Basil J. Nikolau - US grants

This research focuses on understanding the significance and role of biotin enzymes in plant metabolism. To achieve this goal the unique somatic embryogenesis system of carrot has been utilized to study these proteins. During the development of somatic embryos of carrot the steady state levels of one of the six biotin-containing polypeptides increases dramatically. This biotin protein has been purified and polyclonal antibodies against it have been prepared. Using these antibodies a cDNA clone coding for this biotin protein has been isolated. This novel biotin protein is being investigated through the utilization of these antibodies and the cDNA clone as specific probes to investigate this novel biotin protein. These investigations concentrate on elucidating the function and regulation of this biotin enzyme. Initial experiments with the antibodies have identified this biotin protein as representing an acetyl CoA carboxylase. The function of this form of acetyl-CoA carboxylase in plant metabolism is being elucidated by 1) identifying the subcellular and tissue distribution of this protein in the carrot plant and 2) elucidating the molecular genetic mechanisms that regulate the accumulation of this protein in response to developmental and environmental signals. Biotin is an essential component of a set of enzymes that have important metabolic functions. The structure, function and regulation of a number of biotin enzymes is extensively studied in a diverse set of organisms. However, very little is known about the biotin enzymes of plants. This research investigates the function and regulation of plant biotin- containing enzymes in order to provide a better understanding of their role in plant metabolism.***//

9316832 Schnable The composition of the cuticular wax of plants is complex and incompletely known, and little is known regarding the biosynthesis of this material(s). The glossy8 locus, one of the seventeen loci known to be involved in the biosynthesis of this wax in maize, has been molecularly cloned by tagging with a transposable element. In addition, the cer2 locus of Arabidopsis, which is involved in cuticular wax biosynthesis, has been localized to a 70-kb segment of the genome. The cer2 locus will be identified by genetic complementation tests. Both genes will then be used to study the biochemical and genetic mechanisms involved in the biosynthesis of these unique and biologically significant plant lipids. The cuticular waxes of plants constitute the first barrier to the environment and the stresses it imposes on plants. Thus, these waxes lay an important role in plant survival. The composition of these waxes and the biosynthetic pathways by which they are formed are incompletely understood. This project will make use of molecularly cloned genes to examine the gene products of the biosynthetic pathway and thus enlarge our understanding of it. ***

9507549 Wurtele The maintenance of a balance between anabolism and catabolism is crucial to the net biosynthesis of phytochemicals. This metabolic balance is particularly evident during seedling development when large quantities of seed reserve (lipids, proteins, and carbohydrates) are catabolized. The resultant intermediates are then utilized as substrates for the biosynthesis of new phytochemicals, or they are respired to generate ATP, which supports the biosynthetic reactions. These metabolic interconversions require the coordinate action and regulation of many antagonistic processes. This award will study the catabolism of leucine and subsequent utilization of the derived carbon in young seedlings of soybean and Arabidopsis, as an example of how organisms coordinate complex interconnected metabolic pathways to achieve net interconversions of biomolecules. In plants, the catabolism of leucine is thought to occur in mitochondria, via a set of reactions which are partly in common with a pathway by which mevalonate may be metabolized to non-isoprenoid compounds. Acetyl-CoA thus formed from leucine and mevalonate can be further metabolized for respiration, gluconeogenesis, and lipogenesis. These investigations will define previously unknown mitochondrial metabolic processes and begin to indicte how they interface with metabolism in other organelles. The understanding of catabolic provesses will ultimately provide a better understanding of how plants maintain a balance between anabolism and catabolism. Comprehension of how this balance is regulated may open new, as yet unforeseen, avenues to the improvement of agricultural production.

