Progress 01/01/12 to 09/30/16
Outputs Target Audience:The target audience reached by this project included US Forest Service Personnel at the Manitou Springs Experimental Forest, colleagues at the New Mexico State University John T. Harrington Forestry Research Center, and staff at Mountain Cement in Laramie and Boulder County Parks and Open Space near Boulder, Colorado. These were formal experiential, outreach and extension opportunities including collaborative efforts between personnel at the University of Wyoming, New Mexico State University and Manitou Springs Experimental Forest. We also spoke with attendees at the Southern Rockies Seed Network in Fort Collins, Colorado and so reached a larger audience of federal agency personnel, local and state employees, and interested parties who work with non-profit organizations. Informal discussions through Extension included landowners, practitioners and industry professionals in the field of ecological restoration. Changes/Problems:Major changes include the reduction in effective number of four sites to three sites for the common garden study due to high rates of seedling mortality at one location. What opportunities for training and professional development has the project provided?This project provided the opportunity for training of a graduate student who complete requirements for a Ph.D. in Rangeland Ecology and Watershed Management at the University of Wyoming in 2016 and is currently writing three or more manuscripts for publication based on project results. This project has also assisted in training two undergraduate student interns with interests in field biology who have assisted with project setup and data collection. How have the results been disseminated to communities of interest?Results have been presented at two scientific conferences that often attract restoration practitioners and agency personnel as well as university researchers. Results have also been disseminated in Extension workshops and as short, informative UW Extension bulletins. We are currently preparing manuscripts for review in scientific journals. What do you plan to do during the next reporting period to accomplish the goals?We are currently completing data analysis and preparing manuscripts for peer review in scientific journals. We may pursue additional funding to continue this project in the future.
Impacts What was accomplished under these goals?
This study tested alternative methods of delineation of seed transfer zones in ecological restoration in order to improve guidelines for revegetation and increase the likelihood of restoration success. Seed transfer zones are regions within which populations may be transferred while minimizing negative effects to population fitness (Hufford and Mazer 2003). Seed transfer zones are used to select seed sources that are adapted to a particular restoration site (Johnson et al. 2004). Accomplishments include: Seed and leaf tissue collections across the range of true mountain mahogany (Cercocarpus montanus Raf.: Rosaceae) were accomplished in the summer and fall of 2013 in Arizona, Utah, New Mexico, Colorado, Wyoming and South Dakota. Seeds were collected from 25 populations for use in common garden study and seed stratification experiments. Leaf tissue was collected from populations to sequence and analyze genetic markers. Additionally, seedlings were grown in the greenhouse and planted in the spring of 2015 to augment the number of living plants in all four common gardens. Common Gardens Common gardens are used to measure genetic variation among populations of a species. Seeds are collected from across a region of interest and grown from seed or as seedlings in a common environment. Common gardens isolate genetic differences by removing environmental variation, and test if populations are adapted to local environmental conditions. We established four common gardens installed along the Front Range Region. Planting multiple common gardens in a variety of habitat types (known as reciprocal transplants), explicitly tests local adaptation, defined as the higher fitness of local populations than non-local populations in two or more study sites. Common gardens were installed using a complete randomized block design in the fall of 2013. Greenhouse grown seedlings were planted in the spring of 2015 to augment the data set. Germination, growth rate, biomass, and survival data were measured in 2014 and 2015 and completed in spring and summer of 2016. Seed Stratification Populations of mountain mahogany were also used in a cold stratification experiment. Cold stratification is the process of subjecting seeds to moist and cold conditions, replicating over-wintering conditions. Mountain mahogany occurs from Mexico to Wyoming and is found from approximately 1,200 to 3,000 meters elevation, and has been found to have genetic variation for seed germination requirements in a previous study (Rosner et al. 2003). We created 4 treatments of cold stratification duration: 2, 4, 6, and 10 weeks, including a control of 0 weeks. At the end of each treatment, seeds were removed from cold stratification and were tested for germination for 5 weeks in a growth chamber, and any ungerminated seeds underwent a tetrazolium test from viability. We found significant differences in