Tech. D:3 



6. The blendor and sieving method developed by Taylor and Loegering 

 (1953) is essentially a means of freeing the nematodes from the 

 plant tissues so that they can be recovered by sieving. The 

 method is rapid because it does not depend upon the nematodes 

 moving out of the tissues by their own activity. The method 

 should be used with caution if infections are light, as some of 

 the specimens may be destroyed by the action of the blades or 

 may not be freed from the plant tissues. In practice, the plant 

 parts are cut into short pieces and placed in the blendor in a 

 small volume of water. The blendor is operated for about 10-20 

 seconds or until the plant pieces are reduced to small fragments. 

 The suspension is then washed simultaneously through a 2$ mesh 

 sieve to remove large debris particles, through a 6O mesh to 

 remove cyst-forming and female root-knot nematodes, and through 

 a 200 to 325 mesh sieve to remove other sizes and types of nema- 

 todes. 



This method is also useful for getting pieces of the cuticle of 

 female root-knot nematodes free from internal contents for exami- 

 nation and photomicrographs of peiuneal patterns. The residue 

 from the 60 mesh sieve is searched for these fragments which 

 will be numerous and recognizable if heavily infected roots have 

 been used. The differential stains previously mentioned for use 

 with the cyst-nematode recovery techniques may also have value 

 in this process, 



C. Staining in situ provides a means for detecting and studying nana- 

 todes within plant tissues without the necessity of making serially 

 sectioned slides v;ith the aid of a microtome. Plant parts treated 

 by the methods described may retain enough stain to be more or less 

 opaque if the tissues are too thick. The remedy for this is to 

 split the tissues. 



Several different methods for in situ staining of root and foliar 

 parts for nematodes are presented. 



