Canadian Agricultural Adaptation Program (CAAP)
Final Report
Applicant Name:
Project Title:
MRAC Program Coordinator:
Project Number:
Reporting Period:
Date of Submission:
Manitoba Pulse Growers Association
Survey for Soybean Cyst Nematode (SCN) and Development of
Diagnostic Capacity in Manitoba
Kristin Yaworski-Lowdon
CAAP MB0436
Final Report
October 10, 2013
Executive Summary
1. Briefly summarize the activities funded in this project, highlighting key achievements and results. Your
summary should provide answers to the questions below.
(a) What was the purpose of the project?
(b) Why are the project and its results significant for the target group and/or stakeholders?
(c) What are the actual achievements/outcomes, and any unintended impacts?
(d) Were there any aspects not completed?
(e) What were the lessons learned?
(f) What factors contributed to the project’s success or non-success?
(g) What are the next steps? (Not all projects will have next steps, but often there are activities planned for
after the end of the project that are directly or indirectly a result of the project.)
(h) Is the solution or strategy likely to be further implemented or is the innovative product, process or
technology likely to be adopted by the sector?

If you answered yes to the above, describe what you expect will be the next activities.

If you answered no, explain why not.
This report details results of soil sampling and analyses targeted for Heterodera glycines, (Soybean Cyst
Nematode; SCN), conducted in 2012-2013, in Province of Manitoba, Canada. Heterodera glycines Ichinohe,
1952, is recognized as the major pest of soybean (Glycine max) worldwide. Soybean is the only major
economically important host plant for this nematode. Estimated loss of 7% in the United States each year are
due to this pest with damage costing up to $1.5 billion per year (Wrather et al. 2001, Benjamin et al. 2004).
With respect to the importance of soybean cultivation for Canadian farmers, early detection and precise
identification is of significant important. SCN has rapidly moved northward in the mid US states. It is now
present in some of the counties Manitoba borders with North Dakota and Minnesota. It is only time until it is
Manitoba, if not already. Recently, the Canadian Food Inspection Agency has declassified SCN as a regulated
pest in Canada. This means to farmers that surveys for the nematode are not to be done in the future by the
agency.
Thus the current project was initiated between the University of Manitoba, the Manitoba Pulse Growers
Association and Manitoba Agriculture, Food and Rural Affairs to survey for the presence of SCN in
Manitoba. In total, 48 fields were sampled with fields more prone to being infested selected. Each field was
sectioned into areas that could be responsible for introduction, such as entrance ways, headlands near ditches,
depressions, drainage ways etc. The “W’ or zigzag pattern was used to sample soil in each section with the
samples composited for a section. A soil washing unit, a modified Fenwick elutriator based on the USDA soil
cyst extractor, was obtained and used in this project. The unit had an efficiency of 75 % in recovery of cysts
from samples with known amounts. In total, 282 composite soil samples were analyzed. Five lb of air-dried
soil was soaked in water and subjected to cyst extraction with the unit. Floated cysts and debris were
collected on a 60 mesh screen. The collected material was dried and then carefully scanned using
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microscopes for the presence of cysts of any nematode.
In total, 37 of the composite soil samples from 22 fields had nematode cysts. Sixty cysts, with an average of
1.6 cysts per each of the 37 composite samples were obtained. Cyst identification was performed based on
morphology and microscopy (Wouts and Baldwin 1998) and molecular analysis. Most of the cysts were
damaged but 26 were sufficiently intact for observation. Of these 26 cysts, 23 had vulval cone structures that
were circumfenestrate, consistent with belonging to the genera Cactodera, Punctodera and Betulodera. Three
cysts had bifenestrate vulval cone structures, which is consistent for the genus Heterodera. Not all the cysts
contained nematodes either as juveniles (J2 stage) or eggs. Most were empty except for 15 circumfenestrate
and one bifenestrate cysts. However, of the 26 cysts examined, 6 had eggs and J2 juveniles inside, 4 just had
eggs, and 6 just had J2 juveniles; the remaining 10 cysts were empty. These 16 cysts were then followed up
with molecular analyses by ITS sequencing, three SCN species specific PCR approaches (Ou et al. 1993,
Subbotin et al. 2001, Madani et al. 2013), and in one case, by 28s rDNA gene sequencing. ITS sequencing
was possible for 14 of the circumfenestrate cysts. These best matched the sequence database for the cyst
nematode Cactodera. Only one of the bifenestrate cysts yielded DNA for analysis. Two of three species
specific PCR approaches identified the cyst as being SCN. ITS failed to amplify for this cyst. 28s rDNA gene
amplification was successful and the sequence best matched that of Cactodera in the database. One cyst that
also failed in ITS sequencing was analyzed using 28s rDNA and it best matched Punctodera.
