Oregon Wheat Commission Root-lesion Nematode Tolerance in Winter Wheat

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Oregon Wheat Commission Root-lesion Nematode Tolerance in Winter Wheat Powered By Docstoc
   (except for length, this format follows the RFP style for the Washington Wheat Commission)

Project: OSU Agricultural Research Foundation #7101 “OWC-Wheat Disease/R Smiley”

Title: Root-lesion Nematode Tolerance in Winter Wheat

Researcher: Richard Smiley, OSU-Pendleton; richard.smiley@oregonstate.edu; 541-278-4397

Cooperators: Grower Bill Jepsen and wheat breeders/pathologists Drs. Kim Kidwell, Steve Jones,
   & Steve Ullrich (WSU); Kim Garland Campbell (USDA-ARS); Jim Peterson & Pat Hayes
   (OSU); Juliet Windes & Jianli Chen (UI), Alan Dyer (MSU), and Julie Nicol (CIMMYT).

Year Initiated: July 1, 2007 (Current Year: July 1, 2008) Terminating Year: June 30, 2010

A. Goal: The goal is to screen selected PNW winter wheat varieties and advanced breeding lines to
quantify levels of tolerance to two species of plant-parasitic root-lesion nematodes. This is the
second of a three year study. This study is expected to result in an assignment of economic risk
associated with growing each winter wheat variety in soil infested with root-lesion nematodes. The
STEEP program is funding parallel studies with spring wheat and barley, and the USDA-ARS is
funding complementary experiments to identify levels of genetic resistance in spring and winter
wheat, as well as effects of crop rotations on populations of root-lesion nematodes. The OSU
Agricultural Research Foundation is funding related research to develop DNA-based tests for use in
commercial nematode diagnostic labs, and molecular markers for use in marker-assisted selection
for detection of resistance genes in seedlings.

B. Problem: Root-lesion nematodes (Pratylenchus spp.) burrow through the root epidermis and
cortex causing root pruning and damage that encourages further attack by fungal pathogens such as
Pythium and Rhizoctonia. Roots damaged by nematodes are unable to extract all available soil water
and nutrients, leading to premature onset of plant stress. Root-lesion nematode damage is very
difficult to identify and demonstrate. The first reports of root-lesion nematodes in wheat roots in the
PNW were made in eastern Washington, P. thornei in 1986 and P. neglectus in 1992. Those reports
were simple observations and greenhouse assays based on my referral of suspected samples to the
WSU nematology lab at Prosser, after I had determined that Rhizoctonia root rot could not be solely
responsible for the damage I was evaluating on several fields in Washington.
    The first indication that lesion nematodes were actually economically important in the PNW
came from a sampling of „Madsen‟ winter wheat in 1999 (Smiley et al., 2004). Madsen had been
planted uniformly at the conclusion of a 5-year crop rotation and tillage management study south
of Pendleton. There was a highly negative correlation (P < 0.0001; e.g., a 99.9% level of
confidence for this association) between lesion nematode numbers and grain yield. The
correlation coefficient (R2 = 0.64) for the regression suggested that 64% of the variation in wheat
yield at that site was associated with the number of lesion nematodes left by the previous
rotations. Unmeasured differences in soil nutrients and stored water in the treatments presumably
accounted for the remaining variability in yield.

    A similar experiment at Moro provided stronger evidence during 2007. Experimental
variables were winter wheat rotated with cultivated or chemical summer fallow, no-till annual
winter wheat, spring wheat, and spring barley, no-till winter wheat/winter pea rotation, and a 3-
year rotation of no-till winter wheat, no-till spring barley, and chemical fallow. Each phase of
each rotation occurs each year. Several flexible-decision treatments, based on factors such as
weather and markets, are also planted to crops such as spring mustard, barley, or wheat. A highly
significant (P < 0.0001) negative correlation occurred between grain yield and number of lesion
nematodes (Figure 1). Nearly all (94%) of the variability in winter wheat yield at that location,
from 2005 to 2007, was attributable to the numbers of root-lesion nematodes in soils of each
experimental treatment. The remaining 6% of variability was presumably due to variations in
weed density and availability of nutrients and water. Soil water was left unused in plots with high
nematode counts. Populations of lesion nematodes were high following crops of winter wheat,
spring wheat, winter pea and spring mustard, and lower following spring barley or summer
fallow (Table 1). This study confirmed that 1) the type of fallow has little or no effect on
nematode numbers, 2) wheat attains higher yield potential following barley compared to a
previous crop of wheat, and 3) barley reduces nematode numbers in soil and therefore the level
of risk for subsequent crops that are intolerant of root-lesion nematodes.
    Two recent surveys were conducted to determine if lesion nematode populations were high
enough to warrant detailed research in the PNW (Smiley et al., 2004; Strausbaugh et al., 2004).
These nematodes were found in 95% of the 281 fields sampled in 23 Idaho, Oregon and
Washington counties. P. neglectus was the most prevalent species in each state. We now estimate
that about 60% of all PNW wheat fields harbor potentially damaging populations of root-lesion
nematodes. These fields are distributed across the region (Figure 2) but we have also detected a
few areas where the populations are notably low, such as in several areas of Adams County, WA.
    Crop management options other than genetic tolerance and resistance are not likely to be
economically feasible for most dryland wheat producers in low-rainfall regions. Chemicals are
neither registered nor economically feasible in dryland agriculture. Tillage intensity does not greatly
affect populations of root-lesion nematodes. The frequency of favorable host plants can be
minimized by not allowing volunteer cereals and winter-annual weeds to grow during the winter
between planted crops; e.g., the green bridge must be totally eliminated. Crop rotations may be
helpful, detrimental, or neutral. Populations of these nematodes can be reduced by crops such as
barley, field pea and safflower, and are increased by chickpea. Most canola, mustard and lentil
varieties appear to promote reproduction of P. neglectus but possibly not P. thornei.
    Australian spring wheat varieties with known levels of tolerance to root-lesion nematodes were
studied in annual direct-drill systems in Oregon from 2001 to 2004 (Smiley et al., 2005a, 2005b).
Tolerance is a field-based measure of the ability of a crop to maintain grain yield under high
nematode pressure. Grain yield for moderately tolerant varieties was up to double that for intolerant
varieties in highly-infested fields at Moro and Pendleton, OR. Increasing densities of root-lesion
nematodes were strongly associated with reduced yield of intolerant varieties. However, none of the
affected crops exhibited a definitive symptom of injury that could be considered diagnostic for this
problem. Plants of sensitive (intolerant) varieties were simply thinner and less vigorous than plants in
adjacent nematicide-treated plots that yielded twice the amount of grain.
    The first evaluations of wheat and barley varieties and advanced breeding lines adapted to the
PNW were performed during 2006 and 2007. These experiments evaluated the influences of P.
neglectus and P. thornei on yields for 79 wheat and 11 barley entries; 45 of which were planted

