Final 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)
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 are:

  1. Development of a genetic transformation protocol for turf-type bahiagrass.
  2. Modulation of the bioactive gibberellic acid content in transgenic turf-type bahiagrass for improved turf characteristics and reduced mowing requirements.

Summary/Accomplishments (Outputs/Outcomes):

Objective # 1: Development of a genetic transformation protocol for turf-type bahiagrass.

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 due to sexual incompatibility with diploid wildtype bahiagrasses. We developed an efficient tissue culture system for regeneration of turf-type bahiagrass cv. Argentine from mature seed -derived callus. Mature seed explants are preferred over other explants because they are readily available year-round, and the explant preparation prior to culture initiation is less labor intensive.

An efficient biolistic gene transfer protocol for turf-type bahiagrass (cv. Argentine) was developed. Transfer of a constitutive nptII, bar, or EPSPS expression cassette into seed- derived callus cultures was followed by selection with paromomycin sulfate, bialaphos, or glyphosate respectively during callus subculture and / or regeneration. Transgenic plants were confirmed expressing bar, nptII, or EPSPS. Consequently, all three selectable markers supported the generation of transgenic plants. The comparison of three different selectable marker genes revealed significant differences in transformation efficiency. Immuno detection of transgene expression revealed that one to five transgenic plants were generated from the three different selectable marker genes per 10 bombarded Petri dishes with 20 callus pieces each. Transgenic plants were transferred to soil in approximately 17 to 20 weeks from the initiation of mature seed cultures. Transgenic nature of regenerated plants was confirmed with PCR of genomic DNA, using primers annealing to the transgene expression cassette and by immuno assays (ELISA or Quicksticks) of leaf protein extracts. So far, more than 100 trangenic plants have been generated using the superior nptII selectable marker. Stability of transgene expression in sexual progeny plants was confirmed with ELISA.

Objective # 2: Modulation of the bioactive gibberellic acid content in transgenic turf-type bahiagrass for improved turf characteristics and reduced mowing requirements.

The established transformation protocol was used to introduce a gibberellic acid catabolizing enzyme (GA-2-oxidase) under control of different regulatory promoter elements to achieve a reduction of bioactive gibberellic acid, which is expected to result in dwarfing, reduced mowing frequency, darker green color, and enhanced drought and shade tolerance. The following vectors have been completed and were introduced into bahiagrass: 35S promoter - hsp70intron - GA2ox - NOS; Ubiquitin promoter - ubi1intron - GA2ox - NOS. So far, more than 20 transgenic plants showing detectable expression (RT-PCR) of the GA-2-oxidase and the selectable nptII gene (ELISA) have been regenerated. The plants show an altered phenotype with growth reduction ranging from 10 to 90 percent, reduced internode length, darker green color, and more tillering under controlled environment conditions. In the near future, we will analyze turf characteristics of the transgenic plants under field conditions.

Field evaluation of the transgenic turfgrass will allow us to correlate transgene expression with turf quality and to identify superior transgenic lines. The goal is to identify the lines that combine the most desirable phenotype with persistence under mowing and good seed production.


Journal Articles on this Report : 2 Displayed | Download in RIS Format

Other subproject views: All 5 publications 2 publications in selected types All 2 journal articles
Other center views: All 211 publications 48 publications in selected types All 44 journal articles
Type Citation Sub Project Document Sources
Journal Article Altpeter F, Positano M. Efficient plant regeneration from mature seed derived embryogenic callus of turf-type bahiagrass (Paspalum notatum Flugge). International Turfgrass Society Research Journal 2005;10(Part 1):479-484. R829479C017 (Final)
not available
Journal Article Altpeter F, James VA. Genetic transformation of turf-type bahiagrass (Paspalum notatum Flugge) by biolistic gene transfer. International Turfgrass Society Research Journal 2005;10(Part 1):485-489. R829479C017 (Final)
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  • Supplemental Keywords:

    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://cris.csrees.usda.gov/ exit EPA

    Progress and Final Reports:

    Original Abstract
  • 2004 Progress Report
  • 2005 Progress Report
  • 2006

  • 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