Nov 20, 2020

Public workspaceFungal Extraction from Beetle Galleries

DOI
dx.doi.org/10.17504/protocols.io.bnutmewn
  • You Li1,
  • Jiri Hulcr1
  • 1University of Florida
  • Protocols Bark Beetle Mycobiome
Bark Beetle Mycobiome Research Coordination Network Bark
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DOI: dx.doi.org/10.17504/protocols.io.bnutmewn
Document Citation: You Li, Jiri Hulcr 2020. Fungal Extraction from Beetle Galleries. protocols.io https://dx.doi.org/10.17504/protocols.io.bnutmewn
License: This is an open access document distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: October 23, 2020
Last Modified: November 20, 2020
Document Integer ID: 43635
Abstract
This protocol describes how to extract fungi from beetle galleries.

This protocol is part of the Bark Beetle Mycobiome (BBM) Research Coordination Network. For more information on the BBM international network: Hulcr J, Barnes I, De Beer ZW, Duong TA, Gazis R, Johnson AJ, Jusino MA, Kasson MT, Li Y, Lynch S, Mayers C, Musvuugwa T, Roets F, Seltmann KC, Six D, Vanderpool D, & Villari C. 2020. Bark beetle mycobiome: collaboratively defined research priorities on a widespread insect-fungus symbiosis. Symbiosis 81: 101–113 https://doi.org/10.1007/s13199-020-00686-9.

Materials


(all autoclaved)
  • eppendorf vials
  • Pestles or some other grinding tool
  • scalpel
  • Reserve hood time
  • strong forceps
  • vials
  • ethanol in petri dish for dipping the forceps & scalpel into
  • vial rack
  • (chisel)
  • (pruner)
  • pipette
  • 1000 uL pipette tips
  • EtOH bath

For field sampling of beetle associated fungi:
  • ethanol
  • squirt bottle (or separate vials with ethanol)
  • hard forceps
  • scalpel
  • scalpel blades
  • lighter (for forceps & scalpel sterilization)
  • ziploc bags
  • permanent markers (alcohol-resistant) 


Procedure

Scrape piece of 1 gallery in the hood, scrape into
a. 1 ml of 1X PBS AUTOCLAVED phosphate buffer saline solution in tube
3. mortar & pestle
4. plate directly, refrigeration decreases growing success
5. Plate three different dilutions, refer to the dilution protocol.