preliminary identification and typing of pathogenic and toxigenic fusarium species using restriction digestion of its1-5.8s rdna-its2 region

Authors

h mirhendi

a ghiasian

hf vismer

mr asgary

abstract

background: fusarium species are capable of causing a wide range of crop plants infections as well as uncommon hu­man infections. many species of the genus produce mycotoxins, which are responsible for acute or chronic diseases in ani­mals and humans. identification of fusaria to the species level is necessary for biological, epidemiological, pathologi­cal, and toxicological purposes. in this study, we undertook a computer-based analysis of its1-5.8srdna-its2 in 192 genbank sequences from 36 fusarium species to achieve data for establishing a molecular method for spe­cie-specific identification. methods: sequence data and 610 restriction enzymes were analyzed for choosing rflp profiles, and subsequently de­signed and validated a pcr-restriction enzyme system for identification and typing of species.  dna extracted from 32 refer­ence strains of 16 species were amplified using its1 and its4 universal primers followed by sequencing and restric­tion enzyme digestion of pcr products. results: the following 3 restriction enzymes tas i, ita i and cfo i provide the best discriminatory power. using its1 and its4 primers a product of approximately 550bp was observed for all fusarium strains, as expected regarding the se­quence analyses. after rflp of the pcr products, some species were definitely identified by the method and some strains had different patterns in same species. conclusion: our profile has potential not only for identification of species, but also for genotyping of strains. on the other hand, some fusarium species were 100% identical in their its-5.8srdna-its2 sequences, therefore differentia­tion of these species is impossible regarding this target alone. its-pcr-rflp method might be useful for preliminary differ­entiation and typing of most common fusarium species.

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Journal title:
iranian journal of public health

جلد ۳۹، شماره ۴، صفحات ۳۵-۴۴

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