Brief Summary of Minutes of Annual Meeting

Annual Meeting Date and Location: November 12-13, 2004 South Carolina AES host, Hyatt Regency Hotel, Greenville, SC Attending Members: Dr. Karen Koch Florida AES Dr. Robert Aiken Kansas AES Dr. Robert Houtz Kentucky AES Dr. Steve Long Illinois AES Dr. Steve Rodermel Iowa AES Dr. Jack Preiss Michigan AES Dr. Robert Spreitzer Nebraska AES Dr. Jeff Harper Nevada AES Dr. Larry Daley Oregon AES Dr. Brandon Moore South Carolina AES Dr. Forrest Chumley Admin. Advisor, Kansas AES Attending Guests Interested in NC-1-142 Membership: Dr. Robert Aiken Kansas AES Dr. Michael Salvucci Arizona USDA-ARS Dr. Jyan-Chyun Jang Ohio AES Dr. Glenda Gillaspy Virginia AES This year's meeting was called to order by Dr. Brandon Moore, Chair. Scientific discussions began during the previous evening among members present and continued during breaks, lunch, and later at dinner. Participants each gave a 25 min overview of their previous years accomplishments, with questions and comments following their presentation. The talks included also a group discussion with our Administrative Advisor, Dr. Forrest Chumley. Dr. Chumley complemented the NC-1-142 members on the quality of their science, and the positive interaction between members. He indicated that NC-1-142 as a regional project is known for the high caliber of research by its members, and pointed out that NC-1-142 is essentially a regional project that extends across the entire country. Dr. Chumley noted, however, that the time-frame of the meeting left little time for interactions given the number of presentations and the limit to two days (Friday-Saturday). He suggested that the members consider extending the meeting time to three days (Friday-Sunday) so that more exchange and interaction between members could occur outside the framework of formal research presentations. He noted there is also an upcoming mid-project review to be considered in the near future. There was general agreement from the members that a more work-shop style approach should be considered for future meetings. Dr. Chumley also pointed out that it might be advantageous to consider a more geographically central location for the meetings, since this would help with travel time and there are members on both coasts. Dr. Chumley also addressed a previous suggestion at the last meeting that membership and attendance should be linked. Dr. Chumley pointed out that while many members receive financial support from their home institutions experiment station, this is not true for all members. Thus, travel costs for some are the individuals responsibility. The members were appreciative of Dr. Chumleys comments and presence at the meeting. Most discussion addressed Objectives 2, 3, and 4, which respectively are Photosynthetic Capture and Photorespiratory Release of CO2,Mechanisms Regulating Photosynthate Partitioning and Developmental and Environmental Limitations to Photosynthesis. Each of these objectives was well represented by the attendees at this year's meeting, but all members were involved in general discussion of the presentations. The business meeting was called to order after the talks. The meeting was attended by all members listed and lasted about 30 min. The primary issues were to 1) discuss candidates for incoming membership, 2) elect new officers, and 3) further consider our discussion with Dr. Chumley. The Chair noted that several previous members had resigned this year from the group and most members agreed that recruitment of strong new members is important to the vitality of the project. Four guests were considered for membership in NC-1-142, Robert Aiken, Michael Salvucci, Jyan-Chyun Jang, and Glenda Gillaspy. In addition to their oral presentations, all had provided the group with a c.v. and a statement of research interests as related to the NC-1-142 project. After some discussion including submitted comments from members not in attendance, a motion was made and seconded to extend membership to all four candidates. The motion was unanimously approved. New officers were selected with Dr. Karen Koch as Chair and host for the 2005 meeting, Dr. Robert L. Houtz as vice-chair, and Dr. Robert Spreitzer as secretary. The group agreed to maintain the current relative meeting date, 2 weekends before Thanksgiving. At the discretion of the new chair, next years meeting format might be altered along lines suggested by Dr. Chumley. Key discussions at the meeting: These centered on the science outlined in reports from the attending AES/ARS representatives, the visiting candidates for new membership, and the comments by Dr. Chumley. The primary issues of the business meeting are enumerated above. Assigned responsibilities: As decided by NC-1-142 participants, the leadership succession will progress from Dr. Brandon Moore (South Carolina AES) departing Chair of the 2004 meeting, to Dr. Karen Koch (Florida AES) incoming Chair and host for the 2005 meeting. Dr. Robert Houtz will become vice-chair and will host the 2006 meeting. Dr. Robert Spreitzer will serve as secretary for the 2005 meeting. As noted above, the weekend selected for the 2005 meeting again will be ONE WEEK EARLIER than in past years, now two weekends prior to the Thanksgiving holiday. The date will be November 11-13, 2005.

Photosynthesis is the fundamental process that enables the synthesis of sugars and all other biomolecules required for plant growth. Photosynthesis uniquely integrates light photochemistry, biochemical fixation of atmospheric carbon dioxide, and the utilization of inorganic nutrients to sustain and direct crop production under a variety of environmental conditions. This regional research project is an integrated cooperative program in which the expertise of AES and ARS investigators is applied to four aspects of the photosynthetic process. Objective 1: Photochemistry and the biogenesis of the photosynthetic apparatus Cooperating AES and/or ARS-USDA: IA (Rodermel). The molecular and biochemical coordination between nuclear and plastid gene expression continues as a major research focus by IA-AES (Rodermel), in which they are exploiting a number of available variegated plant mutants with green- and white-sectored leaves. These are proving useful to understanding chloroplast biogenesis, which is deficient in white sectored-leaves. For example, the GHOST variegation mutant of tomato was found to encode a chloroplast quinol oxidase that affects the biogenesis of chloroplasts and chromoplasts, as well as pericarp tissue morphogenesis, during the ripening process. Mutations in certain Arabidopsis chloroplast metalloproteases, FtsH5 (var1) and FtsH2 (var2), give rise to white sectors likely through a compromised post-transcriptional regulatory process. To gain further insight to the mechanism, a suppressor screen of var2 yielded several normal appearing lines. Map-based cloning revealed that suppression of variegation in one line is due to a splice site mutation in ClpC2, a chloroplast Hsp 100 chaperone. Further insight into the function of var2 in photosynthesis, plant development and plant stress responses might lead to the design of strategies to manipulate the photosynthetic