華九頭獅子草（Dicliptera chinensis Juss.）為爵床科草本植物，分佈於台灣全省低海拔地區，此植物對於除草劑嘉磷塞(glyphosate)具高度耐性，經由此藥劑測定，華九頭獅子草之ED50值為紫花霍香薊、野甘草、野莧、鬼針等種雜草之2-7倍。耐藥特性為華九頭獅子草之原有特質，可遺傳於子代。噴施20 mM嘉磷塞造成華九頭獅子草葉片生長速率減慢，幼葉葉色淡化，葉肉柵狀組織及海綿組織，呈現不規則緊密排列，導管及篩管數目減少。但是施藥後7日，幼葉細胞之細胞核、葉綠體、粒線體及澱粉粒等皆完整。以14C-嘉磷塞測試，施藥後6小時藥劑即進入植體，施藥後168小時，根部累積量最大，嘉磷塞於華九頭獅子草及紫花霍香薊-嘉磷塞敏感植物之吸收與傳導率相似。施藥後14日，植體內無嘉磷塞代謝物-aminomethylphosphonic acid (APMA)之檢出，顯示華九頭獅子草之耐藥特性與藥劑代謝無顯著關聯性。20 mM 嘉磷塞處理後14日，華九頭獅子草幼葉總胺基酸含量因leucine及phenylalanine增加而增高，成熟葉之phenylalanine、serine、proline及methionine及根部phenylalanine含量略增加。華九頭獅子草之幼葉、成熟葉及根部shikimic acid含量於施藥後增加為未施藥者之5-9倍。幼葉及成熟葉二次代謝物benzoic acid、chlorogenic acid 及t-cinnamic acid含量增加1.5至3倍。幼葉及根部之protocatechuic acid 含量於施藥後14日增加為未施藥者之2.5及6.5倍。華九頭獅子草之5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)活性以幼葉最高。華九頭獅子草及紫花霍香薊幼葉噴施嘉磷塞，二者EPSPS之I50值分別為6.8 mM及0.8 mM。藥劑處理後24.5小時，華九頭獅子草幼葉仍具有活性，紫花霍香薊者已喪失活性。華九頭獅子草EPSPS具二個cDNAs：EPSPS-1及EPSPS-2 (1891 bp, open reading frame為1548 bp，transit peptide為210 bp)，二者差異為26個鹼基不同。華九頭獅子草EPSPS胺基酸與矮牽牛、菸草、番茄及阿拉伯芥者具84-88%相似度。此植物EPSPS mRNA之表現以幼葉最強，其次為花、根及莖部。嘉磷塞處理後8-24小時，華九頭獅子草EPSPS mRNA表現明顯增加，以16小時表現量最多。紫花霍香薊EPSPS mRNA於 8小時後即大量降低。華九頭獅子草 mature EPSPS蛋白質含量於2 mM嘉磷塞浸藥處理後8.5小時開始增加，至16.5小時增加為未施藥者之55倍。這些證據顯示華九頭獅子草對嘉磷塞之耐藥特性有二原因：一、華九頭獅子草具高活性之EPSPS。二、經由嘉磷塞處理後，可誘導華九頭獅子草EPSPS mRNA之表現並使EPSPS蛋白質含量增加。

Dicliptera chinensis Juss. is a naturally occurring glyphosate-tolerant plant. It is an indigenous Acanthaceae of Taiwan and East Asia. In the last two decades, this plant has become a dominant weed in some of the lowland orchards in central Taiwan as a result of chemical weed control. D. chinensis survived after spraying with 3.3 kg ha-1 of glyphosate. Greenhouse tests showed that ED50 for D. chinensis was 2-7 times higher than that for other common weeds species. This tolerance of D. chinensis to glyphosate was closely related with it’s developmental stages. The tolerance of D. chinensis to glyphosate was not derived from selection by repeated use of this herbicide. There was no glyphosate-sensitive D. chinensis variety in Taiwan. The absorption, translocation and degradation of glyphosate in D. chinensis were similar to Ageratum houstonianum, suggested that these were not the basis of the tolerance. The accumulation of shikimate in young leaf was increased 5- to 9-fold in leaf and root tissues. The levels of phenylalanine, tyrosine and tryptophan in young leaves, and phenylalanine in mature leaves were not decreased at 14 day after treatment. These changes were different from those of glyphosate-sensitive plants. Protocatechuate (PCA) levels were 2.5- and 6.5-fold higher than those of the control in young leaves and roots. The less accumulation of shikimate, PCA, gallic acid and increase of aromatic amino acids in herbicide-treated D. chinensis plants could explain that the shikimate pathway of this plant did not completely be blocked by glyphosate. In comparison to the glyphosate-sensitive weed A. houstonianum, D. chinensis had higher 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity even before glyphosate treatment. Glyphosate treatment resulted in significant increase of the EPSPS mRNA and protein level. This increase was apparent at about 8 h after glyphosate treatment, and peaked at about 16 h. The higher endogenous EPSPS activity and the induction by glyphosate could probably explain the tolerance to glyphosate. Two cDNAs representing two distinct EPSPS genes were isolated. Their sequences were highly homologous to the known plant and bacterial EPSPS. Northern and western blot analyses revealed differential tissue distribution. The analysis of mRNA and protein expression also suggested possible posttranscriptional regulation.

