Electron microscopy and sequencing of reverse transcriptase and ribonuclease H (RT/RNase H) region of Badnavirus genome from two banana cultivars: Poovan (triploid: AAB) and Safed velchi (diploid: AB), exhibiting leaf streak symptoms, confirmed the association of Banana streak OL virus (BSOLV). As per ICTV species demarcation threshold of 80 % identity in RT/RNase H region, both the isolates were identified as BSOLV. Rolling circle and end-to-end amplification showed the association of two short episomal BSOLV variants: BSOLV-IN1 and BSOLV-IN2 from Poovan and Safed velchi banana, respectively. The genome sizes of both isolates were 6,950 nucleotides long, but shorter than the typical BSOLV genome of 7,389 bp. Open reading frames (ORFs) 1 and 2 of shorter BSOLV isolates shared almost complete nucleotide identity (>99 %) to that of BSOLV. However, the ORF 3 (5,130 bp) and intergenic region (IGR), 886 bp, showed deletions compared with ORF 3 (5,499 bp) and IGR (956 bp) of BSOLV. In phylogenetic analysis for ORF 3 polyprotein, both the isolates clustered with BSOLV, Banana streak CA virus (BSCAV), and Sugarcane bacilliform GA virus (SCBGAV). Identical ORF 1, ORF 2, and the presence of all the conserved domains in short ORF 3 and promoter elements in IGR indicated that these isolates represent replicationally competent shorter variants of BSOLV. These two shorter-than-BSOLV genome sequences and two other identical banana streak virus sequences in GenBank (BSV-TRY; DQ859899 and BSV-GD; DQ451009) might have evolved due to error-prone reverse transcription and splicing or excision from the integrated sequences by homologous recombination in natural banana hybrids under field conditions.
In December 2002, bottlegourd (Lagenaria siceraria L.) plants grown as a commercial crop in Pune, India (western Maharashtra) showed severe mosaic, interveinal chlorosis, and leaf deformation that resulted in fern-leaf appearance and severe fruit distortion in approximately 70% of the plants. Crude sap of collected samples was used to mechanically inoculate uninfected glasshouse-grown bottlegourd plants that reproduced symptoms observed in the field. Sap extracts from these glasshouse infected bottlegourd plants were used to mechanically inoculate selected indicator hosts. Chlorotic local lesions were produced on Chenopodium amaranticolor, and systemic symptoms were produced on Benincasa hispida, Citrullus lanatus, Cucumis sativus, Cucurbita moschata, C. pepo, Luffa cylindrical, and Trichosanthes anguina. The virus was specifically identified with serological testing using direct antigen coating enzyme-linked immunosorbent assay. The virus reacted strongly to Zucchini yellow mosaic virus (ZYMV) antiserum and did not react to Papaya ring spot virus-P (PRSV-P), Cucumber mosaic virus (CMV), and Watermelon mosaic virus (WMV) antisera. Electron microscopic examination of leaf-dip preparation from infected plants showed flexuous filamentous particles (720 to 760 nm long) that are typical of potyviruses. Natural infection of bottlegourd by ZYMV has been reported in the Hawaiian Islands (1). To our knowledge, this is the first report of this potentially destructive virus in bottlegourd in India. Reference: (1) D. E. Ullman et al. Plant Dis. 75:367, 1991.
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