9808559 Schnable Survival requires that organisms produce a protective barrier between themselves and their environment. Because organisms must be able to sense and respond to environmental signals, this barrier must also allow for an exchange of molecules and information. In plants, the cuticle serves as this barrier. In addition to functioning as a water barrier, it has been suggested that the cuticle provides frost and UV resistance and plays a role in plant-pathogen interactions. Furthermore, the nature of the cuticle greatly affects the deposition and behavior of agricultural chemicals such as pesticides, growth regulators and foliar nutrients sprayed on plants. Cuticular waxes are an important component of the cuticle. They are complex mixtures of acyl derivatives of very long-chain fatty acids made in plant epidermal cells by VLCFA elongase enzyme systems. A maize gene (gl8) responsible for the production of one of these enzymatic functions has been cloned, and the other genes required for the production of functional VLCFA elongase enzyme systems will be cloned. This integrated research approach will enhance our understanding of the mechanism by which the elongation of acyl chains occurs. Because similar types elongation reactions are important in the biosynthesis of many classes of molecules (including polyketides, flavonoids, and stilbenoids) which function as antibiotics, plant-pathogen toxins, pigments and protective compounds, the proposed studies will impact our understanding and ability to manipulate many fundamental biological processes

An organism's ability to maintain a balance between anabolism and catabolism is crucial to achieving net accumulation of biomolecules. In addition, this metabolic balance must be integrated with the rest of the organism's metabolism and remain flexible to respond to a wide variety of developmental and environmental signals. For stationary organisms such as plants, accommodation to environmental parameters is particularly critical. Research on the metabolic function of methylcrotonyl-CoA carboxylase (MCCase) indicates that plant leucine catabolism is networked with other metabolic and physiological processes. Hence, the study of plant leucine catabolism may offer novel insights on how organisms network metabolism to achieve several levels of regulation of complex reticulated processes. This project focuses on the regulation of leucine catabolism in Arabidopsis as an example of how complex interconnected metabolic pathways are controlled to achieve net interconversions of biomolecules. It has been shown that plants catabolize leucine via a mitochondrial pathway that requires the enzyme MCCase. In addition, characterizations of MCCase-deficient genetic stocks indicate that leucine catabolism in plants is shared between mitochondria and peroxisomes. The aim of this project is to elucidate the signaling and regulatory mechanisms that allocate leucine to the mitochondrial versus the peroxisomal catabolic pathways, and to determine how these catabolic pathways are networked with other metabolic processes. To do this, three specific hypotheses will be tested: Hypothesis 1: A block in mitochondrial leucine catabolism is compensated by global shifts in patterns of gene expression. Hypothesis 2: MCCase is the point of integration of leucine catabolism and isoprenoid metabolism. Hypothesis 3: Mitochondrial leucine catabolism is regulated by the metabolic status of the organism in part via changes in MCCase gene transcription.
Plants take in simple molecules such as carbon dioxide, and water. Using energy from the sun, these are converted to complex compounds such as starch, proteins, fibers, oils and pharmaceuticals. In addition, these complex compounds are constantly broken down to provide energy for growth as well as to provide the new compounds needed by each plant cell to respond to an ever-changing environment. The net accumulation of useful biomolecules by the plant is thus dependent on the balance between two processes. The long-term goal of this research is to exploit the synergy inherent in a combined genetic and biochemical approach that will ultimately reveal how plants maintain such a balance.

Partial support is requested for the 3rd International Congress on Plant Metabolomics to be held from June 3 to 6, 2004 at Iowa State University, Ames, Iowa. The meeting will include presentations by 9 invited speakers who are leaders in different aspects of plant metabolomics. Up to twelve additional speakers will be selected from submitted abstracts for short oral presentations. The meeting also includes poster sessions and time for informal discussions. The conference incorporates programmatic and educational features to encourage meaningful participation by scientists and students representing diversity in experimental system, career level, ethnicity and gender.


Intellectual Merit: Since the sequencing of the first plant genome (Arabidopsis), it has become possible to envision the functionality of the entire complement of genes that encompasses a plant. This challenge has manifested a systems-based approach to visualize the expression of the entire genome. Specifically, technologies for globally measuring gene expression at the mRNA (transcriptomics) and protein (proteomics) levels have been developed. Metabolomics is the technology for globally measuring gene expression at the metabolite level, which probably represents the ultimate expression of the functionality of a genome that can be chemically defined. Metabolomics is an emerging field. The International Congress on Plant Metabolomics was initiated just 2 years ago and is the medium that brings together the leading practitioners in this new emerging area of plant functional genomics. Previous meetings have been held in Europe, and this, 3rd meeting in this series will be the first to be held in the US. The Congress will for the first time in this country provide a landmark opportunity for plant biologists, chemists and bioinformaticists to meet, discuss and interact in the context of a major international meeting on plant metabolomics. The Symposium will be one of an ongoing series of symposia sponsored by the Plant Sciences Institute at Iowa State University. The program will address the current and potential applications of metabolomics in gene discovery and directed genome modifications.

