Manitoba Survey and Molecular Quantification of Soybean Cyst Nematode

Crop Soybean
Start Date2017
End Date2019
Principal InvestigatorTenuta, Mario, University of Manitoba
MPSG Financial Support$243,225
Total Project Funding$243,225
ReportTenuta – SCN Survey – ANNUAL report

Research Objectives

  1. Survey soybean fields for SCN
  2. Publish results of this and the past two SCN surveys
  3. Develop accurate and quick molecular method for quantification of SCN in soil

Project Description

This project continues the proactive role of the Univerity of Manitoba and MPSG in knowing when SCN has arrived in the Province, outreach activities to inform growers about SCN and develop capacity within the Province for future surveys and research to mitigate damage due to SCN after its arrival. 25 soybean fields will be surveyed. In addition, fields of a seed potato grower in North Cypress will sampled. Each field will be sectioned into areas prone for the establishment of SCN. Areas will be sampled to 12” with 10 to 25 sub-samples taken for each, depending on section area. Soil from each area will be air-dried and 2.5 lbs will be extracted using a cyst extractor. Extracted cysts will be isolated from soil debris by flotation in ethanol. They will then be screened for belonging to the genus Heterodera based on vulval cone structure and being lemon-shaped cysts. The cysts of Heterodera will then be sorted if being SCN using identification keys. Suspect SCN cysts will then be confirmed using PCR methods. Only samples being positive by morphological and molecular examination will be considered to be SCN. Positive field will be resampled.
We propose to develop an acurate and quick molecular method by skipping the most laborious steps (crushing and counting) and then manual SCN identification. Instead we propose a genonomic DNA extraction of isolated cysts with quantification using the CoxII gene and real time PCR. Twenty soils from Ontario ranging from none to extremely high cyst levels will be used to optimize a cyst extraction and PCR protocol as well as calibrate tthe real time PCR method.