Source: PENNSYLVANIA STATE UNIVERSITY submitted to
OZONE MONITORING IN NON-URBAN REGIONS OF PENNSYLVANIA AND EFFECTS OF OZONE ON VEGETATION
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
MCINTIRE-STENNIS
Reporting Frequency
Annual
Accession No.
1002837
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
May 1, 2014
Project End Date
Apr 30, 2019
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Project Director
Davis, DO.
Recipient Organization
PENNSYLVANIA STATE UNIVERSITY
208 MUELLER LABORATORY
UNIVERSITY PARK,PA 16802
Performing Department
Plant Pathology & Environmental Microbiology
Non Technical Summary
The US EPA reviews current research and air monitoring data every 5 years for inclusion within the Air Quality Criteria Documents, in order to establish secondary National Ambient Air Quality Standards (NAAQS). Ozone datasets obtained from the proposed monitoring efforts in rural parts of Pennsylvania, as well as the biological effects research, can be used as input when establishing the levels of NAAQS for ozone, which can then be incorporated into federal datasets for other uses. The ambient air data obtained from these non-urban areas will also be used to increase public awareness of the importance of ozone in rural areas, through outreach and educational activities related directly to the proposed monitoring effort. The project will have three main objectives: 1) to monitor ambient ground-level ozone in rural agricultural or forested areas (non-urban areas), 2) to study the biological effects of ozone on vegetation, and 3) through outreach activities, to inform the public as to the adverse effects of ground-level ozone on agriculture and forestry.
Animal Health Component
50%
Research Effort Categories
Basic
25%
Applied
50%
Developmental
25%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1410410106020%
1410699106015%
1412499106015%
2140410106010%
2140699106020%
2142499106020%
Knowledge Area
214 - Vertebrates, Mollusks, and Other Pests Affecting Plants; 141 - Air Resource Protection and Management;

Subject Of Investigation
0410 - Air; 0699 - Trees, forests, and forest products, general; 2499 - Plant research, general;

Field Of Science
1060 - Biology (whole systems);
Goals / Objectives
1. Monitor concentrations of ambient ground-level ozone in rural (non-urban) parts of Pennsylvania. 2. Evaluate effects of ozone on selected plant species. 3. Develop additional bioindicators that can be used to evaluate effects of ambient ozone on forest and agricultural vegetation. 4. Conduct outreach activities regarding ozone effects on vegetation.
Project Methods
1. Continuously monitor ambient ozone concentrations using API Model 400A Ozone Analyzers (EPA-approved equivalent method analyzers) for 12 months/year at four Pennsylvania monitoring sites: Gleason (Tioga Co.), Elliott State Park (Clearfield Co.), State College (Centre Co.), and the Towanda area (Bradford Co.). We will use EPA guidelines for siting and maintaining quality assurance, and provide BAQ with real-time access to the ambient ozone data collected at each site. 2. Expose suspected ozone-sensitive plants (agricultural and forest species) to realistic, ambient levels of ozone as determined by our monitoring efforts. The biological effects research will be conducted within Continuously Stirred Tank Reactor (CSTR) chambers located within the Plant Pathology charcoal-filtered-air greenhouse at the Forestry Research Laboratory on Penn State campus. Results from these exposures will reveal ozone-sensitive plant species, and will also illustrate the threshold levels of ambient ozone needed to cause adverse effects to agricultural crops and native forests within non-urban areas of Pennsylvania. 3. Develop biomonitors that can be used by the private, public, and government agencies (e.g., National Park Service, USDA, USDA Forest Service, US Fish & Wildlife Service) to assess ozone injury on sensitive plant species found in sections of the US where ozone stress to plant life has not been evaluated. We will use the CSTR chambers to expose the plants to realistic ozone concentrations, in order to develop realistic, ozone-sensitive biomonitors. 4. Hold workshops and classes, and provide educational tours at the Air Quality Learning and Demonstration Center within the Arboretum at Penn State, for hosting local organizations and educational classes (from grade school through post-secondary). We will incorporate discussions concerning ozone sources, transport, and impacts on plant growth and development. Ozone impacts will be determined by our observations of plants in open-top exposure chambers within the Air Quality Center, as well as from ozone-sensitive bioindicators planted at the Center. We will also develop documentation (e.g., digital images and descriptive text) of the visual injury response of the most promising biomonitoring plant species for training public, private, and government field crews in diagnosing ozone symptoms on vegetation during ozone-injury surveys.