Broader Impacts: This symposium is designed to initiate and foster interactions among practitioners of metabolomics and researchers studying plant biology. The speakers have been chosen to present investigations on a variety of related topics, including the development of new analytical technologies, statistical analysis of metabolomics data, and the role of metabolomics in solving biological problems via a systems approach. Approximately one third of the invited speakers are women; up to twelve additional short talks will be selected from submitted abstracts to increase diversity in career level and to enhance participation of underrepresented groups. An important purpose of these symposia is to promote interactions and new collaborations among researchers who might not normally come together at a single symposium. In addition to the attendance of graduate students and post-doctoral researchers, special efforts will be made to encourage participation by NSF-REUs, other undergraduate students, and interested high school teachers. Long-standing relationships of ISU with historically black institutions and undergraduate research experience programs will foster attendance of underrepresented students.

Very long chain fatty acids (VLCFAs), those that contain 20 or more carbon units, are incorporated into a wide variety of physiologically significant phytochemicals, including cuticular waxes, cutin, suberin, sphingolipids, and some phospholipids and seed oils. Genes that encode several of the enzymes involved in the production of VLCFAs have been isolated. Based on the finding that maize mutants that block the production of one of these enzymes are embryo lethal, VLCFAs must be essential during maize embryogenesis. Mutants that block the accumulation of essential VLCFAs therefore provide an ideal experimental system to determine the essential role of VLCFA-derived phytochemicals in plant development.
The proposed project will define the essential developmental role(s) of VLCFAs, by determining the morphological differences that distinguish the development of mutant embryos from normal embryos. It will also establish which VLCFA-derived compound(s) are essential during embryo development, by comparing the accumulation of VLCFA-derived compounds in normal and mutant embryos. Finally, mutants that overcome the embryo lethality associated with genetic blocks in the production of VLCFAs will be isolated. In the long term, these mutants can be used to dissect the diverse network of biochemical pathways that utilize VLCFAs.
The proposed studies meet the challenge of understanding the complex interplay between biochemistry and development by continuing to exploit the synergy inherent in a long-term research collaboration that integrates genetic and biochemical approaches. This project will provide excellent cross-disciplinary training experiences to a diverse group of early-career scientists because it will use a combination of hypothesis-driven and genome-wide analyses, and apply cutting-edge technologies to address significant questions in plant biology that have proven recalcitrant to a mono-disciplinary approach.

Intellectual merit: Biotin is an essential molecule, whose primary function has been ascribed as a catalytic cofactor required by enzymes involved in diverse metabolic processes. Recent studies indicate that biotin is also a regulatory molecule that appears to play critical roles in controlling transcriptional and post-transcriptional mechanisms in gene expression. This research will identify and characterize the biochemical and physiological functions of genes associated with the biotin metabolic network of eukaryotic organisms, using Arabidopsis as a model. Plants are ideally suited for these studies as they, along with some microbes, are the primary organisms that can synthesize this molecule de novo; all other organisms must acquire this molecule from their diets or from the environment. This project will characterize aspects of the biotin metabolic network that are unique to the organism under study, namely Arabidopsis, as a model plant. Specifically: 1) how do developmental and environmental cues regulate biotin biosynthesis; these studies will take advantage of the two biotin biosynthetic genes that have recently been identified by the investigators, BIO1 and BIO2; and 2) what is the biochemical and physiological function of plant-specific biotin-containing proteins. These are methylcrotonyl-CoA carboxylase (encoded by genes At1g03090 and At4g34030), the seed-specific biotin protein (encoded by gene At2g42560), and three biotin carrier-like proteins (encoded by genes At1g52670, At3g15690, At3g56130); and 3) what are the mechanisms by which biotin regulates gene expression at both the transcriptional and post-transcriptional level.
Broader impact: The project will be a vehicle for the education and training of a new cadre of scientists at the undergraduate, graduate and post-graduate levels. In-lab research experiences will be provided for exposing young scientists to multidisciplinary basic research in functional genomics and biotechnology. The research will define the organization of a complex metabolic network that is structured around the catalytic and regulatory functions of biotin. This achievement will provide novel insights into how such complex networks are regulated. The goal is to develop an example of how to view metabolism as a complex integrated network rather than the classical textbook model of a set of linear pathways. The development of this view of metabolism is possible only in the context of an organism whose genome is completely sequenced. Such an understanding of metabolism is required in order to comprehensively understand how metabolic processes are interactively regulated by developmental and environmental cues.

