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Molecular characterisation of fungal culture collection by analysis of ITS1 sequences of rDNA

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Nevena Stević

on 18 April 2013

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Transcript of Molecular characterisation of fungal culture collection by analysis of ITS1 sequences of rDNA

Microbial culture collections and their significance Molecular characterisation of fungal culture collection
within genera Piptoporus, Pleurotus, Ganoderma and Schizophyllum
by analysis of ITS1 sequences of rDNA Providing the authenticated biological material, upon which high quality scientific research is based on
Deposits for key strains in science, industry and education

The aim of the culture collection is to maintain the purity, viability and genomic integrity of every single strain, being sources of key organisms for research, available for confirmation of results and further studies



Biological Resource Centers (BRCs) DNA barcoding of fungi Stević R. Nevena, Čapelja V. Eleonora, Galović O. Vladislava, Novaković S. Milana, Karaman A. Maja ITS region is an area of particular importance for molecular identification of fungi

Areas of high conservation and areas of high variability and presents an ideal starter for the development of specific PCR primers for identification of fungal species




Preferred DNA barcoding marker for the identification
of taxa and official primary barcoding marker for fungi The aim of this study was the molecularly based verification of morphological identification of lignicolous fungi obtained from the Culture Collection of Department of Biology and Ecology, Faculty of Sciences in Novi Sad Sequencing Visualisation and
extraction from gel F: ITS1
R: ITS4 Amplification of
ITS region Isolation of genomic DNA malt broth in 1.5 ml microtubes on room temperature with shaking for 3 days malt agar Petri plates at room temperature for
10-14 days Fungal cultivation Sequence analysis Alignment of the ITS sequences
using software MEGA 5.10

Comparison with sequences
from the NCBI database using BLAST Significant parameters used in database search Results and discussion MEGA 5.10 (Tamura et al, 2011) Calculation of homology within species or genus Alignment of sequences within species or genus Alignment of the ITS sequences using software MEGA 5.10 MEGA 5.10 (Tamura et al, 2011) Small interspecies variability of ITS region
Ideal marker for the species identification and verification of their integrity in cultures
Confirmation of morphological determination at the molecular level is the initial step in further research. Conclusion This work was supported in part by the Ministry of Education, Science and Technological Development of the Republic of Serbia within project No III43002 and by the Provincial Secretariat for Science and Technological Development, Vojvodina, Serbia ("Molecular and phenotypic diversity of taxa of economical and epidemiological importance, and endangered and endemic species in Europe").

Fungal cultures are obtained from the culture collection of Laboratory of Microbiology at the Department of Biology and Ecology, Faculty of Sciences in Novi Sad, while molecular analysis were performed at the Laboratory of Molecular Biology at the Institute of Lowland Forestry and Environment in Novi Sad. Acknowledgements Autochtonous, lignicolous species with significant biotechnological, pharmacological and industrial potential http://blast.ncbi.nlm.nih.gov/Blast.cgi from stock culture in distillated water
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