capacity and quality of crop plants. Objective 2: Photosynthetic capture and photorespiratory release of CO2 Cooperating AES and/or ARS-USDA: IL (Portis), KY (Houtz), MO (Randall), NE (Chollet, Spreitzer), NV (Cushman),WA (Edwards, Okita). IL-ARS (Portis) has done experiments to better define the interactions between Rubisco and activase, their role in the inhibition of photosynthesis by high temperatures, and to develop alternative strategies for genetic engineering of Rubisco. A long-term collaboration with NE-AES (R. Spreitzer) uses chimeric spinach-tobacco activase proteins and site-directed mutagenesis to determine the specificity of activase for Solanaceae versus non-Solanaceae Rubiscos. In the current period, they showed that amino acids in the 267-339 region are responsible for Rubisco discrimination by tobacco activase, with residue 311 playing a major role. The Portis lab also showed that as temperature increases, Rubisco activity declines rapidly and certain inhibitors from the oxygenase reaction are formed more rapidly, but bind less tightly. Understanding activase function at high temperature will be important for crop improvement. Lastly, in collaboration with Dr. Henry Daniells laboratory (Univ. of Central Florida), in an exciting result, transgenic tobacco plants were made by chloroplast transformation of nuclear rbcS into an rbcS anti-sense plant. Progeny expressed mRNA from the introduced gene at high levels in the chloroplast, expressed Rubisco at almost the original wildtype level, and exhibited nearly restored photosynthesis rates. These results have opened an avenue for using chloroplast engineering for the evaluation of foreign Rubisco genes in planta. Studies at NE-AES (Spreitzer) continue to exploit Chlamydomonas rheinhardtii as a model system to use classical and molecular genetic strategies to define structure-function relations of Rubisco, with an overall goal to create a catalytically improved protein. Contemporary research has shown that this goal does require an in-depth understanding of these relationships. In an ongoing collaboration with Dr. Inger Andersson (Swedish Agricultural University, Uppsala), X-ray crystal structures have now been solved for more than 12 mutant Rubisco enzymes. Analysis of these has revealed novel insights and in some cases have shown that specific amino acids can have multiple functions. For example, the active-site Lys-175 was found to have an essential role in both catalysis and in vivo holoenzyme stability. Another finding was that the L290F mutant enzyme has a low CO2/O2 specificity and does not form a key H-bond with a substrate analog. Interestingly, this H-bond is restored in a L290F/A222T suppressor mutant. Since more than 2000 rbcL sequences are now available, bioinformatics approaches also are being used to define the structural basis for catalytic diversity among organisms. Making phylogenetic directed mutations has revealed novel properties at the small and large subunit interface that modulate catalytic properties and warrant further examination as possible targets for genetic engineering. Ongoing research at NE-AES (Chollet) has focused on the characterization and regulation of phosphoenolpyruvate (PEP) carboxylase isoforms in green algae, leaves and root nodules. In the current period, they have identified two novel PEPC genes in C. reinhardtii. These both lack an N-terminal phosphorylation domain present in green plant PEPC, yet characteristically are absent in prokaryotic PEPCs. Both genes were cloned and expressed in E. coli, and transcript abundance and biochemical activity were characterized. Work also was reported on the role of the C-terminal tetrapeptide in catalysis by sorghum PEPC. Site-directed mutagenesis on G961 showed this residue is critical for catalytic efficiency, but a G961A substitution was able to complement a related E. coli mutant. The Houtz laboratory (KY-AES) is characterizing chloroplast-localized post- and co-translational protein processing enzymes with emphasis on the large (LS) and small (SS) subunits of Rubisco. In Rubisco, a conserved amino acid sequence in the N-terminal region of the LS from Ala-9 to Lys-14 establishes the specificity for methylation at Lys-14. This region fits into a narrow cleft in Rubisco LS eN-methyltransferase (LSMT). Proof of principle was shown also by making an alternate substrate for Rubisco LSMT after fusing the N-terminal sequence from the LS to the N-termini of human carbonic anhydrase. In a separate project, possible cotranslational chloroplast protein processing by deformylases has been further examined. Deformylases DEF1 and DEF2 were shown to be targets of the inhibitor actinonin. This peptidomimetic antibiotic is known to inhibit chloroplast D1 polypeptide levels, leading to a disassembly of the photosystem II complex. The results confirm that N-terminal deformylation is an essential step in the accumulation and assembly of PSII subunit polypeptides in the chloroplasts of vascular plants. The mitochondrial pyruvate dehydrogenase complex (mtPDC) links glycolysis to the Krebs cycle by catalyzing the oxidative decarboxylation of pyruvate to acetyl-CoA. MO-AES (Randall) continues to examine the reversible phosphorylation of mtPDC, which provides the cardinal regulatory mechanism for the interaction of respiratory and photorespiratory metabolism. The protein kinase that phosphorylates mtPDC is a novel Ser-protein kinase, which they have now shown is a unique type of protein kinase that has a His-kinase like sequence, but Ser-kinase activity. Details of the phosphorylation mechanism have been better defined by kinase site directed mutagenesis, demonstrating residues critical for formation of the substrate ternary complex. A recombinant in vitro system also was developed to demonstrate that plant mtPDC has only one regulatory phosphorylation site on E1, unlike mammalian PDC. A reverse genetics approach has begun to establish permissible knockout lines of Arabidopsis PDK and PDC subunits. PDK and E1 knockout lines are being selected for homozygosity. NV-AES (Cushman) has continued study of the common ice plant, Mesembryanthemum crystallinum as a model for understanding Crassulacean acid metabolism (CAM). Using a molecular genetic approach, they characterized 9,733 expressed sequence tags (ESTs) from cDNAs derived from leaf tissues of well-watered and salinity-stressed plants. From these, 3,676 tentative unique gene sequences allowed rapid discovery of both known and unknown genes related to CAM induction and salinity tolerance in M. crystallinum. For example, the frequency of ESTs encoding light-harvesting and photosystem complexes and C3 photosynthetic enzymes decreased four- to seven-fold following salinity stress, while transcripts encoding CAM-related enzymes, pathogenesis-related, senescence-associated, cell death-related, and stress-related proteins such as heat shock proteins, chaperones, early light-inducible proteins, ion homeostasis, antioxidative stress, detoxification, and biosynthetic enzymes for osmoprotectants increased 2-12-fold in salt stressed plants. In a separate project, the Cushman lab has examined many of the molecular