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      Use of RAPD, enzyme activity staining, and colony size to differentiate phytopathogenic Fuzarium Oxysporum isolates from Iran Translated title: Uso de RAPD, coloração de atividade enzimática e tamanho de colônia para diferenciação de isolados fitopatogênicos de Fusarium oxysporum obtidos no Irã

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          Abstract

          Fusarium oxysporum is a common soilborn plant pathogen with a worldwide distribution. Fusarium yellows disease of chickpea (Cicer arientinum) caused by F.oxysporum is one of the most destructive soilborn disease which is a major production constraint in chickpea-growing regions of Iran. Three laboratory methods "amplification of genomic DNA using random primers, enzyme activity staining, and colony size determination" have been used to discriminate between highly virulent (HV) and weakly virulent (WV) isolates of F. oxysporum. On the basis of colony size (a traditional morphological method) and the ability of isolates to produce pectic enzymes, five HV isolates were differentiated from three WV isolates. The HV isolates formed large colony (ranging from 10.1 to 12.6 mm in diameter) and showed the same enzyme pattern,while the WV isolates produced small colony (ranging from 5.8 to 7.8 mm in diameter) and had not detectable enzyme activity in the stained overlaying gel. Twelve arbitrary 10-mer primers were tested on these 8 isolates of F. oxysporum by Polymerase Chain Reaction (PCR). Cluster analysis of the data from the DNA amplification by Random Amplified Polymorphic DNA (RAPD), differentiated HV from WV isolates. The results obtained from RAPD test confirmed the classification of these eight isolates based on pathogenicity test, colony size, and enzyme activity staining into two groups (HV and WV).

          Translated abstract

          Fusarium oxysporum é um fitopatógeno amplamente difundido nos solos. A doença amarela do grão-de-bico (Cicer arietinum), causada por ele é destrutiva e um grande problema nas regiões produtoras do Irã. Três métodos de laboratório: amplificação aleatória de DNA genômico (RAPD), coloração de atividade enzimática e determinação das dimensões de colônia, foram usados para diferenciar isolados de Fusarium oxysporum altamente virulentos (HV) e fracamente virulentos (WV). Baseado no tamanho da colônia (método morfológico tradicional) e na capacidade dos isolados produzirem enzimas pécticas, cinco isolados HV foram diferenciados de três WV. Enquanto os isolados HV formaram colônias grandes (entre 10,1 e 12,6 mm de diâmetro) e apresentaram o mesmo padrão enzimático, os isolados WV produziram colônias pequenas (entre 5,8 e 7,8 mm de diâmetro) e não tiveram atividade enzimática detectável no gel de cobertura corado. Doze primers arbitrários foram testados nos 8 isolados de Fusarium oxysporum por reação de polimerase em cadeia (PCR). As análises dos dados da amplificação de DNA pelo método de amplificação aleatória do DNA polimórfico (RAPD) diferenciaram os isolados HV dos WV. Os resultados obtidos pelo teste RAPD confirmaram a classificação dos 8 isolados pelos testes de patogenicidade, tamanho da colônia e atividade enzimática de coloração nos dois grupos, HV e WV.

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          Characterization of the formae speciales of Fusarium oxysporum causing wilts of cucurbits by DNA fingerprinting with nuclear repetitive DNA sequences.

          The genetic relatedness of five formae speciales of Fusarium oxysporum causing wilts of cucurbit plants was determined by DNA fingerprinting with the moderately repetitive DNA sequences FOLR1 to FOLR4. The four FOLR clones were chosen from a genomic library made from F. oxysporum f. sp. lagenariae 03-05118. Total DNAs from 50 strains representing five cucurbit-infecting formae speciales, cucumerinum, melonis, lagenariae, niveum, and momordicae, and 6 strains of formae speciales pathogenic to other plants were digested with EcoRV and hybridized with 32P-labeled FOLR probes. The strains were clearly distinguishable at the formae specialis level on the basis of FOLR DNA fingerprints. Fifty-two fingerprint types were detected among the 56 strains by using all FOLR probes. These probes were used to infer phylogenetic relationships among the DNA fingerprint types by the unweighted pair group method using averages and parsimony analysis. The fingerprint types detected in each of the formae speciales cucumerinum, lagenariae, niveum, and momordicae were grouped into a single cluster. However, two different genetic groups occurred in the formae specialis melonis. The two groups also differed in pathogenicity: one group caused wilts of muskmelon and oriental melon, while the second was pathogenic only to muskmelon. The fingerprint types of different formae speciales pathogenic to plants other than cucurbits were distinguishable from one another and from the fingerprints of the cucurbit-infecting strains. These results suggest that the cucurbit-infecting formae speciales are intraspecific variants distinguishable at the DNA level and in their host range.
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            Mitochondrial DNA (mtDNA)-Relatedness among Formae Speciales ofFusarium oxysporumin the Cucurbitaceae

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              Pectic zymograms and taxonomy and pathogenicity of the Ceratobasidiaceae

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                bjm
                Brazilian Journal of Microbiology
                Braz. J. Microbiol.
                Sociedade Brasileira de Microbiologia (São Paulo )
                1678-4405
                December 2002
                : 33
                : 4
                : 299-303
                Affiliations
                [1 ] Razi University Iran
                Article
                S1517-83822002000400004
                10.1590/S1517-83822002000400004
                c89190c1-dc3b-4e30-9966-6fca9c57e47f

                http://creativecommons.org/licenses/by/4.0/

                History
                Product

                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1517-8382&lng=en
                Categories
                MICROBIOLOGY

                Microbiology & Virology
                Fusarium oxysporum,RAPD,colony size,enzyme activity staining,tamanho de colônia,coloração de atividade enzimática

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