Brief Summary of Minutes of Annual Meeting

Chair David Grantz welcomed everyone to the meeting and thanked host, Dennis Decoteau, for organizing the meeting this year. We were reminded that the current project expires in 2012 and that we should be thinking about approaches for the new proposal. Grantz encouraged the group to think about possible collaboration activities, funding opportunities and the need to increase student participation. Dennis Decoteau welcomed the group and thanked Jim Savage for his help setting up for the meeting. Jim is the primary caretaker of the Demonstration Center and has done a great job maintaining and improving the facility. Margaret Smith, Project Administrator, encouraged the group to continue meeting its multi-state project objectives. Collaborative research is a component of Hatch Fund projects, and 25% of each projects fund must be spent on it. Our annual report should highlight multi-state research activities. She went on to say that our project renewal process should begin now. The current project terminates on September 30, 2012. A request to write a proposal renewal is due to NIMSS by March 2011. The request should emphasize whats next in the projects objectives and how it will take advantage of the multi-state approach. Ray Knighton, our USDA representative, described the four institutes in the newly formed NIFA office, formerly known as CSREES. The institute of Energy, Environment and Climate Change is where he is now assigned. The focus of the funding program has been narrowed compared to previous years but the scale of the projects is much larger. Resources allocated to each project were increased to promote multidisciplinary teams whose research will have impact, in this case, on climate change mitigation and agricultural production problems associated with climate change. Emphasis will be on projects that can develop approaches that will be viable for use in the field in short order. David Grantz conducted the business meeting. A committee was formed, consisting of Art Chappelka, Fitz Booker and Dave Grantz to write the request to renew the project proposal, due for submission by March 2012. Location of the next meeting will either be on Long Island, hosted by Meg McGrath, or at Rutgers University, hosted by Barbara Zilinskas. Art Chappelka succeeded Dave as Chair of the project. The meeting was adjourned at 3:00 pm, July 16, 2010.