components and their transcript expression in relation to circadian clock function in M. crystallinum. While some differences were noted in the diurnal expression profiles of particular components when compared to Arabidopsis, the clock in M. crystallinum likely functions in a very similar fashion to the clock in Arabidopsis. This indicates that such a clock could control CAM without requiring additional components of the central oscillator. Objective 3: Mechanisms regulating photosynthate partitioning Cooperating AES and/or ARS-USDA: FL (Koch), IA (Knapp, Rodermel), IL (Bohnert, Huber), MI (Preiss, Loescher),NE (Chollet), NV (Harper), PA (Guiltinan), SC (Moore), WA (Edwards, Okita),WI (Duke). FL-AES (Koch) has examined the response of an Arabidopsis vacuolar invertase gene family to sugar and ABA. Since sucrose is the major carbon and energy source for most importing plant cells, effects of plant invertases on hexose/sucrose balance can markedly alter expression of diverse, sugar-regulated genes. In Arabidopsis, there are two vacuolar invertase genes, plus six cell wall invertase genes. In the current period, methods were developed for quantitative Q-RT-PCR and gene-specific amplicons for each family member. Tissue specific gene expression and gene-specific regulatory effects of sugar and ABA were demonstrated. These contrasting responses provide a mechanism for maximizing flexibility and balance at a key point in both sucrose metabolism and sugar/ABA signaling. Effects of knock-outs in these invertases is also being investigated in conjunction with IA-AES (S. Rodermel). The Koch lab also has been involved in identifying and characterizing maize knockout mutants. For example, vegetative phenotypes were examined for maize mutants lacking one or two sucrose synthases. Differential and additive affects were noted, especially with regard to seed weight and germination. To identify further maize knockout mutants, reverse-genetics grids are being constructed using a phenotypically uniform inbred population (UniformMu). Initial screening is in progress for knockouts in key genes for cell wall biosynthesis. IL-ARS (Huber) has done novel experiments probing the control of plant metabolism by protein phosphorylation. In one approach, far-Western overlays of extracts prepared from light- or dark-treated spinach (Spinacia oleracea) leaves were used to identify factors that affected binding of cellular proteins to labeled 14-3-3 protein. Binding of phosphorylated proteins was shown to be stimulated by polycations, but specificities were not notably affected. Using recently obtained modification-specific antibodies, IL-ARS has begun to monitor the phosphorylation status of nitrate reductase (NR) in Arabidopsis leaves by immunoblotting. In collaboration with Dr. Mike Sussman (UW-Madison), they are screening CDPK-knockout lines to identify the requisite NR-kinase(s). Also important will be identifying the factors that 'trigger' the release of bound 14-3-3 protein and thereby the activation and dephosphorylation of NR. In conjunction with FL-AES (K. Koch), IL-ARS also is characterizing stable maize transgenics that express wild type and phosphorylation site mutants of the SUS1 (sucrose synthase) protein. To aid these studies, they have obtained antibodies that specifically detect the SUS3 protein, which has not been previously characterized at the protein level. In collaboration with SC-AES (B. Moore), IL-ARS has initiated a new project to determine whether HXK-mediated glucose signaling is affected by nitric oxide (NO). It is known that glucose inhibition of Arabidopsis seedling growth and greening can be antagonized by nitrate. This effect may be mediated by NO (known to be generated by reduction of nitrite by NR), because HXK1 contains 3 cysteine residues in a consensus motif predicted to be susceptible to S-nitrosylation. MI-AES (Preiss) have done studies to determine structure-function aspects of both the ADPglucose pyrophosphorylase (AGP) and glycoge synthase. AGP catalyzes the first committed and rate-limiting step in starch biosynthesis in plants and glycogen synthase similarly for glycogen biosynthesis in bacteria. They are the master regulators of storage polysaccharide accumulation in plants and bacteria, being allosterically activated or repressed by metabolites of energy flux. In a significant development, they report the first atomic resolution structure of an AGP crystallized from potato tuber. Communication between the regulator binding sites and the active site of AGP are both subtle and complex, and involve several distinct regions of the enzyme including the N-terminus, the glucose-1-phosphate binding site and the ATP binding site. These structures provide insights into the mechanism for catalysis and allosteric regulation of AGP. MI-AES also modeled the E. coli glycogen synthase catalytic site based on three glycosyltransferases with a GT-B fold. The model was validated after analysis of targeted site-directed mutants which had predictable effects on reaction kinetic constants. The Harper lab (NV-AES) has just recently joined in the project objective. They have a long-standing research interest in understanding and modifying the transport machinery that moves sucrose and other nutrients from source to sink. The driving force for solute transport across plant cell plasma membranes is a proton gradient produced by a P-type H+-ATPase. Proton pumps are responsible for the driving force underlying sucrose loading and its long distance transport through phloem. Harper was the first to clone a plant proton pump in 1989, and has pursued an active research program to understand the structure and biological functions of these pumps. In the past year, SC-AES (Moore) has examined the cellular process by which Arabidopsis hexokinase1 (HXK1) transduces a sugar signal. HXK-dependent glucose signaling affects plant growth throughout the plant life cycle and modulates a number of genes related to carbohydrate synthesis and utilization. Using leaf fractionation and bioimaging of 35S::HXK1-GFP plants, HXK1 was shown to occur exclusively at the mitochondria under varying treatments and in all tissues. Thus, glucose signal transduction might involve mitochondria translocation to or from the nucleus, or might involve a mobile HXK1 interacting protein. A previous proteomics experiment identified vegetative actin associated with leaf HXK1. In collaboration with Dr. Richard Meagher (University of Georgia), these scientists used actin mutants and pharmacological treatments of wildtype plants to demonstrate a key finding that the actin cytoskeleton has a direct role in HXK1-dependent glucose signaling. In collaboration with IL-ARS (S. Huber), they have initiated experiments to determine if and to what significance that AtHXK1 might be nitrosylated. Nitric oxide is a recognized intracellular signaling molecule that mediates plant responses to hormones and pathogens. Initial experiments using an HXK1-null mutant and wildtype plants, indicates that HXK1 is required for certain standard seedling responses to NO. Furthermore, HXK1 activity was partly repressed by incubation with an NO donor molecule, thus supporting