Alleviation of chlorimuron-ethyl toxicity to soybean by branched-chain amino acids or naphthalic anhydride

  • Mamdouh M. Nemat AllaEmail author
  • Nemat M. Hassan


Soybean seeds, soaked in branched-chain amino acids [Valine, Isoleucine and Leucine (VIL)] mixture or dressed in naphthalic anhydride (NA), were germinated for 10 days and treated with chlorimuron-ethyl for 120 h. Chlorimuron-ethyl significantly inhibited acetolactate synthase (ALS) activity in shoots of all samples during the first 72 h. Moreover, it increased Km but decreased Vmax, Kcat, Kcat/Km and Vmax/Km as well as the endogenous levels of valine, isoleucine and leucine particularly during the first 72 h. In concomitance, there were significant decreases in protein content and activities of nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT) as well as growth parameters during the first 72 h. However, VIL or NA mitigated the herbicide effect. So, the drop in Vmax, Kcat and Kcat/Km by chlorimuron-ethyl declares decreases in enzyme concentration, catalytic rate and catalytic efficiency, respectively. Whereas the increase in Km could indicate that chlorimuron-ethyl induced an inhibition for ALS of the mixed-type which would affect synthesis and structural integrity of the enzyme. This inhibition would diminish the formation of branched-chain amino acids and protein, particularly with the inhibition of N incorporation enzyme (NR, NiR, GS and GOGAT). Consequently, retardation and disturbances in formation of structural and functional protein would arise. Nonetheless, VIL mixture overcame chlorimuron-ethyl toxicity via compensating the drop in the endogenous branched-chain amino acids while NA might cause mitigation by detoxifying the herbicide.


Acetolactate synthase Branched-chain amino acids Chlorimuron-ethyl Naphthalic anhydride Soybean 



