Abstract
DNA sequences, specific to each of the two taxa of Phomopsis (taxon 1 and taxon 2) that commonly infect grapevines in Australia, have been identified and cloned. The clones recognise a repeated sequence and both identify genetic variation. The taxon 2-specific marker identified five distinct banding patterns amongst twelve Phomopsis taxon 2 isolates obtained from five different geographical regions. The Phomopsis taxon 1-specific clone was used to detect the fungus in canes and unburst buds of grapevines that displayed symptoms of infection, visible in late winter as pycnidia on bleached canes. However, the fungus was also detected in canes and unburst buds from canes that were not bleached. In addition, perithecia of the sexual phase were observed on other unburst buds following incubation of canes. The results suggest that bleaching should not be used as an indicator of infection by Phomopsis taxon 1. Microscopic examination of infected, micropropagated grapevines showed that, similar to other Phomopsis species, taxon 1 grows subcuticularly in the stems. The specificity and high copy number of the taxon 1 and taxon 2 probes, together with a rapid DNA extraction method, provide the means for a sensitive assay for these fungi and for fundamental studies on the biology of two Phomopsis taxa that infect grapevine in Australia.
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References
Daykin ME, Milholland RD (1990) Histopathology of blueberry twig blight caused by Phomopsis vaccinii. Phytopathology 80, 736–740.
Evans KJ, Whisson DL, Scott ES (1996) An experimental system for characterizing isolates of Uncinula necator. Mycological Research 100, 675–680.
Evans KJ, Whisson DL, Stummer BE, Scott ES (1997) DNA markers identify variation in Australian populations of Uncinula necator. Mycological Research 101, 923–932.
Hewitt WB, Pearson RC (1988) Phomopsis cane and leaf spot. In ‘Compendium of grape diseases’. (Eds RC Pearson, AC Goheen) pp. 17–18. (APS Press: Minnesota, USA)
Hood WE, Shew HD (1996) Applications of KOH-aniline blue fluorescence in the study of plant-fungal interactions. Phytopathology 86, 704–708.
Igoe MJ, Peterson NC, Roberts DL (1995) A Phomopsis canker associated with branch dieback in Colorado blue spruce in Michigan. Plant Disease 79, 202–205.
Kuo K, Leu L-S (1998) Phomopsis vitimegaspora: a new pathogenic Phomopsis from vines. Mycotaxon 66, 497–499.
Meijer G, Megnegneau B, Linders EGA (1994) Variability for isozyme, vegetative compatibility and RAPD markers in natural populations of Phomopsis subordinaria. Mycological Research 98, 267–276.
Merrin SJ, Nair NG, Tarran J (1995) Variation in Phomopsis recorded on grapevine in Australia and its taxonomic and biological implications. Australasian Plant Pathology 24, 44–56.
Mostert L, Crous PW, Kang J-C, Phillips AJL (2001) Species of Phomopsis and a Libertella sp. occurring on grapevines with specific reference to South Africa: morphological, cultural, molecular and pathological characterization. Mycologia 93, 146–167.
Phillips AJL (1999) The relationship between Diaporthe perjuncta and Phomopsis viticola on grapevines. Mycologia 91, 1001–1007.
Pine TS (1959) Development of the grape dead-arm disease. Phytopathology 49, 738–743.
Pseicht JW, Pearson RC (1989) Time of infection and control of Phomopsis fruit rot of grape. Plant Disease 73, 829–833.
Rehner SA, Uecker FA (1994) Nuclear ribosomal internal transcribed spacer phylogeny and host diversity in the coelomycete Phomopsis. Canadian Journal of Botany 72, 1666–1674.
Rezaian MA, Krake LR (1987) Nucleic acid extraction and virus detection in grapevine. Journal of Virological Methods 17, 277–285.
Sambrook J, Fritsch EF, Maniatis T (1989) ‘Molecular cloning: a laboratory manual (2nd edn).’ (Cold Spring Harbor, New York)
Scheper RWA, Crane DC, Whisson DL, Scott ES (2000) The Diaporthe teleomorph of Phomopsis taxon 1 on grapevine. Mycological Research 104, 226–231.
Scheper RWA, Whisson DL, Scott ES (1997) Revised disease cycles for the two types of Phomopsis on grapevine. The Australian Grapegrower and Winemaker 405, 41–44.
Taylor RH, Mabbitt JM (1961) Dead-arm disease of grapevines. The Journal of the Department of Agriculture, Victoria, Biology Branch Publication 901, 157–165.
Thomas MR, Matsumoto S, Cain P, Scott NS (1993) Repetitive DNA of grapevine: classes present and sequences suitable for cultivar identification. Theoretical and Applied Genetics 86, 173–180
Velicheti RK, Lamison C, Brill LM, Sinclair JB (1993) Immunodetection of Phomopsis species in asymptomatic soybean plants. Plant Disease 77, 70–73.
Williamson PM, Sivasithamparam K, Cowling WA (1991) Formation of subcuticular hyphae by Phomopsis leptostromiformis upon latent infection of narrow-leafed lupins. Plant Disease 75, 1023–1025.
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Melanson, D.L., Rawnsley, B. & Scheper, R.W.A. Molecular detection of Phomopsis taxa 1 and 2 in grapevine canes and buds. Australasian Plant Pathology 31, 67–73 (2002). https://doi.org/10.1071/AP01075
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DOI: https://doi.org/10.1071/AP01075