Progress 05/01/14 to 04/30/19

Outputs
Target Audience:Target audiences include educators, citizen groups, ecology groups, commercial crop producers, the scientific community, the U.S. Environmental Protection Agency, and the Pennsylvania Department of Environmental Protection - Bureau of Air Quality. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project provided training and professional development for numerous Penn State undergraduate and graduate Penn State students. For example, graduate students from the Davis and Decoteau ozone labs who benefited from this project included Lauren Seiler (M.S. awarded), Abigail Myers (M.S. awarded), Kirsten Lloyd (PhD awarded), and Melissa Mercado (M.S. currently enrolled). In addition, the Carlson hardwood genomics program collaborated with faculty member from the University of West Alabama, a predominantly minority-serving undergraduate institution, to provide research experience to undergraduate students. Faculty and a team of undergraduate students from the University of West Alabama visited Penn State University for 2 months each summer. The students were introduced to scientific research focused on ozone exposures, and they received hands-on training in a greenhouse housing CSTR ozone exposure chambers, and in a molecular laboratory. How have the results been disseminated to communities of interest?The Air Quality Learning and Demonstration Center located at Penn State provided hands on learning experiences for classes at Penn State and the general public around Centre County in Pennsylvania. The results of the hardwood genomics project are available online at http://www.hardwoodgenomics.org. The research experience of interns is revealed in short videos (http://www.hardwoodgenomics.org/outreach, https://www.youtube.com/watch?v=2eS8OitPCiQ&feature=youtu.be). Further dissemination has been through publications in scientific journals and presentations at research conferences. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? 1. Monitoring ambient air pollutants Using EPA-approved analyzers, we monitored ambient air quality year-round at the following Pennsylvania rural locations: Gleason (Tioga Co.), Elliott State Park (Clearfield Co.), State College (Centre Co.), and Towanda Area (Bradford Co.). At each site, we monitored ozone, sulfur dioxide, nitrogen oxide, and particulate matter. Equipment was updated as needed with new models, plus personnel were trained on operation and calibration procedures. Monitoring equipment was maintained within allowable drift tolerances for daily span and precision checks, per USEPA requirements within the Federal Clean Air Acts, with all with calibrations completed as directed by Pennsylvania Department of Environmental Protection (PADEP) - Bureau of Air Quality. Flow verifications were conducted monthly on Federal Reference Method (FRM) monitors and Beta Attenuated Monitors (BAM) for particulate matter, with 6-month calibrations completed routinely. Weekly site status reports of all site work and maintenance were provided to PADEP for data validation, quality control, and quality assurance. 2. Evaluating Effects of Ozone on Plants Milkweeds Eleven milkweed species were exposed to ozone within continuous stirred tank reactor (CSTR) chambers in a greenhouse to determine species sensitivity. Common, oval-leaf, prairie, showy, spider, swamp, tall, and tropical milkweed were sensitive to ozone. Butterfly, green, and whorled milkweed were tolerant to ozone. Foliar stipple on common milkweed increased with ozone concentration and duration of exposure. Snap Beans The effect of nighttime ozone exposure, alone and in combination with daytime ozone treatment, was evaluated on yield of ozone-resistant (R123) and ozone-sensitive (S156) snap bean genotypes. Experiments, with exposure durations ranging from 14 to 21 days, were conducted in continuous stirred tank reactor (CSTR) ozone exposure chambers in a greenhouse. The effects of day-only and day + night exposure timings were compared. In a preliminary trial we also tested the effect of nighttime-only ozone exposure. Nighttime ozone exposure alone did not cause foliar injury and had no effect on the yield of either genotype. In combination with daytime ozone exposure, nighttime ozone concentrations did not impact yields or show a consistent effect on nocturnal stomatal conductance. When data were pooled across the day and day + night exposures times, mean daytime ozone levels caused foliar injury and significant yield decreases. Genotype R123 produced significantly greater yield mass than did S156 when exposed to elevated ozone. Nighttime conductance measurements suggest that S156 and R123 have inherently different stomatal conductance rates and that cumulative ozone exposure can influence conductance rates in both genotypes. Overall, results indicate that nighttime ozone exposure has the potential to cause injury to sensitive plants. Therefore, data on nocturnal ozone effects are important data to be input into EPA's next National Ambient Air Quality Standard (NAAQS) for ozone. Trees The Carlson group completed work on the production of transcriptomes (RNA sequence data) for control and ozone-treated seedlings of black walnut, green ash, tulip-poplar, sugar maple, sweet gum, honey locust, and black gum. Results revealed that expression of stress-response, senescence and programmed cell death genes were generally up-regulated, while genes involved in the photosynthetic apparatus were highly down regulated in all species. The initial gene network analyses indicated that ozone stress triggers similar pathways in hardwoods to those triggered in plants by other abiotic stressors. However substantial differences were observed between species that were more tolerant of ozone stress versus those more sensitive to ozone. This suggests that a more detailed study of the regulation of gene expression based on our data may provide candidate genes for improving resistance to oxidative stress in trees. 3. Develop additional ozone-sensitive bioindicators Using our CSTR chambers in a greenhouse, we exposed 11 milkweed species to evaluate their potential use as bioindicators to detect phytotoxic levels of ambient ozone. Common, oval-leaf, prairie, showy, spider, swamp, tall, and tropical milkweed all developed classic ozone-induced dark stipple on the adaxial surface of older leaves and may serve as suitable ozone-sensitive bioindicators to detect phytotoxic levels of ambient ozone, but further testing must be conducted. Also, tropical milkweed also exhibited considerable accelerated leaf senescence (premature defoliation), limiting its ability to serve as an ozone bioindicator. Butterfly, green, and whorled milkweed species were tolerant to ozone, and are not suitable bioindicators of ambient, phytotoxic concentrations of ozone. 4. Conduct outreach activities regarding ozone effects on vegetation. During 2014 - 2018, approximately 600 individuals attended course lectures or public presentations on air pollution effects on terrestrial plants at the Air Quality Learning and Demonstration Center (Learning Center) on Penn State campus. Summaries of the outreach efforts of the Learning Center were presented at the Air Pollution Global Change Symposium (2015) in Monterey, CA and the EPA NOW Conference (2018) in Austin, TX and culminated in the publication "Teaching air pollution effects on plants" (Agri Res & Tech. 11:1-2). In addition, a 3 credit (ERM/PPEM 430) Air Pollution Effects on Terrestrial Ecosystems course was taught to approximately 30 - 35 students each Spring semester at Penn State that included research summaries and tours of the Learning Center.