The functions of over 1/3 of the annotated protein-coding genes of the Arabidopsis genome are still unknown, and the annotation of an even larger portion of the genome is not sufficiently accurate for unambiguous assignment of function at the biochemical and physiological levels. This project will bring together a consortium of multidisciplinary collaborators to establish pipelines for generating metabolomics data-streams and to provide statistical and computational interpretation of the resulting integrated datasets. The goal is to develop metabolite-profiling capabilities that will enhance the research community's ability to formulate testable hypotheses concerning Arabidopsis gene functions. The consortium has developed metabolomic platforms that together detect approximately 1,800 metabolites, of which 900 are chemically defined. The aim of the project is to apply these established metabolite-profiling platforms to reveal changes in the metabolome associated with knockout mutations in up to 200 Arabidopsis genes of unknown function and compare these to similar mutants in 50 genes of known function. The consortium will disseminate these data via the existing multi-functional metabolomics database: www.plantmetabolomics.org. Enhancement of this database and associated statistical and visualization toolsets will enable researchers to formulate testable computational models of the metabolic network of Arabidopsis. The successful completion of these goals and integration with other NSF-sponsored functional genomics and cyber infrastructure developments will generate transformational resources for ultimately modeling the complex metabolism of Arabidopsis.

Broader Impacts
The project will develop new resources for the research community that will enhance the capability to globally profile genome expression at the metabolite level. These metabolite resources, in collaboration with other NSF-funded resource development projects, will enable researchers in the community to formulate credible, testable hypotheses concerning gene function. The project will foster the development of the science of metabolomics as a functional genomics tool through workshops, internships and organization of national and international meetings. The project will also develop new activities to enhance the impact of science education and training in the community, by conducting workshops for researchers at consortium labs and at international biological meetings. In addition, research internships will be offered to undergraduate students, eight of whom will have the opportunity to experience international science training in a European genomics laboratory. These research-based training internships will illustrate to the students the synergy that accompanies the integrated applications of chemistry, biochemistry, genetics, bioinformatics and computational sciences to solving complex biological problems.

Intellectual Merit:
Alkamides (amides linked to fatty acids) are novel plant secondary compounds that are poorly characterized, but that accumulate to significant levels in Echinacea, making this plant of choice to study these compounds and their biosynthesis. Our understanding of alkamide function in plants is in its infancy, but recent data suggest these compounds may have novel signaling functions in addition to insecticidal properties. This work will lay the foundation for further studies of the importance of these compounds in plant biology as well as open up research on this plant genus which has been used medicinally for hundreds of years. Because the pathways in alkamide biosynthesis involve amino acid metabolism and novel chemical linkages, there are also multiple future biotechnological applications for the enzymes involved. The proposed research will strategically apply high-throughput global profiling technologies to elucidate this natural product biosynthetic pathway. Alkamides appear to be biologically assembled via a modular metabolic pathway that may be an adaptation of amino acid and fatty acid metabolism. Experiments that combine metabolite profiling, metabolic flux studies and transcriptomics will be used to identify genes and enzymes that assemble a diverse collage of alkamides. Defining the alkamide pathway offers the potential of discovering new metabolic processes that generate novel combinations of chemical functionalities, which have wide-ranging applications (e.g., lubrication and detergent industries). In addition, this research project outlines a general methodology that should be broadly applicable to discovering how primary and specialized plant metabolism are juxtaposed and evolve to generate the physiochemical phenotypic differences among plant taxonomic groups. The multilayered bio-prospecting to be used offers the opportunity to browse the metabolic repertoire of an organism, and the system-wide knowledge of the involved biochemical processes should translate to the creation of novel bio-derived compounds relevant to the chemical industries, as well as strategies for pest resistance.

