Isolation of Aflatoxin-producing Species Based on Detection of afl R and ver 1 Genes and Sspecies Identification by Determination of ITS Sequences in Edible Rice of Mashhad, Khorasan
Subject Areas :Hamed Faraji 1 , Farideh Tabatabaee Yazdi 2 , Nematollah Razmi 3
1 - دانشجوی دکتری، گروه میکروبیولوژی، دانشکده کشاورزی و فن آوری مدرن ، واحد شیراز، دانشگاه آزاد اسلامی، شیراز، ایران.
2 - استاد، گروه علوم وصنایع غذایی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران.
3 - استاد، گروه میکروبیولوژی، دانشکده کشاورزی و فن آوری مدرن ، واحد شیراز، دانشگاه آزاد اسلامی، شیراز، ایران.
Keywords: Aflatoxin, Aspergillus, PCR,
Abstract :
Rice is an important cereal and also one of the essential sources of energy in the world and especially in the eastern world. Because of nutritional value and relative humidity, rice is certainly exposed able to contamination with molds. Aflatoxin is a type of secondary metabolites and one of the most dangerous and important fungal toxins. Aflatoxigenic Aspergillus species can be very terrible in cereal and dried fruits. Aspergillus flavous and A.parasiticus are two famous and most known species responsible for the contamination of food and feed stuff. Culturing and morphological characteristics were used in identification and isolation of Aspergillus isolates. Then molecular evaluation of the gene involves in the aflatoxin biosynthesis process was done by PCR. ver 1, aflR are structural and regulatory gene that has a critical role in aflatoxin producing system. Toxin production of isolated species gets confirmed by HPLC and Immunoaffinity column method. Results showed 20 samples of 60 have aflatoxigenic fungi. 2 types of fungi with similar morphological characteristics of Aspergillus flavous and A. parasiticus were identified and cultured on AFPA (specific culture media) for verification. PCR molecular test and cultural morphology were accepted by together in 90 percent of samples. In 3 cases aflR, ver 1 was not detected that HPLC method was approved too. Results shows PCR of critical gene of aflatoxin biosynthesis like aflR, ver 1 can be recommend as a good method for confirm of toxigenic aspergillus species. ITS sequencing showed that Aspergillus flavus Accession No MG430332.1 and Aspergillus parasiticus Accession No MH937579.1 are predominant in rice. It is recommended to study the species that produce ochratoxin (another important mycotoxin) in rice and other cereals.
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