Vegetative compatibility and molecular characterization of Fusarium graminearum isolates from the State of Paraná, Brazil

Fusarium graminearum isolates causing Fusarium head blight in wheat were collected in Brazil and analyzed by random amplified polymorphic DNA (RAPD) markers and vegetative compatibility grouping (VCG). Nitrate non-utilizing mutants (nit) from each isolate were paired to verify heterokaryon formation. Three VCGs were identified among F. graminearum isolates: VCG1 included F-2, F-3 and F-4 isolates; VCG2 included F-1, F-6 and F-9 isolates; VCG3 included F-5, F-7 and F-8 isolates. Based on PCR amplification with eight different primers, the isolates showed great genetic similarity among themselves. Dendrogram analysis demonstrated two RAPD groups: Group A, consisting of isolates F-2 and F-9, and Group B, composed of the remaining isolates. Results suggest the clonal origin of F. graminearum isolates.

Isolados de Fusarium graminearum, obtidos de espigas de trigo com sintomas de Giberela, foram analisados pela técnica do Polimorfismo de DNA Amplificado ao Acaso (RAPD) e pelos Grupos de Compatibilidade Vegetativa (GCV). Mutantes auxotróficos (nit) de cada isolado foram pareados em todas as combinações possíveis, para a formação de heterocários. Três GCVs foram identificados: GCV1, incluindo os isolados F-2, F-3 e F-4; GCV2, incluindo os isolados F-1, F-6 e F-9; e GCV3, formado pelos isolados F-5, F-7 e F-8. Dois grupos foram identificados com base nos marcadores de RAPD: o grupo A, formado pelos isolados F-2 e F-9, e o grupo B, composto pelos demais isolados, os quais apresentaram grande similaridade entre si. Os resultados sugerem a origem clonal dos isolados de F. graminearum analisados.

Inibição do desenvolvimento de fungos fitopatogênicos por detergente derivado de óleo da mamona (Ricinus communis) / The castor oil plant detergent (Ricinus communis) inhibits the asexual development of phytopathogenic fungi

No presente estudo avaliou-se o efeito fungitóxico do detergente derivado do óleo da mamona (Ricinus communis) sobre o desenvolvimento dos fitopatógenos: Pyricularia grisea, Fusarium graminearum e Colletotrichum lindemuthianum. Seis concentrações do detergente (12,5mL L-1 a 300mL L-1) foram, individualmente, incorporadas ao Meio Basal; a seguir, após inoculação fúngica, o crescimento radial dos micélios foi avaliado. A inibição total do desenvolvimento de C. lindemuthianum e P. grisea foi observada entre as concentrações de 50mL L-1 e 200mL L-1, respectivamente. Com base no crescimento miceliano das colônias de F. graminearum, a atividade antifúngica do detergente do óleo da mamona (DOM) determinou inibição variável entre 79,4 e 91 por cento para a raça F2 e entre 80,7 e 90,7 por cento para a raça F4. O detergente, nas concentrações de 100 a 300mL L-1, inibiu em 100 por cento a germinação de conídios de F. graminearum (raças F-4 e F-2). Os resultados demonstram nítida atividade antifúngica do detergente derivado do óleo da mamona sobre fitopatógenos.

In the present study the fungitoxic effect of the castor oil plant detergent (Ricinus communis) on the development of the phytopathogens Pyricularia grisea, Fusarium graminearum and Colletotrichum lindemuthianum was evaluated. Six concentrations of the detergent (12.5mL L-1 to 300mL L-1) had been, individually, incorporated to the Basal Medium. After fungi inoculations, the radial growth of mycelia were evaluated. Detergent at 50mL L-1 and 200mL L-1 inhibited completely the development of P. grisea and C. lindemuthianum, respectively. On the basis of the mycelial growth of F. graminearum, the fungitoxic activity of the castor oil plant detergent (DOM) determined inhibition in the range of 79.4 and 91 percent for the F2 race and 80.7 and 90.7 percent for the F4 race. Detergent at the concentrations of 100mL L-1 to 300mL L-1 inhibited in 100 percent the F. graminearum germination conidia (races F-4 and F-2). Results demonstrate the fungitoxic activity of the castor oil plant detergent on phytopathogenic fungi.

Genotoxicity of the cyclo-oxygenase-inhibitor sulindac sulfide in the filamentous fungus Aspergillus nidulans

Sulindac sulfide is a non-steroidal anti-inflammatory drug (NSAID) with chemopreventive effect on human cancer cells. Due to the involvement of the somatic recombination in the carcinogenic process, sulindac sulfide's recombinogenic potential was evaluated by the Homozygotization Index (HI) in the filamentous fungus Aspergillus nidulans. The drug's recombinogenic potential was evaluated by its capacity to induce homozygosis of recessive genes from heterozygous diploid cells. Sulindac sulfide at 175 and 350 æM concentrations induced mitotic recombination in A. nidulans diploid cells, with HI values for genetic markers higher than 2.0, and significantly different from control HI values. The recombinogenic effect of NSAID was related to the induction of DNA strand breaks and cell cycle alterations. Sulindac sulfide's carcinogenic potential was also discussed.