Broader Impacts:
This project will facilitate a multidisciplinary partnership between IUPUI, ISU researchers and scientists at the USDA North Central Regional Plant Introduction Station. The collaboration established to conduct this research will mentor young scientists at the increasingly uncommon intersection of organic chemistry, mechanistic biochemistry and functional genomics through the coeducation of undergraduates, graduate students and post-doctoral fellows. Students at IUPUI, a large, urban university, will be drawn from a trainee pool rich in underrepresented minority and first generation students in the McNair and Diversity Scholars STEM research programs. These interactions will allow the project to operate synergistically with IUPUI campus initiatives to mentor undergraduate students toward graduate education. As an outreach activity specific to IUPUI, high school teachers will experience hands-on research. In conjunction with the graduate students, they will engage in experiments leading to biochemical lessons suitable for use at their home schools. This effort dovetails with ongoing NSF GK-12 participation in IUPUI's research programs. At ISU, undergraduates will translate the proposed research into a module in a new course in biotechnological biochemistry.

This Major Research Instrumentation (MRI) award funds the acquisition of a Bruker-Daltonics SolariXHybrid FTMS Bundle mass spectrometer system at the Iowa State University WM Keck Metabolomics Research Laboratory, and empowers multidisciplinary research from a wide sweep of departments that integrate disciplines of chemistry and biochemistry, biological sciences and engineering. The new instrument system will provide researchers with unprecedented data concerning biological molecules and thus enhance the research infrastructure of the University and the State. The projects supported by the new instrument system attempt to meet the challenge of deciphering functional understanding of genome expression at the metabolic level, and are a mixture of hypothesis-driven research on the structure and regulation of specific metabolic and gene networks, and high-throughput global profiling projects that seek to generate hypotheses concerning gene functionalities. The impact of these research projects will be greatly enhanced the SolariXHybrid FTMS Bundle mass spectrometer system because researchers will be able to more rigorously determine the chemical identities of metabolites and hence generate and test more robust hypotheses concerning the metabolic functionalities of gene products and gene networks.

In addition, this instrument will provide faculty a vehicle for the multi-disciplinary research-based training of students, who will gain insights into how to conduct modern metabolic studies, integrating advances in chemistry and genomics biology. Post-docs, graduate students, undergraduate students from Iowa State University and neighboring colleges, and high-school students and teachers from the State of Iowa will also participate in research. These latter outreach activities will be coordinated through summer workshops, internship programs and NSF-REU programs that are already in place through the Centers (e.g., CBiRC) and projects (e.g., The Arabidopsis 2010 Metabolomics Consortium Project) that will utilize the instrument. Therefore students will be broadly mentored and trained to meet the emerging technological challenges of the 21st century. Results from the research projects will be disseminated through student and faculty presentations at regional or national meetings, and through publications in the peer-reviewed literature.

Abstract

PI Name: Jacqueline Shanks
Institution: Iowa State University
Proposal Number: 0938157

EFRI: EFRI-HyBi: Bioengineering a system for the direct production of
biological hydrocarbons for biofuels

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)

Intellectual merit: This project will develop new bio-engineering technology for transforming the current liquid fuel industry from using fossil-carbon feedstocks to using biorenewable feedstocks that are at the chemical level identical to gasoline and diesel fuels, namely biologically-generated hydrocarbons. The engineering system envisioned is a photosynthetic-based organism that will have the bio engineered ability to chemically-reduce atmospheric CO2 to simple hydrocarbons (e.g., n-alkanes and n-alkenes), using sunlight as the source of renewable energy. Such metabolic conversions are known to occur in discreet places in the biosphere, e.g., the epidermis of plants and insects, and as a carbon/energy-storage mechanism by certain algae. The goal is to conduct multidisciplinary studies that will identify the mechanisms and genetic elements that encode the biocatalyst(s) that generate these hydrocarbons in biological systems. The PIs will explore the use these isolated genetic elements to establish to bio-engineer crops or bioengineer photosynthetic microbes as the production platform to realize the vision of producing a biological hydrocarbon based fuel. The proposed research will for the first time lead to fundamental knowledge concerning the structure and mechanism of the biocatalyst that generates biological hydrocarbons. And, the efficient use of this novel biocatalyst in a production biological host will require the optimization of bioengineering principles so as to proficiently integrate the biocatalyst into a pre-existing metabolic network without compromising the biological competence of the host. These later optimizations will integrate concepts of biological control principles with engineering proficiencies.