Publications

  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Davis, D.D. and D.R. Decoteau. 2018. A review: Effect of ozone on milkweeds (Asclepias spp.) in USA and potential implications for Monarch butterflies. Journal of Agriculture and Environmental Sciences 7:156-1172
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Decoteau, D.R. R.P. Marini and D.D. Davis. 2019. Influence of ambient ozone on grapevine cultivars 'Chambourcin' and 'Vidal' in Pennsylvania, USA. Journal of Plant Science & Research 6:185, 5 pp.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2019 Citation: Lloyd, K.L., D. D. Davis, R.P. Marini, D.R. Decoteau, A. Huff and W.H. Brune. 2019. Use of dried compressed air to generate ozone in vegetation exposure studies: quantification of trace nitrogen oxidants formed during corona discharge. Journal of Plant Science & Research 6:pp (in press).
  • Type: Journal Articles Status: Accepted Year Published: 2019 Citation: Seiler, L.K., D.R. Decoteau, R. Marini, and D.D. Davis. 2019. Staghorn sumac (Rhus typhina): a new bioindicator to detect phytotoxic levels of ambient ozone in eastern U.S. Northeastern Naturalist (accepted).
  • Type: Journal Articles Status: Accepted Year Published: 2019 Citation: Lloyd, K.L., D. D. Davis, R.P. Marini, and D.R. Decoteau. 2019. Response of sensitive and resistant snap bean genotypes to nighttime ozone concentration. Journal American Society of Horticultural Science (accepted pending revisions).
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Myers A.C., D.R. Decoteau, R. Marini, and D.D. Davis. 2018. Sensitivity of eleven milkweed (Asclepias) species to ozone. Northeastern Naturalist 25:265-276.


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Target audiences include state forestry personnel; extension personnel; high school, college, and, university educators; commercial crop producers; the scientific community; the U.S. Environmental Protection Agency; the USDA Forest Service; and the Pennsylvania Department of Environmental Protection - Bureau of Air Quality. Efforts were made to deliver science-based knowledge by making oral presentations at formal and non-formal educational and scientific conferences and meetings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Dissemination has been through publications in scientific journals and presentations at research conferences. What do you plan to do during the next reporting period to accomplish the goals?PA DEP furnished us several electronic air pollution monitors at no expense. We will use them to monitor PM2.5 remote northern PA. We will continue to evaluate responses of sensitive and resistant snap bean genotypes and additional milkweeds to ozone. We will also continue to evaluate identify additional milkweed species that can serve as bioindicators to detect phytotoxic levels of ambient ozone in the field.

