Progress 09/01/08 to 08/31/13
Outputs
Target Audience: K-State Research and Extension, KSU Agricultural Research Centers, biofuel industries, cattle ranchers, other faculty members, Manhattan KS USDA Plant Material Center and other USDA Plant Material Centers, other scientists and faculty members. We have taken a number of informal education groups to our reciprocal garden research platform including a visit by a state-wide high school group interested in bluestem ecotypes and disease, a visit by a group from the national meeting of the Plant Pathology Society, use of the plant materials for use in Junction City High School Genetics classes, planned visit by a group from a national agricultural statistics conference, three high school students carrying out independent research using our reciprocal garden sites and plant materials, about 5 undergrads per year conducting research on the garden sites, tours as part of the USDA Plant Material Field Days. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Five graduate students were trained and supported from the grant. An additional 4 students took advantage of the ecotype experimental platform to conduct work as part of their theses. Student Theses 1. Jacob T. Olsen, M.S. in Biology, Fort Hays State University Thesis: Gas exchange and leaf anatomy of Andropogon gerardii ecotypes over a climatic gradient of the Great Plains.2012. 2. Miranda Gray MS thesis KSU, Manhattan, KS. GENOMIC DIFFERENTIATION OF BIG BLUESTEM (ANDROPOGON GERARDII) ALONG THE GREAT PLAINS’ ENVIRONMENTAL GRADIENT 2012. 3. Jennifer Shelton MS thesis KSU, Manhattan, KS.2012. EPICUTICULAR WAX CHEMISTRY, MORPHOLOGY, AND PHYSIOLOGY IN SAND BLUESTEM, ANDROPOGON GERARDII SSP. HALLI, AND BIG BLUESTEM, ANDROPOGON GERARDII SSP. GERARDII 4. Rachel Goad MS thesis SIU, Carbondale Illinois.2012. Patterns in Aboveground Net Primary Productivity in an Assembling[LJ1] Tallgrass Prairies Communities Across a Precipitation Gradient 5. Meredith Mendola. MS thesis SIU, Carbondale Illinois. 2013. Ecotypic variation in root biomass and belowground net primary productivity of Andropogon gerardii 6. Donatelli Juliette MS thesis SIU, Carbondale Illinois. Forage quality of Andropogon gerardii across a precipitation gradient.2013. 7. Laurel Wilson, MS thesis SIU, Plant community response to regional sources of dominant grasses in grasslands restored across a longitudinal gradient. 2013. 8. Jing Gan MS thesis KSU, Manhattan, KS. 9. Ke Zhang-in progress Other training: Graduate student grants and awards: Miranda Gray: Bringing Genomics Outdoors: A Look into the Drought Transcriptome and Differential Physiological Response of Dominant Prairie Grass Phenotypic Variants from Opposing Precipitation Regimes. Kansas Native Plant Society Award to Student Miranda Gray $1000: Jacob Olsen: Gas exchange differnces among bluestem ecotypes across the Great Plains precipitation gradient. Kansas Native Plant Society Award to Student Jacob Olsen $1000: Miranda Gray-ASPB best poster, 2012. Bringing Genomics Outdoors: A Look into the Drought Transcriptome and Differential Physiological Response of Dominant Prairie Grass Phenotypic Variants from Opposing Precipitation Regimes Undergraduate grants and awards Keri L. Caudle, Fort Hays State University Biology. Outstanding Undergraduate Student, FHSU College of Health and Life Sciences, Botanical Society of America “Spotlight Member,” March to May of 2012 Matthew Gaillart-KSU, Biology. Recipient of NSF URM fellowship, won best poster at KSU Undergraduate Research Forum, 2nd best poster at REU poster conference. Matthew Gaillart. Research Grant: Outcomes of competition among ecotypes. Kansas Academy of Science research grant $1,000 Matthew Gaillart. Research Grant for bluestem sequencing from the KUMC seed grant program ($6,000 value) Training Opportunities Several students took part in courses at the Integrative Genomics facility including courses on RT PCR, Microarrays, RNA seq, and R. [LJ1]Other places you refer to it as reassembling How have the results been disseminated to communities of interest? We have taken a number of informal education groups to our reciprocal garden research platform including a visit by a state-wide high school group interested in bluestem ecotypes and disease, a visit by a group from the national meeting of the Plant Pathology Society, use of the plant materials for use in Junction City High School Genetics classes, planned visit by a group from a national agricultural statistics conference, three high school students carrying out independent research using our reciprocal garden sites and plant materials, about 5 undergrads per year conducting research on the garden sites, tours as part of the USDA Plant Material Field Days. Collectively, we have made tens of presentations at regional, national, and international meetings. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The central grassland of North America is one of the most productive rangelands in agricultural productivity in KS alone. Changes in amount of precipitation will likely impose critical environmental stress and may compromise future grassland productivity and sustainability. We aim to investigate ecotypic variation in big bluestem Andropogon gerardii under natural and experimentally reduced precipitation across prairie from KS to IL, and to identify the genetic bases of underlying adaptive variation in drought tolerance, which will ultimately be used to predict the functional response to climate change. We will determine adaptive drought response using single and mixed ecotype assemblages of A. gerardii grown in ambient home environments, reciprocally planted across the precipitation gradient, and in response to reduced precipitation. Specifically, we characterized: (i) phenotypic response of ecotypes in their drought-related physiological processes (ii) genetic diversity among ecotypes and widely used cultivars in populations from KS to IL, (iii) functional genomic consequences of ecotypic differentiation at the level of gene expression; and (iv) ecosystem consequences of ecotypic variation in drought tolerance. In this experimental design, we have grown in 4 climate locations plants of 3 ecotypes (each comprised of 4 separate populations) in replicated blocks (n=10) with each plant growing singly in plastic and in artificially assembled seeded communities (n=4 per planting location, 4x4 m plots). Rainout shelters were constructed and put in place spring 2011. We made the following functional, morphological and performance responses of the bluestem ecotypes in all four planting locations in the seeded and single spaced plantings and reciprocal gardens in multiple years: leaf-level gas exchange and photosynthesis, ecotypic-specific plant cover to assess differences in establishment and growth, end of season biomass, phenological and seed production differences, and collected leaf samples from all sites for genotyping and transcriptome sequencing (454 and Illumina). We measured soil moisture and precipitation throughout the season. We developed AFLP genotyping markers from 24 plants per population, 12 populations comprised of 3 ecotypes. We plan to analyze bluestem seed from additional populations in order to place our focal populations into a broader genetic and ecological context to test for genetic structure of big bluestem across the Great Plains. Beginning to expand our marker platform using GBS with Dr Jesse Poland. We began collaborations with Dr Sue Brown (KSU Bioinformatics Center) in the development of a de novo reference transcriptome hybrid assembled from 454 and Illumina reads. The hybrid-assembled transcriptome will be a critical resource for future mapping of Illumina reads from drought and control samples. In addition, we held coordination and planning meetings annually involving PIs and student presentations. We met all objectives as proposed. Big bluestem Andropogon gerardii is a widely-distributed dominant C4 grass, whose productivity is dependent upon precipitation. With wide distribution across a sharp precipitation gradient (400-1200mm yr-1 in Kansas to Illinois), we expect ecotypic variation in drought tolerance and potentially, local drought adaptation. A better understanding of ecotypic variation will help predict how a dominant forage grass may respond to climate change and will inform prairie restoration. We investigated the linkage of phenotypic and genetic variation by using reciprocal common gardens established across the precipitation gradient of the Great Plains. Sites were planted in Carbondale, Illinois; Manhattan and Hays, Kansas; and a site in Colby, KS at the species’ range margin (to test ecotype tolerance into drier areas). We measured phenotypic variation in drought tolerance across ecotypes and sites. As genetic variation may be critical for predicting a species’ ability to adjust/adapt to climate change, we assessed genetic diversity and population structure and population differentiation using genome scans of Andropogon gerardii in the 12 source prairies populations also used in the reciprocal garden experiment. We complemented the marker studies with transcriptome profiling to determine to what extent ecotypes can adjust and cope with abiotic stresses. Our results demonstrate a strong phenotypic cline in drought tolerance among the three regional ecotypes. The xeric ecotype (central KS) exhibits local adaptation to drought based on the reciprocal garden results. Establishment and cover in seeded plots showed a significant ecotype (p<0.0001), site (p<0.0001) and interaction effect (p<0.0001). The central KS ecotype had disproportionately greater cover in western regions relative to the Illinois and eastern KS ecotypes (GXE), indicating local adaptation to drought. Several lines of evidence support genetic differentiation and genetic structuring of regional ecotypes. Eight ecotype-specific loci were identified to be under divergent selection using BAYESCAN, and each of these AFLP marker loci frequencies were shown to shift depending on precipitation. In spite of the genetic differentiation found among regional ecotypes, AMOVA analyses partition the greatest genetic variation within prairies (80%), suggesting sufficient genetic variation for adaptive potential. To assess transcriptional response for the xeric and mesic ecotypes under severe abiotic stress growing in western KS, we further used 454 sequencing and de novo assembly of transcriptomes from the xeric central KS and mesic Illinois ecotype, complemented with Illumina RNA-sequencing. A total of 245 transcripts were differentially expressed (~1.8% of total transcripts) between Central KS and Illinois ecotypes under two-week natural drought. Notably, the mesic Illinois ecotype appears to mount a weaker transcriptional response, primarily consisting of genes involved in abiotic stress as an inducible, short-term response to water stress. In contrast, the xeric central KS ecotype shows a stronger, more varied transcriptional response that matched its superior growth response. These phenotypic and genetic variation studies provide new insights into how this dominant forage grass may withstand environmental change and may inform prairie restoration strategies in the Great Plains. Reciprocal Garden Research Platform. The development of this resource has been invaluable for multiple researchers and has provided spin-offs in multiple new directions with new collaborators using the platform. We are developing a new Genotyping by Sequencing approach to produces thousands of molecular markers for big bluestem in collaboration with Jesse Poland. We have developed de novo transcriptome resources for the two most divergent ecotypes (xeric and mesic). This resource is a valuable tool to identify transcriptional differences and SNPs associated with drought tolerance traits. We used 454 sequencing and Newbler de novo transcriptome assembly, complemented with additional Illumina RNA-sequencing. Studies on phenotypic and genetic variation will guide the development of germplasm that is locally adapted to drought. We received three seed grants to defray costs of sequencing. Lipidomics. Through several pilot studies using GC-MS, we are detecting very distinct lipid profiles among bluestem ecotypes. These results indicate that big bluestem ecotypes are quite distinctive in both epicuticular wax load and composition. This may be important for drought tolerance. Biofuel Potential-We began collaborations with perennial grass biofuel engineers Wang and Yuan to determine the biofuel potential of big bluestem. With plant material from the reciprocal gardens, we have provided the first comprehensive studies of bluestem biofuel potential and the control of environment and ecotype in controlling yield and suitability for biofuels.