during spring and 45 during the fall. Entries included 24 advanced breeding lines and 57
commercial varieties representing recent production on 95% of the winter cereals acreage and
85% of spring cereals acreage in Oregon and Washington.
         When data were averaged over the two years of testing, P. neglectus-tolerant spring wheat
varieties such as Tara 2002 and Jerome had little or no yield reduction, and intolerant entries
such as IDO377S and OR4201019 yielded about 40% less grain than the most-tolerant varieties
(Figure 3). The tolerance index of 96% for Tara 2002 indicated that the nematicide improved
grain yield by only 4%. The tolerance index of 60% for IDO377S indicated that this nematode
suppressed yield by 40%. In the presence of P. thornei, the most tolerant entries included
Wawawai, Jefferson, Otis, Tara 2002, Zak, Buck Pronto, Jerome and Louise (data not shown).
The least tolerant varieties were Eden, McNeal, and Calorwa.
    Yields of spring grains in untreated soil were negatively correlated with the tolerance index
(P < 0.0001). The high correlation coefficients in both P. neglectus- and P. thornei-infested soil
(R2 = 0.79 and 0.70) suggested that 79% and 70% of the yield variability at these locations was
associated with the number of nematodes. This likelihood was supported and strengthened by the
fact that each of the entries in these tests yielded relatively the same amount of grain if the soil
was treated with the nematicide at the time of planting (Figure 3).
    Unfortunately, as previously was demonstrated overseas, most spring wheat varieties and
lines reacted differently to P. neglectus and P. thornei (Figure 4). The correlation coefficient
between tolerance indices for these nematodes was very low (R2 = 0.16), indicating little
correlation between these tolerance reactions. Tests must continue to be conducted at two
locations. Fortunately, varieties such as Tara 2002, Jerome, Otis and Louise exhibited a high
tolerance to both nematode species, making them good choices to plant in any infested field, but
particularly in fields where mixtures of the two species occur.
    Spring barley yields were higher than spring wheat yields in fields infested by both nematode
species (see data for P. neglectus in Figure 3), confirming previous observations that spring barley is
generally more tolerant than spring wheat. Barley also suppresses the multiplication of P. neglectus,
resulting in a lower nematode population following a barley crop.
    Yields of winter wheat entries were not closely correlated to the tolerance index. The large
nematicide and varietal effects we have observed in individual plots have not yet withstood the rigors
of replicated and multi-year testing. For P. neglectus (Figure 5), only 14% of the yield variation
among fall-planted entries could be directly attributed to damage by this nematode. We believe that
the narrower spread of tolerance index numbers for fall- than spring-planted entries is an artifact of
our testing protocol. The nematicide used for these studies has an effective life no longer than two
months, which represents about 50% and 20% of the growing seasons for spring- and fall-planted
cereals, respectively. In a small auxiliary experiment with five winter wheat entries during 2006-
2007, the precision of our test was doubled when the nematicide was applied twice rather than only
at the time of planting. We have therefore refined the testing procedure for the 2007 fall-planted
experiments in an attempt to achieve a level of precision nearer to that for our spring-planted
nurseries. Rigorous additional screening will be required before we can confidently describe
tolerance levels for the winter wheat entries.
    All spring- and fall-planted entries tested for field tolerance were also screened for resistance.
Resistance is a measure of the ability of the nematode to multiply after it enters the root, and is
studied in the greenhouse. Resistant varieties allow little or no multiplication of the nematode,
leaving fewer nematodes and a reduced risk for future crops. Susceptible varieties allow

