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Rapid Identification and Molecular Characterization of Phytoene Desaturase Mutations in Fluridone-Resistant Hydrilla (Hydrilla verticillata)

  • Lori K. Benoit (a1) and Donald H. Les (a1)


Florida hydrilla populations have shown an alarming increase in resistance to fluridone, an herbicide used extensively for controlling invasive US hydrilla populations. A rapid PCR and sequencing method was developed to identify and screen hydrilla genomic DNA for three previously identified phytoene desaturase (pds) gene mutations that confer resistance to fluridone. Ninety hydrilla accessions were screened for fluridone resistant genotypes including 46 accessions from the US and 44 accessions from 15 other countries. In Florida, hydrilla from five of nine sites tested was heterozygous for wild-type and herbicide-resistant alleles. Additionally, a new resistant population was identified from Lake Seminole in Georgia, the first genetically confirmed strain of resistant hydrilla outside of Florida. All resistance-conferring mutations were located on the same homologous haplotype of US dioecious hydrilla. All other hydrilla samples tested possessed only wild type alleles, including monoecious strains that had been exposed to fluridone. Management implications are discussed.


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Arias, R. S., Netherland, M. D., Scheffler, B. E., Puri, A., and Dayan, F. E. 2005. Molecular evolution of herbicide resistance to phytoene desaturase inhibitors in Hydrilla verticillata and its potential use to generate herbicide-resistant crops. Pest. Manag. Sci. 61:258268.
Asplund, T. 2007. Hydrilla in Wisconsin. Aquatic Plant News. 86:6.
Bartley, G. E., Viitanen, P. V., Pecker, I., Chamovitz, D., Hirschberg, J., and Scolnik, P. A. 1991. Molecular cloning and expression in photosynthetic bacteria of a soybean cDNA coding for phytoene desaturase, an enzyme of the carotenoid biosynthesis pathway. Proc. Natl. Acad. Sci. U. S. A. 88:65326536.
Beckie, H. J. 2006. Herbicide-resistant weeds: Management tactics and practices. Weed Tech. 20:793814.
Benoit, L. K. 2011. Cryptic speciation, genetic diversity and herbicide resistance in the invasive aquatic plant Hydrilla verticillata (L.f.) Royle (Hydrocharitaceae). PhD dissertation, Storrs, CT University of Connecticut. 117 p.
Brooker, M. P. and Edwards, R. W. 1975. Aquatic herbicides and the control of water weeds. Water Res. 9:115.
Chamovitz, D., Sandmann, G., and Hirschberg, J. 1993. Molecular and biochemical characterization of herbicide-resistant mutants of cyanobacteria reveals that phytoene desaturation is a rate-limiting step in carotenoids biosynthesis. J. Biol. Chem. 268:1734817353.
Cook, C. D. K. and Luond, R. 1982. A revision of the genus Hydrilla (Hydrocharitaceae). Aquat. Bot. 13:485504.
Darmency, H. and Gasquez, J. 1990. Fate of herbicide resistance genes in weeds. Pages 354363 in Green, M., Moberg, W., and LeBaron, H., eds. Managing Resistance to Agrochemicals. Washington, DC American Chemical Society.
Dayan, F. E. and Netherland, M. D. 2005. Hydrilla, the perfect aquatic weed, becomes more noxious than ever. Outlooks Pest Man. 16:277282.
Délye, C., Michel, S., Bérard, A., Chauvel, B., Brunel, D., Guillemin, J-P., Dessaint, F., and Le Corre, V. 2010. Geographical variation in resistance to acetyl-coenzyme A carboxylase-inhibiting herbicides across the range of the arable weed Alopecurus myosuroides (black grass). New Phytol. 186:10051017.
Diggle, A. J. and Neve, P. 1998. The population dynamics and genetics of herbicide resistance – a modeling approach. Pages 6199 in Powles, S. and Shaner, D., eds. Herbicide Resistance and World Grains. Boca Raton, FL CRC.
Doong, R. L., MacDonald, G. E., and Shilling, D. G. 1993. Effect of fluridone on chlorophyll, carotenoids and anthocyanin content of hydrilla. J. Aquat. Plant Manage. 31:5559.
Doyle, J. J. and Doyle, J. L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19:1115.
EDDMapS. 2012. Early Detection & Distribution Mapping System. The University of Georgia - Center for Invasive Species and Ecosystem Health. Available online at; last Accessed August 6, 2012.
