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
2015
Kawanobe, M. ((Tokyo University of Agriculture and Technology, Koganei, Tokyo (Japan). Graduate School of Bio-Applications and Systems Engineering), (Agri-RAND, Bunkyo-ku, Tokyo (Japan))) | Miyamaru, N. | Yoshida, K. | Kawanaka, T. | Toyota, K.
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 = log sub(2) (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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