Bioassay and Characterization of Several Palmer Amaranth (Amaranthus palmeri) Biotypes with Varying Tolerances to Glyphosate

Abstract

The wide distribution of Palmer amaranth (Amaranthus palmeri) in the southern US became a serious weed control problem prior to the extensive use of glyphosate-resistant crops. Currently glyphosate-resistant populations of Palmer amaranth occur in many areas of this geographic region creating an even more serious threat to crop production. Investigations were undertaken using four biotypes (one glyphosate-sensitive, one resistant from Georgia and two of unknown tolerance from Mississippi) of Palmer amaranth to assess bioassay techniques for the rapid detection and level of resistance in populations of this weed. These plants were characterized with respect to chlorophyll, betalain, and protein levels and immunological responses to an antibody of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) the target site of glyphosate. Only slight differences were found in four biotypes grown under greenhouse conditions regarding extractable soluble protein and chlorophyll content, but one biotype was found to be devoid of the red pigment, betalain. Measurement of early growth (seedling shoot elongation) of seedlings was a useful detection tool to determine glyphosate resistance. A leaf disc bioassay (using visual ratings and/or chlorophyll analysis) and an assay for shikimate accumulation were effective methods for determining herbicide resistance levels. The two unknown biotypes were found to be resistant to this herbicide. Some differences were found in the protein profiles of the biotypes, and western blots demonstrated a weak labeling of antibody in the glyphosate-sensitive biotype, whereas strong labeling occurred in the resistant plants. This latter point supports research by others, that increased copy number of the EPSPS gene (and increased EPSPS protein levels) is the resistance mechanism in this species. Results indicate the utility of certain bioassays for the determination of resistance and provide useful comparative information on the levels of inherent constituents among closely related plants.

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R. Hoagland, R. Jordan and N. Teaster, "Bioassay and Characterization of Several Palmer Amaranth (Amaranthus palmeri) Biotypes with Varying Tolerances to Glyphosate," American Journal of Plant Sciences, Vol. 4 No. 5, 2013, pp. 1029-1037. doi: 10.4236/ajps.2013.45127.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Heap, “International Survey of Herbicide-Resistant Weeds,” 2013. http://www.weedscience.org/in.asp
[2] Anonymous, “New Report Reveals Dramatic Rise in Pesticide Use on Genetically Engineered (GE) Crops Due to the Spread of Resistant Weeds,” 2009. http://wwwprnewswire.com/news-release
[3] B. G. Young, “Changes in Herbicide Use Patterns and Production Practices Resulting from Glyphosate-Resistant Crops,” Weed Technology, Vol. 20, No. 2, 2006, pp. 301- 307. doi:10.1614/WT-04-189.1
[4] L. D. Bradshaw, S. R. Padgette, S. L. Kimball and B. H. Wells, “Perspectives on Glyphosate Resistance,” Weed Technology, Vol. 11, No. 1, 1997, pp. 189-198.
[5] T. M. Webster, “Weed Survey-Southern States: Broadleaf Crops Subsection,” Proceedings of the Southern Weed Science Society, Vol. 58, 2005, pp. 291-294.
[6] S. Culpepper, T. L. Grey, W. K. Vencill, J. M. Kichler, T. M. Webster, S. M. Brown, A. C. York, J. W. Davis and W. W. Hanna, “Glyphosate-Resistant Palmer Amaranth (Amaranthus palmeri) Confirmed in Georgia,” Weed Science, Vol. 54, No. 4, 2006, pp. 620-626. doi:10.1614/WS-06-001R.1
[7] R. C. Scott, L. E. Steckel, K. L. Smith, T. Mueller, L. R. Oliver and J. Norsworthy, “Glyphosate-Resistant Palmer Amaranth in the Southeastern United States,” In: W. K. Vencill, Ed., Proceedings of the 60th Southern Weed Science Society, Nashville, 22-24 January 2007.
[8] C. York, J. R. Whitaker, A. S. Culpepper and C. L. Main, “Glyphosate-Resistant Palmer Amaranth in the Southeastern United States,” Proceedings of the Southern Weed Science Society, Vol. 60, 2007, p. 225.