expansive multiplication. Resistance and tolerance mechanisms are genetically unrelated. A
given variety can be tolerant and resistant, tolerant and susceptible, intolerant and resistant, or
intolerant and susceptible. These traits are controlled by multiple genes, making it possible for a
large group of varieties to exhibit a continuous range in levels of tolerance as well as resistance.
    All PNW wheat varieties and breeding lines were found to be susceptible to both species of
Pratylenchus; see data for P. neglectus in Figures 6 and 7. All spring and winter wheat varieties
and lines allowed expansive multiplication of these nematodes and, therefore, are likely to
increase the risk for damage to subsequent plantings of intolerant crops and varieties.
Fortunately, several exotic resistant wheat entries used as controls, based on previous research in
Australia, were highly resistant in our tests. There was no correlation between the resistance and
tolerance ratings for these 45 spring (R2 = 0.01) and 45 winter (R2 = 0.04) wheat entries.
    We are introducing genetic resistance into locally-adapted germplasm. Six exotic wheat lines
carrying highly effective resistance genes have been crossed with nine PNW varieties or lines;
Louise, Otis, Alpowa, Stephens, Tubbs 06, Brundage 96, Goetze, ORSS1757, and ORH010085.
Some crosses were made to pyramid resistance genes, in an attempt to provide protection against
both species of lesion nematode, the cereal cyst nematode, and Fusarium crown rot (foot rot).
Resistant adapted lines will be transferred to wheat breeding programs in each state.
    In collaboration with USDA-ARS scientists at Pullman, we are also developing molecular
markers to increase the efficiency of tracking these resistance genes in breeding programs, and
DNA-based tests to increase the speed and accuracy for quantifying and identifying the presence
of these nematodes in soil. These new testing protocols will be transferred to breeders and
nematode diagnostic laboratories within two years.
    Selecting and breeding wheat for tolerance and resistance is very likely to improve productivity
and economic efficiency where-ever susceptible host plants dominate the cropping system, including
each crop in the rotation and also the grass and broadleaf weeds. Research proposed here aims to
quantify tolerance levels for PNW winter wheat varieties and advanced breeding lines. Parallel
studies with spring wheat and barley are being funded by the STEEP program, and genetic resistance
studies are being performed as part of our contract with the USDA-ARS. Together, these studies are
intended to provide a comprehensive package of information on genetic tolerance and resistance,
crop rotations, and tools to enhance the capabilities of breeders and commercial diagnostic labs.

References to research with root-lesion nematodes on wheat in the PNW:
1. Smiley, R.W., K. Merrifield, L.-M. Patterson, R.G. Whittaker, J.A. Gourlie, and S.A. Easley. 2004.
   Nematodes in dryland field crops in the semiarid Pacific Northwest United States. Journal of Nematology
2. Strausbaugh, C.A., C.A. Bradley, A.C. Koehn, and R.L. Forster. 2004. Survey of root diseases of wheat
   and barley in southeastern Idaho. Canadian Journal of Plant Pathology 26:167-176.
3. Smiley, R.W., R.G. Whittaker, J.A. Gourlie, and S.A. Easley. 2005a. Pratylenchus thornei associated with
   reduced wheat yield in Oregon. Journal of Nematology 37:45-54.
4. Smiley, R.W., R.G. Whittaker, J.A. Gourlie, and S.A. Easley. 2005b. Suppression of wheat growth and
   yield by Pratylenchus neglectus in the Pacific Northwest. Plant Disease 89:958-968.

C. Background/Scope:

    1) Location of the problem or number of acres affected: Throughout the region; not restricted by
precipitation zone, temperature, or soil type (see Figure 2). Most prevalent in annual crop systems
including “functionally annual crops” where volunteer cereals and winter-annual weeds are present

during the “fallow” winter in winter wheat-summer fallow rotations.

   2) Classes and/or varieties affected: All classes of wheat.

    3) Other work conducted on this same issue in the PNW and the US: There is no comparable
research in the USA although results of our studies in Oregon have now stimulated interest in this
topic in at least five other western states; ID, CO, KS, MT and WA. The PI receives funds from an
OSU subcontract to the USDA-ARS (SCA #58-5348-9-100, “Control of Root Diseases of Wheat and
Barley”) to perform research on strategies to manage wheat and barley root diseases caused by
Fusarium crown rot, root-lesion nematodes, and cereal cyst nematodes. The funding level for the
ARS grant was reduced 25% during the current fiscal year and future funding is uncertain. Current
funds are sufficient to finance one-third of the tolerance and resistance research proposed here for
FY2008. Leveraging is also achieved through affiliated molecular studies funded by the OSU
Agricultural Research Foundation.

    4) Preliminary studies showing research is feasible: Successive stages of this topic have been
actively researched at Pendleton since 1999 (see “Goals” and “Problem”). Our earlier research
protocols have each been endorsed through the peer review process during publication of technical
journal papers.

    5) Reason for addressing the problem now: Our research and the surveys performed in all three
states have clearly demonstrated a new understanding of the breadth and extent of this nematode
problem in the PNW. Recent results from the crop rotation and tillage management study at Moro
(Figure 1 and Table 1) are remarkable in that the root-lesion nematodes at that location (a mixture of
both species) appear responsible for 94% of the yield variation in winter wheat, regardless of the
length or type of crop rotation or tillage management.

D. Economics: Tests in Oregon have shown that yields of intolerant varieties can sometimes be
doubled by application of a non-registered nematicide, and that yields of moderately tolerant
varieties can be nearly double the yields of intolerant varieties, even though varieties of both types
have comparable yields on fields with low populations of root-lesion nematodes. Based on the
amount of acreage planted to each spring wheat variety in Oregon and Washington, and assuming
that only 60% of the acreage is infested with a high number of root-lesion nematodes, we have
calculated that wheat yields could be improved 15% to 20% region-wide if all acreage was planted to
a variety as tolerant as Tara 2002. Likewise, even though our measurements with winter wheat are
still much poorer than for spring wheat, the comparable calculation suggests that regional winter
wheat yields could be improved 5% if only the most tolerant varieties were planted. However, yields
on some individual fields could be improved as much as 40%.

A. Objective: Characterize levels of field tolerance to Pratylenchus neglectus and P. thornei in
   Pacific Northwest winter wheat varieties and advanced breeding lines.

