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Journal of Integrative Agriculture  2015, Vol. 14 Issue (5): 909-918    DOI: 10.1016/S2095-3119(14)60890-X
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Investigating the mechanisms of glyphosate resistance in goosegrass (Eleusine indica) population from South China
 ZHANG Chun, FENG Li, HE Ting-ting, YANG Cai-hong, CHEN Guo-qi, TIAN Xing-shan
Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Institute of Plant Protection, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, P.R.China
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摘要  Glyphosate has been used worldwide for nearly 40 years, and 30 types of resistant weeds have been reported. Glyphosate is mass-produced and widely used in China, but few studies and reports on glyphosate-resistant weeds and resistance mechanisms exist. Previous studies found a goosegrass species with high glyphosate resistance from orchards in South China and its glyphosate resistant mechanism was described in this study. The cDNA of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19), the target enzyme of glyphosate, was cloned from the glyphosate-resistant and -susceptible goosegrass, respectively, and referred as EPSPS-R and EPSPS-S. The Pro106 residue was known to be involved in the glyphosate resistance in most goosegrass populations. However, sequence analysis did not find the mutation at the Pro106 residue in the R biotype EPSPS amino acid sequence. The residue 133 and 382 was mutated in the R biotype EPSPS amino acid sequence instead, but it did not affect the EPSPS-S and EPSPS-R genes sensitivities to glyphosate. RT-PCR and Western blot analyses suggested that EPSPS mRNA and protein are mainly present in the shoot tissues both in the R and S goosegrass biotypes. The EPSPS-R rapidly responds to the glyphosate in R-biotype goosegrass and the induced expression was detected at 12 h post glyphosate treatment. The mRNA and protein expression of EPSPS-R increased constantly as the increasing concentration of glyphosate. However, the expression of the EPSPS-S was not induced significantly by glyphosate in the S goosegrass biotype. Quantification of real-time PCR results showed that the copy number of the EPSPS in R-biotype goosegrass was 4.7 times higher than that in the S goosegrass biotype. All the results implied that EPSPS gene amplification might mainly caused the glyphosate resistance of a goosegrass population collected from orchards in South China.

Abstract  Glyphosate has been used worldwide for nearly 40 years, and 30 types of resistant weeds have been reported. Glyphosate is mass-produced and widely used in China, but few studies and reports on glyphosate-resistant weeds and resistance mechanisms exist. Previous studies found a goosegrass species with high glyphosate resistance from orchards in South China and its glyphosate resistant mechanism was described in this study. The cDNA of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19), the target enzyme of glyphosate, was cloned from the glyphosate-resistant and -susceptible goosegrass, respectively, and referred as EPSPS-R and EPSPS-S. The Pro106 residue was known to be involved in the glyphosate resistance in most goosegrass populations. However, sequence analysis did not find the mutation at the Pro106 residue in the R biotype EPSPS amino acid sequence. The residue 133 and 382 was mutated in the R biotype EPSPS amino acid sequence instead, but it did not affect the EPSPS-S and EPSPS-R genes sensitivities to glyphosate. RT-PCR and Western blot analyses suggested that EPSPS mRNA and protein are mainly present in the shoot tissues both in the R and S goosegrass biotypes. The EPSPS-R rapidly responds to the glyphosate in R-biotype goosegrass and the induced expression was detected at 12 h post glyphosate treatment. The mRNA and protein expression of EPSPS-R increased constantly as the increasing concentration of glyphosate. However, the expression of the EPSPS-S was not induced significantly by glyphosate in the S goosegrass biotype. Quantification of real-time PCR results showed that the copy number of the EPSPS in R-biotype goosegrass was 4.7 times higher than that in the S goosegrass biotype. All the results implied that EPSPS gene amplification might mainly caused the glyphosate resistance of a goosegrass population collected from orchards in South China.
Keywords:  5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)       gene amplification       glyphosate resistance       Eleusine indica       over-expression  
Received: 22 May 2014   Accepted:
Fund: 

The present study was supported by the National Natural Science Foundation of China (31301683) and the Science and Technology Planning Project of Guangdong Province, China (2012A020100009).

Corresponding Authors:  TIAN Xing-shan, Tel: +86-20-85518286,Fax: +86-20-87561757, E-mail: xstian@tom.com     E-mail:  xstian@tom.com
About author:  ZHANG Chun, E-mail: zhangchun_0726@163.com;

Cite this article: 

ZHANG Chun, FENG Li, HE Ting-ting, YANG Cai-hong, CHEN Guo-qi, TIAN Xing-shan. 2015. Investigating the mechanisms of glyphosate resistance in goosegrass (Eleusine indica) population from South China. Journal of Integrative Agriculture, 14(5): 909-918.

Baerson S R, Rodriguez D J, Biest N A, Tran M, You J S, KreugerR W, Dill G M, Pratley J E, Gruy K J. 2002a. Investigatingthe mechanism of glyphosate resistance in rigid ryegrass(Lolium ridigum). Weed Science, 50, 721-730

Baerson S R, Rodriguez D J, Tran M, Feng Y M, Biest N A, DillG M. 2002b. Glyphosate-resistant goosegrass. Identificationof a mutation in the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase. Plant Physiology, 129, 1265-1275

Dinelli G, Marotti I, Bonetti A, Minelli M, Catizone P, BarnesJ. 2006. Physiological and molecular insight on themechanisms of resistance to glyphosate in Conyzacanadensis (L.) Cronq. biotypes. Pesticide Biochemistryand Physiology, 86, 30-41

Gaines T A, Wright A A, Molin W T, Lorentz L, Riggins CW, Tranel P J, Beffa R, Westra P, Powles S B. 2013.Identification of genetic elements associated with EPSPSgene amplification. PLOS ONE, 8, e65819.