germination requirements among populations based on latitude and elevation. Genetic Markers Leaf tissue was collected from the 25 populations used in the common garden study, along with 23 additional populations of C. montanus and two populations of curlleaf mountain mahogany, Cercocarpus ledifolius Nutt. Curlleaf mountain mahogany was included to determine if the two taxa hybridized. DNA was extracted using a modified CTAB protocol (Porebski et al. 1997). Samples were digested using restriction enzymes and ligated to DNA barcodes following the amplified restriction fragments for genomic enrichments protocol (Parchman et al. 2012). Individually barcoded samples were sequenced via Illumina next-generation sequencing and over 6000 SNPs were identified with a combination of custom perl scripts, bcftools and samtools. The genomic dataset was analyzed and SNPs determined that genetic differentation varied clinally in this species and that populations separated by geographic barriers such as the Rocky Mountains, represent significantly different genetic variation. Recommendations are that seed transfer should occur within predicted seed zones and to avoid translocation of materials over long distances and large barriers. Literature Cited Hufford, K. M., and S. J. Mazer. 2003. Plant ecotypes: Genetic differentiation in the age of ecological restoration. Trends Ecol. Evol. 18:147-155. Johnson, G., F. C. Sorensen, J. B. St Clair, and R. C. Cronn. 2004. Pacific Northwest Forest tree seed zones: a template for native plants? Nativ. Plants J. 5:131-140. Parchman, T. L., Z. Gompert, J. Mudge, F. D. Schilkey, C. W. Benkman, and C. A. Buerkle. 2012. Genome-wide association genetics of an adaptive trait in lodgepole pine. Mol. Ecol. 21:2991-3005. Porebski, S., L. G. Bailey, and B. R. Baum. 1997. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol. Biol. Report. 15:8-15. Rosner, L. S., J. T. Harrington, D. R. Dreesen, and L. Murray. 2003. Overcoming dormancy in New Mexico mountain mahogany seed collections. J. Range Manag. 56:198-202.
Publications
- Type:
Journal Articles
Status:
Under Review
Year Published:
2017
Citation:
Crow, T.M., S. E. Albeke, C. Alex Buerkle, and K. M. Hufford. Alternative methods to guide species-specific seed transfer in ecological restoration.
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Progress 10/01/14 to 09/30/15
Outputs Target Audience:During 2015, the target audience reached by this project included US Forest Service Personnel at the Manitou Springs Experimental Forest, colleagues at the New Mexico State University John T. Harrington Forestry Research Center, and staff at Mountain Cement in Laramie and Boulder County Parks and Open Space near Boulder, Colorado. These were formal experiential, outreach and extension opportunities including collaborative efforts between personnel at the University of Wyoming, New Mexico State University and Manitou Springs Experimental Forest. We also spoke with attendees at the 2015 joint conference of the High Altitude Revegetation Committee and Central Rockies Chapter of the Society forEcological Restoration in Fort Collins, Colorado. Informal discussions through Extension included landowners, practitioners and industry professionals in the field of ecological restoration. Changes/Problems:We initially had some difficulty establishing common gardens due to high mortality of transplanted seedlings. In 2015, we grew additional seedlings and augmented common gardens to surmount that problem. In addition, the start of this project was originally delayed due to later than expected arrival of the graduate student who now works on this project. He is currently making excellent progress and has passed all preliminary exam requirements for the Ph.D. degree. What opportunities for training and professional development has the project provided?This project has provided the opportunity for training of a graduate student who is completing requirements for a Ph.D. in Rangeland Ecology and Watershed Management at the University of Wyoming. This project has also assisted in training two undergraduate student interns with interests in field biology who have assisted with project setup and data collection. How have the results been disseminated to communities of interest?Results have been presented at two scientific conferences that often attract restoration practitioners and agency personnel as well as university researchers. Results have also been disseminated in short, informative UW Extension bulletins. We are currently preparing manuscripts for review in scientific journals. What do you plan to do during the next reporting period to accomplish the goals?In the spring and fall of 2016, we will continue to analyze genetic and climate datasets and we will also record data from our common gardens to determine whether populations of mountain mahogany are adapted to local environmental conditions. These data will be used to delineate putative and experimental seed transfer zones for true mountain mahogany. We will compare our results from genetic sequence and common garden data to compare the effectiveness of traditional and novel techniques for delineating seed transfer zones, and help conserve important genetic resources of the species.
Impacts What was accomplished under these goals?
