Molecular İdentification and Phylogeny of Some Hypocreales Members Isolated from Agricultural Soils

Year 2017, Volume: 8 Issue: 2, 137 - 142, 31.10.2017

Rasime Demirel

Abstract

Hypocreanlean fungi are include important plant pathogens around the
world. Head blight and crown rot disease of cereals caused by these species are
responsible for large economic losses due to reduction in seed quality and
contamination of grain with their mycotoxins. Although morphologic and
biochemical tests are still fundamental there is an increasing more towards
molecular diagnostics of these fungi. This paper reviews to PCR identification
of Hypocreanlean fungi isolated
from agricultural soil from Eskisehir. Five Hypocreanlean fungi belong to 4 different genera as Bionectria,
Fusarium, Gibberella and Nectria were isolated from 56 soil
samples. DNA of these strains
were isolated by glass beads and vortexing extraction method and used for PCR
amplification with universal fungal specific primers. The internal transcribed
spacer (ITS) regions of fungal ribosomal DNA (rDNA) were sequenced by CEQ 8000
Genetic Analysis System. The ITS-5.8S sequences obtained in this study were
compared with those deposited in the GenBank Database. Phylogenetic position of
investigated closely related Hypocreanlean fungi was determined.

Keywords

Hypocreanlean, PCR, ITS, Phylogeny, Eskişehir

Tarımsal Topraklardan İzole Edilen Bazı Hypocreales Üyelerinin Moleküler Teşhisi ve Filogenisi

Abstract

Hypocreanlean mantarları Dünya’daki önemli bitki patojenleridir. Bu türlerin sebep
olduğu baş tahribat ve tahıl hasar hastalığı, tohum kalitesinde azalma ve tahılın
mikotoksinler ile bulaşması nedeniyle büyük ekonomik kayıplardan sorumludur.
Morfolojik ve biyokimyasal testler hala temel olsa da, bu mantarların moleküler
teşhisine yönelim giderek artmaktadır. Bu makale, Eskişehir’deki tarım
topraklarından izole edilmiş olan Hypocreanlean mantarlarının PCR ile
identifikasyonunu incelemektedir. Bionectria, Fusarium, Gibberella ve Nectria
olmak üzere 4 farklı cinse ait 5 Hypocreanlean mantarı 56 farklı toprak
numunesinden izole edilmiştir. Bu suşların DNA'sı, cam boncuklar ve vorteks
ekstraksiyon yöntemi ile izole edilmiş ve üniversal fungal spesifik primerler
ile PCR amplifikasyonu için kullanılmıştır. Fungal ribozomal DNA'nın (rDNA) iç
transkripsiyonlu ayırıcı (ITS) bölgeleri, CEQ 8000 Genetik Analiz Sistemi ile
dizilenmiştir. Bu çalışmada elde edilen ITS-5.8S dizileri, GenBank veri tabanında
depolanan dizilerle karşılaştırılmıştır. İncelenen birbirine yakın akraba Hypocreanlean
mantarlarının filogenetik konumu belirlenmiştir.

Keywords

Hypocreanlean, PCR, ITS, Filogeni, Eskişehir

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