目 錄
中文摘要…………………………………………………………………1
英文摘要…………………………………………………………………3
縮寫列表…………………………………………………………………5
第一章 前言
一、 概說：華九頭獅子草與嘉磷塞………………………………..7
二、 嘉磷塞之作用機制……………………………………………..8
三、 嘉磷塞於植體之吸收、傳導與代謝…………………………10
四、 嘉磷塞對植物形態、胺基酸、shikimic acid…………….…..15
及二次代謝物之影響
五、 嘉磷塞之抗藥性及EPSPS 之特性…………………………..18
第二章 材料與方法
一、 華九頭獅子草生長與嘉磷塞之劑量反應關係………………..23
二、 嘉磷塞於華九頭獅子草之吸收傳導與代謝…………………..25
三、 嘉磷塞對華九頭獅子草葉片組織及胞器之影響……………. 30
四、 嘉磷塞對華九頭獅子草生理生化反應之影響………………. 36
五、 華九頭獅子草之EPSPS 基因與其蛋白質特性……………….44
第三章 結果
一、華九頭獅子草與嘉磷塞之劑量反應關係……………………….61
二、嘉磷塞於華九頭獅子草之吸收傳導與代謝…………………….63
三、嘉磷塞對華九頭獅子草形態、生理生化反應之影響………….64
四、華九頭獅子草之EPSPS 基因與其蛋白質特性…………………69
第四章 討論…………………………………………………………….73
圖列……………………………………………………………………...88
表列…………………………………………………………………….113
參考文獻……………………………………………………………….125
附錄…………………………………………………………………….146

參 考 文 獻
張韻如、蔡文福。2001。水稻及青江菜幼苗根部對嘉磷塞之吸收特性。雜草學會會刊。22(1): 13-29。
蔣慕琰、蔣永正、袁秋英、徐玲明。1995。雜草防除。臺灣農家要覽。農作篇(三)。行政院農委會。台北。317-334頁。
費雯綺、王玉美、張國輝、李朗照、廖莉莉 編。1999。植物保護手冊。臺灣省政府農林廳。南投。658-676頁。
鄭武燦。2000。臺灣植物圖鑑。國立編譯館。臺北。1320頁。
劉寧顏。1993。重修臺灣省通志。卷二 土地志、博物篇、植物篇第二冊。臺灣省文獻委員。南投。1048頁。
臺灣植物誌編輯委員會 編。1998。臺灣植物誌。臺灣植物誌編輯委員會。臺北。IV. 627-630。
正宗嚴敬 編。1936。最新臺灣植物總目錄。臺北市國大學理農學部植物分類生態學教室內KUDOA編輯部。臺北。197頁。
Amrhein, N. and Hollander, H. 1981. Light promotes the production of shikimic acid in buckwheat. Naturwissenschaften. 68: 3.
Amrhein, N., Deus, B., Gehrke, P. and Steinrücken, H. C. 1980. The site of the inhibition of the shikimate pathway by glyphosate. Plant Physiol. 65:830-834.
Anderson, K. S. and Johnson, K. A. 1990. Kinetic and structural analysis of enzyme intermediates: lessons from EPSP synthase. Chem. Rev. 90: 1131-1149.
Anderson, K. S., Sikorski, J. A. and Johnson, K. A. 1988. Evaluation of 5-enolpyruvylshikimate-3-phosphate synthase subatrate and inhibitor binding by stopped-flow and equilibrium fluorescence measurements. Biochemistry 27: 1604-1610.
Arnaud, L., Nurit, F., Ravanel, P. and Tissut, M. 1994. Distribution of glyphosate and of its target enzyme inside wheat plants. Pestic. Sci. 40: 217-223.
Arredodo, P. R., Moran, J. F., Sarath, G., Luan, P. and Klucas, R.V. 1997. Molecular cloning of the cowpea leghemoglobin II gene and expression of its cDNA in Escherichia coli purification and characterization of the recombinant protein. Plant Physiol. 114: 493-500.
Ashton, F. M. and Crafts, A. S. 1981. Mode of action of herbicide. 2nd edition. Wiley-Inter Sci. Publi. New York. pp. 236-253.
Barry, G., Kishore, G., Padgette, S., Taylor, M., Kolacz, K., Weldon, M., Re, D., Eichholtz, D., Fincher, K. and Halls, L. 1992. p. 139-145 in B. K. Singh, H. E. Flores, and J. C. Shannon, eds. Biosynthesis and Molecular Recognition of amino acid in plants. Am. Soc. Plant Physiol. Rockville, MD.
Becerril, J. M., Duke, S. O. and Lydon, J. 1989. Glyphosate effects on shikimate pathway products in leaves and flowers of velvetleaf. Phytochemistry. 28(3): 695-699.
Benfey, P. N., Takatsuji, H., Ren, L., Shah, D. M. and Chua, N. H. 1990. Sequence requirements of the 5-enolpyruvylshikimate-3-phosphate synthase 5’-upstream region for tissue-specific expression in flowers and seedlings. The Plant Cell 2: 849-856.
Bently, R. 1990. The shikimate pathway-A metabolic tree with many branches. P. 307-384. in G. D. Fasman, ed. Critical Reviews in Biochemistry and Molecular Biology, v. 25. CRC Press, Boca Raton, FL.
Berlin, J. and Widholm, L. 1981. Effects of glyphosate on shikimic acid accumulation in tobacci cell cultures with low and high yields of cinnamoyl putrescines. Zeitschrift fur Naturforschung. 36C, 210-214.