the possibility that HXK1 is directly modified by NO. Research on starch biosynthesis by WA-AES (Edwards and Okita) is directed at understanding its significance in source (leaves) and sink (seed) tissues to photosynthesis, plant growth and crop production. Current work with Arabidopsis has been directed at complementing the TL46, starch-deficient mutant with altered forms of the large subunit of ADP-glucose pyrophos-phorylase (AGP). These forms are less sensitive to inhibition by inorganic phosphate. Transformants have higher AGPase activity and higher starch synthesis than wild type plants. Backcrosses of these up-regulated transformants are being made to select for stable lines with high AGPase activity and starch synthesis. Current work with rice has resulted in the exciting finding that several lines which express potato AGPase exhibit elevated leaf starch levels, higher productivity, and increased yields in field plots at the RDA, Suweon, Korea (in collaboration with Dr. Jong-Sug Park). These results support the view that increases in leaf starch provide, at the minimum, additional carbon and energy reserves during the night, which increase plant growth and development. Objective 4: Developmental and environmental limitations to photosynthesis Cooperating AES and/or ARS-USDA: Guam (Marler), IL (Below, Bohnert, Long), KS (Knapp), MI (Loescher), MN (Jones),NE (Markwell), NV (Cushman, Harper, Mittler), OR (Daley), WA (Edwards, Okita). IL-AES (Below) has examined an important question of how N assimilates move through the young maize earshoot during seed development. To do this, they assessed amino acid metabolism in a 2 year field study, examining cob and spikelet tissues during the critical two weeks following silking. Amino acid profiles and enzyme activities were determined in specific reproductive organs in two maize hybrids grown at two levels of supplemental N fertilizer. Most amino acids except Gln accumulated over time in unpollinated spikelets and cobs, and nitrogen supply had a variable effect on individual amino acid levels in young cobs and spikelets. Interestingly, IL-AES showed that the cob performs significant enzymatic interconversions among Gln, Ala, Asp and Asn during early reproductive development, which may precondition the N assimilate supply for sustained kernel growth. Furthermore, the measured amino acid profiles and enzymatic activities suggest that the Asn/Gln ratio in cobs may be part of a signal transduction pathway that indicates plant N status for kernel development. IL-AES (Long) has continued their long-term interests in the acclimation of photosynthesis to global change, primarily the direct effects of rising tropospheric concentrations of CO2 and ozone. A major focus over the past two years has been examining the interactive effects of predicted 2050 levels of ozone and CO2 on soybean (also Arabidopsis) under fully-open air field conditions using the SoyFACE facility. In agreement with the 2003 experiment, their larger scale 2004 FACE experiment has revealed that while rising CO2 increases photosynthesis and yield, the increases even in a nitrogen fixing crop are smaller than anticipated, while the simultaneous increase in ozone results in a substantial yield loss. In the 2004 experiment, a 50% increase in CO2 increased yield by almost 20%, but increased ozone decreased yield by 20%. Ozone decreased photosynthesis, accelerated senescence and crop maturation, and resulted in fewer pods, fewer seeds per pod and smaller seeds. Although not fully analyzed, it is clear that elevation of CO2 partially offsets the damage caused by elevated ozone. However, these results do suggest that the prediction that the detrimental effects of global change in the corn belt may be offset by increases due to rising CO2 may be over-optimistic. IL-AES also has developed improved modeling temperature functions for predicting photosynthesis. These will provide a basis for the development of better mechanistic models of crop responses to global climate change KS-AES (Knapp and associates) have pursued studies of plant and ecosystem responses to altered precipitation and temperature. The sensitivity of native grasslands to predicted changes in temperature and rainfall variability poses important questions for the future productivity and sustainability of these ecologically and agriculturally important ecosystems. Results from an ongoing field experiment in which precipitation patterns are being manipulated, indicate that predicted changes in rainfall timing can alter plant processes, from leaf-level photosynthesis to whole plant productivity. In particular, prolonged inter-rainfall droughts during the growing season reduce the potential for recovery of leaf-level photosynthesis once a rewetting rainfall event occurs. KS-AES also showed that plant responses to a more variable rainfall regime could have significant effects on soil processes and ecosystem-level C cycling and storage. Increased temperatures, another prediction of climate change models, may increase plant water stress even further and amplify the effects of increased soil moisture variability. In another project, KS-AES is evaluating photosynthetic performance and thermotolerance in selected recombinant inbred lines of sorghum. The goal is to identify trait differentiation between tolerant and heat-susceptible lines in order to map the relative positions of thermotolerance traits in the sorghum genome. MI-AES (Loescher) has collaborated with WA-AES (Edwards, Okita) in studies on salt tolerance using Arabidopsis as a model plant. Arabidopsis transformed with a celery gene for synthesis of mannitol (mannitol 6-P reductase), produces mannitol and has higher growth tolerance to salinity than wildtype Arabidopsis, which does not produce mannitol. In current work, these scientists have shown that transformants were more tolerant to salinity as shown through analysis of photosynthesis and a number of growth parameters. Publication of a joint paper on this work will be forthcoming. NE-AES (Markwell) has further examined the kinetics and substrate specificity of Arabidopsis leaf formate dehydrogenase. This enzyme catalyzes the oxidation of formate to CO2 in plant mitochondria. Experiments indicated that anomalous reaction kinetics might be due to a latent capacity for alternate substrate utilization which is realized upon heating. One possibility is that heating generates a molten globule form of the enzyme which upon cooling is able to refold in an alternate conformation that reveals a new substrate binding ability. In the past year, these scientists also have initiated the directed evolution of FDH to obtain variants with increased substrate affinities. A PCR based method of random mutagenesis has been developed and library screening will commence in the following year. NE-AES also has further examined the reversible denaturation of plant proteins, using as a model system the denaturation and renaturation of soybean Kunitz trypsin inhibitor (SKTI). This 21.5-kDa protein is a food allergen, and is unusually resistant to thermal and chemical denaturation. Spectroscopic and biochemical analyses indicated that during