Please wait a minute...
Journal of Integrative Agriculture  2011, Vol. 10 Issue (9): 1307-1312    DOI: 10.1016/S1671-2927(11)60123-5
GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS Advanced Online Publication | Current Issue | Archive | Adv Search |
Development of Transgenic Glyphosate-Resistant Rice with G6 GeneEncoding 5-Enolpyruvylshikimate-3-Phosphate Synthase
ZHAO Te, LIN Chao-yang , SHEN Zhi-cheng
State Key Laboratory of Rice Biology and Institute of Insect Sciences, Zhejiang University
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  Glyphosate-resistant crops have been a huge economic success for genetic engineering. The creating of new glyphosateresistantplants would increase the available choices for planting and lower the price of genetically modified crop seeds.A novel G6 gene from Pseudomonas putida that encoded 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) waspreviously isolated. The G6 gene was transfected into rice via Agrobacterium-mediated transformation. The transgenicrice obtained was confirmed by PCR, Southern, and Western blots. The lab experiment and field trials further confirmedthat the transgenic rice can survive glyphosate spraying at a dose of 8 g L-1. In contrast, conventional rice was killed ata weed control glyphosate spray dose of 1 g L-1. Altogether, the present study showed that the G6 gene works well in ricein vivo for glyphosate-resistance.

Abstract  Glyphosate-resistant crops have been a huge economic success for genetic engineering. The creating of new glyphosateresistantplants would increase the available choices for planting and lower the price of genetically modified crop seeds.A novel G6 gene from Pseudomonas putida that encoded 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) waspreviously isolated. The G6 gene was transfected into rice via Agrobacterium-mediated transformation. The transgenicrice obtained was confirmed by PCR, Southern, and Western blots. The lab experiment and field trials further confirmedthat the transgenic rice can survive glyphosate spraying at a dose of 8 g L-1. In contrast, conventional rice was killed ata weed control glyphosate spray dose of 1 g L-1. Altogether, the present study showed that the G6 gene works well in ricein vivo for glyphosate-resistance.
Keywords:  transgenic rice      glyphosate-resistance      EPSPS  
Received: 02 June 2011   Accepted:
Corresponding Authors:  Correspondence SHEN Zhi-cheng, Professor, Tel/Fax: +86-571-86971273, E-mail: zcshen@zju.edu.cn     E-mail:  zcshen@zju.edu.cn
About author:  ZHAO Te, Ph D, Mobile: 15838136601, E-mail: tezhao@126.com

Cite this article: 

ZHAO Te, LIN Chao-yang , SHEN Zhi-cheng. 2011. Development of Transgenic Glyphosate-Resistant Rice with G6 GeneEncoding 5-Enolpyruvylshikimate-3-Phosphate Synthase. Journal of Integrative Agriculture, 10(9): 1307-1312.

[1]Battraw M J, Hall T C. 1990. Histochemical analysis of CaMV35S promoter-β-glucuronidase gene expression in transgenicrice plants. Plant Molecular Biology, 15, 527-538.

[2]Cao M X, Huang J Q, Wei Z M, Yao Q H, Wan C Z, Lu J A.2004. Engineering higher yield and herbicide resistance inrice by agrobacterium-mediated multiple gene transformation.Crop Science, 44, 2206-2213.

[3]Christensen A H, Sharrock R A, Quail P H. 1992. Maizepolyubiquitin genes: structure, thermal perturbation ofexpression and transcript splicing, and promoter activityfollowing transfer to protoplasts by electroporation. PlantMolecular Biology, 18, 675-689.

[4]Cornejo M J, Luth D, Blankenship K M, Anderson O D, BlechlA E. 1993. Activity of a maize ubiquitin promoter in transgenicrice. Plant Molecular Biology, 23, 567-581.

[5]Della-Cioppa G, Bauer S C, Klein B K, Shah D M, Fraley R T,Kishore G M. 1986. Translocation of the precursor of 5-enolpyruvylshikimate 3-phosphate synthase intochloroplasts of higher plants in vitro. Procedings of theNational Academy of Science of the USA, 83, 6873-6877.

[6]Dill G M. 2005. Glyphosate-resistant crops: History, statusand future. Pest Management Science, 61, 219-224.

