Skip to main content
Log in

Practical method combining loop-mediated isothermal amplification and bait trap to detect Pythium helicoides from hydroponic culture solutions

Journal of General Plant Pathology Aims and scope Submit manuscript

Cite this article


Two detection methods combining loop-mediated isothermal amplification (LAMP) and a bait trap were developed to detect Pythium helicoides in greenhouses containing roses, miniature roses, and poinsettias in hydroponic culture systems. In “Bait-LAMP”, a crude extract derived from perilla seeds as the bait was used in the LAMP reaction, whereas in the “Bait culture-LAMP”, a crude extract of mycelia grown out from perilla seeds onto Pythium-selective medium served as the bait. The two methods are simple and rapid for practical monitoring of P. helicoides in hydroponic culture systems.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others


  • Cohen J (1960) A coefficient of agreement for nominal scales. Educ Psychol Meas 20:37–46

    Article  Google Scholar 

  • Feinstein AR, Cicchetti DV (1990) High agreement but low kappa: I. The problems with two paradoxes. J Clin Epidemiol 43:543–549

    Article  CAS  PubMed  Google Scholar 

  • Goto M, Honda E, Ogura A, Nomoto A, Hanaki K (2009) Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Biotechnology 46:167–172

    Article  CAS  Google Scholar 

  • Kageyama K (2011) Characteristics of high-temperature-growing Pythium species nowadays frequently occurring disease (in Japanese). Plant Prot 65:102–106

    Google Scholar 

  • Kageyama K (2014) Molecular taxonomy and its application to ecological studies of Pythium species. J Gen Plant Pathol 80:314–326

    Article  Google Scholar 

  • Kageyama K, Aoyagi T, Sunouchi R, Fukui H (2002) Root rot of miniature roses caused by Pythium helicoides. J Gen Plant Pathol 68:15–20

    Article  Google Scholar 

  • Ko WH, Chase LL, Kunimoto RK (1973) A microsyringe method for determining concentration of fungal propagules. Phytopathology 63:1206–1207

    Article  Google Scholar 

  • Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33:159–174

    Article  CAS  PubMed  Google Scholar 

  • McNemar Q (1947) Note on the sampling error of the difference between correlated proportions or percentages. Psychometrika 12:153–157

    Article  CAS  PubMed  Google Scholar 

  • Miyake N, Nagai H, Kageyama K (2014) Wilt and root rot of poinsettia caused by three high-temperature-tolerant Pythium species in ebb-and-flow irrigation systems. J Gen Plant Pathol 80:479–489

    Article  Google Scholar 

  • Mori Y, Nagamine K, Tomita N, Notomi T (2001) Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289:150–154

    Article  CAS  PubMed  Google Scholar 

  • Mori Y, Kitao M, Tomita N, Notomi T (2004) Real-time turbidimetry of LAMP reaction for quantifying template DNA. J Biochem Biophys Methods 59:145–157

    Article  CAS  PubMed  Google Scholar 

  • Morita Y, Tojo M (2007) Modifications of PARP medium using fluazinam, miconazole, and nystatin for detection of Pythium spp. in soil. Plant Dis 91:1591–1599

    Article  CAS  Google Scholar 

  • Nagamine K, Hase T, Notomi T (2002) Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 16:223–229

    Article  CAS  PubMed  Google Scholar 

  • Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:e63

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stanghellini ME, Rasmussen SL (1994) Hydroponics: a solution for zoosporic pathogens. Plant Dis 78:1129–1138

    Article  Google Scholar 

  • Takahashi R, Fukuta S, Kuroyanagi S, Miyake N, Nagai H, Kageyama K, Ishiguro Y (2014) Development and application of a loop-mediated isothermal amplification assay for rapid detection of Pythium helicoides. FEMS Microbiol Lett 355:28–35

    Article  CAS  PubMed  Google Scholar 

  • Thinggaard K, Middelboe AL (1989) Phytophthora and Pythium in pot cultures grown on ebb and flow bench with recirculating nutrient solution. J Phytopathol 125:343–352

    Article  Google Scholar 

  • van der Plaats-Niterink AJ (1981) Monograph of the genus Pythium. Stud Mycol 21:1–242

    Google Scholar 

  • Watanabe H, Kageyama K, Taguchi Y, Horinouchi H, Hyakumachi M (2008) Bait method to detect Pythium species that grow at high temperatures in hydroponic solutions. J Gen Plant Pathol 74:417–424

    Article  Google Scholar 

  • Waterhouse GM (1967) Key to Pythium pringsheim. Mycol Pap 109:1–15

    Google Scholar 

Download references


This work was supported in part by Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries and Food Industries from the Ministry of Agriculture, Forestry and Fisheries, Japan.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Noriyuki Miyake.

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Miyake, N., Nagai, H., Kato, S. et al. Practical method combining loop-mediated isothermal amplification and bait trap to detect Pythium helicoides from hydroponic culture solutions. J Gen Plant Pathol 83, 1–6 (2017).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: