Annual/Termination Reports:

[09/23/2008] [12/07/2009] [12/15/2009] [01/12/2011] [11/30/2011] [06/12/2013]

Report Information

Participants

Fred Below (University of Illinois), Christoph Benning (Michigan State University), Hans Bohnert (University of Illinois), Raymond Chollet* (University of Nebraska), John Cushman (University of Nevada), Gerald Edwards (Washington State University), Glenda E. Gillaspy (Virginia Tech), Irwin Goldman (University of Wisconsin, Administrative Advisor), Mark Guiltinan (Pennsylvania State University), Jeff Harper (University of Nevada), Robert Houtz (University of Kentucky), Steve Huber (USDA-ARS, University of Illinois), Jyan-Chyun Jang (Ohio State University), Robert Jones (University of Minnesota), Karen Koch (University of Florida), Wayne Loescher (Michigan State University), Stephen Long (University of Illinois), John Markwell* (University of Nebraska), Gail McLean (USDA, CSREES Representative), Ron Mittler (University of Nevada), Brandon Moore (Clemson University), Thomas Okita (Washington State University), Archie Portis* (USDA-ARS, University of Illinois), Jack Preiss (Michigan State University), Steven Rodermel (Iowa State University), Mary E. Rumpho (University of Maine), Michael E. Salvucci (USDA-ARS, Maricopa, AZ), Martin Spalding (Iowa State University), Robert Spreitzer (University of Nebraska), Donald P. Weeks (University of Nebraska) [*retired, last meeting]

Brief Summary of Minutes

This was the first meeting of Multistate Research Project NC1168 (Regulation of Photosynthetic Processes), which is the approved renewal of NC1142. The meeting was hosted by the Nebraska AES and held at the Beadle Center of the University of NebraskaLincoln on November 17, 2007.

Attending members included Christoph Benning, Raymond Chollet, Gerald Edwards, Glenda Gillaspy, Irwin Goldman, Karen Koch, Wayne Loescher, John Markwell, Gail McLean, Archie Portis, Jack Preiss, Mary Rumpho, Michael E. Salvucci, Robert J. Spreitzer, Donald P. Weeks, and Marna Yandeau-Nelson (for Guiltinan). Attending guests included Gary Cunningham (Dean and Director of the Nebraska AES) and Julie Stone (University of Nebraska).

The meeting convened at 8:50 AM in room E228 in the Beadle Center. Bob Spreitzer (Nebraska, AES) provided opening remarks and introduced Dr. Gary Cunningham, Dean and Director, Nebraska AES.


Bob Spreitzer then gave the first presentation on Rubisco engineering studies. A new member, Mary Rumpho, Maine AES, then presented her work on a sea slug that engulfs and maintains chloroplasts. Bob Spreitzer then introduced Irwin Goldman, NC1168 project administrator, who drove in from Madison, WI. Archie Portis, Illinois ARS, presented his work on genetic engineering of Rubisco in plants and member Hans Bohnerts work on a new algorithm to correlate gene expression patterns with gene function. At 10:35 AM a break occurred, and, at 10:55, Mike Salvucci, Arizona ARS, spoke about next years meeting in Arizona, an upcoming Gordon Conference on photosynthesis, and his work on Rubisco activase and heat stress. Don Weeks, Nebraska AES, spoke about carbon-concentrating mechanisms, a collaboration with member Martin Spalding. Lunch occurred in the atrium of the Beadle Center at 11:35 (hosted by the director of the Nebraska AES). The meeting reconvened at 1:00.

Gerry Edwards, Washington AES, described his work and also member Tom Okitas work on the AGPase enzyme. Karen Koch, Florida AES, spoke about sorbitol and carbon partitioning, followed by Marna Yandeau-Nelson, who spoke for member Mark Guiltinan, Pennsylvania AES, on the starch branching enzymes. After a break, Jack Preiss, Michigan AES, described his structural work, and added that this is perhaps his last meeting. Wayne Loescher, Michigan AES, spoke about sugar alcohols and plant stress. After a break, Glenda Gillaspy, Virginia AES, followed with a talk on sugar-alcohol synthesis genes. Christoph Benning, Michigan AES, spoke about plans to produce oils in canola that can be used for biodiesel production.

After a break, Irwin Goldman, Administrative Advisor, Wisconsin AES, provided an update on the last project (NC-1142), relaying that Bob Spreitzer was key in getting the new project (NC-1168) approved. Irwin then outlined the process of NCRA and encouraged members to let their Experiment Station directors know about NC1168 activities and the importance of this project. Irwin described how he could help organize an NC1168 meeting in the future and could provide assistance to the members of NC-1168. Irwin also announced that a progress report for NC1142 would be due soon, and that he would send inquires out by email at the beginning of the year requesting information for this report. Gail McLean, USDA/CSREES Representative then presented an update on USDA/NRI programs relevant to members. She outlined the current funding level and outlook for several programs. She added that it is important for this group to file annual reports.

At 5:05 PM the business meeting convened. Bob Spreitzer brought up the next and future meeting dates as related to whether the group should stay with the second weekend in November or switch to the third. Mike Salvucci remarked that hotel rooms in Arizona would be cheaper earlier in the season. After a discussion, the group decided to retain the second weekend in November as the meeting time for future meetings. Bob Spreitzer then remarked on the status of attending and non-attending members, adding that all members are active in attending the meetings. Jack Preiss inquired about Doug Randall, and Bob Spreitzer replied that Randall had retired. Bob Spreitzer announced a need for a new secretary, who would host the meeting in 2011, and who would be in charge of the project renewal. Christoph Benning was nominated and then elected to serve in this position. The renewal will be due in the Spring of 2012.

The schedule for future meetings was determined:

2008 Arizona AES Mike Salvucci
2009 Ohio AES J. C. Jang
2010 Virginia AES Glenda Gillaspy
2011 Michigan AES Christoph Benning

The meeting was adjourned at 5:20 PM.

Accomplishments

A. Plastid Function and Intracellular Communication. <br /> <br /> IA-AES has continued to investigate the role of the var2 gene of Arabidopsis (which encodes a chloroplast AtFtsH metalloprotease) in the repair of photodamaged photosystem II. Selection of suppressors has now identified genes encoding a polyribonucleotide phosphorylase and a chaperonin 60a that may also contribute to the assembly and regulation of the photosynthetic apparatus.<br /> <br /> ME-AES has continued to study the endosymbiotic relationship between the marine mollusk Elysia chlorotica (sea slug) and chloroplasts of the heterokont alga Vaucheria litorea. Sequences for phosphoribulokinase and the photosystem II Mn-stabilizing protein have been found in sea slug DNA, indicating that horizontal gene transfer may have occurred as a means for maintaining photosynthesis in the endosymbiotic algal chloroplasts.<br /> <br /> B. Photosynthetic Capture and Photorespiratory Release of CO2. <br /> <br /> IA-AES has identified a gene (lciB) responsible for the CO2-concentrating mechanism in the green algae Chlamydomonas reinhardtii. Further analysis, in collaboration with NE-AES, will evaluate the potential for transferring all or part of the CO2-concentrating mechanism into crop plants. NE-AES is using RNAi to study a possible bicarbonate transporter that may be involved in the CO2-concentrating mechanism.<br /> <br /> WA-AES is studying unique species in the family Chenopodiaceae that perform C4 photosynthesis without Kranz anatomy. Microtubules are critical for the maintenance of two cytoplasmic compartments. One type of chloroplast has Rubisco, starch biosynthesis, and well-developed grana, while the other type of chloroplast is specialized for supporting CO2 fixation in the C4 cycle. <br /> <br /> AZ-ARS and IL-ARS have continued to collaborate on Rubisco activase. An S-tagged b-isoform of activase has been engineered in Arabidopsis and crossed with an Arabidopsis plant expressing only the a-isoform. The new line will be used to examine the effects of temperature on interactions among activase subunits, and the effect of these interactions on Rubisco regulation. IL-ARS has transformed more-thermostable Rubisco activase into Arabidopsis to demonstrate improved photosynthesis and growth at elevated temperature.<br /> <br /> IL-ARS is attempting to eliminate rbcS gene expression in tobacco by employing RNAi technology. The Amaranthus rbcS gene has been introduced as a first step to see whether C4 Rubisco may provide a benefit for CO2 fixation. NE-AES has produced hybrid Rubisco enzymes in Chlamydomonas comprised of crop-plant small subunits and Chlamydomonas large subunits. Small subunits influence large-subunit catalysis. Phylogenetic analysis and directed mutagenesis have shown that large-subunit regions far from the active site can also influence catalysis.<br /> <br /> KY-AES has determined the polypeptide substrate specificity determinants for pea Rubisco large subunit methyltransferase using a fusion protein construct between the first 23 amino acids of the large subunit of Rubisco and human carbonic anhydrase II. RNAi-mediated knockdown of the methyltransferase in transgenic tobacco resulted in a two-fold decrease of a-tocopherol, the product of g-tocopherol methyltransferase, indicating that its consensus sequence may also be methylated. The crystal structure of Arabidopsis peptide deformylase 2 has been determined, and its substrate specificity is being defined.<br /> <br /> C. Mechanisms Regulating Photosynthate Partitioning. <br /> <br /> FL-AES has investigated a role for sorbitol metabolism and/or shuttling within developing kernels, possibly involving dual contributions to carbon transport and modulation of reducing power. Analysis of maize mutants has shown that sorbitol metabolism can contribute prominently to kernel capacity for sucrose import and use. FL-AES has also developed a method for 3'-UTR profiling that resolves expression to the level of individual gene family members and facilitates gene discovery. <br /> <br /> By whole-genome analysis, IL-AES has discovered that genes involved in nitrogen sensing and signaling may limit productivity in crop plants grown under elevated CO2 conditions. NV-AES has been identifying targets in nitrogen and carbon metabolism that interact with affinity-tagged 14-3-3 regulatory proteins in Arabidopsis. <br /> <br /> OH-AES is studying bZIP transcription factors involved in sugar signaling. SC-AES has shown that F-actin mediates some aspects of glucose signaling in Arabidopsis by interacting with mitochondrial hexokinase 1. Directed mutagenesis indicates that the hexokinase-like proteins, which lack glucose phosphorylation activity, may have specialized roles as regulatory components for glucose signaling. VA-AES is studying the synthesis and signaling functions of the sugar alcohol myo-inositol, which is linked to osmoprotective functions, signal transduction, and ascorbic acid synthesis, among other functions. Arabidopsis mutants lacking various members of the myo-inositol phosphate synthase and inositol monophosphatase enzyme families indicate that there are different subcellular locations for the first and second steps in myo-inositol synthesis.<br /> <br /> MI-AES is investigating the function and molecular biology of WRINKLED 1 and the genes controlled by this transcription factor in Arabidopsis. Analysis of mutants and engineered genes are clarifying the role of WRINKLED 1, pyruvate kinase, and glucose-6-phosphate dehydrogenase in the partitioning of photosynthetic carbon to oils. Understanding the conversion of photosynthetic sugars into triacylglycerols will be essential for the engineering of novel biofuel crops.<br /> <br /> MI-AES and WA-AES continue to define the structure-function relationships of ADP-glucose pyrophosphorylase, which catalyzes the first step in starch synthesis. Families of genes encoding the subunits have been analyzed in Arabidopsis. Hybrid enzymes and directed mutagenesis are being used to define regions involved in catalysis and regulation. This information will be used for the further engineering of improvements in the production of starch. WA-AES has used up-regulated forms of ADP-glucose pyrophosphorylase to increase leaf starch and biomass of Arabidopsis and rice. PA-AES is studying the starch branching enzymes of maize, which are critical in determining the molecular fine-structure of starch. Recent data indicate that starch branching enzyme activity in leaf extracts can be modulated by redox reactions. <br /> <br /> D. Developmental and Environmental Limitations to Photosynthesis. <br /> <br /> AZ-ARS and IL-ARS have shown that inhibition of photosynthesis and loss of Rubisco activation under moderate heat stress is more acute when Rubisco activase levels are suboptimal for photosynthesis, regardless of the isoform of activase. This finding is consistent with a proposed mechanism for inhibition of photosynthesis by heat stress based on the acute sensitivity of activase to thermal denaturation. New data indicate that chaperonin-60b plays a role in acclimating photosynthesis to heat stress, possibly by protecting Rubisco activase from thermal denaturation.<br /> <br /> IL-AES has found that chemical treatments promoting leaf greening in maize can disrupt ear development by altering ethylene sensing.<br /> <br /> IL-AES continues to exploit soyFACE (http://www.soyface.uiuc.edu) for investigating the effects of elevated CO2 on plant productivity. Metabolic profiling and gene-mining methods are being developed to analyze environmental stress effects. Gene networks are being defined.<br /> <br /> MI-AES and WA-AES have shown that expression of the mannitol biosynthetic enzyme gene, M6PR, in Arabidopsis improves salt tolerance. MI-AES is also studying the constitutive expression of the abiotic stress responsive transcription-factor gene, CBF3, and the plasma-membrane sodium antiporter gene, SOS1, in transgenic Arabidopsis to see whether further improvements in salt tolerance can be achieved.<br /> <br /> VA-AES is studying the role of myo-inositol signaling in abiotic stress. Protein-protein interactions in sugar signaling pathways are being identified.<br /> <br /> NV-AES has used microarray transcript profiling, quantitative RT-PCR, and metabolite profiling to show that metabolite differences found under long-term salinity or water deficit stress are linked to differences in transcript abundance of many genes involved in energy metabolism and nitrogen assimilation, particularly photosynthesis, gluconeogenesis, and photorespiration. Water-deficit-treated plants also appeared to have a higher demand than salinized plants to adjust osmotically, detoxify free radicals, and cope with photoinhibition. NV-AES is also screening through Arabidopsis mutants to identify genes that affect a plant's ability to grow under extreme environments. These genes provide important leads towards a goal of engineering higher-yield crops. <br />