[7]Gruys K J, Marzabadi M R, Pansegrau P D, Sikorski J A. 1993.Stead-ystate kinetic evaluation of the reverse reaction forEscherichia coli 5-enolpyruvylshikimate-3-phosphatesynthase. Archives of Biochemistry and Biophysics, 304, 345-351.

[8]Herman K M, Weaver L M. 1999. The shikimate pathway.Annual Review of Plant Physiology and Plant MolecularBiology, 50, 473-503.

[9]Howe A R, Gasser C S, Brown S M, Padgette S R, Hart J, ParkerG B, Fromm M E, Armstrong C L. 2002. Glyphosate as aselective agent for the production of fertile transgenic maize(Zea mays L.). Molecular Breeding, 10, 153-164.

[10]Hu T, Metz S, Chay C, Zhou H P, Biest N, Chen G, Cheng M,Feng X, Radionenko M, Lu F, Fry J. 2003. Agrobacteriummediatedlarge-scale transformation of wheat (Triticumaestivum L.) using glyphosate selection. Plant Cell Reports,21, 1010-1019.

[11]Kishore G M, Padgette S R, Fraley R T. 1992. History ofherbicide-tolerant crops, methods of development and currentstate of the art-emphasis on glyphosate tolerance. WeedTechnology, 6, 626-634.

[12]Lin C Y, Fang J, Xu X L, Zhao T, Cheng J A, Tu J M, Ye G, ShenZ C. 2008. A built-in strategy for containment of transgenicplants: creation of selectively terminable transgenic rice. PLoSONE, 3, e1818.McElroy D, Blowers A, Jenes B, Wu R. 1991. Construction ofexpression vectors based on the rice actin 1 (Act1) 5 regionfor use in monocot transformation. Molecular Genetics andGenomics, 231, 150-160.

[13]McElroy D, Zhang W, Wu R. 1990. Isolation of an efficient actinpromoter for use in rice transformation. The Plant Cell, 2,163-171.

[14]Padgette S R, Re D B, Gasser C S, Eichholtz D A, Frazier R B,Hironaka C M, Levine E B, Sha D M, Fraley R T, Kishore GM. 1991. Site-directed mutagenesis of a conserved region ofthe 5-enolpyruvylshikimate-3-phosphate synthase activesite. The Journal of Biological Chemistry, 266, 22364-22369.

[15]Schledzewski K, Mendel R. 1994. Quantitative transient geneexpression: comparison of the promoters for maizepolyubiquitin 1 rice actin 1, maize derivedEmu and CaMV35S in cells of barley, maize and tobacco. TransgenicResearch, 3, 249-255.

[16]Schmid J, Amrhein N. 1995. Molecular organization of theshikimate pathway in higher plants. Phytochemistry, 39, 737-749.

[17]Xu X, Kawasaki S, Fujimura T, Wang C. 2005. A protocol forhigh-throughput extraction of DNA from rice leaves. PlantMolecular Biology Reporter, 23, 291-295.