Impacts
What was accomplished under these goals? We monitored ambient air quality in 2018 at the following rural locations (PA DEP Bureau of Air Quality monitors in urban areas of PA): Gleason (Tioga Co.), Elliott State Park (Clearfield Co.), State College (Centre Co.), and Towanda area (Bradford Co.). We monitored ozone, sulfur dioxide, nitrogen oxide, and particulate matter at these locations. Equipment was updated with new models, plus personnel were trained on operation and calibration procedures. Monitoring equipment was kept within allowable drift tolerances for daily span and precision checks, per USEPA requirements by the federal Clean Air Acts, with all with calibrations completed as directed by Pennsylvania Department of Environmental Protection (PADEP), Bureau of Air Quality. Flow verifications were conducted monthly on Federal Reference Method (FRM) monitors and Beta Attenuated Monitors (BAM) for particulate matter, with 6-month calibrations completed. Weekly site status reports of all site work and maintenance were provided to the state (PADEP) for data validation, quality control, and quality assurance purposes. The effect of nighttime ozone exposure, alone and in combination with daytime ozone treatment, was evaluated on the yield of an ozone-resistant (R123) and an ozone- sensitive (S156) snap bean genotype. Three experiments, with exposure durations ranging in length from 14 to 21 days, were conducted in continuous stirred tank reactor (CSTR) ozone exposure chambers in a greenhouse. The effects of day-only and day + night exposure timings were compared. In a preliminary trial we also tested the effect of nighttime-only ozone exposure. Nighttime ozone exposure alone did not cause foliar injury and had no effect on the yield of either genotype. In combination with daytime ozone exposure, nighttime ozone concentrations did not impact yields or show a consistent effect on nocturnal stomatal conductance. When data were pooled across the day and day + night exposures times, mean daytime ozone levels caused foliar injury and significant yield decreases in all three trials. In all three trials, genotype R123 produced significantly greater yield mass than did S156 when exposed to elevated ozone. Nighttime conductance measurements suggest that S156 and R123 have inherently different stomatal conductance rates and that cumulative ozone exposure can influence conductance rates in both genotypes Using our CSTR chambers in a greenhouse, we exposed 11 milkweed species to determine species sensitivity to ozone, and their potential use as bioindicators to detect phytotoxic levels of ambient ozone. Common, oval-leaf, prairie, showy, spider, swamp, tall, and tropical milkweed all developed classic ozone-induced dark stipple on the adaxial surface of older leaves and may serve as suitable ozone-sensitive bioindicators to detect ambient ozone. However, tropical milkweed also exhibited considerable accelerated leaf senescence (premature defoliation), limiting its ability to serve as a bioindicator. Butterfly, green, and whorled milkweed species were tolerant to ozone, and therefore not suitable to serve as bioindicators. The Carlson group and colleagues continued to test and publish on RNA sequence data from red alder and white alder seedlings during ozone-stress treatments from which transcriptome were assembled and hundreds of microsatellite DNA markers identified. A publication on the results is in preparation and the data is available in the database "www.hardwoodgenomics.org."

Publications

  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Lloyd, K., D.R. Decoteau, R. Marini, and D.D. Davis. 2018. Effects of nightime ozone treatment at ambient concentrations on sensitive and resistant snap bean genotypes. J. Amer. Soc. Hort. Sci. 143:22-33.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Decoteau, D.R., R.P. Marini, and D.D. Davis. 2018. Ozone sensitivity of green ash selections from Midwestern USA. J. Agric. Environ. Sci. on-line
  • Type: Journal Articles Status: Under Review Year Published: 2019 Citation: Lianna J. Johnson, Kathy Haiby, Carlos Gantz, Di Wu, Teodora Best, T. Casey Weathers, Alex Stanish, Brian Stanton, and John E. Carlson, "Development of species specific nuclear SSR markers used to evaluate a controlled hybridization program between red (Alnus rubra) and white (Alnus rhombifolia) alders." (Includes RNA sequence data from ozone-stressed alder seedlings).


Progress 10/01/16 to 09/30/17

Outputs
Target Audience:Target audiences include educators, commercial crop producers, the scientific community, the U.S. Environmental Protection Agency, and the Pennsylvania Department of Environmental Protection. Efforts were made to deliver science-based knowledge by making oral presentations at formal and non-formal educational and scientific conferences and meetings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Through publications in scientific journals and at research conferences. How have the results been disseminated to communities of interest?Through publications in scientific journals and at research conferences. What do you plan to do during the next reporting period to accomplish the goals?We have obtained additional electronic air pollution monitors from PA DEP, at no expense. We plan to improve our ozone and PM2.5 monitoring field efforts in northern PA using this additional equipment. Within the controlled environment chambers, further experiments are planned to explain the observed responses of sensitive and resistant snap bean genotypes to ozone exposure. Experiments will be focused on variation in ozone uptake and sensitivity over time. Additional milkweed species will be exposed to ozone within the greenhouse, in order to determine additional milkweed species' sensitivity to ozone and potential use as additional bioindicators to detect phytotoxic levels of ambient ozone in the field.

