clump Virus by Nonradioactive Nucleic Acid Probes. S. V. Wesley, Crop Protection Division, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, Andhra Pradesh 502 324, India, Current address Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, OK; J. S. Miller(2), P. S. Devi(3), P. Delfosse(4), R. A. Naidu(5), M. A. Mayo(6), D. V. R. Reddy(7), and M. K. Jana(8). (3)(4)(5)(7)Crop Protection Division, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, Andhra Pradesh 502 324, India; (2)(6)Virology Department, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, United Kingdom; Current address of J. S. Miller Agriculture Canada Research Station, Vancouver, BC V6T 1X2, Canada, and (8)Department of Agricultural Engineering, Indian Institute of Technology, Kharagpur, West Bengal 721 302, India. Phytopathology 86:1234-1237. Accepted for publication 15 July 1996. Copyright 1996 The American Phytopathological Society. DOI: 10.1094/Phyto-86-1234.

Cloned cDNA fragments representing different regions of the genome of the H serotype of Indian peanut clump furovirus (IPCV) were used as hybridization probes. Dot-blot assays using digoxigenin-labeled probes readily detected IPCV in extracts from infected plants with a sensitivity approaching that of using ³²P-labeled probes. The specificity of the probes depended on the part of the IPCV genome represented in the cDNA. Two probes representing the coat protein gene and adjacent gene in RNA-2 readily detected infection by H serotype IPCV, but reacted poorly with samples of plants infected with other serotypes. A probe corresponding to sequence near the 5’-end of RNA-1 reacted with samples of all IPCV serotypes, but not with those of peanut clump virus (PCV) from West Africa. A probe corresponding to the 3’-terminal 742 nucleotides of RNA-1 readily detected RNA of any IPCV serotype and of PCV. Assays using this probe, labeled with a nonradioactive reporter-digoxigenin, could detect IPCV in samples of seed tissue, as well as in tissue of several weed species and crop species such as wheat. The probe did not react with samples from plants infected by other furoviruses.