B. Procedures: Genetic tolerance measures a plant‟s ability to yield acceptably well even when
invaded by moderately high populations of root-lesion nematodes. The tolerance index is calculated

from the ratio of grain yields in untreated and in nematicide-treated soil. As discussed earlier, it
became apparent that our nematicide application procedure needed to be improved for fall-planted
nurseries. For P. thornei, we will continue to test the selection of 39 winter wheat entries examined
during the past two years (Fig. 5), plus four additional varieties added by special request from the
industry in southeast Idaho. For P. neglectus, we have increased the number of entries to 121,
including additional commercial varieties and advanced breeding lines supplied by Drs. Jim Peterson
and Pat Hayes (OSU), Kim Campbell (USDA-ARS), Steve Jones (WSU), Juliet Windes and Jianli
Chen (UI), Alan Dyer (MSU), and Julie Nicol (CIMMYT- Turkey).
    Our P. neglectus-infested site is at the Bill Jepsen Farm between Heppner and Condon. The P.
thornei-infested site is at the Agricultural Research Center near Pendleton. Fields at both locations
are managed without primary tillage. The Jepsen fields have a history of annual direct-drill spring
crops including barley, wheat and mustard. The Pendleton fields are in a three year-rotation of spring
barley, winter wheat, and chem-fallow. Tests may follow any of the crops at the Jepsen farm and the
chem-fallow at Pendleton. Grid sampling is performed in prospective fields during May or June 2007
to identify specific test sites with high and uniform populations of these nematodes.
    During September 2008 the test sites will be measured and staked, sprayed with glyphosate if
necessary to kill volunteer cereals and grass weeds, and fertilized at rates based on soil testing and
anticipated yield potential. Fertilizer will be applied using an 8-foot wide Fabro drill with double-
disk openers at 10-inch spacing. In September or October 2008 the wheat entries will be planted
without tillage using a John Deere HZ drill with openers at 14-inch spacing, a cone seeder for
dispensing seed, and a Gandy box for dispensing Temik below the seed. Each wheat entry is planted
using a strip-plot design with 6 x 30-foot plots replicated 3 times. Temik-treated and control plots for
each entry are planted in adjacent drill strips, allowing side-by-side comparison of treatments for
each entry. The experiments will have at least 240 plots; a minimum of 40 wheat entries x 3
replicates x 2 chemicals (none or Temik).
    Seedling emergence and plant growth will be evaluated as necessary on each experimental area.
During spring, a broadleaf weed spray will be applied and the nematicide will be reapplied as an in-
furrow top-dressing in treated plots. Plots will be harvested using a small plot combine and grain
yield and test weight will be calculated. The root-lesion nematode tolerance index for each entry will
be calculated as the ratio of grain yield in control and Temik-treated plots. Data will be analyzed
using ANOVA or REML statistical procedures. Results will be compared to data collected during
2006 and 2007. All grain from these experiments must be destroyed due to our use of the nematicide.

C. Cooperation and Coordination: Plant materials for this research have been and will continue to be
supplied by wheat and barley breeders throughout the PNW and by the CIMMYT/ICARDA Root
Disease Testing Nursery program (see the list of cooperators for this grant). Base-level funding is
provided by a USDA-ARS research program that includes sub-contracts to the University of Idaho
(Dr. Juliet Windes, Idaho Falls) and Oregon State University (PI for this proposal). The USDA
funding also provides support for scientists in the parent USDA-ARS program, including Drs. David
Weller, Tim Paulitz, Linda Thomashow, and Patricia Okubara. The principal investigator for this
proposal is collaborating with Dr. Okubara in screening the novel Scarlet Rz! selection, and in
developing DNA-based diagnostic tests for commercial soil testing labs. He also collaborates with
Dr. Campbell in screening germplasm for tolerance to Fusarium crown rot, and in developing
molecular markers to detect resistance genes for the cereal cyst nematode. The PI for this study also
serves with Drs. Paulitz and Campbell, and previously also with Dr. Kidwell, on the dissertation

committees for graduate students at WSU, including a student currently focusing on Fusarium crown
rot. Finally, the PI also provided specific guidance for the earlier survey of fungal pathogens and
nematodes in southeast Idaho, as well as for current studies of Fusarium crown rot in that region.

D. Review: The principal investigator participates in 15 to 25 public presentations annually. Venues
are mostly in Oregon and Washington although periodic invitations also come from Idaho. Speeches,
written proceedings, and/or posters are presented at experiment station field tours, extension service
crop tours and training schools, meetings of agricultural consultants, training schools or farmer
meetings coordinated by agricultural chemical companies, professional conferences, industry
conferences such as PNW Direct-Drill, guest lectures and seminars at universities, and research
reviews by funding agencies such as STEEP and OWC. If asked to do so, the PI also anticipates
and/or expects to appear at venues sponsored by the WWC and/or IWC, or by the affiliated grain-
industry organizations in those states.

E. Locations: Plant tolerance and resistance are plant characteristics that are equally expressed across
environments. To maintain continuity of cost-effective research, these tolerance experiments are at
two locations where appropriately high populations of the two nematode species are already well
defined. P. neglectus is studied at a commercial farm between Heppner and Condon, OR (12-inch
rainfall, 2,550-ft elevation). P. thornei is studied at the Columbia Basin Agricultural Research Center
near Pendleton (16-inch rainfall, 1,470-ft elevation).


