Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Regional Prairie Native Seed Project
Final Report for CNLM Budget G1005
Sarah Hamman1 Jonathan D. Bakker2 and Sierra Smith1
Prepared for The U.S. Fish and Wildlife Service
US Fish and Wildlife Service
USFWS Agreement # F11AC00065
Webster’s seed farm in full bloom
1
The Center for Natural Lands Management, Olympia, WA 98501; Sarah Hamman - email:
shamman@cnlm.org; phone: 360-790-4180; Sierra Smith - email: ssmith@cnlm.org; phone:
360-480-6105
2
School of Environmental and Forest Sciences, University of Washington, Box 354115, Seattle,
WA 98195-4115; email: jbakker@uw.edu; phone: 206-221-3864
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Summary
Today, native prairies of western Washington are one of the most endangered ecosystems in the
United States. Native prairie habitats have been nearly extirpated from the region and are the
most endangered ecosystem in the state. A number of rare and endangered species are associated
with this habitat, including golden paintbrush (Threatened), Taylor’s checkerspot (Endangered),
Mardon skipper (Species of Concern), and valley silverspot (Species of Concern). The
restoration work conducted through this project occurs across a large portion of these species’
ranges. This project contributes to the recovery of golden paintbrush and these rare butterfly
species. A unique aspect of this project is that it provides crucial restoration information to land
managers while also restoring expanses of critical habitat for rare species on protected prairie
sites. It builds directly upon years of restoration and research, as demonstrated by the strong
support for this project by partners. This project had three components: 1) Seeding technique and
rate assessment, 2) Seed production, coordination and development, and 3) Companion planting
of golden paintbrush. The first component of the project expanded tested seeding rates and
methods for 27 native species at multiple sites. The second component involved increasing
capacity for seed production, the development and documentation of seed handling and
production techniques, and increased regional coordination and networking. The third
component involved an examination of companion planting techniques to improve survival and
performance of outplanted golden paintbrush. We have a high certainty that the benefits of this
project have been realized, and that they will have long-term consequences for prairie habitat
restoration because they contribute to the information base necessary to properly manage these
species.
Background
The prairie ecosystems of Puget Sound contain a disproportionate number of federal and state
listed species, including the following species addressed by this project:
 Taylor’s checkerspot (Euphydryas editha taylori) - Federally Endangered and Washington
State Endangered butterfly species. Recovery guided by the 2009 USFWS Candidate
Assessment and Listing Priority Assignment form.
 Mardon skipper (Polites mardon) - Federal Species of Concern and Washington State
Endangered butterfly species. Recovery guided by the 2009 USFWS Candidate Assessment
and Listing Priority Assignment form.
 Valley silverspot (Speyeria zerene bremnerii) – Federal Species of Concern and Washington
State Candidate butterfly species.
 Island marble (Euchloe ausonides insulanus) – Federal Species of Concern and Washington
State Candidate butterfly species.
 Golden paintbrush (Castilleja levisecta) – Federally Threatened and Washington State
Endangered plant species. Recovery guided by the 2010 Recovery Plan for the Prairie
Species of Western Oregon and Southwestern Washington, 2010 Spotlight Species Action
Plan, 2009 Evaluation of Prairies for Reintroduction in North and South Puget Sound, 2004
Reintroduction Plan, and 2000 Final Federal Recovery Plan.
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
This multi-year project builds on more than a decade of habitat restoration in native prairies and
specifically supports and improves the burgeoning native seed development efforts to restore
habitat for these rare plant and butterfly species in the prairies of western Washington. It
consisted of three components: 1) Seeding technique and rate assessment, 2) Seed production,
coordination and development, and 3) Companion planting of golden paintbrush.
Project Objectives
The objectives of the project were to:
1. Adaptively improve current methods and develop new native seeding techniques for
restoring prairie habitat for rare and endangered butterfly species in Puget Sound.
2. Increase the total production of native prairie seed in the South and North Sound.
3. Support the coordination of regional seed production efforts and the development of a
best management practices (BMPs) handbook and database.
4. Enhance knowledge of BMPs for golden paintbrush recovery and conduct experimental
plantings of golden paintbrush to establish a viable population in at least one new site.
The benefits of this project include:
1. Restoration of 25 acres of habitat for Taylor’s Checkerspot and other rare butterflies in
South Puget Sound;
2. Development of BMPs for seed collection, storage, and quality control.
3. Large-scale seed production of over 100 native species and seed purity information on
over 50 species.
4. Testing and development of seeding techniques and field germination estimates at large
scales.
5. Experimental plantings of golden paintbrush to establish one new population with 1000
flowering individuals in the South Sound and to augment an existing population in the
North Sound.
Project Details
This report summarizes the work that has been completed during this multi-year project for each
of the three focus areas: (1) Seeding technique and rate assessment, (2) Seed production,
coordination and development, and (3) Companion planting of golden paintbrush. Results from
prior years were reported in Hamman et al. (2010, 2011, 2012, 2013, 2014).