Fox, A. M., Haller, W. T., and Shilling, D. G. 1994. Use of fluridone for hydrilla management in the Withlacoochee River, Florida. J. Aquat. Plant Manage. 32:4755.
Hamelink, J. L., Buckler, D. R., Mayer, F. L., Palawski, D. U., and Sanders, H. O. 1986. Toxicity of fluridone to aquatic invertebrates and fish. Environ. Toxicol. Chem. 5:8794.
Heap, I. 2012. The International survey of herbicide resistant weeds. Accessed August 7, 2012.
Hoyer, M. V., Netherland, M. D., Allen, M. S., and Canfield, D. E. Jr. 2005. Hydrilla management in Florida: A summary and discussion of issues identified by professionals with future management recommendations. Accessed May 22, 2012.
Jasieniuk, M., Brûlé-Babel, A. L., and Morrison, I. N. 1996. The evolution and genetics of herbicide resistance in weeds. Weed Sci. 44:176193.
Keohavong, P. and Thilly, W. G. 1989. Fidelity of DNA polymerases in DNA amplification. Proc. Natl. Acad. Sci. USA. 86:92539257.
Koschnick, T. J., Haller, W. T., and Netherland, M. D. 2006. Aquatic plant resistance to herbicides. Aquatics 28:49.
Kovalchuk, I., Kovalchuk, O., and Hohn, B. 2000. Genome-wide variation of the somatic mutation frequency in transgenic plants. EMBO J. 19:44324438.
Les, D. H., Mehrhoff, L. J., Cleland, M. A., and Gabel, J. D. 1997. Hydrilla verticillata (Hydrocharitaceae) in Connecticut. J. Aquat. Plant Manage. 35:1014.
Maddison, D. R. and Maddison, W. P. 2000. MacClade 4: Analysis of phylogeny and character evolution. Sinauer, Sunderland, MA. 492 p.
Madeira, P. T., Jacono, C. C., and Van, T. K. 2000. Monitoring hydrilla using two RAPD procedures and the nonindigenous aquatic species database. J. Aquat. Plant Manage. 38:3340.
Madeira, P. T., Van, T. K., and Center, T. D. 2004. An improved molecular tool for distinguishing monoecious and dioecious hydrilla. J. Aquat. Plant Manage. 42:2832.
Michel, A., Arias, R. S., Scheffler, B. E., Duke, S. O., Netherland, M., and Dayan, F. E. 2004. Somatic mutation-mediated evolution of herbicide resistance in the nonindigenous invasive plant hydrilla (Hydrilla verticillata). Mol. Ecol. 13:32293237.
Murphy, K. J. 1988. Aquatic weed problems and their management: a review I. The worldwide scale of the aquatic weed problem. Crop Prot. 7:232248.
Netherland, M. D., Getsinger, K. D., and Skogerboe, J. D. 1997. Mesocosm evaluation of the selective potential of fluridone. J. Aquat. Plant Manage. 35:4150.
Pons, L. 2005. Pegging a troublesome change in hydrilla. Agric. Res. 53:1416.
Puri, A., MacDonald, G. E., Singh, M., and Haller, W. T. 2006. Phytoene and β-carotene response of fluridone-susceptible and -resistant hydrilla (Hydrilla verticillata) biotypes to fluridone. Weed Sci. 54:995999.
Puri, A., MacDonald, G. E., Altpeter, F., and Haller, W. T. 2007. Mutations in phytoene desaturase gene in fluridone-resistant Hydrilla (Hydrilla verticillata) biotypes in Florida. Weed Sci. 55:412420.
Rogstad, S. H. 1992. Saturated NaCl-CTAB solution as a means of field preservation of leaves for DNA analyses. Taxon 41:701708.
Saari, L. L., Cotterman, J. C., and Thill, D. C. 1994. Resistance to acetolactase synthase inhibiting herbicides. Pages 83140 in Powles, S. B. and Holtum, J. A. M., eds. Herbicide Resistance in Plants: Biology and Biochemistry. Boca Raton, FL Lewis.
Shivrain, V. K., Burgos, N. R., Sales, M. A., and Kuk, Y. I. 2010. Polymorphisms in the ALS gene of weedy rice (Oryza sativa L.) accessions with different tolerance to imazethapyr. Crop Prot. 29:336341.
Speksnijder, A. G. C. L., Kowalchuk, G. A., De Jong, S., Kline, E., Stephen, J. R., and Laanbroek, H. J. 2001. Microvariation artifacts introduced by PCR and cloning of closely related 16S rRNA gene sequences. Appl. Environ. Microbiol. 67:469472.
Steward, K. K., Van, T. K., Carter, V., and Pieterse, A. H. 1984. Hydrilla invades Washington, D.C. and the Potomac. Amer. J. Bot. 1971:162163.
Swofford, D. L. 2002. PAUP*, Phylogenetic Analysis Using Parsimony (*and other methods), version 4.0. Sunderland, MA Sinauer. 142 p.
Vaillant, I. and Paszkowski, J. 2007. Role of histone and DNA methylation in gene regulation. Curr. Opin. Plant Biol. 10 528533.



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