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Chantilly, VA, United States

Muthukumar B.,University of Tennessee at Knoxville | Joyce B.L.,University of Tennessee at Knoxville | Elless M.P.,Edenspacec Inc | Stewart Jr. C.N.,University of Tennessee at Knoxville
Plant Physiology | Year: 2013

Ferns (Pteridophyta) are very important members of the plant kingdom that lag behind other taxa with regards to our understanding of their genetics, genomics, and molecular biology. We report here, to our knowledge, the first instance of stable transformation of fern with recovery of transgenic sporophytes. Spores of the arsenic hyperaccumulating fern Pteris vittata and tetraploid 'C-fern Express' (Ceratopteris thalictroides) were stably transformed by Agrobacterium tumefaciens with constructs containing the P. vittata actin promoter driving a GUSPlus reporter gene. Reporter gene expression assays were performed on multiple tissues and growth stages of gametophytes and sporophytes. Southern-blot analysis confirmed stable transgene integration in recovered sporophytes and also confirmed that no plasmid from A. tumefaciens was present in the sporophyte tissues. We recovered seven independent transformants of P. vittata and four independent C. thalictroides transgenics. Inheritance analyses using b-glucuronidase (GUS) histochemical staining revealed that the GUS transgene was stably expressed in second generation C. thalictroides sporophytic tissues. In an independent experiment, the gusA gene that was driven by the 2× Cauliflower mosaic virus 35S promoter was bombarded into P. vittata spores using biolistics, in which putatively stable transgenic gametophytes were recovered. Transformation procedures required no tissue culture or selectable marker genes. However, we did attempt to use hygromycin selection, which was ineffective for recovering transgenic ferns. This simple stable transformation method should help facilitate functional genomics studies in ferns. © 2013 American Society of Plant Biologists. All Rights Reserved. Source


Grant
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 150.00K | Year: 2015

DESCRIPTION provided by applicant Agricultural application of arsenate based pesticides from the s through s left significant acreage of arsenic residues particularly on land used for apple potato and blueberry farming Arsenate was applied at rates up to kg ha in most fruit orchards until insecticides such as DDT were introduced the late s resulting in contamination over large areas These wide area and smaller localized areas of arsenic contaminated soils are difficult and expensive to remediate via conventional means An alternative approach is phytoremediation using living plants to extract and concentrate the element from contaminated soils and waters The arsenic accumulating Edenfern tm plants have been used commercially to decrease arsenic concentrations at a number of sites Because the ferns are native to semi tropical environments their use in northern or temperate climates is restricted to annual plantings that increase cost Endophytic bacteria and fungi that colonize specific plants have been shown to confer tolerance to adverse conditions improve plant nutrient utilization increase disease resistance and facilitate degradation of soil and water contaminants such as TCE and PAHs An understanding of the many benefits conferred by endophytic organisms is still developing and recently the Doty laboratory isolated endophytes from plants growing on arsenic contaminated soils within the Tacoma Smelter Plume in Washington State These bacterial endophytes have shown an unusual tolerance to arsenic and may provide improved arsenic accumulation in phytoremediation applications This project seeks to address wide area arsenic contamination through the use of conventional non transgenic endophytes that improve arsenic tolerance and uptake in woody biomass crops such as willow for phytoremediation The use of novel endophytes isolated from native plants found on arsenic contaminated soils generates a technology approach that will allow a variety of crops and cropping systems to be used for phytoremediation The Phase I approach provides a strong basis for Phase II work which if successful will provide site managers with an invaluable low cost tool for removal of arsenic from contaminated soils PUBLIC HEALTH RELEVANCE This Phase I project seeks to reduce the cost of remediating soils contaminated with arsenic by developing a plant endophyte system using willow plants with an improved ability to tolerate and accumulate arsenic from the soil The project focuses on using naturally occurring bacterial endophytes found in plants growing on arsenic contaminated sites to increase tolerance to arsenic in a crop plant suited for phytoremediation to allow cost effective treatment of wide area contamination