(1) Seeding Technique and Rate Assessment
Phases I-II: The Seeding Technique and Rate Assessment study was replicated at four sites in
the South Sound during Phase I (2009-2010): Glacial Heritage, Tenalquot, Scatter Creek, and
JBLM. In Phase II (2010-2011), we replicated the entire project at all but one (JBLM) of the
original sites. We were unable to continue the project at JBLM due to disruption of the plots by
off-road training and off-road
At each site, research plots were located in areas that had been prepared for larger scale seeding
through restoration and enhancement efforts over previous years (prescribed fire, mowing and
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
herbicide treatments); thus building on a foundation of successful restoration work. We tested
three seeding techniques (seed drill, broadcast seeder, hydro-seeder), and five seeding rates (0,
350, 700, 1050, 1400 seeds/m2). Because seed weights differ among species, we sowed on the
basis of seed density rather than combined weight. With the mixture of species, these rates were
equivalent to approximately 0, 2, 4, 6, and 8 lbs per acre, respectively. In Winter 2009, we used
Festuca roemeri, Eriophyllum lanatum and Castilleja hispida with a grass:forb ratio of 1:1.3,
except at McChord, where only F. roemeri was used due to site conditions. In Fall 2010, we used
four species: Festuca roemeri, Eriophyllum lanatum, Castilleja hispida and Potentilla gracilis
with a grass:forb ratio of 1:1.
The experiment had a 3x5 randomized factorial block design with three replicate blocks per siteyear and no replication within blocks. The experimental units were strips, sized such that
operationally realistic equipment could be used. The total research areas varied slightly in size
between sites, but were all approximately the same size for Phases I-II (Table 1). Overall
research area was much smaller in Phases III-IV due to reduced plot size and fewer treatment
plots.
Table 1. Research area details (acres) for each site throughout the entire project
Site
2009
2010
2012
2013
Glacial Heritage Preserve
Tenalquot Preserve
Scatter Creek Wildlife Area
JBLM
Total Research Area
1.94
1.40
0.90
1.85
6.09
1.80
1.58
1.22
N/A
4.60
0.16
0.16
N/A
N/A
0.32
0.16
N/A
N/A
0.16
0.32
The strips were seeded using either a Kasco no-till seed drill, a Truax broadcast seeder or a
hydroseeder (Figure 1). Because the hydroseeder was designed to sow seeds at extremely high
rates (10-30lbs/acre), we had to adjust our methodology to sow native seeds using lower rates.
To simulate the hydroseeding treatment, we applied the seeds using the broadcast seeder and
then sprayed Hydrostraw mulch (at the standard 2000 lbs/acre) over the seeds evenly across the
treatment strips. In 2010 - 2013, we monitored seedling establishment in the experimental plots.
We established five 1 x 1-m quadrats along the central transect of each strip. Distance between
quadrats ranged from 2m to 7m, depending on the size of the strip. We counted the total number
of individuals of each seeded species within each quadrat in June of each year following seeding.
Older established individuals were not separated out from new seedlings because we assumed
this distinction would be impossible to make within two or three years of seeding. C. hispida was
so infrequent in plots that only frequency instead of density was recorded in 2011 and 2012. In
the third year of monitoring, due to the rhizomatous spread of E. lanatum, we estimated percent
cover rather than number of individuals. Percent cover was a visual estimate of foliar cover
within cover bins (0%, 0-1%, 1-5%, 6-10%, 11-20%, 21-30%, 31-50%, 51-70% 71-90%, 91100%). For the purposes of analysis, the median of the bin was used as the cover estimate for
each quadrat. Percent bare ground was also recorded in 2011.
We analyzed comparisons of seeding methods and rates using meta-analysis and moderated
multiple regression procedures outlined in Viechtbauer (2010), Stewart et al. (2008), Harrison
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
(2011). Each site and year sown combination was treated as a separate study (n = 6). We
transformed the density (plants/m²) and percent cover data using ln(y+1) to normalize the
distribution. We examined two different effect sizes: standardized mean difference Hedges’s g
(Hedges 1981) for method treatment effects, and Pearson’s correlations transformed into a
Fisher’s z for rate treatment effects (Fisher 1958; Corey et al. 1998). Table 2 displays a summary
of which effect sizes were used for exploring relationships between treatment and outcome
variables.
Figure 1. Treatment strips were seeded using a seed drill (a), broadcast seeder (b) or hydroseeder (c).
Since the aim of the study was to determine sources of variation in sown species occurrence
related to both experimental treatment effects (method and rate) and uncontrolled factors (site
and month sown), we ran mixed effects regression (ME) models with moderators, followed by
separate random effects (RE) models for subsets of any moderators that explained a significant
proportion of variance in the full model. We ran the meta-regression models in R Studio 3.1 (R
Studio 2010), using the package metafor, with a restricted maximum likelihood estimator
(Viechtbauer 2010). Calculations for the REML estimator for mean and variance are described
by Viechtbauer (2005). The associations between different sowing method (n = 3) required three
different models for multiple comparisons, with site and month sown specified as moderators.
The association between rate and the dependent variables only required one model, with method,
site, and month sown specified as moderators. We created Forrest plots showing estimated
treatment effect sizes from each study, along with overall RE model effect size or effect sizes, if
moderators produced significant variability. When applicable, we back-transformed data for
presentation purposes.
No sowing method performed significantly better than any others under all conditions. Although
seed drilling was most effective at establishing sown species in Winter 2009 plots, broadcasting
produced more reliable results overall (Figure 2). Besides producing high first year densities of
Potentilla gracilis in Fall 2010 plots, hydroseeding was regularly the second or third best
method. Although significant effect sizes for method treatments were relatively small (0.21 to
0.42), the impact of an additional 5 – 10 sown plants/m² across a large-scale restoration project
could be substantial. Contrary to expectations, the best sowing method was not site specific;
differences between methods were relatively constant across sites.