This study will test alternative methods of delineation of seed transfer zones in ecological restoration in order to improve guidelines for revegetation and increase the likelihood of restoration success. Seed transfer zones are regions within which populations may be transferred while minimizing negative effects to population fitness (Hufford and Mazer 2003). Seed transfer zones are used to select seed sources that are adapted to a particular restoration site (Johnson et al. 2004). Accomplishments include: Seed and leaf tissue collections across the range of true mountain mahogany (Cercocarpus montanus Raf.: Rosaceae) were accomplished in the summer and fall of 2013 in Arizona, Utah, New Mexico, Colorado, Wyoming and South Dakota. Seeds were collected from 25 populations for use in common garden study and seed stratification experiments. Leaf tissue was collected from populations to sequence and analyze genetic markers. Additionally, seedlings were grown in the greenhouse and planted in the spring of 2015 to augment the number of living plants in all four common gardens. Common Gardens Common gardens are used to measure genetic variation among populations of a species. Seeds are collected from across a region of interest and grown from seed or as seedlings in a common environment. Common gardens isolate genetic differences by removing environmental variation, and test if populations are adapted to local environmental conditions. We have four common gardens installed along the Front Range Region. Planting multiple common gardens in a variety of habitat types (known as reciprocal transplants), explicitly tests local adaptation, defined as the higher fitness of local populations than non-local populations in two or more study sites. Common gardens were installed using a complete randomized block design in the fall of 2013. Greenhouse grown seedlings were planted in the spring of 2015 to augment the data set. Germination, growth rate, biomass, and survival data were measured in 2014 and 2015 and will continue for the 2016 growing season. Seed Stratification Populations of mountain mahogany were also used in a cold stratification experiment. Cold stratification is the process of subjecting seeds to moist and cold conditions, replicating over-wintering conditions. Mountain mahogany occurs from Mexico to Wyoming and is found from approximately 1,200 to 3,000 meters elevation, and has been found to have genetic variation for seed germination requirements in a previous study (Rosner et al. 2003). We created 4 treatments of cold stratification duration: 2, 4, 6, and 10 weeks, including a control of 0 weeks. At the end of each treatment, seeds were removed from cold stratification and were tested for germination for 5 weeks in a growth chamber, and any ungerminated seeds underwent a tetrazolium test from viability. We found significant differences in germination requirements among populations. Genetic markers Leaf tissue was collected from the 25 populations used in the common garden study (Fig. 1), along with 23 additional populations of C. montanus and two populations of curlleaf mountain mahogany, Cercocarpus ledifolius Nutt. Curlleaf mountain mahogany was included to determine if the two taxa hybridized. DNA was extracted using a modified CTAB protocol (Porebski et al. 1997). Samples were digested using restriction enzymes and ligated to DNA barcodes following the amplified restriction fragments for genomic enrichments protocol (Parchman et al. 2012). Individually barcoded samples were sequenced via Illumina next-generation sequencing and over 6000 SNPs were identified with a combination of custom perl scripts, bcftools and samtools. We are analyzing the SNPs to determine the amount of genetic differentiation among populations, and determine the environmental variables that are driving the intraspecific variation. Literature Cited: Hufford, K. M., and S. J. Mazer. 2003. Plant ecotypes: Genetic differentiation in the age of ecological restoration. Trends Ecol. Evol. 18:147-155. Johnson, G., F. C. Sorensen, J. B. St Clair, and R. C. Cronn. 2004. Pacific Northwest Forest Tree Seed Zones: A Template for Native Plants? Nativ. Plants J. 5:131-140. Parchman, T. L., Z. Gompert, J. Mudge, F. D. Schilkey, C. W. Benkman, and C. A. Buerkle. 2012. Genome-wide association genetics of an adaptive trait in lodgepole pine. Mol. Ecol. 21:2991-3005. Porebski, S., L. G. Bailey, and B. R. Baum. 1997. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Mol. Biol. Report. 15:8-15. Rosner, L. S., J. T. Harrington, D. R. Dreesen, and L. Murray. 2003. Overcoming dormancy in New Mexico mountain mahogany seed collections. J. Range Manag. 56:198-202.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Crow, T.M., K. M. Hufford, C. A. Buerkle. 2015. Alternative methods for delineating seed transfer zones: comparisons of genetic and common garden data. National Native Seed Conference. April 13-16. Symposium Genetics of Plant Restoration and Reforestation in a Changing World.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2015
Citation:
Crow, T.M., S. E. Albeke, C. A. Buerkle, and K. M. Hufford. Alternative methods for delineating seed transfer zones for ecological restoration of Cercocarpus montanus: zonal and continuous predictors of seed transfer success. (submitted to J. of Biogeography)
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Progress 10/01/13 to 09/30/14