Impacts
What was accomplished under these goals? Ambient air quality was measured throughout 2017 at the following locations: Gleason (Tioga Co.), Elliott State Park (Clearfield Co.), State College (Centre Co.), and Towanda area (Bradford Co.). At all monitoring sites, air monitoring equipment was updated for newest models available, plus training on operation and calibration procedures. All air monitoring equipment for ground level pollutants (ozone, sulfur dioxide, nitrogen oxide, and particulate matter) was kept within allowable drift tolerances for daily span and precision checks, per EPA requirements for the federal Clean Air Acts, with all with calibrations completed as directed by Pennsylvania Department of Environmental Protection (PADEP). Flow verifications were conducted monthly on Federal Reference Method (FRM) monitors and Beta Attenuated Monitors (BAM) for particulate matter with 6-month calibrations completed. Weekly site status reports of all site work and maintenance were provided to PADEP for data validation, quality control, and quality assurance purposes. Nighttime ozone exposure trials were continued in the greenhouse to determine the potential for injury to sensitive plants using United States Department of Agriculture snap bean (Phaseolus vulgaris) biomonitoring system. In order to determine the threshold for yield reductions due to nighttime ozone exposure, another trial was conducted in spring 2017 using the USDA biomonitoring system. Over 20 days of treatment, all plants were exposed to daytime mean ozone concentrations of 63 ppb in combination with 11 nighttime treatments ranging from means of 16 to 260 ppb. Results generally supported observations from the summer 2016 trial, indicating greater yield losses with increasing nighttime ozone and a more severe impact on the sensitive genotype. However, nighttime ozone treatment caused relatively higher yield decreases in combination with higher daytime concentrations compared to summer 2016 (mean ozone = 45 ppb). During summer 2017, two short term trials indicated that ozone uptake via stomatal openings on the upper side (adaxial) of snap bean leaves may contribute to ozone injury during both the day and night. Many studies in the published literature have ignored adaxial stomatal openings as a potential route of ozone exposure and injury. Two additional long-term, replicated trials were completed in fall 2017, but results are still being analyzed. Data on nocturnal ozone effects are important for refining EPA's NAAQS for ozone. Eleven milkweed species were exposed to ozone within continuous stirred tank reactor (CSTR) chambers in a greenhouse to determine species sensitivity and potential use as bioindicators to detect phytotoxic levels of ambient ozone. Common Milkweed, Oval-leaf Milkweed, Prairie Milkweed, Showy Milkweed, Spider Milkweed, Swamp Milkweed, Tall Milkweed, and Tropical Milkweed developed typical ozone-induced dark stipple on the adaxial surface of older leaves. Tropical Milkweed also exhibited significant premature defoliation (accelerated leaf senescence). Butterfly Milkweed, Green Milkweed, and Whorled Milkweed were tolerant to ozone. Foliar stipple on Common Milkweed increased with ozone concentration and time. In addition to Common Milkweed, a bioindicator commonly used to detect phytotoxic levels of ozone, the other 8 ozone-sensitive milkweed species should be evaluated further as potential bioindicators. The Carlson group and colleagues continued to test and publish on microsatellite DNA markers derived from RNA sequence data from ozone-stressed hardwood tree seedlings obtained the NSF-supported Hardwood genomics resources project. Two studies were on DNA markers in honeylocust (Gleditsia triacanthos) and the third study on sugar maple (Acer saccharum Marsh.). In addition, RNA sequence data was produced from red alder and white alder seedlings during ozone-stress treatments from which transcriptome were assembled and hundreds of microsatellite DNA markers identified. A publication on the results is in preparation and the data is available in the database "www.hardwoodgenomics.org." This research was supported by The Schatz Center for Tree Molecular Genetics at Penn State and by grant # TRPGR IOS-1025974 from the National Science Foundation's Plant Genome Research Program, completed in July, 2015.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Myers A.C., D.R. Decoteau, R. Marini, and D.D. Davis. 2017. Sensitivity of eleven milkweed (Asclepias) species to ozone. Northeastern Naturalist XX:pp-pp. (accepted)
  • Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Lloyd, K., D.R. Decoteau, R. Marini, and D.D. Davis. 2017. Night ozone treatment does not cause additive injury in sensitive snap bean genotypes. Journal of the American Society for Horticultural Science X:pp-pp. (accepted).
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Decoteau D.R., K. Lloyd, and D.D. Davis. 2017. Teaching Air Pollution Effects on Plants. Agri. Res. & Tech.: Open Access J.: 11(5): 555825.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Harmon M, Lane T, Staton M, Coggeshall MV, Best T, Chen CC, Liang H, Zembower N, Drautz-Moses DI, Hwee YZ, Schuster SC, Schlarbaum SE, Carlson JE, and Gailing O. 2017. Development of novel genic microsatellite markers from transcriptome sequencing in sugar maple (Acer saccharum Marsh.). BMC Research Notes, 10(1), p.369.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Wu, Y., Zhang, R., Staton, M., Schlarbaum, S., Coggeshall, M., Romero-Severson, J., Carlson, J., Liang, H., Xu, Y., Drautz-Moses, D., Schuster, S., Gailing, O. 2017. Development of genic and genomic microsatellites in Gleditsia triacanthos L. (Fabaceae) using Illumina sequencing. Annals of Forest Research 60(2), Online First: June 29, 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Gailing O, Staton ME, Lane T, Schlarbaum SE, Nipper R, Owusu SA, and Carlson JE. 2017. Construction of a Framework Genetic Linkage Map in Gleditsia triacanthos L. Plant Molecular Biology Reporter, Volume 35(2): 177187, (doi:10.1007/s11105-016-1012-0).


