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RESEARCH ARTICLE

Assessment of gene flow in white clover (Trifolium repens L.) under field conditions in Australia using phenotypic and genetic markers

J. A. De Lucas A C D , J. W. Forster A C D , K. F. Smith B C D and G. C. Spangenberg A C D E
+ Author Affiliations
- Author Affiliations

A Department of Primary Industries, Biosciences Research Division, Victorian Agribiosciences Centre, La Trobe University R&D Park, Bundoora, Vic. 3083, Australia.

B Department of Primary Industries, Biosciences Research Division, Hamilton Centre, Mount Napier Road, Hamilton, Vic. 3300, Australia.

C Molecular Plant Breeding Cooperative Research Centre, Australia, 1 Park Drive, Bundoora, Vic. 3083, Australia.

D La Trobe University, Bundoora, Vic. 3086, Australia.

E Corresponding author. Email: german.spangenberg@dpi.vic.gov.au

Crop and Pasture Science 63(2) 155-163 https://doi.org/10.1071/CP11224
Submitted: 22 August 2011  Accepted: 2 February 2012   Published: 26 March 2012

Abstract

White clover is one of the most important pasture legumes in global temperate regions. It is an outcrossing, insect-pollinated species with gene flow occurring naturally between plants. A 2-year study was conducted to assess the relationship between gene flow and physical distance in white clover under field conditions in southern Australia. White clover plants exhibiting a red leaf mark phenotypic trait acted as pollen donors to recipient plants lacking leaf markings at distances up to 200 m distant from the donor plants. Progeny were scored for the dominant red-leafed phenotype and gene flow was modelled. Paternity was confirmed using simple sequence repeat markers. A leptokurtic pattern of gene flow was observed under conditions designed to measure maximised gene flow with the majority of pollination occurring in the first 50 m from the donor pollen source. The combined use of simple sequence repeat and visual markers confirmed that there was also a white clover pollen source in addition to the donor plants. This research confirms the difficulty in ensuring absolute containment of gene flow in an outcrossing species grown in an environment when endemic populations are known to exist.

Additional keywords: field-design, pasture legume, phenotype, red leaf mark, single sequence repeat, Trifolium repens.


References

Abberton MT (2007) Interspecific hybridization in the genus Trifolium. Plant Breeding 126, 337–342.
Interspecific hybridization in the genus Trifolium.Crossref | GoogleScholarGoogle Scholar |

Atwood SS (1940) Genetics of cross-incompatibility among self-incompatible plants of Trifolium repens. Journal - American Society of Agronomy 32, 955–968.
Genetics of cross-incompatibility among self-incompatible plants of Trifolium repens.Crossref | GoogleScholarGoogle Scholar |

Atwood SS (1942) Oppositional alleles causing cross-incompatibility in Trifolium repens. Genetics 27, 333–338.

Becker HC, Damgaard C, Karlsson B (1992) Environmental variation for outcrossing rate in rapeseed (Brassica napus). Theoretical and Applied Genetics 84, 303–306.
Environmental variation for outcrossing rate in rapeseed (Brassica napus).Crossref | GoogleScholarGoogle Scholar |

Brewbaker JL (1954) Incompatibility in autotetraploid Trifolium repens L. I. Competition and self-compatibility. Genetics 39, 307–316.

Brewbaker JL (1955) V-leaf markings in white clover. The Journal of Heredity 46, 115–123.

Clifford PTP, Sparks GA, Woodfield DR (1996) The intensifying requirements for cultivar change. Grasslands Research and Practice Series 6, 19–24.

Corkill L (1971) Leaf markings in white clover. The Journal of Heredity 62, 307–310.

Cunliffe KV, Vecchies AC, Jones ES, Kearney GA, Forster JW, Spangenberg GC, Smith KF (2004) Assessment of gene flow using tetraploid genotypes of perennial ryegrass (Lolium perenne L.). Australian Journal of Agricultural Research 55, 389–396.
Assessment of gene flow using tetraploid genotypes of perennial ryegrass (Lolium perenne L.).Crossref | GoogleScholarGoogle Scholar |

Damgaard C, Kjellsson G (2005) Gene flow of oilseed rape (Brassica napus) according to isolation distance and buffer zone. Agriculture, Ecosystems & Environment 108, 291–301.
Gene flow of oilseed rape (Brassica napus) according to isolation distance and buffer zone.Crossref | GoogleScholarGoogle Scholar |

Damgaard C, Simonsen V, Osborne J (2008) Prediction of pollen-mediated gene flow between fields of red clover (Trifolium pratense). Environmental Modeling and Assessment 13, 483–490.
Prediction of pollen-mediated gene flow between fields of red clover (Trifolium pratense).Crossref | GoogleScholarGoogle Scholar |

De Marchis F, Bellucci M, Arcioni S (2003) Measuring gene flow from two birdsfoot trefoil (Lotus corniculatus) field trials using transgenes as tracer markers. Molecular Ecology 12, 1681–1685.
Measuring gene flow from two birdsfoot trefoil (Lotus corniculatus) field trials using transgenes as tracer markers.Crossref | GoogleScholarGoogle Scholar |

Eastham K, Sweet J (2002) Genetically modified organisms (GMOs): the significance of gene flow through pollen transfer. European Environment Agency, Environmental Issue Report No. 28, Copenhagen.