denaturation, SKTI shows major structural changes, possibly having formed a molten globule structure as a partially unfolded intermediate. NV-AES (Cushman) has further examined the molecular genetic basis for salinity and drought stress limitations on plant photosynthesis. In the past year, they have investigated a salinity- and dehydration-stress responsive calcium-dependent protein kinase (CDPK) isolated from Mesembryanthemum crystallinum (McCPK1). CDPKs are sensor-transducer proteins capable of decoding calcium signals in diverse phosphorylation-dependent calcium signaling networks, including signals generated by environmental stresses. Recombinant McCPK1 was shown to catalyze Ca-dependent substrate phosphorylation, undergo autophosphorylation, and in response to decreased humidity undergo altered sub-cellular localization. In a related project, these scientists have used a series of yeast two-hybrid screens to identify potential substrates of a water deficit- and salt stress-inducible CDPK from Arabidopsis (AtCPK11). Several interacting proteins were identified including likely transcriptional regulators, some of which co-localized in the nucleus with AtCPK11 and showed coordinated up-regulation induced in seedlings by water-deficit stress. NV-AES (Harper) also has an important practical goal to identify potential mechanisms to improve crop yield. This includes identifying genes that affect a plants response to biotic and abiotic stresses (including light stress), as well as developmental timing and plant structure. For example, a current project funded in collaboration with J. Cushman and R. Mittler (NV-AES) is to screen Arabidopsis mutants to identify genes that affect a plants ability to grow under extreme environments. These genes will provide important leads in the potential engineering of higher yield crops. In a more global context, it is clear that phospho-signaling provides regulatory switches that affect all aspects of plant growth and response to the environment. The Harper lab has been working on the structure and biological functions of CDPKs since. NV-AES (Mittler) is interested in understanding the regulation and function of the plant reactive oxygen gene network. In Arabidopsis, at least 152 genes are involved in managing the level of reactive oxygen species (ROS) in cells and many questions remain unanswered in regard to their regulation and protective roles. In leaf cells, the chloroplast is considered to be a focal point of ROS metabolism. It is a major producer of O2- and H2O2 during photosynthesis and it contains a large array of ROS-scavenging mechanisms that have been extensively studied. In contrast, the function of the cytosolic ROS-scavenging mechanisms of leaf cells is largely unknown. In the past year, NV-AES has used targeted knockout plants to demonstrate that the cytosol has a key role in protecting the chloroplast during light stress. Further work has characterized the signaling events that ensue in response to a moderate level of light stress in plants deficient in this major cytosolic ROS-scavenging mechanism. OR-AES (Daley) has pursued examination of the polyphyletic evolution of CAM by considering operational definitions of evolution, development, and manifestation of CAM. Project Participant Plans for 2004-2005 FL-AES will extend analysis of invertase gene responses and roles in sugar/ABA signaling in knockout mutants of both maize and Arabidopsis (in collaboration with IA-ARS). They also will continue to screen the UniformMu, reverse genetic grids for knockouts in genes of polysaccharide biosynthesis (with preference to cell wall and kernel targets). IA-AES will continue studies to examine the role of VAR2 in photosynthesis by focusing on second-site suppressor studies of var2. IL-ARS will collaborate with NV-AES to identify synthetic peptide inhibitors for CDPKs based on both the classical and recently identified non-classical phosphorylation motifs. IL-ARS also will characterize transgenic Arabidopsis plants over-expressing 14-3-3s, wild type and cation-activation mutants, to test several postulates concerning the role of 14-3-3s in regulation of nitrate reductase activity and steady-state protein level. They will continue collaboration with FL-AES to characterize transgenic corn plants expressing phosphorylation site mutants of sucrose synthase. IL-AES will integrate their metabolite profiling findings with mRNA expression profiling and genetic mapping in maize reproductive tissues to discover the metabolic pathways and genes that are associated with N use efficiency. Also, IL-ARS will continue (in collaboration with NE-AES) studies of the interaction of activase with Rubisco by analysis of Chlamydomonas site-directed large subunit mutants and site-directed mutants of activase proteins. And, IL-ARS will continue collaboration with AZ-ARS to examine the effects of high temperature on inhibitor formation during Rubisco catalysis and the activation of Rubisco by activase. KS-AES will collect new genomic and plant physiological data to determine how gene function and physiological response is altered by our rainfall and temperature manipulations. Ongoing studies by KY-AES are aimed at identifying other LSMT substrates and the general mechanism for recognition and methylation of target protein lysyl residues by other protein N-methyltransferases. MI-AES is making a major effort to obtain crystals and three-dimensional structures of the E. coli and potato tuber ADPGlc PPases and of the E. coli glycogen synthase. MO-AES will characterize PDK and E1alpha knockouts under various physiological conditions and stresses, and determine if the different mitochondrial E2 genes code proteins with specific functions for PDC. NE-AES will further characterize PEPC from C. reinhardtii using an RNAi approach, and will examine the role of N959 in the inhibitor binding site. NV-AES will determine if a plasma membrane calcium pump isoform, ACA12, can be used to increase pathogen resistance and will screen mutant plants for stress sensitive phenotypes resulting from gene knockouts in stress-response genes. NV-AES also will continue their functional analysis of CDPKs and closely related protein kinases from Arabidopsis thaliana. Protein-protein interaction studies will be expanded to include both targeted and random yeast two-hybrid screens of other unknown gene products regulated by abiotic stress. SC-AES will examine the link between HXK signaling, the actin cytoskeleton, and mitochondrial function. They also will further examine (in collaboration with IL-ARS) interactions between NO and HXK-dependent signaling processes. WA-AES will repeat field trials this year with transgenic rice, using a much larger scale planting. Plants will be assessed for physiological growth characteristics and leaf starch metabolism. They also will evaluate the potential role of leaf starch serving as a transient sink to alleviate photosynthesis feedback.