Publications
- Type:
Other
Status:
Accepted
Year Published:
2012
Citation:
Gan Jing, Wenqiao Yuan, Loretta Johnson, Donghai Wang a, Richard Nelson, Ke Zhang. 2012
Hydrothermal conversion of big bluestem for bio-oil production: The effect of ecotype and planting location. Bioresource Technology 116 (2012) 413420
Zhang, Ke Loretta Johnson, Richard Nelson, Wenqiao Yuan, Zhijian. Pei, and Donghai Wang. 2012. Chemical and elemental composition of big bluestem as affected by ecotype and planting location along the precipitation gradient of the Great Plains. Industrial Crops and Products 40: 210218
- Type:
Theses/Dissertations
Status:
Accepted
Year Published:
2012
Citation:
1. Jacob T. Olsen, M.S. in Biology, Fort Hays State University
Thesis: Gas exchange and leaf anatomy of Andropogon gerardii ecotypes over a climatic gradient of the Great Plains.2012.
2. Miranda Gray MS thesis KSU, Manhattan, KS. GENOMIC DIFFERENTIATION OF BIG BLUESTEM (ANDROPOGON GERARDII) ALONG THE GREAT PLAINS ENVIRONMENTAL GRADIENT 2012.
3. Jennifer Shelton MS thesis KSU, Manhattan, KS.2012.
EPICUTICULAR WAX CHEMISTRY, MORPHOLOGY, AND PHYSIOLOGY IN SAND BLUESTEM, ANDROPOGON GERARDII SSP. HALLI, AND BIG BLUESTEM, ANDROPOGON GERARDII SSP. GERARDII
4. Rachel Goad MS thesis SIU, Carbondale Illinois.2012.
Patterns in Aboveground Net Primary Productivity in an Assembling Tallgrass Prairies Communities Across a Precipitation Gradient
- Type:
Theses/Dissertations
Status:
Accepted
Year Published:
2013
Citation:
5. Meredith Mendola. MS thesis SIU, Carbondale Illinois. 2013.
Ecotypic variation in root biomass and belowground net primary productivity of Andropogon gerardii
6. Donatelli Juliette MS thesis SIU, Carbondale Illinois.
Forage quality of Andropogon gerardii across a precipitation gradient.2013.
7. Laurel Wilson, MS thesis SIU, Plant community response to regional sources of dominant grasses in grasslands restored across a longitudinal gradient. 2013.
8. Jing Gan MS thesis KSU, Manhattan, KS. HYDROTHERMAL CONVERSION OF LIGNOCELLULOSIC BIOMASS TO BIO-OILS
9. Ke Zhang-in progress The effects of ecotype and planting location on properties and bio-fuel yield of big bluestem
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
Baer, S.G., B.R. Maricle, N. Bello, K.L. Caudle, M. Galliart, D.J. Gibson, M. Gray, R.K. Goad, E. McCrea, M.L. Mendola, J.T. Olsen, S. Sabates, H. Tetrault, L. Wilson, and L.C. Johnson. Discerning the presence and ecological consequence of ecotypic variation: A dilemma for restoration. Invited presentation at the symposium, Changing prairies: science and practice in grassland restoration and reconstruction, at the 5th World Conference of Ecological Restoration, Madison, WI, October 2013
Switchgrass Genomics Conference. Johnson, L, S.G. Baer, B.R. Maricle, M. Gray , H. Tetreault , N. Bello, S. Brown, T. Morgan, J. Shelton , and S. Chellapilla. Genetic differentiation, transcriptome variation, and local adaptation of an ecologically dominant prairie grass Andropogon gerardii (big bluestem) occurring along the climate gradient in Midwest U.S. grasslands. Madison, WI, Sept 9-10.
Johnson, L, M. Gray , H. Tetreault , K. Garrett, E. Akhunov, P. St. Amand, N. Bello, J. Bryant*, N. An , B.R. Maricle, J. Olsen , S.G. Baer, R.K. Goad , A. De La Cruz , T. Morgan, and M. Knapp. Ecotypic variation in drought tolerance and genetic diversity of the ecologically dominant grass big bluestem (Andropogon gerardii) across the Great Plains precipitation gradient: Implications for climate change and restoration. Presented at the Americas Grasslands national meeting, Manhattan, KS, August 2013
Dr. Loretta Johnson, Biology Dr. Wengaio Yuan, Biol. and Ag. Engineering Dr. Donghai Wang, Biol. and Ag. Engineering Dr. Richard Nelson, Center for Sustainable Energy. Big bluestem as a potential bioenergy grass. Energy Sustainability Conference. KSU April 2013
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2011
Citation:
Olsen, J.T. , H.M. Tetrault , R.K. Goad , M.L. Mendola*, L.C. Johnson, S.G. Baer, and B.R. Maricle. Photosynthetic variation of big bluestem and sand bluestem influenced by ecotype and precipitation. Presented at the Botanical Society of America Annual Meeting, St. Louis, MO, July 2011 (oral)
Olsen, J.T. , H.M. Tetreault , R.K. Goad , M.L. Mendola*, L.C. Johnson, S.G. Baer, and B.R. Maricle. Variation of photosynthesis in big bluestem and sand bluestem reciprocally planted across a precipitation gradient. Presented at the Kansas Academy of Science Annual Meeting, Baldwin City, KS, April 2011 (oral)
Sendor, G. *, J. Donatelli , D.J. Gibson, S.G. Baer, & L. Johnson. 2011. Population variation in relative fitness of Andropogon gerardii (Big bluestem). SIUC Undergraduate Research Forum, April 11, 2011. (poster presentation)
Olsen, J.T. , H.M. Tetrault , R.K. Goad , M.L. Mendola*, L.C. Johnson, S.G. Baer, and B.R. Maricle. Photosynthetic variation of big bluestem and sand bluestem influenced by ecotype and precipitation. Presented at the Botanical Society of America Annual Meeting, St. Louis, MO, July 2011 (oral presentation)
Olsen, J.T. , H.M. Tetreault , R.K. Goad , M.L. Mendola*, L.C. Johnson, S.G. Baer, and B.R. Maricle. Variation of photosynthesis in big bluestem and sand bluestem reciprocally planted across a precipitation gradient. Presented at the Kansas Academy of Science Annual Meeting, Baldwin City, KS, April 2011 (oral presentation)
KSU Graduate School SUROP (Summer Undergraduate Research Opportunities) seminar, July, 2011. Angel DeLaCruz*, Phenotypic variation and Chorophyll Absorbance in Big Bluestem (Andropogon gerardii) ecotypes across the Great Plains precipitation gradient (oral presentation)
Goad R.K. , S.G. Baer, L.C. Johnson, and B.R. Maricle. Patterns in aboveground net primary productivity in prairie reciprocally restored with dominant grasses from across a precipitation gradient. Presented at the Grasslands in a Global Context meeting, Manhattan, KS, September 2011 (poster presentation)
Tetreault, H , C. Rodewald*, S. Baer, B. Maricle, N. Bello, T. Morgan, R. Goad , J. Olsen , and L. Johnson. Local drought adaptation of the ecologically dominant prairie grass big bluestem Andropogon gerardii: Contribution of genotype and environment to phenotypic variation. Ecological Society of America Annual Meeting, Austin, TX, August 2011 (poster presentation)
- Type:
Other
Status:
Accepted
Year Published:
2013
Citation:
Sabates, S., M. Galliart, S. Baer, N. Bello, K. Caudle, B. Maricle, E. McCrea, D. Gibson, L. Wilson, and L. Johnson. 2013. How does phenology and seed production of big bluestem (Andropogon gerardii) vary across the climate gradient of the Great Plains? A reciprocal garden approach to distinguish roles of site and ecotype. Botanical Society of America Annual Meeting, July 27-August 1, New Orleans, Louisiana. Evolution Meeting, Snowbird, UT, June ,2013 (poster)
Maricle, B. R., M. M. Gray, J. Bryant, A. Jensen, A. de la Cruz, K. L. Caudle, J. T. Olsen, S. G. Baer, M. Knapp, and L. C. Johnson. 2013. A possible mechanism for increased performance of a xeric adapted big bluestem (Andropogon gerardii) ecotype: Nitrogen and chlorophyll content of leaves in reciprocal gardens across the Great Plains. Botanical Society of America Annual Meeting, July 27-August 1, New Orleans, Louisiana and Americas Grasslands Conference, August 13-14. Manhattan, Kansas. (poster)
Caudle, K. L., L. C. Johnson, S. G. Baer, and B. R. Maricle. 2013. Influence of precipitation on trichome densities in big bluestem (Andropogon gerardii) ecotypes in Great Plains reciprocal gardens. Botanical Society of America Annual Meeting, July 27-August 1, New Orleans, Louisiana and Americas Grasslands Conference, August 13-14. Manhattan, Kansas.