Broader impacts: This project brings together a collaborative of biologists and engineers to demonstrate a paradigm of how fundamental molecular biological research can be integrated with disciplines of engineering to generate new bio-engineered organisms that can be used as a sustainable production platform to meet the global demands for new liquid biofuels. This multidisciplinary team-approach will be an ideal venue for training and educating new cadre of students and researchers of the synergy that is gained from the novel combination of biological and engineering research. This type of training, which integrates biological, genetic and biochemical research with quantitative engineering ad economic perspectives is increasingly important to meet the grand challenges of the 21st century, in which increased global population demands will tax the natural system's ability to maintain a sustainable human societal infrastructure. In this context, the PIs will foster an international collaborative (the International BioHydrocarbon Group) that will be a venue for trainee exchanges, which will provide students and post-docs an international perspective in their research-based training. In addition, they will incorporate an REU program that will recruit undergraduate students from institutions that serve primarily under-represented groups, and foster their scientific and engineering development through research-based education. Thus, the PIs will be able to exhibit within this project that meeting an emerging global challenge will demand new intellectual perspectives and solutions from the brightest young engineers and scientists, and these will need to be developed in an increasingly globally integrated manner. Hence, the dissemination of the combined research outcomes generated by the biological and engineering students and post-docs of the project will impact the respective training and education communities by setting precedence for optimally training and mentoring young investigators and thus transforming an emerging field.

TITLE: New GK-12: Growing the green collar workforce for the 21st century
INSTITUTION: Iowa State University

This project will support graduate student fellows who will act as the human interface to motivationally communicate to middle school teachers and students the excitement and innovations that are the scientific underpinnings of today?s technologically savvy, modern society. The rationale for this program is the need to solidly ground this country's educators and young students in STEM-theme pedagogy. In the long-term such research-based education and training is a prerequisite for facing the emerging societal challenge to generate technologies that will maintain a sustainable standard of living, which ultimately will be dependent on renewable sources of energy and industrial materials. This program builds on Iowa State University's established scholarly infrastructures that are designed to be interdisciplinary and address key technology gaps for harnessing renewable energy sources (solar energy) and generate carbon-based energy and chemicals using atmospheric CO2 as the ultimate carbon-source. This interdisciplinary research integrates core themes that are grounded in STEM research, which include the fields of plant sciences, agro-engineering, biocatalysis, biochemical pathway discovery and metabolic engineering, bioinformatics, biomaterials, chemical catalysis, and mathematics. Broader impacts of the project include increased professional development opportunities for middle school science teachers; expanded educational and career opportunities for K12 students in STEM disciplines, especially those from underrepresented minorities and economically disadvantaged; benefits to US economy resulting from well-trained, globally competitive workforce; and recruitment of female and underrepresented minority graduate students to associated academic programs.

This Partnerships for Innovation project from Iowa State University (ISU) will accelerate fundamental understanding in plant science and breeding with a screening platform that significantly reduces the time and costs for selecting and developing new elite plant varieties. The immediate goal is to build upon and re-engineer a promising near infrared (NIR)-based seed-screening platform to increase its performance and accuracy and broaden its applications. The envisioned platform is based on two complementary components: a) NIR molecular spectroscopy and chemometrics integrated with automated, nondestructive high-throughput screening of seeds and other plant materials, and b) customized development of application methodologies that screen extant plant populations based on biochemical phenotypes. This project seeks to meet the challenge presented by the success of high-throughput DNA sequencing and analysis of plant genomes, by establishing analogous high-throughput technologies to analyze a plant's biochemical phenotype. This capability will enhance the ability of plant scientists and plant breeders to utilize genomics and develop improved new cultivars needed to accommodate the global population explosion anticipated in the 21st century. The proposed phenotype-screening platform will have a significant impact on food, feed, and bio-energy production systems because, for the first time, plant scientists and breeders will have access to a unique research tool that will routinely assess, at low cost, multiple plant traits simultaneously and non-destructively based upon biochemical phenotypes.

