Identification of Zucchini yellow mosaic potyvirus by RT-PCR and analysis of sequence variability

• Published in: Journal of virological methods Vol. 55; no. 1; pp. 83 - 96
• Main Authors: Thomson, K.G., Dietzgen, R.G., Gibbs, A.J., Tang, Y.C., Liesack, W., Teakle, D.S., Stackebrandt, E.
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01-09-1995; Amsterdam Elsevier; New York, NY
• Subjects: Amino Acid Sequence; Base Sequence; Biological and medical sciences; Capsid - genetics; Capsid Proteins; Cucurbita; DNA, Viral - analysis; Fundamental and applied biological sciences. Psychology; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control; Genes, Viral; Genetic Variation; Molecular Sequence Data; Nucleotide sequence variability; Phylogeny; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; Polymerase Chain Reaction; Potyvirus; Potyvirus - classification; Potyvirus - genetics; Potyvirus - isolation & purification; Restriction Mapping; Reverse transcription-polymerase chain reaction (RT-PCR); Sensitivity and Specificity; Sequence Analysis; Transcription, Genetic; Zucchini yellow mosaic virus; Australia; Oceania; Nucleotide sequence variability; Zucchini yellow mosaic virus; Potyvirus; Reverse transcription-polymerase chain reaction (RT-PCR); RNA-directed DNA polymerase; Genetic variability; Plant pathogen; Nucleotide sequence; Enzyme; Transferases; Cucurbitaceae; Identification; Method; Phylogeny; Infection; Virus; Polymerase chain reaction; Nucleotidyltransferases; Dicotyledones; Viral disease; Angiospermae; Spermatophyta; Potyviridae; Aminoacid sequence
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/0166-0934(95)00047-X

A reverse transcription-polymerase chain reaction (RT-PCR) method was used to identify Zucchini yellow mosaic virus (ZYMV) in leaves of infected cucurbits. Oligonucleotide primers which annealed to regions in the nuclear inclusion body (NIb) and the coat protein (CP) genes, generated a 300-bp product from ZYMV and also from the closely related watermelon mosaic virus type 2 (WMV-2). However, no product was obtained from papaya ringspot potyvirus which also infects cucurbits. ZYMV and WMV-2 were differentiated using a third primer which was complementary to a sequence in the 3′-untranslated region; a 1186-bp amplified product was obtained for ZYMV only. Nucleotide sequence analysis of the 300-bp fragments of Australian ZYMV and WMV-2 strains revealed 93.7–100% sequence identity between ZYMV strains. Multiple sequence alignments indicated that the nucleotide sequence which codes for the N-terminus of the CP was 74–100% identical for different isolates of ZYMV. The Australian isolate of WMV-2 was 43–46% identical to all isolates of ZYMV and was 84.6% identical to a Florida isolate of WMV-2.