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GMO’s and GENE FLOW: A PLANT BREEDING PERSPECTIVE Kendall R. Lamkey Department of Agronomy Iowa State University Ames, IA 50010 http://www.agron.iastate.edu/corn 9-13-2002 Purdue 1 Outline • Conventional vs. Transgenic Crop Improvement • Gene Flow • Pollen Movement • Current APHIS Regulations • What Can Be Done • Conclusions 9-13-2002 Purdue 2 Conventional Plant Breeding • The art and science of plant improvement – Gene Exchange only occurs between sexually compatible species – Most of the genetic variation is created through recombination (crossing) – Selection is conducted by measuring plant characteristics (yield, grain moisture, etc) – Genes under selection are unknown – Little is known about the changes that occur at the DNA level with selection 9-13-2002 Purdue 3 Transgenics • The human mediated insertion of a synthetic gene or genes into a plant – Usually involves transfer of genes between sexually incompatible species – The gene produces a specific protein of interest – either as an input or output trait – Often referred to as “precise”, because we know the DNA sequence and gene product – Gene insertion into the genome is usually into a random location – Effect on other plant characteristics is unknown 9-13-2002 Purdue 4 Gene Discovery Gene Discovery: Where do the genes come from? • Basic research in plant physiology and biochemistry • Genome sequencing efforts such as the human genome project X ? 9-13-2002 Purdue 5 Transformation Parts of a transgene The promoter • Usually taken from a corn gene • Determines where and when the transgene will function • The gene can be Tissue Specific or Constitutive P(BLA) The coding sequence - r-zein pro Determines what the gene does APr Porcine lactalbumin 3'UTR1 pAHC25/zein pro/lactal Must be modified to resemble a corn gene 7320 bp ORI P(LAC) 3'UTR1 Ubi pro BAR 9-13-2002 Purdue 6 “Gene Gun” Backcrossing R +/+ BC3F1 (93.75%) ½ +/+ : ½ +/Bt R +/+ x D Bt/Bt F1 (50%) x +/Bt BC3F2 (93.75%) R BC1F1 (75%) +/+ x ½ +/+ : ½ +/Bt ¼ +/+ : ½ +/Bt : ¼ Bt/Bt BC3F3 Bt/Bt R BC2F1 (87.5%) +/+ x ½ +/+ : ½ +/Bt BC3F1 (93.75%) ½ +/+ : ½ +/Bt 9-13-2002 Purdue 9 Conventional vs. Transgenic • Genomes are fluid and dynamic – not stable and static • Difficult to distinguish between effects of conventional breeding and a transgene insertion event 9-13-2002 Purdue 10 Conventional vs. Transgenic • Transgenes result in novel phenotypes, but present no new categories of risks • Specific traits introduced by both conventional and transgenic approaches can pose unique risks • It is the product and not the process we should be concerned about NCR (2002) Environmental effects of transgenic plants 9-13-2002 Purdue 11 Gene flow • Gene flow is movement between groups that results in genetic exchange – Gene flow always results in contamination – Contamination does not result in gene flow, unless it is followed by gene exchange 9-13-2002 Purdue 12 Gene Flow • Seed Dispersal – Spillage – Seed shattering – Mechanical mixing • Horizontal Transfer – nonsexual transfer of genetic material from one organism to another • Pollen Movement 9-13-2002 Purdue 13 Pollen Movement • Crop to wild – Grain crops have no wild relatives in the US – Needs to be assessed on a crop by crop basis • Crop to weed – Must be assessed on a crop by crop basis • Crop to crop – Our primary concern 9-13-2002 Purdue 14 Pollen Movement • Primarily an issue with crosspollinated crops • Dependent on environmental conditions and the physical characteristics of the pollen 9-13-2002 Purdue 15 Pollen Movement - Corn • Characteristics of Corn – One plant produces 18,000,000 to 25,000,000 pollen grains – One plant theoretically has enough pollen to pollinate an acre of corn – There have been surprisingly few studies of pollen movement in corn, especially long distance movement 9-13-2002 Purdue 16 Pollen Movement - Corn M. Garcia C., J. Figueroa M., R. Gomez L., R. Townsend, and J. Schoper. 1998. Pollen control during transgenic hybrid maize development in Mexico. Crop Science 38:1597 9-13-2002 Purdue 17 Pollen Movement - Corn 19 Meters Yellow Corn Tassels Removed 30 Meters White Corn 9-13-2002 Plants With Yellow Seeds Purdue 18 Pollen Movement - Corn • Spatial isolation resulted in 0.01% contamination from a pollen source > 184 m from the field • Isolation in space and time resulted in no contamination • Pollen movement is best controlled when transgenics are used as females • Study is relevant only to breeding scale studies • Difficult to apply results from Mexico to US M. Garcia C., J. Figueroa M., R. Gomez L., R. Townsend, and J. Schoper. 