Grant
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 150.00K | Year: 2011

DESCRIPTION (provided by applicant): Trichloroethylene (TCE), one of the most common groundwater pollutants, is a known hepatotoxin and carcinogen. It has been widely used by industry and the military as a degreaser for metal parts: according to the Agencyfor Toxic Substances and Disease Registry, more than eight hundred Superfund sites in the United States are contaminated with TCE. Poplar, which can take up and degrade TCE, is an attractive plant for phytoremediation of TCE and other organic contaminantsdue to its high growth rate, extensive root system, high rates of water uptake from the soil, and ease of genetic manipulation. While transgenic poplar for improved TCE degradation has been successfully field tested, there are significant regulatory and breeding hurdles preventing the large-scale use of this technology. Recently researchers have determined the potential of endophytes, symbiotic bacteria and fungi that live within plant cells, to break down organic contaminants and improve the phytoremediation capability of non-transgenic plants. Unlike other microbes that have been used for phytoremediation, endophytes live within the plant and therefore are expected to persist better at the site, continuing to degrade TCE as long as their plant partner survives. The laboratory of Dr. Sharon Doty is one of the pioneers in studying endophytes to improve plant growth and health, having worked in this field for over a decade. Recently her laboratory has isolated endophytes from poplar growing in sites contaminated with TCE and other organic pollutants. Some of these microbes exhibit high rates of TCE degradation when grown in the lab. Her laboratory is currently developing methods to inoculate these TCE-degrading endophytes into poplar and will work with Edenspace on this SBIR project to demonstrate that the new poplar/endophyte systems have significantly better TCE phytoremediation performance than control poplar. Edenspace will also develop molecular markers to identify the specific endophytes, in order to confirm in a field test that the microbes can persist in the poplar for months. Upon successful completion of this SBIR project Edenspace will partner with Geosyntec, a leading environmental engineering firm, to introduce this novel technology to the remediation industry. PUBLIC HEALTH RELEVANCE: Trichloroethylene (TCE) is a known carcinogen and significant environmental contaminant as a result of to extensive use by the military and industry. Poplar trees have been used as a natural method to remove TCE from contaminated groundwater. In this project scientists from Edenspace and the University of Washington will utilize recently identified microbes that can act with the poplar to greatly increase TCE removal rates. Development of this improved remediation system will reduce exposure of humans and wildlife to TCE.


Grant
Agency: Department of Agriculture | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 460.00K | Year: 2011

The U.S. population has a deficit of dietary calcium that represents a serious challenge to public health. Many consumers, however, find current methods of increasing dietary calcium to be inconvenient, costly, or unpalatable. This SBIR Phase II proposal seeks to increase calcium levels in lettuce, a specialty crop that already comprises a significant part of the average U.S. diet. Successful Phase I results included demonstration of the ability to provide up to 150 mg of calcium per serving of Bibb lettuce, a level high enough to qualify the lettuce as a "good" source of calcium under FDA guidelines and more than eight times the amount of calcium available in commercial lettuce today, using a combination of bioengineering and novel hydroponic techniques. Moreover, most of the calcium in the lettuce is present in a bioavailable form that is efficiently processed by the body. In Phase II, elite commercial lettuce varieties provided by a leading commercial seed company will be transformed, hydroponic growing protocols will be optimized, a pilot demonstration will be conducted at a commercial hydroponic lettuce grower, and product quality testing including taste testing will be conducted by a university partner. Successful completion of the project will lay the foundation for introduction of a new value-added crop that provides better nutrition for consumers and a new source of income for agricultural producers. Planned commercialization activities include marketing of the calcium-fortified lettuce at regional farmers' markets, followed by nationwide distribution through grocery chains and specialty food stores.


Patent
Edenspacec Inc | Date: 2010-04-30

The present invention is directed, among other things, to using secondary metabolites in the mevalonate pathway (such as, for example, HMG) and/or structurally related compounds to mediate biological activities (e.g., for therapeutic applications) and/or as diagnostic agents. In some embodiments, the biological activities comprise one or more pleiotropic effects of statins (such as, for example, angiogenesis, promoting vascular function, anti-inflammatory action, immunomodulation, etc.). Also provided are methods of screening for mevalonate pathway secondary metabolites, methods of producing HMG, and methods of diagnosing comprising measuring amount of mevalonate pathway secondary metabolites.

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