Overall, sown plant densities at Glacial and Scatter Creek were higher for plots sown in Winter
2009 than those sown in Fall 2010. Tenalquot sown plant densities were higher in Fall 2010 plots
(Fig. 2). Individual sown species also displayed lower densities in Fall 2010 seed drilled plots
5
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
compared to broadcasting and hydroseeding (Table 2). Hydroseeding did a poor job of
establishing Festuca roemeri in Winter 2009 plots, but was not significantly different from any
other method in Fall 2010 plots (Table 2). Method had no effect on frequency of Castilleja
hispida occurrence (ME model of pooled log odds ratio between methods, p > 0.05).
Fall 2010
Winter 2009
Figure 2. Individual site and month effect sizes comparing three methods of sowing: a) Broadcast (BC) vs. Seed drill
(SD), b) Broadcast (BC) vs. Hydroseed (HS), and c) Seed drill (SD) vs. Hydroseed (HS). The squares represent mean
effect size of individual studies with 95% Confidence Intervals. Diamonds represent pooled effect sizes generated
using standardized mean difference random effects meta-analysis (95% CI is indicated by diamond width). Densities
are back-transformed. C. hispida was not included in total sown plant density.
Across all sites, methods, and years sown, sowing rate was positively correlated with first year
total sown species density (plants/m²) (Fig. 3). Year sown explained 35% of between-group
variance, while method and site collectively accounted for less than 10% of between-group
variance. The correlation between sowing rate and total sown species density was stronger for
plots sown in Winter 2009, as opposed to those sown in Fall 2010 (Fig. 3). The strength of the
relationship between rate and sown species density was moderate (r² = 0.46 for Winter 2009 and
0.26 for Fall 2010), indicating that other unexplained variables influence sown plant densities.
The average target density (14 plants/m²) for F. roemeri was exceeded by the highest sowing rate
of 840 seeds/m² but was not met by the second highest rate of 630 seeds/m² (Fig. 3a). The
average target density (0.2 plants/m²) for Potentilla gracilis was exceeded by the lowest sowing
rate tested of 70 seeds/m². Even by the third year, C. hispida was only present in approximately
12% of sown plots (Fig. 3c). Average density of C. hispida for all sowing rates tested was less
than 0.5 plant/m², well below the desired 4 plants/m² habitat target.
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Fall 2010
Winter 2009
Figure 3: Effect sizes of the relationship between sowing rate and sown species density for each site, month, and
method. The squares represent mean effect size of individual ‘studies’ with 95% Confidence Intervals. Diamonds
represent pooled effect sizes generated from Fisher’s z random effects meta-analysis (95% CI is indicated by
diamond width). C. hispida was not included in total sown species densities.
As expected, higher seeding rates usually resulted in higher plant density. In this experiment, we
could not measure exact germination rates, as there were low background levels of all sown
species except C. hispida. However, the density of all sown species never topped 10% of the
amount of sown seed and was often less than 5%, indicating that germination was very low. At
the time of sowing for this experiment, our seed production operation did not take measurements
of seed viability, so all of our sowing rates were calculated in terms of bulk seed and not pure
live seed (PLS). Recent (2013) PLS estimates for the species used in this experiment were 12%
for F. roemeri, 52% for P. gracilis, and 79% for Eriophyllum lanatum (Hamman et al. 2013).
Now that seed production has evolved in the South Sound Prairies and seed cleaning procedures
are improved, we have measures of seed viability and are staged to increase field germination.
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
a
b
c
d
Figure 4. Linear regressions of the relationship between sowing rate (seeds/m2) and third year density or cover:
a) F. roemeri, b) E. lanatum, c) P. gracilis, d) C. hispida. Larger points indicate multiple observations at a
particular data value. 95% confidence intervals for the mean density are shown as a gray band around the
regression line. Density (plants/m2) and cover (%) have been transformed but the y axis is labeled in the original
units for ease of interpretation. Figures a, c, and d have horizontal lines indicating butterfly habitat targets
Seed predation also likely led to low field establishment rates. While burning our sites prior to
sowing was necessary to create space for seed-soil contact, litter removal makes seeds more
available to rodents. Future work could attempt to quantify levels of seed predation in South
Puget Sound prairies.
Results from this project have been presented at annual conferences of the Northwest Scientific
Association, Ecological Society of America and the National Native Seed Conference. Results
are currently being written up for publication in Restoration Ecology.
Phases III-IV: In Phases III (2012) and IV (2013) we utilized some of the lessons learned in
Phases I and II and installed research plots that tested fewer metrics on more species. At each site
(2012: Glacial & Tenalquot; 2013: Glacial & South Weir) research plots were located in areas
that had been prepared for larger scale seeding through restoration and enhancement efforts, thus
building on a foundation of successful restoration work. We tested effects of two seeding
techniques (seed drill, broadcast seeder), and three seeding rates (control, low, high) on
establishment of 27 species (Table 3). Because seed weights differ among species, we sowed on
the basis of seed density rather than combined weight. We altered seeding rates for several
species in 2013 based on PLS estimates found by J. Bakker’s lab (see ‘Seed germination and
viability testing’ below) and initial germination rates measured in the 2012 seeded plots. We also
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
added several annual species in 2013, as sufficient seed was available and managers requested
information on seeding rates for these important butterfly resource species.
Table 2. Species and rates (bulk seed) used in 2012 and 2013 research plots. Seeding rates are presented as number
of seeds per square meter.