Boerboom, C. M., Ehlke, N. J., Wyse, D. L. and Somers, D. A. 1991. Recurrent selection for glyphosate tolerance in birdsfoot trefoil. Crop Sci. 31: 1124-1129.
Boerboom, C. M., Wyse, D. L. and Somers, D. A. 1990. Mechanism of glyphosate tolerance in birdsfoot trefoil (Lotus corniculatus). Weed Sci. 38:463-467.
Boocock, M. R. and Coggins, J. R. 1983. Kinetics of 5- enolpyruvylshikimate -3- phosphate synthase inhibition by glyphosate. FEBS Lett. 154: 127-133.
Bradshaw, L. D., Padgette, S. R., Kimball, S. L. and Wells, B. H. 1997. Perspectives on glyphosate resistance. Weed Technology 11: 189-198.
Brecke, B. J. 1976. Studies on the mechanism of action of N-(phosphonomethyl)-glycine (glyphosate)-Ph. D. Dissertation, Cornell University. Ithaca, NY.
Brecke, B. J. and Duke W. B. 1980. Effect of glyphosate on intact bean plants (Phaseolus vulgaris L.) and isolated cells. Plant Physiol. 66: 656-659.
Campbell, W. F., Evans, J. O. and Reed, S. C. 1976. Effects of glyphosate on chloroplast ultrastructure of quackgrass mesophyll cells. Weed Sci. 24(1): 22-25.
Canal, M. J., Albuerene, M., Tamesand, R. S. and Fernandez, B. 1990. Glyphosate injury on Cyperus esculantus leaves and bulbs: Histological study. Weed Res. 30: 117-122.
Cioppa, G. D., Bauer, S. C., Klein, B. K., Shah, D. M., Fraley, R. T. and Kishore, G. K. 1986. Translocation of the precursor of 5-enolpyruvylshikimate-3-phosphate synthase into chloroplast of higher plants in vitro. Proc. Natl. Acad. Sci. 83: 6873-6877.
Claus, J. S. and Behrens, R. 1976. Glyphosate translocation and quackgrass rhizome bud kill. Weed Sci. 24: 149-152.
Cole, D. J., Dodgemand, A. D. and Caselet, J. C. 1980. Some biochemical effects of glyphosate on plant meristems. J. exp. Bot. 31: 1665-1674.
Comai, L., Sen, L. and Stalker, D. M. 1983. An altered aroA gene product confers resistance to the herbicide glyphosate. Science 221: 370-371.
Cotterman, J. C. and Saari, L. L. 1992. Rapid metabolic inactivation is the basis for cross-resistance to chlorsulfuron in diclofop-methyl-resistant rigid ryegrass (Lolium rigidum L.) biotype sr4/84. Pestic. Biochem. Pyhsiol. 43: 182-192.
Coupland, D. 1985. Metabolism of glyphosate in plants, in “The Herbicide Glyphosate” (E. Grossbard and D. Atkinson, Eds.), pp. 25-47, Butter-worths, London.
deGennaro, F. P. and Weller, S. C. 1984. Differential susceptibility of field bindweed (Convolvulus arvensis) biotype to glyphosate. Weed Sci. 32: 472-476.
della-Cioppa, G., Bauer, S. C., Klein, B. K., Shah, D. M. Fraley, R. T. and Kishore, G. 1986. Translocation of the precusor of 5-enolpyruvylshikimate-3-phosphate synthase into chloroplasts of higher plants in vitro. Proc. Natl. Acad. Sci. USA. 83: 6873-6877.
della-Cioppa, G. and Kishore, G. M. 1988. Import of a precursor protein into chloroplast is inhibited by the herbicide glyphosate. The EMBO J. 7: 1299-1305.
Devine, M., Duke, S. O. and Fedtke, C. 1993. Inhibition of amino acid biosynthesis. In Physiology of herbicide action. pp. 252-263. P T R Prentice-Hall Press, Englewood Cliffs. New Jersey.
Duke, S. O. 1988. Glyphosate. In herbicides-chemistry, degradation and mode of action. P. C. Kearney and D. D. Kaufman, eds. New York: Dekker, pp. 1-70.
Duke, S. O., Christy, A. L., Hess, F. D. and Holt, J. S. 1991. Herbicide-resistant crops. Comments from Counc. Agric. Sci. Technol. P. 1-28.
Duke, S. O. and Hoagland, R. E. 1978. Effects of glyphosate on metabolism of phenolic compounds. I. Induction of phenylalanine ammonia-lyase activity in dark-grown maize roots. Plant Sci. Lett. 11: 185-190.
Duke, S. O., Hoagland, R. E. and Elmore, C. D. 1979. Effects of glyphosate on metabolism of phenolic compounds. IV. Phenylalanine ammonia-lyase activity, free amino acids, and soluble hydroxyphenolic compounds in axes of light-grown soybeans. Physiol. Plant. 46: 307-317.
Duke, S. O. 1996. Herbicide-resistant crops: Agricultrural, environmental, economic, regulatory, and technical aspects. CRC Press, Inc., USA pp. 53-84.
Duncan, K., Lewendon, A. and Coggins, J. R. 1984. The complete amino acid sequence of Escherichia coli 5-enolpyruvylshikimate-3-phosphate synthase. FEBS Lett. 170: 59-63.
