Mark J Guiltinan - US grants

EmBP-1 is a wheat basic-leucine zipper (bZIP) class transcription factor which has been implicated in the control of gene expression. Proposed experimentation will address the structure-function relationships of the DNA binding domain of EmBP-1. By analyzing mutant DNA binding proteins with altered binding preferences, insight can be gained of the ways in which amino acid sequences within the DNA binding domains of bZIP proteins encode functional specificities. The minimal domains for the dimerization and DNA binding functions of EmBP-1 will be determined using deletion analysis and in vitro binding assays. cDNA libraries encoding randomly mutagenized basic region domains will be constructed and probed with degenerate oligonucleotides to determine how changes in the amino acid sequences of the bZIP domains are manifested in altered binding specificities. The functional significance of these findings will be tested in vivo by expressing mutagenized versions of EmBP-1 in transgenic Arabidopsis plants. Additional, to examine the natural variation in bZIP sequences, members of the EmBP-1 gene family will be isolated from Arabidopsis. Because the leucine zipper class of transcription factors has been conserved from plants to animals, the knowledge gained from the proposed research will be applicable to the basic principles operating in most eukaryotic cells. %%% EmBP-1 is a wheat DNA binding protein involved in controlling gene expression. Understanding the way in which DNA binding proteins recognize specific DNA sequences will lead to a greater understanding of the mechanisms by which cells activate gene expression. This has great significance in understanding normal growth and development as well as in understanding aberrant gene expression which is often associated with human disease such as cancer. By analyzing mutant DNA binding proteins with altered binding preferences, insight can be gained of the ways in which this class of proteins function. Randomly mutagenizeds Dna binding domains will be constructed analyzed to determine how changes in the amino acid sequences are manifested in altered binding specificity. These findings will be tested by expressing mutagenized versions of EnBP-1 in genetically engineered plants, Additionally, to examine the natural variation in DNA binding sequences, related genes will be isolated from Arabidopsis (a model plant species used in molecular genetic studies). Because this type of protein is found within organisms ranging from plants to animals, the knowledge gained from the proposed research will be applicable to the basic principles operating in most cells.

9313436 Guiltinan This award allows Pennsylvania State University to renovate their complex of six greenhouses, a total of about 28,500 square feet of research and research training space. These greenhouses were built in 1949 and have not undergone any renovation since that time. The types of research carried out in this space include control of plant gene expression, control of plant cell expansion, biosynthesis of starch, plant growth regulation, effects of air pollutants on plants, and the molecular basis of cell recognition in plants. These greenhouses are also used for various graduate and undergraduate courses in plant science and specifically an interdisciplinary training program in advanced root biology. These facilities are used by 20 faculty, 8 postdoctoral fellows, 60 graduate students and about 145 undergraduate students. Thus, the renovations will impact a large number of mature researchers and existing research projects but a larger number of students being trained in biotechnology issues concerning plant science. This modernization project is important because of it's potential regional and national impact on the growing biotechnology infrastructure. ***

Abstract: Mark Guiltinan (#0215923)

A grant has been awarded to Pennsylvania State University under the direction of Dr. Mark Guiltinan to develop a Plant Growth Facility consisting of six growth chambers and a small greenhouse. The facility center will provide a physical home for the graduate programs, and an organizing role for the enhancement of Plant Sciences research, teaching and outreach programs.
The objectives of the center will be to: provide state-of-the-art research facilities for a core group of resident plant scientists, including current faculty and new hires; provide collaborative space in the new building to members of the Plant Science Center; develop and enhance collaborative interactions among plant scientists and with researchers
in other fields, at Penn State and elsewhere; improve the Plant Physiology and Ecological and Molecular Plant Physiology graduate education programs, and; develop outreach programs for the general public and for pre-college students.

This center will support research in plant sciences for diverse studies, but will be particularly important to the growing number of plant scientists using the model plant species, Arabidopsis thaliana. The completion of the Arabidopsis genome sequence, along with major new funding programs in plant genomics at the NSF and USDA,
have brought us to a new era in plant research, requiring high throughput facilities for functional analysis of genes.

Penn State has made a priority effort to enhance the life sciences; enhancement of the Plant Sciences Center will help us move forward on this path, by providing necessary facilities and maximizing laboratory space in the main building. This will in turn help to attract top faculty candidates who will be concerned about availability of such facilities. The facility will provide for the first time, an interdisciplinary, inter-college facility for high quality environmental growth of plants to support the many plant scientists at Penn State. It will also serve our teaching and outreach programs, providing a facility available to all for the growth of plants.