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Target audiences include educators, commercial crop producers, the scientific community, the U.S. Environmental Protection Agency, and the Pennsylvania Department of Environmental Protection. Efforts were made to deliver science-based knowledge by making oral presentations at formal and non-formal educational and scientific conferences and meetings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?For the previous five years (2011 to 2015) the Carlson lab hosted undergraduate research interns each summer from the University of West Alabama, a predominantly minority-serving undergraduate institution. A total of 12 undergraduate students from the University of West Alabama, who were introduced to and directly involved in, scientific research focused on ozone stress biology, and featuring gene expression and bioinformatics studies. Of the 12 students, 9 went on to graduate school or professional school for post-baccalaureate training in life sciences. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? At four different rural monitoring sites in Pennsylvania, ambient air quality was measured throughout 2016: Gleason (Tioga Co.), Elliott State Park (Clearfield Co.), State College (Centre Co.), and Towanda area (Bradford Co.). All air monitoring equipment for ground level pollutants (ozone, sulfur dioxide, nitrogen oxide, and particulate matter) was kept within allowable drift tolerances for daily span and precision checks, per EPA requirements for the federal Clean Air Acts, with all with calibrations completed as directed by Pennsylvania Department of Environmental Protection (PADEP). Flow verifications were conducted monthly on Federal Reference Method (FRM) monitors and Beta Attenuated Monitors (BAM) for particulate matter with 6-month calibrations completed. Weekly site status reports of all site work and maintenance were provided to PADEP for data validation, quality control, and quality assurance purposes. Nighttime ozone exposure trials were continued in the greenhouse to determine the potential for injury to sensitive plants using United States Department of Agriculture snap bean (Phaseolus vulgaris) biomonitoring system. The spring 2015 trial, where snap beans were treated with ozone levels of 45 and 75 ppb at two timings (day and full-day exposures), was replicated during fall 2015. Similar to previous results, the timing of exposure had no effect on pod mass at the ozone levels tested over a 21-day period. However, when data were pooled across day and full-day exposures, ozone exposure at 75 ppb significantly reduced the yield of the sensitive genotype, relative to the low (45 ppb) and ambient treatments, in all three trials to date. Nocturnal stomatal conductance measurements continued to show higher gas exchange in the sensitive than resistant bean genotype, potentially leading to higher nighttime ozone exposures. In order to determine the threshold for yield reductions due to nighttime ozone exposure, another trial was conducted in summer 2016 using the USDA biomonitoring system. During the 15-day trial, all plants were exposed to daytime mean ozone concentrations of 45 ppb in combination with 11 nighttime treatments ranging from means of 8 to 265 ppb. Results showed that the yield of both genotypes was significantly affected by the ozone exposures, but ozone caused a greater decrease in the pod mass of the sensitive genotype than the resistant genotype (about 2.2 times more per 1 ppb ozone). Nighttime exposure at 172 ppb reduced pod mass of the sensitive genotype by 50% with a negligible impact on the resistant genotype. Data on nocturnal ozone effects are important for refining EPA's NAAQS for ozone. One plant species commonly used as an ozone-sensitive bioindicator is the ozone-sensitive common milkweed (Asclepias syriaca). We evaluated and compared the ozone-sensitivity of five different milkweed species with the ozone-sensitive common milkweed to determine if other milkweed species could be used as bioindicators of phytotoxic levels of ambient ozone. One of our studies indicated that other milkweed species, except A. curassavica, were comparable in ozone-sensitivity to A. syriaca. A second study revealed that A. syriaca was a viable bioindicator to detect very low levels of ambient phytotoxic ozone. The Carlson group completed work on the production of transcriptomes (RNA sequence data) for control and ozone-treated seedlings of black walnut (Juglans nigra), green ash (Fraxinus pennsylvanica), tulip poplar (Liriodendron tulipifera), sugar maple (Acer saccharum), sweet gum (Liquidambar styraciflua), honey locust (Gleditsia triacanthos), and blackgum (Nyssa sylvatica). Preliminary analysis and comparison of the data revealed that expression of stress-response, senescence and programmed cell death genes were generally up-regulated, while genes involved in the photosynthetic apparatus were highly down regulated in all species. The initial gene network analyses indicated that ozone stress triggers similar pathways in hardwoods to those triggered in plants by other abiotic stressors. However substantial differences were observed between species that were more tolerant of ozone stress versus those more sensitive to ozone. This suggests that a more detailed study of the regulation of gene expression based on our data may provide candidate genes for improving resistance to oxidative stress in trees. The initial transcriptome results were reported in three peer-review publications in 2016, and the data reside in a curated project database- "www.hardwoodgenomics. org." This research was supported by The Schatz Center for Tree Molecular Genetics at Penn State and by grant # TRPGR IOS-1025974 from the National Science Foundation's Plant Genome Research Program, completed in July, 2015.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Carroll RA, Jones C, Best T, Shumaker K, Carlson J. 2016. Gene expression in hardwood trees species exposed to ozone. Journal of Undergraduate Research and Scholarly Excellence, Volume VII: 32-35.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Odom T, Williams KN, Best T, Zembower N, Shumaker K, Carlson J. 2016. An analysis of gene expression induced by elevated atmospheric ozone in hardwood trees native to Eastern North America. Journal of Undergraduate Research and Scholarly Excellence, Volume VII: 36-40.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Lane T, Best T, Zembower N, Davitt J, Henry N, Xu Y, Koch J, Liang H, McGraw J, Schuster S, Shim D, Coggeshall M, Carlson JE, Staton ME. 2016. The green ash transcriptome and identification of genes responding to abiotic and biotic stresses. BMC Genomics 17:702, 16 pages.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Target audiences include educators, commercial crop producers, the scientific community, The Environmental Protection Agency, and the Pennsylvania Department of Environmental Protection. Efforts were made to deliver science-based knowledge by making oral presentations at formal and non-formal educational and scientific conferences and meetings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?For the past several years (2011 to 2015), the Carlson hardwood genomics program collaborated with minority faculty member Ketia Shumaker from the University of West Alabama, a predominantly minority-serving undergraduate institution, to provide research experience to undergraduate students. Dr. Shumaker and a team of 12 undergraduate students from the University of West Alabama visited Penn State University for 2 months each summer. The students were introduced to scientific research focused on ozone exposures, and they received hands-on training in a greenhouse housing CSTR ozone exposure chambers, and in a molecular laboratory. The genetics research was supported by The Schatz Center for Tree Molecular Genetics at Penn State, by a grant from the USDA NIFA Plant Genome Program (#2008-35300-19234), and by grant # TRPGR IOS-1025974 from the National Science Foundation's Plant Genome Research Program. How have the results been disseminated to communities of interest?The results of the hardwood genomics project are available online at http://www.hardwoodgenomics.org. The research experience of interns is revealed in short videos (http://www.hardwoodgenomics.org/outreach, https://www.youtube.com/watch?v=2eS8OitPCiQ&feature=youtu.be). The Air Quality Learning and Demonstration Center provided hands on learning experiences for classes at Penn State and the general public around Centre County in Pennsylvania. During 2014-15 approximately 400 individuals attended course lectures or public presentations on air pollution effects on terrestrial plants at the Learning Center. What do you plan to do during the next reporting period to accomplish the goals?We have submitted two manuscripts for publication and have prepared a third.

