Determining fungal diversity on Dendroctonus ponderosae and Ips pini affecting lodgepole pine using cultural and molecular methods

Woon Lim Young, Jae Jin Kim, Monica Lu, Colette Breuil

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Several beetles (Coleoptera: Scolytidae) and their fungal associates cause severe damage to lodgepole pine in Western Canada and the Northwestern United States. The fungal diversity from the surface of two bark beetle species, Dendroctonus ponderosae Hopkins (mountain pine beetle) and Ips pini Say (pine engraver), was surveyed using cultural and molecular methods. Nine fungal taxa were recognised by morphological characterizations. All nine taxa were isolated from the mountain pine beetle whereas only seven of the nine taxa were isolated from the pine engraver. The identification was based on cultural morphology and high sequence similarities of the internal transcribed spacer (ITS) and large subunit ribosomal DNA (LSU rDNA) region to sequences of known fungi. Fungal ITS regions were amplified from DNA directly extracted from the beetle surface. The PCR products were cloned and 250 clones were classified by their restriction pattern with HaeIII and RsaI. A total of 26 RFLP types were identified and subsequently sequenced. Among them, 15 RFLP types were identified as being present in mountain pine beetle and 14 were present in pine engraver. Sequence analysis of the RFLP types showed that 23 ascomycetes and 3 basidiomycetes were represented in the clone libraries, whereas the isolates from the cultural method represented 7 ascomycetes and 2 basidiomycetes. We found that yeast and non-staining filamentous Euascomycetes fungi were detected efficiently using a molecular approach, while the major sapstaining fungi and decay fungi were best detected using cultural methods.

Original languageEnglish
Pages (from-to)79-94
Number of pages16
JournalFungal Diversity
Volume19
Publication statusPublished - 2005 Jun 1
Externally publishedYes

Keywords

  • Bark beetle
  • Fungal diversity
  • ITS, lodgepole pine, mountain pine beetle

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Ecology

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