Akhani, H, J Barroca, N Koteyeva, E Voznesenskaya, V Franceschi, G Edwards, SM Ghaffari, W Stichler, H Ziegler 200X) Bienertia sinuspersici (Chenopodiaceae): a new species from SW Asia and discovery of a third terrestrial C4 plant without Kranz anatomy. Systematic Botany, submitted. Aluru, M., S.R. Rodermel (2003) Identification of IMMUTANS as a plastid terminal oxidase: its role in differentiation, carotenoid biosynthesis and chlororespiration. Recent Res. Devel. Plant Mol. Biol. 1: 39-55. Aluru, M.R, S.R. Rodermel (2004) Control of chloroplast redox by the IMMUTANS terminal oxidase. (Refereed review). Physiologia Plantarum 120: 4-11 (Cover Article). AINSWORTH EA, DAVEY PA, HYMUS GJ, OSBORNE CE, ROGERS A, BLUM H, NOSBERGER J, LONG SP (2003) Is stimulation of leaf photosynthesis by elevated carbon dioxide concentration maintained in the long term? A test with Lolium perenne grown for 10 years at two nitrogen fertilization levels under Free Air CO2 Enrichment (FACE). Plant, Cell Environment 26: 705-714. AINSWORTH E.A., ROGERS A., BLUM H., NOSBERGER J., LONG S.P. (2003) Variation in acclimation of photosynthesis in Trifolium repens after eight years of exposure to Free Air CO2 Enrichment (FACE). J Exp Botany 54: 2769-2774. AINSWORTH E.A., ROGERS A., NELSON R. & LONG S.P. (2004) Testing the "source-sink" hypothesis of down-regulation of photosynthesis in elevated CO2 in the field with single gene substitutions in Glycine max. Agricultural and Forest Meteorology 122: 85-94. AINSWORTH EA, TRANEL PJ, DRAKE BG, LONG SP (2003) The clonal structure of Quercus geminata revealed by conserved microsatellite loci. Molecular Ecology 12: 527-532. Ballicora, M.A., A.A. Iglesias, J. Preiss (2004) ADP-glucose Pyrophosphorylase; a Regulatory enzyme for Plant Starch Synthesis. Photosynthesis Research, 79: 1-24. Barr, J., W.S. White, L. Chen, H. Bae, S. Rodermel (2004) The GHOST terminal oxidase regulates developmental programming in tomato fruit. Plant, Cell Environment 27: 840-852. (Cover article) Barroca, J, LR Murphy, VR Franceschi, R Lee, E Roalson, GE Edwards, MS Ku. (200X) Diversification and plasticity of C4 photosynthetic pathway in Eleocharis (Cyperaceae). In: Proceedings of the 13th International Photosynthesis Congress. Springer. In press. Baxter I, et al., J. Young, J.F. Harper (2004) Evidence for an endomembrane function of a plasma membrane proton pump. PNAS (in revision). Bejar, C.M., M.A. Ballicora, D.F. Gomez-Casati, A.A. Iglesias, J. Preiss (2004) The ADP-glucose pyrophosphorylase from Escherichia coli comprises two tightly bound distinct domains. FEBS Letters, 573: 99-104. Below FE, JR Seebauer, M Uribelarrea, MC Schneerman, SP Moose (2004) Physiological changes accompanying long term selection for grain protein in maize. Plant Breed. Rev. 24(1):133-151. Bergerou JA, LE Gentry, MB David, FE Below (2004) Role of N2 fixation in the soybean N credit in maize production. Plant and Soil 262:383-394. BERNACCHI C.J., CALFAPIETRA C., DAVEY P.A., WITTIG V.E., SCARASCIA-MUGNOZZA G.E., RAINES C.A., LONG S.P. (2003) Photosynthesis and stomatal conductance responses of poplars to free-air CO2 enrichment (PopFACE) during the first growth cycle and immediately following coppice. New Phytologist 159: 609-621. BERNACCHI C.J., MORGAN P.B., ORT D.R., LONG S.P. (2004) The growth of soybean under free air [CO2] enrichment (FACE) stimulates photosynthesis while decreasing in vivo Rubisco capacity. Planta 219 (in press). BERNACCHI CJ, PIMENTEL C, LONG SP (2003) In vivo temperature response functions of parameters required to model RuBP-limited photosynthesis. Plant, Cell Environment 26: 1419-1430. Boxall SF, Foster JM, Bohnert HJ, Cushman JC, Nimmo HG, Hartwell J. (2005) Conservation and divergence of the central circadian clock in the stress-inducible CAM plant Mesembryanthemum crystallinum: clock operation in a CAM halophyte reveals clock compensation against abiotic stress. Plant Physiol. In press. Chehab EW, Patharkar OR, Hegeman AD, Taybi T, Cushman JC. (2004) Autophosphorylation and subcellular dynamics of a salt- and water deficit stress-induced calcium-dependent protein kinase from Mesembryanthemum crystallinum. Plant Physiol. 135: 1430-1446. Christodoulou, J., A. Malmendal, J.F. Harper, W.J. Chazin (2004) The N-terminal lobe of the CaM-like domain functions as the Ca2+ Sensor for the activation of a calcium-dependent protein kinase (CDPK) from Arabidopsis J. Biol Chem 279:29092-100. Cross, J.M., M. Clancy, J.R. Shaw, S. Boehlein, T.W. Greene, R.R. Schmidt, T.W. Okita and L.C. Hannah (2004) A polymorphic motif in the small subunit of ADP-glucose pyrophosphorylase modulates interactions between the small and large subunits. Plant J. In Press. Cross, JM, M. Clancy, J.R. Shaw, T.W. Greene, R.R. Schmidt, T.W. Okita, L.C. Hannah (2004) Both subunits of ADP-glucose pyrophosphorylase are regulatory. Plant Physiol. 135:137-144. Cruz JA, TJ Avenson, A Kanazawa, K Takizawa, GE Edwards, DM Kramer (200X) Plasticity in light reactions of photosynthesis from energy production and photoprotection. J Exp Bot., In Press. DAVEY P, HUNT S, HYMUS G, DRAKE B, DELUCIA E, KARNOSKY, D, LONG, SP (2004) Respiratory oxygen uptake is not decreased by an instantaneous elevation of [CO2], but is increased by long-term growth in the field at elevated [CO2]. Plant Physiology 134: 520-527. Davletova, S., L. Rizhsky, H. Liang, Z. Shengqiang, D.J. Oliver, J. Coutu, V. Shulaev, K. Schlauch, R. Mittler (2004) Cytosolic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of Arabidopsis. Plant Cell, in press. Dhingra, A., Portis A.R. Jr, Daniell H. (2004) Enhanced translation of a chloroplast-expressed RbcS gene restores small subunit levels and photosynthesis in nuclear RbcS antisense plants. Proc. Natl. Acad. Sci. USA 101:6315-6320. Dinkins, R.D., H.M. Conn, L.M.A. Dirk, M.A. Williams, R.L. Houtz (2003) The Arabidopsis thaliana peptide deformylase 1 protein is localized to both mitochondria and chloroplasts. Plant Sci 165: 751-758. Du, Y.C., Peddi S.R., Spreitzer R.J. (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/ oxygenase. J. Biol. Chem. 278: 49401-49405. Edwards GE, VR Franceschi, EV Voznesenskaya (2004) Single-cell C4 photosynthesis versus the dual-cell (Kranz) paradigm. Annual Review of Plant Biology 55: 173-196. Fay, P.A., A.K. Knapp, J.M. Blair, J.D. Carlisle, J.K. McCarron, B.T. Danner (2003) Rainfall timing, soil moisture dynamics, and plant responses in a mesic tallgrass prairie ecosystem. Pages 147-163 In Changing Precipitation Regimes and Terrestrial Ecosystems. A North American Perspective. (J.F. Weltzin and G.R. McPherson, eds.) University of Arizona Press. Hardin, S.C., S.C. Huber (2004) Proteasome activity and the post-translational control of sucrose synthase stability in maize leaves. Plant Physiol. Biochem. 42: 197-208. Hardin, S.C., H. Winter, S.C. Huber (2004) Phosphorylation of the amino-terminus of maize sucrose synthase in relation to membrane association and enzyme activity. Plant Physiol. 134: 1427-1438. Harper, C.W., J.M. Blair, P.A. Fay, A.K. Knapp, J.D. Carlisle (200X) Increased rainfall variability and reduced rainfall amount decreases soil CO2 flux in a grassland ecosystem. Global Change Biology, In press. Harper J, G Breton, A Harmon (2004) Decoding CA signals through plant protein kinases Annual Reviews of Plant Biology. 