Activities of the participating experiments are guided by the Objectives of the Current approved Project Proposal. These activities and achievements follow, organized by NE-1030 Project Objectives, and by approaches. OBJECTIVE 1. Describe the spatial - temporal characteristics of the adverse effects of current ambient O3 levels on crop productivity, including the development of numerical models to establish cause  effect relationships that apportion the ozone contribution. 1a. spatial analysis of ozone impacts on crops. A collaboration between Booker, Burkey, Fiscus and Ainsworth of USDA/ARS, NC State University and the University of Illinois, considered the elevated concentrations of ground-level O3 that are frequently measured over farmland regions in many parts of the world. While numerous experimental studies show that O3 can significantly decrease crop productivity, independent verifications of yield losses at current ambient O3 concentrations in rural locations are sparse. In this study, soybean crop yield data during a 5-year period over the Midwest of the United States were combined with ground and satellite O3 measurements to provide evidence that yield losses on the order of 10% could be estimated through the use of a multiple linear regression model. Yield loss trends based on both conventional ground-based instrumentation and satellite-derived tropospheric O3 measurements were statistically significant and were consistent with results obtained from open-top chamber experiments conducted by ARS researchers in Raleigh, NC and an open-air experimental facility (SoyFACE) in central Illinois, conducted by ARS researchers there. Extrapolation of these findings supports previous studies that estimate the global economic loss to the farming community of more than $10 billion annually. (NC, IL,). 1b. diurnal trends in ozone sensitivity of vegetation. A collaboration of the University of California at Riverside (R. Heath) University of California Kearney Agricultural Center (D. Grantz), and USDA/ARS (K. Burkey) has addressed the factors that determine the sensitivity of extensive vegetation to ozone. In general these are only crudely characterized. In order to support efforts to model extensive regional impacts, it is necessary to parameterize the steps of ozone injury, and to characterize the variability among populations. There are three major steps for plant injury by ozone (O3). These are entrance of O3into the leaf (Flux, F), overcoming by O3 metabolic defenses (antioxidant capacity, A), and the actual attack of the effective dose of O3 (Deffective) on bioreceptors (Injury, I).This can be expressed mathematically. Current approaches model gs (stomatal conductance) and [O3] from meteorological data, the species composition of the ground cover, and air quality monitoring data. However, A is assumed to be constant. Recent use of flux (F) is an improvement over previous use of exposure ([O3]) to estimate injury. We have tested the hypothesis that defense capacity (A) may vary diurnally, and may therefore control the amount of injury caused by a given atmospheric ozone concentration. If so a specific O3 flux (F) will yield a different Deffective and thus a different I, at different times of the day, so that this will need to be considered in modeling of regional O3 impacts. From knowledge of how much O3 enters the leaf (F) and how much injury occurs (I), we can calculate the initial defense capacity of the leaf (A). Exposure must be rapid enough that tissues have insufficient time for induction of additional defense capacity. We have developed the first demonstration that defense capacity varies diurnally, and have explored the mechanism, using Pima cotton, cv. S-6, grown in a greenhouse. Injury was determined from digital photo analysis of necrosis, chlorophyll content (SPAD, Minolta) and summed abaxial and adaxial stomatal conductance (LiCor 1600) 6-7 days after exposure. Total antioxidant capacity, ascorbic acid and dehydroascorbic acid content were determined on non-exposed leaves at different times of day. Injury induced by an (interpolated) O3 dose of 19.8 mol m-2, exhibited a clear diurnal trend, shown by foliar necrosis, chlorophyll content (SPAD) and stomatal conductance, all obtained at 6 days after exposure for a 15 minute pulse. Leaves were most sensitive near 3:00 p.m. in repeated experiments. Antioxidant levels of foliar ascorbic acid and of total foliar antioxidant capacity exhibited a moderate peak near midday, but leaf injury was also greatest at this time. Regression relationships between sensitivity to O3 injury and various measures of antioxidant status were not significant. While the diurnal nature of ozone sensitivity is confirmed, the mechanism remains to be elucidated. These data indicate that parameterization of models of O3 injury to vegetation will require measures of inherent defense capability, for which time of day may be a key determinant. (CA, NC) 1c. Snapbean model system demonstrates ozone impacts on crops. The continuation of this study will strengthen our understanding of the impact of ambient ozone on plants and crop productivity. In NY, McGrath has been assessing impact on plant productivity of ambient ozone occurring on Long Island, where she is stationed, by growing the ozone-sensitive and ozone-tolerant snap bean lines that were developed for use in quantifying ozone impact. Each year there have been three successive field plantings to cover the entire growing period for beans in the area. As they developed, bean pods were harvested repeatedly from some plants when immature and at a size typical for fresh-market consumption. Pods were harvested from the other plants when mature and dry. Plants were examined routinely for ozone injury. Injury and defoliation due mainly to ozone injury were rated. Ozone concentration data were obtained from a monitor maintained at the research site (LIHREC) by the NYC DEC Air Quality Division. Results from 2009 research were analyzed and another experiment was conducted over the past year for this reporting period. In 2009, the ozone-sensitive snap bean line S156 yielded less than the tolerant line R331 when grown under ambient ozone conditions. Total weight and number of bean pods harvested for fresh-market consumption from planting 1 (22 May) plants was 28% and 19% lower, respectively, for S156 compared to R331 (pods were harvested from 17 July through 21 Aug). There was a 55% and 46% reduction in these yield variables, respectively, for planting 2 (22 June) plants (harvested 10 Aug through 9 Sept). Reduction was 33% and 16%, respectively, for planting 3 (16 July) plants (harvested 4 Sept through 6 Oct). These differences were similar to greater than in previous years. Mature yield data has not yet been collected. Exposure to ozone caused acute foliar injury in all three plantings. The visible symptom was bronzing. The sensitive line became more severely affected than the tolerant one. Severely affected leaves eventually died and dropped. For example, injury was first observed on Planting 1 plants on 19 June. Average percentage of leaf tissue with bronzing (determined by