A. Expected Results: Productivity of winter wheat on fields infested with high populations of root-
lesion nematodes will be improved (see Figure 1). Results of this research are already being
communicated to producers, breeders, extension agents, commercial nematode diagnostic labs, and
grower-advisors from agribusiness service and supply companies. Methods of communication
include 1) participating in industry and professional meetings (direct-drill, state grains organizations,
county crops tours and winter meetings, experiment station field days, Far West Agribusiness, PNW
Crop Consultants, and professional meetings of pathologists, nematologists, and crop scientists), 2)
writing experiment station publications and wheat-industry magazine articles, 3) writing a PNW
extension bulletin, and 4) publishing technical journal manuscripts. The PI also intends to submit
results of this work for listing in seed-buyers guides. Demonstration plots will be shown and/or
discussed during field tours. Although not part of this project, on-going testing would then become
focused on elite breeding lines during the three years prior to their release. These studies are
complemented by research to 1) introduce resistance genes into PNW-adapted varieties, 2) develop
molecular markers to genes for resistance to P. thornei and P. neglectus in seedlings, and 3) develop
molecular diagnostic tests to differentiate and identify P. neglectus and P. thornei in field soils.

B. Payback Timeline: Assignment of tolerance indices for winter wheat is anticipated within three
months after this experiment is terminated in June 2010. Identification of genotypes with superior
resistance has already occurred and crosses are being made with PNW-adapted germplasm. The
progeny of those crosses will be backcrossed again to the source of resistance and the progeny will
be screened to identify genotypes carrying the resistance genes. At least one wheat breeders has
already requested and received progeny of the first crosses.

C. Progress: Two seasons of field tolerance testing have been completed. Our inability thus far to
attain the level of precision achieved with spring grains has caused us to make further refinements in
the nematicide application process. We anticipate greater clarity of separation among tolerance levels
for the winter wheat entries during the upcoming season. The resistance testing in the greenhouse has
shown that all of the winter and spring wheat varieties and breeding lines in the PNW are
susceptible. There is no need to continue pursuing that line of investigation. Instead, we have moved
directly into the process of introducing highly effective resistance genes from Iranian landrace lines
into PNW-adapted wheat varieties. Seed from the BC1F1 generation has been harvested.

D. Impact: Smiley et al. published results showing that yields of tolerant spring and winter wheat
varieties had yields as much as double those for intolerant varieties. Thus far, results of the PI‟s
findings have withstood the “test of time” through further scrutiny by scientists in other states and
overseas. Also, growers have indicated to the PI that this research appears to explain why their wheat
yield is better following barley than canola, mustard or chickpea, when water is not responsible for
those observations. Accurate economic estimates have not been attempted but the value of this
research will surely increase as annual cropping and especially oilseed crops become more common.

E. Communication: Please refer to the “Process: Review” section for this explanation.


A. Amount allocated by the Tri-State Commissions in FY 2007-08: $24,969

B. Request for FY 2008 (July 1, 2008-June 30, 2009):
Refer to Addendum 2 to view the anticipated full budget for this genetic tolerance and resistance
project for the 3-year period FY2007-FY2009. Note that the budget proposed to the Tri-State
Commissions is for the winter wheat component of this study and assumes that equal funding for
parallel research has already been allocated by the STEEP program for spring wheat and barley. The
budget also shows funds anticipated from the OSU sub-contract to the USDA-ARS Biological
Control and Root Disease Research Laboratory, at Pullman. The USDA-ARS sub-contract is also
expected to also provide adequate funds to include research on Fusarium crown rot and cereal cyst
nematode. The OSU-Agricultural Research Foundation is providing funds to develop molecular
markers for root-lesion nematodes, and to develop molecular procedures to identify nematode
species in soil.

            Salaries                           $10,482
            Time-slip                            2,277
            Employee benefits                    8,673
            Goods and services                   1,200
            Travel: research plots                 961
            Travel: reporting/reviews            1,376
            Equipment                                0
            Total                              $24,969

Addendum 1. Timetable for Tolerance Testing for PNW Winter Wheat

Objective #1 - Preparation (not included in budget):
   May-June 2007          Pre-plant grid sampling of soils on potential trial sites for 2007. Measure
                              soil moisture and store samples for processing
   Sep 2006-June 2007 Preliminary experiments to refine Temik application procedures with
                              winter wheat
Objective #1 - Year 1 of grant:
   July/Aug 2007          Extract and quantify nematodes from pre-plant sampling, re-sample if
                              necessary, identify 2007 fall-planted and 2008 spring-planted sites
   July-Aug 2007          Harvest the 2006-2007 preliminary experiment
   Aug-Oct 2007           Process 2007 harvest samples, analyze data, & destroy all harvested grain
   September 2007         Fertilize and spray Roundup at each trial site
   October 2007           Plant nurseries at Pendleton (P. thornei) and Heppner (P. neglectus)
   Oct 2007-Feb 2008 Evaluate plant growth multiple times, reapply nematicide to each trial
   Mar-July 2008          Spray broadleaf weeds and evaluate plant growth multiple times
   June 2008              Pre-plant grid sampling of soils on potential trial sites for 2008-2009
   Throughout             Communicate updated observations and results to appropriate audiences

Objective #1 - Year 2 of grant: (the current phase of this project)
  July-Aug 2008          Harvest the 2007-2008 nurseries
  Aug-Oct 2008           Process 2008 harvest samples, analyze data, & destroy all harvested grain
  September 2008         Fertilize and spray Roundup on each trial site
  October 2008           Plant nurseries at Pendleton (P. thornei) and Heppner (P. neglectus)
  Oct 2008-Feb 2009 Evaluate plant growth multiple times, reapply nematicide to each trial
  Mar-July 2009          Spray broadleaf weeds and evaluate plant growth multiple times
  May-June 2009          Pre-plant grid sampling of soils on potential trial sites for 2009. Measure
                             soil moisture and store samples for processing
  Throughout             Communicate updated observations and results to appropriate audiences

