SylbV01-00-00. pmd

Unit VI
Application of molecular techniques in Nematology.
VI. Practical
• Collection of soil and plant samples;
• Extraction of nematodes from soil by Baermann funnel, sieving and decanting,
elutriation and sugar centrifugal methods;
• Extraction of cysts from soil;
• Extraction of nematodes from plant material;
• Estimation of population densities;
• Staining plant material for nematodes;
• Killing and fixing nematodes, clearing nematodes by slow and Seinhorst’s methods;
• Preparation of temporary and permanent mounts;
• Measurements, drawing, microphotography, special preparation of nematodes –
perineal patterns, vulval cones, en-face and body sections;
• Collection of root exudates, preparation of exhibits of nematode diseased plant
material, 
in-vitro 
culturing techniques of nematodes- callous culture, excised root
and carrot disc techniques.
VII. Suggested Reading
Ayoub SM. 1981. 
Plant Nematology – An Agricultural Training Aid
.
Barker KR, Carter CC and Sasser JN. 1985. 
An Advanced Treatise on 
Meloidogyne. Vol. II.
Methodology
. International 
Meloidogyne 
Project, NCSU, Raleigh. USA.
Manzanilla-Loìpez, RH and Marbaìn-Mendoza N. 2012. 
Practical Plant Nematology
, Montecillo,
Texcoco: Biblioteca Basica de Agricultura.
Sikora RA, Coyne D, Hallman J and Timper P. 2018. 
Plant Parasitic Nematodes in Subtropical
and Tropical Agriculture.
3rd edn
. 
CABI Publishing, England.
Southey JF. 1986. 
Laboratory Methods for Work with Plant and Soil Nematodes
. HMSO, London.
Subbotin SA, Mundo-Ocampo M and Baldwin J. 2010. 
Systematics of The Genus Heterodera in
Systematics of Cyst Nematodes
(Nematoda: Heteroderinae), Part B, Series: Nematology
Monographs and Perspectives, Volume: 8B, Brill.
Zuckerman BM, Mai WF and Harrison MB. 1985. 
Plant Nematology Laboratory Manual
. Univ.
of Massachusetts.