Development of PCR-ELISA for Detection and Differentiation of<i>Didymella bryoniae</i>from Related<i>Phoma</i>species

Somai, Benesh Munilal; Keinath, Anthony P.; Dean, Ralph A.

doi:10.1094/pdis.2002.86.7.710

Cited by 37 publications

(23 citation statements)

References 23 publications

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“…The 37 Brazilian isolates and the three US reference isolates of D. bryoniae were first tested with RG‐specific primers designed by Somai et al. (2002b).…”

Section: Methodsmentioning

confidence: 99%

Host Specificity and Genetic Diversity of Didymella bryoniae from Cucurbitaceae in Brazil

Santos¹,

Ferreira²,

Pessoa-Filho³

et al. 2009

Journal of Phytopathology

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Thirty-seven isolates of Didymella bryoniae from three Cucurbitaceae species were collected in Brazil and tested for pathogenicity to watermelon. All isolates were pathogenic but differed in aggressiveness levels. Seven representative isolates were used in cross-pathogenicity tests against 10 cucurbitaceous hosts. Most isolates were pathogenic to most host species tested, except to Sechium edule. Among the susceptible species, Citrullus and Cucumis species were the most susceptible hosts, while pumpkin and Luffa purgans were the most resistant. Host of origin affected the pattern of aggressiveness on each host. Isolates from watermelon were very aggressive to their original host, but much less aggressive or not pathogenic at all to some Cucurbita. Two previously described random-amplified polymorphic DNA (RAPD)-specific primers indicated that 81% of the isolates could be classified into the so-called RG I group, while the remaining isolates could not be classified into any of the described RG groups. All 37 isolates were further characterized by RAPD fingerprinting and compared with three US isolates representative of RG I and RG II groups. The Brazilian D. bryoniae isolates could be separated into genetically similar clusters. The majority of the isolates were grouped in cluster DB Ia, which contained only isolates of Citrullus lanatus and Cucumis melo. Two of the American isolates used as controls clustered with this group at 68% similarity level. The DB Ib cluster included three Brazilian isolates obtained from melon and watermelon and the American representative for RG II, at a lower similarity level (43%). Two isolates from watermelon clustered with one isolate from melon in a separate group (DB II), while one single isolate from pumpkin (DB III) showed the lowest genetic similarity to all other isolates. Didymella bryoniae isolates from Brazil showed, therefore, a level of genetic diversity higher than previously reported for the species. RAPD fingerprinting allowed for geographical distinction of D. bryoniae isolates but no correlation between genetic distance, aggressiveness or origin of the isolate was found.

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“…The 37 Brazilian isolates and the three US reference isolates of D. bryoniae were first tested with RG‐specific primers designed by Somai et al. (2002b).…”

Section: Methodsmentioning

confidence: 99%

Host Specificity and Genetic Diversity of Didymella bryoniae from Cucurbitaceae in Brazil

Santos¹,

Ferreira²,

Pessoa-Filho³

et al. 2009

Journal of Phytopathology

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“…Esca is induced by a complex of xylem-inhabiting fungi (Crous and Gams 2000), and there is evidence for early causal agents, which allow access to saprophytic opportunists. D. bryoniae often occurs with different Phoma species on cucurbit seedlings (Somai and Keinath 2002) or with powdery mildew (Zitter and Kyle 1992). Here we show that …”

Section: Discussionmentioning

confidence: 99%

Emerging multi-pathogen disease caused by Didymella bryoniae and pathogenic bacteria on Styrian oil pumpkin

et al. 2011

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The Styrian oil pumpkin, Cucurbita pepo L. subsp. pepo var. styriaca Greb. is a crop of cultural, commercial, and medical importance. In the last decade, yield losses of pumpkins increased dramatically. The ascomycetous fungus Didymella bryoniae (Fuckel) Rehm was identified as main causal agent provoking gummy stem blight as well as black rot of pumpkins. We observed a remarkable phenotypic diversity of the fungal pathogen, which contrasted with a high genotypic similarity. Evidence of pathogenictiy of D. bryoniae on Styrian oil pumpkin was demonstrated in a newly developed greenhouse assay. Isolates representing the five observed phenotypic groups fulfilled the Koch's postulates. In the field, the fungal disease was often associated with bacterial colonization by Pectobacterium carotovorum, Pseudomonas viridiflava, Pseudomonas syringae and Xanthomonas cucurbitae. The pathogenic behaviour of bacterial isolates on pumpkin was confirmed in the greenhouse assay. The high co-incidence of fungal and bacterial disease suggests mutualistic effects in pathogenesis. With a new assay, we found that bacteria can use the mycelium of D. bryoniae for translocation. We argue that the rapid rise of the multi-pathogen disease of pumpkins results from combined action of versatile pathogenic bacteria and the rapid translocation on a structurally versatile mycelium of the fungal pathogen.

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“…Detection of the fungus in seed lots may help farmers to manage the disease through rational sanitation efforts, such as seed dressing, and also discourage the commercial sale of contaminated seed. PCR-based assays to identify other species of Phoma have been developed (Somai et al 2002): P. macrostoma (Zhou et al 2005), P. sclerotioides (Larsen et al 2002), P. tracheiphila (Balmas et al 2005;Ezra et al 2007;Gentile et al 2000;Rollo et al 1990), P. ligulicola (Pethybridge et al 2004), P. foveata and P. exigua Lanes: M: PCR Marker; 1-2: P. betae control (−); 3-4: P. exigua control (−); 5-6: P. heteromorphospora control (−); 7-12: P. valerianellae; 13-15: CBS P. valerianellae control (+). Marker: GelPilot 1 Kb Plus Ladder (100) (Macdonald et al 2000).…”

Section: Discussionmentioning

confidence: 99%

Detection of Phoma valerianellae in lamb’s lettuce seeds

et al. 2010

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After the reappearance in Italy of a foliar disease of lamb's lettuce (Valerianella olitoria L. Poll. syn. V. locusta L. Betcke) incited by Phoma valerianellae, we set out to measure the level of seed infection by this fungus, using a plating test, and to develop a molecular method for quick and reliable detection of the pathogen in seeds. All six samples of lamb's lettuce seed tested were contaminated by P. valerianellae at levels of 0.6% to 15%. Surface disinfection of seeds did not eliminate the contamination and only reduced it to between 0.1% and 10%. The need for a sensitive, reliable and rapid diagnostic method for early identification of the fungus exists. We have developed a PCR-based method to identify the fungus in seeds. Variation within the internal transcribed spacer (ITS1, 5.8S sequences and ITS2) region of the rDNA (ITS) was used to characterize the P. valerianellae strains and to design specific primers within the ITS region.

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Development of PCR-ELISA for Detection and Differentiation ofDidymella bryoniaefrom RelatedPhomaspecies

Cited by 37 publications

References 23 publications

Host Specificity and Genetic Diversity of Didymella bryoniae from Cucurbitaceae in Brazil

Host Specificity and Genetic Diversity of Didymella bryoniae from Cucurbitaceae in Brazil

Emerging multi-pathogen disease caused by Didymella bryoniae and pathogenic bacteria on Styrian oil pumpkin

Detection of Phoma valerianellae in lamb’s lettuce seeds

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