Duncan, C. N. and Weller, S. C. 1987. Heritability of glyphosate susceptibility among biotype of field bindweed. J. Heed. 78: 257-260.
Duncan, K., Lewedon, A. and Coggins, J. R. 1984. The purification of 5-enolpyruvylshikimate-3-phosphate synthase from an overproducing strain of Escherichia coli. FEBS 165: 121-127.
Dyer, W. E. 1994. Resistance to glyphosate. In Herbicide Resistance in Plants: Biology and Biochemistry, edited by Powles, S. B. and Holtum, J. A. M. Boca-Raton: Lewis, pp. 229-241.
Dyer, W. E., Wellwe, S. C., Bressan, R. A. and Herrmann, K. M. 1988. Glyphosate tolerance in tobacco (Nicotiana tabacum L.). Plant Physiol. 88: 661-666.
Editorial committee of the Flora of Taiwan. 1978. Flora of Taiwan. IV. Angiospermae, Taipei, Taiwan, ROC pp. 627-630.
Eichholtz, D. A., Gasser, C. S. and Kishore, G. M. 1999. Glyphosate-tolerant 5-enolpyruvyl-3-phosphoshikimate synthases. US Patent 5: 866-775.
EI Ibaoui, H., Delrot, H., Besson, J. and Bonnemain. J. L. 1986. Uptale and release of a phloem-mobile (glyphosate) and of a non phloem-mobile (iprodione) xenobiotic by broadbean leaf tissues. Physiol. Veg. 24: 431-442.
Evans, F. J. and Schmidt, R. J. 1976. Two new toxins from the latex of
Euphorbia poisonii. Phytochemistry 15:333-335
Fernandez, C. H. and Bayer, D. E. 1977. Penetration, translocation and toxicity of glyphosate in bermudagrass (Cynodon dactylon L.). Weed Sci. 25: 396-400.
Forlani, G., Nielsen, E. and Racchi, M. L. 1992. A glyphosate-resistant 5-enol-pyruvyl-shikimate-3-phosphate synthase confers tolerance to a maize cell line. Plant Sci. 85: 9-15.
Forlani, G., Parisi, B. and Nielsen, E. 1994. 5- enol- pyruvyl- shikimate -3- phosphate synthase from Zea mays cultured cells. Plant physiol. 105: 1107-1114.
Franz, J. E., Mao, J. K. and Sikorski, J. A. 1993. Glyphosate’s molecular mode of action: A review of glyphosate and EPSPS biochemistry. Am. Chem. Soc. Monogr. 13: 1-87.
Gasser, C. S., Winter, J. A., Hironaka, C. K. and Shah, D. M. 1988. Structure, expression, and evolution of the 5-enolpyruvylshikimate-3-phosphate synthase genes of petunia and tomato. J. Biol. Chem. 263: 4280-4289.
Giesy, J. P., Dodson, S. and Solomon, K. R. 2000. Ecotoxocological risk assessment for Roundup herbicide. Reviews of Environmental Contamination and Toxicology 167: 5-120.
Goldsbrough, P. B., Hatch, E. M., Huang, B., Kosinki, W. G., Dyer, W. E., Hermann, K. M. and Weller, S. C. 1990. Gene amplification in glyphosate tolerant tobacco cells. Plant Sci. 72: 53-62.
Gottrup, O., O’sullivan, P. A., Schraa, R. J. and Vanden, W. H. 1976. Uptake, translocation, metabolism and selectivity of glyphosate in Canada thistle and leafy spurge. Weed Res. 16: 197-201.
Gougler, J. A. and Geiger, D. R. 1981. Uptake and distribution of N-(phosphonomethyl) glycine in sugar beet plants. Plant Physiol. 68: 668-672.
Gressel, J. 1996. Fewer constrains than proclaimed to the evolution of glyphosate-resistant weeds. Resistant Pest Management 8: 2-5.
Gresshoff, P. M. 1979. Growth inhibition by glyphosate and reseveral of its action by phenylalanine and tyrosine. Aust. J. Plant Physiol. 6: 177-185.
Gronwald, J. W., Anderson, R. N. and Yee, C. 1989. Atrazine resistance in velvetleaf (Abutilon threophrasti) due to enhanced atrazine detoxification. Pestic. Biochem. Physiol. 34: 149-163.
Grossbard, E. and Atkinson, D. 1985. The herbicide glyphosate. Butterworth and Company. Ltd., London. P. 490.
Gruys, K. J., Walker, M. C. and Sikorski, J. A. 1992. Substrate synergism and the steady-state kinetic reaction mechanism for EPSP synthase from E. coli. Biochemistry 31: 5534-5544.
Gruys, K. J., Marzabadi, M. R., Pansegrau, P. D. and Sikorski, J. A. 1993. Steady-state kinetic evaluation of the reverse reaction for Escherichia coli 5-enolpyruvylshikimate-3-phosphate synthase. Arch. Biochem. Biophys. 304: 345-451.
Haderlie, L. C., Widholm, J. M. and Slife, F. W. 1977. Effect of glyphosate on carrot and tobacco cells. Plant Physiol. 60: 40-43.
Harring, T., Streibig, J. C. and Husted, S. 1998. Accumulation of shikimic acid: a technique for screening glyphosate efficacy. J. Agric. Food Chem. 46:3306-4412.
Haslam, E. 1974. The shikimate pathwany. Wiley. New York.