Impacts
What was accomplished under these goals? Ambient air quality was measured year-round at four rural monitoring sites in Pennsylvania: Gleason (Tioga Co.), Elliott State Park (Clearfield Co.), State College (Centre Co.), and Towanda area (Bradford Co.). As per EPA requirements for the federal Clean Air Acts, all air monitoring equipment for ground level pollutants (ozone, sulfur dioxide, nitrogen oxide, and particulate matter) was kept within allowable drift tolerances for daily span and precision checks, with calibrations completed as directed by Pennsylvania Department of Environmental Protection (PADEP). Monthly flow verifications were conducted on Federal Reference Method (FRM) monitors and Beta Attenuated Monitors (BAM) for particulate matter with 6-month calibrations completed. Weekly site status reports of all site work and maintenance were provided to PADEP for data validation and quality control and quality assurance purposes. Nighttime ozone exposure was assessed during two greenhouse trials using the United States Department of Agriculture snap bean (Phaseolus vulgaris) biomonitoring system, which uses both a sensitive and resistant genotype. In fall 2014, snap beans were fumigated in continuously-stirred reactor chambers at 30 and 60 ppb ozone and at three different timings: day (8:00-19:00), night (20:00-7:00), or full-day (day + night). After eighteen days of fumigation, results showed that nighttime only fumigation had no effect on seed yield in either genotype. Although none of the treatments had any effect on R123 seed yield, S156 plants exposed to day + night ozone tended to have lower seed mass and seed number than those exposed during the day only. In spring 2015, snap beans were treated with ozone levels of 45 and 75 ppb at two timings: day and full-day. Stomatal conductance measurements taken during the fumigation period showed elevated nighttime gas exchange in the sensitive genotype, particularly for the full-day timing. These results indicate that nighttime ozone exposure has the potential to cause injury to sensitive plants. Data on nocturnal ozone effects are important for refining EPA's secondary National Ambient Air Quality Standard (NAAQS) ozone standard. Asclepias syriaca (common milkweed) seeds were germinated, and resultant seedlings exposed to ozone in continuous stir tank reactors in a carbon-filtered-air greenhouse to determine species sensitivity to ozone. The main objective of the 2014 fumigations was to verify if milkweed can indicate the presence of ozone at a concentration of 60 ppb. Using a regression analysis it was concluded that this species is a viable bioindicator of ozone at this concentration. The characterization of molecular basis of ozone-tolerance/-sensitivity in forest tree species continued in 2014-2015. The main scientific goal was to assess the phenotypical, physiological, and molecular effects of ozone stress on various hardwood tree species. The results of this comprehensive study revealed that ozone induces productivity loss on hardwood tree species regardless of the appearance of tolerance at the visual level, and productivity loss in forest trees is induced even at lower ozone concentrations.

