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Nematode Extraction Methods: Root Extraction Techniques

Nematode Extraction Methods: Root Extraction Techniques. Kavitha Govindasamy Jimmy R.Rich Maria L .Mendes. A Plant-Parasitic Nematode. Sampling for Nematodes. Sampling and extracting for nematodes serves two purposes: Diagnosis a current problem Predict a future problem.

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Nematode Extraction Methods: Root Extraction Techniques

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  1. Nematode Extraction Methods:Root Extraction Techniques Kavitha Govindasamy Jimmy R.Rich Maria L .Mendes

  2. A Plant-Parasitic Nematode

  3. Sampling for Nematodes Sampling and extracting for nematodes serves two purposes: • Diagnosis a current problem • Predict a future problem

  4. Objectives of sampling and extracting nematodes Identify the nematode Population estimation Diagnose a nematode disease Make management decisions Regulatory purposes Research and surveys

  5. Some Common Nematode Extraction Methods • Root Samples Jar incubation and sieving Blending and sieving • Soil samples Baermann funnel Modified Baermann funnel • Foliar samples Punching and incubating Scissors technique

  6. Root Extraction Techniques:Jar Method Principle • Nematodes are extracted based on their active migration from roots

  7. Materials required • Glass jars and caps • 325 or finer mesh sieve • Labels and rubber bands • Dark chamber

  8. Sample ready for processing

  9. Wash the roots

  10. Damp dry the roots

  11. Weigh the damp-dried roots

  12. Place roots in jars, add water

  13. Place jars in tray

  14. Store in the dark for 72 hours

  15. Following incubation, remove from storage and add water

  16. Collect nematodes on sieve

  17. Rinse nematodes from sieve

  18. Nematode samples ready for observation

  19. Advantages and Limitations Advantages • Extracts migratory endoparasitic nematodes • Few materials needed • Simple procedure to follow Limitations • Not useful for ectoparasitic nematodes • Lack of aeration can be a problem • Sieves are needed to concentrate nematodes

  20. Root Extraction Techniques:Blending and Sieving Principle • Nematodes are released from roots due to maceration.

  21. Materials required • Blender • 40 and 325 or 500 mesh sieves • Pan or bowl • Wire mesh • Filter paper • Squeeze bottle • Weighing balance • Scissors

  22. Wash roots well

  23. Cut roots

  24. Damp dry the roots

  25. Weigh roots

  26. Place roots into blender, cover roots with water

  27. Blend at low speed for 10-20 seconds

  28. Pour the roots into 325 or finer mesh sieve

  29. Set up a Baermann tray

  30. Place filter paper on the Baermann tray

  31. Add water to Baermann tray

  32. Transfer the blended roots into the filter paper

  33. Incubate for 24 hours

  34. Remove roots from the pan

  35. Pour the water in the dish through a 325 or fine mesh sieve

  36. Concentrate nematodes for observation

  37. Advantages and Limitations Advantages • Fast method to extract nematodes from roots • Foliar nematodes can also be easily extracted Limitations • Chemicals released from the roots can be injurious to nematodes • Blending for long time may damage nematodes

  38. Acknowledgments • Janete Brito, Department of Plant Industry, Gainesville, Florida • Frank Woods, Nematode Assay Lab, University of Florida, Gainesville, Florida • Joey Orajay, Entomology and Nematology Department, University of Florida, Gainesville, FL

  39. For additional information or comments, please contact: • Kavitha_nem@yahoo.com • jimmyr@ufl.edu • mlmendes@ufl.edu

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