|
|
|
|
|
Overexpression of G10-EPSPS in soybean provides high glyphosate tolerance |
XIAO Pei-ying, LIU Yi, CAO Yue-ping |
School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, P.R.China |
|
|
|
|
Abstract Glyphosate is a highly efficient, broad-spectrum nonspecific herbicide that inhibits the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)-mediated pathway of shikimic acid. The screening of glyphosate-resistant EPSPS gene is a major means for the development of new genetically modified glyphosate-resistant transgenic crop. Currently, the main commercialized glyphosate-resistant soybean contains glyphosate-resistant gene CP4-EPSPS. In this study, a G10-EPSPS gene was reported providing glyphosate resistance in Zhongdou 32. Here, G10-EPSPS gene was introduced into soybeans through Agrobacterium-mediated soybean cotyledon node. PCR, Southern blotting, semi-quantitative RT-PCR, qRT-PCR, and Western blotting were used, and the results revealed that G10-EPSPS had been integrated into the soybean genome and could be expressed steadily at both mRNA and protein levels. In addition, glyphosate resistance analysis showed that the growth of transgenic soybean had not been affected by concentrations of 900 and 2 700 g a.e. ha–1 of glyphosate. All the results indicated that G10-EPSPS could provide high glyphosate resistance in soybeans and be applied in production of glyphosate-resistant soybean.
|
|
Received: 09 April 2018
Accepted:
|
| Fund: This research was financially supported by the National Transgenic Major Program, China (2016ZX08004001-04). |
|
Corresponding Authors:
Correspondence CAO Yue-ping, Tel: +86-21-34206940, E-mail: yuepingcao@sjtu.edu.cn
|
Cite this article:
XIAO Pei-ying, LIU Yi, CAO Yue-ping.
2019.
Overexpression of G10-EPSPS in soybean provides high glyphosate tolerance . Journal of Integrative Agriculture, 18(8): 1851-1858.
|
Baerson S R, Rodriguez D J, Tran M, Feng Y, Biest N A, Dill G M. 2002. Glyphosate-resistant goosegrass. Identification of a mutation in the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase. Plant Physiology, 129, 1265–1275.
Barry G F, Kishore G M, Padgette S R, Stallings W C. 1997. Glyphosate-Tolerant 5-Enolpyruvylshikimate-3-Phosphate Synthases. United States Patent. US19940306063, 1994-09-13.
Bohm G M B, Rombaldi C V, Genovese M I, Castilhos D, Alves B J R, Rumjanek N G. 2014. Glyphosate effects on yield, nitrogen fixation, and seed quality in glyphosate-resistant soybean. Crop Science, 54, 1737.
Bonny S. 2011. Herbicide-tolerant transgenic soybean over 15 years of cultivation: Pesticide use, weed resistance, and some economic issues: The case of the USA. Sustainability, 3, 1302–1322.
Comai L, Facciotti D, Hiatt W R, Thompson G, Rose R E, Stalker D M. 1985. Expression in plants of a mutant aroA gene from Salmonella typhimurium confers tolerance to glyphosate. Nature, 317, 741–744.
Comai L, Sen L C, Stalker D M. 1983. An altered aroA gene product confers resistance to the herbicide glyphosate. Science, 221, 370.
Dellacioppa G, Bauer S C, Taylor M L, Rochester D E, Klein B K, Shah D H, Fraley R T, Kishore G M. 1987. Targeting a herbicide-resistant enzyme from Escherichia coli to chloroplasts of higher plants. Nature Biotechnology, 5, 579–584.
Dill G M, Cajacob C A, Padgette S R. 2008. Glyphosate-resistant crops: Adoption, use and future considerations. Pest Management Science, 64, 326.
Ding W, Reddy K N, Zablotowicz R M, Bellaloui N, Arnold B H. 2011. Physiological responses of glyphosate-resistant and glyphosate-sensitive soybean to aminomethylphosphonic acid, a metabolite of glyphosate. Chemosphere, 83, 593.
Duke S O. 1995. Herbicide-resistant crops: Agricultural, economic, environmental, regulatory, and technological aspects. Journal of Southeast University, 31, 259–265.