This survey did not find a field heavily, moderately or even lightly infested with SCN. Cactodera, and
possibly Punctodera, were found in very low levels. These are cyst nematodes but not a pest of soybean or
important pests of other crop plants in Manitoba. These cyst nematodes are likely naturally present, living on
weeds and grasses in the sampled fields. The presence of SCN is possible in one of the fields examined but
with only a single cyst and J2 juvenile recovered and the identification ambiguous between Cactodera and
SCN, further sampling and analysis is recommended for this field and also another field which had
bifenestrate vulval cone cyst structure but did not yield DNA for molecular analysis. In no way should the
results be interpreted to conclude that SCN was found in Manitoba, the results rather show the contrary. The
findings mean SCN has still not been detected in the Province. Further annual surveys are suggested using the
extraction facility setup in this current project for the next two years to encompass more soy fields and verify
the non-presence of the pest.
Objectives and Activities
2. Provide an update on all of the project objectives outlined in Schedule “B” Part I of the Agreement. If there
were any that were not met, please explain:
This project has the objectives to a) establish a SCN extraction facility and b) conduct a targeted survey of
SCN in fields most likely to be infested. Both objectives were accomplished as detailed below.
3. Provide an update on all activities outlined in the Project Work Plan in Schedule “B” Part II of the Agreement
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(insert extra lines as needed). If any of the activities have not been completed, please describe any challenges
or risks that have been encountered that have impacted the completion of the activities. Describe how you
have mitigated these risks or challenges or plan to address them going forward.
Activities
Activity One: Extraction Unit
Activity Two: Unit Setup
Activity Three: Field Sampling
Activity Four: Sample Analyses
Activity Five: Outreach
Activity Six: Administration
Final Update on Outcomes and Challenges or
Risks Encountered
After delays in funding the unit was ordered and
delivered took 2 months longer than originally
quoted
This took a month longer than expected because the
unit needed to be replumbed by us to stop water
leaks
We waited for funding to start the fall 2012 sampling
but that came through in January 2013. So we
conducted the fall sampling anyway in order to move
the project. We were able to sample 37 fields before
a very early large snow in November stopped further
sampling. The late spring 2013 prevented sampling
and sampling of 11 fields was undertaken in summer
2013. Though 2 fields less were sampled, 1100
more soil samples were taken because more
sections per field were taken. Thus the 96+% activity
completion given.
Completed in timely fashion once samples were
available. Sorting the recovered material from
extractions took longer than expected with two and
one technicians working full time and part time in the
summer months of 2013. The sorting took longer
because of the high organic carbon content of the
red river soils that made for a lot of material to be
sorted through.
Pulse Beat articles, two presentations to growers,
and three media articles were very successful. In
addition presentation to the Canadian
Phytopathology Manitoba Meeting in Nov 2013 and
distribution of presentation to MAFRD staff
The University and MPGA executed administration
of the project very well.
Completion
Date
(actual or
expected)
Degree of
Activity
Completion
(%)
30/03/2013
100%
15/06/2013
100%
15/08/2013
96+%
30/09/2013
100%
31/10/2013
100+%
15/11/2013
100%
4. Identify the project inputs used to complete the activities and during the course of the project (include:
financial contributions, other government funding, staff resources, other resources, etc.) If you did not access
all of the CAAP funding, or if your actual budget is different from the approved budget, please explain why and
outline the reason(s) for those variances. All categories that are over/under budget should be discussed:
Income
MRAC/CAAP $51,549.00
MPGA $31,000.00
Total $82,549.00
Waiting on University Budget System to be Updated and Provide Financial Report. Will forward as soon as availabl.
Expenditures
Purchase of extractors (Budgeted $6,000)
Hook up supplies (Budgeted $143.26)
Hook up and testing labour (Budgeted $7,799.85)
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Soil Sampling Labour (Budgeted $6,884.00)
Vehicle use (Budgeted $999.17)
Processing, extraction and examination of soil (Budgeted $48,860.72)
Laboratory consumables (Budgeted $4,076.00)
Administration $7,786.00 (Budgeted $7,786.00)
In-kind Contributions (not given a value)
MPGA – production of two Pulse Beat Articles and organization of one oral presentation with Dr. Tenuta
Denis Lange – identification of candidate fields (location, grower, contact and field history). Denis is with MAFRI
Mario Tenuta – project management and outreach (preparing of articles and delivery of presentations)
Soil Ecology Laboratory Facilities – stereo and dissecting microscope use, instrumentation for molecular identification
North Star Genetics – organization of one oral presentation by Dr. Tenuta
Reach and Communications/Technology Transfer
5. Identify the primary target of this project and how this has helped them adapt or remain competitive:
The primary target of this project was soybean growers in Manitoba. MPGA provided partial funding and communication
means to outreach awareness of SCN’s damage to yield, how to scout for it, control measures, and about this project’s
objectives and results to growers. MAFRI supported the project in providing grower contacts to sample fields. In all, this
was a very successful project showing the synergy of University, a commodity group, Federal and Provincial
Governments working to insure the growth of soybean acreage in the Province.
Firstly, growers are now much more aware of SCN and what it will mean to yields and change in practices when it is
found in their fields. This is a major accomplishment of the project because early detection of SCN in Manitoba means
yield losses will be lower and will slow the dispersal of the pest throughout the soy growing regions of the Province. The
extraction facility is a great asset also for future surveys of SCN. Finding no SCN in the survey is welcomed news to
growers because it simplifies their management. However, we must be vigilant and keep up awareness of the pest and
conduct future surveys.
