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Risk assessment of transgenic apomictic tetraploid bahiagrass, cytogenetics, breeding behavior and performance of intra-specific hybrids

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Abstract

Pollen-mediated gene transfer from stress tolerant or herbicide-resistant transgenic plants may cause environmental or agronomic problems. Apomictic seed production found in some bahiagrass cultivars may serve as a natural transgene containment system. Under greenhouse conditions, the average gene transfer frequency from an herbicide-resistant apomictic tetraploid to a population of sexual diploid bahiagrass genotypes or apomictic tetraploid bahiagrass was 0.16% when the transgenic pollen donor was placed at 0.5–1.5 m distance from the non-transgenic pollen receptors. The herbicide-resistant hybrids were characterized for transgene integration, expression and ploidy, by Southern blot analysis, immuno-chromatography and flow cytometry, respectively. Hybrids resulting from open pollination of non-transgenic diploid female plants with transgenic tetraploid male plants were triploids or near-triploids, with 2n = 26–34. These hybrids displayed a wide range of phenotypic variability, including some non-persistent or non-flowering dwarf-type hybrids with good vigor, or hybrids with vegetative growth similar to non-transgenic plants, but with significantly reduced seed set. Non-flowering aneu-triploids with good vigor/field performance will provide the highest level of transgene containment. Embryo sac analysis of pollinated spikelets confirmed a high proportion of aborted ovules. An apospory-linked RFLP marker was detected in 13 of the 15 near-triploid hybrids. All flowering aneuploid hybrids displayed significantly reduced seed set, and none of the sexual near-triploid hybrids produced any seeds. All tetraploid gene transfer events carried the apospory-linked RFLP marker, suggesting that despite the presence of the aposporus locus, a low degree of sexuality co-exists in apomictic tetraploid cultivars. Thus, tetraploid apomictic bahiagrass does not provide complete transgene containment, although intra-specific gene transfer is drastically reduced compared to sexually reproducing perennial grasses.

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Notes

  1. Clonal plants for the second and third experiment set were generated by clonal propagation of a single seedling of cultivar Pensacola in liquid culture media containing 4.3 g l−1 MS salts with vitamins (Murashige and Skoog 1962), 30 g l−1 sucrose and 0.3 mg l−1 6-benzylaminopurine (BAP).

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Acknowledgments

This project was funded by USDA-CSREES Biotechnology Risk Assessment program. We are grateful to Carlos A. Acuna (Agronomy Department, University of Florida) for his help with embryo sac analysis. We are thankful to Neal Benson (Genetics Institute, University of Florida) and Richard Fethiere (Agronomy Department, University of Florida) for their assistance with flow cytometric analysis, 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.

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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

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

    Sukhpreet Sandhu, Victoria A. James, Kenneth H. Quesenberry & Fredy Altpeter

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  1. Sukhpreet Sandhu
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  2. Victoria A. James
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Correspondence to Fredy Altpeter.

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Sandhu, S., James, V.A., Quesenberry, K.H. et al. Risk assessment of transgenic apomictic tetraploid bahiagrass, cytogenetics, breeding behavior and performance of intra-specific hybrids. Theor Appl Genet 119, 1383–1395 (2009). https://doi.org/10.1007/s00122-009-1142-y

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