Species
Achillea millefolium
Armeria maritima
Balsamorhiza deltoidea
Carex inops
Cerastium arvense
Clarkia amoena
Collinsia grandiflora
Collinsia parviflora
Danthonia californica
Eriophyllum lanatum
Erigeron speciosus
Festuca roemeri
Koeleria macrantha
Lomatium triternatum
Lomatium utriculatum
Lupinus albicaulis
Lupinus bicolor
Lupinus lepidus
Microseris laciniata
Plectritis congesta
Potentilla gracilis
Ranunculus occidentalis
Sericocarpus rigidus
Sisyrinchium idahoense
Solidago missouriensis
Solidago simplex
Viola adunca
Codon
ACMI
ARMA
BADE
CAIN
CEAR
CLAM
COGR
COPA
DACA
ERLA
ERSP
FERO
KOMA
LOTR
LOUT
LUAL
LUBI
LULE
MILA
PLCO
POGR
RAOC
SERI
SIID
SOMI
SOSI
VIAD
2012
Annual /
Perennial
LOW
P
P
P
P
P
A
A
A
P
P
P
P
P
P
P
P
P
P
P
A
P
P
P
P
P
P
P
6
4
0
12
12
12
0
0
15
3
4
28
8
10
12
2
6
6
9
0
6
12
70
8
12
12
18
2013
HIGH
21
14
0
42
42
42
0
0
53
11
14
98
28
35
42
7
21
21
32
0
21
42
245
28
42
42
63
LOW
5
7
5
0
9
24
12
16
10
3
5
78
7
20
24
4
12
0
18
16
21
8
156
16
24
24
36
HIGH
18
25
18
0
32
84
42
56
35
11
18
273
25
70
84
14
42
0
63
56
74
28
546
56
84
84
126
As before, the experiment had a randomized block design with three replicate blocks per siteyear and no replication within blocks. The experimental units were strips, sized such that
operationally realistic equipment could be used (approximately 36m2). The strips were seeded
using either a Kasco no-till seed drill or a Truax broadcast seeder in late October or early
November of each year.
9
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Three 1m x 1m quadrats were established evenly along a central transect within each strip for
vegetation monitoring. First and second year monitoring was conducted in the middle of the
flowering season (late May/early June) and included the following variables:
 Number of germinants (abundance) of each seeded species
 Percent cover of bare ground in each quadrat
 Percent cover of moss and lichen
These results confirm
that seeding methods do
not influence
establishment in any
% estab. (seedlings/# seeds sown)
% estab. (seedlings/# seeds sown)
Overall findings from Year 1 and 2 establishment in the 2012 plots show that establishment rates
ranged from 0% to 33% across species and sites (Figure 5). Only ten of the 24 species sown in
2012 had densities above the unseeded control plots, therefore results for these ten species are
presented. Again, seeding method was not significant for most species, however site was
significant for nearly all
35
species. Seeding rate was
a
significant for only seven
Glacial Heritage
30
species across the two sites
Year 2
(Figures 6a and b). The
25
Year 1
‘high’ seeding rate was
20
effective at establishing
more than one plant per
15
square meter for all
species, while the ‘low’
10
seeding rate only achieved
5
this target for three species
(C. amoena, M. laciniata,
0
S. rigidus). S. rigidus
CEAR CLAM ERSP LOTR LUAL LUBI POGR SERI SIID SOSI
establishment was
extremely high in year
b 30
one, despite estimates of
Tenalquot
25
extremely low PLS for
this species (Appendix
20
2). Germination was
positively correlated with
15
bare ground and for four
species (C. amoena, L.
10
bicolor, D. californica,
M. laciniata).
5
0
CEAR CLAM ERSP LOTR LOUT LUAL POGR SIID SOMI VIAD
Figure 5. Range (low to high) of percent establishment for 10 seeded species, both
one and two years post-seeding at a) Glacial Heritage and b) Tenalquot.
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
consistent pattern. Therefore, the most cost effective seeding method (broadcast seeding) should
be used to maximize efficiency. Additionally, optimal seeding rates vary by species and site.
Finally, sufficient bare ground must exist at restoration sites prior to seeding to ensure successful
native establishment of all seeded species.
a
b
Figure 6.
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Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
(2) Seed Production, Coordination and Development
Seed Production
The South Sound Conservation Nursery
program now utilizes multiple seed farms in
Thurston and Pierce counties to produce large
lots of core restoration species (Figure 7; Table
3). The seed farm at the Violet Prairie Scatter
Creek preserve has become our primary
production site and the hub of the seed
production program. In 2014 we expanded from
Figure 1. 2014 bloom at Webster’s Nursery
20 rows to 120 rows, which filled our first five
acres. This massive planting effort involved
over 100,000 transplants and substantial direct
seeding. We also converted an additional five
acres of pasture at Violet Prairie to fully
Figure 7. Oregon sunshine at Violet Prairie Seed
irrigated row production that will accept seeds
Farm
and transplants in the spring of 2015. Rare and
specialty restoration species are produced in 62
Table 3: 2014 Regional Seed Production Summary
raised seeds beds at Shotwell’s Landing Nursery
Pounds
Number
in Rochester, WA (Figure 8).
Source
Produced of Lots
The program also contracts with large growers for
Webster’s Seed Farm
459.75
60
Roemer’s fescue production.