Objective #1 – Year 3 of grant:
   July-Aug 2009          Harvest each of the four 2008-2009 nurseries
   Aug-Oct 2009           Process 2009 harvest samples, analyze data, & destroy all harvested grain
   September 2009         Fertilize and spray Roundup on trial sites
   October 2009           Plant nurseries at Pendleton (P. thornei) and Heppner (P. neglectus)
   Oct 2009-Feb 2010 Evaluate plant growth multiple times, reapply nematicide to each trial
   Mar-July 2010          Spray broadleaf weeds and evaluate plant growth multiple times
   Throughout             Communicate updated observations and results to appropriate audiences

Objective #1 - Year 3 of grant (not included in budget):
   July-Aug 2010          Harvest the 2000-2010 nurseries
   Aug-Oct 2010           Process 2010 harvest samples, analyze data, & destroy all harvested grain

Addendum 2: Budget to screen wheat for tolerance and resistance to root-lesion nematodes; July 1, 2007 to June 30, 2010.

  Budget for Screening: Updated 11/26/2006                         Tolerance: Winter wheat    Tolerance: Spring cereals     Resistance: All wheat
                                                                      FY        FY       FY      FY        FY        FY      FY        FY         FY
                                                                    2007      2008     2009    2007      2008      2009    2007      2008      2009

                                                                     request to Tri-State              STEEP                    USDA-ARS
                                                                     wheat commissions                 request                 funds in-hand
  Salaries and wages
  Professional (academic ranks) (salary X days activity)
      Sub-total for professional personnel                         10,828   10,482   10,613   11,501   10,482    10,613    6,578   13,965       8,382
  Support Personnel (classified & temporary)
      Sub-total for support personnel                               2,077    2,277    1,988    2,077    3,277     3,077    2,442    3,442       1,942
  Total wages and salaries                                         12,905   12,759   12,602   13,578   13,759    13,690    9,020   17,407      10,324
  Employee benefits                                                 8,758    8,673    8,626    8,961    8,923     8,913    7,834   10,318       8,255
  Total salaries, wages, & benefits                                21,663   21,432   21,227   22,539   22,682    22,603   16,855   27,725      18,579
  Goods and Services
      Tolerance: chemicals, fertilizers, bags, flags, stakes        1,000    1,200    1,200     900     1,000     1,000
      Resistance: greenhouse electricity & propane, bags                                                                   7,250    7,250        100
    Industry reviews (OWC, WWC, IWC?, OWGL)
       Lodging, food and registration (days x $130/day)              780       780      780     260       260      260
       Mileage: miles x 0.445/mile                                   596       596      596      89        89       89
    Travel related to servicing experimental plots
       3/4T or 1T, often with light trailer: miles x 0.445/mile       561      641      641      481      481       481
       2.5T truck with heavy trailer: miles x 0.89/mile               320      320      320      160      160       160
  Equipment                                                             0        0        0        0        0         0
  Total Expense to all potential sponsors                          24,920   24,970   24,765   24,429   24,672    24,593   24,105   34,975      18,679

                                                                       FY       FY       FY
  Tolerance and Resistance Screening                                 2007     2008     2009
  Total Expense                                                    73,454   84,616   68,037
  Tolerance expense for winter wheat (OWC+WWC+IWC)                   34%      30%      36%
  Tolerance expense for spring wheat and barley (STEEP)              33%      29%      36%
  Resistances for all cereals (OSU contract to USDA-ARS              33%      41%      27%

Figure 1. Relationship between root-lesion                              Figure 2. Distribution of survey sites where root-
nematode populations (RLN; expressed as the log                         lesion nematodes exceeded the estimated
transformed number/kg of soil) and yields for                           threshold for economic damage; reports of
winter wheat in six different crop rotations and/or                     surveys in Oregon and Washington during 1999-
tillage management systems; data are the                                2000 (Smiley et al., Journal of Nematology, 2004)
averages over three years (crop years 2005-2007)                        and in Idaho during 2001-2002 (Strausbaugh et
for each treatment.                                                     al., Canadian Journal of Plant Pathology, 2004).

                                  y = -24.015x + 215.27
                                        R2 = 0.9427
                  50                     P < 0.0001
  Yield (bu/ac)

                       6        7              8               9
                               RLN: (ln+1)/kg soil

Table 1. Population of root-lesion nematodes
(Pratylenchus spp./lb of soil) in soil during early
spring (March 7 - April 4) following specific crops                     Figure 3. Spring wheat and barley tolerance index
or management practices over a 3-year period in                         for Pratylenchus neglectus (bars) and grain yields
the long-term experiment at Moro, Oregon. Winter                        (lines; treated = ▲, untreated = ■); average of
wheat varieties were Tubbs in 04-05, Stephens in                        tests in 2006 and 2007 near Heppner, OR.
05-06, and ORCF 101 in 06-07. Spring wheat                                                 100                                                                                                                                                                                                                                                                                                                                                                                                                         70

varieties were Zak in 04-05 and Louise in 05-06                                            90

and 06-07. The number of plots “n” sampled

varied for each designated sequence over the 3-                                            70                                                                                                                                                                                                                                                                                                                                                                                                                          50

year period for crop years 2005 to 2007.
                                                                         Tolerance index

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            Yield (bu/ac)

              Previous                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 30
                                           Prats/lb       n                                40
                                                                                           30                                                                                                                                                                                                                                                                                                                                                                                                                          20