Johnson, L, M. Gray , H. Tetreault , K. Garrett, E. Akhunov, P. St. Amand, N. Bello, J. Bryant*, N. An , B.R. Maricle, J. Olsen , S.G. Baer, R.K. Goad , A. De La Cruz , T. Morgan, and M. Knapp. Ecotypic variation in drought tolerance and genetic diversity of the ecologically dominant grass big bluestem (Andropogon gerardii) across the Great Plains precipitation gradient: Implications for climate change and restoration. Presented at the Americas Grasslands meeting, Manhattan, KS, August 2013. (poster)
- Type:
Other
Status:
Accepted
Year Published:
2013
Citation:
Maricle, B.R., K.L. Caudle*, K.J. Lindsey , S.G. Baer, and L.C. Johnson. Effects of extreme drought on photosynthesis and water potential of Andropogon gerardii (big bluestem) ecotypes in common gardens across Kansas. Presented at the Kansas Academy of Science Annual Meeting, Overland Park, KS, April 2013 (oral)
Sabates, S., M. Galliart, S. Baer, N. Bello, K. Caudle, B. Maricle, E. McCrea, D. Gibson, L. Wilson, and L. Johnson. 2013. How does phenology and seed production of big bluestem (Andropogon gerardii) vary across the climate gradient of the Great Plains? A reciprocal garden approach to distinguish roles of site and ecotype.KSU University Wide Research Forum, Best poster award.
Loretta C Johnson, Miranda Gray, Alina Akhunova, Susan J. Brown, Sanjay Chellapilla, Theodore Morgan, Karen Garrett, Hanquan Liang and Eduard Akhunov, Loretta Phenotypic and genetic variation in a dominant forage and biofuel grass along the Great Plains' precipitation gradient: a reciprocal garden approach. 2013 PAG, San Diego.(poster)
Shelton, Jennifer , Miranda M. Gray , Susan J. Brown, Sanjay Chellapilla, Alina R. Akhunova, Eduard D. Akhunov, Hanquan Liang, Loretta C. Johnson De novo transcriptome profiling of two edaphically and phenotypically divergent grasses: dominant forage grass big bluestem Andropogon gerardii and drought-tolerant sand bluestem Andropogon gerardii ssp. Hallii. 2013 PAG, San Diego. (poster)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2012
Citation:
Thilani N. Samarakoon, Jennifer M. Shelton , Richard Jeannotte, Zhihong Song, Nora Bello, Basil Nikolau, Ruth Welti, Loretta Johnson. A Gas Chromatography/Mass Spectrometry Reveals Contrasts in Epicuticular Wax Composition of Big Bluestem (Andropogon gerardii) and edaphic variety Sand Bluestem (A. gerardii var halli.)ASPB, Oral Presentation, Lincoln Nebraska, March 2012.
Shelton, Jennifer M , Ravenek Janneke , Samarakoon Thilani, Jeannotte Richard, Welti Ruth, Nikolau, Basil J, Song Zhihong, and Johnson, Loretta. Divergence in leaf morphology and physiology in Andropogon gerardii var. hallii (sand bluestem) a locally adapted variety of Andropogon gerardii (big bluestem) ASPB, Poster Presentation, Lincoln Nebraska, March 2012.
Olsen, J.T. , H.M. Tetrault , R.K. Goad , M.L. Mendola , L.C. Johnson, S.G. Baer, and B.R. Maricle. Photosynthetic variation of big bluestem and sand bluestem influenced by ecotype and precipitation. Presented at the Botanical Society of America Annual Meeting, Columbus July 2012 (oral presentation)
Johnson, L, M. Gray , H. Tetreault , K. Garrett, E. Akhunov, P. St. Amand, N. Bello, J. Bryant*, N. An , B.R. Maricle, J. Olsen , S.G. Baer, R.K. Goad , A. De La Cruz , T. Morgan, and M. Knapp. Ecotypic variation in drought tolerance and genetic diversity of the ecologically dominant grass big bluestem (Andropogon gerardii) across the Great Plains precipitation gradient. Presented at the Ecological Society of America Annual Meeting, Portland, OR, August 2012. (oral)
- Type:
Other
Status:
Accepted
Year Published:
2012
Citation:
Olsen, J.T. , R.K. Goad , M.L. Mendola*, H.M. Tetrault , L.C. Johnson, S.G. Baer, and B.R. Maricle. Photosynthetic gas exchange of big bluestem (Andropogon gerardii) ecotypes across a precipitation gradient of the Great Plains. Presented at the Botanical Society of America Annual Meeting, Columbus, OH, July 2012. (oral)
Olsen, J.T. , R.K. Goad , M.L. Mendola*, H.M. Tetrault , L.C. Johnson, S.G. Baer, and B.R. Maricle. Gas exchange of Andropogon gerardii (big bluestem) ecotypes across a climatic gradient of the Great Plains. Presented at the Kansas Academy of Science Annual Meeting, Wichita, KS, March 2012 (oral)
Gray, Paul St. Amand, Loretta Johnson, Eduard Akhunov, Karen Garrett, Mary Knapp, Alina Akhunova, Sanjay Chellapilla, Ted Morgan, Guihua Bai. Genetic differentiation of Andropogon gerardii(big bluestem) ecotypes along a natural precipitation gradient Miranda Botanical Society of America. Columbus July 2012 (oral Presentation)
KSU Graduate School SUROP (Summer Undergraduate Research Opportunities) seminar, July, 2012. Jessica Gaffney*, Variations in polyploidy in Big Bluestem (Andropogon gerardii) ecotypes across the Great Plains (oral)
KSU Graduate School REU (Summer Undergraduate Research Opportunities) seminar, July, 2012. Amber Jensen*, Big bluestem ecotypic differences in leaf nitrogen content as a possible mechanism for increased performance of western Kansas ecotypes in reciprocal gardens planted across the Great Plains..(oral)
KSU URM (Undergraduate Research Mentoring Opportunities) Sofia Sabates*. July 2012 Phenology and seed production in Big Bluestem (Andropogon gerardii) ecotypes in Great Plains reciprocal gardens: the role of site and ecotype (poster)
K. Zhang, L. Johnson, R. Nelson, W. Yuan, ZJ. Pei, and D. Wang. 2012. Chemical and elemental composition of big bluestem as affected by ecotype and planting location along the precipitation gradient of the Great Plains. 2012 ASABE Annual Meeting, 6/29 to 7/1, 2012, Dallas, TX. Paper No.121338462
- Type:
Other
Status:
Accepted
Year Published:
2012
Citation:
Ecotypic variation in drought tolerance and genetic diversity of an ecologically dominant grass big bluestem (Andropogon gerardii) across the Great Plains precipitation gradient Gray , Miranda, St Amand Paul, Tetreault, Hannah ; Baer, Sara; Maricle, Brian; Morgan, Ted; Goad, Rachel ; Olsen , Jake and Johnson, Loretta. Ecological Society of America. Portland Oregon, Aug. 2012. (oral)
Meredith L Mendola , Sara G Baer, Loretta C Johnson. Ecotype and climate controls on belowground ecosystem processes. Oral Presentation. Ecological Society of America. Portland Oregon, Aug. 2012.
Gan Jing, Wenqiao Yuan, Loretta Johnson, Donghai Wang a, Richard Nelson, Ke Zhang.
