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Apomixis and ploidy barrier suppress pollen-mediated gene flow in field grown transgenic turf and forage grass (Paspalum notatum Flüggé)

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Abstract

Bahiagrass (Paspalum notatum Flüggé) is the predominant forage grass in the southeastern US. The commercially important bahiagrass cultivar ‘Argentine’ is preferred for genetic transformation over sexual diploid cytotypes, since it produces uniform seed progeny through apomixis. Pseudogamous apomictic seed production in Argentine bahiagrass may contribute to transgene confinement. It is characterized by embryo development which is independent of fertilization of the egg cell, but requires fertilization with compatible pollen to produce the endosperm. Pollen-mediated gene transfer from transgenic, glufosinate-resistant apomictic bahiagrass as pollen donor at close proximity (0.5–3.5 m) with non-transgenic sexual or apomictic bahiagrass cultivars as pollen receptors was evaluated under field conditions. Hybridization frequency was evaluated by glufosinate herbicide resistance in >23,300 seedlings derived from open-pollinated (OP) pollen receptor plants. Average gene transfer between transgenic apomictic, tetraploid and sexual diploid bahiagrass was 0.03%. Herbicide-resistant hybrids confirmed by immuno-chromatographic detection of the PAT protein displayed a single copy bar gene identical to the pollen parent. Hybrids resulting from diploid pollen receptors were confirmed as triploids or aneu-triploids with significantly reduced vigor and seed set as compared to the parents. Transmission of transgenes to sexual bahiagrass is severely restricted by the ploidy difference between tetraploid apomicts and diploid sexual bahiagrass. Average gene transfer between transgenic apomictic tetraploid and non-transgenic, apomictic tetraploid bahiagrass was 0.17%, confirming a very low frequency of amphimixis in apomictic bahiagrass cultivars. While not providing complete transgene containment, gene transfer between transgenic apomictic and non-transgenic bahiagrass occurs at a much lower frequency than reported for other cross-pollinating or facultative apomictic grasses.

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Acknowledgments

We are grateful to the USDA-CSREES Biotechnology Risk Assessment Program for financial support. We appreciate Neal Benson (Genetics Institute, University of Florida) and Richard Fethiere (Agronomy Department, University of Florida) for their assistance with flow cytometry, Jeff Seib (Agronomy Department, University of Florida) for training Sukhpreet Sandhu in safe handling of radioisotopes, Carlos Alvarado (Agronomy Department, University of Florida) for excellent technical assistance and Hangning Zhang (Agronomy Department, University of Florida) for his help with molecular analysis. We like to thank Conrad Fafard Inc. Apopka, FL for the donation of Fafard potting mix.

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  1. Sukhpreet Sandhu

    Present address: Center for Applied Genetic Technologies, Institute of Plant Breeding Genetics and Genomics, University of Georgia, Athens, GA, USA

Authors and Affiliations

  1. Plant Molecular and Cellular Biology Program, Agronomy Department, Genetics Institute, University of Florida, IFAS, Gainesville, FL, USA

    Sukhpreet Sandhu, Kenneth H. Quesenberry & Fredy Altpeter

  2. North Florida Research and Education Center, University of Florida, IFAS, Marianna, FL, USA

    Ann R. Blount

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  1. Sukhpreet Sandhu
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Correspondence to Fredy Altpeter.

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Communicated by P. Heslop-Harrison.

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Sandhu, S., Blount, A.R., Quesenberry, K.H. et al. Apomixis and ploidy barrier suppress pollen-mediated gene flow in field grown transgenic turf and forage grass (Paspalum notatum Flüggé). Theor Appl Genet 121, 919–929 (2010). https://doi.org/10.1007/s00122-010-1360-3

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