                                                                                           20                                                                                                                                                 wheat
              Winter wheat                      907 a     45                               10

              Mustard cv Tilney                 834 ab     3                                0                                                                                                                                                                                                                                                                                                                                                                                                                          0
              Spring wheat                      740 ab    15










                                                                                                 Tara 2002



                                                                                                                                                                                                                             Yecora Rojo

                                                                                                                                                      Buck Pronto







              Winter pea cv Spector             700 ab     9
              Spring barley cv Camas             451 b    21                                                                                                                                                                                                            Variety or line
              Conventional fallow                373 b     9
              Chem fallow                        353 b    24
              p>F                              0.0292*
              CV (%)                               16.0

                   Figure 4. Lack of correlation between tolerance                                                                                                                                  Figure 6. Susceptibility ratings for spring wheat to
                   indices (TI) for 40 spring wheat entries growing in                                                                                                                              Pratylenchus neglectus. The reproductive factor
                   soils infested with either P. neglectus or P. thornei                                                                                                                            (Rf) compares the final and initial nematode
                   during 2006 and 2007, showing the need to                                                                                                                                        populations following 16 weeks of plant growth in
                   perform independent screenings for each                                                                                                                                          the greenhouse. A rating of 60 indicates an 60-fold
                   nematode species.                                                                                                                                                                increase (from 432 initially to 25,920 at the end of
                                                                                                                                                                                                    the test) in the Pratylenchus population for that
                                               100                                                                                                                                                  variety. A rating of 1 indicates a lack of
                                                                                                                                                                                                    reproduction, with the final and initial populations
                                                                R2 = 0.1643                                                                                                                         being equal.
                                                                P = 0.0174
                           TI for P. thornei



                                                                                                                                                                                                                                   Reproduction of P. neglectus
                                                                                                                                                                                                          40                        (Rf = final/initial population)


                                                70                                                                                                                                                        20
                                                     50                     60                 70            80                           90                     100
                                                                                              TI for P. neglectus









                                                                                                                                                                                                                Buck Pronto




                                                                                                                                                                                                                 Tara 2002

                                                                                                                                                                                                                  Persia 20





                                                                                                                                                                                                               Yecora Rojo





                   Figure 5. Winter wheat and barley tolerance index
                   for Pratylenchus neglectus (bars) and grain yields
                   (lines; treated = ▲, untreated = ■); average of                                                                                                                                  Figure 7. Susceptibility ratings for winter wheat to
                   tests in 2006 and 2007 near Heppner, OR.                                                                                                                                         Pratylenchus neglectus. The reproductive factor
                                                                                                                                                                                                    (Rf) compares the final and initial nematode
                  100                                                                                                                                                     60
                                                                                                                                                                                                    populations following 16 weeks of plant growth in
                                                                                                                                                                          50                        the greenhouse. A rating of 80 indicates an 80-fold
                                                                                                                                                                                                    increase (from 432 initially to 34,560 at the end of
Tolerance index

                                                                                                                                                                               Yield (bu/ac)

                  60                                                                                                                                                                                the test) in the Pratylenchus population for that
                  50                                                                                                                                                      30                        variety. A rating of 1 indicates a lack of
                                                                                                                                                                                                    reproduction, with the final and initial populations

                                                                                        wheat                                                                                                       being equal.
                   0                                                                                                                                                      0
                                                                                                                                                                                                                                 Reproduction of P. neglectus









                                                                                                                                                 Stab 113
                                                                                                                                                              88 Ab 536
                                                              Idaho 587


                                                              Tubbs 06

                                                                          Brundage 96



                                         Golden Spike





                                                                                                                  Westbred 528

                                                                                                                                                                                                                                  (Rf = final/initial population)

                                                                                           Variety or line                                                                                                40




                                                                                                                                                                                                                Golden Spike







                                                                                                                                                                                                                   Idaho 587


                                                                                                                                                                                                                    Tubbs 06

                                                                                                                                                                                                                    Persia 20





                                                                                                                                                                                                                Brundage 96
                                                                                                                                                                                                               Westbred 528


                                                                                                                                                                                                                  88 Ab 536

                             Current and Pending Support

Name      Supporting      Total $    Effective &        % of Time   Title of Project
          Agency          amount     Expiration Dates   Committed
Richard   USDA-ARS        $121,856   7/7/2007 -         40%         Control of root diseases
Smiley    (subcontract)              7/6/2008                       of wheat and barley
Richard   Hatch (aka      $ 74,612   10/1/2007 -        15%         Root-lesion nematode
Smiley    STEEP)                     9/30/2008                      tolerance in spring
                                                                    wheat and barley
Richard   Tri-State   $ 24,920       Pending for        15%         Root-lesion nematode
Smiley    Commissions                FY2008 (this                   tolerance in winter
                                     proposal)                      wheat
Richard   Agric. Res.     $12,500    7/1/2007 -         3%          Developing a molecular
Smiley    Foundation                 6/30/2009                      diagnostic method to
          (PI: Guiping                                              identify root-lesion
          Yan)                                                      nematodes in soil
Richard   Agric. Res.     $12,500    7/1/2007 -         3%          Molecular markers for
Smiley    Foundation                 6/30/2009                      breeding wheat with
          (PI: Guiping                                              resistance to root-lesion
          Yan)                                                      nematodes
Richard   USDA-           $18,600    Pending:           5%          Reaction of Scarlet Rz1
Smiley    CSREES-                    7/7/2008 -                     to Pratylenchus and
          SARE (PI:                  7/6/2011                       Fusarium
          Pat Okubara)
Richard   USDA +          $15,000    Pending:           1%          Developing camelina as
Smiley    US-DOE (PI:                10/1/2008 -                    an oilseed feedstock
          Russ Karow)                9/30/2010