[18]Zhou M, Xu H L, Wei X L, Ye Z Q, Wei L P, Gong W M, WangY Q, Zhu Z. 2006. Identification of a glyphosate-resistantmutant of rice 5-enolpyruvylshikimate-3-phosphatesynthase using a directed evolution strategy. PlantPhysiology, 140, 184-195.
[1] ZHANG Chun, YU Chao-jie, ZHANG Tai-jie, GUO Wen-lei, TIAN Xing-shan. Transcriptomic analysis reveals the transcription factors involved in regulating the expression of EPSPS gene, which confers glyphosate resistance of goosegrass (Eleusine indica)[J]. >Journal of Integrative Agriculture, 2021, 20(8): 2180-2194.
[2] SONG Ya-na, CHEN Zai-jie, WU Ming-ji, LI Gang, WANG Feng. Changes in bacterial community and abundance of functional genes in paddy soil with cry1Ab transgenic rice[J]. >Journal of Integrative Agriculture, 2021, 20(6): 1674-1686.
[3] XIAO Pei-ying, LIU Yi, CAO Yue-ping.
Overexpression of G10-EPSPS in soybean provides high glyphosate tolerance
[J]. >Journal of Integrative Agriculture, 2019, 18(8): 1851-1858.
[4] GUO Wen-fang, Kevin Yueju Wang, WANG Nan, LI Jun, LI Gang-qiang, LIU De-hu. Rapid and convenient transformation of cotton (Gossypium hirsutum L.) using in planta shoot apex via glyphosate selection[J]. >Journal of Integrative Agriculture, 2018, 17(10): 2196-2203.
[5] YAN Shu-feng, Sher Muhammad, LIU Hai-fang, TIE Shuang-gui, SUN Shu-ku. Identifying glyphosate-tolerant maize by soaking seeds in glyphosate solution[J]. >Journal of Integrative Agriculture, 2018, 17(10): 2302-2309.
[6] HAN Jiao, YU Guo-hong, WANG Li, LI Wei, HE Rui, WANG Bing, HUANG Sheng-cai, CHENG Xian-guo. A mitochondrial phosphate transporter, McPht gene, confers an acclimation regulation of the transgenic rice to phosphorus deficiency[J]. >Journal of Integrative Agriculture, 2018, 17(09): 1932-1945.
[7] RONG Rui-juan, WU Peng-cheng, LAN Jin-ping, WEI Han-fu, WEI Jian, CHEN Hao, SHI Jia-nan, HAO Yu-jie, LIU Li-juan, DOU Shi-juan, LI Li-yun, WU Lin, LIU Si-qi, YIN Chang-cheng, LIU Guo-zhen. Western blot detection of PMI protein in transgenic rice[J]. >Journal of Integrative Agriculture, 2016, 15(4): 726-734.
[8] LI Li, GUO Cheng, WANG Biao, ZHOU Tong, LEI Yang, DAI Yu-hua, HE Wen, LIANG Chun, WANG Xi-feng. RNAi-mediated transgenic rice resistance to Rice stripe virus[J]. >Journal of Integrative Agriculture, 2016, 15(11): 2539-2549.
[9] Janel L Huffman, Chance W Riggins, Lawrence E Steckel, Patrick J Tranel. The EPSPS Pro106Ser substitution solely accounts for glyphosate resistance in a goosegrass (Eleusine indica) population from Tennessee, United States[J]. >Journal of Integrative Agriculture, 2016, 15(06): 1304-1312.
[10] HUANG Yao, LI Ji-kun, QIANG Sheng, DAI Wei-min, SONG Xiao-ling. Transgenic restorer rice line T1c-19 with stacked cry1C*/bar genes has low weediness potential without selection pressure[J]. >Journal of Integrative Agriculture, 2016, 15(05): 1046-1058.
[11] ZHANG Chun, FENG Li, HE Ting-ting, YANG Cai-hong, CHEN Guo-qi, TIAN Xing-shan. Investigating the mechanisms of glyphosate resistance in goosegrass (Eleusine indica) population from South China[J]. >Journal of Integrative Agriculture, 2015, 14(5): 909-918.
[12] JIN Yong-mei, MA Rui, YU Zhi-jing, WANG Ling, JIANG Wen-zhu, LIN Xiu-feng. Development of lepidopteran pest-resistant transgenic japonica rice harboring a synthetic cry2A* gene[J]. >Journal of Integrative Agriculture, 2015, 14(3): 423-429.
[13] QIANG Xiao-jing, YU Guo-hong, JIANG Lin-lin, SUN Lin-lin, ZHANG Shu-hui, LI Wei, CHENG Xian-guo. Thellungiella halophila ThPIP1 gene enhances the tolerance of the transgenic rice to salt stress[J]. >Journal of Integrative Agriculture, 2015, 14(10): 1911-1922.
[14] JIANG Xian-bin, HUANG Qian, LING Yan, CHEN Yu-chong, XIAO Guo-ying, HUANG Suo-sheng, WU Bi-qiu, HUANG Feng-kuan, CAI Jian-he, LONG Li-ping. Functional and numerical responses of Cyrtorhinus lividipennis to eggs of Nilaparvata lugens are not affected by genetically modified herbicide-tolerant rice[J]. >Journal of Integrative Agriculture, 2015, 14(10): 2019-2026.
[15] LU Zeng-bin, HAN Nai-shun, TIAN Jun-ce, PENG Yu-fa, HU Cui, GUO Yu-yuan, SHEN Zhicheng, YE Gong-yin. Transgenic cry1Ab/vip3H+epsps Rice with Insect and Herbicide Resistance Acted No Adverse Impacts on the Population Growth of a Non-Target Herbivore, the White-Backed Planthopper, Under Laboratory and Field Conditions[J]. >Journal of Integrative Agriculture, 2014, 13(12): 2678-2689.
No Suggested Reading articles found!