Publications

Impact Statements

1. Further study of a protease, polyribonucleotide phosphorylase, and chaperonin 60a that play a role in the repair of photodamaged photosystem II may lead to the design of strategies for manipulating the photosynthetic capacity and quality of crop plants.
2. A deeper understanding of the endosymbiotic association between sea slug and algal chloroplasts may enable the maintenance of isolated organelles in culture or foreign hosts for long periods of time, thereby enabling large-scale production of chloroplast secondary products and/or alternative forms of photosynthetic energy production.
3. Transfer of the algal CO2-concentrating mechanism or C4 photosynthesis to C3 crop plants may improve photosynthetic productivity.
4. The discovery of a possible interaction between Rubisco activase and chaperonin-60 provides a new avenue for research into photosynthetic acclimation to moderate heat stress.
5. Improved thermal resistance of photosynthesis has been achieved by engineering a more-thermostable Rubisco activase.
6. Regions far from the active site have been identified as potential targets for the improvement of Rubisco and photosynthetic productivity.
7. Posttranslational methylation may be involved in the regulation of several chloroplast-localized enzymes. Further study may increase our understanding of how carbon reactions in the chloroplast can be altered to enhance photosynthetic efficiency.
8. The sdh1 (sorbitol dehydrogenase 1) maize mutant offers us a means for testing newly-revealed, but little-known roles of sugar alcohols in grain development.
9. Hexokinase-like proteins are not merely compromised in catalytic activity, but have specialized non-catalytic functions important for sugar signaling.
10. An alternate pathway for the synthesis of ascorbic acid from myo-inositol has been discovered. Because ascorbic acid is both an essential plant-derived nutrient for humans, and an important anti-oxidant in plants, understanding its synthesis can lead to new strategies for improving both human and plant health.
11. By developing schemes and new biofuel crops that lead to improved oil yield per acre of five-fold or more, biodiesel from plant oils could make up a significant fraction of renewable fuels.
12. ADP-glucose pyrophosphorylase has been shown to be a useful target for engineering increases in the production of starch and biomass.
13. New knowledge about the mechanisms of starch branch-point formation and its role in starch granule formation will lead to the development of novel starch types for industrial, medical, and food-processing applications.
14. Knowing when to apply chemical treatments to achieve leaf-greening without disrupting maize-ear development is clearly important to the agrochemical industry.
15. Peptide deformylase provides a unique opportunity for exploring potential alternatives to antibiotic-resistance markers in plant transformation vectors as well as alternative broad-spectrum herbicides.
16. In soyFACE elevated-CO2 experiments, superior ecotypes have been identified that are less affected by changes in environmental stress parameters.
17. Genetic engineering of improved salt tolerance has been achieved in Arabidopsis without a decline in photosynthesis.
18. Calcium signaling pathways are being defined that are responsible for plant survival under extreme environmental conditions such as cold, heat, drought, salt, and high-light stress.

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Report Information

Participants

Brief Summary of Minutes

Please see attached minutes file for annual report.

Accomplishments

Publications

Impact Statements

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Report Information

Participants

Brief Summary of Minutes

Please see attached meeting minutes file for annual report.

Accomplishments

Publications

Impact Statements

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Report Information

Participants

Robert Aiken (Kansas State University), Fred Below (University of Illinois), Christoph Benning (Michigan State University), Hans Bohnert (University of Illinois), John Cushman (University of Nevada), Gerald Edwards (Washington State University), Glenda Gillaspy (Virginia Tech), Irwin Goldman (University of Wisconsin, Administrative Advisor), Mark Guiltinan (Pennsylvania State University), Jeff Harper (University of Nevada), Robert Houtz (University of Kentucky), Steve Huber (USDA-ARS, University of Illinois), Jyan-Chyun Jang (Ohio State University), Robert Jones (University of Minnesota), Karen Koch (University of Florida), Shing Kwok (USDA, CSREES Representative), Jiaxu Li (Mississippi State University), Wayne Loescher (Michigan State University), Stephen Long (University of Illinois), Ron Mittler (University of Nevada), Brandon Moore (Clemson University), Thomas Okita (Washington State University), Vara Prasad (Kansas State University), Steven Rodermel (Iowa State University), Mary E. Rumpho (University of Maine), Michael E. Salvucci (USDA-ARS, Arizona), Martin Spalding (Iowa State University), Robert Spreitzer (University of Nebraska), Donald Weeks (University of Nebraska)

Brief Summary of Minutes

The annual meeting of Multistate Research Project NC1168 (Regulation of Photosynthetic Processes) was hosted by the Virginia AES and held at the Peggy Lee Hahn Garden Pavilion at Virginia Tech, Blacksburg, Virginia on November 13, 2010.

Attending members included Robert Aiken, Christoph Benning, Gerald Edwards, Glenda Gillaspy, Jiaxu Li, Jeff Harper, Michael E. Salvucci, Robert J. Spreitzer, and Donald P. Weeks. Attending guests included Saied Mostaghimi (Associate Dean and Director of the Virginia AES), Eva Collakova (Virginia Tech), and Jim Moroney (Louisiana State University).

The meeting convened at 9:00 AM at the Hahn Garden Pavilion. Glenda Gillaspy (Virginia, AES) introduced Dr. Saied Mostaghimi, Associate Dean and Director, who summarized the organization and progress of the Virginia AES. Mike Salvucci (Arizona ARS) gave the first talk pertaining to objective 2. He described the structural basis for interactions between Rubisco and Rubisco activase (in collaboration with the Nebraska AES), and described field studies aimed at elucidating the influence of Rubisco activase on thermal tolerance. Bob Spreitzer's presentation focused on Rubisco engineering, including collaborative results from the Arizona ARS pertaining to the Rubisco structural transition. Gerry Edwards (Washington AES) described recent progress on mechanisms of single-cell C4 photosynthesis. After a short break (11:25-11:45 AM), the meeting resumed with Don Weeks summarizing recent collaborative work (with Iowa AES) in the application of TAL effector nucleases. Lunch was held at the Inn at Virginia Tech at 12:25 PM. The group returned to the Hahn Pavilion at 1:40 PM, at which time Jim Moroney (a potential new member from the Louisiana AES) brought the group up to date with regard to his research on the algal CO2 concentrating mechanism. He focused his presentation on a protein (LCI1) involved in carbon acquisition, a new protein (CIA6) that may play a role in pyrenoid formation, and various carbonic anhydrases. Christoph Benning (Michigan AES) gave the first talk pertaining to objective 3. He described progress in the use of rutabaga and algae for oil production. Eva Collakova (guest from Virginia Tech) discussed the utility of metabolic flux analysis with regard to metabolic engineering. After a short break (3:30-3:50 PM), Jeff Harper (Nevada AES) and Glenda Gillaspy (Virginia AES) described research at the interface between objectives 3 and 4. Harper discussed calcium signaling, and Gillaspy presented results on the role of inositol phosphates in carbon partitioning and signaling. In objective 4, Jiaxu Li (Mississippi AES) described histone methyltransferases involved in gene regulation. Rob Aiken (Kansas AES) described field studies focused on drought and oxidative stress.

After a 15 minute break, the business meeting was called to order by the chair (Gillaspy) at 6:00 PM. Irwin Goldman (Administrative Advisor) joined the group via Skype. Goldman stressed the importance of communicating the integration and collaboration of group members in future annual reports. There was confusion about who serves as secretary for the group. It was decided/confirmed that the chair-elect is responsible for taking minutes at the annual meeting. Spreitzer voiced concern about attendance of members at the annual meeting. Because of the broad, geographical distribution of participating members, travel to the annual meeting may be difficult. Harper proposed that the group meet at a more-accessible "hub" city. After discussion, it was concluded that the host of the annual meeting would choose the location. However, Salvucci offered to serve as chair of the meeting in Phoenix (in 2013) as a more-accessible and previously-favored location. The group then considered the election of Jim Moroney as a member of NC1168. The vote of members present, and those communicating via email, was unanimous. The group discussed the renewal application of NC1168. It was agreed that the four objectives will remain the same. Benning will take the lead in preparing the application. Benning will coordinate members' input to objective 1. Spreitzer will take the lead on objective 2. Gillaspy will coordinate objective 3. Salvucci will organize members in objective 4. Goldman will provide information on important dates/deadlines for preparation of the renewal proposal.

The appointment of chairs and schedule for future meetings are:
2011 Michigan AES Christoph Benning
2012 Maine AES Mary Rumpho (secretary, chair-elect in 2011)
2013 Arizona ARS Mike Salvucci (secretary, chair-elect in 2012)

The meeting was adjourned at 6:45 PM. Members met as a group for dinner on both November 12 and 13 for discussions of research results and for establishing collaborative plans.

Accomplishments

Obj. A. Plastid Function and Intracellular Communication. Photosynthesis is comprised of tightly coupled reactions requiring finely-tuned nucleocytosolic-plastid interactions and carefully executed signaling pathways. Efforts in this area center on the examination of a unique plastid model system (Maine AES) and studies of sugar signaling pathways (Ohio and Virginia AES). ME-AES continued characterization of expression of the photosynthetic mollusc, Elysia chlorotica, which harbors plastids intracellularly obtained from the algae, Vaucheria litorea. Quantitative PCR was used to compare transcript levels of both nuclear (psbO and prk) and plastid (psaA, psbA and rbcL) photosynthesis genes in E. chlorotica and V. litorea. In all cases, transcript levels were lower in the sea slug than its algal prey and they did not respond similarly to changes in light conditions over a 24-hr period. In collaboration with NC-1168 PI Wayne Loescher (MI AES), work has begun to quantify the major photosynthetic products of both organisms. Mannitol, hexoses and cyclitols have been found.<br /> <br /> Work on sugar signaling pathways seeks to understand the molecules that plants use in these pathways. OH-AES characterized the effects of a group of bZIP transcription factors and a tandem zinc finger (TZF) gene family in sugar-mediated transcriptional and post-transcriptional regulation, respectively. They have found that the expression of AtbZIP1 is repressed by sugars in a fast, sensitive, and reversible way. This sugar repression is also affected by a conserved sugar signaling component, hexokinase. VA-AES has investigated an energy sensor called SnRK1.1 which plants use to sense sugars and energy. It was found that SnRK1.1 is regulated by proteasomal turnover and proteins have been identified that participate in this process.<br /> <br /> Obj. B. Photosynthetic Capture and Photorespiratory Release of CO2. Rubisco catalyzes the rate-determining step in photosynthesis, but it does so very inefficiently. Because of this inefficiency, Rubisco presents an obvious target for increasing photosynthetic performance. The Arizona ARS and Nebraska AES have been working to understand key factors in Rubisco activity. Because the catalytic properties of Rubisco enzymes differ among various species, NE-AES has been taking a phylogenetic approach to identify those regions of the Rubisco large subunit that may account for these differences. Chlamydomonas mutant enzymes have been characterized with decreased CO2/O2 specificity, but it can be activated normally by Rubisco activase, indicating that the structural transition required for catalysis is not affected. It appears that there may be a different path to the active site that is influenced by the nature of the large/small-subunit interface. In synergistic studies, AZ-ARS has sought to understand Rubisco activase structure/function by acquiring structural information at the atomic level. In collaboration with NE-AES they have examined the effect of substituting higher plant small subunits (RBCS) on the activation of Chlamydomonas Rubisco by activase. This finding suggests that replacing native RBCS in Chlamydomonas Rubisco with a higher plant RBCS increases the stability of the binary Rubisco-activase complex without affecting the rate of Rubisco activation.<br /> <br /> Two members from the Washington AES have focused on research on the structural and biochemical diversity in mechanisms of C4 photosynthesis which evolved to concentrate CO2 around Rubisco, enhance photosynthesis, and reduce losses of CO2 due to photorespiration. These members reviewed current information on structural and biochemical diversity in C4 photosynthesis and described 25 known structural forms of Kranz anatomy and two structural forms where C4 occurs within individual photosynthetic cells without Kranz. Other efforts focused on the characterization of the mechanism of C4 in single-cell C4 species in which photosynthesis occur within individual photosynthetic cells (Chenopodiaceae). Two cytoplasmic domains develop with dimorphic chloroplasts considered to function analogous to mesophyll and bundle sheath cells in Kranz type species. WA-AES is working on the biochemical, physiological properties and molecular requirements for this form of C4. A purification protocol for the two different chloroplast types has been developed, and functional studies performed on intact chloroplasts. The results provide evidence that one domain is specialized for supporting fixation of atmospheric CO2 in the C4 cycle, and the other for accepting CO2 from decarboxylation of C4 acids and its assimilation by Rubisco in the C3 cycle.<br /> <br /> Obj. C. Mechanisms Regulating Photosynthate Partitioning. Several approaches are underway to understand how plants regulate photosynthate partitioning, and this area of work has important implications for generating better bioenergy crops. Since increasing oil biosynthesis in vegetative tissues of plants offers a way to convert carbon and increase the energy density of biomass, Michigan AES has examined converting starch to oil in the rutabaga storage organ. A transcriptomic analysis in different rutabaga tissues is underway to identify hypocotyl specific promoters and new target genes for second generation transgenics. MI-AES is also exploring the induction of triacylglycerol biosynthesis in microalgae. Synergistic studies are underway at the Nevada AES focusing on the algae Dunaliella salina. This unicellular, halophytic green alga belongs to the Chlorophyceae, and is among the most industrially important microalgae because it can produce massive amounts of ²-carotene and because of its potential as a feedstock for biofuels production. A complementary DNA (cDNA) library was constructed from D. salina cells adapted to 2.5 M NaCl and used to generate an expressed sequence tag (EST) database. ESTs were obtained for 2,831 clones representing 1,401 unique transcripts. Putative functions were assigned to 1,901 (67.2%) ESTs after comparison with protein databases. An additional 154 (5.4%) ESTs had significant similarity to known sequences whose functions are unclear and 776 (27.4%) had no similarity to known sequences. A third synergistic effort is underway by two members (Nebraska AES and Iowa AES) to develop a transformation system for algae. The ability to readily disrupt genes and/or replace gene sequences using homologous recombination has greatly speeded research with bacterial and yeast systems. When developed, this is expected to dramatically facilitate studies in algae, plants, and development of bioenergy resources. These NC-1168 members have tested a TAL effector nuclease (TALN) technology for targeted gene knockout and gene replacement in algae. To test their gene-specific TALN technology, preliminary experiments have been conducted in which naturally-occurring full-length TAL effectors were tested for the specificity of their binding to target DNA both in vitro and in vivo. The results suggest that artificial TALNs can be produced to target specific genes. <br /> <br /> A group of NC-1168 members have examined signaling components that impact photosynthate partitioning (Illinois-ARS, Nevada AES, Virginia AES). IL-ARS is examining engineering photosynthesis by a directed mutagenesis of a receptor kinase. It was found that a receptor kinase, BAK1, needs to be phosphorylated on a tyrosine residue in vivo for this. Preventing autophosphorylation by directed mutagenesis enhances plant growth. The basis for increased growth may be a higher rate of light saturated CO2 assimilation under ambient conditions, and accordingly these plants accumulate higher levels of starch and sucrose in leaves during the day. Transcriptome analysis indicated that the expression of numerous genes was altered and this may suggest new approaches to indirectly increase whole plant carbon assimilation. NV-AES has examined 14-3-3s, which are regulatory proteins that have been implicated in nitrogen and carbon metabolism by the NV-ARS. More than 124 14-3-3 clients have been identified, 103 of which have not previously been reported. Many of the newly identified clients are involved directly in metabolism, such as phosphoenol pyruvate (PEP) carboxylase. The VA-AES has investigated myo-inositol synthesis and phosphoinositide signaling molecules that appear to limit plant size. Phosphatidylinositol was found to also limit signaling by another important class of lipid signaling molecules, the sphingolipids. <br /> Three members are investigating metabolic controls on photosynthate partitioning. The Washington AES has examined the plastidic phosphorylase. The plastidic starch phosphorylase, Pho1, is essential for starch initiation at low temperatures. Although previous kinetic studies suggested that Pho1 was not involved in starch synthesis but rather starch degradation, kinetic analysis of the purified recombinant Pho1 indicates that it strongly favors the synthesis reaction. This may aid efforts to understand and modify source-sink relationships to improve crop productivity. Florida AES, and Michigan AES reported data on polyols in plants, which can be major sinks for carbon. FL-AES has found that sorbitol dehydrogenase has a role in determining the kernel number per ear in corn. In addition, the Cellulose Synthase-Like D (CSLD) genes were genetically determined to impact plant cell size. Key to this work are contributions to the UniformMu maize project which serves as a resource for sequence-indexed insertional mutants. The UniformMu population provides both seeds and sequences free of charge through the MaizeGDB. As of December 2010, 22,239 unique, germinal Mu transposon insertions have been mapped. FL-ARS and MI-AES are collaborating on the role of mannose, characterizing the impact of M6PR gene both in Arabidopsis and its original source, celery.<br /> <br /> Obj. D. Developmental and Environmental Limitations to Photosynthesis. Several NC-1168 members have pursued strategies to understand how photosynthesis can be limited by stress. Work has focused on molecular, whole plant and field responses. AZ-AES has examined differences in thermal stability between wild type Rubisco activase and structurally-altered mutant recombinant proteins in vitro and when the proteins were expressed in transgenic Arabidopsis plants. Rubisco activation studies indicated that that the thermal stability of Rubisco activase determines the heat-sensitivity of photosynthesis. In synergistic work, Kansas AES has examined the possible relationship between expression levels of Rubisco activase and plant yield in winter wheat, corn and Arabidopsis following prolonged exposure to heat stress. A significant, positive, linear correlation was found between the expression of activase and plant productivity under heat-stress conditions. In significant pilot work, the relevance of Rubisco activase to the inhibition of photosynthesis by heat stress under field conditions in Arizona was investigated. The combined effects of heat and drought under field conditions for cotton cultivars that differed in their drought tolerance were measured as part of a larger study designed to develop the capacity for high-throughput phenotyping. These studies use multispectral data spanning visible, near infrared, and thermal infrared wavelengths, along with plant canopy geometric data, to detect and evaluate plant responses to heat and drought stress at a field site in Maricopa, Arizona. Other field studies conducted by Kansas-ARS have identified genotypes in sorghum, wheat and soybean with increased stress tolerance which could enhance primary productivity and grain yield formation. Other breeding efforts by MI-AES focused on common bean to develop a better understanding of physiological parameters, quantitative trait loci and genes associated with drought tolerance in this important food crop. <br /> <br /> To investigate molecular mechanisms of stress, the NV-AES has examined the function of CAM in reactive oxygen species (ROS) alleviation using a newly isolated CAM-deficient mutant of a facultative halophyte Mesembryanthemum crystallinum L. Salinity induced nocturnal malate synthesis in the leaves of wild-type plant, but not in the mutant. These results indicated that lower levels of ROS accompanied the performance of CAM, and that CAM might alleviate oxidative stress. Another member of NV-ARS has also focused on several genes implicated in both calcium signaling and survival under extreme environmental conditions, such as cold, heat, drought, salt, and high-light stress. The calcium-dependent protein kinases, cyclic nucleotide gated channels, and P-type ATPase ion pump genes have been studied via functional genomics approaches with mutant and transgenic plants. The vacuolar calcium pumps (ACA4 and ACA11) control calcium signals that can trigger programmed cell death and limit plant productivity. Mississippi ARS reported on another set of genes, the Arabidopsis SET domain genes. One of these (SDG8) is a histone methyltransferase responsible for di- and trimethylation of the lysine 36 residue of histone 3. MI-ARS showed that knockdown mutations of this methyltransferase markedly reduce the global level of H3 trimethylation in vivo and affect rice flower and grain development. These studies suggest H3K36 trimethylation plays a key role in reproductive organ development in rice. <br />

Publications

Aiken, R. and N.L. Klocke. 2010. Operational characteristics of sap flow heat gauges to quantify transpiration flux in corn. In Proceedings of the ASABE 5th National Decennial Irrigation Conference. Phoenix, AZ. Dec. 5-8, 2010.<br /> <br /> Alkayal MF, Albion RL, Tillett RL, Mark S. Lemos, Hernandez-Gomez L, Cushman JC. (2010) Expressed Sequence Tag (EST) profiling in salinity shocked Dunaliella salina reveals high expression of protein synthetic apparatus components. Plant Science. 179: 437-449. <br /> <br /> Barta C., Carmo-Silva A. E., and Salvucci, M.E. (2010) Purification of Rubisco activase from leaves or after expression in Escherichia coli. In R. Carpentier (ed.) Photosynthesis Research Protocols, Methods in Molecular Biology, vol 684. In press. <br /> <br /> Barta C., Carmo-Silva A. E., and Salvucci, M.E. (2010) Rubisco activase activity assays. In R. Carpentier (ed.) Photosynthesis Research Protocols, Methods in Molecular Biology, vol 684. In press. <br /> <br /> Barta, C., Dunkle, A.M., Wachter, R. M. and Salvucci, M.E. (2010) Structural changes associated with the acute thermal instability of Rubisco activase. Arch. Biochem. Biophys. 499: 17-25. <br /> <br /> Candace Myers, Shawn M. Romanowsky, Yoshimi D. Barron, Shilpi Garg, Corinn L. Azuse, Amy Curran, Ryan M. Davis, Jasmine Hatton, Alice Harmon, and Jeffrey F. Harper (2009) Calcium- Dependent Protein Kinases Regulate Polarized Tip-Growth in Pollen Tubes. The Plant Journal. 59:528-39. <br /> <br /> Carmo-Silva A. E., Barta C. and Salvucci, M.E. (2010) Isolation of ribulose-1,5-bisphosphate carboxylase/oxygenase from leaves. In R. Carpentier (ed.) Photosynthesis Research Protocols, Methods in Molecular Biology, vol 684. In press. <br /> <br /> Chan, R. Grumet, WH. Loescher Transcriptome analysis indicates that ABA pathway activation, redox quenching, and cell wall strengthening contribute to enhanced salt tolerance of mannitol-producing Arabidopsis Submitted .<br /> <br /> Chan, Z, Loescher, W, Grumet , R. Transcriptional variation in response to salt stress in Arabidopsis thaliana Submitted. <br /> <br /> Cushman JC, Oliver MJ, (2010) Understanding vegetative desiccation tolerance using integrated functional genomics approaches within a comparative evolutionary framework. In: Ecological Studies: Desiccation Tolerance in Plants. Eds: Ulrich Luttge, Erwin Beck, and Dorothea Bartels. Springer, Heidelberg. Chapter 16, pp. xxx-xxx. <br /> <br /> Djanaguiraman M, Prasad PVV, Seppanen M. 2010b. Selenium protects sorghum leaves from oxidative damage under high temperature stress by enhancing antioxidant defense system. Plant Physiology and Biochemistry 48: 999-1007. <br /> <br /> Djanaguiraman M, Prasad PVV. 2010. Ethylene production under high temperature stress causes premature leaf senescence in soybean. Functional Plant Biology 37: 1071-1084. <br /> <br /> Djanaguiraman M, Sheeba JA, Devi DD, Bangarusamy U, Prasad PVV. 2010a. Nitrophenolates spray can alter boll abscission rate in cotton through enhanced peroxidise activity and increased ascorbate and phenolics levels. Journal of Plant Physiology 37: 1-9. <br /> <br /> Donahue, JL, Alford, SR, Torabinejad, J, Kerwin, R, Nourbakhsh, A, Ray, WK, Lyons, B, Hein PP, and Gillaspy, GE. (2010) The Arabidopsis thaliana Myo- Inositol 1-Phosphate Synthase1 Gene Is Required for Maintenance of Myo- inositol Synthesis and Suppression of Cell Death. The Plant Cell 22: 888-903. <br /> <br /> Dong G, Ma DP, Li J. (2008). The histone methyltransferase SDG8 regulates shoot branching in Arabidopsis. Biochem. Biophys. Res. Commun. 373: 659-664 <br /> <br /> Dong G, Wang Z, Zou J, Peng Z, and Li J. A histone H3 lysine 36 methyltransferase is involved in regulation of flower and grain development in rice. Submitted for review. <br /> <br /> Edwards, GE and EV Voznesenskaya. 2011 C4 photosynthesis: Kranz Forms and Single-Cell C4 in Terrestrial Plants. A.S. Raghavendra and R.F. Sage (Eds.) 2011. C4 Photosynthesis and Related CO2 Concentrating Mechanisms. Advances in Photosynthesis and Respiration. Volume 32. Springer, Dordrecht, The Netherlands. pp. 29-61. <br /> <br /> Feodorova, TA, EV Voznesenskaya, GE Edwards and EH Roalson. Biogeographic patterns of diversification and the origins of C4 in Cleome (Cleomaceae). Systematic Botany, In press. <br /> <br /> Genkov T., Meyer, M., Griffiths, H., and Spreitzer, R. J. (2010) Functional hybrid Rubisco enzymes with plant small subunits and algal large subunits: Engineered rbcS cDNA for expression in Chlamydomonas. J. Biol. Chem. 285, 19833-19841 (Paper of the Week). <br /> <br /> Genkov, T., and Spreitzer, R. J. (2009) Highly conserved small subunit residues influence Rubisco large subunit catalysis. J. Biol. Chem. 284, 30105-30112. <br /> <br /> Gholipoor M, Prasad PVV, Mutava RN, Sinclair TR. 2010. Genetic variability of transpiration response to vapor pressure deficit among sorghum genotypes. Field Crops Research 119: 85-90. <br /> <br /> Gillaspy, GE (2010) The Polyphosphoinositide Phosphatases in Lipid Signaling in Plants. Springer; ed: T. Munnik. <br /> <br /> Gore, M.A., Coyle, G., Friebe, B., Coffelt, T.A. and Salvucci, M.E. (2010) Complex ploidy level variation in guayule breeding programs. Crop Sci., in press. <br /> <br /> Goren, S., Huber, S.C. and Granot, D. 201X. Evidence of developmental coordination of sucrose synthase isozymes in tomato plants. Planta, in revision. <br /> <br /> Ho, L-W, T-T Yang, S-S Shieh, GE Edwards, HE Yen. 2010 Reduced expression of a vesicle trafficking-related ATPase SKD1 decreases salt tolerance in Arabidopsis. Functional Plant Biol 37:962-973. <br /> <br /> Horan K, Jang C, Bailey-Serres J, Mittler R, Shelton C, Harper JF, Zhu JK, Cushman JC, Gollery M, Girke T. (2008) Annotating genes of known and unknown function by large-scale coexpression analysis. Plant Physiol. 147(1): 41-57. <br /> <br /> Hozain, M. I., Salvucci, M. E., Fokar, M., and Holaday, A. S. (2010) The differential response of photosynthesis to high temperature for a boreal and temperate Populus species relates to differences in Rubisco activation and Rubisco activase properties. Tree Physiol. 30: 32-44. <br /> <br /> Huber, S.C., Kaiser, W.M., and Jain, V. 201X. Posttranslational regulation of nitrate reductase. In: Nitrogen. Narosa Publishers, Delhi., in press. <br /> <br /> Hwang, S.-K., N. Crofts, H. Satoh, and T.W. Okita (2010) Biochemical evidence supports a role for the rice endosperm-specific plastidial !-glucan phosphorylase in starch synthesis. Archives Biochem. Biophys. 495:82-92. doi:10.1016/j.abb.2009.12.023<br /> <br /> Ing-Feng Chang, Amy Curran, Rebekah Woolsey, David Quilici, John Cushman, Ron Mittler, Alice Harmon, and Jeffrey Harper (2009) Proteomic profiling of tandem affinity purified 14-3-3 protein complexes in Arabidopsis thaliana. Proteomics. 9:2967-85. <br /> <br /> Kang, S.G., Price, J., Lin, P.C., Hong, J.C., and Jang, J.C. (2010). The Arabidopsis bZIP1 Transcription Factor Is Involved in Sugar Signaling, Protein Networking, and DNA Binding. Mol Plant 3, 361-373. <br /> <br /> Ke Y, Han G, He H, and Li J. (2009). Differential regulation of proteins and phosphoproteins in rice under drought stress. Biochem. Biophys. Res. Commun. 379: 133-138. <br /> <br /> Kiirats, O, DM Kramer, GE Edwards. 2010 Co-regulation of dark and light reactions in three biochemical subtypes of C4 species. Photosynthesis Res 105:83-88. <br /> <br /> Koch, K.E. (2010) A novel sucrose synthase pathway for sucrose degradation in cultured Sycamore cells COMMENTARY. On: Huber, S.C., and T. Akazawa. Plant Physiol. 81:1008-1013. Classics collection. (plantphysiol.org/misc/classics.dtl). <br /> <br /> Lee, S.M., Y.H. Lee, H.-u. Kim, S.-c. Seo, S.-j. Kwon, H.-s. Cho, S.-I. Kim, T. Okita, D. Kim (2010) Characterization of the potato upreg1gene, encoding a mutated ADP-glucose pyrophosphorylase large subunit, in transformed rice. Plant Cell Tissue and Organ Culture 102:171-179. DOI: 10.1007/s11240-010-9718-4.<br /> <br /> Leisner, CP, AB Cousins, S Offermann, TW Okita, GE Edwards. The effects of salinity of photosynthesis and growth of the single-cell C4 species Bienertia sinuspersici (Chenopodiaceae). Photosynthesis Res. In press. <br /> <br /> Lin, P.C., Pomeranz, M.C., Jikumaru, Y., Kang, S.G., Hah, C., Fujioka, S., Kamiya, Y., and Jang, J.C. (2010). The Arabidopsis tandem zinc finger protein AtTZF1 can affect ABA- and GA-mediated growth, stress, and gene expression responses. Plant Journal (in press). <br /> <br /> Luhua S, Ciftci-Yilmaz S, Harper J, Cushman J, Mittler R. (2008) Enhanced tolerance to oxidative stress in transgenic Arabidopsis plants expressing proteins of unknown function. Plant Physiol. 148(1): 280-92 <br /> <br /> Miller, R., Wu, G., Deshpande, R.R., Vieler, A., Gaertner, K., Li, X., Moellering, E.R., Zäuner, S., Cornish, A.J., Liu, B., Bullard, B., Sears, B.B., Kuo, M.H., Hegg, E.L., Shachar-Hill, Y., Shiu, S.H., Benning, C. (2010) Changes in transcript abundance in Chlamydomonas reinhardtii following nitrogen-deprivation predict diversion of metabolism. Plant Physiol. Online. <br /> <br /> Moellering, E. R. and Benning, C. (2010) RNA interference silencing of a major lipid droplet protein affects lipid droplet size in Chlamydomonas reinhardtii. Eukaryot. Cell 9:97-106.<br /> <br /> Morsy, M, S Gouthu, S Orchard, D Thoneycroft, JF Harper, R Mittler, JC Cushman (2008) Charting plant interactomes: possibilities and challenges. Trends in Plant Sci. 13: 183-191. <br /> <br /> Oh, M.-H., Sun, J., Oh, D.H., Zielinski, R.E., Clouse, S.D. and Huber, S.C. 201X. Enhancing photosynthesis and plant growth by engineering the BRASSINOSTEROID INSENSITIVE 1 receptor kinase. Plant Physiol., submitted. <br /> <br /> Oh, M.-H., Wang, X., Wu, X., Zhao, Y., Clouse, S.D. and Huber, S.C. 2010. Autophosphorylation of Tyr-610 in the receptor kinase BAK1 plays a role in brassinosteroid signaling and basal defense gene expression. Proc. Natl. Acad. Sci. USA 107: 17827-17832. <br /> <br /> Oliver MJ, Cushman JC, Koster KL (2010) Dehydration Tolerance in Plants. Methods in Molecular Biology: Plant Stress Tolerance. 639: 3-24. <br /> <br /> Park J, TW Okita, GE Edwards. 2010 Expression profiling and proteomic analysis of isolated photosynthetic cells of the non-Kranz C4 species Bienertia sinuspersici. Functional Plant Biology 37: 1-13. <br /> <br /> Parry, M.A.J., Reynolds, M., Salvucci, M.E., Raines, C., Andralojc, P.J., Zhu, X-G., Price, D., Condon, A. G. and Furbank, R. (2010) Raising yield potential of wheat: increasing photosynthetic capacity and efficiency. J. Exp Bot. DOI:10.1093/jxb/erq304 <br /> <br /> Penning, B.W., C.T. Hunter III, R. Tayengwa**, A.L. Eveland, C.K. Dugard, A.t. Olek, W.E. Vermeris, K.E. Koch, D.R. McCarty, M.F. Davis, S.R. Thomas, M.C. McCann, N.C. Carpita. 2009. Genetic resources for maize cell wall biology. Plant Physiol. 151:1703-1728. <br /> <br /> Pomeranz, M., Lin, P.C., Finer, J., and Jang, J.C. (2010). AtTZF gene family localizes to cytoplasmic foci. Plant Signal Behav 5, 190-192. <br /> <br /> Pomeranz, M.C., Hah, C., Lin, P.C., Kang, S.G., Finer, J.J., Blackshear, P.J., and Jang, J.C. (2010). The Arabidopsis tandem zinc finger protein AtTZF1 traffics between the nucleus and cytoplasmic foci and binds both DNA and RNA. Plant Physiol 152, 151-165. <br /> <br /> Reddy KR, Prasad PVV and Singh SK. 2010. Effects of ultraviolet-B radiation and its interaction with climate change factors on agricultural crop growth and yield. In W. Gao (eds). UV Radiation and Global Change: Measurement, Modelling and Effects on Ecosystems. Springer  Verlag, USA. pp. 395  437. <br /> <br /> Ristic Z, Momcilovic U, Bukovnik U, Prasad PVV, Fu J, DeRidder BP, Elthon TE, Mladenov N. 2009. Rubisco activase and wheat productivity under heat stress conditions. Journal of Experiment Botany 60: 4003-4014. <br /> <br /> Rumpho ME, S Pochareddy, JM Worful, EJSummer, D Bhattacharya, KN Pelletreau, MS Tyler, J Lee, JR Manhartand KM Soule 2009Molecular characterization of the Calvin cycle enzyme phosphoribulokinase in the stramenopile alga Vaucheria litorea and the plastid hosting mollusk Elysia chlorotica.Molecular Plant 2: 1384-1396.<br /> <br /> Rumpho, ME, Pelletreau, KN , Moustafa, A and Bhattacharya, D. (2010) Making of a photosynthetic animal. Journal of Experimental Biology (in press, Dec. 2010) <br /> <br /> Salvucci, M. E., Barta, C., Byers, J. A. and Canarini, A. (2010) Photosynthesis and assimilate partitioning between carbohydrates and isoprenoid products in vegetatively active and dormant guayule: physiological and environmental constraints on rubber accumulation in a semi-arid shrub. Physiol. Plantarum, DOI: 10.1111/j.1399-3054.2010.01409.x <br /> <br /> Sanjaya, Durrett, T.P., Weise S.E., Benning, C. (2010) Increasing the energy density of vegetative tissues by diverting carbon from starch to oil biosynthesis in transgenic Arabidopsis. Plant Biotech J. under review <br /> <br /> Schlauch KA, Grimplet J, Cushman JC, Cramer GR (2010) Transcriptomics analysis methods: microarray data analysis and visualization using the Affymetrix GeneChip® Vitis vinifera genome array. In: Methods in Grapevine Research. Eds: Serge Delrot. COST Book Chapter xx, pp. xxx-xxx. <br /> <br /> Silvera K, Neubig KM, Whitten WM, Williams NH, Winter K, Cushman JC. (2010a) Evolution along the Crassulacan acid metabolism continuum. Function Plant Biology. 37: 995-1010 (Cover article). <br /> <br /> Silvera K, Santiago LS, Cushman JC, Winter K. (2009) Crassulacean acid metabolism and epiphytism linked to adaptive radiations in the Orchidaceae. Plant Physiol. 149:1838-1847. <br /> <br /> Silvera K, Santiago LS, Cushman JC, Winter K. (2010b) Incidence of Crassulacean acid metabolism in the Orchidaceae derived from carbon isotope ratios: a checklist of the flora of Panama and Costa Rica. Botanical Journal of the Linnean Society. 163: 194-222. <br /> <br /> Smith DR, Lee RW, Cushman JC, Magnuson JK, Tran D, Polle JEW. (2010) The Dunaliella salina organelle genomes: large sequences, inflated with intronic and intergenic DNA. BMC Plant Biology. 10:83. <br /> <br /> Song, G.-Q, Sink, K.C, Ma, MH, T, Herlache, J.F. Hancock, Loescher 2010. A novel mannose-based selection system for plant transformation using c elery mannose-6-phosphate reductase gene Plant Cell Reports 62: 163-172.<br /> <br /> Sun, J., Zhang, J. Larue, C.T. and Huber, S.C. 201X. Decrease in leaf sucrose synthesis leads to increased leaf starch turnover and decreased RuBP-regeneration-limited photosynthesis but not Rubisco-limited photosynthesis in Arabidopsis mutants of SPS. Plant Cell Environ, in press. <br /> <br /> Sunagawa H, Cushman JC, Agarie S. (2010) Crassulacean acid metabolism alleviates reactive oxygen species in the facultative CAM plant, the common ice plant, Mesembryanthemum crystallinum. Plant Production Science. 13: 246- 260. <br /> <br /> Tillett RL, Cushman JC (2010) Vitis functional genomics: Open systems for transcriptome analysis. In: Encyclopedia of Plant Genomics. Eds: José M. Martinez-Zapater and Anne-Francoise Adam Blondon. Science Publishers, Enfield, New Hampshire. Chapter 11, pp. xxx-xxx. <br /> <br /> Ting L, Huang, S, Jiang, WZ, Wright, D, Spalding, MH, Weeks, MH, Yang, B. (2010) TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA- cleavage domain. Nucleic Acids Research 39: 359-372.<br /> <br /> Voznesenskaya, EV, NK Koteyeva, GE Edwards, G Ocampo. 2010 Revealing diversity in structural and biochemical forms of C4 photosynthesis and a C3C4 intermediate in genus Portulaca L. (Portulacaceae). J Exp Bot. 61: 3647-3662. <br /> <br /> Wang, L, Fan, l, Loescher, W, Duan, W, Cheng, J, Liu, G, Li , S. (2010) Salicylic acid alleviates decreases in photosynthesis under heat stress and accelerates recovery in grapevine leaves BMC Plant Biology 10:34. <br /> <br /> Wu, X., Oh, M.-H., Schwarz, E.M., Larue, C.T., Sivaguru, M., Imai, B.S., Yau, P.M., Ort, D.R. and Huber, S.C. 201X. Non-histone lysine acetylation is an abundant protein modification in Arabidopsis. Plant Physiol., in revision. <br /> <br /> Wu, X., Wang J., Na J.-K., Yu Q., Moore R.C., Zee F., Huber, S.C. and Ming, R. 2010. The origin of the non- recombining region of sex chromosomes in Carica and Vasconcellea. Plant J., 63: 801-810. <br /> <br /> Zell, MB, H Fahnenstich, A Maier, M Saigo, EV Voznesenskaya, GE Edwards, C Andreo, F Schliefenbaum, C Zell, MF Drincovich and VG Maurino. 2010 Analysis of Arabidopsis with highly reduced levels of malate and fumarate sheds light on the role of these organic acids as storage carbon molecules. Plant Physiol. 152: 1251-1262. <br /> <br /> Zinn, KE, M Tunc-Ozdemir, JF Harper (2010) Temperature stress and plant sexual reproduction: uncovering the weakest links. J Exp Bot 61: 1959-1968.<br /> <br />

Impact Statements

1. Characterization of this long-term animal-plastid association is providing information on the requirements for nuclear-cytosolic interactions to sustain plastid structure and function, as well as the synthesis and transfer of carbon compounds from the captured organelle to the host. Ultimately, this may add to the information necessary for maintaining these energy-capturing organelles in culture or foreign hosts for long periods of time
2. These regions may serve as targets for either the design of an improved Rubisco, or for genetic selection following site-directed or random mutagenesis. Since Rubisco presents an obvious target for increasing photosynthetic performance these studies lay the groundwork for understanding this major player in photosynthesis.
3. Studies of Obj. C. on rutabaga and algae have shown that the conversion of photosynthetic sugars into triacylglycerols will be useful for the engineering of novel biofuel crops and as feed stocks for biofuel production. The data presented for the artificial TALN modification system shows promise in producing targeted gene disruption in plants and algae and, potentially, for gene replacement by homologous recombination.
4. Understanding the factors inhibiting photosynthesis when heat stress is imposed under natural conditions is essential for developing strategies to maintain plant yields under climate change. In addition, studies on basic factors, such as the described histone methyltransferase and calcium signaling proteins improve our understanding of regulation of plant development and pathways plants use to regulate growth. These studies bear directly on future strategies to improve photosynthate partitioning and grain filling.

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Report Information

Participants

Christoph Benning, MSU AgBioResearch;
Julie Stone, Nebraska AES;
Gerry Edwards, Washington AES;
Marty Spalding, Iowa AES;
Don Weeks, Nebraska AES;
Steve Huber, USDA-ARS, Illinois;
JC Jang, Ohio AES;
Glenda Gillaspy, Virginia AES;
John Cushman, Nevada AES;
Wayne Loescher, MSU AgBioRsearch;
Tom Sharkey, MSU AgBioResearch;
Laura Gentry, Illinois AES;
Robert Aiken, Kansas AES;
Mike Salvucci, USDA-ARS, Arizona;
Irwin Goldman, AA, Madison, Wisconsin
Steve Pueppke, Director MSU AgBioResearch

Brief Summary of Minutes

Meeting Minutes

Outcome of the Business meeting:

1. Jeff Harper and John Cushman (Nevada AES) will host the annual meeting in 2012 in Reno Nevada

2. Marty Spalding and Steve Rodermel (Iowa AES) will organize the annual meeting in 2013 possibly together with Julie Stone (Nebraska AES) in Omaha, NE.

3. The renewal proposal has been completed and will be submitted on time by Christoph Benning on Dec 2011. We expect to hear following the NC AES directors meeting in March or April 2012

4. Christoph Benning will take over from Irwin Goldman as Administrative Advisor and will be supported by MSU AgBioResearch Director Steve Pueppke.

5. It was mentioned by the AA that for the SAES422 Reprort form, funding information is needed.

6. Laura Gentry (Illinois AES), was elected as new member.

Agenda of the Meeting:

Friday, Nov. 11, 2011

Evening
7:30 Meet in Gatehouse Lobby (Transport by minivan)
8:00 Dinner at Dustys Cellar
10:00 Return to Gatehouse (Transport by minivan)

Saturday, Nov. 12, 2011

Morning
7:45 Meet in Gatehouse Lobby (Transport by minivan)
8:00 Breakfast in MSU Biomedical and Physical Science Building (BPS)

8:45 Announcements and introductions, Christoph Benning
9:00 Welcome by MSU AgBioResearch Director Steve Pueppke.
9:15 Christoph Benning, MSU AgBioResearch
9:45 Julie Stone, Nebraska AES
10:15 Gerry Edwards, Washington AES
10:45 Break
11:00 Marty Spalding, Iowa AES
11:30 Don Weeks, Nebraska AES

Afternoon
12:00 Lunch and Business Meeting I, 1400 BPS

1:00 Steve Huber, USDA-ARS, Illinois
1:30 JC Jang, Ohio AES
2:00 Glenda Gillaspy, Virginia AES
2:30 Break
2:45 John Cushman, Nevada AES
3:15 Wayne Loescher, MSU AgBioRsearch
3:45 Tom Sharkey, MSU AgBioResearch
4:15 Break
4:30 Laura Gentry, Illinois AES
5:00 Robert Aiken, Kansas AES
5:30 Mike Salvucci, USDA-ARS, Arizona
6:00 Business Meeting II

Evening
6:30 Return to Gatehouse (Transport by minivan)
7:30 Meet in Lobby of Gatehouse (Transport by minivan)
8:00 Dinner at Bennings
11:00 Return to Gatehouse (Transport by minivan)