Hydrothermal conversion of big bluestem for bio-oil production: The effect of ecotype and planting location. Presentation, DOT SunGrant Conference, Oct 2012. New Orleans
Zhang, Ke Loretta Johnson, Richard Nelson, Wenqiao Yuan, Zhijian. Pei, and Donghai Wang. Chemical and elemental composition of big bluestem as affected by ecotype and planting location along the precipitation gradient of the Great Plains. Presentation, DOT SunGrant Conference, Oct 2012. New Orleans
Gray , Miranda, St Amand Paul, Tetreault, Hannah ; Baer, Sara; Maricle, Brian; Morgan, Ted; Goad, Rachel ; Olsen , Jake and Johnson, Loretta Phenotypic and genetic variation in a dominant prairie grass along the Great Plains' precipitation gradient: a reciprocal garden approach. Poster, Ecological Genomics Symposium, Oct 26-28, KC.
Gaffney, Jessica*; Sabates, Sofia*; Baer, Sara; Maricle, Brian; McCrea, Evan*; Bello, Nora M. and Johnson, Loretta Variations in polyploidy in Big Bluestem (Andropogon gerardii) ecotypes across the Great Plains. Poster, Ecological Genomics Symposium, Oct 26-28, 2012 KC.
Sabates, Sofia*; Baer, Sara; Caudle, Keri*; Harmoney, Keith; Maricle, Brian; McCrea, Evan*; and Johnson, Loretta. Phenology and seed production in Big Bluestem (Andropogon gerardii) ecotypes in Great Plains reciprocal gardens: the role of site and ecotype Poster, Ecological Genomics Symposium, Oct 26-28,2012. KC.
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Progress 09/01/11 to 08/31/12
Outputs
OUTPUTS: The major activity for 2011-2012 was data analyses, dissemination at national scientific meetings, and manuscript preparation and submissions, and plans for grant submissions for continued funding. We also continued to make measurements on our bluestem ecotype reciprocal gardens. 2012 was of significance as it was the worst drought in the Great Plains in the last 50 years. In Spring 2012, we continued our third year of measurements.The gardens were established in each of the precipitation zones of the Great Plains to measure genotype by environment interaction and to detect evidence of adaptive differentiation across the precipitation gradient at three sites (Carbondale Illinois, Manhattan, Hays and at a satellite site in Colby, Kansas to test the limits of tolerance and range extension into drier areas). In this experimental design, we have grown in 4 climate locations plants of 3 ecotypes and a commonly used cultivar Kaw in replicated blocks (n=10) with each plant growing singly in plastic and in artificially assembled seeded communities (n=4 per planting location, 4x4 m plots). In summer 2012, we focused on measuring plant, community, and ecosystem response to experimental drought. We measured leaf-level gas exchange and photosynthesis and water potential 3 times in 2012 (once each summer month), determined ecotypic-specific plant cover mid-season to assess ecotype specific cover response to altered rainfall, harvested plots for vegetative and seed production (ANPP), identified phenological differences among ecotypes, measured soil moisture throughout the season, monitored precipitation. Collected leaf samples from all sites for transcriptome sequencing and phytohormones in ambient and sheltered plots. Began collaboration with Dr Sue Brown and the KSU Bioinformatics Center in the exploration and development of a de novo reference transcriptome hybrid assembled from 454 (2 runs) and Illumina reads (600 million). The hybrid-assembled transcriptome will be a critical resource for future mapping of Illumina reads from drought and control samples. Beginning to expand our markers using GBS with Dr Jesse Poland. 28 Presentations: Developed a bluestem ecotype website: www.bluestemecotype.ksu.edu to facilitate dissemination about the project and to encourage collaboration.USDA-Manhattan PMC uses this reciprocal garden Manhattan site as a demonstration site on the role of ecotypic variation in big bluestem, Richard Wynia is involved as a collaborator. PARTICIPANTS: Eduard Akhunov-Plant Pathology-KSU expertise genomics Paul St Amand, USDA Genotyping Lab-assisting with AFLP analysis Karen Garrett-Big Bluestem ecology and genomics-Plant Pathology Ted Morgan-Biology-KSU-expertise in population genetics Keith Harmoney- Range Scientist, KSU W KS Agricultural Res Center, Hays KS Sara Baer-SIU Plant Biology, expertise in restoration ecology, 2 students supported Brian Maricle-Biology (FHSU) new subcontract, grad student Jake Olsen supported Students Trained KSU Undergrads:Lauren Wheeler (2010), Meghan Kelly, Cody Rodewald, Angel DeLaCruz Jessica Gaffney (summer SUROP student 2010, 2011, 2012 respectively), Sofia Sabates (2011-2012 URM), Amber Jensen (2012 REU), Mathew Gaillart and Chadd Harrison (both 2011-2012) SIU Undergrads: Evan Mcrea SIU, Garret Sendor FHSU: KS, Keri Caudle (winner of prestigious national undergrad fellowships from ASPB, EPA, BSA Undergrad Research Student of the Year 2012) Training: Five graduate students (2 from KSU, 2 from SIU, 1 from FHSU) have successfully defended their research based on their reciprocal garden work. Three students are in progress. Graduate students* defended: Miranda Gray*, Hannah Tetreault, Jennifer Shelton*, Janneke Ravenek(Amsterdam exchange MS student), Jake Olsen* (FHSU biology student) Katelyn ONeil (FHSU), Rachel Goad* SIU, Meredith Mendola SIU, Juliette Donatelli SIU, Laurel Wilson SIU, Gin Kan* and K Zhang*, Biological and Agricultural Engineering KSU. One of the students (Miranda Gray) was instrumental in attracting Sigma Xi and Plants for the Heartland funding. New Participants Dr. Jesse Poland, USDA-ARS-KSU Dr. Sue Brown, KSU Bioinformatics Center Drs. Ruth Welti, Richard Jeannotte, Thilani Samarakoon, KSU Lipidomics Center Drs. Basil Nikolau, Zhihong Song, Keck Metabolmic Center and Biochemistry, Iowa State University-lipidome of bluestem Dr. Carolyn Malmstrom, University of Wisconsin Madison-ecotypic differences in virus infection and resistance Drs. Anna Whitfield, Dorith Rottenburg, KSU Plant Pathology, ecotypic differences in virus infection and resistance Dr. Nora Bello-KSU Statistics, with expertise in quantitative statistical genetics Dr. Kevin Price and student Nan An KSU Agronomy and Remote Sensing, Remote sensing of plant performance and phenology of bluestem ecotypes Drs Donghai Wang, Wenqiao Yuan, Richard Nelson-KSU Biological and Agricultural Engineering KSU-Biofuel potential of bluestem ecotypes TARGET AUDIENCES: Constructing rainout shelters for all ecotypes at all sites, instead of shelters for resident ecotypes only --Sequencing bluestem drought transcriptome using a combination of 454 and Illumina reads. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Genomic Tool Development. We are continuing to develop a new genomic tools for big bluestem Big bluestem, a close relative of maize, has a large, complex polyploid genome. With new genomics approaches, however, these challenging genomes are becoming more tractable. We genotyped 378 plants from 11 populations from three precipitation regions (central and eastern KS, Illinois), resulting in 387 AFLP markers. Structure analyses supported six genetic clusters, with Kansas populations belonging to three genetic groups, and Illinois populations having allele frequencies divergent from Kansas populations. Eight FST outlier loci under diversifying selection were linked to precipitation. A new Genotyping by Sequencing approach has been shown to be robust across species and produces thousands of molecular markers. We are collaboration with Poland in the development of GBS tools for big bluestem. We are also developing de novo transcriptome resources for the two most divergent ecotypes (xeric and mesic). This resource is a valuable tool to identify transcriptional differences and SNPs associated with drought tolerance traits. We used 454 sequencing and Newbler de novo transcriptome assembly, complemented with additional Illumina RNA-sequencing. Studies on phenotypic and genetic variation will guide the development of germplasm that is locally adapted to drought. We received three seed grants to defray costs of sequencing. Field Experiment-Reciprocal Garden Transplant Studies. Reciprocal transplant common gardens allow us to test for genotype by environment interaction and to detect evidence of adaptive differentiation across the precipitation gradient at the 4 sites spanning from Colby KS to Carbondale Illinois. Plant establishment data in the reciprocal gardens from Oct 2009-2012 suggest local ecotypic adaptation. At the western sites, in Hays and Colby, KS, the central KS ecotype was represented by a significantly greater numbers of plants than the other ecotypes, which suggests adaptive differentiation of the central KS ecotype to low precipitation. Field data from summers 2010-2012 demonstrate a continued strong phenotypic cline in drought tolerance of the three ecotypes (Hays, Manhattan, and Illinois) in both the seeded communities and singly grown plants. The westernmost ecotype, exhibits drought-adapted features (dwarf stature, thicker leaves, reduced canopy area) compared to the other ecotypes. In the driest planting location (Colby) the westernmost Hays ecotype has 3x the plant cover relative to the other ecotypes, thus suggesting local drought adaptation. Interestingly, phenology is ecotype-specific with the Hays ecotype, wherever it is planted, is always about one month advanced in growth and development relative to the Illinois ecotype, thus escaping drought in late summer. We are currently analyzing data on the experimental drought response of the ecotypes across the gradient in terms of gas exchange, WUE, and water potential. We are developing the drought de novo reference transcriptome for future, planned gene expression studies in replicated rain out and control plots of the ecotypes across the reciprocal gardens.