Herrmann, K. M. and Weaver, L. M. 1999. The shikimate pathway. Annu. Rev. Plant Physiol. Plant Mol. Biol. 50: 473-503.
Hetherington, P. R., Marshall, G., Kirkwood, R. C. and Warner, J. M. 1998. Absorption and efflux of glyphosate by cell suspensions. J. Exp. Bot. 49: 527-533.
Hoagland, R. E., Duke, S. O. and Elmore, C. D. 1978. Effects of glyphosate on metabolism of phenolic conpounds.ⅡInfluence on soluble hydroxyphenolic compound, free amino acid and soluble protein levels in dark-grown maize roots. Plant Sci. Lett. 13: 291-299.
Hoagland, R. E., Duke, S. O. and Elmore, C. D. 1979. Effects of glyphosate on metabolism of phenolic compounds. III. Phenylalanine ammonia-lyase activity, free amino acid, and soluble hydroxyphenolic compounds in axes of light-grown soybean. Physiol. Plant. 46: 357-366.
Hollander-Czytko, H. and Amrhein, N. 1987. 5-enolpyruvylshikimate-3-phosphate synthase, the target enzyme of the herbicide glyphosate, is synthasized as a precursor in a higher plant. Plant Physiol. 83: 229-231.
Hollander-Czytko, H., Johanning, D., Mayer, H. E. and Amrhein, N. 1988. Molecular basis for the overproduction of 5-enolpyruvylshikimate-3-phosphate synthase in a glyphosate-tolerant cell suspension culture of Corydalis sempervirens Plant Mol. Biol. 11: 215-220.
Hollander-Czytko, H., Sommer, I. And Amrhein, N. 1992. Glyphosate tolerance of cultured Corydalis sempervirens cells is acquired by an increased rate of transcription of 5-enolpyruvylshikimate 3-phosphate synthase as well as by a reduced turnover of the enzyme. Plant Mol. Biol. 20: 1029-1036.
Holt, J. S. 1992. History of identification of herbicide-resistant weeds. Weed Technol. 6: 615-620.
Holt, J. S. and Lebaron, H. M. 1990. Significance and distrubution of herbicide resistance. Weed Technol. 4: 141-149.
Horsch, R. B., Fraley, R. T.,Rogers, S. G., Klee, H. J., Fry, J., Hinchee, M. A. W. and Shah, D. S. 1988. Agrobacterium-mediated gene transfer to plants: engineering tolerance to glyphosate. Iowa State J. Res. 62: 487-502.
http://www.weedscience.com 2002
Huynh, Q. K. 1988. Evidence for a reactive γ-carboxyl group (Glu-418) at the herbicide glyphosate binding site of 5-enolpyrulshikimate-3-phosphate synthase from Escherichia coli. J. Biol. Chem. 263: 11631-11635.
Huynh, Q. K., Bauer, S. C., Bild, G. S., Kishore, G. M. and Borgmenyer, J. R. 1988. Site-directed mutagenesis of petunia hybrida 5- enolpyruvyl shikimate-3-phosphate synthase: Lys-23 is essential for substrate binding. J Biol. Chem. 24: 11636-11639.
Jacob, G. S., Garbow, J. R., Hallas, L. E., Kimack, N. M., Kishore, G. M. and Schaefer, J. 1988. Metabolism of glyphosate in Pseudomonas sp. strain LBr, Appl. Environ. Microbiol. 54(12): 2953-2958.
Jaworski, E. G. 1972. Mode of action of N-phosphonomethyl-glyine: inhibition of aromatic amino acid biosynthesis. J. Agric. Food Chem. 20: 1195-1198.
Jensen, R. A. 1985. The shikimate / arogenate pathway: link between carbohydrate metabolism and secondary metabolism. Physiol. Plant 66: 164-168.
Johnson, D. T., Van Wijk, A. J. P. and Kilpatrick, D. 1989. Selection for tolerance to gltphosate in fine-leaved Festuca species. Proc. 6th Int. Turgrass Res. Conf. P. 103-105.
Johnson, D. T. and Faulkner, J. S. 1991. Herbicide resistance in the Graminaceae-A plant breeder’s view. P. 319-330. in J. C. Caseley, G. W. Cussans and Atkin, R. K. eds. Herbicide resistant weeds and crops. Butterworth-Heinemann Ltd., Oxford, U. K.
Khurana, J. P., Maheshwari, S. C. 1986. A comparison of the effects of chelates, salicylic acid and benzoic acid on growth and flowering of Spirodela polyrrhiza. Plant and Cell Physiology 27(5): 919-924.
Kim, T. W. and Amrhein, N. 1995. Glyphosate toxicity: I. Long term analysis of shikimic acid accumulation and chlorophyll degradation in tomato plant. Korean Jounal of Weed Scid. 15: 141-147.
Kishore, G. M., Brundage, L., Kolk, K., Padgette, S. R., Rochester, D., Huynh, K. and and della-Cioppa, G. 1986. Isolation, purification and characterization of a glyphosate tolerant mutant E. coli EPSP synthase. Fed. Rroc. 45: 1506.
Kishore, G. M. and Shah, D. M. 1988. Amino acid biosynthesis inhibitors as herbicides. Annu. Rev. Biochem. 57: 627-663.
Klevorn, T. B. and Wyse, D. L. 1984. Effect of leaf girdling and rhizome girdling on glyphosate and photoassimilate distribution in quackgrass (Agropyron repens). Weed Sci. 32: 402-407.