[9] N. Bensch, M. J. Horak and D. Peterson, “Interference of Redroot Pigweed (Amaranthus retroflexus), Palmer Amaranth (A. palmeri), and Common Waterhemp (A. rudis) in Soybean,” Weed Science, Vol. 51, No. 1, 2003, pp. 37-43. doi:10.1614/0043-1745(2003)051[0037:IORPAR]2.0.CO;2
[10] A. Sellers, R. J. Smeda, W. G. Johnson, A. J. Kendig and M. R. Ellersieck, “Comparative Growth of Six Amaranthus Species in Missouri,” Weed Science, Vol. 51, No. 3, 2003, pp. 329-333. doi:10.1614/0043-1745(2003)051[0329:CGOSAS]2.0.CO;2
[11] M. J. Horak and T. M. Loughin, “Growth Analysis of Four Amaranthus Species,” Weed Science, Vol. 48, No. 3, 2000, pp. 347-355. doi:10.1614/0043-1745(2000)048[0347:GAOFAS]2.0.CO;2
[12] W. MacRae, A. S. Culpepper, T. M. Webster and J. M. Kichler, “The Effect of Glyphosate-Resistant Palmer Amaranth Density and Time of Establishment on Yield of Cotton,” Proceedings of the Southern Weed Science Society, Vol. 60, 2007, p. 228.
[13] D. T. Smith, R. V. Baker and G. L. Steele, “Palmer Amaranth (Amaranthus palmeri) Impacts on Yield, Harvesting, and Ginning in Dryland Cotton,” Weed Technology, Vol. 14, No. 1, 2000, pp. 122-126. doi:10.1614/0890-037X(2000)014[0122:PAAPIO]2.0.CO;2
[14] S. Culpepper, A. W. MacRae, A. C. York and J. Kichler, “Managing Glyphosate-Resistant Palmer Amaranth in Conventional and Strip-till Roundup Ready Cotton,” Proceedings of the Southern Weed Science Society, Vol. 61, 2008, p. 62.
[15] A. S. Culpepper, A. C. York and J. Kichler, ‘University of Georgia Herbicide Programs for Controlling Glyphosate-Resistant Palmer Amaranth in 2008 Cotton,” Circular No. 924, Georgia Cooperative Extension Service, Tifton, 2008.
[16] S. Culpepper, A. C. York, J. M. Kichler and A. W. MacRae, “Glyphosate-Resistant Palmer Amaranth Response to Weed Management Programs in Roundup Ready and Liberty Link Cotton,” Proceedings of the Beltwide Cotton Conference, Nashville, 10 January 2008, pp. 1689-1690.
[17] J. R. Whitaker, “Distribution, Biology, and Management of Glyphosate-Resistant Palmer Amaranth in North Carolina,” Ph.D. Dissertation, North Carolina State University, Raleigh, 2009, http://repository.lib.ncsu.edu/ir/bitstream/1840.16/3130/1/etd.pdf
[18] L. M. Sosnoskie, J. M. Kichler, R. D. Wallace and A. S. Culpepper, “Multiple Resistance in Palmer Amaranth to Glyphosate and Pyrithiobac Confirmed in Georgia,” Weed Science, Vol. 59, No. 3, 2011, pp. 321-325. doi:10.1614/WS-D-10-00132.1
[19] H. C. Steinrücken and N. Amrhein, “The Herbicide Glyphosate is a Potent Inhibitor of 5-Enolpyruvyl-Shikimic Acid-3-Phosphate Synthase,” Biochemical and Biophysical Research Communications, Vol. 94, No. 4, 1980, pp. 1207-1212. doi:10.1016/0006-291X(80)90547-1
[20] J. E. Franz, M. K. Mao and J. A. Sikorski, “Glyphosate’s Molecular Mode of Action,” In: Glyphosate: A Unique Global Herbicide, American Chemical Society, Washington DC, 1997, pp. 521-615.
[21] K. J. Gruys and J. A. Sikorski, “Inhibitors of Tryptophan, Phenylalanine and Tyrosine Biosynthesis as Herbicides,” In: B. K. Singh, Ed., Plant Amino Acids: Biochemistry and Biotechnology, CRC Press, Boca Raton, 1999, pp. 357-384.
[22] E. Haslam, “Shikimic Acid: Metabolism and Metabolites,” John Wiley Sons, Chichester, 1993, 400 Pages.
[23] K. M. Herrmann, “The Shikimate Pathway: Early Steps in the Biosynthesis of Aromatic Compounds,” Plant Cell, Vol. 7, No. 7, 1995, pp. 907-919.