Publications
- Gan Jing, Wenqiao Yuan, Loretta Johnson, Donghai Wang, Richard Nelson, Ke Zhang. Hydrothermal conversion of big bluestem for bio-oil production: The effect of ecotype and planting location. Presentation, DOT SunGrant Conference, Oct 2012. New Orleans
- Olsen, J.T., H.M. Tetreault, R.K. Goad, M.L. Mendola, L.C. Johnson, S.G. Baer, and B.R. Maricle. 2011. Variation of photosynthesis in big bluestem and sand bluestem reciprocally planted across a precipitation gradient. Transactions of the Kansas Academy of Science 114:
- Zhang, Ke Loretta Johnson, Richard Nelson, Wenqiao Yuan, Zhijian. Pei, and Donghai Wang. 2012. Chemical and elemental composition of big bluestem as affected by ecotype and planting location along the precipitation gradient of the Great Plains. Industrial Crops and Products 40:210.
- Goad R.K., H.M. Tetreault, K. Elsenbroek, D. Carpenter, J.T. Olsen, B.R. Maricle, L.C. Johnson, and S.G. Baer. Patterns in aboveground net primary productivity in prairie reciprocally restored with dominant grasses from across a precipitation gradient. Presented at the Ecological Society of America Annual Meeting, Austin, TX, August 2011 (poster presentation)
- Tetreault, H., C. Rodewald, S. Baer, B. Maricle, T. Morgan, R. Goad, J. Olsen, and L. Johnson. Local drought adaptation of the ecologically dominant prairie grass big bluestem Andropogon gerardii: Contribution of genotype and environment to phenotypic variation. Present at the Ecological Society of America Annual Meeting, Austin, TX, August 2011 (poster presentation)
- Shelton, Jennifer M, Ravenek Janneke, Samarakoon Thilani, Jeannotte Richard, Welti Ruth, Nikolau, Basil J, Song Zhihong, and Johnson, Loretta. Divergence in leaf morphology and physiology in Andropogon gerardii var. hallii (sand bluestem) a locally adapted variety of Andropogon gerardii (big bluestem) Presented ASPB, Minneapolis, Aug 2011 (poster presentation)
- Loretta Johnson, Invited lecture for workshop on Ecological Genomics, Mount Desert Island Biological Laboratory, Sept 2011.
- Gray, Miranda1; Tetreault, Hannah; Rodewald, Cody; St. Amand, Paul; Baer, Sara; Maricle, Brian; Morgan, Ted; Bello, Nora Goad, Rache; Olsen, Jake; Wheeler, Lauren; Akhunov, Eduard; Garrett, Karen; Bai, Guihua, and Johnson, Loretta Evidence of Local Adaptation and Genetic Differentiation across Andropogon gerardii (big bluestem) Tall Grass Ecotypes Occurring along a Precipitation Gradient. Poster, Ecological Genomics Symposium, Nov 4-6, 2011, Kansas City.
- Johnny Bryant, Angel DeLa Cruz, Kevin Price, Nan An, Mary Knapp, Hannah Tetreault, Brian Maricle, Jake Olsen, Keri Caudle, Sara Baer, Loretta Johnson. Remote sensing to detect plant performance in common gardens of big bluestem ecotypes across the Great Plains. Poster, Ecological Genomics Symposium, Nov 4-6, 2011, Kansas City.
- Loretta Johnson. Phenotypic and genetic variation in an ecologically dominant grass Andropogon gerardii along the Great Plains precipitation gradient: A reciprocal garden approach Invited Speaker, Plant Pathology Department, KSU, Feb 2012.
- Gray, Paul St. Amand, Loretta Johnson, Eduard Akhunov, Karen Garrett, Mary Knapp, Alina Akhunova, Sanjay Chellapilla, Ted Morgan, Guihua Bai. Genetic differentiation of Andropogon gerardii(big bluestem) ecotypes along a natural precipitation gradient Miranda Botanical Society of America. Columbus July 2012 (oral Presentation)
- KSU Graduate School SUROP (Summer Undergraduate Research Opportunities) seminar, July, 2012. Jessica Gaffney, Variations in polyploidy in Big Bluestem (Andropogon gerardii) ecotypes across the Great Plains
- KSU Graduate School REU (Summer Undergraduate Research Opportunities) seminar, July, 2012. Amber Jensen, Big bluestem ecotypic differences in leaf nitrogen content as a possible mechanism for increased performance of western Kansas ecotypes in reciprocal gardens planted across the Great Plains. Amber will continue this research for her Senior Thesis, at Nebraska Welsyean.
- Ecotypic variation in drought tolerance and genetic diversity of an ecologically dominant grass big bluestem (Andropogon gerardii) across the Great Plains precipitation gradient Gray, Miranda, St Amand Paul, Tetreault, Hannah; Baer, Sara; Maricle, Brian; Morgan, Ted; Goad, Rachel; Olsen, Jake and Johnson, Loretta. Ecological Society of America. Portland Oregon, Aug. 2012.
- Meredith L Mendola, Sara G Baer, Loretta C Johnson. Ecotype and climate controls on belowground ecosystem processes. Oral Presentation. Ecological Society of America. Portland Oregon, Aug. 2012.
- Zhang, Ke Loretta Johnson, Richard Nelson, Wenqiao Yuan, Zhijian. Pei, and Donghai Wang. Chemical and elemental composition of big bluestem as affected by ecotype and planting location along the precipitation gradient of the Great Plains. Presentation, DOT SunGrant Conference, Oct 2012. New Orleans
- Loretta Johnson. Conservation Genetics of an Ecologically Dominant Grass of the Great Plains. Invited Speaker, EU Workshop and Advisor to the Conservation Genomics Consortium, Seville-Spain, Feb 2011.
- All Participant Ecotype Project Workshop. Held all-participant meeting to discuss USDA project coordination, data analyses, manuscript submissions. April, 2012
- Olsen, J.T., H.M. Tetreault, R.K. Goad, M.L. Mendola L.C. Johnson, S.G. Baer, and B.R. Maricle. Variation of photosynthesis in big bluestem and sand bluestem reciprocally planted across a precipitation gradient. Presented at the Kansas Academy of Science Annual Meeting, Baldwin City, KS, April 2012 (oral presentation)
- Miranda Gray, Paul St. Amand, Loretta Johnson, Eduard Akhunov, Karen Garrett, Alina Akhunova, Ted Morgan, Guihua Bai. Employing a whole-genome scan to differentiate Andropogon gerardii (big bluestem) ecotypes ASPB, Poster Presentation, Lincoln Nebraska, March 2012. Gray received the top poster award!
- Loretta Johnson, Invited Speaker, Ecological Genomics Symposium, Oct 26-28, 2012, KC.Phenotypic and genetic variation in an ecologically dominant grass Andropogon gerardii along the Great Plains precipitation gradient: A reciprocal garden approach.
- Thilani N. Samarakoon, Jennifer M. Shelton, Richard Jeannotte, Zhihong Song, Nora Bello, Basil Nikolau, Ruth Welti, Loretta Johnson. A Gas Chromatography/Mass Spectrometry Reveals Contrasts in Epicuticular Wax Composition of Big Bluestem (Andropogon gerardii) and edaphic variety Sand Bluestem (A. gerardii var halli.)ASPB, Oral Presentation, Lincoln Nebraska, March 2012.
- Shelton, Jennifer M, Ravenek Janneke, Samarakoon Thilani, Jeannotte Richard, Welti Ruth, Nikolau, Basil J, Song Zhihong, and Johnson, Loretta. Divergence in leaf morphology and physiology in Andropogon gerardii var. hallii (sand bluestem) a locally adapted variety of Andropogon gerardii (big bluestem) ASPB, Poster Presentation, Lincoln Nebraska, March 2012.
- Olsen, J.T., H.M. Tetrault, R.K. Goad, M.L. Mendola, L.C. Johnson, S.G. Baer, and B.R. Maricle. Photosynthetic variation of big bluestem and sand bluestem influenced by ecotype and precipitation. Presented at the Botanical Society of America Annual Meeting, Columbus July 2012 (oral presentation)
- Gray, Miranda, St Amand Paul, Tetreault, Hannah; Baer, Sara; Maricle, Brian; Morgan, Ted; Goad, Rachel Olsen, Jake and Johnson, Loretta Phenotypic and genetic variation in a dominant prairie grass along the Great Plains precipitation gradient: a reciprocal garden approach. Poster, Ecological Genomics Symposium, Oct 26-28, 2012, KC.
- Gaffney, Jessica; Sabates, Sofia; Baer, Sara; Maricle, Brian; McCrea, Evan; Bello, Nora M. and Johnson, Loretta Variations in polyploidy in Big Bluestem (Andropogon gerardii) ecotypes across the Great Plains. Poster, Ecological Genomics Symposium, Oct 26-28, 2012, KC.