Outputs Target Audience: During 2014, the target audience reached by this project included US Forest Service Personnel at the Manitou Springs Experimental Forest, colleagues at the New Mexico State University John T. Harrington Forestry Research Center, and staff at Mountain Cement in Laramie and Boulder County Parks and Open Space near Boulder, Colorado. These were formal experiential, outreach and extension opportunities including collaborative efforts between personnel at the University of Wyoming, New Mexico State University and Manitou Springs Experimental Forest. We also spoke with attendees at the annual Botanical Society of America Conference in Boise, Idaho. Informal discussions through Extension included landowners, practitioners and industry professionals in the field of ecological restoration. Changes/Problems: This project was delayed by the late admission of a graduate student working on this project. In 2013, we successfully recruited a graduate student who has made significant progress to meet project objectives and who will, through completion of this research, earn his Ph.D. in Rangeland Ecology and Watershed Management at the University of Wyoming. What opportunities for training and professional development has the project provided? Training and opportunites for professional development include graduate student Taylor Crow and undergraduate intern Cody Starosta. One-on-one mentoring activities are routine to discuss experimental design, laboratory protocols, and project management. Taylor Crow presented initial results at the national conference of the Botanical Society of America. Cody Starosta worked with Taylor Crow to establish experimental sites and monitor common gardens. How have the results been disseminated to communities of interest? Results have been disseminated via annual reports sent to collaborators, presentations at national conferences, and informal discussions with US Forest Service personnel as well as participants in regional and local workshops via University of Wyoming Extension. What do you plan to do during the next reporting period to accomplish the goals? In spring 2015, we will augment our common gardens with seedlings to increase the total number of plants in the experiment and track fitness of seedlings from 25 populations in a common environment . Planting seedlings will bypass the germination bottleneck, and increase the sample size for phenotypic measures. Data collections at each common garden will continue through the 2015 and 2016 growing season to measure genetic differentiation for quantitative traits. We will use the data generated from common gardens and molecular markers to delineate seed transfer zones for true mountain mahogany. True mountain mahogany is a keystone species and mountain mahogany shrublands represent critical wildlife habitat in regions under pressure from human development. Seed transfer zones will assist restoration and management of these shrublands. Seed zones based on field studies will be compared with results for molecular markers as well as species distribution models to determine if alternative methods for seed zone delineation can accuarately describe adaptive genetic variation and predict suitable regions for transfer of seeds in ecological restoration. Results will improve restoration practice for this species and have potential application for a suite of species targeted in restorationif alternative methods for delineation are suitable to guide seed transfer in revegetation.
Impacts What was accomplished under these goals?
In 2013-14, we completed collections of seed and leaf tissue from across the range of true mountain mahogany (Cercocarpus montanus Raf.: Rosaceae) representing populations from Arizona, Utah, New Mexico, Colorado, Wyoming and South Dakota. Seeds representing 25 populations were planted in a series of four common gardens and also tested in seed stratification experiments to determine if cold chill requirements differ across the species' range. Monitoring of common gardens is ongoing and will include new plantings in 2015. Leaf tissue was collected from the 25 populations represented in the common garden study, along with 23 additional populations of C. montanus and two populations of curlleaf mountain mahogany (Cercocarpus ledifolius Nutt). Curlleaf mountain mahogany was included in genetic analyses to determine if the focal populations of C. montanus hybridize with C. ledifolius. DNA was extracted from 40 individuals per population using a modified CTAB protocol. Samples were subsequently digested using restriction enzymes and ligated to DNA barcodes following a protocol for genomic enrichment using amplified restriction fragments. Individually barcoded samples were sequenced via Illumina next-generation sequencing and over 6000 SNPs were identified with a combination of custom perl scripts, bcftools and samtools. Genetic markers will be analyzed to characterize population genetic structure and diversity across the species north-south range. Results will be compared with field studies of common gardens to determine if population genetic structure reflects adaptive differentiation, testing the utility of genetic markers for delineation of seed transfer zones at a range of geographic scales.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2014
Citation:
Crow, T.M. and K.M. Hufford. 2014. Improving marginal habitat restoration in Western rangelands: ecological genetic and landscape approaches to mountain mahogany shrubland reclamation. Botany 2014 New Frontiers in Botany, Boise, ID, July 2014 Botanical Society of America conference.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2015
Citation:
Crow, T.M., K.M. Hufford, and C.A. Buerkle. 2015. Alternative methods for delineating seed transfer zones: comparisons of genetic and common garden data. National Native Seed Conference, April 13-16, Santa Fe, New Mexico.
