Acta Scientific Agriculture (ASAG)(ISSN: 2581-365X)

Research Article Volume 6 Issue 10

Metabolic Processes and Chlorophyll Biosynthesis Affected by Cu and PEG 6000 in Maize

Mamta Hirve and Meeta Jain*

School of Biochemistry, Devi Ahilya University, Takshashila Campus, Indore, India

*Corresponding Author: Meeta Jain, School of Biochemistry, Devi Ahilya University, Takshashila Campus, Indore, India.

Received: May 18, 2022; Published: September 23, 2022


Simultaneous exposure of plants with more than one stresses is a frequent phenomenon in the natural environment. A combination of heavy metals and water deficit stress is one of them. The present study was undertaken to investigate the effects of essential metal copper (Cu), polyethylene glycol (PEG) 6000 induced water deficit and their combination on biochemical attributes and chlorophyll biosynthesis in maize (Zea mays L. cv. Ganga safed-2) seedlings. Results of the study indicate a slight increase in the DNA content of roots with 10% PEG and CuSO4 together. A similar pattern was obtained for total RNA in roots and the effect was substantial and significant with 10% PEG along with 100 µM CuSO4. An increase in protein content and band intensities in protein profile by SDS PAGE was observed, especially, with 100 µM CuSO4, 10% PEG treatment, and its combination with 10 and 100 µM CuSO4. Both the stresses were found to decrease the contents of chlorophylls, carotenoids, and δ-amino levulinic acid (ALA), as well as, ALA synthesizing activity, δ-aminolevulinic acid dehydratase (ALAD), and porphobilinogen deaminase (PBGD) activities. More Cu was accumulated in roots than in shoots. Increased protein content in maize root and shoot tissues by CuSO4, PEG, and their combined treatments may involve the synthesis of new proteins under stress conditions. Unaltered chlorophyll content and enzyme activities involved in chlorophyll biosynthesis by the supply of CuSO4 may be because of balance maintained between formation and utilization of chlorophyll biosynthetic precursors in the presence of essential metal Cu. Inhibition of ALA synthesizing and PBGD activities as well as a decrease in ALA content by PEG, indicates the inhibition of early steps of chlorophyll biosynthetic pathway to be responsible for decreasing the chlorophyll content.

