Journal of Korean Society of Forest Science (한국산림과학회지)

Korean Society of Forest Science (한국산림과학회)
권호 표지

Cryopreservation of Pine Wood Nematode, Bursaphelenchus xylophilus

  • Park, Ju-Yong (Division of Forest Diseases and Insect Pests, Korea Forest Research Institute) ;
  • Shin, Keum-Chul (Tree Pathology and Mycology Laboratory, Kangwon National University) ;
  • Kim, Kyung-Hee (Division of Forest Diseases and Insect Pests, Korea Forest Research Institute) ;
  • Lee, Seung Kyu (Division of Forest Diseases and Insect Pests, Korea Forest Research Institute) ;
  • Lee, Jong Kyu (Tree Pathology and Mycology Laboratory, Kangwon National University)
  • Received : 2005.08.23
  • Accepted : 2005.08.31
  • Published : 2005.10.31
Download PDF
⟨ Previous Next ⟩

Abstract

Pine wood nematode isolates from Haman, Gyoungsangnam-do were cryopreserved with three different cryoprotectants (glycerol, ethylene glycol and dimethylsulfoxide), Nematode Growth medium (NGM) and M 9 buffer solution, respectively, and then survival rate, reproduction ability, and pathogenicity of stored nematodes were compared. Survival rates of juvenile was excellent in 15% glycerol solution as $72{\pm}5.3%$. The survival ability of nematodes at juvenile stage was more efficient than that of nematodes at adult stage. Pre-incubation treatment at $4^{\circ}C$ before storing in liquid nitrogen didn't affect the survival ability of pine wood nematode. When the reproduction ability of nematodes was confirmed on Botrytis cinerea agar plates at $28^{\circ}C$ incubator, there was no difference between cryopreserved and non-cryopreserved. Pathogenic ability of cryopreserved nematodes was also evaluated and confirmed by artificially inoculating them on 2-year old red pine seedlings.

Keywords

References

  1. Bridge, J. and Ham, P.J. 1985. A technique for the cryopreservation of viable juveniles of Meloidogyne graminicola. Nematologica 31: 185-189 https://doi.org/10.1163/187529285X00247
  2. Chawla, M.L. and Prasad, S.K. 1975. Techniques in nematology. II. Comparative efficiency of sampling tools and nematode extraction methods. Indian Journal of Nematology 4: 115-123
  3. Curran, J., Gilbert, C. and Butler, K. 1992. Routine cryopreservafion of isolates of Steinernema and Heterorhabditis spp. Journal of Nematology 24:269-270
  4. Galway, N.J. and Curran, J. 1995. Cryopreservation of the Lesion Nematode (Pratylenchys spp.). Journal of Nematology 27: 483-485
  5. Ham, P.J., Townson, S., James, E.R. and Bianco, A.E. 1981. An improved technique for the cryopreservation of Onchocerca microfilariae. Parasitology 83: 139-146 https://doi.org/10.1017/S0031182000050113
  6. Hwang, S.W. 1970. Freezing and storage of nematodes in liquid nitrogen. Journal of Nematology 16:305-308 https://doi.org/10.1163/187529270X00324
  7. James, E.R. 1981. Schistosoma mansoni: Cryopreservation of Schistosomula by two-step addition of ethanediol and rapid cooling. Experimental Parasitology 52: 105-116 https://doi.org/10.1016/0014-4894(81)90066-7
  8. Luyet, B.J. and Keane, J.F. Jr. 1952. Comparative efficiency of ethylene glycol, glucose and sodium chloride in protecting tissues against freezing injury. Biodynamica 7: 119-131
  9. Moody, E.H., Lownsbery, B.E and Ahmed, J.M. 1973. Culture of the root-lesion nematode Pratylenchus vulnus on carrot disks. Journal of Nematology 5:225-226
  10. Popiel, I. and Vasquez, E.M. 1991. Cryopreservation of Steinernema carpocapase and bacteriophora. Journal of Nematology 23: 432-437
  11. Sayre, R.M. and Hwang, S.-W. 1985. Freezing and storing Ditylenchus dipsaci in liquid nitrogen. Journal of Nematology 7: 199-202
  12. Sulston, J. and Hodgkin, J. 1988. The nematode Caenorhabditis elegans. Methods. pp.587-605 in W. B. Wooel, ed. Cold Spring Harbor Laboratory, Cold Spring Harbor NY
  13. Triantaphyllou, A.C. and McCabe, E. 1989. Efficient preservation of root-knot and cyst nematodes in liquid nitrogen. Journal of Nematology 21: 423-426
  14. Viglierchio, D.R. and Schmit, R.Y. 1983. On the methodology of nematode extraction from field samples: Baermann funnel modifications. Journal of Nematology 15: 438-444