Accomplishments

Summary of Accomplishments Presented by Meeting Attendees<br /> <br /> Objective 1. Plastid Function and Intracellular Communication <br /> <br /> Christoph Benning (MSU AgBioresearch) presented new findings on chloroplast lipid remodeling and plant stress tolerance related to Objective 1 of the renewal proposal. Lipid composition of cellular membranes is affected by phosphate deprivation leading to the replacement of phospholipids with non-phosphorus glycolipids synthesized at the chloroplast outer envelope membrane. A novel galactolipid remodeling enzyme was described, which is involved in the protection of plants against freezing stress. Activation of this enzyme leads to the formation of oligogalactolipids and triacylglycerols allowing membranes to adjust as the chloroplast shrinks following severe dehydration during freezing and ice formation.<br /> <br /> Julie Stone (Nebraska AES), a new member, talked about her work with Fumonisin B1 (FB1) a sphingolipid analogue that inhibits ceramide synthases in both plants and animals, and thereby induces programmed cell death. Julie and her coworkers isolated a series of FB1 resistant mutants that she is now studying. One affects a transcription factor causing a change in sphingolipid profiles. Julie also presented her work on Arabidopsis DJ-1 like proteins, which are conserved ubiquitous proteins. A human ortholog is implicated in Parkinsons disease (Park7) and mutations destabilize the dimer. The protein is also overproduced in various tumors. Arabidopsis has three different forms (A, B, and C). AtDJ1C, the current focus of her research, is localized in the plastid. The gene is expressed mostly in young leaves. Homozygous mutants are not viable, but can be rescued with sucrose supplementation and an epitope-tagged version of the wild-type DJ1C protein. DJ1C-interacting partners will be identified using coimmunoprecipitation of DJ1C-containing complexes from chloroplasts isolated from the complemented line. She is proposing to interact with the Benning lab on chloroplast isolation techniques. She will apply omics technologies to characterize these proteins. Two University of Nebraska - Lincoln NSF REU programs were described that contribute to training of undergraduate students involved in her research.<br /> <br /> Objective 2. Photosynthetic Capture and Photorespiratory Release of CO2<br /> <br /> Gerry Edwards presented collaborative work with Asaph Cousins and R. Giuliani (Washington AES) on efforts at overcoming the limitations of C3 photosynthesis, photoinhibition, and sink limitation. The group studies photorespiration in rice and proposes to redesign photosynthesis in this crop. In the long term, it is planned to introduce C4 photosynthetic metabolism into rice. Natural variations in leaf structure and gas exchange in different wild rice species were analyzed and correlated to the diversity in water use and sun versus shade adaptations. One of the wild rice species, Oryzacoarctata, was found to have a cellular arrangement of mesophyll and bundle sheath cells like that in some Kranz type C4 grasses. In some species the mesophyll cells have lobes in which mitochondria are located in close proximity to the chloroplasts. The group is investigating the significance of these lobes to determine whether they function to facilitate diffusion of atmospheric CO2 to chloroplasts, or to enhance refixation of photorespiredCO2 like C3-C4 intermediates. The CO2 compensation point in plants is a function of Rubisco specificity factor and degree of refixation of photorespiredCO2. In vitro analysis of the specificity factor will be combined with gas exchange to enable modeling of the response of photorespiration to different temperatures. Approaches to measure the CO2 compensation point in vivo and techniques to measure the internal rate of refixation of CO2 using stable isotopes were presented. <br /> <br /> Marty Spalding, (Iowa AES) presented his newest findings on the carbon concentration mechanism (CCM) in Chlamydomonas. CCM is inducible at low CO2 and there are multiple acclimation states at low and very low CO2 concentrations. Interestingly, one mutant, lcib, does not survive low CO2 concentrations, but is viable under very low CO2 concentrations. His group conducted transcriptomics work using the cia5 mutant and different CO2 concentrations. CIA5(CCM1) is a transcription factor acting as master regulator of CCM. He observed 16 clusters of expressed genes with similar behavior and described examples. For example, cluster 15 contained genes that are induced under low CO2. He also described attempts at ectopic expression of LCIA and LCIB under the control of the Rcbs promoter to increase biomass yields in reactors. Overexpression of both genes caused an increase in final biomass yield. Growth remained the same but the mass of the cells was increased through increase in starch content (60% per DW). The cells showed an increased CO2 assimilation. Crossing the transgenics with the starch deficient mutant sta6 led to increased total fatty acids but decreased biomass yield.<br /> <br /> Don Weeks (Nebraska AES) also reported on the global transcriptional regulation of CCM by CO2 deprivation, in this case in the wild type. 25% of genes were found to be differentially expressed 4 fold or greater. It was noted that the nature of the CO2 shift is important, e.g., when using a fermenter switching from 4% CO2 to ambient CO2 levels it takes an hour to complete the shift. He also described different clusters of co-regulated genes. For example, photosynthetic genes were strongly reduced under these conditions. It was observed that most genes organized head-to-head (HTH) in the genome are co-regulated by bidirectional promoters. Don provided two examples of HTH gene pairs encoding CCM components that are coordinately expressed at high levels. A search for common TF binding sites identified a CACACA&.motif in up regulated genes, but not in down regulated genes.<br /> <br /> John Cushman (Nevada AES) described his omics studies on ice plant, in which crassulacean metabolism (CAM) is inducible through salinity or drought stress. CAM plants have about 20% the crop water demand of C3 plants, but can have higher rates of above ground productivity. His lab compared the induced and non-induced states by global transcription profiling. He described an ice plant mutant deficient in CAM that grows more slowly than wild type, lacks nocturnal CO2 uptake and starch. The CAM mutant contains a knockout in the plastidic phosphoglucomutase gene. John described a metabolic profiling study during day-night period following constant conditions of illumination and temperature to evaluate the role of circadian clock regulation of CAM outputs. Rhythmic acid formation following drought stress was observed in the wild type, but not the mutant. Starch was transiently present in wild type, but absent from the mutant. Starch content remained high under continuous conditions. Soluble sugars were found to cycle in the mutant, which was interpreted as a compensatory effect. The metabolite analysis was done commercially (Metabolon). 503 metabolites were identified including 199 known and 304 unnamed compounds. The accumulation of free fatty acids and monoglycerides was observed in the mutant. The mutant showed a lower ABA content and evidence of higher oxidative stress. A reciprocal pattern for reduced and oxidized glutathione was observed over time.<br /> <br /> Objective 3: Mechanisms Regulating Photosynthate Partitioning<br /> <br /> Steve Huber (Ilinois ARS) reported on his work on brassinosteroid signaling in particularly the mechanism of deactivation. He focused on the BRI1 receptor kinase, and its auto and transphosphorylation properties. The BRI1 activation loop of the kinase domain becomes phosphorylated during activation. BRI1 is deactivated by dephosphorylation and endocytosis, or phosphorylation of inhibitory sites and calmodulin interaction. He reported on work focusing on the phosphorylation of Ser 891. This phosphorylation occurs in vivo and attenuates brassinosteroid signaling. A phosphomimetic mutant is unresponsive. He reported that transgenic plants with substitutions at Ser 891 have growth phenotypes, such as enhanced growth resulting in increased biomass. He noted that BRI1 binds to immobilized calmodulin (CaM) with its kinase domain. He discovered that CaM affects autophosphorylation of BRI1 and transphosphorylation of E. coli proteins if coproduced in E. coli. In essence, this observation provides the basis for a new assay of BRI1 activity.<br /> <br /> JC Jang (Ohio, AES) covered work from his lab on sugar signaling. His focus is on sugar responsive Transcription Factors (TFs) such as bZIP1, which is a negative regulator for plant growth in the absence of sugar. He reported on the dimerization of different TFs of the bZIP family. He also reported on TZF1 (Tandem Zinc Finger1). Interestingly, only 10-20% of TZF1-GFP fusion protein was found in nucleus, while most of it was found in punctate patterns in cytosol. These punctate structures were identified as processing (P) bodies and stress granules, which are not the same but have overlapping composition. Both can be induced by stress. The Arabiopsis AtTZF gene family has 11 members. All localize to the nucleus as well as to P-bodies and stress granules as demonstrated by co-localization of GFP fusion proteins and specific marker proteins for P-body and stress granule. He reported that TZF1 is a positive regulator of sugar/ABA signaling and a negative regulator for GA signaling. The expression of the C-terminal fragment of TZF1 causes dominant negative effects and enhances growth, but the plants are sensitive to drought. These findings might provide a strategy to enhance growth of crops such as soybean.<br /> <br /> Glenda Gillaspy (Virginia AES) reported on the regulation of the energy sensor SnRK1. It is involved in a mechanism that helps plants to respond to low carbon (low energy) conditions. Arabidopsis has three SnRK1 genes. Overexpression leads to gain of function (more biomass) through metabolic reprogramming. On the contrary, RNAi plants were small. Spatial regulation was studied by promoter GUS fusion showing that 1.2 has more restricted expression than 1.1. Her group discovered that the P80 DWD domain protein of previously unknown function interacts directly with SnRK1. Mutants lacking P80 show similar phenotypes to SnRK1 RNAi plants. However, SnRK1.1 overproduction could rescue P80 mutants. The level of SnRK1.1 was found to be low in P80 mutants. SnRK1.1 normally localizes to the cytosol but in the P80 mutant background it was associated with the membrane. Glenda speculated that P80 might form different types of complexes as the expression of 1.2 in the P80 mutant background led to mislocalization.<br /> <br /> Tom Sharkey (MSU AgBioResearch), a new member, discussed his work defining limitations of photosynthesis with regard to different fates of photosynthate during the day (formation of sucrose) and night (break down of transitory starch formed during the day). He introduced the concept of triosephosphate limitation of photosynthesis. He compared the Arabidopsis tpt mutant, which cannot export carbohydrates during the day, and the starch deficient plastid pgm mutant affected in export of sugars from the plastid during the night. Emphasis was on the role of the proton motive force (PMF). CO2 response curves were determined to be similar at different temperatures. PMF was found to decline at higher CO2 but not at lower temperatures. In the tpt mutant PMF increased with increasing CO2. Tom noted that sucrose and starch syntheses are sensitive to phosphate concentration (inhibitory) and that triosephosphate use (TPU) limitation is balanced by inhibitory effects of Pi and the need for ATP synthesis. Tom pointed out that sucrose production during the day does not need hexokinase while carbohydrate metabolism during the night does. Accordingly, sugar sensing and hexokinase mediated signaling should differ during the night and the day. He reported that down regulation of glucan water dikinase, an enzyme which adds phosphoryl groups to starch and promotes starch break down, increases starch content. Analyzing such a transgenic line, particularly when using inducible promoters, should provide a new approach to study the effect of carbohydrate utilization on photosynthetic capacity in plants.