Identification of Aspergillus section Flavi Species from Peanuts and their Mycotoxigenic Properties

Yıl 2019, , 450 - 457, 31.12.2019

İşılay Lavkor

https://doi.org/10.24323/akademik-gida.667256

Öz

This study is aimed to identify 50 A. section Flavi isolates from peanut samples collected in the Adana and Osmaniye provinces of Turkey by using morphological and polymerase chain reaction (PCR), and to determine the in vitro aflatoxin and cyclopyazonic acid formation properties of the isolates. DNA was extracted for the identification of Aspergillus spp., and β-tubulin gene region on DNA was amplified by PCR. Identification of species was determined by comparing the result of sequences which belong to partial β-tubulin of the sequences with known ribosomal sequences using Basic Local Alignment Search Tool (BLAST). In the study, out of a total 50 fungal isolates, 17 were identified as A. parasiticus, and 33 were identified as A. flavus. As a result of Blast analysis of β-tubulin gene sequence comparisons A. flavus and A. parasiticus showed a major similarity in ranges 89-100% and 96-100%, respectively. Furthermore, all of the A. flavus and A. parasiticus isolates were determined as aflatoxin producers in the range from 0.45 to 4521.64 µg/L and 0.22 to 2562.87 µg/L with high-performance liquid chromatography (HPLC) under in vitro conditions, respectively. Total 32 A. flavus isolates produced cyclopiazonic acid in the range between 0.35-8.21 μg/g with thin layer chromatography (TLC). The present study provides a new method on molecular characterization of A. section Flavi on peanut samples collected from the Adana and Osmaniye provinces of Turkey. This research also demonstrated that A. flavus and A. parasiticus were responsible for mycotoxin contamination on peanut samples collected from Adana and Osmaniye provinces.

Anahtar Kelimeler

Aspergillus flavus, Aspergillus parasiticus, PCR, Mycotoxin, Peanut

Yerfıstığından İzole Edilen Aspergillus section Flavi Türlerinin Tanımlanması ve Mikotoksijenik Özelliklerinin Belirlenmesi

Öz

Bu çalışmada, Adana ve Osmaniye illerinden toplanan yerfıstığı örneklerinden izole edilmiş 50 Aspergillus section Flavi üyesi izolatın morfolojik ve polimeraz zincir reaksiyonu (PZR) yöntemiyle tanımlamaları yapılmıştır. Ayrıca, izolatların in vitro koşullarında aflatoksin ve siklopiazonik asit oluşturma özellikleri belirlenmiştir. Aspergillus spp. tanımlanmasında DNA ekstraksiyonları yapılmış, β-tubulin gen bölgesi ile PZR çoğaltılmıştır. Türlerin tanımlamasında β-tubulin gen bölgesine ait sekans sonuçları Blast (Basic Local Alignment Search Tool)’da bilinen ribozamal sekanslar ile karşılaştırılmıştır. Çalışmanın sonucunda, fungal izolatların 33’ü Aspergillus flavus, 17’si Aspergillus parasiticus olarak tanımlanmıştır. Blast analizi sonucunda ise A. flavus ve A. parasiticus türüne ait β-tubulin gen sekansı karşılaştırmaları sırası ile %89-100 ve %96-100 oranları arasında benzerlik göstermiştir. Ayrıca, A. flavus ve A. parasiticus izolatlarının tümü in vitro koşullarında yüksek performanslı sıvı kromatografisi (HPLC) ile sırasıyla 0.45 ile 4521.64 µg/L ve 0.22 ile 2562.87 µg/L aralığında aflatoksin üreticisi olduğu belirlenmiştir. İnce tabaka kromatografisiyle (TLC) 32 A flavus izolatı 0.35-8.21 μg/g aralığında siklopiazonik asit üretmiştir. Bu çalışma, Adana ve Osmaniye illerinden toplanan yerfıstığı örneklerinden izole edilen A. section Flavi'nin moleküler karakterizasyonu için yeni bir yöntem sunmaktadır. Ayrıca bu araştırma Adana ve Osmaniye illerinden toplanan yerfıstığı örneklerinden izole edilen A. flavus ve A. parasiticus izolatlarının mikotoksin oluşumundan sorumlu türler olduğunu göstermiştir.

Anahtar Kelimeler

Aspergillus flavus, Aspergillus parasiticus, PZR, Mikotoksin, Yerfıstığı

Kaynakça

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