Keywords: Maize; Chlorophyll Biosynthesis; Heavy Metals; Water Deficit


  1. Ali H., et al. “Environmental chemistry and ecotoxicology of hazardous heavy metals: Environmental persistence, toxicity, and bioaccumulation”. Journal of Chemistry (2019): 6730305.
  2. Rodriguez FI., et al. “A copper cofactor for the ethylene receptor ETR1 from Arabidopsis”. Science 283 (1999): 996-998.
  3. Arao T., et al. “Heavy metal contamination of agricultural soil and countermeasures in Japan”. Paddy and Water Environment 8 (2010): 247-257.
  4. Houhou J., et al. “Study of copper phytotoxicity on maize plants (Zea mays ) irrigated by water treated with copper sulfate as algaecide”. International Journal of Science and Research 5 (2016): 1680-1685.
  5. Cobbett C and Goldsbrough P. “Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeo-stasis”. Annual Review of Plant Biology 53 (2002): 159-182.
  6. Demirevska-Kepova K., et al. “Biochemical changes in barely plants after excessive supply of copper and manganese”. Environmental and Experimental Botany 52 (2004): 253-266.
  7. Maksymiec W and Krupa Z. “Effects of methyl jasmonate and excess copper on root and leaf growth”. Biologia Plantarum 51 (2007): 322-326.
  8. Gupta D and Abdullah. “Toxicity of copper and cadmium on germination and seedling growth of maize (Zea mays) Seeds”. Indian Journal of Scientific Research 2 (2011): 67-70.
  9. Contreras R A., et al. “Copper stress induces antioxidant responses and accumulation of sugars and phytochelatins in Antarctic Colobanthus quitensis (Kunth) Bartl”. Biological Research 51 (2018): 48.
  10. Soltani A., et al. “Seed reserve utilization and seedling growth of wheat as affected by drought and salinity”. Environmental and Experimental Botany 55 (2006): 195-200.
  11. Bhatt R M., et al. “Influence of pod load on response of okra to water stress”. Indian Journal of Plant Physiology 10 (2005): 54-59.
  12. Huang B., et al. “Exogenous melatonin alleviates oxidative damages and protects photosystem II in maize seedlings under drought stress”. Frontiers in Plant Science 10 (2019): 677.
  13. Swapna B and Rama Gopal G. “Interactive effects between water stress and heavy metals on seed germination and seedling growth of two green gram (Vigna radiata Wilzec.) cultivars”. Biolife 2 (2014): 291-296.
  14. De Silva N D G., et al. “Effects of combined drought and heavy metal stresses on xylem structure and hydraulic conductivity in red maple (Acer rubrum)”. Journal of Experimental Botany 63 (2012): 5957-5966.
  15. Ku H M., et al. “The effect of water deficit and excess copper on proline metabolism in Nicotiana benthamiana”. Biologia Plantarum 56 (2012): 337-343.
  16. Rucinska-Sobkowiak R. “Water relations in plants subjected to heavy metal stresses”. Acta Physiologiae Plantantarum 38 (2016): 257.
  17. Brzezowski P., et al. “Regulation and function of tetrapyrrole biosynthesis in plants and algae”. Biochimica et Biophysica Acta (BBA)- Bioenergetics 1847 (2015): 968-985.
  18. Beale SI. “Enzymes of chlorophyll biosynthesis”. Photosynthesis Research 60 (1999): 43-73.
  19. Jaffe EK. “The porphobilinogen synthase family of metalloenzymes”. Acta Crystallographica Section D 56 (2000): 115-128.
  20. Beale SI and Weinstein JD. “Tetrapyrrole metabolism in photosynthetic organisms”. In: Dailey (ed.), Biosynthesis of heme and chlorophylls. McGraw-Hill, New York (1990): 287-391.
  21. Vajpayee P., et al. “Chromium (VI) accumulation reduces chlorophyll biosynthesis, nitrate reductase activity and protein content in Nymphaea alba L”. Chemosphere 41 (2000): 1075-1082.
  22. Jain M., et al. “Inhibition of 5-aminolevulinic acid dehydratase activity by arsenic in excised etiolated maize leaf segments during greening”. Journal of Plant Physiology 161 (2004): 251-255.
  23. Sarangthem J., et al. “Inhibition of δ-aminolevulinic acid dehydratase activity by cadmium in excised etiolated maize leaf segments during greening”. Plant Soil and Environment 57 (2011): 332-337.
  24. Gupta P., et al. “Inhibition of 5- aminolevulinic acid dehydratase by mercury in excised greening maize leaf segments”. Plant Physiology and Biochemistry 62 (2013): 63-69.
  25. Dalal V K and Tripathy BC. “Modulation of chlorophyll biosynthesis by water stress in rice seedlings during chloroplast biogenesis”. Plant Cell and Environment 35 (2012): 1685-1703.
  26. Wu Y., et al. “5-Aminolevulinic acid (ALA) biosynthetic and metabolic pathways and its role in higher plants: a review”. Plant Growth Regulation 87 (2019): 357-374.
  27. Marchiol L., et al. “Phytoextraction of heavy metals by canola (Brassica napus) and radish (Raphanus sativus)”. Environmental Pollution 132 (2004): 21-27.
  28. Lowry O H., et al. “Protein measurement with the Folin-Phenol reagent”. Journal of Biological Chemistry 193 (1951): 265-275.
  29. Laemmli UK. “Cleavage of structural proteins during the assembly of the head of bacteriophage T4”. Nature 227 (1970): 680-685.
  30. Webb JM and Levy H B. “New developments in the chemical determination of nucleic acids”. In: Glick D (ed.), Methods of biochemical analysis, InterScience, New York, 6 (1958): 1-30.