[24] K. M. Herrmann, “The Shikimate Pathway as an Entry to Aromatic Secondary Metabolism,” Plant Physiology, Vol. 107, No. 1, 1995, pp. 7-12. doi:10.1104/pp.107.1.7
[25] N. Amrhein, B. Deus, P. Gehrke and H. C. Steinrüchen, “The Site of the Inhibition of the Shikimate Pathway by Glyphosate. II. Interference of Glyphosate with Chorismate Formation in Vivo and in Vitro,” Plant Physiology, Vol. 66, No. 5, 1980, pp. 830-834. doi:10.1104/pp.66.5.830
[26] T. A. Gaines, W. Zhang, D. Wang, B. Bukun, S. T. Chisholm, D. L. Shaner, S. J. Nissen, W. L. Patzoldt, P. J. Tranel, A. S. Culpepper, T. L. Grey, T. M. Webster, W. K. Vencill, R. D. Sammons, J. Jiang, C. Preston, J. E. Leach and P. Westra, “Gene Amplification Confers Glyphosate Resistance in Amaranthus palmeri,” Proceedings of the National Academy of Sciences, Vol. 107, No. 3, 2010, pp. 1029-1034. doi:10.1073/pnas.0906649107
[27] L. M. Sosnoskie, T. M. Webster, J. M. Kichler, A. W. MacRae and A. S. Culpepper, “Preliminary Estimates of Glyphosate-Resistant Amaranthus palmeri Pollen Dispersal Distances,” Proceedings of the Beltwide Cotton Conference, New Orleans, 11 January 2007, p. 1228.
[28] J. D. Barnes, L. Balaguer, E. Manriques, S. Elvira and A. W. Davison, “A Reappraisal of the Use of DMSO for the Extraction and Determination of Chlorophylls a and b in Lichens and Higher Plants,” Environmental and Experimental Botany, Vol. 32, No. 2, 1992, pp. 85-100. doi:10.1016/0098-8472(92)90034-Y
[29] J. D. Hiscox and G. F. Israelstam. “A Method for the Extraction of Chlorophyll from Leaf Tissue without Maceration,” Canadian Journal of Botany, Vol. 57, No. 12, 1979, pp. 1332-1334. doi:10.1139/b79-163
[30] R. E. Hoagland, “Effects of Glyphosate on Metabolism of Phenolic Compounds: VI. Effects of Glyphosine and Glyphosate Metabolites on Phenylalanine Ammonialyase Activity, Growth and Protein, Chlorophyll and Anthocyanin Levels in Soybean (Glycine max) Seedlings,” Weed Science, Vol. 28, No. 4, 1980, pp. 393-400.
[31] D. L. Shaner, T. Nadler-Hassar, W. B. Henry and C. H. Koger, “A Rapid in Vivo Shikimate Accumulation Assay with Excised Leaf Discs,” Weed Science, Vol. 53, No. 6, 2005, pp. 769-774. doi:10.1614/WS-05-009R.1
[32] Y. Tanaka, N. Sasaki and A. Ohmiya, “Biosynthesis of Plant Pigments: Anthocyanins, Betalains and Carotenoids,” The Plant Journal, Vol. 54, No. 4, 2008, pp. 733-749. doi:10.1111/j.1365-313X.2008.03447.x
[33] H. J. Beckie, I. M. Heap, R. J. Smeda and L. M. Hall, “Screening for Herbicide Resistance in Weeds,” Weed Technology, Vol. 14, No. 2, 2000, pp. 428-445. doi:10.1614/0890-037X(2000)014[0428:SFHRIW]2.0.CO;2
[34] R. E. Hoagland and R. D. Williams, “Bioassays: Useful Tools for the Study of Allelopathy,” In: F. Macias, J. Galindo, J., Molinillo and H. Cutler, Eds., Allelopathy, CRC Press, 2004, pp. 315-351.
[35] P. Boger and G. Sandmann, “Target Assays for Modern Herbicides and Related Phytotoxic Compounds,” Lewis Publishers, Boca Raton, 1993, 299 pages.
[36] J. C. Streibig, “Herbicide Bioassay,” Weed Research, Vol. 28, No. 6, 1988, pp. 479-484. doi:10.1111/j.1365-3180.1988.tb00831.x
[37] H. Hollander-Czytko and N. Amrhein, “5-EnolpyruvylShikimate 3-Phosphate Synthase, the Target Enzyme of the Herbicide Glyphosate, Is Synthesized as a Precursor in a Higher Plant,” Plant Physiology, Vol. 83, No. 2, 1987, pp. 229-231. doi:10.1104/pp.83.2.229
[38] M. Jasieniuk, A. L. Brule-Babel and I. N. Morrison, “The Evolution and Genetics of Herbicide Resistance in Weeds,” Weed Science, Vol. 44, No. 1, 1996, pp. 176-193.

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