Feb 28, 2024

Public workspaceSCN Cyst Extraction from Greenhouse Cultures (crocks)

DOI
https://dx.doi.org/10.17504/protocols.io.q26g7pod8gwz/v1
  • Casey Schlenker1
  • 1The Ohio State University
  • OSU Soybean Pathology and Nematology
Casey A Schlenker
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DOI: https://dx.doi.org/10.17504/protocols.io.q26g7pod8gwz/v1
Protocol Citation: Casey Schlenker 2024. SCN Cyst Extraction from Greenhouse Cultures (crocks). protocols.io https://dx.doi.org/10.17504/protocols.io.q26g7pod8gwz/v1
License: This is an open access protocol 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
Protocol status: Working
We use this protocol and it's working
Created: February 28, 2024
Last Modified: February 28, 2024
Protocol Integer ID: 95907
Keywords: scn cyst extraction from greenhouse culture, scn cyst extraction, greenhouse culture, extraction
Abstract
SCN Cyst Extraction from Greenhouse Cultures (crocks)
Materials
Preparing Green House Cultures for Cyst Extraction
  1. Gardening shears
  2. Cart
  3. Gloves
  4. 3-gallon bucket

SCN cyst extraction
  1. Petri Dishes, small, 60x15mm 
  2. Trays 
  3. 20 and 60 Mesh Sieves  
  4. 3 Gallon Waste Bucket 
  5. Gray Bucket 
  6. White Plastic Cutting Board with Hole 
  7. Wash Bottle, Wide Nozzle 
  8. Plastic Dish Apron 
  9. Gloves 

Cleaning the crocks
  1. Gloves 
  2. Scrub pad 
  3. Pipe Cleaner 
  4. Bleach 

Preparing Green House Cultures for Cyst Extraction
Put on gloves to protect hands before beginning preparation
Using gardening shears, clip off all of the stems on the chosen soybean plant, leaving behind 1-2 inches of the stem. Place stems in the 3-gallon bucket.
Place crocks on cart for transport back to the lab. Dispose of the soybean clippings in either the steam cart or the compost.
SCN cyst extraction
*Once you begin this process for a crock you must finish the whole crock within the work day. 
Lay out small petri dishes on a tray and set aside.
To begin collecting cysts from greenhouse cultures, flood the crock gently with water. Utilize the faucet with long rubber tubing.
A white cutting board with a hole is placed on-top of a white tub. On top of the cutting board hole place the 20/60 mesh sieves.
Fill a grey bucket with water and place close to crock.
Once properly soaked, remove the long skinny tube from the center of the crock, and place aside.
Choose your first sample. Remove the tag from the sample and set aside. Pull the stem and the tube out of the crock at the same time, being careful not to rip the stem.
Move the root and tube to the gray wash bucket and gently swirl to dislodge any sand and turface from the tube.
Place the washed root on top of the 20/60 sieves and blast the roots, with the handheld sprayer, turning the root over periodically to ensure all cysts are properly dislodged. A good indicator of when all the cysts are removed is when all the turface is dislodged from the roots.
After checking that the root is thoroughly clear of cysts, dispose of the root in a 3-gallon bucket and rinse 20 sieve briefly.
Using a wash bottle, wash all material from 60 sieve into a small petri dish. Replace the petri dish lid and place the tag from the corresponding sample on top of the petri dish.
Repeat steps 6-10 with the remaining tubes.
Cleaning the crocks
To clean the crocks, you must thoroughly wash all growing substrate and plant matter out of the crock. Scrub the crocks to ensure all previous plant material is removed.
Place all the pvc tubing into the crock (22 in total, 1 long tube) and fill 80% with water. Fill the rest of the crock with bleach. Let sit for a minimum of 24 hours.
Drain and let dry.