2005 Progress Report: Molecular Improvement of an Environmentally Friendly Turfgrass

EPA Grant Number: R829479C017
Subproject: this is subproject number 017 , established and managed by the Center Director under grant R829479
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program
Center Director: Schumacher, Dorin
Title: Molecular Improvement of an Environmentally Friendly Turfgrass
Investigators: Altpeter, Fredy
Institution: University of Florida
EPA Project Officer: Lasat, Mitch
Project Period: January 1, 2004 through December 31, 2004 (Extended to December 31, 2007)
Project Period Covered by this Report: January 1, 2005 through December 31, 2006
RFA: The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program (2001) RFA Text |  Recipients Lists
Research Category: Targeted Research , Hazardous Waste/Remediation

Objective:

The objectives of this research are to: (1) develop a genetic transformation protocol for turf-type bahiagrass; and (2) modulate the bioactive gibberellic acid content in transgenic turf-type bahiagrass for improved turf characteristics and reduced mowing requirements.

Progress Summary:

Bahiagrass is extensively used in the southern United States for utility turf along highways and for residential lawns. The popularity of bahiagrass is attributed to its excellent persistence supported by drought tolerance, heat tolerance, insect- and disease-resistance, and nematode suppression. Genetic engineering of turf-type bahiagrass is desirable to improve turf quality and reduce the mowing frequency. The cultivar, Argentine, was selected for genetic transformation because it is a commercially important turf-type bahiagrass cultivar and its tetraploid, apomictic nature should result in uniformity of the transgenic progeny and transgene containment because of sexual incompatibility with diploid wildtype bahiagrasses. We developed an efficient protocol for tissue culture and transformation of Argentine bahiagrass using nptII as a selectable marker.

Using this protocol, several constructs containing different GA 2-oxidases (GA 2-oxidase1 and GA 2-oxidase8 from Arabidopsis) under control of two different, constitutive promoters have been constructed and introduced into bahiagrass. Table 1 presents a summary of the constructs created and the lines generated with each of them.

Table 1. Summary of the Constructs Created and Lines Generated

These expression cassettes were co-bombarded with a constitutive nptII expression cassette. So far, 54 independent lines have been regenerated from tissue cultures, following selection on paromomycin containing media and were established in soil. The transgenic nature of 40 lines has been confirmed by nptII Enzyme-Linked Immunosorbent Assay (ELISA). Transgenic lines are currently being assessed for the presence of the transgene by polymerase chain reaction (PCR) of genomic deoxyribonucleic acid (DNA) and the expression of GA 2-oxidase using real-time polymerase chain reaction (RT-PCR). So far, 17 lines were confirmed for GA 2-oxidase integration and 4 lines for GA 2 oxidase expression.

Transgenic lines have been established in the greenhouse and are being assessed for changes in plant height, tillering, and flowering as compared to the wildtype. A hydroponics set-up also is being used to compare root growth and shoot growth of the transgenic lines with wild-type plants. Transgenic lines with different degrees of dwarfing have been identified.

Future Activities:

Analysis of transgene expression by RT-PCR is in progress and will be completed in 2005. Selected transgenic lines with a dwarf phenotype will be analyzed by RT-PCR and Southern bloting for quantification of transgene expression and analysis of transgene copy number. The lines showing the most desirable phenotype will be evaluated in the field for persistence under mowing and reduced inputs.

Journal Articles:

No journal articles submitted with this report: View all 5 publications for this subproject

Supplemental Keywords:

sustainable industry, waste, agricultural engineering, bioremediation, environmental engineering, new technology, innovative technology, bioaccumulation, biodegradation, bioenergy, bioengineering, biotechnology, phytoremediation, plant biotechnology, molecular growth regulation, transgenic turfgrass, bahiagrass, dwarf, turf quality, mowing,, Scientific Discipline, TREATMENT/CONTROL, Sustainable Industry/Business, Genetics, Technology, New/Innovative technologies, Ecology and Ecosystems, Agricultural Engineering, bioengineering, transgenic plants, plant genes, bahiagrass, biotechnology, plant biotechnology, cloning, environmentally friendly turfgrass

Relevant Websites:

http://www.cpbr.org Exit

Progress and Final Reports:

Original Abstract
  • 2004 Progress Report
  • 2006
  • Final Report

  • Main Center Abstract and Reports:

    R829479    The Consortium for Plant Biotechnology Research, Inc., Environmental Research and Technology Transfer Program

    Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
    R829479C001 Plant Genes and Agrobacterium T-DNA Integration
    R829479C002 Designing Promoters for Precision Targeting of Gene Expression
    R829479C003 aka R829479C011 Biological Effects of Epoxy Fatty Acids
    R829479C004 Negative Sense Viral Vectors for Improved Expression of Foreign Genes in Insects and Plants
    R829479C005 Development of Novel Plastics From Agricultural Oils
    R829479C006 Conversion of Paper Sludge to Ethanol
    R829479C007 Enhanced Production of Biodegradable Plastics in Plants
    R829479C008 Engineering Design of Stable Immobilized Enzymes for the Hydrolysis and Transesterification of Triglycerides
    R829479C009 Discovery and Evaluation of SNP Variation in Resistance-Gene Analogs and Other Candidate Genes in Cotton
    R829479C010 Woody Biomass Crops for Bioremediating Hydrocarbons and Metals
    R829479C011 Biological Effects of Epoxy Fatty Acids
    R829479C012 High Strength Degradable Plastics From Starch and Poly(lactic acid)
    R829479C013 Development of Herbicide-Tolerant Energy and Biomass Crops
    R829479C014 Identification of Receptors of Bacillus Thuringiensis Toxins in Midguts of the European Corn Borer
    R829479C015 Coordinated Expression of Multiple Anti-Pest Proteins
    R829479C016 A Novel Fermentation Process for Butyric Acid and Butanol Production from Plant Biomass
    R829479C017 Molecular Improvement of an Environmentally Friendly Turfgrass
    R829479C018 Woody Biomass Crops for Bioremediating Hydrocarbons and Metals. II.
    R829479C019 Transgenic Plants for Bioremediation of Atrazine and Related Herbicides
    R829479C020 Root Exudate Biostimulation for Polyaromatic Hydrocarbon Phytoremediation
    R829479C021 Phytoremediation of Heavy Metal Contamination by Metallohistins, a New Class of Plant Metal-Binding Proteins
    R829479C022 Development of Herbicide-Tolerant Energy and Biomass Crops
    R829479C023 A Novel Fermentation Process for Butyric Acid and Butanol Production from Plant Biomass
    R829479C024 Development of Vectors for the Stoichiometric Accumulation of Multiple Proteins in Transgenic Crops
    R829479C025 Chemical Induction of Disease Resistance in Trees
    R829479C026 Development of Herbicide-Tolerant Hardwoods
    R829479C027 Environmentally Superior Soybean Genome Development
    R829479C028 Development of Efficient Methods for the Genetic Transformation of Willow and Cottonwood for Increased Remediation of Pollutants
    R829479C029 Development of Tightly Regulated Ecdysone Receptor-Based Gene Switches for Use in Agriculture
    R829479C030 Engineered Plant Virus Proteins for Biotechnology