<br /> <br /> Objective 4: Developmental and Environmental Limitations to Photosynthesis<br /> <br /> Wayne Loescher (MSU AgBioResearch) showed the results of a large scale global analysis of three transgenic Arabidopsis lines overexpressing three alternative abiotic stress resistance genes: CBF/DREB1 (low temp regulation), M6PR (mannitol synthesis), SOS1 (sodium proton antiporter). These lines were exposed to increasing salt stress. Growth was evaluated and global transcript analyses were conducted. The transgenic lines showed fewer gene expression changes than the wild type - perhaps indicative of salt tolerance. As mannitol serves as a fungal evasion signal, those respective transgenic plants seemed to upregulate biotic stress pathways. In addition, multigenerational studies in the field are under way to test competitiveness and fitness effects.<br /> <br /> Laura Gentry (Illinois AES), a new member reported on her work done in collaboration with Fred Below. She conducts field scale research investigating yield penalty of corn during continuous growth and N-fertilization compared to growth under crop rotation conditions. The field trial is currently in its seventh year and the results demonstrated that the yield penalty under continuous conditions increases over the years, but can be compensated by increased N-fertilization. Laura discussed the effects of corn residue and the loss of N into bacterial biomass. She noted that autotoxicity could be involved as corn releases compounds which inhibit its own growth in the next generation. She discussed best practices to achieve agricultural sustainability in view of increasing demands for the production of food and fuels.<br /> <br /> Rob Aiken (Kansas, AES) presented results of field studies conducted by Vara Prassad on high temperature stress, and the basis of tolerance or susceptibility to drought in soy bean. Pollen viability in response to temperature stress as well as ROS formation and membrane damage were discussed. Rob also described his own field studies on Sorghum under semiarid conditions in Western Kansas. He described methods to determine transpiration and stomatal conductance under field conditions using image analysis. Biomass, and light-use and water-use efficiency were considered. It was concluded that if light use efficiency is increased water-use efficiency increased as well.<br /> <br /> Mike Salvucci (Arizona-ARS) described his studies on improving RuBisco Activase to achieve higher thermotolerance of photosynthesis. He reported on biochemical approaches to study Rubisco activase structure function relationships. Rubisco activase belongs to the group of AAA+ proteins, which typically remodel other proteins. He reported on the crystallization of the alpha helical domain. This enabled modeling the interaction between RuBisCo activase and RuBisCo and allowed binding competition studies with the alpha helical domain fragment. He is working on the introduction of a heat stable activase into Camelina in a collaborative project with John Cushman. Mike also reported on remote sensor studies of cotton, during which the relationship between heat stress and drought stress is apparent under field conditions. This process is hard to simulate under growth chamber conditions demonstrating the unique value of these field studies.<br /> <br /> CURRENT FUNDING <br /> <br /> Steve Rodermel<br /> <br /> DOE, DE-FG02-10ER20147. The immutans variegation mutant of Arabidopsis. $510,000 (10/01/10-7/31/13).<br /> <br /> NSF (UCSD PO 10301097-001). Regulation of chloroplast gene expression. $195,000. (07/01/09-06/30/12)<br /> <br /> UISFL. Global pathways for educating students in biodiversity. $179,000 (10/01/09- 9/30/12)<br /> <br /> Christoph Benning<br /> <br /> NSF MCB 0741395. Mechanisms of lipid trafficking between the endoplasmic reticulum and the chloroplast. $546,818 (05/01/08-04/30/11)<br /> <br /> DOE, DE-FG02-98ER20305. Regulation of Thylakoid Lipid Biosynthesis in Plants. $560,000 (08/01/10-7/31/13<br /> <br /> DOE,GLBRC. Multi-investigator project, T. Donahue PI. Funds to CB~$200,000 per year based on annual renewal (09/01/07-08/31/12). Regulation of oil biosynthesis in vegetative tissues and the engineering of biofuel crop plants producing oil in roots and shoot tissues.<br /> <br /> Aurora Biofuels. Identification of lipid switches in microalgae $250,000 (10/01/2008-09/30/2011).<br /> <br /> US AFOSR. Regulation of oil biosynthesis in algae. ($561,382 (09/01/11-08/31/14).<br /> <br /> Cushman, John C.<br /> <br /> NSF IOS-084373. Regulatory and signaling mechanisms of crassulacean acid metabolism: A photosynthetic adaptation of environmental stress. $988,389 (8/15/09-8/14/12) <br /> <br /> NSF DBI 0741876. Research Coordination Network (RCN) Program. RCN: An international research and education collaboration for grape functional genomics. $509,420 (9/15/08-9/14/13) <br /> <br /> Hatch NEV-00372. Improved abiotic stress tolerance of Camelina: A Novel Biofuel Crop for Nevada. $529,900 (7/1/10-6/30/15)<br /> <br /> DOE, DE-EE0000272. Nevada Renewable Energy Consortium: Municipal Wastewater for Microalgae Biofuel Feedstock Production. $250,000 (10/1/10-9/30/11)<br /> <br /> DOE, SBIR Phase II DE-SC0001306. High efficiency microalgae biofuel harvest and extraction using ionic liquids. $60,766 (10/1/10-9/30/12)<br /> <br /> Gerry Edwards<br /> <br /> IRRI (Bill and Melinda Gates Foundation). Creating the Second Green Revolution by supercharging photosynthesis: C4 Rice (PI, part of a consortium under the leadership of IRRI, The Philippines; co-PI A. Cousins). $334,959 (10/01/08-04/31/12)<br /> <br /> CRDF. Identifying mechanisms for increasing carbon acquisition and water use efficiency of plants with climate change. (PI, co-PI A Cousins and Russian investigators) $54,886 (9/1/10-9/1/12) <br /> <br /> USDA. Improving Plant Productivity by Altering Nitrogen Transport Processes. (Co-PI, PI M. Tegeder) $350,000 (12/15/09-12/14/12)<br /> <br /> USDA. Biosecurity Special Research Grant Aegilops cylindrical. (Co-PI, M. Kahn PI). $28,500 funding available to Edwards (Exp. 7/31/12)<br /> <br /> Glenda Gillaspy<br /> <br /> NSF Collaborative Research: Diphospho- and Triphospho-Inositol Phosphates in Plants (PI: Gillaspy) Total Award Amount: $673,426 Total Award Period Covered: 1/1/11- 12/31/14<br /> <br /> NSF RET Supplement: Plant Cell Communication and Inositol Trisphosphates Funding Agency: NSF Budget Total: 9,400.00 Start Date: 06/01/2011 End Date: 07/29/2011<br /> <br /> Steve Huber<br /> <br /> United Soybean Board, BBI Oil Project #7222. $40,000 annual (3/1/10-2/29/13)<br /> <br /> NSF, MCB-1021363. Arabidopsis 2010: Protein interacting networks and site-specific phosphorylation in leucine-rich repeat receptor-like kinase function. Multi-investigator project, S.C. Clouse, PI. Funds to SH lab ~$200,000 per year based on annual renewal (9/1/2010-8/30/2014).<br /> <br /> NSF, IOS-1022177. Regulation of receptor kinase signaling by tyrosine phosphorylation and calmodulin binding. $499,166 (9/01/2010-8/30/2013).<br /> <br /> Jeff Harper<br /> <br /> NSF, MCB-0920624. Calcium Dependent Protein Kinases in Pollen Tube Tip Growth. $390.000 (09/01/09-08/31/12)<br /> <br /> DOE, DE-FG03-94ER20152. P-type ATPases in Plants  Role of Lipid Flippases in Membrane Biogenesis $390,000 (09/01/09-08/31/12).<br /> <br /> NIH, 1RO1 GM070813-01. Ca2+-Pumps in Plants: Pollen Growth and Fertilization. $954,000 (04/01/04-08/31/12).<br /> <br /> JC Jang<br /> <br /> NSF IOB-053751. The role of AtbZIP1 in sugar regulatory network. $480,000 (02/01/06-01/31/10).<br /> <br /> OHOA1387. Plant peptide hormone technology in enhancing biomass and fitness to environmental stresses. $50,000 (03/01/2009-04/30/2012).<br /> <br /> OHOA0794. The novel roles of tandem zinc finger proteins in multi-stress tolerance in plants. $60,000 (07/01/2010-06/30/2012).<br /> <br /> Karen Koch<br /> <br /> USDA-NRI- Plant Biochemistry (07-03580). Roles and regulation of sorbitol metabolism in maize. $394,307 (09/01/07  08/31/12).<br /> <br /> NSF- Plant Genome (08-21952). Functional genomics of transfer cells. Funds to Koch lab ~$150,000 per year (09/09  8/12).<br /> <br /> BSF- Plant Genome (IOS-1025976). GEPR/LIT Genetic and genomic approaches to understanding long-distance transport and carbon partitioning. Funds to Koch lab ~$140,000 per year (12/10  11/14).<br /> <br /> BSF- Plant Genome (IOS-1116561). UniformMu: A transposon resource for mutagenesis in maize. Funds to Koch lab ~$200,000 per year based on annual renewal (09/11  08/13). <br /> <br /> USDA-NIFA (2011-67003-30215). Adapting kernel metabolism to enhance cereal yield under adverse conditions. Funds to Koch lab ~$100,000 per year based on annual renewal (08/11  07/14)<br /> <br /> David Kraemer<br /> <br /> NIH, (2 RO1 GM061904) Rieske Headgroup Cytochrome Complexes - Metal Ion Probes Membrane Protein Structure and Function (Co-PI with Dr. Michael Bowman) $2,057,830 (1/1/09-12/31/12)<br /> <br /> DOE (DE-FG02-04ER15559) The Energy Budget of Steady-State Photosynthesis $540,000 (7/1/08-6/30/12) <br /> <br /> USDA, Plant Biology (C): Biochemistry (2008-35318-04665) Co-regulation of the light and dark reactions of photosynthesis. $400,000 (1/01/09-12/31/12)<br /> DOE (DE-EE0003046) The National Alliance for Advanced Biofuels and Bioproducts an Algal Biofuels Consortium (Lead Consortium Lead: The Donald Danforth Plant Science Center, Executive Director: Jose Olivares, Los Alamos National Lab and Donald Danforth Plant Science Center). $54,000,000 ($660,000 to MSU) (1/01/10-12/31/13)<br /> <br /> DOE Interdisciplinary Research and Training in the Plant Sciences ($240,000 direct funds) (06/16/2010-08/31/2011)<br /> <br /> NSF. (RC100150) Plug and Play Photosynthesis for RuBisCO Independent Fuels $3.6M for 7 co-P.I.s, $300,599 to MSU (06/01/2011-05/31/2014)<br /> <br /> DOE. Center for Advanced Camelina Oil (CECO) 9.3M ($860,000 to MSU, in negotiation), 1/1/12-12/31/15<br /> <br /> Jiaxu Li<br /> <br /> USDA-(NIFA) 2009-04041 Role of histone H3 lysine 36 methylation in regulating developmentally important genes in rice. $134,815 (01/01/10 ? 12/31/11)<br /> <br /> Wayne Loescher<br /> <br /> USDA FAS 58-3148-8-159 Enhancing salinity tolerance in canola $60,000 (9/1/09-8/31/12)<br /> <br /> USDA-CSREES/NIFA 2010-85117-20570 Building Expertise in Plant Breeding that Focuses on Drought Tolerance $500,000 (2/1/10  1/31/14)<br /> <br /> Mike Salvucci<br /> <br /> DOE, DE-AI02-97ER20268. Optimizing Rubisco Regulation for Increased Photosynthetic Performance under Climate Change. $381,000 (05/01/2010-04/30/2012).<br /> <br /> Robert Spreitzer<br /> <br /> DOE, DE-FG02-00ER15044. Role of the Rubisco Small Subunit. $495,000 (05/01/10-04/30/13).<br /> <br /> NSF, EPSCoR. Nebraska Center for Algal Biology and Biotechnology. Multi-investigator project. Funds to my lab ~$306,322 (10/01/10-09/30/15).<br /> <br /> Thomas D. Sharkey<br /> <br /> NSF, IOS-0950574.Function and regulation of isoprene synthesis in leaves. $590,000 (02/15/11-01/31/14)<br /> <br /> ZuvaChem Inc. Novel isoprene synthase genes for isoprene production.$300,000. (07/01/11-06/30/14)<br /> <br /> Julie Stone<br /> <br /> NSF, DBI-1156692. NSF REU SITE: Training in Redox Biology. PI: Don Becker; co-PI: Julie Stone. $278,500 (4/12-3/15)<br /> <br /> Univ. Nebraska IANR. Novel Subunits of Serine Palmitoyltransferase, New Players in Sphingolipid Synthesis PI: Ed Cahoon, co-PI: Julie Stone., $60,000 (05/01/2010-04/40/2012)<br /> <br /> ASPB-SURF. Arabidopsis thaliana Mutants Defective in DJ-1 Homologs. Julie Stone/Amanda Leafgren. $5275 (05/01/2010-04/30/2011)<br /> <br /> Univ. Nebraska IANR. Plant Stress Biology Initiative Co-PIs: Julie Stone, Gary Brewer, $15,000 (05/2010-04/2012.<br /> <br /> NSF-DBI-0851747. NSF REU SITE: Training in Redox Biology. PI: Don Becker; co-PI: Julie Stone. $252,250 (4/09-3/12)<br /> <br /> <br /> Don Weeks 2011<br /> <br /> NSF, MCB-0952533 Bicarbonate Transport in Chlamydomonas $540,000 (3/15/2010  02/28/2013)<br /> <br /> NS, EPSCoR-1004094 Building Infrastructure in Nanohybrid Materials and Algal Biology Research $412,000 (10/01/2010  09/30/2015)<br /> <br /> DOE, DE-EE0001052 Renewable Biofuels from Algae $200,000 (02/01/2010  01/31/2012)<br /> <br /> DOE  CAB-COMM DE-EE0003373 Consortium for Commercialization of Algae Biofuels and Biotechnology $300,000 (09/01/2010  08/31/2013)<br />