Edwards R, Hannah M. 2014. Focus on weed control. Plant Physiology, 166, 1087–1089.
Endo I, Hiraiwa K, Koide T. 2010. Effect of Weeding of Naturalized Morningglories in Soybean Field by Inter-Ridge Herbicide Spraying. Research Bulletin of the Aichi Agricultural Research Center.
Funke T, Han H, Healy-Fried M L, Fischer M, Schönbrunn E. 2006. Molecular basis for the herbicide resistance of roundup ready crops. Proceedings of the National Academy of Sciences of the United States of America, 103, 13010.
Funke T, Yang Y, Han H, Healy-Fried M, Olesen S, Becker A, Schonbrunn 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. Journal of Biological Chemistry, 284, 9854–9860.
Gao Y, Tao B, Qiu L, Jin L, Wu J. 2014. Role of physiological mechanisms and EPSPS gene expression in glyphosate resistance in wild soybeans (Glycine soja). Pesticide Biochemistry and Physiology, 109, 6–11.
Gocal G F W, Knuth M E, Beetham P R. 2012. EPSPS Mutants. United States Patent. US8268622 B2.
Gomes M P, Smedbol E, Chalifour A, Hénault-Ethier L, Labrecque M, Lepage L, Lucotte M, Juneau P. 2014. Alteration of plant physiology by glyphosate and its by-product aminomethylphosphonic acid: An overview. Journal of Experimental Botany, 65, 4691–4703.
Grossbard E, Atkinson D. 1985. The Herbicide Glyphosate. Butterworths, America.
Guo B, Yong G, Hong H, Jin L, Zhang L, Chang R, Lu W, Lin M, Qiu L. 2015. Co-expression of G2-EPSPS and glyphosate acetyltransferase GAT genes conferring high tolerance to glyphosate in soybean. Frontiers in Plant Science, 6, 847.
Han H, Vila-Aiub M M, Jalaludin A, Yu Q, Powles S B. 2017. A double EPSPS gene mutation endowing glyphosate resistance shows a remarkably high resistance cost. Plant Cell & Environment, 40, 3031–3042.
Hartman G L, West E D, Herman T K. 2011. Crops that feed the World 2. Soybean - Worldwide production, use, and constraints caused by pathogens and pests. Food Security, 3, 5–17.
Healyfried M L, Funke T, Priestman M A, Han H, Schönbrunn E. 2007. Structural basis of glyphosate tolerance resulting from mutations of Pro101 in Escherichia coli 5-enolpyruvylshikimate-3-phosphate synthase. Journal of Biological Chemistry, 282, 32949–32955.
Heap I. 2014. Global perspective of herbicide-resistant weeds. Pest Management Science, 70, 1306.
Herrmann K M, Weaver L M. 1999. The shikimate pathway. Annual Review of Plant Physiology & Plant Molecular Biology, 50, 473.
Hoagland R E. 2001. Effect of glyphosate on growth, chlorophyll, and nodulation in glyphosate-resistant and susceptible soybean (Glycine max) varieties. Journal of New Seeds, 2, 37–52.
Liu X, He H, Wang Y, Zhu S, Piao X. 2008. Transesterification of soybean oil to biodiesel using CaO as a solid base catalyst. Fuel, 87, 216–221.
Lu L, Han Q, Li L, Zhou L, Liu R, Song Z, Shen Z, Shou H. 2014. Establishment of an efficient transformation protocol for soybean using glyphosate as selective agent and the development of glyphosate-tolerant transgenic soybean lines. Scientia Sinica Vitae, 44, 406–415. (in Chinese)
Padgette S R, Kolacz K H, Delannay X, Re D B, LaVallee, B J, Tinius C N, Rhodes W K, Otero Y I, Barry G F, Eichholtz D A, Peschke V M, Nida D L, Taylor N B, Kishore G M. 1995. Development, identification, and characterization of a glyphosate-tolerant soybean line. Crop Science, 35, 1451–1461.
Paz M M, Martinez J C, Kalvig A B, Fonger T M, Wang K. 2006. Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation. Plant Cell Reports, 25, 248.