Violet Prairie Seed Farm
The South Sound seed production program grew
aggressively in 2014, and provided a total of 2109
pounds of seed to South Sound land managers in
2014 (Table 3), a 152% increase over 2013. Seed
production from CNLM farm sites increased by
61% to 690 pounds (Table 4). The new Violet
Prairie farm site is averaging an encouraging three
times the production by area as our historic
Webster’s seed farm. As we transition away from
Webster’s and begin production in earnest on better
soils, we expect the upward trajectory in production to
continue.
Finally, the nursery program maintains a central seed
processing and storage facility to increase efficiency
and tracking of over 100 species for all partner
agencies (Figure 9). This facility has expanded
dramatically over the past three years to
accommodate the ever-increasing volume of seed.
A database has been developed and maintained to
track production and protocols for all species.
12
Shotwell’s Landing
Cavness Ranch
144.74
16
44.39
46
41.60
1
1368.00
2
West Rocky Seed Beds
0.70
3
Bald Hills Seed Beds
1.13
5
48.80
2109.11
149
282
Fescue Contracts
Wild Collection
TOTAL 2014 Seed
Figure 8. Rare species Royal Jacob’s ladder
at Shotwell’s Landing Nursery
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Table 4. Seed production at CNLM seed farms over past three years.
Species
Achillea millefolium
Allium amplectens
Aquilega formosa
Arabis glabra
Armeria maritima
Balsamorizha deltoidia
Bromus pacificus
Camassia leichlinii
Camassia quamash
Campanula rotundifolium
Carex inops
Carex tumulicola
Castilleja hispida
Castilleja levisecta
Castilleja miniata
Cerastium arevense
Cirsium brevistyllum
Clarkia amoena
Collinsia grandiflora
Collinsia parviflora
Collomia grandiflora
Danthonia californica
Danthonia spicata
Delphinium nutallii
Dichanthelium acuminatum
Dichanthelium oligosanthes
Dodecatheon hendersoni
Dodecatheon pulchellum
Drymocallis glandulosa
Elymus glaucus
Elymus trachycaulus
Erigeron specious
Erigeron strigosus
Eriophyllum lanatum
2014 seed
production
(lbs)
4.088
0.278
1.523
0.49
21.10
13.33
1.39
1.25
14.83
0.47
0.02
0.10
13.92
15.70
0.01
1.83
0.00
7.21
38.47
45.37
8.37
13.59
0.42
0.24
0.31
0.27
0.04
0.03
3.97
2.49
0.53
11.78
0.03
80.82
2014 bulb
production
(lbs)
10.28
17.1
13
2013
Produciton
(lbs)
3.59
0.15
1.61
0.00
15.26
6.69
2012
Production
(lbs)
1.91
0.08
0.27
0.00
19.48
0.27
0.55
0.32
1.10
0.06
0.00
0.05
0.17
0.13
4.86
2.11
2.66
1.17
2.39
0.02
1.98
13.89
57.85
1.41
3.29
0.92
0.87
0.80
0.10
10.20
0.80
3.14
0.00
28.21
0.06
0.34
0.01
0.00
4.10
1.84
3.21
3.24
3.18
2.53
49.34
18.39
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Table 4. cont’d.
Species
Festuca roemeri
Fritillaria affinis
Gaillardia aristata
Gilia capitata
Heuchera chlorantha
Hieracium cynaglossoides
Iris tenax
Koeleria macrantha
Leptosiphon bicolor
Ligusticum apiifolium
Lithophragma parviflorum
Lomatium nudicaule
Lomatium triternatum
Lomatium utriculatum
Lotus nevadensis
Lupinus albicaulis
Lupinus bicolor
Lupinus lepidus
Lupinus polyphyllus
Luzula comosa
Micranthes integrifolia
Microseris laciniata
Microsteris gracilis
Naveretta intertexta
Navarretia squarrosa
Nuttallanthus canadensis
Perideridia gairdneri
Plantago lancelota
Plectritis congesta
Poa secunda
Polemonium carneum
Potentilla gracilis
Ranunculus occidentalis
Ranunculus orthorhynchus
2014 seed 2014 bulb
2013
production production Produciton
(lbs)
(lbs)
(lbs)
1452.45
313.41
0.02
11.71
1.42
23.96
0.98
1.10
1.33
0.09
0.03
1.76
0.00
11.57
3.06
2.18
0.00
0.71
0.05
0.14
0.02
1.50
0.09
12.80
9.70
5.70
7.00
0.01
39.97
22.71
36.04
20.14
2.90
0.41
0.30
0.13
0.21
0.10
9.53
7.38
11.32
12.33
0.08
1.24
2.95
1.33
1.23
0.57
0.47
0.00
48.28
13.42
39.88
56.20
0.01
0.01
18.36
3.55
26.31
5.64
0.05
0.03
14
2012
Production
(lbs)
427.58
0.77
0.00
1.08
0.00
0.00
12.59
0.00
0.05
0.05
0.05
8.17
1.96
21.60
10.26
4.42
0.13
5.26
1.39
0.00
0.00
0.03
0.29
1.83
0.92
2.89
3.44
0.12
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Table 4. cont’d.