Komoba, D., Gennity, I. And Sandermann, H. 1992. Plant metabolism of herbicides with C-P bonds: glyphosate. Pestic. Biochem. And Physiol. 43: 85-94.
Koshiba, T. 1979. Organization of enzymes in the shikimate pathway in Phaseolus mungo seedlings. Plant Cell Physiol. 20: 677-670.
Lewedon, A. and Coggins, J. R. 1983. Purification of 5- enol- pyruvyl- shikimate-3-phosphate synthase from Escherichia coli. Biochem. J. 213: 187-191.
Lin, X., Kaul, S., Rounsley, S., Shea, T. P., Benito, M., Town, C. D., Fujii, C. Y., Mason, T., Bowman, C. L., Barnstead, M., Feldblyum, T. V., Buell, R., Ketchum, K. A., Lee, J., Ronning, C. M., Koo, H. L., Moffat, K. S., Cronin, L. A., Shen, M., Pai, G., Aken, S. V., Umayam, L., Tallon, L. J., Tallon, J., Gill, E., Adams, M. D., Carrera, A. J., Creasy, T. H., Goodman, H. M., Someville, C. R., Copenhaver, G. P., Preuss, D., Nierman, W. C., While, O., Eisen, J. A., Salzberg, L., Fraser, M. and Venter, C. 1999. Sequence and analysis of chromosome 2 of the plant Arabidopsis thaliana. Nature, 402: 761-768.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Kandall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265.
Lumsden, J. and Coggins, J. R. 1977. The subunit structure of the arom multienzyme complex of Neurospora crassa. Biochem. J. 161: 599-607.
Lydon, J. and Duke, S. O. 1988. Glyphosate induction of elevated levels of hydroxybenzoic acids in higher plants. J. Agric. Food Chem. 36: 813-818.
Lydon, J. and Duke, S. O. 1989. Pesticide effects on secondary metabolism of higher plants. Pestic. Sci. 25: 361-373.
Majumder, K., Selvapandiyan, A. and Fattam, F. A. 1995. 5- enol- pyruvyl- shikimate-3-phosphate synthase of Bacillus subtileu is an allosteric enzyme. Analysis of Arg 24, Pro 105, His 385 suggest a hidden phosphoenolpyruvate-binding site. Eur. J. Biochem. 229: 99-106.
Malik, J., Barry, G. and Kishore, G. 1989. The herbicide glyphosate. BioFactors 2(1): 17-25.
Marshall, G., Kirkwood, R. C. and Martin, D. J. 1987. Studies on the mode of action of asulam, aminotriazole and glyphosate in Equisetum arvense L. (field horsetail). II. The metabolism of [14C]asulam, [14C]aminotriazole and [14C]glyphosate, Pestic. Sci. 18: 65-77.
McCloskey, W. B. 1991. The absorption of atrazine, 2,4-D and glyphosate by wheat leaf protoplasts. pp. 148. Ph. D. Diss. Univ. of California, Davis.
Moorman, T. B., Becerril, J., M., Lydon, J. and Duke, S. O. 1992. Production of hydroxybenzoic acids by Bradyrhizobium japonicum strains after treatment with glyphosate. J. Agric. Food Chem. 40(2): 289-293.
Moss, S. R. and Rubin, B. 1993. Herbicide-resistant weeds: a worldwide perspective. J. Agric. Sci. 120: 141-148.
Mousdale, D. M. and Coggins, J. R. 1984. Purification and properties of 5-enolpyruvyl-shikimate-3-phosphate synthase from seedlings of Pisum sativum L. Planta 160: 78-83.
Mousale, D. M. and Coggins, J. R. 1985. Subcellular localization of the common shikimate-pathway enzymes in Pisum sativum L. Planta 163: 241-249.
Mullis, G. T. and Erlich, H. A. 1988. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: 487-491.
Nafziger, E. M., Wildholm, J. M., Steinrücken, H. C. and Killmer, J. L.1984. Selection and characterization of a carrot cell line tolerance to glyphosate. Plant Physiol. 76: 571-574.
Nilsson, G. 1977. Effects of glyphosate on the amino acid content in spring wheat plant. Swe. J. Agr. Res. 7: 153-157.
Nomura, N. S. and Hilton, H. W. 1977. The adsorption and degradation of glyphosate in five Hawaiian sugarcane soils. Weed Res. 17: 113-121.
Padgette, S. R., Re, D. B., Barry, G. F., Eichholtz, D. E., Delanny, X., Fuchs, R. L., Kishore, G. M. and Fraley, R. T. 1996. New weed control opportunities: development of soybeans with a Roundup Ready gene. In Herbicide-Resistant Crops: Agricultural, Environmental, Economic, Regulatory and Technical Aspects, edited by S. O. Duck. Boca Raton FL: CRC Press, pp 53-84.
Padgette, S. R., Re, D. B., Gassers, C. S., Eichholtz, D. A., Fraziers, R. B., Hironaka, C. M., Levine, E. B., Shah, D. M., Fraley, R. T. and Kishore, G. M. 1991. Site-directed mutagenesis of a conserved region of the 5-enolpyruvyl shikimate-3-phosphate synthase active site. J. Biol. Chem. 266: 22364-22369.
Padgette, S. R., Smith, C. E., Khaihuynh, Q. and Kishore, G. M. 1988. Arginine chemical modification of petunia hybrida 5- enol-pyruvyl shikimate synthase. Arch. Biochem. Biophy. 266: 254- 262.
