Mutations and amplification of EPSPS gene confer resistance to glyphosate in goosegrass (Eleusine indica)
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Field-evolved resistance of goosegrass to glyphosate is due to double or single mutation in EPSPS , or amplification of EPSPS leads to increased transcription and protein levels.
Glyphosate has been used widely in the south of China. The high selection pressure from glyphosate use has led to the evolution of resistance to glyphosate in weeds. We investigated the molecular mechanisms of three recently discovered glyphosate-resistant Eleusine indica populations (R1, R2 and R3). The results showed that R1 and R2 had double Thr102Ile and Pro106Ser mutation and a single mutation of Pro106Leu in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene, respectively. Escherichia coli containing the mutated EPSPS genes was tolerant to glyphosate. EPSPS activity in R1 and R2 plants was higher than in the sensitive plants. There was no amino acid substitution in EPSPS gene in R3. However, expression of EPSPS in R3 plants was higher than in glyphosate-susceptible (S) population (13.8-fold) after glyphosate treatment. EPSPS enzyme activity in both R3 and S plants was inhibited by glyphosate, while shikimate accumulation in R3 was significantly lower than for the S population. Further analysis revealed that the genome of R3 contained 28.3-fold more copies of the EPSPS gene than that of susceptible population. EPSPS expression was positively correlated with copy number of EPSPS. In conclusion, mutation of the EPSPS gene and increased EPSPS expression are part of the molecular mechanisms of resistance to glyphosate in Eleusine indica.
KeywordsGlyphosate Resistance EPSPS Eleusine indica Mechanism
Polymerase chain reaction
This research was funded by Special Fund for Agro-scientific Research in the Public Interest (201303022). The authors thank the anonymous reviewers for improving the language of the manuscript.
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
- Funke T, Yang Y, Han H, Healy-Fried M, Olesen S, Becker A, Schönbrunn E (2009) Structural basis of glyphosate resistance resulting from the double mutation Thr97 → Ile and Pro101 → Ser in 5-enolpyruvylshikimate-3-phosphate synthase from Escherichia coli. J Biolog Chem 284(15):9854–9860CrossRefGoogle Scholar
- Ge X, d’Avignon DA, Ackerman JJ, Collavo A, Sattin M, Ostrander EL, Hall EL, Sammons RD, Preston C (2012) Vacuolar glyphosate-sequestration correlates with glyphosate resistance in ryegrass (Lolium spp.) from Australia, south America, and Europe: a 31P NMR investigation. J Agri Food Chem 60(5):1243–1250CrossRefGoogle Scholar
- González-Torralva F, Gil-Humanes J, Barro F, Brants I, De Prado R (2012) Target site mutation and reduced translocation are present in a glyphosate-resistant Lolium multiflorum Lam. biotype from Spain. J Agri Food Chem 58:16–22Google Scholar
- Heap I (2015) The international survey of herbicide resistant weeds. www.weedscience.org. Accessed 15 Mar 2015
- Jasieniuk M, Ahmad R, Sherwood AM, Firestone JL, Perez-Jones A, Lanini WT, Mallory-Smith C, Stednick Z (2008) Glyphosate-resistant Italian ryegrass (Lolium multiflorum) in California: distribution, response to glyphosate, and molecular evidence for an altered target enzyme. Weed Sci 56(4):496–502CrossRefGoogle Scholar
- Kahrizi D, Salmanian AH, Afshari A, Moieni A, Mousavi A (2007) Simultaneous substitution of Gly96 to Ala and Ala183 to Thr in 5-enolpyruvylshikimate-3-phosphate synthase gene of E. coli (k12) and transformation of rapeseed (Brassica napus L.) in order to make tolerance to glyphosate. Plant Cell Rep 26:95–104CrossRefPubMedGoogle Scholar
- Lebrun M, Sailland A, Freyssinet G and Degryse E (2003) Mutated 5-enolpyruvylshikimate-3-phosphate synthase, gene coding for said protein and transformed plants containing said gene. US 6,566,587, Bayer CropScience SA, pp 1–17Google Scholar
- Nandula VK, Wright AA, Bond JA, Ray JD, Eubank TW, Molin WT (2014) EPSPS amplification in glyphosate-resistant spiny amaranth (Amaranthus spinosus): a case of gene transfer via interspecific hybridization from glyphosate-resistant Palmer amaranth (Amaranthus palmeri). Pest Manag Sci 70(12):1902–1909CrossRefPubMedGoogle Scholar
- Pratley J, Urwin N, Stanton R, Baines P, Broster J, Cullis K, Schafer D, Bohn J, Krueger R (1999) Resistance to glyphosate in Lolium rigidum. I. Bioevaluation. Weed Sci 47:405–411Google Scholar
- Seefeldt S, Jensen J, Fuerst E (1995) Log-logistic analysis of herbicide dose-response relationship. Weed Technol 9:218–227Google Scholar
- Yang CH, Tian XS, Feng L, Yue MF (2012) Resistance of Eleusine indica Gaertn to glyphosate. Scientia Agricultura Sinica 45(10):2093–2098Google Scholar