1998. Pollen control during transgenic hybrid maize development in Mexico. Crop Science 38:1597 9-13-2002 Purdue 19 Pollen Movement - Corn • Four recommendations – Use transgenic plants as females and remove tassels prior to pollen shed – Spatial isolation > 185m – Use temporal isolation by planting 1 or 2 weeks later than surrounding corn – Scout and destroy sexually compatible plants in the area of presumed pollen movement M. Garcia C., J. Figueroa M., R. Gomez L., R. Townsend, and J. Schoper. 1998. Pollen control during transgenic hybrid maize development in Mexico. Crop Science 38:1597 9-13-2002 Purdue 20 Pollen Movement - Corn S. Luna V., J. Figueroa M., B. Baltazar M., R. Gomez L., R. Townsend, and J. B. Schoper. 2001. Maize Pollen Longevity and Distance Isolation Requirements for Effective Pollen Control. Crop Sci. 41:1551–1557 9-13-2002 Purdue 21 Pollen Movement - Corn S. Luna V., J. Figueroa M., B. Baltazar M., R. Gomez L., R. Townsend, and J. B. Schoper. 2001. Maize Pollen Longevity and Distance Isolation Requirements for Effective Pollen Control. Crop Sci. 41:1551–1557 9-13-2002 Purdue 22 Pollen Movement - Corn • It was clear maize pollen does not remain viable in the atmosphere for longer than 2 h near San Jose del Valle. • Actual measurements of pollen movement as measured by outcrossing from research scale plantings of material, indicated that viable pollen was barely detectable at 200 m and nonexistant at 300 m from the pollen source. 9-13-2002 Purdue 23 Pollen Movement - Corn • What is low? – Assume 1% contamination (100 0f 10,000 kernels – 1 lb corn = 1800 kernels – 1 bushel corn = 100,800 kernels – 1 semi = 750 bushels = 75,600,000 – 756,000 contaminating kernels per semi – Or 7.5 bushels / semi 9-13-2002 Purdue 24 What Can Be Done? • Barriers to Pollen Movement – Physical – Biological – Mechanical – Spatial – Temporal 9-13-2002 Purdue 25 What Can Be Done? • Physical barriers physically prevent pollen movement – Glass House – Wind Breaks – Mesh Nets 9-13-2002 Purdue 26 What Can Be Done? • Biological Barriers use genetic or molecular aspects of the plant itself – Apomixis – Male Sterility – Trangenes that render pollen grains in which they are expressed inviable – Terminator Technology 9-13-2002 Purdue 27 What Can Be Done? • Mechanical Barriers involves removal of the pollen producing structures – Removal of tassels – Perhaps use of gameticides 9-13-2002 Purdue 28 What Can Be Done? • Spatial Barriers use distance to prevent pollen movement – Studies need to be done to determine the spatial distances on a crop by crop basis 9-13-2002 Purdue 29 What Can Be Done? • Temporal barriers take advantage of known flowering patterns and sexually isolate the crop from the crops around it. – Quite effective – Used by Breeders 9-13-2002 Purdue 30 What Do I Recommend? • Four of the five barriers should be used to prevent or eliminate pollen movement. – Management intensive – Assumes no human errors (bad assumption) 9-13-2002 Purdue 31 What Does Aphis Require • Transgenic corn must be planted at least 1 mile from corn seed production • Corn from previous season in a 0.25 mile radius must be harvested and removed before planting • The land within 25 feet of the transgenic plant area must be fallow • No other corn plants within a radius of 0.5 mile of transgenic plants • Transgenic corn must be planted no less that 21 days before or 21 days after planting dates of corn in a zone of 0.5 to 1 mile of transgenic plants • Buffers reduce distances and times 9-13-2002 Purdue 32 Other Issues • Identity preservation for zero tolerance – Theoretically possible – practically difficult – Costly if areas of production are widely separated – Humans make mistakes • Misappropriation • Spillage 9-13-2002 Purdue 33 Other Recommendations • Both DNA and Protein tests of the gene and the gene product should be available before testing • Genes should be constructed so they are traceable to the lab of origin 9-13-2002 Purdue 34 Conclusion Gene Flow Happens 9-13-2002 Purdue 35