The broader impacts of this research are societal, economic, and learning related. At the societal level, plants have a remarkable, multi-faceted impact on economies, jobs, health, fuel, climate change/carbon sequestration, food security, world peace, the environment, and our overall well being. Better understanding of plant biology, improving existing varieties, and developing new ones are all key research areas where biochemical phenotype screening will produce invaluable data for achieving sustainable and nurturing ecosystems for a rapidly changing planet. At the economic level, these improvements will greatly enhance the versatility of the screening platform, expand the user base, and make lower cost screening accessible to more public and commercial plant scientists and breeders. At the learning level, the KEP participants will learn from each other. The project will be a vehicle for Iowa State University faculty and students and small businesses to understand how each other's abilities can be integrated to enable technological solutions to real world-challenges.

Partners at the inception of the project are Brownseed Genetics (Bay City, WI), Genetic Enterprises International (Johnston, IA), Kemin Industries (Des Moines, IA), MTEC BioAnalytics (Ames, IA), Schillinger Genetics (West Des Moines, IA), and Sustainable Oils (Bozeman, MT). Five ISU researchers and these six small businesses will collaborate in a Knowledge Enhancing Partnership (KEP). MTEC BioAnalytics will bring NIR seed screening technology expertise to the project while the other companies have expertise in seed and other plant based products. MTEC BioAnalytics will provide key highthroughput screening platform components (e.g., ultra fast fiber optic switch), engineering design, and the others will provide the use of proprietary and unique collections of characterized seeds that express variation in desirable biochemical phenotypes.

This Innovation Corps project will explore the potential of developing a marketable innovation based on the use of 3-ketoacyl ACP synthase III (KASIII) enzymes. KASIII catalyzes the first carbon-carbon forming reaction in fatty acid biosynthesis, and thus determines the chemical nature of the omega of the fatty acids produced.

The proposed innovation is based on engineering of of the biocatalyst, KASIII, and would result in the production of bi-functional molecules that can act as the monomer percusors for making bio-based plastics. This innovation will provide novel bi-functional, bio-based feed stocks to the emerging bio-renewable chemical industry for the production of novel "green" plastics and specialty chemicals. The proposed I-Corps team envisions a technology that will be initially targeted to the synthesis of hydroxy-fatty acids, but can be readily advanced upon to produce other bi-functional monomers (e.g., amino-fatty acids, which can be used to produce polyamides). If successful, this technology will lead to chemical products that can act as substitutes for petroleum-based chemical products, and be precursors for novel bio-based products.

The aerial surfaces of land plants are protected by unique lipids, which provide a primary line of defense against numerous biological and environmental stresses. The surface lipids on the stigmatic silks of maize are biologically unique because they are rich in hydrocarbons, which are the inert end-point metabolites of the surface lipid network. This project will utilize maize silks as the model biological system to provide a fundamental understanding of this unique, discrete metabolic process by comprehensively dissecting within a single organism the metabolic and genetic networks that produce important surface lipids. Moreover, this project will elucidate the specific surface lipid constituents of maize silks that provide critical protection against environmental stresses (water stress and insect feeding) that commonly impact crops like maize during the often-stressful period of pollination. All metabolite and transcriptome data will be deposited in the Plant Metabolomics Resource database (metnetdb.org/PMR) and the latter will also be deposited in the NCBI-SRA and NCBI-GEO databases. Quantitative trait locus (QTL) data will be searchable at MaizeGDB (http://maizegdb.org/qtl.php) and made available at the integrated web portal for QTL dissection, GeneNetwork (genenetwork.org). Seed stocks for quantitative genetic mapping populations will be deposited at the Maize Genetics Cooperative Stock Center (http://maizecoop.cropsci.uiuc.edu).
A significant societal impact of this project will be enhancement of the sustainability of US crop production via increased crop yields and/or decreased inputs. The identification of the protective surface lipids and the genes that produce them will provide the technological know how for applied breeding of customized lipid compositions that protect against many stresses, both in corn as well as in other crops. Further, the chemical similarity between silk surface lipids and petroleum components will be leveraged for applications in network bioengineering to produce advanced biofuels in other biological systems. During the project, >400 high school and college students from diverse socioeconomic, geographic and ethnic backgrounds will be exposed to hypothesis-driven research via three synergistic programs: A) a collaboration with Iowa State University's SCIENCE BOUND program will engage diverse secondary school students in scientific education via hands-on research modules; B) two SCIENCE BOUND students will conduct summer-research projects together with a high school teacher from a rural or high needs district and with a student from Chowan University, a 4-year undergrad institution with significant minority enrollment. These research experiences will help the teachers better engage an even larger number of students in scientific inquiry; and C) a discussion-based undergraduate course will be developed to serve as a bridge between fundamental knowledge learned through coursework and the application of that knowledge for conducting scientific research.