Species
Rhinanthus minor
Rupertia physoides
Sanicula crassicaulis
Sanicula graveolens
Sericocarpus rigidus
Sidalcea nelsonianna
Sidalcea virgata
Silene douglasi
Silene scouleri
Sisyrinchium idahoense
Solidago missouriensis
Solidago missouriensis 'Glacial'
Solidago simplex
Symphyotrichum hallii
Synthyris reniformis
Toxicoscordion venenosus
Trifolium wildenovii
Trillium parviflorum
Triodanis perfoliata
Triteleia hyacinthina
Vicia americana
Viola adunca
Viola howelii
Viola praemorsa
Wyethia angustifolia
Totals
2014 seed 2014 bulb
2013
2012
production production Produciton Production
(lbs)
(lbs)
(lbs)
(lbs)
0.14
0.02
0.11
0.07
0.00
0.00
0.02
0.01
3.33
2.69
2.05
3.14
1.61
1.43
0.11
6.92
0.08
0.11
0.26
0.08
0.02
7.72
4.23
5.85
4.31
1.12
6.84
0.62
0.09
0.21
2.53
2.93
2.07
0.66
0.45
2.69
0.01
0.08
0.02
0.00
4.00
1.97
0.63
0.97
0.24
0.00
0.83
0.93
0.00
0.03
0.00
0.00
0.07
3.09
2.11
2.66
0.03
1.62
0.47
0.07
0.58
0.01
0.00
2109.12
27.38
678.61
626.85
Seed germination and viability testing
Seed germination and viability testing occurred in
the Terrestrial Restoration Ecology lab at the
University of Washington.
Testing occurred for 65 accessions from 57
prairie species (Table 5). Our general testing
procedure was as follows: We separated each
accession into pure seed and debris (chaff, seed of
other species, etc.), and weighed each component.
We also counted the seeds and calculated the
15
Figure 9. Seed cleaning center
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
bulk density (seeds per gram of material, including debris). We calculated purity as seed mass
divided by total mass of sample. We then reviewed the literature and determined the most
appropriate of three germination regimes (spring/fall, summer, winter) for each species.
Environmental conditions in each regime were as follows:
 Spring/Fall: 15°C and light for 12 hours; 8°C and dark for 12 hours
 Summer: 24°C and light for 14 hours; 14°C and dark for 10 hours
 Winter: 5°C and light for 10 hours; 2°C and dark for 14 hours
We counted out 400 seeds per accession and divided them into 4-8 lots. Each lot of seed was
placed on filter paper in a petri dish in the growth chamber. The filter paper was kept moistened
during testing. Each lot was checked periodically, and germinants were counted and removed.
At the end of the germination period, non-germinating seeds were subject to tetrazolium testing
to identify those seeds that remained viable (e.g., are dormant). We calculated the percentage of
seeds that germinated and the percentage of seeds that were viable; these two percentages sum to
the percentage live seed. We calculated the percent dormant seed as the number of dormant
seeds divided by the total number of live seeds. The percent pure live seed (PLS) was
calculated as the product of purity and live seed.
Through this testing, we have identified accessions with particularly low PLS values due to poor
seed purity and/or low percentages of live seed. These accessions would have to be seeded at
much higher rates than other species to compensate for the low PLS values. Notably,
Dodecatheon hendersonii, Koeleria macrantha, Ligusticum apiifolium, Lithophragma
parviflorum, Zigadenus venosus and Viola adunca had < 20% PLS and Navarretia intertexta,
Sericocarpus rigidus and Solidago missouriensis had < 10% PLS (Table 5), meaning that land
managers seeding them at regular rates would only be putting out one fifth to one tenth the
amount of seed needed to achieve target densities. We have also learned how to improve
germination through experimental testing of stratification methods. For example, germination of
Navarretia intertexta and of Lupinus spp. was low until we began to prick each seed to enhance
water movement through the seedcoat. This process also allowed us to identify species that
require additional study. For example, our testing regime did not break dormancy for any
Ligusticum apiifolium seeds and for only 1% of the live Viola praemorsa seeds. Testing with
longer stratification times and/or other regimes is clearly necessary for these species.
16
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Table 5. Germination and viability data for species and accessions tested at the University of Washington during the reporting period. Species
abbreviations represent the first two letters of the genus and first two letters of the species. Red highlight shows species with PLS < 20%.
Abbrev.