estimating the incidence or proportion of leaflets with symptoms and the average severity on affected leaves) was 0.04% and 5.6% for R331 and S156, respectively, on 11 July, which was 6 days before the first pods were ready for harvest. Average percentage of leaf tissue with bronzing had increased to 0.5% and 64% by 27 July and 2.4% and 70% by 1 Aug. (NY) In New Jersey, Zilinskas monitored the effects of ambient ozone on the productivity of two snapbean cultivars R331 (ozone-tolerant) and S156 (ozone-sensitive) over the 2009 growing season. Following a very wet month of June, we planted the two snapbean cultivars in East Brunswick, NJ, on July 1, 2009, two weeks after our usual planting date. We closely adhered to the field design and conditions agreed upon by the four field stations in the US that are collaborating on this project. Throughout the growing season, ambient ozone levels and meteorological data were recorded at each site. At each field station, we made multiple harvests of marketable pods at 49, 56, 63, 69 and 76 days after planting. In the 2009 season, peak pod number and fresh weight (of both cultivars) occurred at the second harvesting date. A statistically significant decrease in total pod fresh weight of marketable snapbeans was observed in S156 relative to R331 in three of the five harvest dates, where the fresh weight of marketable pods of the ozone-sensitive cultivar was between 37% and 73% less than that of the ozone-tolerant cultivar. A final harvest of snapbeans was conducted at a uniform 84 days after planting. At this harvest date in the 2009 growing season, a significant proportion of the pods were immature (without seeds) or green. The number of seeds and the dry weight of seeds and pods from the two cultivars were significantly different, with yield reductions in the sensitive relative to the tolerant cultivar of 26%, 47% and 50%, respectively. There was no significant difference in the pod number of the sensitive cultivar relative to the tolerant cultivar. (NJ) The meteorological and ozone data, coupled with the crop yield data, will be analyzed for the several states where this field experiment has been conducted and incorporated into a numerical model by Dr. S. Krupa (MN) to establish a relationship between ambient ozone exposures and crop responses. (MN) 1d. ozone impacts on plants with C4 photosynthetic systems. A number of food and potential biofuel crops in California utilize the C4 photosynthetic pathway. This is considerable desirable due to the inherent water use efficiency of this mode of carbon acquisition. In the studies of the National Crop Loss Assessment and more recently (maize reduced by 4-8%, relative to soybean of 22%, for example), these types of plants have been considered to be tolerant of ozone. We decided to investigate genotypes of the Saccharum complex which are being considered as sources of biofuel, both through easily fermented sugars from commercial sugarcane clones, but also for lingo-cellulosic feedstocks from high fiber energy canes, which are relatively low in sugar. Existing genotypes were not developed in areas subject to high ozone. A locally grown clone of sugarcane, favored by farmers of southeast asian descent, was the most sensitive. A commercial sugarcane clone from Texas was reduced in biomass production by 30%, the southeast asian clone was reduced by about 55%, while two clones with high percentage of the wild relative, Saccharum spontaneum, were not significantly affected. The most sensitive clone was inhibited in dry matter production by 38% at 12 hour mean ozone of 59 ppm, and by 75% at 117 ppm. This is substantial sensitivity to ozone, relative even to sensitive crops such as Pima cotton. C4 crops are similar to C3 crops in exhibiting a range of ozone sensitivity. It is not warranted to assume that C4 crops will exhibit the high levels of ozone tolerance observed in early studies. (CA) OBJECTIVE 2. Assess the effects of O3 on structure, function and inter-species competition in managed and native plant populations, including alterations in their nutrient quality. Previous work underway on competition between horseweed, purple nutsedge and the crops cotton and tomato has been concluded. Extension efforts are taking this information to growers in Californias Central Valley (CA) 2a. impacts of ozone on growth, competition and nutritional value of forages. A study was initiated in 2009 to determine the effects of tropospheric ozone and various precipitation regimes on a semi-natural grassland characteristic of the Piedmont region of the US (mixture of tall fescue, common bermudagrass, dallisgrass and white clover). Twelve, large (4.8 m ht. × 4.5 m dia.) open-top chambers (modified with rain-exclusion caps) located at the Auburn University Atmospheric Deposition Site were used in this study. A multifactor design with two ozone treatments [nonfiltered (NF, ambient) and 2X × NF] and 3 water regimes (30-yr average, +20% and -20%) were replicated 2 times. Ozone exposures and rain treatments were initiated June 1, 2009. Primary growth and regrowth forage were harvested monthly during the growing season. In addition, a point-sampling technique was used to determine species abundance and diversity. Forage samples were analyzed for concentrations of cell-wall constituents and crude protein. Data are currently being statistically analyzed. Results will provide critical information on structure and functioning of managed grassland ecosystems using projected climate scenarios of elevated ozone and differing amounts of rainfall, with emphasis on interspecific relationships among the various processes examined. Integration of various measures of diversity and productivity and underlying physiological and biochemical responses will enable a more complete characterization and modeling of potential impacts of future climate change scenarios on these plant communities. Non-fumigated forage from our site was harvested on April 21 and May 12, 2008, after which they were exposed to either ambient, non-filtered air (NF) or twice-ambient O3 air (2 × NF) air and harvested on June 9 and July 2. Forages were fabricated into 50-g cubes that were fed to New Zealand White rabbits in a nutrient-utilization/diet-selection experiment beginning in December 2009. Sixteen, 8 wk-old rabbits were initially obtained, from which 10 were used in the experiment and divided into two groups: 1) fed NF forage and 2) fed 2X forage for 10 days. Orts for each rabbit were collected each day before new forage was introduced and pooled for future chemical analysis for all 10 days. The final six days consisted of fecal and urinary output collection for each rabbit. Orts, feces, and urine were collected from steel trays located under each rabbits cage. Samples were analyzed for various nutrient constituents. Preliminary results indicate that digestible dry matter intake was lower for 2X rabbits compared with NF-fed rabbits, primarily as a result of decreased digestibility of cell-wall constituents. The decrease in fiber digestibility cannot be explained on the basis of lignifications solely as originally hypothesized. Further analysis is being conducted investigating the possible role of soluble phenolics.