Nematological Research (Japanese Journal of Nematology)
Online ISSN : 1882-3408
Print ISSN : 0919-6765
ISSN-L : 0919-6765
ORIGINAL ARTICLES
Quantification of lesion nematode (Pratylenchus zeae), stunt nematode (Tylenchorhynchus leviterminalis), spiral nematode (Helicotylenchus dihystera), and lance nematode (Hoplolaimus columbus), parasites of sugarcane in Kitadaito, Okinawa, Japan, using real-time PCR
Masanori KawanobeNaoko MiyamaruKoichi YoshidaTakeshi KawanakaKoki Toyota
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2015 Volume 45 Issue 1 Pages 35-44

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Abstract

The lesion nematode (Pratylenchus zeae), the stunt nematode (Tylenchorhynchus leviterminalis), the spiral nematode (Helicotylenchus dihystera), and the lance nematode (Hoplolaimus columbus) were major plant-parasitic nematode (PPN) species identified in the sugarcane fields on Kitadaito Island, Okinawa, Japan, for which real-time PCR primer sets were developed. Since the four PPN species ubiquitously inhabited Kitadaito, P. penetrans, which was confirmed to be absent in Kitadaito, was used as a benchmark to establish a soil calibration curve. Kitadaito soil samples were inoculated with juveniles and adults of P. penetrans at densities of 8, 32, 128, and 256/10 g fresh soil, and a soil calibration curve was obtained: y =-0.95x + 34.83 (y = Ct values, x = log2 (the number of P. penetrans ). The soil calibration curve was consistent with a calibration curve derived from serially diluted DNA samples of handpicked single P. penetrans (y =-1.01x + 34.73), after adjusting the DNA dilution rates to the densities in soil. The results suggested that calibration curves developed using serially diluted DNA samples (y =-1.13x + 37.00, y =-0.99x + 30.76, y =-1.06x + 35.52, and y =-1.00x + 28.96 for P. zeae, T. leviterminalis, H. dihystera, and H. columbus, respectively) may be appropriate to quantify target PPN in Kitadaito soil.

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© 2015 The Japanese Nematological Society
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