UW #
CNLM #
N
Testing Regime
Bulk
Purity
Live Seed
Dormant
PLS
Density
Seed
(seeds/g)
(%)
(%)
(%)
(%)
ACMI
ALAM
ALAM
AQFO
ARMA
BADE
CAHI
CAIN
CALE
CAQU
CARO
CEAR
CIBR
CLAM
COGR
COGRX
COPA
DACA
DASP
DENU
DENU
DOHE
ELGL
ELGL
ERLA
ERSP
FERO
GAAR
HECH
222
223
285
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
286
241
242
283
243
244
245
246
247
2246
2128
2128
2145
2207
2203
2196
2136
2121
2092
2263
2103
2125
2123
2108
2216
2104
2220
2202
2159
2159
2097
2221
2221
2211
2214
2127
2247
2199
400
200
183
400
400
400
400
400
400
400
400
400
400
400
400
400
387
400
200
200
400
199
192
400
400
272
400
399
Summer
Winter, Spring, Winter
Winter, Spring, Winter, Spring, Winter
165 d Winter, then Spring
Spring
42 d Winter, then Summer
52 d Winter, then Summer
Summer
84 d Winter, then Spring
80 d Winter, then Spring
31 d Winter, then Summer
Summer
Spring
Spring
Spring
Spring; poked
Spring
Summer
30 d Winter, then Spring
Winter + bagged (dark)
Winter; light
84 d Winter, then Spring
15 d Winter, then Summer
Summer
20 d Winter, then Spring
34 d Winter, then Summer
Spring
Spring
68 d Winter, then Spring
17
3922
357
357
509
352
106
6012
327
114
227
11478
6452
692
2657
1466
280
741
231
643
1065
1065
967
247
247
2120
3497
178
232
16059
41.2
98.2
98.2
99.4
86.7
92.0
78.1
99.9
99.0
99.5
97.6
100.0
90.8
80.4
96.4
99.2
95.3
89.3
97.4
93.7
93.7
98.9
87.5
87.5
92.8
94.4
56.3
64.2
96.4
87.5
66.0
88.5
79.3
79.8
38.8
59.5
66.3
53.8
81.8
67.5
92.8
33.0
85.5
81.8
80.0
86.3
82.7
90.8
76.5
77.0
10.8
85.9
88.5
78.3
81.3
61.4
51.5
52.9
2.0
50.8
4.9
89.9
3.4
63.9
18.5
67.2
48.8
9.5
4.8
2.7
32.6
1.5
0.9
20.9
25.8
15.6
1.7
8.5
24.7
20.9
1.2
0.0
7.0
0.6
3.0
68.0
2.6
36.1
64.8
86.9
78.8
69.1
35.7
46.5
66.2
53.2
81.3
65.9
92.8
30.0
68.7
78.8
79.4
82.2
73.8
88.4
71.7
72.1
10.6
75.2
77.4
72.6
76.7
34.6
33.1
51.0
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Abbrev.
UW #
CNLM #
N
Testing Regime
Bulk
Density
(seeds/g)
Purity
(%)
HICY
HICY
KOMA
LIAP
LIPA
LONU
LOTR
LOUT
LUAL
LUAL
LUBI
LULE
MIGR
MIIN
MILA
248
289
249
250
251
252
253
254
255
256
257
258
259
260
261
2153
2153
2210
2149
2115
2152
2206
2205
2212
2212
2110
2242
2111
2096
1838
200
200
400
435
409
400
400
400
396
400
387
400
400
400
400
2004
2004
4908
316
181818
122
122
443
29
32
156
219
738
12300
654
96.2
96.2
83.4
98.4
90.9
93.2
94.6
91.5
100.0
99.8
73.6
97.0
92.6
95.3
96.0
NAIN
NAIN
NASQ
NUCA
PLCO
PLLA
POGL
POGR
RAOC
SERI
SIDO
SIID
SINE
SISC
SOMI
262
287
263
264
265
266
267
268
269
270
271
272
273
274
275
2243
2243
2219
2116
2105
2241
2201
2197
2100
2244
2217
2198
2200
2264
2253
109
318
378
400
400
400
400
400
400
397
400
400
400
400
400
Winter, then Spring
Spring
Summer
30 d Winter, then Spring or Summer
Spring
56 d Winter, then Spring
82 d Winter, then Summer
26 d Winter, then Spring
Poked; Summer
Poked; Summer
Scarified (hot water, razor); Summer
Hot water, nicked; Summer
14 d Winter, then Spring
15 d Winter, then Spring
Summer
53 d Spring, 36 d Winter, 11 d Spring,
35 d Winter, Spring; poked near end of
test
Various
Spring
Winter then Summer
Winter
Summer
63 d Winter, then Summer
34 d Winter, then Spring
38 d Winter, then Summer
43 d Winter, then Summer
56 d Winter, then Spring
63 d Winter, then Summer
Scarified (jar w/ rocks); Spring
56 d Winter, then Spring
Summer
2772
2772
15738
44444
822
442
2881
2633
436
551
1259
581
293
1349
8937
97.0
97.0
94.4
100.0
66.1
74.5
98.7
87.5
99.3
15.6
99.8
100.0
96.2
77.9
49.9
18
Live Seed
PLS
(%)
Dormant
Seed
(%)
68.5
78.5
14.8
16.6
93.9
20.8
83.5
90.8
27.5
35.3
84.2
66.3
69.5
92.8
72.0
27.0
5.1
0.0
100.0
0.5
50.6
4.8
5.2
9.2
0.0
0.0
29.1
1.8
1.9
12.8
65.9
75.5
12.3
16.3
85.3
19.3
79.0
83.0
27.5
35.2
62.0
64.3
64.4
88.4
69.1
27.5
6.9
75.7
49.8
43.8
76.5
74.8
58.0
48.3
22.9
28.5
28.3
68.3
57.5
8.3
20.0
86.4
1.0
3.5
8.6
40.2
8.7
5.6
18.1
11.0
9.6
29.2
24.2
0.9
30.3
26.7
6.7
71.4
49.8
28.9
57.0
73.8
50.8
48.0
3.6
28.4
28.3
65.7
44.8
4.1
(%)
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Abbrev.