(AL) OBJECTIVE 3. Examine the joint effects of O3 with other growth regulating factors (e.g., CO2, temperature) that are expected to vary with ongoing climate change on crop growth and productivity. 3a. drought x ozone interactions. A five-year (2004  2008) field study by Decoteau on the effects of ambient ozone levels on the growth and production of two wine grape varieties was terminated in 2009. We have determined through ambient ozone exposures over the past four years at our research site located at the Penn State Fruit Research and Extension Center (Biglerville, Adams Co.) that the grape cultivar Charbourcin is sensitive to ozone levels typically experienced during the summer months in Pennsylvania. Ozone injury to Charbourcin grape included adaxial stipple and yellowing and defoliation of the older leaves. The amount of injury observed on the grape foliage varied from year to year and appears to be influenced by other weather conditions. Injury was more significant during the 2004 and 2006 growing seasons. The droughts of 2005 and 2008 resulted in less injury than in other years. The Vidal variety of grape, which is considered tolerant to ozone injury, typically exhibited little to no foliar injury to ambient ozone levels. Each year, berry harvests were made in early October and fruit quality evaluations determined. Berry weight, pH, brix levels and fruit acidity varied from year to year and among treatments and varieties. Grape plants in the open plots tended to have less berry weight and more acid than the other treatments. Further evaluations need to look at the influence of recorded ozone levels and its variability during the season on fruit results. (PA) A model was developed to predict stomatal conductance under well watered and water stressed conditions in soybean using soil moisture and common atmospheric inputs. By including field site measurements of atmospheric O3, the midday leaf influx of O3 may be calculated and related to yield and visible damage effects. (NC) 3b. effect of houseplants on indoor ozone exposures. Three common indoor houseplants, the snake plant, spider plant, and golden pothos, continued to be evaluated for their species effectiveness in reducing ozone concentrations in a simulated indoor environment. Ozone was injected into the continuously stirred tank reactor chambers housed within a greenhouse equipped with a charcoal filtration air supply system chambers and when concentrations reached 200 ppb (+/- 5 ppb), the ozone generating system was turned off and ozone concentrations over time (ozone was monitored every 5 to 6 min in each chamber) were recorded until approximately < 5 ppb was measured in the treatment chamber. On average, ozone depletion time (time from when the ozone generating system was turned off at approximately 200 ppb to < 5 ppb in the chamber) ranged from 38 to 120 min per evaluation. Ozone depletion rates were higher within chambers that contained plants than within control chambers without plants, but there were no plant species differences. New studies are focusing on the impact of total surface area within the chambers on the influence of plant reduction in ozone levels. (PA) 3c. ozone by carbon dioxide interactions. Agricultural soils are thought to be a C sink in a changing global climate because rising CO2 often enhances plant-derived C inputs belowground. However, elevated tropospheric O3 concentrations may offset the CO2-fertilization effect. Unfortunately, limited information is available on long-term belowground responses to elevated CO2 and O3 in agricultural systems. The objective of our five-year experiment was to determine the separate and combined effects of elevated CO2 and O3 on soil respiration, microbial biomass, nutrient availability and soil C levels in a no-till soybean-wheat cropping system using open-top field chambers. Soil respiration during four growing seasons was stimulated by 21% with elevated CO2, but the O3 effect was not statistically significant. Microbial soil respiration and biomass were higher with elevated CO2 at later stages of the study but similar to the control in the O3 treatment. Soil delta 13C values decreased with elevated CO2, showing that elevated CO2 influenced soil C concentrations, but soil C levels were not significantly affected by either elevated CO2 or O3. It is hypothesized that increased residue and root C and N inputs with elevated CO2 accelerated decay rates, as evidenced by increased soil and microbial respiration rates. High N availability due to increased input from an N-fixing legume, along with increased C and adequate supplies of other mineral nutrients and water, may accelerate organic C turnover at elevated CO2, thus constraining potential C sequestration in highly-managed agroecosystems. (NC). 3d. ozone by vapor pressure interactions. Differential atmospheric vapor pressure deficit experiments showed that high vpd substantially reduced seed yields in the S156/R123 bioindicator snapbean pair. At low vpd the addition of O3 reduced yield by 55% (R123) and 72% (S156). High vpd in clean air reduced yields by 30% in both genotypes while the addition of O3 at high vpd had no further significant yield effect in either genotype. However, in O3 air, high vpd had a salutary affect, increasing yields by 43% (R123) and 83% (S156) when compared to the high O3-low vpd treatment. However, these interactions, did not substantially affect the ratio (S156/R123) of responses to O3, thus preserving their usefulness as an O3 bioindicator pair. (NC) 3e. ozone x herbivorous insect interactions. We are also comparing genome-level responses to ozone in two hybrid poplar clones that are known to vary in sensitivity to ozone. In the summer of 2009 the O3-sensitive (NE 388) and O3-tolerant (NE 245) clones were exposed to damaging levels of ozone, with half of the plants also being subjected to gypsy moth caterpillars, Lymantria dispar, a common defoliator of Populus spp. We conducted a full factorial, time course microarray experiment to investigate global gene expression patterns in a spatially and temporally robust manner. Microarray hybridizations were conducted with the recently developed whole genome microarray for Populus from Nimblegen. A first study showed that 73% of the genes that are typically regulated in poplar by herbivory under controlled conditions were not regulated by herbivory when the plants were exposed to ozone, while only 15% of the herbivore-regulated genes were activated independent of ozone. We are now in the process of analyzing the data from the second study. Our results should provide insights into plant adaptations to biotic and abiotic stressors (i.e. O3 and herbivores) by identifying genes and gene regulation networks that are activated in multiply stressed plants. This study should reveal genome-level interactions and will be useful in developing strategies that might provide tolerance to both abiotic and biotic stresses in plants.This project has led to the production of more expressed gene sequences for Black Cherry than are currently available for any other single Rosaceae species. (PA) OBJECTIVE 4. Examine the physiological and molecular basis of O3 toxicity and tolerance in plants. 