Patel, V. B., Giles, N. H. 1979. Purification of the arom multienzyme aggregate fron Euglena gracilis. Biochem. Biophys. Acta. 567: 24-34.
Pipke, R. and Amrhein, N. 1988. Degradation of the phosphonate herbicide glyphosate by Arthrobacter atrocyaneus ATCC 13752. Apple. Environ. Microbiol. 54: 1293-1296.
Powles, S. B. Lorraine-Colwill, D. F., Dellow, J. J. and Preston, C. 1998. Evolved resistance to glyphosate in rigid ryegrass (Lolium rigidum) in Australia. Weed Sci. 46: 604-607.
Pratley, J., Baines, B., Eberbach, P., Incerti, M. and Broster, J. 1996. Glyphosate resistance in annual ryegrass. Proc. Eleventh Ann. Conf. Grassld. Soc. NSW. P. 122.
Putnam, A. R. 1976. Fate of glyphosate in deciduous fruit frees. Weed Sci. 24: 425-430.
Re, D. B., Padgette, S. R., Delannay, X., Kolacz, K. H., Nida, D. L., Peschke, V. M., Derting, C. W., Rogers, S. G., Edwards, J. W., Barry, G. F. and Biest, N. A. 1993. Petition of determination of nonregulated status: Soybeans with a Roundup Ready gene. Submitted to the United States Department of Agriculture, September, 1993.
Ream, J. E., Steinrücken, H. C., Porter, C. A. and Sikorski, J. A. 1988. Purification and properties of 5-enolpyruvylshikimate-3-phosphate synthase from dark-grown seedlings of Sorghum bicolor. Plant Physiol. 87: 232-238.
Reinbothe, S., Nelles, A. and Parthier, B. 1991. (N-phosphonometyl) glycine (glyphosate) tolerance in Euglena gracillis acquired by either overproduced or resistant 5-enolpyruvylshikimate-3-phosphate synthase. Eur. J. Biochem. 198: 365-373.
Rueppel, M. L., Brightwell, B. B., Schaefer, J. and Marvel, J. T. 1977. Metabolism and degradation of glyphosate in soil and water. J. Agric. Food Chem. 25: 517-528.
Schönbrunn, E., Eschenburg, S., Shuttleworth, W. A., Schloss, J. V., Amrhein, N., Evans, J. N.S. and Kabsch, W. 2001. Interaction of the herbicide glyphosate with its target enzyme 5-enolpyruvylshikimate 3-phosphate synthase in atomic detail. PNAS, 98: 1376-1380.
Schulz, A., Wengenmayer, F. and Goodman, H. M. 1990. Genetic engineering of herbicide resistance in higher plants. Crit. Rev. Plant Sci. 9(1): 1-15.
Sellin, C., Forlani, G., Dubois, J., Nielsen, E. and Vasseur, J. 1992. Glyphosate tolerance in Cichorium intybus L. var. Magdebourg. Plant Sci. 85: 223-231.
Shah, D. M., Horsch, R. B., Klee, H. J., Kishore, G. M., Winter, J. A., Tumer, N. E., Hironaka, C. M., Sanders, P. R., Gasser, C. S., Aykent, S. A., Siegel, N. R., Roger, S. G. and Fraley, R. T. 1986. Engineering herbicide tolerance in transgenic plants. Science 233: 478-481.
Shaner, D. L. and Lyon, J. L. 1980. Interaction of with aromatic amino acid on transpiration in Phaseolus vulgaris L. Weed Sci. 28: 31-35.
Shyr, Y. J., Hepburn, A. G. and Wildholm. J. M. 1992. Glyphosate selected amplification of the 5-enolpyruvylshikimate-3-phosphate synthase gene in cultured carrot cells. Mol. Gen. Genet. 232: 377-382.
Singer, S. R. and McDaniel, C. N. 1985. Selection of glyphosate-tolerant tobacco calli and the expression of this tolerance in regenerated plants. Plant Physiol. 78: 411-416.
Smith, C. M., Pratt, D. and Thompson, G. A. 1986. Increased 5-enolpyruvylshikimic acid 3-phosphate synthase activity in a glyphosate-tolerant variant strain of tomato cells. Plant Cell Rep. 5: 298-301.
Southern, E. 1975. Detection of special DNA sequences among fragments separated by gel electrophoresis. J Mol. Biol. 98: 503-517. Sprankle, P., Meggitt, W. F. and Penner, D. 1975. Absorption, action and translocation of glyphosate. Weed Sci. 23: 235-240.
Stalker, D. M., Hiatt, W. R. and Comai, L. 1985. A single amino acid substitution in the enzyme 5-enolpyruvylshikimate-3-phosphate synthase confers resistance to the herbicide glyphosate. J. Biol. Chem. 260: 4724-4728.
Stasiak, M. A., Hofstra, G., Payne, N. J., Prasad, R. and Fletcher, R. A. 1991. Alterations of growth and shikimic acid levels by sublethal glyphosate applications on pin cherry and trembling aspen. Can. J. For. Res. 21(7): 1086-1090.
Steinrücken, H. C. and Amrhein, N. 1980. The herbicide glyphosate is a potent inhibitor of 5-enolpyruvylshikimate-3-phosphate synthase. Biochem. Biophys. Res. Commun. 94: 1207-1212.