Publications


    Progress 05/01/14 to 09/30/14

    Outputs
    Target Audience: Target audiences include educators, commercial crop producers, the scientific community, The Environmental Protection Agency, and the Pennsylvania Department of Environmental Protection. Efforts were made to deliver science-based knowledge by making oral presentations at formal and non-formal educational and scientific conferences and meetings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? John Carlson collaborated with minority faculty member Ketia Shumaker from the University of West Alabama, a predominantly minority-serving undergraduate institution. Dr. Shumaker and a team of 8 undergraduate students from the University of West Alabama visited Penn State University for two months each summer from 2011 to 2013. The students assessed the effects of ozone stress treatments in various hardwood tree species. The students were introduced to all aspects of the research including growing seedlings in the greenhouse, ozone treatments, RNA extraction, RNA sequencing and gene expression analysis. The students received hands-on training in the Schatz Center for Tree Molecular Genetics and CSTR ozone exposure chambers at FRL. The genetics research was supported by The Schatz Center for Tree Molecular Genetics at Penn State, by a grant from the USDA NIFA Plant Genome Program (#2008-35300-19234), and by grant # TRPGR IOS-1025974 from the National Science Foundation’s Plant Genome Research Program. How have the results been disseminated to communities of interest? The Air Quality Learning and Demonstration Center provided hands on learning experiences for classes at Penn State and the general public around Centre County in Pennsylvania. During 2014 approximately 300 individuals attended course lectures or public presentations on air pollution effects on terrestrial plants at the Learning Center. Dr. Decoteau also presented a summary of the outreach efforts of the Learning center at the Air Pollution Global Change Symposium in Monterrey, CA. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

    Impacts
    What was accomplished under these goals? EPA-approved equivalent method analyzers were used to monitor ambient air quality year-round at four Pennsylvania monitoring sites: Gleason (Tioga Co.), Elliott State Park (Clearfield Co.), State College (Centre Co.), and Towanda area (Bradford Co.). During 2014 new FRM R&P 2025i PM2.5 samplers were installed in State College and a Beta Attenuated Monitor was installed in Gleason. Overall operation, data collection, and reporting of air quality and meteorological data including ground level ozone, sulfur dioxide, nitrogen oxide, particulate matter (PM10, PM2.5), wind speed, wind direction, temperature, relative humidity, rainfall, and solar radiation data was relayed to the Pennsylvania Department of Environmental Protection, Bureau of Air Quality. Asclepias syriaca plants were germinated, and grown and fumigated in continuous stir tank reactors in a carbon-filtered greenhouse at 0,30, 60, and 90ppb to determine species sensitivity to ozone. Asclepias syriaca exhibited foliar damage at 30, 60, and 90ppb ozone suggesting that it is a viable bioindicator of ozone in extremely low ozone concentrations. The characterization of molecular basis of O3-tolerance/-sensitivity in forest tree species included three new forest species; Sugar maple (Acer saccharum), Sweet gum (Liquidambar styraciflua) and Honey locust(Gleditsia triacanthos). Greenhouse controlled studies were conducted for 4 weeks/28 days, 8hr/day using ozone exposures known to induce visible ozone injury in forest tree species (80, 125, and 225 ppb). At the end of the treatment, all 3 species expressed ozone induced visible injury. Leaf tissue was collected for RNA extraction and transcriptome sequencing. The transciptome sequencing and data analysis will provide a deeper understanding of molecular processes and responses involved in plant defense.

    Publications