4a. role of antioxidant defense mechanisms. A collaboration between USDA/ARS in NC and Misericordia University in PA further examined the antioxidant relationship with ozone injury to determine the role of the apoplast in plant defense responses to oxidative stress.. Upon entry into leaves, O3 and related ROS must pass through the leaf extracellular space before reacting with plasma membrane components to initiate plant injury responses. Therefore, O3 and ROS detoxification reactions localized in the leaf cell wall represent a first line of defense against O3 injury. Cellular events caused by O3 exposure include the production of reactive oxygen species (ROS), which can lead to foliar injury and suppressed biomass production. However, plants possess biochemical mechanisms that regulate ROS concentrations, such as peroxidases and superoxide dismutases. In Arabidopsis, stimulation of two cationic peroxidases was observed following exposure to moderate levels of O3. Through the use of protein sequencing, gene expression arrays, quantitative real-time PCR and insertion mutants, identity of the peroxidases was determined, along with confirmation that one gene was transcriptionally regulated in response to O3. Knock-out mutants for the peroxidase genes are being used to investigate possible functional significance of peroxidase activity changes in responses to O3. In a related experiment, Arabidopsis mutants transformed with a superoxide reductase gene, which have shown increased tolerance to heat stress and a ROS-generating herbicide, did not exhibit increased tolerance to O3, suggesting that protection from superoxide in the cytoplasm was relatively unimportant in counteracting O3 toxicity in this experimental system. (NC). Insoluble cell wall material was isolated from leaves of O3-sensitive and tolerant genotypes of soybean and snap bean to assess differences in cell wall chemistry that may relate to observed differences in O3 sensitivity. Snap bean and soybean plants were exposed to charcoal-filtered (CF) air or elevated O3. Leaf intercellular wash fluid (IWF) was recovered and analyzed by reversed-phase HPLC using a column selected for separation of highly polar aromatic compounds.HPLC analysis following alkaline hydrolysis of the cell wall material revealed relatively simple chromatograms consisting of 4 to 6 unknown compounds that varied in quantity depending upon species, genotype, and O3 treatment. Snap bean and soybean plants contained unique profiles of soluble compounds in the leaf apoplast demonstrating significant species differences. In snap bean, no major genotype or ozone treatment effects were observed. In soybean, significant differences in quantities of apoplast constituents were observed when the two genotypes were compared, often with greater amounts present in O3-tolerant Fiskeby than in the O3-sensitive Mandarin Ottawa, regardless of treatment. Several peaks present in both soybean genotypes were reduced by O3 treatment in Mandarin Ottawa but not in Fiskeby. Future work will be directed toward identification of the major apoplast constituents and their role, if any, in O3 tolerance. (NC, PA). Molecular genomic analyses of previously identified ozone tolerant and ozone sensitive families of black cherry and Populus are allowing us to characterize and understand the physiological and molecular basis of ozone toxicity and tolerance in trees. We are currently studying both Populus and Prunus species, taking advantage of previous research that has identified ozone tolerant and sensitive genotypes in these species. For black cherry, a wide range of O3-sensitivity is known, with some genotypes being so sensitive as to serve as ozone indicator plants. We conducted four years of O3 treatments at normal ambient stress levels (80 ppb O3 for 8 hr, 7 day/wk) in three large half-sib (open-pollinated) black cherry families which were known to have low, medium, and high family means for ozone damage. The treatments permitted us to identify the most ozone-sensitive and ozone-tolerant individuals within each family. We prepared two cDNA libraries from RNA isolated from O3-treated leaves of ozone-sensitive and ozone-tolerant seedlings, from which we generated app. 82 million bases of DNA sequence in 2008, and 250 million bases of sequence in 2010. Functionally, 13% of the DNA sequences of the sensitive family are from genes known to be involved in response to biotic and / or abiotic stresses (Figure below). Within these gene sequences we have indentified unique microsatellite DNAs that we are presently using to construct genetic linkage maps for black cherry, with which we plan to map the loci for ozone tolerance. About 1% of genes in black cherry are involved in signal transduction. The full-sib black cherry families that we have selected for mapping will be maintained as reference populations for future research. We have also shown recently that the new DNA markers can be used to identify pollination patterns within the black cherry seed orchard at Penn Nursery. 4b. role of anatomical defense mechanisms. A collaboration between Neufeld of Appalachian State University and many others has characterized impacts of ozone on understory vegetation of Great Smokies National Park. The factors responsible for determining the ozone sensitivity of a plant may range from the physical to the biological. Physical properties of cells (cuticular cell wall and mesophyll cell wall thickness), and leaves (thickness, number of layers of palisade mesophyll cells, cell packing or density, exposed internal cell surface area, the tortuosity of the diffusional pathway, and stomatal densities and sizes) may play a role in determining ozone sensitivity in plants. In this study, leaf anatomical properties were investigated for ozone-sensitive and ozone-insensitive individuals of the ozone bioindicator plant cutleaf coneflower (Rudbeckia laciniata var. digitata) at Purchase Knob in Great Smoky Mountains National Park. Plants growing at Purchase Knob in either a field (full sun) or beneath a forest (shade) were studied. Both young and old leaves on the same plant were studied, but because results were similar for the two age classes in most cases, leaf age is not addressed further in this report. Leaves were sampled in mid-August of 2003, and then preserved in FAA before undergoing dehydration in EtOH. They were fixed in plastic, sectioned on a microtome, stained and viewed using TEM and light microscopy. Image J was used to measure various parameters (described below). There were no statistically significant differences in either adaxial or abaxial stomatal densities between the two groups of plants, or between those growing in sun or shade. Leaves in the sun were thicker than those in the shade, but there were no sensitivity differences. There were also no sensitivity differences for palisade or spongy mesophyll layer thicknesses, but these layers were thinner for leaves grown in shade. There was a significant trend for narrower epidermal