Roots interact with and respond to the biotic and abiotic environment in which they live (the rhizosphere) by qualitatively and quantitatively modulating material exuded by roots. These exudates use the language of chemistry to communicate between and among the biotic and abiotic components. Powerful DNA sequencing platforms and analytical tools for identifying chemical components are now available for profiling these interactions among the rhizosphere and the root components. The integrated application of these analytical strategies is limited by the ability to access and isolate exudates from roots and the rhizosphere. Existing exudate sampling tools are bulky, require large amounts of soil, and significantly alter the soil structure. This difficulty of sampling exudates has slowed the process of linking plant genetic determinants to rhizosphere microbiome genomic and metabolic features. This project addresses key design, fabrication, integration, and operation problems faced in developing next-generation root exudate sampling tools. The research will develop greatly-needed tools for probing the chemical exchange between plants and the micro- and macro-organisms in the rhizosphere. The root exudates are critical drivers of microbiome assembly and plant-pest/pathogen outcomes. The dynamic and environmentally responsive nature of root exudates illustrates the importance of developing sampling tools that are functional in a real-world situation, rather than the current tools that are limited to use in primarily artificial hydroponic and polymer-embedded systems. The samplers that will be developed will significantly impact the pace of research on rhizosphere microbiome by enabling continuous, spatially-resolved sampling of the microbes and exudates on roots grown in real-world conditions. This enhanced capability will meet societal needs to increase agricultural productivity for an increasing global population in the face of the uncertainties associated with climate-change, and thus develop new strategies to impact gains in agricultural productivity. This research will enhance interdisciplinary STEM workforce development by hosting at least two under-represented students in an undergraduate Howard Hughes Medical Institute summer internship program, providing research opportunities to four undergraduate senior students, and providing hands-on workshops to a high school Science Bound program to engage students in tech-transfer endeavors, while highlighting plant-microbe contributions to agriculture and global food security.

This project will elaborate advanced technology for gathering high spatiotemporal resolution data of metabolites and microbes in the rhizosphere. This objective will be met by developing a modular toolkit for the localized sampling of rhizosphere exudates from roots grown in soil matrices. This toolkit will consist of (i) a single site exudate sampler, which will serve as a building block of more complex modular sampling systems; (ii) distributed exudate samplers able to extract exudates from key locations with high spatial resolution; (iii) spine-like flexible exudate samplers, providing conformational fitting at the root-soil interface, which will maximize sampling at this crucial interface; and (iv) parallel gradient samplers positioned radially outward from a root, providing access to radial gradients of exudates. These samplers will be uniquely coupled with microfluidic sorters to enable automated separation and isolation of microbes from the collected exudates for simultaneous analysis of both the microbes and the soluble exudates. Furthermore, these samplers will integrate miniature tensiometers, which will allow monitoring of local soil potential condition at the sampling sites, and trigger the automatic start of sampling. Rendering such a "smart" device will improve temporal resolution of sampling. Validating the utility of these integrated devices will involve installing them, collecting and sorting samples, and analyzing the interactions between the rhizosphere and genetically specified maize roots, grown under gnotobiotic conditions with and without microbes. The multidisciplinary research has drawn expertise ranging from microsystems design and construction, microbiome and metabolomics, to address the proposed goal and deliver on the specific aims.