- Sabates, Sofia; Baer, Sara; Caudle, Keri; Harmoney, Keith; Maricle, Brian; McCrea, Evan; and Johnson, Loretta. Phenology and seed production in Big Bluestem (Andropogon gerardii) ecotypes in Great Plains reciprocal gardens: the role of site and ecotype Poster, Ecological Genomics Symposium, Oct 26-28, 2012, KC.
- Loretta C Johnson, Miranda Gray, Alina Akhunova, Susan J. Brown, Sanjay Chellapilla, Theodore Morgan, Karen Garrett, Hanquan Liang and Eduard Akhunov, Loretta Phenotypic and genetic variation in a dominant forage and biofuel grass along the Great Plains precipitation gradient: a reciprocal garden approach. To be presented at the 2013 PAG, San Diego.
- Shelton, Jennifer, Miranda M. Gray, Susan J. Brown, Sanjay Chellapilla, Alina R. Akhunova, Eduard D. Akhunov, Hanquan Liang, Loretta C. Johnson De novo transcriptome profiling of two edaphically and phenotypically divergent grasses: dominant forage grass big bluestem Andropogon gerardii and drought-tolerant sand bluestem Andropogon gerardii ssp. Hallii. To be presented at the 2013 PAG, San Diego.
- Gan Jing, Wenqiao Yuan, Loretta Johnson, Donghai Wang a, Richard Nelson, Ke Zhang. 2012. Hydrothermal conversion of big bluestem for bio-oil production: The effect of ecotype and planting location. Bioresource Technology 116:413.
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Progress 09/01/10 to 08/31/11
Outputs
OUTPUTS: In Spring 2011, we continued the second year of measurements on the reciprocal common gardens of bluestem ecotypes. The gardens were established in each of the precipitation zones of the Great Plains to measure genotype by environment interaction and to detect evidence of adaptive differentiation across the precipitation gradient at three sites (Carbondale Illinois, Manhattan, Hays and at a satellite site in Colby, Kansas to test the limits of tolerance and range extension into drier areas). In this experimental design, we have grown in 4 climate locations plants of 3 ecotypes (each comprised of 4 separate populations) and a commonly used cultivar Kaw in replicated blocks (n=10) with each plant growing singly in plastic and in artificially assembled seeded communities (n=4 per planting location, 4x4 m plots). The major activity for 2011 was the assembly and construction of rainout shelters for the 3 main study sites in Carbondale, Manhattan, and Hays. These shelters are lexan panels that channel 50% of the rainfall into gutters and collected in rain barrels. The shelters follow the design of Sala in terms of construction. Our shelters are about 2.5 m x 2.5 m, cover all ecotypes in the 3 locations, and are paired with ambient rainfall plots (split plot). Thus, each site has a total of 12 shelters, and a total for the experiment of 36. We also established a soil moisture monitoring protocol to document soil moisture differences between shelter and ambient as well as a constellation of rain gauges in sheltered and ambient plots. The shelters were established in the first week of June in all sites. In summer 2011, we focused mainly on measuring plant, community, and ecosystem response to experimental rainout drought. We made the following functional, morphological and performance responses of the bluestem ecotypes in all four planting locations in the seeded reciprocal gardens: Measured leaf-level gas exchange and photosynthesis 3 times in 2011 (once each summer month), determined ecotypic-specific plant cover in June and August 2011 to assess ecotype specific establishment and survival in response to altered rainfall, harvested ANPP well as determined canopy LAI, identified phenological differences among ecotypes, collected in Aug. leaf samples from all sites for transcriptome sequencing (454), measured soil moisture throughout the season, monitored precipitation. Based on data from 2010 showing intrinsic phenological differences among ecotypes, we pursued this further by assessing changes in plant performance and senescence with a SPAD 502 chlorophyll meter and more fine scale resolution of spectral reflectance (2500 bands) with a spectral radiometer in collaboration with doctoral student Nan An and Dr Kevin Price in remote sensing lab. In collaboration with Drs. Malmstrom, Whitfield, and Rottenburg, assessed viral infection and resistance differences among ecotypes. Collected in Aug. leaf samples from all sites for transcriptome sequencing (454) and phytohormones in ambient and sheltered plots, Continued to develop AFLP markers from 24 plants per population, 12 populations comprised of 3 ecotypes. PARTICIPANTS: Eduard Akhunov-Plant Pathology-KSU expertise genomics Paul St Amand, USDA Genotyping Lab-assisting with AFLP analysis Karen Garrett-Big Bluestem ecology and genomics-Plant Pathology Ted Morgan-Biology-KSU-expertise in population genetics, Keith Harmoney- Range Scientist, KSU W KS Agricultural Res Center, Hays KS, Sara Baer-SIU Plant Biology, expertise in restoration ecology, 2 students supported Brian Maricle-Biology (FHSU) new subcontract, grad student Jake Olsen supported. Undergrads Lauren Wheeler, Meghan Kelly, Cody Rodewald, Angel DeLaCruz summer SUROP student 2010, 2011 respectively), Garret Sendor SIU, Keri Caudel (FHSU), Evan Mcrea SIU. Graduate students: Miranda Gray, Hannah Tetreault, Jennifer Shelton, Janneke Ravenek(Amsterdam exchange MS student), Jake Olsen (FHSU biology student), Rachel Goad SIU, Meredith Mendola SIU, Juliette Donatelli SIU, Gin Kan and K Zhang, Biological and Agricultural Engineering KSU. Drs. Ruth Welti, Richard Jeannotte, Thilani Samarakoon, KSU Lipidomics Center, Drs. Basil Nikolau, Zhihong Song, Keck Metabolmic Center and Biochemistry, Iowa State University-lipidome of bluestem, Dr. Carolyn Malmstrom, University of Wisconsin Madison-ecotypic differences in virus infection and resistance Drs. Anna Whitfield, Dorith Rottenburg, KSU Plant Pathology, ecoytpic differences in virus infection and resistance, Dr. Nora Bello-KSU Statistics, with expertise in quantitative statistical genetics Dr. Kevin Price and student Nan An KSU Agronomy and Remote Sensing, Remote sensing of plant performance and phenology of bluestem ecotypes, Drs Donghai Wang, Wenqiao Yuan, Richard Nelson-KSU Biological and Agricultural Engineering KSU-Biofuel potential of bluestem ecotypes. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Constructing rainout shelters for all ecotypes at all sites, instead of shelters for resident ecotypes only. Sequencing bluestem drought transcriptome and making custom big bluestem chip for use in gene expression studies summer 2010, instead of using cross-species hybridization with Zea chip
Impacts
Reciprocal transplant common gardens allow us to test for genotype by environment interaction and to detect evidence of adaptive differentiation across the precipitation gradient. Plant establishment data in the reciprocal gardens already suggest local ecotypic adaptation. At the western sites, in Hays and Colby, KS, the Hays ecotype was represented by a significantly greater numbers of plants than the other ecotypes, which suggests adaptive differentiation of the Hays ecotype to areas of low precipitation. Field data from summers 2010-2011 demonstrate a strong phenotypic cline in drought tolerance of the three ecotypes (Hays, Manhattan, and Illinois) in both the seeded communities and singly grown plants. The westernmost ecotype, (Hays) exhibits drought-adapted features (dwarf stature, thicker leaves, reduced canopy area) compared to the other ecotypes. Our data demonstrate strong planting site and ecotype effects as well as interaction between ecotype and planting site (G X E). In the driest planting location (Colby) the westernmost Hays ecotype has 3x the plant cover relative to the other ecotypes, thus suggesting local drought adaptation. In August 2010 and 2011, we collected samples for ANPP to detect if plot level ANPP is affected by ecotypes across the 4 planting locations and by experimental drought. Interestingly, phenology is ecotype-specific with the Hays ecotype, wherever it is planted, is always about one month advanced in growth and development relative to the Illinois ecotype, thus escaping drought in late summer. We found the Hays ecotype had the highest chorophyll spectral absorbance among ecotypes corroborating the high photosynthetic rates in that ecotype. New studies on pathogen sorting among ecotypes showed the Hays ecotype has the highest incidence of rust infection, independent of planting location. We are currently analyzing data on the experimental drought response of the ecotypes across the gradient. We are continuing to develop a new set of genomic tools for big bluestem, instead of relying on the maize cross species hybridization and associated difficulties. We have collected leaves of the bluestem ecotypes from seeded reciprocal gardens and under experimental drought for mRNA isolation and will construct enriched cDNA libraries to be analyzed for gene expression using the Roche 454 Genome Sequencer. We are now doing a 454 sequencing run on cDNA libraries from experimentally droughted big bluestem to detect differential response of the ecotypes and identification of candidate genes. Lipidomics. Through several pilot studies using the GC-MS in the KSU Lipidomics Center and others, we are detecting very distinct lipid profiles among bluestem ecotypes. Gas chromatography mass spectrometry (GC-MS) indicates that big bluestem and sand bluestem are ecotypically quite distinctive in both epicuticular wax load and composition. The sand bluestem accession had more than five times as much epicuticular wax per leaf dry weight than the big bluestem cultivar and was characterized by a high content of diketones. The high diketone was also corroborated by the presence of tubules in associated SEM photos (2K magnification).
