CSIR-<strong>IHBT</strong> <strong>Annual</strong> <strong>Report</strong> 2010-11Identification of plant proteins interacting with viral proteinsCucumber mosaic virus movement protein was found to interact with ascorbate oxidase gene ofplant and methyltransferase gene of virus. Similarly, RNA dependent RNA polymerase geneshowed interaction with ascorbate oxidase. In BLAST analysis, sequence showed similarity to 16sribosomal gene of Cucumis sativus.Development of diagnostics and molecular characterization of viruses infectingplum and cherry (Funded by Department of Biotechnology, Govt. of India)A multiplex RT-PCR was developed for the simultaneous detection of four major cherry virusesviz., Cherry virus A (CVA), Cherry necrotic rusty mottle virus (CNRMV), Prunus necrotic ringspot virus(PNRSV) and Little cherry virus -1 (LChV-1) infecting sweet cherry. The genome of CNRMVwas approximately 8.4 kb with 7 ORFs excluding the poly (A) tail. Five of these ORFs wereconserved among all fovea-, allexi-, potex and carlaviruses and coded for a methyltransferase/helicase/polymerase polypeptide, the triple gene block movement proteins and the coat protein.Two further ORFs, 2a and 5a, were completely nested within ORFs 2 and 5, respectivelydifferentiating it from Foveavirus and Potexvirus. A region of 2203 nt was sequenced and submittedto the GenBank. The region covers ORF2, ORF2a, ORF3, ORF4, ORF5, ORF5a and 3’-UTRregions of the CNRMV genome (Table 22).Table 22 Products coded by the different ORFs in the genome of CNRMVGene Protein Size (kDa)ORF2 Triple gene block 1 25ORF2a Unknown protein 13ORF3 Triple gene block 2 12ORF4 Triple gene block 3 7ORF5 Coat Protein 30ORF5a Unknown protein 16Complete genome of Cherry virus A (CVA)Sweet cherry (Prunus avium L.) is an important deciduous temperate fruit crop in the WesternHimalayan region of India. In order to determine the health status of cherry plantations and theincidence of CVA, orchards in the states of J&K and H.P. were surveyed. The incidence was 28and 13% from J&K and H.P., respectively. Complete genome was amplified and amplicon of about7.4kb was cloned and sequenced (7379bp). The genome organization was similar to that of isolatescharacterized earlier, coding for two ORFs, in which ORF2 was nested in ORF1. The completesequence was 81% similar to that of the type isolate with 5' and 3' UTRs of 54 and 299 nucleotides,respectively.65
CSIR-<strong>IHBT</strong> <strong>Annual</strong> <strong>Report</strong> 2010-11Bacterial diversity and soil enzyme activityA culture independent survey of the bacterial diversity in rhizosphere soils of diseased (scab, S2)and disease free (S14) apple trees was conducted to assess their role in disease suppressiveness. Thephylotypes and their frequency distribution in cloned libraries indicated that the phylotype didnot represent a single group. Rarefaction curve (Fig. 59), Shannon and Simpson diversity indicesexhibited insignificant differences between the samples in bacterial community composition.Chitinase and β-1,3 glucanase activities (Fig. 60) were higher in samples from disease free trees ascompared to scab infected ones. In disease free and diseased rhizospheres, dominance of unculturedbacteria was 70 and 72.5%, respectively in partially sequenced representative 80 clones.Fig. 59 Rarefaction curve showing differnce in type richness ofthe librariesFig. 60 Antifungal enzyme activities in soil suspensiona) chitinase and b) β-1,3-glucanaseSelection and differentiation of Bacillus spp. antagonistic to Fusarium oxysporumf.sp. lycopersici and Alternaria solani infecting tomatoIn dual culture assays, antagonistic Bacillus spp. displaying in vitro production of siderophore,chitinase and β-1,3-glucanase were identified. In greenhouse studies, seed bacterization andsoil application of B. atrophaeus S2BC-2 challenge inoculated with Fusarium oxysporum f.sp.66
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