Serological relationships and purification of bud necrosis virus, a tospovirus occurring in peanut (Arachis hypogaea L.) in India*

Annals of Applied Biology (Impact Factor: 2.15). 02/2008; 120(2):279 - 286. DOI: 10.1111/j.1744-7348.1992.tb03425.x

ABSTRACT SummaryA procedure for the purification of a tospovirus which causes bud necrosis disease (BND) of peanut in India is described. The virus contained three polypeptides of 78 kDa, 54 kDa and 31 kDa. In two ELISA procedures the virus failed to react with antisera to tomato spotted wilt virus (TSWV) obtained from different sources and with an antiserum to impatiens necrotic spot virus (INSV). Additionally, in reciprocal tests TSWV and INSV antigens failed to react with antiserum to the virus infecting peanut in India.In electro-blot immunoassay 54 kDa and 31 kDa polypeptides of the virus reacted with the homologous antiserum. None of the heterologous antisera reacted with any of the three viral polypeptides. On the basis of serological differences the virus that causes BND in India is distinct and therefore has been named bud necrosis virus (BNV). This serotype appears to be restricted to Asia.

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    ABSTRACT: The complete nucleotide sequence of the S RNA of peanut bud necrosis virus (PBNV) has been determined. The RNA is 3 057 nucleotides in length, contains inverted repeats and two open reading frames (ORFs) with an ambisense coding strategy that are separated by an A+U-rich intergenic region. One ORF (1 320 nucleotides in the viral sense strand) encodes a Mr 49.5 kDa protein, identified as the nonstructural (NSs) protein based on similarity to published tospovirus sequences. The second ORF (831 nucleotides in virus complementary strand) encodes a Mr 30.6 kDa protein. This protein was identified as the nucleocapsid (N) protein based on sequence similarities. Amino acid sequence comparison of N and NSs proteins revealed identities of 22-34% with the reported tospovirus isolates of serogroups I, II, and III, whereas it had 82-86% identity with viruses in serogroup IV, watermelon silver mottle virus (WSMV) and tomato isolate of peanut bud necrosis (PBNV-To). Two subgenomic RNA species detected in PBNV infected tissue corresponded to the predicted sizes (1.65 and 1.4 kb) of the NSs and N mRNAs. The data presented show conclusively that PBNV should be included in serogroup IV, along with WSMV and PBNV-To.
    Archives of Virology 02/1996; 141(1):85-98. · 2.28 Impact Factor
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    ABSTRACT: Genetic engineering of peanut (Arachis hypogaea L.) using the gene encoding for the nucleocapsid protein (N gene) of peanut bud necrosis virus (PBNV; genus Tospovirus, family Bunyaviridae) was used to impart resistance to bud necrosis disease in peanut (PBND), a disease for which no durable resistance is available in the existing germplasm. Over 200 transgenic lines of peanut var. JL 24 were developed for which integration and expression of the transgenes was confirmed by PCR, Southern hybridization, RT-PCR and western blot analysis. The T(1) and T(2) generation transgenic plants were assayed through virus challenge in the greenhouse by using mechanical sap inoculation at 1:100 and 1:50 dilutions of PBNV, and they showed varying levels of disease incidence and intensity. Greenhouse and field evaluation with T(2) generation plants indicated somewhat superior performance of the three transgenic events that showed considerable reduction in disease incidence. However, only one of these events showed over 75 % reduction in disease incidence when compared to the untransformed control, indicating partial and non-durable resistance to PBND using the viral N-gene.
    Archives of Virology 09/2012; · 2.28 Impact Factor
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    ABSTRACT: Tospovirus isolates from tomato and watermelon in Taiwan and from peanut in India reacted neither with antibodies to tomato spotted wilt virus (TSWV), nor with antibodies to impatiens necrotic spot virus (INSV), in ELISA and in electroblot immunoassays. Additionally, all three tospovirus isolates had an N protein with a molecular weight of 32 kDa as compared to 29 kDa for the N protein to TSWV and INSV. The observed physicochemical and serological differences suggest the generation of a new, third species in the genus tospovirus for which we propose the name groundnut bud necrosis virus.
    Archives of Virology 02/1993; 130(3-4):237-50. · 2.28 Impact Factor