This NSF award by the Biotechnology, Biochemical and Biomass Engineering program supports work to develop low-cost bioreactor systems that will enhance the ability to utilize tissue culture to improve agricultural plant productivity. In addition to the typical paradigm of optimizing the chemical and environmental conditions within the bioreactor, this research seeks to demonstrate the ability to control plant development using transcriptional factors that control the expression of multiple genes. These transcriptional factors will be delivered to the plant tissues using Agrobacterium, a bacteria that has been developed to transiently deliver DNA to cultured plant tissues (developed with previous NSF support; BCS-0003926). The genes associated with plant embryo formation will be identified by examining patterns of gene expression during somatic embryogenesis. The concept of inducing embryo formation will be implemented using Cacao, the source of chocolate, due to its commercial (and social) value. In addition to providing an instructional case study that demonstrates the use of interdisciplinary principles of bioreactor design and molecular biology, the effort will produce thousands of fungal resistant plants that will be distributed to smallholder farmers in South America. In addition, the low-cost bioreactor technology has potential applications to a broad range of food and biomass crops.

1126373
Ferry

Mass spectrometry (MS) is one of the leading analytical tools exploited by the metabolomics community. Despite technological advances, it remains impossible for a single piece of MS instrumentation to provide all of the information sought in global analyses. As such, all of the centers of excellence in metabolomics utilize pipelines that exploit multiple analytical platforms (e.g., LC-MS, GCEI-MS, GC-CI-MS). The proposed AB Sciex TripleTOFTM 5600 is a high resolution, exact mass MS/MS platform that is specifically designed to provide the resolving power sought by the metabolomics community. This instrument provides a mass resolution of up to 40,000 full width at half height (FWHH) in both positive and negative ion modes, exact mass (1 ppm RMS) with a dynamic range of up to five orders of magnitude, a rapid acquisition rate (20 Hz) that fully exploits the resolving power of UPLC, and absolute sensitivity more usually associated with triple quadrupole instruments operating in Selected Reaction Monitoring (SRM) mode. In short, this instrument offers speed, accuracy and versatility to explore complex samples from a large user group with diverse needs.

The 20th Penn State plant Biology Symposium will focus on the important topic of "Plant Stress-Omics in a Changing Climate". Plenary sessions are focused on water and salinity, atmosphere (temperature, CO2, ozone), biotic/abiotic interactions and the use of novel tools. The conference is held at Penn State University, University Park takes place May 13-16, 2015. There will be two formal poster and two short talk sessions to feature research from talented junior scientists. The conference includes an impressive cadre of US and international speakers. The organizing committee and speaker list seems balanced in terms of broadening participation.

The workshop addresses an important topic in Plant Biology, namely how plants deal with increasing environmental stresses. Particularly noteworthy is a planned workshop preceding the meeting to provide training in novel mass spectrometric methods. These methods are necessary to determine metabolomics changes in response to various environmental stresses. The meeting is an ideal forum for students and postdocs to identify future mentors, and for established scientists to forge meaningful collaborations.

The human population is projected to grow to 9.6 billion by 2050, requiring a 50% increase in food and fiber production. A major constraint to this goal is crop losses due to microbial plant diseases which destroy approximately 15% of the world's total crop production every year. Advances in the science of plant disease control are needed to reduce crop losses to disease. Plant genomes encode thousands of genes involved in disease resistance, called the plant immune system. For some crops, modern science has made rapid progress in identifying disease resistance genes and using them in plant breeding, but for others such as long-lived tree crops, it is much more difficult to make rapid progress. This project will explore the plant immune system via a comprehensive study of the genes important for disease resistance to key pathogens of an important crop, cacao, which is the source of chocolate, and an important cash crop for millions of farmers in developing countries. The methods, tools and knowledge gained will be directly applicable to discovery of genes underlying important traits in other crops, especially trees and many perennial grasses. In addition to contributing to building a global partnership in reaching the goals of feeding a growing population sustainably, this project will involve students and young scientists in the US and in developing countries though international exchanges and collaborations.

This project will establish a new approach for use with perennial crop plants to identify candidate loci for disease resistance using a model tree crop, Theobroma cacao (the chocolate tree). Whole genome re-sequencing and transcriptome sequencing of a core collection of highly diverse cacao genotypes will provide the genetic information necessary to drive the discovery of genes critical for pathogen resistance. Functional analysis of these genes will test their role in resistance and set the stage for future translation of these basic findings to guiding more efficient breeding programs utilizing a wider array of genetic diversity. Importantly, the methods, tools, and knowledge gained will be directly applicable to discovery of genes underlying important traits in other crops, especially heterozygous perennials such as trees and many grasses which are not particularly amenable to approaches developed for the major annual crops such as corn and soybean. The basic evolutionary and functional principles that will be discovered can be generalized to most if not all crop plants. To promote and build genomics research capacity in developing countries and to promote inter-disciplinary cross-training the project will support scientific exchanges between project members and foreign collaborators, through postdoctoral, graduate and undergraduate student training at multiple institutions and will involve students from minority serving institutions. The results of this study will be made publicly available through electronic resources and publications.