wall/cuticle thicknesses in insensitive plants compared to sensitive ones, in both the sun and shade, but not so for either palisade or spongy mesophyll wall thicknesses. Total cell areas appeared to be slightly less in insensitive plants than sensitive ones, mainly due to reduced spongy mesophyll areas, but not palisade mesophyll areas. The amount of airspace did not differ among the sensitivity types. The tortuosity of the diffusional pathway appeared greater in sun plants than shade, and there was a light x sensitivity interaction, mainly due to the fact that tortuosity was higher in ozone-sensitive plants when growing in the sun, but not in any other condition. In the two cases where sensitivity was statistically significant (adaxial epidermal wall/cuticle thickness and spongy mesophyll cell area) it was greater in the sensitive plants, which is contrary to what models would predict for increasing sensitivity. Thicker cell walls would have a greater capacity to store anti-oxidant compounds, and would inhibit the diffusion of ozone the cell membrane, so if this parameter is involved with determining sensitivity, it should be thinner in the sensitive individuals, not the insensitive ones as found. Therefore, we conclude that in cutleaf coneflower, leaf anatomical differences do not contribute to the observed sensitivity differences in this species. Because earlier studies on cutleaf coneflower have shown no differences in gas exchange or anti-oxidant activity in uninjured leaves, we suggest that the basis for sensitivity variation might reside elsewhere, possibly at the molecular level, where ozone may differentially affect gene regulation and/or transcriptional regulation between sensitive and insensitive individuals. (NC) Objective 5. Develop educational tools and conduct advanced training for K-12 public school teachers, college level instructors, and outreach educators regarding the effects of ambient O3 pollution on plants. 5a. community involvement in annual meetings. The NE-1030 Technical Committee conducts outreach in the individual states and communities, and through the annual meeting attempts to forge linkages with local agencies, institutions and personnel. At this years Annual Meeting in State College PA a number of such linkages were developed. A field trip to Accuweather, the headquarters of a weather-forecasting company, was incorporated in the afternoons activities. We wish to express our appreciation to Mark Steinberg, our tour group leader, and Accuweather for a most interesting and informative tour. John Skelly, now retired but a long-time member of the Technical Committee, opened the technical portion of the meeting with a presentation on the history of the Air Quality Learning/Demonstration Center. He emphasized that success of the center was due in part to cooperation between Penn State University and private industry, notably RRI Energy. John said it was great to see how far the center had progressed from his initial efforts in getting it started. John brought Vince Brisini to the meeting, where he was able to meet with the currently active Ozone researchers gathered. Vince Brisini, from RRI Energy Corporation, talked about their participation in establishing the Air Quality Demonstration Center and reviewed the current status of emission control activities at power generation facilities. He stated that emission controls were quite effective now and that attaining further reductions will be difficult. Economic incentives must be available for businesses to comply with stricter regulations. (PA) Holly Salazer, Air Resource Manager, National Park Service, located at Penn State, presented an overview of the park services goals for improving air quality in National Parks. By law, they are mandated to prevent significant deterioration in air quality in National Parks. Specifically, by 2064, the goal of the Regional Haze Regulation is to reduce haze in the parks to pre-impact levels. Hollys office helps coordinate analysis of air quality monitoring data and comments on permit applications for development that may affect air quality in national parks. (PA) Maria Cazorla, a Ph.D. student at Penn State, presented results of her research on a new instrument that measures ozone production rates for use in air quality monitoring in the field. 5b. Public education centers on air quality issues. Decoteau at Penn State maintains the Air Quality Learning and Demonstration center, which continues to provide hands on learning experiences for classes at Penn State and the general public around Centre County in Pennsylvania. During 2009 approximately 250 individuals attended course lectures or public presentations on air pollution effects on terrestrial plants at the Learning Center and in 2010 we hosted the NE 1030 project participants. (PA) Grantz in California maintains field and greenhouse exposure chambers that are shown to visiting school and industry groups and international visitors from commodity outreach programs. (CA) 5c. Expert involvement in local educational and cooperative extension programs. The USDA/ARS group in Raleigh NC (Burkey, Booker, Fiscus) has developed some direct participation programs for NCSU students in the Environmental Technology courses, Plants Soils and Natural Systems (ET202) and Crop Physiology (CS714). Demonstrations were presented using the ozone-sensitive and ozone-resistant snap bean lines to explore effects on photosynthesis, stomatal conductance, biomass production and visible injury due to ozone. The genetic component of differential ozone sensitivity between genotypes was highlighted. Photosynthesis measurements at a range of CO2 concentrations showed that C fixation by the enzyme Rubisco was inhibited by O3 in the sensitive line, but not in the resistant line. This finding demonstrated one biochemical mechanism of O3 effects on plants that likely accounts in part for suppressed biomass accumulation and yield of plants in the field. (NC). Middle school students at Rosman High School, Rosman, NC were provided seeds of ozone-sensitive and ozone-resistant snap bean genotypes and guidance for a 9th grade science fair project on the effects of light and stomatal conductance on plant responses to ozone. (NC). McGrath (NY) shared information on ambient ozone and its impacts during a class on vegetable diseases that is presented annually during the Master Gardener Training Program conducted during spring each year in Suffolk County.(NY) Grantz addressed school groups and industry organizations, and worked with individual commodity groups to explain the importance of air quality improvement. These efforts are beginning to interact with groups seeking to reduce the carbon footprint of energy production, a specific initiative at this time in California. (CA) 5d. Maintenance of a web educational presence. Booker of USDA in NC, the Web Master for the NE-1030 Project, developed and maintained a project web page at (http://www.ncsu.edu/project/usda-ne-1013/index.htm). This web page is frequently updated with current news items, project annual report and news of the project such as minutes of the 2009 annual meeting. (NC).