UW #
CNLM #
N
Testing Regime
Bulk
Density
(seeds/g)
Purity
(%)
SOSI
SOSI
TOVE
276
277
278
2222
2252
2180
400
400
406
1958
2565
561
71.0
78.9
99.8
TRWI
VIAD
VIPR
279
280
281
2117
2224
2189
399
395
400
Summer
Summer
42 d Winter, then Spring
Scarified (hot water, razor); 33 d
Winter, then Spring
87 d Winter, then Spring
93 d Winter, then Spring
755
1859
240
90.8
99.5
99.8
19
Live Seed
PLS
(%)
Dormant
Seed
(%)
79.3
83.3
12.3
1.3
1.2
16.0
56.3
65.7
12.3
76.4
9.4
25.8
0.0
43.2
99.0
69.4
9.3
25.7
(%)
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
(3) Companion Planting of Golden Paintbrush
This component of the project was conducted at Glacial Heritage Preserve in South Sound. The
purpose of this component of the project was to help determine whether growing golden
paintbrush (CALE) with companion plants for it to parasitize provides significant benefits in the
establishment and maintenance of recovery populations of golden paintbrush in the region. Two
primary experiments were conducted.
Experiment One: Two factors were tested in a factorial design:
pot size (4” pot or 1” tube) and host plant identity (none,
Eriophyllum lanatum, and Festuca roemeri). The CALE were
grown without host plants in the nursery. Host plants were
grown in 1” tubes in the nursery, and outplanted immediately
adjacent to the CALE to maximize the potential for CALE to
establish haustorial connections with them afterwards.
Outplantings occurred in a field at Glacial Heritage. Site
preparation in planted areas consisted of spraying glyphosate
and tilling. To assess the generality of the results, the
experiment was repeated in two years, 2010 and 2011. 576
CALE plants were installed in November 2010 (80-100 per
treatment), and 594 CALE plants were installed in November
2011 (97-100 per treatment). Each plant was individually
labeled so that its performance could be tracked. Plants were
weeded periodically to maintain differences in host identity.
During this reporting period, we continued to maintain these
experimental areas. In Fall 2013, we monitored the number of
Figure 10. CALE plants
fruiting stems and number of capsules of each surviving CALE
grown with F. roemeri (above)
plant in both arrays. In Spring 2014, we monitored survival and
and E. lanatum (below).
vigor of each surviving plant in both arrays, and the number of
flowering stems and maximum stem length of each surviving
CALE plant in the 2011 array. In Fall 2014, we monitored the number of fruiting stems and
number of capsules of each surviving CALE plant in the 2011 array.
Experiment Two: In Fall 2012, Natalie Footen (graduate student) initiated a new experiment to
test the performance of CALE when grown with a wider array of host plants. Plants were grown
in 4” pots in the greenhouse and outplanted into a field that had received several years of
periodic glyphosate applications to control invasive weeds. Twenty CALE plants were planted
with each of 11 host plant species: Achillea millefolium, Danthonia californica, Deschampsia
caespitosa, Eriophyllum lanatum, Erigeron speciosus, Festuca roemeri, Lupinus lepidus,
Lupinus littoralis, Rosa nutkana, Solidago canadensis, and Symphoricarpos albus. A no-host
treatment was also included, but rather than planting a single CALE with no competition, two
CALE plants were planted together so that competition was still occurring. We monitored
survival, vigor, number of flowering stems, and maximum stem length in Spring 2014, and the
number of fruiting stems and number of capsules of each surviving CALE plant in Fall 2014.
Results from Experiment One indicate that the presence of a host plant continues to improve
survival, and that host identity matters: performance of golden paintbrush is enhanced more by
20
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Eriophyllum lanatum than by Festuca roemeri (Delvin 2013). Data from 2014 have not yet been
entered, but our observations are that this effect is persisting beyond 2013 (Figure 11).
Preliminary results from Experiment Two indicated 55% survival of Castilleja levisecta over the
first winter. Surviving plants were much smaller than those in Experiment One, perhaps due to
nursery cultural practices (e.g., container type, lack of fertilization). Survival ranged from a low
of 15% when planted with Lupinus lepidus to a high of 85% when planted with Deschampsia
caespitosa. Results have been presented at two events: Invited lecture at the University of
Maryland, College Park (February 2014) and Presentation at the University of Washington
Undergraduate Research Symposium (May 2014).
2011 Array
2010 Array
Figure 11.
1. Survival of Castilleja levisecta planted alone (No host), with Eriophyllum lanatum, or with
Figure
Festuca roemeri in Fall 2010 and Fall 2011. Line color and symbol shape distinguish host identity
treatments; line type (solid or dashed) and symbol shading (filled or hollow) indicate pot size treatments.
21
Regional Prairie Native Seed Project – Final Report for CNLM Budget G1005
Literature Cited
Delvin, E.G. 2013. Restoring abandoned agricultural lands in Puget lowland prairies: a new
approach. Dissertation. University of Washington, Seattle, WA.
Hamman, S., E. Delvin, and J.D. Bakker. 2010. Regional prairie native seed project: first annual
report. Prepared for US Fish and Wildlife Service, Washington Fish and Wildlife Office,
Lacey, WA. 22 p.
Hamman, S., E. Delvin, and J.D. Bakker. 2011. Regional prairie native seed project: second
annual report. Prepared for US Fish and Wildlife Service, Washington Fish and Wildlife
Office, Lacey, WA. 15 p.
Hamman, S., E. Delvin, and J.D. Bakker. 2012. Regional prairie native seed project: third
annual report. Prepared for US Fish and Wildlife Service, Washington Fish and Wildlife
Office, Lacey, WA. 16 p.
Hamman, S., J.D. Bakker and S. Smith. 2013. Regional prairie native seed project: Final Report
for The Nature Conservancy. Prepared for The Nature Conservancy, Seattle, WA and US.
Fish and Wildlife Service, Washington Fish and Wildlife Office, Lacey, WA. 15.
22