Peng R H, Tian Y S, Xiong A S, Zhao W, Fu X Y, Han H J, Chen C, Jin X F, Yao Q H. 2012. A novel 5-enolpyruvylshikimate-3-phosphate synthase from Rahnella aquatilis with significantly reduced glyphosate sensitivity. PLoS ONE, 7, e39579.
Pline W A, Wilcut J W, Duke S O, Edmisten K L, Wells R. 2002. Tolerance and accumulation of shikimic acid in response to glyphosate applications in glyphosate-resistant and nonglyphosate-resistant cotton (Gossypium hirsutum L.). Journal of Agricultural & Food Chemistry, 50, 506.
Priestman M A, Healy M L, Funke T, Becker A, Schönbrunn E. 2005. Molecular basis for the glyphosate-insensitivity of the reaction of 5-enolpyruvylshikimate 3-phosphate synthase with shikimate. FEBS Letters, 579, 5773–5780.
Randall J M. 1997. Defining Weeds of Natural Areas. Springer, New York.
Sauer N J, Narvã E S J, Mozoruk J, Miller R B, Warburg Z J, Woodward M J, Mihiret Y A, Lincoln T A, Segami R E, Sanders S L, Walker K A, Beetham P R, Schöpke C R, Gocal G F W. 2016. Oligonucleotide-mediated genome editing provides precision and function to engineered nucleases and antibiotics in plants. Plant Physiology, 170, 1917–1928.
Schönbrunn E, Eschenburg S, Shuttleworth W A, Schloss J V, Amrhein N, Evans J N S, Kabsch W. 2001. Interaction of the herbicide glyphosate with its target enzyme 5-enolpyruvylshikimate 3-phosphate synthase in atomic detail. Proceedings of the National Academy of Sciences of the United States of America, 98, 1376–1380.
Singh H B, Singh B N, Singh S P, Nautiyal C S. 2010. Solid-state cultivation of Trichoderma harzianum NBRI-1055 for modulating natural antioxidants in soybean seed matrix. Bioresource Technology, 101, 6444–6453.
Southern E. 2006. Southern blotting. Nature Protocols, 1, 518–525.
Stalker D M, Hiatt W R, Comai L. 1985. A single amino acid substitution in the enzyme 5-enolpyruvylshikimate-3-phosphate synthase confers resistance to the herbicide glyphosate. Journal of Biological Chemistry, 260, 4724–4728.
Stewart Jr C N, Via L E. 1993. A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. Biotechniques, 14, 748–750.
Tan S, Evans R, Singh B. 2006. Herbicidal inhibitors of amino acid biosynthesis and herbicide-tolerant crops. Amino Acids, 30, 195–204.
Xiao X, Wu H H, Zhou X H, Sheng X, He J, Shen W B, Zhou G H, Huang M. 2012. The combination of quantitative PCR and Western blot detecting CP4-EPSPS component in roundup ready soy plant tissues and commercial soy-related foodstuffs. Journal of Food Science, 77, C603–C608.
Zhang Z F. 2011. The application of chemical herbicides in forestry projects. Sci-Tech Information Development & Economy, 35, 223–225. (in Chinese)
Zhou M, Xu H, Wei X, Ye Z, Wei L, Gong W, Wang Y, Zhu Z. 2006. Identification of a glyphosate-resistant mutant of rice 5-enolpyruvylshikimate 3-phosphate synthase using a directed evolution strategy. Plant Physiology, 140, 184.
Zobiole L H S, Kremer R J, de Oliveira Jr R S, Constantin J. 2011. Glyphosate affects chlorophyll, nodulation and nutrient accumulation of “second generation” glyphosate-resistant soybean (Glycine max L.). Pesticide Biochemistry & Physiology, 99, 53–60.
Zobiole L H S, Kremer R J, de Oliveira Jr R S, Constantin J. 2012. Glyphosate effects on photosynthesis, nutrient accumulation, and nodulation in glyphosate-resistant soybean. Journal of Plant Nutrition and Soil Science, 175, 319–330.
Zobiole L H S, de Oliveira Jr R S, Kremer R J, Constantin J, Bonato C M, Muniz A S. 2010. Water use efficiency and photosynthesis of glyphosate-resistant soybean as affected by glyphosate. Pesticide Biochemistry & Physiology, 97, 182–193.
|
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