55: 263-88. Hegeman A.D., A.C. Harms, M.R. Sussman, A.E. Bunner, J.F.Harper (2004) An isotope labeling strategy for quantifying the degree of phosphorylation at multiple sites in proteins. J Am Soc Mass Spectrom 15: 647-53. Hirani, T.A., Tovar-Mendez A., Miernyk J.A., Randall D.D. (2004). Asp-295 is important for phosphorylation of the pyruvate dehydrogenase E1-alpha active-site loop by pyruvate dehydrogenase kinase. Submitted-in revision. Houtz, R.L., A.R. Portis Jr. (2003) The life of ribulose-1,5-bisphosphate carboxylase/oxygenase  post-tranlational facts and mysteries. Minireview. Arch Biochem Biophys 414: 150-158 (special issue on C-fixing enzymes). Huber, S.C. S.C. Hardin (2004) Numerous post-translational modifications provide opportunities for the intricate regulation of metabolic enzymes at multiple levels. Current Opinion Plant Biology 7: 318-322. Hussain,D., M.J. Haydon, Y. Wang, E. Wong, S.M. Sherson, J. Young, J. Camakaris, J.F. Harper, C.S. Cobbett (2004) P-type ATPase heavy metal transporters with roles in essential zinc metabolism in Arabidopsis. Plant Cell 16: 1327-1339. Hwang, S.-K., P.R. Salamone, H.Kavakli, C.J. Slattery, T.W. Okita (2004) Rapid purification of the potato ADP-glucose pyrophosphorylase by poly-histidine mediated chromatography. Protein Purif. Express. 38: 99-107. Ivanov B, K Asada, D Kramer, GE Edwards (200X) The properties of photosynthetic electron transport in bundle sheath cells of maize. Ascorbate can effectively charge cyclic electron flow. Planta, In Press. Karkehabadi, S., Taylor, T.C., Spreitzer, R.J., Andersson, I. (2004) Altered intersubunit interactions in crystal structures of catalytically-compromised ribulosebisphosphate carboxylase/oxygenase. Biochemistry (in press). Kim, K., Portis, A.R. Jr. (2004) Oxygen-dependent H2O2 production by Rubisco. FEBS Letters 571: 124-128. Koch, K.E. (2004) Sucrose metabolism: Regulatory mechanisms and pivotal roles in sugar sensing and plant development. Current Opin Plant Biol 7: 235-246. Kore-eda S, Cushman MA, Akselrod I, Bufford D, Fredrickson M, Clark E, Cushman JC (2004) Transcript Profiling of Salinity Stress Responses by Large-Scale Expressed Sequence Tag Analysis in Mesembryanthemum crystallinum. Gene 341:83-92. Kore-eda S, Noake C, Ohishi M, Ohnishi J, Cushman JC (2005) Transcriptional regulation of organellar metabolite transporters during induction of crassulacean acid metabolism in Mesembryanthemum crystallinum. Functional Plant Biology. Submitted. Kramer DM, TJ Avenson, GE Edwards (2004) Dynamic flexibility in the light reactions of photosynthesis governed by both electron and proton transfer reactions. Trends Plant Science 9: 339-348. Kramer DM, TJ Avenson, GE Edwards (2004) Response to Johnson: Controversy remains: regulation of pH gradient across the thylakoid membrane. Trends Plant Science 9 571-572. Kramer DM, G Johnson, O Kiirats, GE Edwards (2004) New fluorescence parameters for the determination of QA redox status and excitation energy fluxes. Photosynthesis Res 79:209-218. Kramer DM, A Kanazawa, JA Cruz, B Ivanov, GE Edwards (200X) The relationship between photosynthetic electron transfer and its regulation. In: Govindjee, GC Papageorgiou, eds, Chlorophyll Fluorescence: The Signature of Green Plant Photosynthesis. The Netherlands: Kluwer Academic Publishers. In Press. LEAKEY A.D.B., BERNACCHI C.J., DOHLEMAN F.G., ORT D.R. & LONG S.P. (2004) Will photosynthesis of maize (Zea mays) in the US Corn Belt increase in future CO2 rich atmospheres? An analysis of diurnal courses of CO2 uptake under free-air concentration enrichment (FACE). Global Change Biology 10: 951-962. LONG SP, AINSWORTH EA, ROGERS A, ORT DR (2004) Rising Atmospheric Carbon Dioxide: Plants Face The Future. Annual Reviews of Plant Biology 55: 591-628. LONG S.P., BERNACCHI C.J. (2003) Gas exchange measurements, what can they tell us about the underlying limitations to photosynthesis? Procedures and sources of error. J Exp Botany 54: 2393-2401. Lonosky, P., X. Zhang, V. Honavar, D. Dobbs, A. Fu, S. Rodermel (2004) A proteomic analysis of maize chloroplast biogenesis. Plant Physiology 134: 560-574. Mamedov, T.G., Moellering E.R., Chollet R. (2004) Identification and expression analysis of two genes encoding novel and distinct molecular forms of eukaryotic PEP carboxylase in the green microalga Chlamydomonas reinhardtii. Plant J.. submitted. Mazarei, M., K.A. Lennon, D. P. Puthoff, S. R. Rodermel, T. J. Baum (2004) Homologous soybean and Arabidopsis genes share responsiveness to cyst nematode infection. Molecular Plant Pathology 5: 409-423. Mazarei, M., K.A. Lennon, D. P. Puthoff, S. R. Rodermel, T. J. Baum (2004) Expression of an Arabidopsis phosphoglycerate mutase homologue is localized to apical meristems, regulated by hormones, and induced by sedentary plant-parasitic nematodes. Plant Mol. Biol. 53: 513-530. Mittler, R., S. Vanderauwera, M. Gollery, F. Van Breusegem (2004) The reactive xxygen gene network of plants. Trends Plant Science 9: 490-498. MIYAZAKI S, FREDRICKSEN M, HOLLIS KC, POROYKO V, SHEPLEY D, GALBRAITH DW, LONG SP, BOHNERT HJ (2004) Transcript expression profiles of Arabidopsis thaliana grown under controlled conditions and open-air elevated concentrations of CO2 and of O3. Field Crops Research 90: 47-59. Moore B.d. (2004) Bifunctional and moonlighting enzymes: lighting the way to regulatory control. Trends Plant Science 9: 221-228. (cover article) MORGAN PB, AINSWORTH EA, LONG SP (2003) How does elevated ozone impact soybean? A meta-analysis of photosynthesis, growth and yield. Plant, Cell Environment 26: 1317-1328. MORGAN PB, BERNACCHI CJ, ORT DR, LONG SP (2004) An in vivo analysis of the effect of season-long open-air elevation of ozone to anticipated 2050 levels on photosynthesis in soybean. Plant Physiology 135, 2348-2357. MORGAN P.B., BOLLERO G., NELSON R.L., LONG S.P. (2005) Season-long elevation of ozone concentration by 20 % under fully open-air conditions decreases the growth and production of Midwest soybean crops by ca. 20 %. Environmental Pollution (in press). MORGAN P.B., BOLLERO G., NELSON R.L., DOHLEMAN F.G., LONG S.P. (2005) Smaller than predicted increase in above-ground net primary production and yield of field-grown soybean was found when [CO2] is elevated in fully open-air. Global Change Biology 11 (in press). NAIDU SL, LONG, SP (2004) Potential mechanisms of low-temperature tolerance of C4 photosynthesis in Miscanthus x giganteus: an in vivo analysis. Planta 219 (in press). NAIDU SL, MOOSE SP, AL-SHOAIBI AK, RAINES CA, LONG SP (2003) Cold tolerance of C4 photosynthesis in Miscanthus x giganteus: Adaptation in amounts and sequence of C4 photosynthetic enzymes. Plant Physiology 132: 1688-1697. Park, S., S. Rodermel (2004) Mutations in ClpC2/Hsp100 suppress the requirement for FtsH in thylakoid membrane biogenesis. Proc. Natl. Acad. Sci. USA 101: 12765-12770. Peddi, S.R., Karkehabadi, S., Anwaruzzaman, M., Andersson, I., Spreitzer, R.J. (2004) Evolutionary divergence in the structure of the small-subunit bA-bB loop of ribulose-1,5-bisphosphate carboxylase/oxygenase is not essential for assembly but influences large-subunit catalysis. In Photosynthesis: Fundamental Aspects to Global Perspectives, eds. A. van der Est and D. Bruce. Allen Press, Lawrence, Kansas (in