Chapter 17
Nematoda
Although the class Nematoda consists of many species ranging in size
from the large Ascaris lumbricoides to the small plant parasitic nematodes, only a few attempts have been made to culture the cells or tissues
of nematodes. However, a continuous cell line has been reported from
the plant parasitic nematode Meloidogyne incognita (Manousis and Ellar,
1990).
Subprotocol 1
Whole Bodies of Plant Parasitic Nematodes (Tylenchida)
The following is based on the methods of Manousis and Ellar (1990).
Materials
- Centrifuge and centrifuge tube
- Forceps
- Culture tube
- 15-cm 2 plastic culture bottle
- 0.525% Sodium hypochlorite
- 0.9% NaCl
- Leibovitz's L-15 medium
- Schneider's Drosophila medium (see Appendix 1)
- The root knot nematode M. incognita, reared on tomato roots
Procedure
PROTOCOL
Equipment
Solutions
Animals
1. Remove mature egg-laying females (Fig.l) from infected tomato roots Sterilization
using fine forceps.
2. Place the nematodes in 0.9% NaCI in a culture tube.
3. When enough individual nematodes (approximately 15) have been
accumulated, remove excess solution.
4. Surface sterilize the nematodes in a solution of 0.525% sodium hypochlorite for 6 min.
5. Transfer the nematodes to a sterile tube.
6. Wash the nematodes twice under aseptic conditions with a large volume of sterile tap water.