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Molecular diagnostic key for identification of single juveniles of seven common and economically important species of root‐knot nematode ( Meloidogyne spp.)
Author(s) -
Adam M. A. M.,
Phillips M. S.,
Blok V. C.
Publication year - 2007
Publication title -
plant pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.928
H-Index - 85
eISSN - 1365-3059
pISSN - 0032-0862
DOI - 10.1111/j.1365-3059.2006.01455.x
Subject(s) - biology , rapd , primer (cosmetics) , polymerase chain reaction , terra incognita , nematode , root knot nematode , genetics , dna sequencing , ribosomal dna , dna , botany , meloidogyne incognita , ecology , gene , phylogenetics , genetic diversity , population , chemistry , demography , organic chemistry , sociology
A molecular protocol is presented for distinguishing seven of the most common and economically important Meloidogyne spp. DNA was extracted from individual second‐stage juvenile (J2) nematodes of Meloidogyne spp. and amplified by PCR (polymerase chain reaction). Fifteen PCRs including amplification of rDNA, specific SCAR (sequence characterized amplified region) and RAPD (random amplified polymorphic DNA) fragments were possible from the extracted DNA. This enabled a molecular diagnostic key for M. incognita , M. javanica , M. arenaria , M. mayaguensis , M. hapla , M. chitwoodi and M. fallax to be designed. The key unifies published methods into a single logical schematic using primer combinations that were previously validated and shown to work reliably and specifically. The protocol can be used with single juvenile or adult nematodes and the schematic can readily be expanded to accommodate more species. The use of RAPD amplification to assist with identification of samples which do not yield diagnostic amplification products after the first three steps of the molecular key is also described.