Booker, F., R. Muntifering, M. McGrath, K. Burkey, D. Decoteau, E. Fiscus, W. Manning, S. Krupa, A. Chappelka, and David Grantz. 2009. The Ozone Component of Global Change: Effects on Agricultural and Horticultural Plant Yield, Product Quality and Interactions With Invasive Species, Journal of Integrative Plant Biology 51: 337-351.

Ditchkoff, S.S., J.S. Lewis, J.C. Lin, R.B. Muntifering, and A.H. Chappelka. 2009. Nutritive quality of highbush blackberry (Rubus argutus) exposed to tropospheric ozone. Rang. Ecol. & Mang. (In press, available online, DOI: 10.2111/08-222.1).

Fishman, J, JK Creilson, PA Parker, EA Ainsworth, GG Vining, J Szarka, FL Booker, X Xu. 2010. An investigation of widespread ozone damage to the soybean crop in the upper Midwest determined from ground-based and satellite measurements. Atmospheric Environment 44:2248-2256.

Gould, Kevin S., Dana A. Dudle and Howard S. Neufeld. 2010. Why some stems are red. Photoprotective roles for anthocyanins in internodes. Journal of Experimental Botany 61:2707-2717.

Grantz, D.A., Shrestha, A., Vu, H. Ozone Impacts on Assimilation and Allocation to Reproductive Sinks in the Vegetatively Propagated C4 Weed, Yellow Nutsedge. Crop Science 50:246-252.