Gaines T A, Zhang W L, Wang D F, Bukun B, Chisholm S T,Shaner D L, Nissen S J, Patzoldt W L, Tranel P J, CupepperA S, Grey T L, Webster T M, Vencill W K, Sammons RD, Jiang J, Preston C, Leach J E, Westra P. 2010. Geneamplification confers glyphosate resistance in Amaranthuspalmeri. Proceedings of the National Academy of Sciencesof the United States of America, 107, 1029-1034

Ge X, D’Avignon D A, Ackerman J J, Sammons R D. 2010.Rapid vacuolar sequestration: the horseweed glyphosateresistance mechanism. Pest Management Science, 66,345-348

Heap I M. 2014. International survey of herbicide-resistantweeds. [2014-09-27] http://wwwweedscience.org/summary/Species.aspx

Holm L G, Plucknett D L, Pancho J V, Herberger J P. 1977. TheWorld’s Worst Weeds: Distribution and Biology. UniversityPress of Hawaii, Honolulu. pp. 47-53

Huang J L, Silva E N, Shen Z G, Jiang B, Lu H F. 2012. Effectsof glyphosate on photosynthesis, chlorophyll fluorescenceand physicochemical properties of cogongrass (Imperatacylindrical L.). Plant Omics, 5, 177-183

Kaundun S S, Zelaya I A, Dale R P, Lycett A J, Carter P,Sharples K R and McIndoe E. 2008. Importance of thep106s target-site mutation in conferring resistance toglyphosate in a goosegrass (Eleusine indica) populationfrom the Philippines. Weed Science, 56, 637-646

Lee L J, Ngim J. 2000. A first report of glyphosate-resistantgoosegrass (E. indica (L.) Gaertn) in Malaysia. PestManagement Science, 56, 336-339

Liu Y, Cui H L, Huang H J, Wei S H, Zhang C X. 2008.Advances in research on glyphosate-resistant weeds andtheir mechanism. Chinese Journal of Pesticide Science,10, 10-14 (in Chinese)

Lorraine-Colwill D F, Powles S B, Hawkes T R, Hollinshead PH, Warner S A J, Preston C. 2001. Inheritance of evolvedglyphosate resistance in Lolium rigidum. Theoretical andApplied Genetics, 102, 545-550

Lorraine-Colwill D F, Powles S B, Hawkes T R, Hollinshead PH, Warner S A J, Preston C. 2003. Investigations into themechanism of glyphosate resistance in Lolium rigidum.Pesticide Biochemistry and Physiology, 74, 62-72

Ng C H, Wickneswary R, Salmijah S, Teng Y T, Ismail B S. 2004.Glyphosate resistance in Eleusine indica (L.) Gaertn. fromdifferent origins and polymerase chain reaction amplificationof specific alleles. Australian Journal of AgriculturalResearch, 55, 407-414

Powles S B. 2008. Evolved glyphosate-resistant weeds aroundthe world: Lessons to belearnt. Pest Management Science,64, 360-365

Powles S B 2010. Gene amplification delivers glyphosateresistantweed evolution. Proceedings of the NationalAcademy of Sciences of the United States of America,107, 955-956

Powles S B, Lorraine-Colwill D F, Dellow J J, Preston C. 1998.Evolved resistance to glyphosate in rigid ryegrass (Loliumrigidum) in Australia. Weed Science, 46, 604-607

Powles S B, Yu Q. 2010. Evolution in action: Plants resistant toherbicides. Annual Review of Plant Biology, 61, 317-347

Salas R A, Dayan F E, Pan Z, Watson S B, Dickson J W, ScottR C, Burgos N R. 2012. EPSPS gene amplification inglyphosate-resistant Italian ryegrass (Lolium perenne ssp.multiflorum) from Arkansas. Pest Management Scienc68, 1223-1230

Shaner D L, Lindenmeyer R B, Ostlie M H. 2012. What havethe mechanisms of resistance to glyphosate taught us?Pest Management Science, 68, 3-9

Simarmata M, Penner D. 2008. The basis for glyphosateresistance in rigid ryegrass (Lolium rigidum) from California.Weed Science, 56, 181-188

Song X L, Wu J J, Zhang H J, Qiang S. 2011. Occurrenceof glyphosate-resistant horseweed (Conyza canadensis)population in China. Agricultural Sciences in China, 10,1049-1055

Yang C H, Feng L, Yue M F, Tian X S. 2009. Study on SeedGermination Characteristics of Goosegrass (Eleusineindica). Weed Science, 9, 21-24 (in Chinese)

Yang C H, Tian X S, Feng L, Yue M F. 2012. Resistance ofEleusine indica Gaertn to glyphosate. Scentiai AgriculturaSinica, 45, 2093-2098 (in Chinese)

Yuan C I, Chaing M Y, Chen Y M. 2002. Triple mechanisms ofglyphosate-resistance in a naturally occurring glyphosateresistantplant Dicliptera chinensis, Plant Science, 163,543-554

Zhang C, Wu D D, Feng L, Tian X S, Guo A L. 2014, Cloningand functional verification of a target gene of glyphosatefrom Klebsiella pneumoniae S001. Scientia AgriculturaSinica, 457, 80-89 (in Chinese)
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