Characterization of Cladosporium Species by Internal Transcribed Spacer-PCR and Microsatellites-PCR

Characterization of Cladosporium Species by Internal Transcribed Spacer-PCR and Microsatellites-PCR

This investigation compared genetic similarities and diversities within and among Cladosporium species populations using the two PCR-based markers; Internal Transcribed Spacer (ITS)-PCR and microsatellite-PCR. Nuclear ribosomal DNA internal transcribed spacers have been used to analyze intraspecific...

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Bibliographic Details
Published in:Pakistan journal of biological sciences Vol. 19; no. 4; pp. 143 - 157
Main Authors: Alhussaini, Mohammed S, Adbo Moslem, Mohamed, Alghonaim, Mohammed I, Al-Ghanayem, Abdullah A, Al-Yahya, Abdulrahman A I, Hefny, Hamido M, Saadabi, Adbul Moneam
Format: Journal Article
Language:English
Published: Pakistan 2016
Subjects:
Cladosporium
Cladosporium - classification
Cladosporium - genetics
Cluster Analysis
DNA Fingerprinting - methods
DNA Primers
DNA, Fungal - genetics
DNA, Ribosomal Spacer - genetics
Genetic Variation
Genotype
Microsatellite Repeats
Phylogeny
Polymerase Chain Reaction - methods
Cladosporium
microsatellite
MspI
DNA fingerprinting
CfoI
RsaI
ITS-PCR
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Summary:This investigation compared genetic similarities and diversities within and among Cladosporium species populations using the two PCR-based markers; Internal Transcribed Spacer (ITS)-PCR and microsatellite-PCR. Nuclear ribosomal DNA internal transcribed spacers have been used to analyze intraspecific and interspecific relationships in various fungi. In the present study, the internal transcribed spacer (ITS)-PCR and microsatellite-PCR were used to identify the genetic diversities in Cladosporium species. The Internal Transcribed Spacer (ITS) was amplified using polymerase chain reaction combining primers ITS4 and ITS5. The PCR products were digested with three restriction enzymes and separated by agarose gel electrophoresis. Restriction patterns generated by CfoI and Msp I and RsaI were unique for most species assayed. The ITS-PCR fingerprinting methods led to a clear differentiation of the isolates at the species level. Fingerprinting profiles generated readily discriminated between each of the 6 species. Cluster analysis further supported this observation and clusters corresponding to each species were readily identified in the dendrograms. Seven microsatellite primers out of eight primers were unable to generate visible DNA fingerprints. Amplification experiments demonstrated that microsatellite primer, T3B and (GTG) 5 are technically simple tools for assaying genetic variability in Cladosporium spp.
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ISSN:1028-8880
1812-5735
DOI:10.3923/pjbs.2016.143.157