American Journal of Microbiological Research, 2016, Vol. 4, No. 6, 178-180
Available online at http://pubs.sciepub.com/ajmr/4/6/4
©Science and Education Publishing
DOI:10.12691/ajmr-4-6-4
Identification of Nematode Trapping Fungus
Monacrosporium eudermatum Based on Genetic
Diversity Using RAPD Technique
Ali A. Kasim
*
, Maitham Dragh
Biological Department, College of Sciences, Misan University, Misan, Iraq
*Corresponding author: alimycol@yahoo.com
Abstract Nematode-trapping fungus Monacrosporium eudermatum a predacious fungus of nematodes, has been
very useful in understanding the most relationship between nematophagous fungi and their nematode hosts.
M.eudermatum is by far the common nematode-trapping fungus with the characteristic ability of forming adhesive
trapping nets once in contact with nematodes. Total DNA was extracted from M.eudermatum sera and amplified by
RAPD-PCR using three random primers (OPA-16, OPB-08, OPF-05). The results show amplification of all markers
(OPF-05, OPB-08, OPA-16) with the genomic DNA studied. As evidenced by the results, all the markers covered in
the study showed high polymorphism (up to 60%) with the exception of the primer (OPB-08) which showed no
polymorphism.
Keywords: nematophagous, Monacrosporium eudermatum, RAPD technique, PCR
Cite This Article: Ali A. Kasim, and Maitham Dragh, “Identification of Nematode Trapping Fungus
Monacrosporium eudermatum Based on Genetic Diversity Using RAPD Technique.” American Journal of
Microbiological Research, vol. 4, no. 6 (2016): 178-180. doi: 10.12691/ajmr-4-6-4.
1. Introduction
In recent years, different Polymerase Chain Reaction
(PCR) based molecular markers including Random
Amplified Polymorphic DNA (RAPD) have proved to be
useful tools for studying genetic diversity and monitoring
of soil borne fungi [1,2]. RAPD-PCR has the advantage of
being quick and easy and requires minute quantities of
genomic DNA for amplification. Furthermore, DNA
finger print can be generated with RAPD by using short
nucleotide (10-16 nucleotides bases) sequence of arbitrary
nature as primers and does not require any prior
knowledge of the target site sequence [3]. Several DNA-
based molecular markers such as; Randomly Amplified
Polymorphic DNA (RAPD), Restriction Fragment Length
Polymorphisms (RFLPs), Simple Sequence Repeats
(SSRs) and Amplified Fragment Length Polymorphism
(AFLP) have been successfully used to estimate genetic
diversity of fungi [4,5,6].
Nematophagous fungi are a diverse group of fungal
species that use refined mycelial structures or their conidia
to trap and capture nematodes and other small invertebrates.
This unique ability it made it possible to use these
fungi as agents of biological control against plant-and
animal-parasitic nematodes. However, effective application
of biological control factors in the field requires a
more information to understand the ecology and population
genetics of the nematophagous fungi in natural environments
[7].
Several studies using molecular markers to study the
diversity and evolution of nematode-trapping fungi.
Restriction fragment length polymorphism (RFLP) and
DNA sequencing of the internal transcribed spacer region
of the ribosomal RNA gene (ITS) region have found
genetic diversities among isolates within species in the
genera Arthrobotrys and Monacrosporium, showing cryptic
species among morphologically similar isolates [8,9,10].
Intraspecific differentiation derived from RFLP data and
polymorphisms in serine protease genes were also revealed
revealing among ecotypes of the nematophagous fungus
Pochonia chlamydospora based on host preferences [5,11].
The nematode-trapping fungus M.eudermatum belong
to the Orbiliomycetes, order Orbiliales and family
Orbiliaceae [12]. It is ubiquitous in a variety of habitats
such as a agricultural soils, farmed, fields, forests. The
present study was undertaken to determine if the three
markers (OPF-05, OPB-08 and OPA-16) amplify the genomic
DNA of the nematode-trapping fungus M.eudermatum
(monophyletic group Orbiliales) that isolated from farmed
soil of Misan governorate of Iraq.
2. Material and Methods
2.1. Isolates of M.eudermatum
Isolates of M.eudermatum were obtained from
microbiology laboratory at Biological Department, College
of Sciences, Misan University, Iraq and were isolated
from farmed soil of Misan, Governorate of Iraq).