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Progress 01/01/13 to 09/30/13
Outputs Target Audience: Target Audience: During 2013, the target audience reached by this proposal included US Forest Service Personnel at the Manitou Springs Experimental Forest, colleagues at the New Mexico State University John T. Harrington Forestry Research Center, and staff at Mountain Cement in Laramie and Boulder County Parks and Open Space near Boulder, Colorado. These were informal experiential, outreach and extension opportunities that occurred at the initiation of the field component of this research study. Changes/Problems: This project was delayed by the late admission of a graduate student working on this project. In 2013, we successfully recruited a graduate student who has begun work to meet project objectives and who will, through completion of this research, earn his PhD in Rangeland Ecology and Watershed Management at the University of Wyoming. What opportunities for training and professional development has the project provided? Opportunities for training and professional development include one-on-one mentoring with a PhD student as well as individual study by that student to learn GIS methods for analysis of climate and physical site variation within the range of the field study. Additional mentoring occurred with two undergraduate research interns who have assisted with setup of field components of this research. How have the results been disseminated to communities of interest? Initial outreach efforts have begun at each experimental site to discuss seed transfer zones with individuals who are working in private industry (Laramie, Mountain Cement Company), and city open area land management (Boulder County Parks & Open Space) and to initiate collaborative research with US Forest Service Personnel and colleagues at New Mexico State University. Outreatch activities include informal discussions, field meetings and participation in field research. What do you plan to do during the next reporting period to accomplish the goals? In the next reporting period we plan to gather data for Cercocarpus montanus germination, survival and growth from the field and greenhouse studies . We will continue to acquire genetic data and, once sufficient data is available, we will begin analyses to develop estimates of marker-based seed zones for this species. In addition, work will be accomplished to develop ArcGIS-based estimates of climate and soil variation for comparison of environmental factors to genetic and field data to determine the efficacy of seed zones (and different types of zones) for restoration of this species.
Impacts What was accomplished under these goals?
1. Investigate genetic diversity of Cercocarpus montanus and delineate seed zones based on molecular marker differentiation. Leaf tissue from 30 or more individuals and a total of 46 populations of Cercocarpus montanus were collected for genetic analyses along a north-south transect from northern Wyoming to New Mexico. Data acquisition is underway for 25 of these collections to meet objective 1 and characterize genetic diversity for molecular delineation of seed zones in this species. 2. Determine the relationship of marker-based transfer zones and successful establishment under different climate and habitat conditions. We have established reciprocal transplant plots at in Mora, New Mexico (John T Harrington forestry research center); Manitou Springs and Lyons, Colorado (Manitou Experimental Forest and Boulder County Parks & Open Space); and Laramie, Wyoming to test for evidence of local adaptation among different seed sources of Cercocarpus montanus. A minimum of 1,500 seeds are planted at each location and results from this field experiment will be compared to estimates of genetic divergence (Objective 1) to determine if marker-based transfer zones predict successful establishment of plants as seen in the field study. In addition, greenhouse transplants of seed sources between soil types are also underway to meet the same objective of understanding environmental factors that drive successful establishment of Cercocarpus montanus in a controlled environment. 3. Determine the efficacy of matching transfer zones to the local environment for restoration outcomes. Efficacy of transfer zones will be determined through comparisons of data derived from activities that meet objectives 1 and 2. Marker-derived transfer zones will also be compared to estimates of climate- and soil-based transfer zones. To this end, climate- and soil-based estimates of transfer zones are underway using ArcGIS software to examine the region in which field and genetic studies have been established.
Publications
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Progress 01/01/12 to 12/31/12
Outputs OUTPUTS: Outputs to date include collections of Cercocarpus montanus (C. montanus) in three states, seed germination studies of C. montanus, initial DNA work of C. montanus collections, mentoring a Conservation and Land Management Intern, and May 15 - ongoing mentoring three undergraduates as part of C. montanus collections and seed germination research Event: Wyoming Native Seeds Workshop, Presentation April 3, 2012. PARTICIPANTS: Individuals: Georgia Thomas, CLM Intern; Ariana Roe, UW undergraduate student; Rachel Strawn, UW undergraduate student; Amy Jacobs, UW undergraduate student. Partner organization: Bureau of Land Management via the Seeds of Success program. TARGET AUDIENCES: 1. Federal land management agencies that oversee land reclamation and restoration (e.g., BLM). 2. Commercial seed industries that produce native plant seeds for reclamation and restoration. 3. Restoration practitioners who work directly on restoration projects in the field. PROJECT MODIFICATIONS: Project is currently underway but the arrival of a graduate student who will work on the project has been delayed.
Impacts Impacts include new data for C. montanus germination and original DNA work for the species. Participants have learned field collection and laboratory germination techniques and improved their understanding of scientific publications and scientific methodology. As this is a new project, we will report additional outcomes in the future.
Publications
- No publications reported this period
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