Publications
- Phenotypic cline in big bluestem ecotypes: Contribution of genotype and environment to phenotype using a reciprocal garden approach. Tetreault, H , M. Gray, C. Rodewald, P. St. Amand, G. Bai, E. Akhunov, K. Garrett, S. Baer, B. Maricle, T. Morgan, N. Bello, R. Goad, J. Olsen, and L. Johnson. In prep. 2011.
- Differential establishment, cover, and productivity of big bluestem ecotypes seeded in reciprocal transplant gardens across the Great Plains: Evidence for local adaptation to drought. Hannah Tetreault, Loretta Johnson, Racheal Goad, Sara Baer, B.R. Maricle, Jake Olsen, In prep. 2011.
- Ecotype-specific Leaf-level differences in photosynthesis of big bluestem ecotypes in reciprocal transplant gardens across the Great Plains-B.R. Maricle, Jake Olsen, Loretta Johnson, Hannah Tetreault, Sara Baer, Racheal Goad. In prep. 2011.
- Local adaptation of cuticular wax biosynthesis in big bluestem Andropogon gerardii and sand bluestem Andropgon gerardii var hallii. Jennifer M. Shelton, Richard Jeannotte, Loretta C. Johnson, Zhihong Song, Basil Nickolau, Ruth Welti. In prep. 2011.
- Olsen, J.T., H.M. Tetreault, R.K. Goad, M.L. Mendola, L.C. Johnson, S.G. Baer, and B.R. Maricle. 2011. Variation of photosynthesis in big bluestem and sand bluestem reciprocally planted across a precipitation gradient. Transactions of the Kansas Academy of Science 114:
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Progress 09/01/09 to 08/31/10
Outputs
OUTPUTS: In Spring 2009, reciprocal common gardens were established in each of the precipitation zones to measure genotype by environment interaction and to detect evidence of adaptive differentiation across the precipitation gradient at three sites (Carbondale Illinois, Manhattan, Hays and at a satellite site in Colby, Kansas to test the limits of tolerance and range extension into drier areas). In this experimental design, we have grown in 4 climate locations plants of 3 ecotypes (each comprised of 4 separate populations) and a commonly used cultivar Kaw in replicated blocks (n=10) with each plant growing singly in plastic and in artificially assembled seeded communities (n=4 per planting location, 4x4 m plots). Rainout shelters will be constructed and put in place spring 2011. We made the following functional, morphological and performance responses of the bluestem ecotypes in all four planting locations in the seeded reciprocal gardens: Measured leaf-level gas exchange and photosynthesis 3 times in 2010 (once each summer month), determined ecotypic-specific plant cover in June and August 2010 to assess ecotype specific establishment and survival across the precipitation gradient, harvested ANPP well as determined canopy LAI, identified phenological differences among ecotypes, collected in Aug. leaf samples from all sites for transcriptome sequencing (454), measured soil moisture throughout the season, Monitored precipitation. We made the following functional, morphological and performance responses of the bluestem ecotypes in all four locations in single ecotype plantings (12 populations, 4 per ecotype 10 blocks) at each site to measure individual plant responses: Measured leaf-level gas exchange and photosynthesis 3 times in 2010 (once each summer month), Determined canopy height, canopy area of single plants 2 times in the season, blade width, Harvested single plants to estimate ending mass differences, Identified phenological differences among ecotypes, Determined differences in rust infection, An additional 10 blocks (already in place) will also receive a rain out treatment at all sites next spring/summer 2011. Continued to develop AFLP markers from 24 plants per population, 12 populations comprised of 3 ecotypes. Collected in Aug. leaf samples from all sites for transcriptome sequencing (454) and phytohormones, plan to analyze bluestem seed from additional populations in order to place our limited 12 populations (4 pop per ecotype) into a broader genetic and ecological context to test for genetic structure of big bluestem across the Great Plains. Ecological Genomics Symposium, Nov 5-7, 2010, Marriot Hotel, Kansas City, Contributed poster, Local adaptation of wax biosynthesis in big bluestem Andropogon gerardii and sand bluestem Andropogon gerardii var hallii. Jennifer Shelton, Loretta Johnson, Janneke Ravenek, Richard Jeanotte, Zhihong Song, Ruth Welti. Ecology and Evolutionary Biology seminar, KSU. September 2010. PARTICIPANTS: Eduard Akhunov Plant Pathology KSU expertise genomics Paul St Amand, USDA Genotyping Lab assisting with AFLP analysis Karen Garrett-Big Bluestem ecology and genomics Plant Pathology Ted Morgan Biology KSU expertise in population genetics Keith Harmoney Range Scientist, KSU W KS Agricultural Res Center, Hays KS Sara Baer SIU Plant Biology, expertise in restoration ecology, 2 students supported Brian Maricle-Biology (FHSU) new subcontract, grad student Jake Olsen supported Students Trained Undergrads Lauren Wheeler, Meghan Kelly, Cody Rodewald (summer SUROP student) Graduate students: Miranda Gray, Hannah Tetreault, Jennifer Shelton, Janneke Ravenek(Amsterdam exchange MS student), Jake Olsen (FHSU biology student) New Participants (see more info info below under modifications)Richard Wynia, USDA Plant Materials Center, Manhattan KS Danny Foster-KSU W KS Agricultural Res Center, Colby KS Mary Knapp, KS State Climatologist Brian Maricle-Biologist, Fort Hays S. Univ. Richard Jeannotte, KSU Lipidomics Center Chris Pires/Roxi Sttel, U. Mo. Columbia, TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Sequencing bluestem drought transcriptome and making custom big bluestem chip for use in gene expression studies summer 2010, instead of using cross-species hybridization with Zea chip--Added Brian Maricle, Asst Professor in Biology at Fort Hays State University, as a collaborator. This collaboration will provide training opportunities for MS level students in Hays.
Impacts
Field Experiment-Reciprocal Garden Transplant Studies. Data from the reciprocal gardens already suggest adaptive differentiation of the Hays ecotype to areas of low precipitation. Field data from the summer 2010 corroborate a strong phenotypic cline in drought tolerance of the three ecotypes (Hays, Manhattan, and Illinois) in seeded communities and singly grown plants. The westernmost ecotype, (Hays) exhibits drought-adapted features (dwarf stature, thicker leaves, reduced canopy area) compared to the other ecotypes. Our data demonstrate strong planting site and ecotype effects as well as interaction between ecotype and planting site. Furthermore, we estimated plant cover of the three ecotypes across the 4 planting sites in our seeded artificially assembled communities. In the driest planting location (Colby) the westernmost Hays ecotype has 3x the cover relative to the other ecotypes, thus suggesting local drought adaptation. Interestingly, phenology is ecotype-specific with the Hays ecotype, wherever it is planted, is always about one month advanced relative to the Illinois ecotype, thus escaping drought in late summer. Unexpectedly, we found the Hays ecotype had the highest photosynthetic rates among ecotypes as well as the highest incidence of rust infection, independent of planting location. Genomic/Metabolomic Tool Development. We are continuing to develop a new set of genomic tools for big bluestem using 454 sequencing. We have collected leaves of the bluestem ecotypes from seeded reciprocal gardens for mRNA isolation and will construct enriched cDNA libraries to be analyzed for gene expression using the Roche 454 Genome Sequencer. A 454 sequencing run on cDNA libraries from droughted big bluestem will detect differential response of the ecotypes and help identify candidate genes. Began new collaborations with Dr. Richard Jeannotte from the KSU Lipidomics Center to analyze lipid profiles of bluestem ecotypes, with focus on the waxy epicuticle as a drought adaptation. Began collaborations with Chris Pires/Roxie Steel (Univ. of MO, Columbia) on Illumina sequencing of chloroplast genome of several different ecotypes. Received three seed grants (total $20,400) to help defray costs of sequencing. One of the students (Miranda Gray) was instrumental in attracting Sigma Xi funding to provide support for reagents. Lipidomics. Through several pilot studies using the GC-MS in the KSU Lipidomics Center and collaborations with Drs. Richard Jeannotte, we are detecting very distinct lipid profiles among bluestem ecotypes. GC-MS profiles indicate that big bluestem and sand bluestem are quite distinctive in both epicuticular wax load and composition. The sand bluestem accession had more than five times as much epicuticular wax per leaf dry weight than the big bluestem cultivar and was characterized by a high content of diketones. The high diketone was also corroborated by the presence of tubules in associated SEM photos (2K magnification). Jennifer Shelton, a new graduate student on the project, is following up on this important finding by analyzing a full suite of bluestem ecotypes using GC-MS as well as DART.
Publications
- Phenotypic cline in big bluestem ecotypes: Contribution of genotype and environment to phenotype using a reciprocal garden approach. Hannah Tetreault, Cody Rodewald, Loretta Johnson, Racheal Gode, Sara Baer, B.R. Maricle, Jake Olsen, In prep. To be submitted mar 2011.
