Abstract
Filamentous fungi were collected from diseased hop (Humulus lupulus; L.) and DNA was prepared from 19 isolates. The critical step of cell lysis was carried out using zymolyase for fungal cell wall digestion. DNA was successfully prepared for all isolates and allowed ribosomal DNA region amplifications by PCR. The amplicons were sequenced and sequences were compared with nucleotides databases. These analyses allowed identification of each fungus and gave a precise view of the ecosystem. Seven different genera were found among the 19 isolates. A phylogenetic tree was constructed and revealed the diversity of the hop plant environment.
Similar content being viewed by others
References
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389–3402.
Bonants P, Hagenaar-de Weerdt M, van Gent-Pelzer M, Lacourt I, Cooke D, Duncan J (1997) Detection and identification of Phytophthora fragariae Hickman by the polymerase chain reaction. Eur. J. Plant Pathol. 103: 345–3556.
Do N, Adams RP (1991) A simple technique for removing plant polysaccharide contaminants from DNA. BioTechniques 10: 162–166.
Griffin DW, Kellogg CA, Peak KK, Shinn EA (2002) A rapid and efficient assay for extracting DNA from fungi. Lett. Appl. Microbiol. 34: 1–5.
Hatsch D, Phalip V, Jeltsch J-M (2002) Development of a bipartite method for Fusarium identification based on cellobiohydrolase-C: CAPS and Western blot analysis. FEMS Microbiol. Lett. 213: 245–249.
Raina K, Chandlee JM (1996) Recovery of genomic DNA from a fungus (Sclerotinia homoeocarpa) with high polysaccharide content. BioTechniques 21: 1030–1032.
Smith GW, Ives LD, Nagelkerken IA, Ritchie KB (1996) Caribbean sea-fan mortalities. Nature 383: 487.
Van Burik J-A H, Schreckhise RW, White TC, Bowden RA (1998) Comparison of six extraction techniques for isolation of DNA from filamentous fungi. Med. Mycol. 36: 299–303.
Wach A, Pick H, Philippsen P (1994) Procedures for isolation of yeast DNA for different purposes. In: Jonhston JR, ed. Molecular Genetics of Yeast. A Practical Approach. Oxford: IRL Press, Oxford University Press, pp. 1–16.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Phalip, V., Hatsch, D. & Jeltsch, JM. Application of a yeast method for DNA extraction associated with database interrogations for the characterization of various filamentous fungi from diseased hop. Biotechnology Letters 26, 409–413 (2004). https://doi.org/10.1023/B:BILE.0000018260.26171.6f
Issue Date:
DOI: https://doi.org/10.1023/B:BILE.0000018260.26171.6f