Cryopreservation of the Pinewood Nematode, Bursaphelenchus spp.
Abstract
Populations of three isolates of Bursaphelenchus xylophilus, the pinewood nematode, and one of B. mucronatus were treated with three cryoprotectants at -70 C for 24 hours followed by deep freezing at -180 C in liquid nitrogen for different periods of time. A solution of 15% glycerol, 35% buffer S, and 50% M9, or 1% aqueous solution of dimethylsulfoxide (DMSO), or a mixture of 60% M9 and 40% S buffer were used as cryoprotectants. A significantly larger number of juveniles than adults survived deep freezing. Significantly more nematodes were motile after cryopreservation in the 15% glycerol-S-M9 soludon than in the M9-S buffer solution or the DMSO aqueous solution. When cryopreserved nematodes that had been treated with glycerol solution were plated onto Botrytis cinerea, they reproduced rapidly over several generations. Cryopreserved nematodes were as pathogenic as untreated nematodes to Scots pines. Key words: Bursaphelenchus mucronatus, B. xylophilus, cryopreservation, cryoprotectant, nematode, pinewood nematode, Pinus sylvestris, Scots pine.Downloads
Published
Issue
Section
License
Copyright and Permissions
All material published by the Society of Nematologists (SON), except for papers prepared by United States and Canadian government employees, is copyrighted and protected under the U.S. copyright law. Under the Copyright Act of 1976, the term of copyright for materials registered by an organization is 75 years from the date first published. Before publishing any manuscript, SON requires that authors transfer full and complete ownership of any copyright to SON by signing a JON Page Charge/Copyright Form (.pdf). SON then registers the copyright. Subsequent use of published materials requires written permission from the SON and may be obtained by contacting the current Editor-in-Chief and state where and how the material will be used.
The author warrants that the article is an original work not published elsewhere in whole or in part, except in abstract form, and that the author has full power to make this grant. If portions of the article have been published previously, then the author warrants that permission has been obtained from the copyright holder and the author will submit a copy of the permission release with this copyright transfer form.
SON shall claim no proprietary right other than copyright. Authors and coauthors retain the right to revise, adapt, modify, or otherwise use all or part of the article in future works of the author(s), such as press releases, lectures, and reviews, provided that all such use is for the personal noncommercial benefit of the author(s). All patent rights are retained by the author(s).
