Response to Bortolotti et al. 2012-a re-evaluation of our polymerase chain reaction-restriction fragment length polymorphism genotyping method

2013 ◽  
pp. n/a-n/a
Author(s):  
F. M. B. Zambra ◽  
J. A. B. Chies ◽  
C. S. Alho ◽  
T. D. Veit
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Genotyping Method ◽  
Polymerase Chain ◽  
Restriction Fragment Length

scholarly journals Distinguishing Heterodera filipjevi and H. avenae Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism and Cyst Morphology

2010 ◽  
Vol 100 (3) ◽  
pp. 216-224 ◽  
Author(s):  
Guiping Yan ◽  
Richard W. Smiley
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Cyst Nematodes ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Restriction Fragment Length

The cereal cyst nematodes Heterodera filipjevi and H. avenae impede wheat production in the Pacific Northwest (PNW). Accurate identification of cyst nematode species and awareness of high population density in affected fields are essential for designing effective control measures. Morphological methods for differentiating these species are laborious. These species were differentiated using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) of internal transcribed spacer (ITS)-ribosomal (r)DNA with up to six restriction endonucleases (TaqI, HinfI, PstI, HaeIII, RsaI, and AluI). The method was validated by inspecting underbridge structures of cyst vulval cones. Grid soil sampling of an Oregon field infested by both species revealed that H. filipjevi was present at most of the infested grid sites but mixtures of H. avenae and H. filipjevi also occurred. These procedures also detected and differentiated H. filipjevi and H. avenae in soil samples from nearby fields in Oregon and H. avenae in samples from Idaho and Washington. Intraspecific polymorphism was not observed within H. filipjevi or PNW H. avenae populations based on the ITS-rDNA. However, intraspecific variation was observed between H. avenae populations occurring in the PNW and France. Methods described here will improve detection and identification efficiencies for cereal cyst nematodes in wheat fields.

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