Çeşitli kaynaklardan izole edilen Aspergillus section Nigri türlerinin fenotipik ve moleküler biyolojik yöntemlerle karakterizasyonu
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Dosyalar
Tarih
2017
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Çalışmanın amacı, Aspergillus section Nigri grubu içerisinde yer alan
mikrofungusların tür düzeyinde morfolojik ve moleküler biyolojik özelliklerine
dayalı olarak karakterize edilmesidir. Bu amaç için, İzmir ili ve çevresinden çeşitli
hava, toprak ve gıda örnekleri alınarak, bu örneklerde bulunan section Nigri üyesi
Aspergillus kolonileri izole edilmiştir.
İzole edilen mikrofungus türlerinin tanısı Klich 2002’de belirtildiği üzere 5
farklı besiyerinde makro ve mikromorfolojik özelliklerin tayini ile fenotipik
olarak yapılmıştır.
Moleküler tanılama amacıyla DNA izolasyonunda UltraClean® Microbial
DNA Isolation Kiti (MoBio Laboratories, Inc.), EZNA Fungal DNA Kiti (Omega
Bio-tek., Inc.) kullanılmıştır. Tüm Aspergillus section Nigri türlerinin kalmodulin
sekans verisi ile birbirinden ayırt edilebilmesi sebebiyle Varga et al 2007’de
belirtilen şekilde kalmodulin geninin filogenetik analizi gerçekleştirilmiştir.
Çalışmanın sonucunda gıda ürünleri, toprak ve hava gibi çeşitli kaynaklardan
toplam 150 adet Aspergillus section Nigri üyesi izole edilmiştir. Morfolojik
çalışmalar sonucunda 5 farklı tür (A. awamori, A. carbonarius, A. foetidus, A.
japonicus ve A. niger) elde edilirken moleküler çalışmalar sonucunda 10 farklı tür
(A. acidus, A.acuelatus, A. awamori, A. carbonarius, A. foetidus, A. japonicus, A.
neoniger, A. niger, A. tubingensis, A. uvarum) tanılanmıştır.
Elde edilen moleküler veriler fenotipik tanıyla tanılamayan 5 türün varlığını
göstermiştir. Böylelikle çalışma sonucunda özellikle Aspergillus section Nigri
üyeleri olmak üzere mikrofungusların tanısında sadece fenotipik yöntemlerinin kullanımının subjektif gözlemlere dayalı olması, tanı anahtarlarının güncel
olmaması nedeniyle yetersiz olduğu açığa konmuştur.
Aspergillus section Nigri which is composed of microfungi that possess importance in food mycology, biotechnology and industrial microbiology is one of the most complicated and complex groups in mycology. In order to facilitate the identification of this taxonomically complicated group, isolation from different sources such as soil, air and food and identification in species level by means of polyphasic taxonomy using morphological, physiological and molecular biological methods is aimed by comparing conventional phenotypical identification and molecular identification methods in this study. Black spored Aspergillus strains isolated from various food products such as dried fruits, muesli, breakfast cereals sold in Izmir district, air and soil were firstly morphologically identified in genus level and species level according to Barnett (1999) and Klich (2002) and Samson (2010) respectively. UltraClean® Microbial DNA Isolation Kit (MoBio Laboratories, Inc.), EZNA Fungal DNA Kit (Omega Bio-tek., Inc.) and a manual method were used for DNA isolation. Since all of the species of Aspergillus section Nigri can be distinguished with kalmodulin sequence data, phylogenetical analysis of this gene was performed. 150 isolates of Aspergillus section Nigri were isolated from several sources such as foodstuff, air and soil. While 5 different species (A. awamori, A. carbonarius, A. foetidus, A. japonicus and A. niger) were identified according to morphological identifications, molecular identifications revealed 10 different species (A. acidus, A.acuelatus, A. awamori, A. carbonarius, A. foetidus, A. japonicus, A. neoniger, A. niger, A. tubingensis, A. uvarum). Molecular data showed that species that there are species that can not be identified solely by phenotypical identification. And it is shown that relying on phenotypical methods is inadequate since it depends on subjective observations and keys used for identifications may not be not up-to-date.
Aspergillus section Nigri which is composed of microfungi that possess importance in food mycology, biotechnology and industrial microbiology is one of the most complicated and complex groups in mycology. In order to facilitate the identification of this taxonomically complicated group, isolation from different sources such as soil, air and food and identification in species level by means of polyphasic taxonomy using morphological, physiological and molecular biological methods is aimed by comparing conventional phenotypical identification and molecular identification methods in this study. Black spored Aspergillus strains isolated from various food products such as dried fruits, muesli, breakfast cereals sold in Izmir district, air and soil were firstly morphologically identified in genus level and species level according to Barnett (1999) and Klich (2002) and Samson (2010) respectively. UltraClean® Microbial DNA Isolation Kit (MoBio Laboratories, Inc.), EZNA Fungal DNA Kit (Omega Bio-tek., Inc.) and a manual method were used for DNA isolation. Since all of the species of Aspergillus section Nigri can be distinguished with kalmodulin sequence data, phylogenetical analysis of this gene was performed. 150 isolates of Aspergillus section Nigri were isolated from several sources such as foodstuff, air and soil. While 5 different species (A. awamori, A. carbonarius, A. foetidus, A. japonicus and A. niger) were identified according to morphological identifications, molecular identifications revealed 10 different species (A. acidus, A.acuelatus, A. awamori, A. carbonarius, A. foetidus, A. japonicus, A. neoniger, A. niger, A. tubingensis, A. uvarum). Molecular data showed that species that there are species that can not be identified solely by phenotypical identification. And it is shown that relying on phenotypical methods is inadequate since it depends on subjective observations and keys used for identifications may not be not up-to-date.
Açıklama
Anahtar Kelimeler
Aspergillus Section Nigri, İzolasyon, Tanılama, Polifazik Taksonomi, Moleküler Tiplendirme, PCR, Isolation, Identification, Polyphasic Taxonomy, Molecular Typing