- Differential establishment, cover, and productivity of big bluestem ecotypes seeded in reciprocal transplant gardens across the Great Plains: Evidence for local adaptation to drought. Hannah Tetreault, Loretta Johnson, Racheal Gode, Sara Baer, B.R. Maricle, Jake Olsen, In prep. To be submitted mar 2011.
- Big bluestem ecotypic differences in rust infection in reciprocal transplant garden across the Great Plains-K Garrett, L. Johnson et al. (In prep, To be submitted mar 2011)
- Ecotype-specific Leaf-level differences in photosynthesis of big bluestem ecotypes in reciprocal transplant gardens across the Great Plains-B.R. Maricle, Jake Olsen, Loretta Johnson, Hannah Tetreault, Sara Baer, Racheal Gode. In prep. To be submitted mar 2011.
- Divergent phenotypes in sand bluestem and big bluestem ecotypes of the Great Plains: Local adaptation to edaphic environment. Janneke M. Ravenek, Jennifer M. Shelton, Richard Jeannotte, Zhihong Song , Basil Nickolau, Loretta C. Johnson. In prep. To be submitted Jan 2011.
- Local adaptation of cuticular wax biosynthesis in big bluestem Andropogon gerardii and sand bluestem Andropgon gerardii var hallii. Jennifer M. Shelton , Richard Jeannotte, Loretta C. Johnson, Zhihong Song, Basil Nickolau, Ruth Welti. In prep. To be submitted June 2011.
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Progress 09/01/08 to 08/31/09
Outputs
OUTPUTS: Activities: Collected big bluestem seed (20kg) from 12 populations in 3 geographic areas (Hays, Manhattan, Carbondale) in fall 2008 to be used in reciprocal garden ecotype transplants of the bluestem ecotypes. Tested for seed germination and viability. Established in summer 2009 reciprocal garden transplants at the following field locations-Agricultural Experiment station in Carbondale, the USDA Plant Material Center in Manhattan, KSU agricultural research center in Hays, Kansas and Colby, Kansas. Each site has 4 replicated blocks, containing each ecotype, all three ecotypes mixed, and a commonly used CRP Bluestem cultivar Kaw. Rainout shelters will be constructed and put in place for spring 2010. Measured soil moisture throughout the season in all 4 sites, Monitored precipitation in all 4 sites, In September 2009, measured establishment (numbers) of big bluestem plants in all plots in all 4 locations. Established plantings (10 blocks) of big bluestem individuals of each ecotype into shade cloth at each site to measure individual plant responses to precipitation along the Great Plains climatic gradient. An additional 10 blocks (already in place) will also receive a rain out treatment at all sites next spring/summer 2010. Collected additional bluestem seed from additional populations in order to place out limited 12 populations (4 pop per ecotype) into a broader genetic and ecological context to test for genetic structure of big bluestem across the Great Plains. So far we have seeds from about 30 additional populations spanning the extremes of big bluestem distribution from CO, OK, ND and IL. Began AFLP studies of populations. Events: Ecological Genomics Symposium, Nov 14-16, 2008, InterContinental Hotel, Kansas City, Contributed poster, Ecotypic Variation in Big Bluestem Andropogon gerardii across a precipiation gradient in Midwest Grasslands. Bengtson, B.A. Ecotypic Variation and Drought Tolerance of Andropogon gerardii Across a Precipitation Gradient in Midwest Grasslands. Presentation for Kansas Research Forum at Kansas State University, Manhattan, KS. 6 March, 2009, and Presentation for the 35th Annual Forum for Student Research, Kansas State University, Manhattan, KS. 7 March, 2009. Nathan Spaht et al. Occurrence and distribution of big bluestem polyploids in tall grass prairie: Ecological consequences of genome size. Presentation for Kansas Research Forum at Kansas State University, Manhattan, KS. 6 March, 2009. Konza LTER workshop, Mar 28, 2009. Ecological Genomic of Prairie Grasses. Gordon Conference 2009-Ecolgocial and Evolutionary Functional genomics. July 7-12, Tilton, NH. American Genetics Association Symposium: Genetics and Genomics of Environmental Change, June 8-11, Brown University. All Participant Ecotype Project Workshop. Held 2 all-participant meetings to discuss USDA project design etc and begin implementation of the project. Dissemination: USDA-Manhattan PMC uses this reciprocal garden Manhattan site as a demonstration site on the role of ecotypic variation in big bluestem, Richard Wynia is involved as a collaborator. PARTICIPANTS: Loretta Johnson, Project Director; Eduard Akhunov-Plant Pathology-KSUexpertise genomics; Paul St. Amand, USDA Genotyping Lab-assisting with AFLP analysis; Karen Garrett-Big Bluestem ecology and genomics-Plant Pathology; Ted Morgan-Biology-KSU-expertise in population genetics; Keith Harmoney- Range Scientist, KSU W KS Agricultural Res Center, Hays KS; and Sara Baer-SIU Plant Biology, expertise in restoration ecology. Students Trained: Undergrads - Kate Zimmerman, Nathan Spath (both conducted indepenet res projects); Graduate students: Bethany Begston, Miranda Gray, and High School student-Jordan Smith. New Participants (see more info info below under modifications): Richard Wynia, USDA Plant Materials Center, Manhattan KS; Danny Foster-KSU W KS Agricultural Res Center, Colby KS; Mary Knapp, KS State Climatologist; and Brian Maricle-Biologist, Fort Hays State University. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Sequencing bluestem transcriptome and making custom big bluestem chip for use in gene expression studies summer 2010, instead of using cross-species hybridization with Zea chip. Added Colby Kansas site located at the W KS Agricultural Research Center as a reciprocal garden site to test the limits of drought tolerance in the westernmost part of the range of big bluestem. Added Brian Maricle, Asst Professor in Biology at Fort Hays State University, as a collaborator. This collaboration will provide training opportunities for MS level students in Hays. Added the USDA Plant Materials Center (Richard Wynia, Director) as one of our reciprocal garden site in Manhattan, instead of using Konza Prairie. Added Mary Knapp, State climatologist, as a collaborator. May has been very helpful in helping access and analyze weather records across the Great Plains in order to put our experimental plots into a climate contest. Collected additional bluestem seed from additional populations in order to place out limited 12 populations (4 pop per ecotype) into a broader genetic and ecological context to test for genetic structure of big bluestem across the Great Plains. So far we have seeds from about 30 additional populations spanning the extremes of big bluestem distribution from CO, OK, ND and IL. Began AFLP studies of populations. Established plantings (10 blocks) of big bluestem individuals of each ecotype into shade cloth at each garden site to measure individual plant responses to precipitation along the Great Plains climatic gradient. An additional 10 blocks (already in place) will also receive a rain out treatment at all sites next spring/summer 2010.
Impacts
Greenhouse Experiments Characterizing Ecotypes. In Fall 2008 and 2009, seed was collected from three precipitation regions (from eastern IL eastern, KS and western KS,) that encompassed a total of twelve native prairie sites, 4 from each precipitation zone. A subset of these seeds were tested at the KS Seed Crop Improvement Center for germination, viability and % live seed. When grown from seed in the greenhouse under similar well-watered conditions, plants from the 3 ecoregions showed significant differences in morphology and physiology suggesting a genetic basis for these traits. The dwarfed western ecotype showed pronounced adaptation to drought including reduced blade area, lower specific leaf area, thicker leaves, greater root:shoot compared to the Illinois ecotype. In addition, the dwarf appearance of the western KS ecotype is related to an internode length that is one third that of the eastern IL ecotype. Physiologically, the western KS ecotype has a significantly lower chlorophyll fluorescence (Fs) and reduced photosynthetic efficiency compared to the eastern IL ecotype. All of these attributes are consistent with our hypotheses of local ecotypic adaptation to drought.Extra sets of plants of these ecotypes have been phenotyped, and used for AFLP marker analysis. Field Experiment-Reciprocal Garden Transplant Studies. In Spring 2009, seed that was collected in 2008 from the 12 populations were used to establish the reciprocal garden experiment as proposed in the USDA grant. Reciprocal common gardens were established in each of the precipitation zones to measure genotype by environment interaction and to detect evidence of adaptive differentiation. Plant establishment data in the reciprocal gardens from the first season already suggest local ecotypic adaptation. At the western sites, in Hays and Colby, KS, the Hays ecotype was represented by a significantly greater numbers of plants than the other ecotypes, which suggests adaptation of the Hays ecotype to areas of low precipitation. Genomic Tool Development. We will begin to develop a new set of genomic tools for big bluestem, instead of relying on the maize cross species hybridization and associated difficulties. We will also use the same group of plants to collect mRNA and construct enriched cDNA libraries to be analyzed for gene expression using the 454 Genome Sequencer. As we gather more information about selected big bluestem genes, eventually we plan to develop a Nimblegen big bluestem custom array for expression studies. We are now doing a 454 sequencing run on cDNA libraries from experimentally droughted big bluestem to detect differential response of the ecotypes.
Publications
- No publications reported this period
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