Publications

Publications 2010/2011<br /> <br /> 1. Ananda, N., P. V. Vadlani, and P. V. V. Prasad. 2011. Evaluation of drought and heat stressed grain sorghum (Sorghum bicolor) for ethanol production. Industrial Crops and Products 33:779-782<br /> <br /> 2. Buescher, E., T. Achberger, I. Amusan, A. Giannini and others. 2010. Natural genetic variation in selected populations of Arabidopsis thaliana is associated with ionomic differences. PLoS One 5:e11081<br /> <br /> 3. Carmo-Silva, A. E. and M. E. Salvucci. 2011. The activity of Rubisco's molecular chaperone, Rubisco activase, in leaf extracts. Photosynth.Res. 108:143-155<br /> 4. Carmo-Silva, A. E., L. Marri, F. Sparla, and M. E. Salvucci. 2011. Isolation and compositional analysis of a CP12-associated complex of calvin cycle enzymes from Nicotiana tabacum. Protein Pept.Lett. 18:618-624<br /> <br /> 5. Castruita, M., D. Casero, S. J. Karpowicz, J. Kropat and others. 2011. Systems biology approach in Chlamydomonas reveals connections between copper nutrition and multiple metabolic steps. Plant Cell 23:1273-1292<br /> <br /> 6. Chan, Z., P. J. Bigelow, W. Loescher, and R. Grumet. 2011. Comparison of salt stress resistance genes in transgenic Arabidopsis thaliana indicates that extent of transcriptomic change may not predict secondary phenotypic or fitness effects. Plant Biotechnol.J.<br /> <br /> 7. Chan, Z., R. Grumet, and W. Loescher. 2011. Global gene expression analysis of transgenic, mannitol-producing, and salt-tolerant Arabidopsis thaliana indicates widespread changes in abiotic and biotic stress-related genes. J.Exp.Bot. 62:4787-4803<br /> <br /> 8. Cushman, J. C. and M. J. Oliver. 2011. Understanding vegetative desiccation tolerance using integrated functional genomics approaches within a comparative evolutionary framework, p. 307-338. In: U. Luttge, E. Beck, and D. Bartels (eds.), Ecological Studies: Desiccation Tolerance in Plants. Springer, New York, NY.<br /> <br /> 9. Deluc, L. G., A. Decendit, Y. Papastomoulis, J.-M. Merillon and others. 2011. Water deficit increases stilbene metabolism in Cabernet Sauvignon berries. J.Agric.Food Chem. 59:289-297<br /> <br /> 10. Dillard, S. R., K. Van, and M. H. Spalding. 2011. Acclimation to low or limiting CO(2) in non-synchronous Chlamydomonas causes a transient synchronization of the cell division cycle. Photosynth.Res 109:161-168<br /> <br /> 11. Djanaguiraman, M., P. V. V. Prasad, D. L. Boyle, and W. T. Schapaugh. 2011. High-temperature stress and soybean leaves: Leaf anatomy and photosynthesis. Crop Science 51:2125-2131<br /> <br /> 12. Djanaguiraman, M., P. V. V. Prasad, and K. Al-Khatib. 2011. Ethylene perception inhibitor 1-MCP decreases oxidative damage of leaves through enhanced antioxidant defense mechanisms in soybean plants grown under high temperature stress. Environmental and Experimental Botany 71:215-223<br /> <br /> 13. Duanmu, D. and M. H. Spalding. 2011. Insertional suppressors of Chlamydomonas reinhardtii that restore growth of air-dier lcib mutants in low CO(2). Photosynth.Res. 109:123-132<br /> <br /> 14. Edwards, G. E. and E. V. Voznesenskaya. 2011. C(4) photosynthesis: Kranz forms and single-cell C(4) in terrestrial plants, p. 29-61. In: A. S. Raghavendra and R. F. Sage (eds.), C(4) Photosynthesis and Related CO(2) Concentrating Mechanisms Advances in Photosynthesis and Respiration., vol. 32. Springer, Dordrecht.<br /> <br /> 15. Foudree, A., M. Aluru, and S. Rodermel. 2010. PDS activity acts as a rheostat of retrograde signaling during early chloroplast biogenesis. Plant Signal.Behav. 5:1629-1632<br /> <br /> 16. Genkov, T., M. Meyer, H. Griffiths, and R. J. Spreitzer. 2010. Functional hybrid rubisco enzymes with plant small subunits and algal large subunits: engineered rbcS cDNA for expression in chlamydomonas. J Biol Chem 285:19833-19841<br /> <br /> 17. Gibson, K., J. S. Park, Y. Nagai, S. K. Hwang and others. 2011. Exploiting leaf starch synthesis as a transient sink to elevate photosynthesis, plant productivity and yields. Plant Science 181:275-281<br /> <br /> 18. Gore, M. A., G. Coyle, B. Friebe, T. A. Coffelt and others. 2011. Complex Ploidy Level Variation in Guayule Breeding Programs. Crop Science 51:210-216<br /> <br /> 19. Goren, S., S. C. Huber, and D. Granot. 2011. Comparison of a novel tomato sucrose synthase, SlSUS4, with previously described SlSUS isoforms reveals distinct sequence features and differential expression patterns in association with stem maturation. Planta 233:1011-1023<br /> <br /> 20. Gross, J., K. N. Pelletreau, A. Reyes-Prieto, M. E. Rumpho, and D. Bhattacharya. 2011. Secondary and tertiary endosymbiosis: Plastid evolution and loss, plastid protein import, and photosynthesis in the sea slug Elysia chlorotica, In: R. Bock and V. Knoop (eds.), Advances in Photosynthesis and Respiration - Genomics of Chloroplasts and Mitochondria., vol. In press. Springer.<br /> <br /> 21. Henderson, J. N., A. M. Kuriata, R. Fromme, M. E. Salvucci and others. 2011. Atomic resolution X-ray structure of the substrate recognition domain of higher plant Rubisco activase. J Biol Chem<br /> <br /> 22. Huber, S. C., W. M. Kaiser, and V. Jain. 2011. Post Translational Regulation of Nitrate Reductase, p. 21-44. In: V. Jain and A. Kumar (eds.), Nitrogen Use Efficiency in Plants. New India Publishing, New Delhi.<br /> <br /> 23. Kang, S. G., J. Price, P. C. Lin, J. C. Hong and others. 2010. The Arabidopsis bZIP1 transcription factor is involved in sugar signaling, protein networking, and DNA binding. Mol.Plant 3:361-373<br /> <br /> 24. Kaye, Y., Y. Golani, Y. Singer, Y. Leshem and others. 2011. Inositol polyphosphate 5-phosphatase7 regulates the production of reactive oxygen species and salt tolerance in Arabidopsis. Plant Physiol 157:229-241<br /> <br /> 25. Koteyeva, N. K., E. V. Voznesenskaya, J. O. Berry, S. D. Chuong and others. 2011. Development of structural and biochemical characteristics of C(4) photosynthesis in two types of Kranz anatomy in genus Suaeda (family Chenopodiaceae). J Exp.Bot. 62:3197-3212<br /> <br /> 26. Koteyeva, N. K., E. V. Voznesenskaya, E. H. Roalson, and G. E. Edwards. 2011. Diversity in forms of C4 in the genus Cleome (Cleomaceae). Ann.Bot. 107:269-283<br /> <br /> 27. Li, T., S. Huang, W. Z. Jiang, D. Wright and others. 2011. TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain. Nucleic Acids Research 39:359-372<br /> <br /> 28. Li, T., S. Huang, X. F. Zhao, D. A. Wright and others. 2011. Modularly assembled designer TAL effector nucleases for targeted gene knockout and gene replacement in eukaryotes. Nucleic Acids Research 39:6315-6325<br /> <br /> 29. Lin, P. C., M. C. Pomeranz, Y. Jikumaru, S. G. Kang and others. 2011. The Arabidopsis tandem zinc finger protein AtTZF1 affects ABA- and GA-mediated growth, stress and gene expression responses. Plant J. 65:253-268<br /> <br /> 30. Liu, X., F. Yu, and S. Rodermel. 2010. An Arabidopsis pentatricopeptide repeat protein, SUPPRESSOR OF VARIEGATION7, is required for FtsH-mediated chloroplast biogenesis. Plant Physiol 154:1588-1601<br /> <br /> 31. Liu, X., F. Yu, and S. Rodermel. 2010. Arabidopsis chloroplast FtsH, var2 and suppressors of var2 leaf variegation: a review. J.Integr.Plant Biol. 52:750-761<br /> <br /> 32. Liu, X., S. R. Rodermel, and F. Yu. 2010. A var2 leaf variegation suppressor locus, SUPPRESSOR OF VARIEGATION3, encodes a putative chloroplast translation elongation factor that is important for chloroplast development in the cold. BMC Plant Biol. 10:287<br /> <br /> 33. Ma, Y., S. V. Pollock, Y. Xiao, K. Cunnusamy and others. 2011. Identification of a novel gene, CIA6, required for normal pyrenoid formation in Chlamydomonas reinhardtii. Plant Physiol 156:884-896<br /> <br /> 34. McDonald, A. E., A. G. Ivanov, R. Bode, D. P. Maxwell and others. 2011. Flexibility in photosynthetic electron transport: the physiological role of plastoquinol terminal oxidase (PTOX). Biochim.Biophys.Acta 1807:954-967<br /> <br /> 35. McMaster, G., D. A. Edmunds, W. W. Wilhelm, D. C. Nielsen and others. 2011. PhenologyMMS: A program to simulate crop phenological responses to water stress. Computers and "Electron Agric 77:118-125<br /> <br /> 36. Moroney, J. V., Y. Ma, W. D. Frey, K. A. Fusilier and others. 2011. The carbonic anhydrase isoforms of Chlamydomonas reinhardtii: intracellular location, expression, and physiological roles. Photosynth.Res 109:133-149<br /> <br /> 37. Mutava, R. N., P. V. V. Prasad, M. R. Tuinstra, M. D. Kofoid and others. 2011. Characterization of sorghum genotypes for traits related to drought tolerance. Field Crops Research 123:10-18<br /> <br /> 38. Offermann, S., T. W. Okita, and G. E. Edwards. 2011. Resolving the compartmentation and function of C4 photosynthesis in the single-cell C4 species Bienertia sinuspersici. Plant Physiol 155:1612-1628<br /> <br /> 39. Offermann, S., T. W. Okita, and G. E. Edwards. 2011. How do single cell C4 species form dimorphic chloroplasts? Plant Signal Behav. 6:762-765<br /> <br /> 40. Oh, M. H., X. Wu, S. D. Clouse, and S. C. Huber. 2011. Functional importance of BAK1 tyrosine phosphorylation in vivo. Plant Signal.Behav. 6:400-405<br /> <br /> 41. Oh, M. H., J. Sun, D. H. Oh, R. E. Zielinski and others. 2011. Enhancing Arabidopsis leaf growth by engineering the BRASSINOSTEROID INSENSITIVE1 receptor kinase. Plant Physiol 157:120-131<br /> <br /> 42. Oliver, M. J., L. N. Guo, D. C. Alexander, J. A. Ryals and others. 2011. A sister group contrast using untargeted global metabolomic analysis delineates the biochemical regulation underlying desiccation tolerance in Sporobolus stapfianus. Plant Cell 23:1231-1248<br /> <br /> 43. Overstreet, L. F., G. D. Hoyt, and J. Imbriani. 2010. Comparing nematode and earthworm communities under combinations of conventional and conservation vegetable production practices. Soil & Tillage Research 110:42-50<br /> <br /> 44. Parry, M. A. J., M. Reynolds, M. E. Salvucci, C. Raines and others. 2011. Raising yield potential of wheat: Increasing photosynthetic capacity and efficiency. J.Exp.Bot. 62:453-467<br /> <br /> 45. Pelletreau, K. N., D. Bhattacharya, D. C. Price, J. M. Worful and others. 2011. Sea Slug Kleptoplasty and Plastid Maintenance in a Metazoan. Plant Physiology 155:1561-1565<br /> <br /> 46. Pengelly, J. L., S. Kwasny, S. Bala, J. R. Evans and others. 2011. Functional analysis of corn husk photosynthesis. Plant Physiol 156:503-513<br /> <br /> 47. Pomeranz, M., P. C. Lin, J. Finer, and J. C. Jang. 2010. AtTZF gene family localizes to cytoplasmic foci. Plant Signal.Behav. 5:190-192<br /> <br /> 48. Pomeranz, M., J. Finer, and J. C. Jang. 2011. Putative molecular mechanisms underlying tandem CCCH zinc finger protein mediated plant growth, stress, and gene expression responses. Plant Signal.Behav. 6:647-651<br /> <br /> 49. Pomeranz, M., L. Zhang, J. Finer, and J. C. Jang. 2011. Can AtTZF1 act as a transcriptional activator or repressor in plants? Plant Signal.Behav. 6:719-722<br /> <br /> 50. Pomeranz, M. C., C. Hah, P. C. Lin, S. G. Kang and others. 2010. The Arabidopsis tandem zinc finger protein AtTZF1 traffics between the nucleus and cytoplasmic foci and binds both DNA and RNA. Plant Physiol 152:151-165<br /> <br /> 51. Prasad, P. V. V., K. J. Boote, and L. H. Allen. 2011. Longevity and temperature response of pollen as affected by elevated growth temperature and carbon dioxide in peanut and grain sorghum. Environmental and Experimental Botany 70:51-57<br /> <br /> 52. Prasad, P. V. V., S. R. Pisipati, I. Momcilovic, and Z. Ristic. 2011. 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Impact Statements

1. Discovery of a new lipid remodeling enzyme at the chloroplast envelope and a new mechanism for plants to cope with freezing stress inform novel strategies for crop protection and yield increase.
2. Insight into the function of the PTOX and protein in photosynthesis, plant development and plant stress responses might lead to the design of strategies to manipulate the photosynthetic capacity.
3. Engineered changes in either the large or small subunit of RuBisCo far from the active site can influence carboxylation catalytic efficiency and CO2/O2 specificity. These regions may serve as targets for either the design of an improved RuBisCo.
4. Elucidation of the three-dimensional structure of Rubisco activase provides new insights into the biochemical regulation of CO2 assimilation.
5. Discovery of differences in the structure of photosynthetic cells, photorespiration, and water use efficiency between rice and wild relatives provides information which may be utilized for improving rice productivity.
6. Analyses of mRNA transcript changes that occur when Chlamydomonas cells are shifted from CO2 replete to CO2 depleted conditions have identified new physical components of the carbon concentrating mechanism (CCM) and regulatory factors of the process.
7. Understanding the mechanisms that allow Chlamydomonas to acclimate to low CO2 concentrations is important for evaluating the potential for increasing biomass yield from microalgae and the transfer of all or part of this CCM into higher plants.
8. Development of TAL Effector Nuclease (TALEN) technology has provided strong proof of concept demonstrating applicability to the engineering of algae and plants.
9. Understanding the conversion of photosynthetic sugars into triacylglycerols will be essential for the engineering of novel biofuel crops and algae.
10. Information on partitioning of photosynthates to cell-wall constituents will be potentially valuable for future bioenergy applications.
11. Research on altering the end product of photosynthesis will provide needed information to lift the limits on photosynthesis imposed by feedback effects and will show how sucrose/starch partitioning in photosynthesis affects plant development through sugar signaling.
12. Work on unique transcription factor protein family (TZF) with effects on plant growth, development, and stress response provides novel insights towards enhancement of biomass and stress tolerance.
13. Novel genes involved in sensing energy levels have proven useful in altering biomass through engineering.
14. Discovery of circadian-clock controlled regulatory patterns in metabolites in CAM plants will provide a comprehensive view of the metabolic flux required for the performance of CAM, informing efforts to introduce this water saving pathway into crops.
15. Diurnal measurement of photosynthesis in cotton plants revealed details about the onset of heat and drought stress that can be used in a selection strategy for identifying more stress tolerant plants.
16. A fundamental understanding of receptor kinase-mediated phosphorylation may provide new approaches to increase plant productivity and control response to environmental stress.
17. Studies of salt stress resistance genes show potential for major progress in developing salt tolerant crops for that quarter of the worlds agricultural crop lands that are salt-susceptible.
18. By elucidating the agents responsible for the continuous corn yield penalty, factors will be identified that can be managed and management recommendations to alleviate the yield penalty can be made.
19. Breeding and selection for improved plant performance under different stresses can be accelerated by the development of high-throughput methods for phenotyping plant responses.

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Impact Statements

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