Grantz, DA, Vu, H-B. 2009. O3 Sensitivity in a Potential C4 Bioenergy Crop: Sugarcane in California. Crop Science. 49:18.

Grantz, D.A., Vu, H., Aguilar, C., Rea, M.A. No Interaction Between Methyl Jasmonate and Ozone in Pima Cotton: Growth and allocation respond independently to both. Plant Cell and Environmen. 33, 717728.

Grantz, Da Vu. H-B, Heath, RL, Burkey K. 2010. Temporal Sensitivity Key to Modeling Ozone Impacts on Vegetation. Extended Abstract 2010-EE-208-AWMA. Proceedings Annual Meeting, Air and Waste Management Association, Calgary. June 2010.

Kline, L.J., D.D. Davis. J.M. Skelly, and D.R. Decoteau 2009. Variation in ozone sensitivity within Indian Hemp and common milkweed selections from the Midwest. Northeastern Naturalist 16:307-313.

Papinchak, H.L., E.J. Holcomb, T.O. Best and D.R. Decoteau. 2009. Effectiveness of houseplants in reducing the indoor air pollutant ozone. HortTechnology 19:286-290.

Szantoi, Z., A.H. Chappelka, R.B. Muntifering, G.L. Somers. 2009. Cutleaf coneflower (Rudbeckia laciniata L.) response to ozone and ethylenediurea (EDU). Environ. Pollut. 157: 840-846.

Temple, P.J., Grantz, D.A. 2010. Air Pollution Stress. Physiology of Cotton. Editors: J. McD. Stewart, D. Oosterhuis, J.J. Heitholt, J. Mauney. Springer. Chapter 15

Tu, C., FL Booker, KO Burkey and S. Hu. 2009. Elevated atmospheric CO2 and O3 differentially alter nitrogen acquisition in peanut. Crop Science 49:1827-1836.