press). Portis, A.R. Jr. (2004) Rubisco activase. Encylopedia of Plant and Crop Science, R.M. Goodman ed., Marcel Dekker, Inc, New York, NY, pp. 1117-1119. Preiss, J. (2004) The Biochemistry and Molecular Biology of Starch synthesis. IN Starch in Food. Woodhead Publishing Ltd, In Press. Puthoff, D.P., D. Nettleton, S.R. Rodermel, T.J. Baum (2003) Arabidopsis gene expression changes during cyst nematode parasitism revealed by statistical analyses of microarray expression profiles. Plant Journal 33: 1-11. Rodermel, S., S. Park. 2003. Pathways of intracellular communication: tetrapyrroles and plastid-to-nucleus signaling. BioEssays 25: 631-636. Rodriguez Milla MA, Uno Y, Townsend J, Maher E, Cushman JC (2005) Identification of potential substrates of AtCPK11, a calcium-dependent protein kinase induced by water-deficit and salt stress in Arabidopsis thaliana. Submitted. ROGERS A., ALLEN D.J., DAVEY P.A., MORGAN P.B., AINSWORTH E.A., BERNACCHI C.J., CORNIC G., DERMODY O., DOHLEMAN F.G., HEATON E.A., MAHONEY J., ZHU X.G., DELUCIA E.H., ORT D.R., LONG S.P. (2004) Leaf photosynthesis and carbohydrate dynamics of soybeans grown throughout their life-cycle under Free-Air Carbon dioxide Enrichment. Plant Cell Environment 27: 449-458. Roychaudhuri, R., Sarath G., Zeece M., Markwell, J. (2004) Stability of the allergenic soybean Kunitz trypsin inhibitor. Biochim. Biophys. Acta 1699: 207-212. Sakulsinghharoj, C., S.-B. Choi, S.-K. Hwang, G.E. Edwards, J. Bork, C.R. Meyer, J. Preiss, T.W. Okita (2004) Engineering starch biosynthesis for increasing rice seed weight: the role of the cytoplasmic ADP-glucose pyrophosphorylase. Plant Science 167: 1323-1333 Satagopan, S., Spreitzer R.J. (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J. Biol. Chem. 279: 14240-14244. Schiøtt, M., S. Romanowsky, M.K. Jakobsen, L. Bækgaard, M.G. Palmgren, J.F. Harper (2004) A plant plasma membrane Ca2+ pump is required for normal pollen tube growth and fertilization. PNAS 101: 9502-9507. Sebastia, C.H., S.C Hardin, S.D.Clouse, J.J. Kieber, S.C. Huber (2004) Identification of a new motif for CDPK phosphorylation in vitro that suggests ACC synthase may be a CDPK substrate. Arch. Biochem. Biophys. 428: 81-91. Sebastia, C.H., F. Marsolais, C. Saravitz, D.W. Israel, R. Dewey, S.C. Huber (200X) Possible role of amino acids in the control of storage product accumulation in developing soybean seeds. J. Exp. Botany, submitted. Seebauer JR, SP Moose, BJ Fabbri, LD Crossland, FE Below (2004) Amino acid metabolism in maize earshoots: Implications for assimilate preconditioning and nitrogen signaling. Plant Physiol. (in press). Stessman, D., M. Spalding, S. Rodermel (2004) Short and long term regulation of photosynthesis during leaf development. In Handbook of Photosynthesis. M. Pessarakli, ed. (Marcel Dekker, Inc., New York) (in press). Tovar-Mendez, A., Hirani T.A., Miernyk J.A., Randall D.D. (2004). Analysis of the catalytic mechanism of pyruvate dehydrogenase kinase. Arch. Biochem. Biophys. In press. Trievel, R.C., E.M. Flynn, R.L. Houtz, J.H. Hurley (2003) Mechanism of multiple lysine methylation by the SET domain enzyme Rubisco LSMT. Nature Structural Biology 10: 545-552. Uribelarrea, M, FE Below, SP Moose (2004) Grain composition and productivity of maize hybrids derived from the Illinois protein strains in response to variable nitrogen supply. Crop Sci. 44:1593-1600. Voznesenskaya, EV, S Choung, GE Edwards, VR Franceschi (200X) Distribution of components of C4 photosynthesis in the triple layered chlorenchyma of Aristida (Poaceae). Functional Plant Biology, In press. Voznesenskaya EV, S Chuong, O Kiirats, VR Franceschi, GE Edwards (200X) Evidence that C4 species in genus Stipagrostis, family Poaceae, is NADP-malic enzyme subtype with nonclassical type of Kranz anatomy (Stipagrostoid). Plant Science. In press. Voznesenskaya EV, VR Francheschi, GE Edwards (2004) Light dependent development of single cell C4 photosynthesis in cotyledons of Borszczowia aralocaspica (Chenopodiaceae) during transformation from a storage to a photosynthetic organ. Annals of Botany 93: 1-11. Wachter, R., M. Langhans, R. Aloni, S. Gotz, A. Weilmunster, A. Koops, L. Temguia, I. Mistrik, J. Pavlovkin, U. Rascher, K. Schwalm, K. E. Koch, C. I. Ulrich (2003) Vascularization, high-volume solution flow, and localized roles for enzymes of sucrose metabolism during tumorigenesis by Agrobacterium tumefaciens. Plant Physiology. 133:1024-1037. Walker D, G Edwards (2004) Photosynthetic Carbon Assimilation. In: Molecular to Global Photosynthesis. Series on Photoconversion of Solar Energy, v. 2. Eds. MD Archer and J Barber. pp. 189-220. Invited chapter. Singapore: World Scientific Pres. Weltzin, J.F. M.E. Loik, S. Schwinning, D.G. Williams, P. Fay, B. Haddad, J. Harte, T.E. Huxman, A.K. Knapp, G. Lin, W.T. Pockman, M.R. Shaw, E. Small, M.D. Smith, D.T. Tissue, J.C. Zak (2003) Assessing the response of terrestrial ecosystems to potential changes in precipitation. BioScience 53: 941-952. Wubben, M.J.E., S.R. Rodermel, T.J. Baum (2004) Mutation of a UDP-glucose-4-epimerase alters root ethylene responses and nematode susceptibility in Arabidopsis thaliana. Plant Journal (in press). Yep, A., M. A. Ballicora, M. N. Sivak, J. Preiss (2004) Identification and characterization of a critical region in the glycogen synthase from Escherichia coli. J. Biol. Chem. 279: 8359-8367. Yep, A., M. A. Ballicora, M. N. Sivak, J. Preiss (2004) The active site of the Escherichia coli glycogen synthase is similar to the active site of retaining GT-B glycosyltransferases. (2004) Biochem. Biophys. Res. Commun. 316: 960-966. Yep, A., C.M. Bejar, M.A. Ballicora, J.Dubay, A.A. Iglesias, J. Preiss, (2004) An assay for the ADP-glucose pyrophosphorylase that measures the synthesis of radioactive ADP-glucose with glycogen synthase. Analytical Biochemistry. 324: 52-59. Yu, F., S. Park, S.R. Rodermel (2004) The Arabidopsis FtsH metalloprotease gene family: interchangeability of subunits in chloroplast oligomeric complexes. Plant Journal 37: 864-876. Zheng, P., K. Ammar, A.-M. Girard, S. Rodermel, D.R. Thomas, L. Ning, J.B. Callis, G.E. Edwards, L. Daley (2003) Test of an in vivo method to detect chloroplast division in crop plants. Part III: Statistical proofs of observation and general utility of the method. Spectroscopy 18: 102-105. ZHU X, PORTIS AR, LONG SP (2004) Would transformation of C3 plants with foreign Rubisco increase productivity? Plant Cell Environment 27: 155-165. ZHU X.G., ORT D.R., WHITMARSH J., LONG S.P. (2004) The slow reversibility of photosystem II thermal energy dissipation on transfer from high to low light may cause large losses in carbon gain by crop canopies: a theoretical analysis. J Exp Botany 55: 1167-1175. NC-1-142 PATENT APPLICATION: H. Winter, S.C. Huber, C. Larabell (Filed Oct 17, 2004) (LBNL Docket No. JIB-1571) Synthetic peptides that cause F-actin bundling and block actin depolymerization. NC-1-142 PATENTS AWARDED: Randall, D.D., Mooney B.P., Johnston M.L, Luethy M.H., Miernyk J.A. (2004) U.S. Patent No. 6,773,917 entitlted: Use of DNA encoding plastid pyruvate dehydrogenase and branched-chain oxoacid dehydrogenase components to enhance polyhydroxyalkanoate biosynthesis in Plants.