Sulfeto de sulindaco é um antiinflamatório não-esteroidal com efeitos quimiopreventivos em cânceres humanos. O presente estudo teve como objetivo avaliar o potencial recombinagênico do sulfeto de sulindaco em células diplóides de Aspergillus nidulans. O efeito recombinagênico da droga foi demonstrado através da homozigotização de genes recessivos, previamente presentes em heterozigose. Os valores de HI (índice de Homozigotização) para diferentes marcadores genéticos apresentaram-se maiores do que 2,0 e significativamente diferentes dos valores obtidos em sulfeto de sulindaco ausência da droga (controle). O potencial recombinagênico do sulfeto de sulindaco foi associado à indução de quebras na molécula do DNA e a alterações no ciclo celular. O potencial carcinogênico do sulfeto de sulindaco foi discutido no presente trabalho.

Genotoxic evaluation of sodium nitroprusside in Aspergillus nidulans

The exogenous nitric oxide donor, sodium nitroprusside, evaluated the recombinogenic potential of nitric oxide. Drug inhibited mycelial growth and conidiation in A757 Aspergillus nidulans master strain. Two heterozygous diploid strains, one wild (uvsH+//uvsH+) and the other defective to DNA repair (uvsH//uvsH) were used for recombinagenesis tests. Sodium nitroprusside recombinogenic effect was evaluated by the induction of homozygosis of recessive genes, originally present in heterozygous condition. Results show that sodium nitroprusside (40 uM, 80 uM and 160 uM) is effective in inducing mitotic crossing-over in diploid cells of A. nidulans

Doxorubicin and etoposide induce somatic recombination in diploid cells of Aspergillus nidulans

Doxorubicin and etoposide are intercalating agents that inhibit the action of the enzyme topoisomerase II. Both drugs present therapeutic activity in numerous human neoplasms In the present work the recombinagenic potential of these drugs was evaluated by ascomycete Aspergillus nidulans. Their effects on the asexual cycle of A. nidulans was also appraised. Two heterozygous diploid strains of A. nidulans, a wild (uvsH+//uvsH+) and a defective to the DNA repair (uvsH//uvsH) were used. The drugs' recombinagenic potential was evaluated by their capacity to induce homozygosis of recessive genes from heterozygous cells. Both drugs have a recombinagenic effect on diploid cells of A. nidulans. Doxorubicin and etoposide are potentially capable to induce secondary malignancies, mediated by the mitotic crossing-over in eukaryotic cells

Vincristine induces somatic segregation, via mitotic crossing-over, in diploid cells of Aspergillus nidulans

Vincristine is an alkaloid widely used as an antineoplastic agent. In eukaryotic cells the drug causes blockage in the G2 phase of the cell cycle and an increase in the frequency of sister chromatid exchanges. Due to the fact that germinating Aspergillus nidulans cells spend most of their cycle in G2 phase, they provide an excellent system for the study of mitotic crossing-over. Taking into account that mitotic crossing-over occurs during G2 period, the evaluation of recombinagenic and aneugenic potential of vincristine is provided with regard to two diploid strains of A. nidulans: a wild strain (uvsH+//uvsH+) and a defective one in DNA repair (uvsH//uvsH). Drug toxicity and its effect on the asexual cycle of A. nidulans has been evaluated as well. Treatment of both strains with vincristine did not change colony growth in the culture, however cytological analyses showed aberrant conidiophores. Recombinagenic potential of vincristine was evaluated by induction of gene homozygosis originally present in heterozygosity diploid strains (Homozygotization Index). Results show that vincristine induces mitotic crossing-over and higher frequency of aneuploid mitotic segregants. The results also show the recombinagenic and aneuploidogenic potential of vincristine and suggest its participation in the induction of secondary malignancies.

uvsZ1 mutation shows epistatic relations with uvsD153 and uvsJ1 mutations without any involvement with checkpoint control in Aspergillus nidulans

The participation of the recently described uvsZ1 mutation in checkpoint control and the identification of epistatic relations between uvsZ1 mutation and uvsD153 and uvsJ1 mutations are provided. The effect of mutation uvsZ1 in mitotic exchanges into paba-bi (chromosome I) and cho-nic (chromosome VII) genetic intervals has also been evaluated. The mutation uvsZ1 was epistatic with regard to uvsD153 and uvsJ1 mutations, with no involvement with checkpoint control. In contrast to mutations in UvsB and UvsF groups, the uvsZ1 mutation failed to cause any changes in the frequencies of mitotic crossing-over. The distinct phenotypic traits given by mutation uvsZ1 suggest the presence of complex interactions among the different DNA repair pathways. Interaction may be an additional cell strategy of DNA damage response

Characterization and mapping of an informational suppressor in Aspergillus nidulans

The present work was undertaken to characterize a suppressor gene present in a mutant strain of A. nidulans obtained with NTG (N-Methyl-N'-Nitro-N-Nitrosoguanidine). Analyses of this mutant have shown that this suppressor, designated suO1, induces phenotypic co-reversion of several auxotrophic mutations and makes the strain sensitive to aminoglycoside antibiotics and lower temperatures. suO1 has shown to be on linkage group VIII. The vegetative growth of the mutant strain is very unstable because the suppressor gene induces the production of prototrophic mitotic sectors. The strains bearing the suO1 gene produce cleistothecia containing a reduced number of viable ascospores during the sexual cycle. The segregation of the genetic markers has also been observed in the mutant strain self crossed. From the above results it may be concluded that suO1 is an informational suppressor.