  1. Badran EG, Abogadallah GM, Nada RM, Nemat Alla MM (2015) Role of glycine in improving the ionic and ROS homeostasis during NaCl stress in wheat. Protoplasma 252:835–844CrossRefGoogle Scholar
  2. Bidlingmeyer BA, Cohen SA, Tarvin TL, Frost B (1987) A new, rapid, high sensitivity analysis of amino acids in food type samples. J Assoc Off Anal Chem 70:241Google Scholar
  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248CrossRefGoogle Scholar
  4. Deng F, Hatzios KK (2003) Characterization of cytochrome P450-mediated bensulfuron-methyl O-demethylation in rice. Pestic Biochem Physiol 74:102–104CrossRefGoogle Scholar
  5. Farver O, Kroneck PMH, Pecht I, Zumft WG (2006) Allosteric control of internal electron transfer in cytochromecd 1 nitrite reductase. Rend Fis Acc Lincei 17:213–220CrossRefGoogle Scholar
  6. Gioia M, Tomao L, Sbardella D, Ciaccio C, Tundo GR, Di Masi A, Fasciglione GF, Marini S, Cozza P, Ascenzi P, Coletta M (2017) Enzyme catalysis: the case of the prostate-specific antigen. Rend Fis Acc Lincei 28:229–237CrossRefGoogle Scholar
  7. Harborne JB (1984) Macromolecules. In: Harborne JB (ed) Phytochemical methods. Chapman and Hall, London, p 243CrossRefGoogle Scholar
  8. Hassan NM, Serag MS, El-Feky FM, Nemat Alla MM (2008) In vitro selection of mung bean and tomato for improving tolerance to NaCl. Ann Appl Biol 152:319–330CrossRefGoogle Scholar
  9. Hatzios KK (2000) Herbicide safeners and synergists. In: Roberts T (ed) Metabolism of agrochemicals in plants. Wiley, Chichester, pp 259–294Google Scholar
  10. Hecht U, Oelmüller R, Schmidt S, Mohr H (1988) Action of light, nitrate and ammonium on the levels of NADH- and ferridoxin- dependent glutamate synthases in the cotyledons of mustard seedlings. Planta 175:130–138CrossRefGoogle Scholar
  11. Huang Z, Sui B, Zhang C, Huang H (2019) The basis of resistance mechanism to mesosulfuron-methyl in Tausch’s goatgrass (Aegilops tauschii Coss.). Pestic Biochem Physiol 155:126–131CrossRefGoogle Scholar
  12. Kong X, Luo Z, Dong H, Li W, Chen Y (2017) Non-uniform salinity in the root zone alleviates salt damage by increasing sodium, water and nutrient transport genes expression in cotton. Sci Rep 7:2879CrossRefGoogle Scholar
  13. Lea PJ, Robinson SA, Stewart GR (1990) The enzymology and metabolism of glutamine, glutamate and asparagines. In: Miflin BJ, Lea PJ (eds) The biochemistry of plants, vol 16. Academic Press, NewYork, pp 121–159Google Scholar
  14. Long W, Malone J, Boutsalis P, Preston C (2019) Diversity and extent of mutations endowing resistance to the acetolactate synthase (AHAS)-inhibiting herbicides in Indian hedge mustard (Sisymbrium orientale) populations in Australia. Pestic Biochem Physiol In PressGoogle Scholar
  15. Marczewska K, Sadowski J, Rola H (2006) Changes in branched chain amino acids content in leaves of Apera spica-venti biotypes resistant and susceptible to chlorsulfuron. J Plant Prot Res 46:191–198Google Scholar
  16. Marquez AJ, Avile C, Forde BG, Wallsgrove RM (1988) Ferredoxin-glutamate synthase from barley leaves: rapid purification and partial characterization. Plant Physio Biochem 26:645–651Google Scholar
  17. Nagaoka S, Hirasawa M, Fukushima K, Tamura G (1984) Methyl viologen-linked nitrite reductase from bean roots. Agric Biol Chem 48:1179–1188Google Scholar
  18. Nakagawa H, Yonimura Y, Yamamota H, Sato T, Ogura N, Sato R (1985) Spinach nitrate reductase: purification, molecular weigh, and subunit composition. Plant Physiol 77:124–128CrossRefGoogle Scholar
  19. Nemat Alla MM, Hassan NM (1998) Efficacy of exogenous GA3 and herbicide safeners in protection of Zea mays from metolachlor toxicity. Plant Physiol Biochem 36:809–815CrossRefGoogle Scholar
  20. Nemat Alla MM, Hassan NM (2008) Recognition, implication and management of plant resistance to herbicides. Am J Plant Physiol 3:50–66CrossRefGoogle Scholar
  21. Nemat Alla MM, Hassan NM, El-Bastawisy ZM (2007) Differential influence of herbicide treatments on activity and kinetic parameters of C4 photosynthetic enzymes from Zea mays. Pestic Biochem Physiol 89:198–205CrossRefGoogle Scholar
  22. Nemat Alla MM, Badawi AM, Hassan NM, El-Bastawisy ZM, Badran EG (2008) Effect of metribuzin, butachlor and chlorimuron-ethyl on amino acid and protein formation in wheat and maize seedlings. Pestic Biochem Physiol 90:8–18CrossRefGoogle Scholar
  23. Nemat Alla MM, Khedr AA, Serag MM, Abu-Alnaga AZ, Nada RM (2011) Physiological aspects of tolerance in Atriplex halimus L. to NaCl and drought. Acta Physiol Plant 33:547–557CrossRefGoogle Scholar
  24. Plett D, Garnett T, Okamoto M (2017) Molecular genetics to discover and improve nitrogen use efficiency in crop plants. In: Hossain MA, Kamiya T, Burritt DJ, Tran LP, Fujiwara T (eds) Plant macronutrient use efficiency. Academic Press, New York, pp 93–122CrossRefGoogle Scholar
  25. Rhodes D, Hogan AL, Deal L, Jamieson GC, Howorth P (1987) Amino acid metabolism of Lemna minor L. II. Responses to chlorsulfuron. Plant Physiol 84:775–780CrossRefGoogle Scholar
  26. Saja D, Rys M, Stawoska I, Skoczowski A (2016) Metabolic response of cornflower (Centaurea cyanus L.) exposed to tribenuron-methyl: one of the active substances of sulfonylurea herbicides. Acta Physiol Plant 38:168CrossRefGoogle Scholar
  27. Scarcia P, Agrimi G, Germinario L, Ibrahim A, Rottensteiner H, Palmieri F, Palmieri L (2018) In Saccharomyces cerevisiae grown in synthetic minimal medium supplemented with non-fermentable carbon sources glutamate is synthesized within mitochondria. Rend Fis Acc Lincei 29:483–490CrossRefGoogle Scholar
  28. Scarponi L, Nemat Alla MM, Martinetti L (1995) Consequences on nitrogen metabolism in soybean (Glycine max L.) as a result of imazethapyr action on acetohydroxyacid synthase. J Agric Food Chem 43:809CrossRefGoogle Scholar
  29. Singh BK, Stidham MA, Shaner DL (1988) Assay of acetohydroxyacid synthase. Anal Biochem 171:173CrossRefGoogle Scholar
  30. Tian T, Jin Z, Ali B, Guo X, Liu F, Zhang F, Zhang W, He Y, Zhou W (2014) The influence of new herbicide ZJ0273 on the total- and branched-chain amino acids in oilseed rape (Brassica napus L.) leaves as revealed by near-infrared spectroscopy. Acta Physiol Plan 36:2149–2156CrossRefGoogle Scholar
  31. Wani W, Masoodi KZ, Zaid A, Wani SH, Shah F, Meena VS, Wani SA, Mosa KA (2018) Engineering plants for heavy metal stress tolerance. Rend Fis Acc Lincei 29:709–723CrossRefGoogle Scholar
  32. Wray JL, Filner P (1970) Structural and functional relationships of enzyme activities induced by nitrate in barley. Biochem J 119:715–725CrossRefGoogle Scholar
  33. Zhang Y, Xu Y, Wang S, Li X, Zheng M (2017) Resistance mutations of Pro197, Asp376 and Trp574 in the acetohydroxyacid synthase (AHAS) affect pigments, growths, and competitiveness of Descurainia sophia L. Sci Rep 7:16380CrossRefGoogle Scholar

Copyright information

© Accademia Nazionale dei Lincei 2019

Authors and Affiliations

  1. 1.Botany Department, Faculty of ScienceDamietta UniversityNew DamiettaEgypt

Personalised recommendations