Emmerling M, Chu P, Smith K, Kalla R, Spangenberg G (2004) Field evaluation of transgenic white clover with AMV immunity and development of elite transgenic germplasm. In ‘Molecular breeding of forage and turf’. (Eds A Hopkins, Y Wang, R Mian, M Sledge, E Barker) pp. 359–366. (Kluwer Academic Press: Dordrecht, the Netherlands)

Godfree RC, Vivian LM, Lepschi BJ (2006) Risk assessment of transgenic virus-resistant white clover: non-target plant community characterisation and implications for field trial design. Biological Invasions 8, 1159–1178.
Risk assessment of transgenic virus-resistant white clover: non-target plant community characterisation and implications for field trial design.Crossref | GoogleScholarGoogle Scholar |

Goodman RD, Williams AE (1994) Honeybee pollination of white clover (Trifolium repens L.) cv. Haifa. Australian Journal of Experimental Agriculture 34, 1121–1123.
Honeybee pollination of white clover (Trifolium repens L.) cv. Haifa.Crossref | GoogleScholarGoogle Scholar |

Hokanson SC, Grumet R, Hancock JF (1997) Effect of border rows and trap/donor ratios on pollen-mediated gene movement. Ecological Applications 7, 1075–1081.
Effect of border rows and trap/donor ratios on pollen-mediated gene movement.Crossref | GoogleScholarGoogle Scholar |

Hovin AW, Gibson PB (1961) A red leaf marking in white clover. The Journal of Heredity 52, 295–296.

Hüsken A, Dietz-Pfeilstetter A (2007) Pollen-mediated intraspecific gene flow from herbicide resistant oilseed rape (Brassica napus L). Transgenic Research 16, 557–569.
Pollen-mediated intraspecific gene flow from herbicide resistant oilseed rape (Brassica napus L).Crossref | GoogleScholarGoogle Scholar |

Johnson PG, Larson SR, Anderton AL, Patterson JT, Cattani DJ, Nelson EK (2006) Pollen-mediated gene flow from Kentucky bluegrass under cultivated field conditions. Crop Science 46, 1990–1997.
Pollen-mediated gene flow from Kentucky bluegrass under cultivated field conditions.Crossref | GoogleScholarGoogle Scholar |

Lavigne C, Godelle B, Reboud X, Gouyon PH (1996) A method to determine the mean pollen dispersal of individual plants growing within a large pollen source. Theoretical and Applied Genetics 93, 1319–1326.
A method to determine the mean pollen dispersal of individual plants growing within a large pollen source.Crossref | GoogleScholarGoogle Scholar |

Lavigne C, Klein EK, Vallée P, Pierre J, Godelle B, Renard M (1998) A pollen-dispersal experiment with transgenic oilseed rape. Estimation of the average pollen dispersal of an individual plant within a field. Theoretical and Applied Genetics 96, 886–896.
A pollen-dispersal experiment with transgenic oilseed rape. Estimation of the average pollen dispersal of an individual plant within a field.Crossref | GoogleScholarGoogle Scholar |

Manasse RS (1992) Ecological risks of transgenic plants: effects of spatial dispersion on gene flow. Ecological Applications 2, 431–438.
Ecological risks of transgenic plants: effects of spatial dispersion on gene flow.Crossref | GoogleScholarGoogle Scholar |

Marshall AH, Michaelson-Yeates TPT, Williams IH (1999) ‘How busy are bees – modelling the pollination of clover.’ IGER Innovations 3. (Aberystwyth University: Aberystwyth, UK)

Messeguer J, Fogher C, Guiderdoni E, Marfà V, Català MM, Baldi G, Melé E (2001) Field assessments of gene flow from transgenic to cultivated rice (Oryza sativa L.) using a herbicide resistance gene as tracer marker. Theoretical and Applied Genetics 103, 1151–1159.
Field assessments of gene flow from transgenic to cultivated rice (Oryza sativa L.) using a herbicide resistance gene as tracer marker.Crossref | GoogleScholarGoogle Scholar |

Morris WF (1993) Predicting the consequence of plant spacing and biased movement for pollen dispersal by honey bees. Ecology 74, 493–500.
Predicting the consequence of plant spacing and biased movement for pollen dispersal by honey bees.Crossref | GoogleScholarGoogle Scholar |

Morris WF, Kareiva PM, Raymer PL (1994) Do barren zones and pollen traps reduce gene escape from transgenic crops? Ecological Applications 4, 157–165.
Do barren zones and pollen traps reduce gene escape from transgenic crops?Crossref | GoogleScholarGoogle Scholar |

Pierre J (2001) The role of honeybees (Apis mellifera) and other insect pollinators in gene flow between oilseed rape (Brassica napus) and wild radish (Raphanus raphanistrum). Acta Horticulturae 561, 47–51.