6. Indicate the total number of people reached by this project (i.e. 10,500 producers through an electronic
newsletter, 450 attendees at a conference, circulation of 26,000 through a magazine, etc.):
Pulse Beat Spring 2013 article on the project – 500+ producers, 100+ industry
Manitoba Specialty Crops Symposium (Feb 7, 2013) – 50 producers, 25 industry
North Star Genetics Morris Producer Day (April 2, 2013) – 250+ producers
Manitoba Co-operator article print and online (Feb 12, 2013) – 5,000+ producers
Steinbach Online article (Feb 12, 2013) – 300+ producers
Grain News Article print and online (March 20, 2013) – 50,000+ producers
Pulse Beat Fall 2013 follow up article on the project findings (Fall 2013) – 500+ producers, 100+ industry
Canadian Phytopathology Manitoba Meeting (Nov 2013) – 5 industry, 25 researh
8. Indicate when you acknowledged funding provided through CAAP in accordance with Schedule “E” of the
CAAP Agreement:
MPGA and Government of Canada through MRAC and CAAP are acknowledged in interviews, presentations and
articles. Sometimes the writer neglected to list all funders. It will also be acknowledged in journal articles relating to the
project.
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Final
9. If applicable, outline how this initiative will be economically viable and self-sustaining from this point
forward and the next steps for this initiative:
Research such as tracking the presence of pest requires continued efforts. Two fields will be resampled in late fall 2013
that had bifenestrate cysts. The funding for that work will come from the Pulse Cluster 2 that is part of Growing Forward
2 of AAFC. Dr. Tenuta has a project supported by the pulse industry on surveying and determining the importance of
nematode pests of crops in Prairie Canada. As part of that project, a new method based on molecular quantification of
SCN in soil will be developed to replace the laborious hand sorting and observation required. Dr. Tenuta with Activation
Labs of Anacaster ON have applied to the AIP GF2 program for the commercialization of our methods. A survey for
SCN is recommended every 2 or 3 years to pin point when the pest builds to levels damaging to SCN. This will then
determine when growers move to management practices to combat and adapt to SCN.
10. Describe any anticipated or actual environmental and economic impact of the project (in quantifiable terms,
as outlined in the project application):
As described in the proposal, the direct economic benefits arising from the project will be difficult to measure. The
project provides a detailed information base regarding the presence of SCN in the Province through a survey of
soybean fields for the occurrence of SCN and provides a sample processing facility for continued monitoring of
the nematode in Manitoba.
Awareness of SCN to growers and industry from this project has encouraged the evaluation of SCN resistant
varieties in Manitoba. The use of these varieties, if continued to show competitive yields to non resistant varieties,
means establishment of SCN in the Province will be slowed. This should result in lower yield loses. The Thus the
project will allow Manitoba producers and seed companies to adapt to the presence of SCN and manage the pest
to insure the commodity’s growth is not restricted by this pest.
11. Provide a discussion of lessons learned, recommendations and overall perception of project success:
Lessons Learned:
The extractor had an efficiency of 75% recovery of cysts in soil
The fields sampled were free of SCN
There are other cyst nematodes present in very low levels in soils that are not SCN
The levels of these other cyst nematodes are so low that it indicates they are not pest of crops
The few cysts recovered from soils were heavily damaged. Future analyses may want to not dry and store samples but
rather conduct extractions on freshly obtained soil from the field
The technical project report following this report gives recommendations
Perception of Project Success:
Producers have been very receptive to our efforts of communicating about the project and SCN. Particularly, the
producers growing tight rotations or continuous soybeans have rightly been very interested. I am sure some of these
producers will be changing their rotation in light of this project. MPGA and MAFRI have been very pleased with the
project as it has provided a robust examination of the presence or lack of SCN in the Province, increased producer
awareness and diagnosis of SCN, and has developed capacity in Manitoba for high quality analysis of soil for SCN. The
capacity is important because suspect fields can processed here and follow up surveys can be done in Manitoba as
well.
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Media Coverage (add extra lines, if required)*
Date
Source
Title
Reach
March
2013
Feb 12
2013
MPGA Pulse Beat
SCN Coming to a Field
Near You?
Put These Crop Pests on
Your Radar
600+
Feb 12
2013
March 20
2013
Oct 2013
Steinbach Online
300+
Grain News
Soybean Cyst Nematode
Survey Underway
Soybean Cyst Nematodes
No (article was about many
pests including our work with
SCN)
MRAC
50,000+
MRAC and CAAP
MPGA Pulse Beat
SCN Survey Follow up
600+
MRAC and CAAP
Manitoba Cooperator
5,000+
CAAP Recognition
(Yes/No)
MRAC and CAAP
*If possible, please provide a copy of the media coverage for our files.
Agriculture and Agri-Food Canada (AAFC) is committed to working with industry partners. Opinions expressed in this
document are those of the Applicant and not necessarily those of AAFC
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