Steinrcüken, H. C. and Amrhein, N. 1984. 5-Enolpyuvylshikimate-3-phosphate synthase of Klebsiella pneumoniae. 2. Inhibition by glyphosate (N-phosphonometyl) glycine). Eur. J. Biochem. 143: 351-357.
Steinrücken, H. C., Schulz, A., Amrhein, N., Porter, C. A. and Fraley, R. 1986. Overproduction of 5-enolpyruvylshikimate-3-phosphate synthase in a glyphosate-tolerant Petunia hybrida cell line. Arch. Biochem. Biophys. 224: 169-178.
Theologis, A., Ecker, J. R., Palm, C. J., Federspiel, N. A., Kaul, F. S., White, O., Alonso, J., Altafi, H., Araujo, R., Bowman, C. L., Brooks, S. Y., Aprilchan, E. B., Chao, Q., Chen, H., Cheuk, R. F., Chin, C., Chung, M. K., Conn, L., Conway, A. B., Conway, A. R., Creasy, T. H., Dewar, K., Dunn, P., Etgu, P. E., Feldblyum, T. V., Feng, J., Fong, B., Fujii, C. Y., Gill, J. E., Goldsmith, A. D., Haas, B., Hansen, N. F., Hughes, B., Huizar, L., Hunter, J. L., Jenkins, J., Johnson-hopson, C., Khan, S., Khaykin, E., Kim, C. J., Koo, H. L., Kremenetskaia, I., Kurtz, D. B., Kwan, A., Lam, B., Langin-hooper, S., Lee, A., Lee, J. M., Lenz, C. A., Li, J. H., Li, Y., Lin, X., Liu, S. X., Liu, Z. A., Luros, J. S., Maiti, R., Marziali, A., Militscher, J., Miranda, M., Nguyen, M., Ninerman, W. C., Osborne, B. I., Pai, G., Peterson, J., Pham, P. K., Rizzo, M., Rooney, T., Rowley, D., Sakano, H., Salzberg, S. L., Schwartz, J. R., Shinn, P., Southwick, A. M., Sun, H., Tallon, L. J., Tambunga, G., Toriumi, M. J., Town, C. D., Utterback, T., Aken, S. V., Vaysberg, M., Vysotskaia, V. S., Walker, M. I., Wu, D., Yu, G., Fraser, C. M., Venter, J. C. and Davis, R. W. 2000. Sequence and analysis of chromosome 1 of the Arobidopsis thaliana. Nature 408: 816-820.
Tran, M., Baerson, S., Brinker, R., Casagrande, L., Faletti, M., Feng, Y., Nemeth, M., Reynolds, T., Rodriguez, D., Schafer, D., Stalker, D., Taylor, N., Teng, Y. and Dill, G. 1999. Characterization of glyphosate resistant Eleusine indica biotypes from Malaysia. pp. 527-536. Proceedings 17th Asia-Pacific Weed Science Society Conference. Thailand.
Tymonko, J. M. and Foy, C. L. 1978. Inhibition of protein synthesis by glyphosate. Plant Physiol. Ann. Suppl. 61: 41-45.
Tymonko, J. M. and Foy, C. L. 1978a. Influence of glyphosate on the metabolism of separated soybean leaf cells. Abstracts of the Weed Science Society of America. pp. 70-71.
Uotila, M., Evjen, K. and Iversen, T. H. 1980. The effects of glyphosate on the development and cell ultrastructure of white mustard (Sinapis alba L.) seedlings. Weed Res. 20: 153-158.
Vaughn, K. C. and Duke, S. O. 1986. Ultrastructural effects of glyphosate on Glycine max seedlings. Pest. Biochem. and Physiol. 26：56-65.Uotila, M., Gullner, G. and Kömives, T. 1995. Introducation of glutathione S-transferase activity and glutathione level in plants exposed to glyphosate. Physiol. Plant. 93: 689-694.
Wang, Y., Jones, J. D., Weller, S. C. and Goldsbrough, P. B. 1991. Expession and stability of amplified genes encoding 5-enolpyruvylshikimate-3-phosphate synthase in glyphosate-tolerant tobacco cells. Plant Mol. Biol. 17: 1127-1138.
Wells, B. H. and Appleby, A. P. 1992. Lactofen increase glyphosate-stimulated shikimate production in little mallow (Malva parviflora). Weed Sci. 40(2): 171-173.
Westwood, J. H. and Weller, S. C. 1997. Cellular mechanisms influence differential glyphosate sensitivity in field bindweed (Convolvulus arvensis). Weed Sci. 45: 2-11.
Williams, G. M., Kroes, R. and Munro, I. C. 2000. Safety evaluation and risk assessment of the herbicide Roundup and its active ingredient, glyphosate, for humans. Regulatory Toxicology and Pharmacology 31: 117-165.
Wyrill, J. B. and Burnside, O. C. 1976. Absorption, translocation and metabolism of 2,4-D and glyphosate in Common Milkweed and Hemp Dogbane. Weed Sci. 24(6): 557-566.
Ye, B. and Gressel, J. 2000. Trasient, oxidant-induced antioxidant transcript and enzyme levels correlate with greater oxidant-resistance in paraquat-resistant Conyza bonariensis. Planta 211: 30-61.
Yuan, C. I., Chen, Y. M. and Chiang, M. Y. 2001. Responses of Dicliptera chinensis to glyphosate. Plant Prot. Bull. Taiwan, 43: 29-38.