Ramsay G, Thompson CE, Nielson S, Mackay GR (1999) Honeybees as vectors of GM oilseed rape pollen. In ‘Gene flow and agriculture: relevance for transgenic crops’. (Ed. PJW Lutman) pp. 209–224. (British Crop Protection Council: Staffordshire, UK)

Ramsay G, Thompson C, Squire G (2003) Quantifying landscape-scale gene flow in oilseed rape. Department for Environment, Food and Rural Affairs Project No. RG0216, London.

Rieger MA, Lamond M, Preston C, Powles SB, Roush RT (2002) Pollen-mediated movement of herbicide resistance between commercial canola fields. Science 296, 2386–2388.
Pollen-mediated movement of herbicide resistance between commercial canola fields.Crossref | GoogleScholarGoogle Scholar |

Rognli OA, Nilsson NO, Nurminiemi M (2000) Effects of distance and pollen competition on gene flow in the wind-pollinated grass Festuca pratensis Huds. Heredity 85, 550–560.
Effects of distance and pollen competition on gene flow in the wind-pollinated grass Festuca pratensis Huds.Crossref | GoogleScholarGoogle Scholar |

Saeglitz C, Pohl M, Bartsch D (2000) Monitoring gene flow from transgenic sugar beet using cytoplasmic male-sterile bait plants. Molecular Ecology 9, 2035–2040.
Monitoring gene flow from transgenic sugar beet using cytoplasmic male-sterile bait plants.Crossref | GoogleScholarGoogle Scholar |

Scheffler JA, Parkinson R, Dale PJ (1993) Frequency and distance of pollen dispersal from transgenic oilseed rape (Brassica napus). Transgenic Research 2, 356–364.
Frequency and distance of pollen dispersal from transgenic oilseed rape (Brassica napus).Crossref | GoogleScholarGoogle Scholar |

St Amand PC, Skinner DZ, Peaden RN (2000) Risk of alfalfa transgene dissemination and scale-dependent effects. Theoretical and Applied Genetics 101, 107–114.
Risk of alfalfa transgene dissemination and scale-dependent effects.Crossref | GoogleScholarGoogle Scholar |

Staniland BK, McVetty PBE, Friesen LF, Yarrow S, Thiel P, Freyssinet G, Freyssinet M (2000) Effectiveness of border areas in confining the spread of transgenic Brassica napus pollen. Canadian Journal of Plant Science 80, 521–526.
Effectiveness of border areas in confining the spread of transgenic Brassica napus pollen.Crossref | GoogleScholarGoogle Scholar |

Van Deynze AE, Fitzpatrick S, Hammon B, McCaslin M, Putnam D, Hammon B, Teuber L, Undersander D (2008) ‘Gene flow in alfalfa: biology, mitigation, and potential impact on production.’ (Council for Agricultural Science and Technology (CAST): Ames, IA)

Velterop O (2000) Effects of fragmentation on pollen and gene flow in insect-pollinated plant populations. Doctorate Thesis, University of Groningen, The Netherlands.

Wang ZY, Lawrence R, Hopkins A, Bell J, Scott M (2004) Pollen-mediated transgene flow in the wind-pollinated grass species tall fescue (Festuca arundinacea Schreb.). Molecular Breeding 14, 47–60.
Pollen-mediated transgene flow in the wind-pollinated grass species tall fescue (Festuca arundinacea Schreb.).Crossref | GoogleScholarGoogle Scholar |

Watrud LS, Lee EH, Fairbrother A, Burdick C, Reichman JR, Bollman M, Storm M, King G, Van de Water PK (2004) Evidence for landscape-level, pollen-mediated gene flow from genetically modified creeping bentgrass with CP4 EPSPS as a marker. Proceedings of the National Academy of Sciences of the United States of America 101, 14 533–14 538.
Evidence for landscape-level, pollen-mediated gene flow from genetically modified creeping bentgrass with CP4 EPSPS as a marker.Crossref | GoogleScholarGoogle Scholar |

Weaver N (1965) Foraging behavior of honeybees on white clover. Insectes Sociaux 12, 231–240.
Foraging behavior of honeybees on white clover.Crossref | GoogleScholarGoogle Scholar |

Woodfield DR, Clifford PTP, Baird IJ, Cousins GR (1995) Gene flow and estimated isolation requirements for transgenic white clover. In ‘Proceedings of the 3rd International Symposium on the Biosafety Results of Field Tests of Genetically Modified Plants and Microorganisms’. University of California, Oakland, USA. (Ed. DD Jones) pp. 509–514.