                                   Dr. Richard Smiley (CV)

                   Professor of Plant Pathology, Oregon State University
                        Columbia Basin Agricultural Research Center,
                                     Pendleton, Oregon

  California State Polytechnic University, 1965, B.Sc. in Soil Science
  Washington State University, 1969, M.Sc. in Soils, with a Plant Pathology minor
  Washington State University, 1972, Ph.D. in Plant Pathology, with a Soils minor

Professional Positions and General Duties:
   1. Assistant Research Soil Scientist, USDA-Agricultural Research Service, Pullman,
       Washington, 1966-1969: Research on the etiology of a lucerne-decline phenomenon,
       and methods for utilizing the fungitoxicity of anhydrous ammonia fertilizer to assist in
       controlling Fusarium diseases of wheat and green processing pea.
   2. Research Assistant in Plant Pathology, Washington State University, 1969-1972:
       Research on the differential effects of ammonium- and nitrate-nitrogen on severity of
       take-all on wheat, with emphasis on rhizosphere pH and diffusion or flow of nutrients.
   3. Research Officer in Soil Microbiology, CSIRO, Adelaide, Australia, 1972-1973:
       Research on the microbial ecology of the wheat rhizosphere in relation to infectivity by
       the take-all pathogen, Gaeumannomyces graminis var. tritici. Emphasis was on root-
       colonizing species and biotypes of Pseudomonas.
   4. Assistant (and then Associate) Professor of Plant Pathology, Cornell University,
       Ithaca, New York, 1973-1985: Research and extension on the etiology and control of
       turfgrass and wheat diseases, and on microbiology of turfgrass ecosystems. Emphasis
       was on turfgrass patch diseases caused by the soilborne pathogens Leptosphaeria korrae
       and Magnaporthe poae.
   5. Visiting Senior Scientist, Plant Research Institute, Victoria Department of
       Agriculture, Melbourne, Australia, 1982-1983: Research on the etiology and control of
       a root rot complex of subterranean clover, including species of Pythium and Rhizoctonia.
   6. Professor and Superintendent, Columbia Basin Agricultural Research Center,
       Oregon State University, Pendleton, Oregon, 1985-2000: Conduct research and
       extension on the etiology and control of field crop diseases, with emphasis on integrated
       management strategies for wheat and barley diseases caused by soilborne plant-
       pathogenic fungi (Rhizoctonia, Pythium, Fusarium, Bipolaris) and plant-parasitic
       nematodes (Heterodera, and Pratylenchus). Administer two experiment stations
       (Pendleton and Moro), including oversight for research and extension programs in soil
       science, crop science, weed science, wheat breeding and plant pathology.
   7. Professor, Columbia Basin Agricultural Research Center, Oregon State University,
       Pendleton, Oregon, 2000-present: Conduct research and extension on the etiology and
       control of field crop diseases, with emphasis on integrated management strategies for
       wheat and barley diseases caused by soilborne plant-pathogenic fungi (Rhizoctonia,
       Pythium, Fusarium, Bipolaris) and plant-parasitic nematodes (Heterodera, and

   Professional Societies:
   American Phytopathological Society (APS): (since 1969)
       APS Press (the APS book publishing division): 1984-1991 (senior editor: 1984-1987, and
           editor-in-chief: 1987-1991)
       APS Administrative Council: 1987-1991, 1994-1997
       APS Financial Advisory Board: 1987-1991, 1995-1997
       APS representative to International Society of Plant Pathology, 1982-1993.
       APS representative to the Council for Agricultural Science and Technology (1996-1998)
       APS representative for the board responsible for setting Crops and Plant Systems
           Research National Priorities for the 21st Century (CROPS 99).
       APS Pacific Division Councilor: 1994-1997
       Plus service and leadership on many other committees and task forces.
   Australasian Plant Pathology Society: 1986-1989; 1997-present
   International Society for Plant Pathology: since 1982 (Board of Directors, 1982-1993)
   Council for Agric. Science and Technology: since 1984 (Board of Directors; 1995-1998)
   American Society of Agronomy: since 1966
   Western Soil Fungus Conferences: since 1969 (Steering Committee, 1986-1989, 2000-2004;
       Conference Coordinator, 2001, 2005)
   Eastern Research Conferences on Ecology of Root-Infecting Microorganisms: 1975-1985
       (Steering Committee, 1977-1984)
   Society of Nematologists: 2003-present
   Australasian Association of Nematology: 2004-present
   International Turfgrass Society, 1980-1990
   European Network for the Durable Exploitation of Crop Protection Strategies (composed of
       16 federal, state, academic and private institutes in 10 European countries): Board of
       Directors, 2006-present

Honors and Awards:
  Briskey Award for Faculty Excellence, Oregon State University, 1995
  Fellow of the American Phytopathological Society, 1994
  New York Academy of Sciences; 1983

Research Grants at OSU: Approximately $2.5 million since 1986, from the Oregon Wheat
   Commission, USDA Competitive Grants, USDA-Agricultural Research Service Cooperative
   Research Agreements, Rockefeller Foundation, and agrichemical manufacturers.

Publications:                                      Total    Last 10 years Last 5 years
   Refereed technical journal publications:         68         19            12
   Other refereed technical publications:          172         44            30
   Published abstracts:                             47         16            15
   Books, chapters & CDs:                           30         11              2
   Refereed extension bulletins:                     8           4             4
   Non-refereed extension publications:            158         45            38