  31. Gendimenico GJ., et al. “Diphenylamine-colorimetric method for DNA assay: a shortened procedure by incubating samples at 50ºC”. Analytical Biochemistry 173 (1988): 45-48.
  32. Lichtenthaler H K and Welburn A R. “Determination of total carotenoids and chlorophylls a and b of extracts in different solvents”. Biochemical Society Transactions 11 (1983): 591-592.
  33. Tewari A K and Tripathy BC. “Temperature-stress-induced impairment of chlorophyll biosynthetic reactions in cucumber and wheat”. Plant Physiology 117 (1998): 851-858.
  34. Mauzerall D and Granik S. “The occurrence and determination of δ-aminoleveulinic acid dehydratase and porophobilinogen in urine”. Journal of Biological Chemistry 219 (1956): 435-446.
  35. Prasad DDK and Prasad ARK. “Porphyrin metabolism in lead and mercury treated bajra (Pennisetum typhoideum) seedlings”. Journal of Biosciences 15 (1990): 271-279.
  36. Manivasagaperumal R., et al. “Effect of copper on growth, dry matter yield and nutrient content of Vigna radiata (L.) Wilczek”. Journal of Phytology 3 (2011): 53-62.
  37. Benimeli CS., et al. “Bioaccumulation of copper by Zea mays: Impact on root, shoot and leaf growth”. Water, Air, and Soil Pollution 210 (2010): 365-370.
  38. Dias MAN., et al. “Uptake of seed-applied copper by maize and the effects on seed vigor”. Bragantia Campinas 74 (2015): 241-246.
  39. Ahsan N., et al. “Excess copper induced physiological and proteomic changes in germinating rice seeds”. Chemosphere 67 (2007): 1182-1193.
  40. Amm I., et al. “Protein quality control and elimination of protein waste: The role of the ubiquitin-proteasome system”. Biochimica et Biophysica Acta (BBA)- Molecular Cell Research 1843 (2014): 182-196.
  41. Hasan MK., et al. “Responses of plant proteins to heavy metal stress- A review”. Frontiers in Plant Science 8 (2017): 1492.
  42. Clemens S. “Toxic metal accumulation, responses to exposure and mechanisms of tolerance in plants”. Biochimie 88 (2006): 1707-1719.
  43. Emamverdian A., et al. “Heavy metal stress and some mechanisms of plant defense responses”. The Scientific World Journal 756120 (2015): 18.
  44. Tuna A L., et al. “Maize (Zea mays ) plant responses to excess copper, cadmium, cobalt, lead and chromium”. Fresenius Environmental Bulletin 24 (2015): 3996-4006.
  45. Pochodylo A and Aristilde L. “Molecular dynamics of stability and structures in phytochelatin complexes with Zn, Cu, Fe, Mg, and Ca: Implications for metal detoxification”. Environmental Chemistry Letters 15 (2017): 495-500.
  46. Zhang H., et al. “Proteomic identification of small, copper-responsive proteins in germinating embryos of Oryza sativa”. Annals of Botany 103 (2009): 923-930.
  47. Mohammadkhani N and Heidari R. “Effects of drought stress on soluble proteins in two maize varieties”. Turkish Journal of Biology 32 (2008): 23-30.
  48. Kim S G., et al. “Physiological and proteomic analyses of Korean F1 maize (Zea mays ) hybrids under water-deficit stress during flowering”. The Korean Society for Applied Biological Chemistry 62 (2019): 32.
  49. Godehkahriz SJ., et al. “Effects of water deficit on the physiological response, total protein, and gene expression of Rab17 in wheat (Triticum aestivum)”. Journal of Plant Process and Function 5 (2017): 35-42.
  50. Wang B., et al. “Effects of maize organ-specific drought stress response on yields from transcriptome analysis”. BMC Plant Biology 19 (2019): 335.
  51. Pantola RC., et al. “Copper scavenging potential and its effect on chlorophyll in seedlings of Brassica Juncea (L.) Czern”. Advanced Research Journal of Plant and Animal Sciences 1 (2013): 014-017.
  52. Dey S., et al. “Effect of copper on growth and chlorophyll content in tea plants (Camellia sinensis (L.) O Kuntze”. International Journal of Research in Applied, Natural and Social Sciences 2 (2014): 223-230.
  53. Meher Shivakrishna P., et al. “Effect of PEG-6000 imposed drought stress on RNA content, relative water content (RWC) and chlorophyll content in peanut leaves and roots”. Saudi Journal of Biological Sciences 25 (2018): 285-289.
  54. Shamsi K. “The effects of drought stress on yield, relative water content, proline, soluble carbohydrates and chlorophyll of bread wheat cultivars”. Journal of Animal and Plant Sciences 8 (2010): 1051-1060.
  55. Cha-um S., et al. “Glycinebetaine alleviates water deficit stress in indica rice using proline accumulation, photosynthetic efficiencies, growth performances and yield attributes”. Australian Journal of Crop Science 7 (2013): 213-218.
  56. Chandrasekar V., et al. “Physiological and biochemical responses of hexaploid and tetraploid wheat to drought stress”. Journal of Agronomy and Crop Science 185 (2000): 219-227.
  57. Efeoglu B., et al. “Physiological responses of three maize cultivars to drought stress and recovery”. South African Journal of Botany 75 (2009): 34-42.
  58. Smirnoff N. “Antioxidant systems and plant response to the environment.” In: Environment and plant metabolism: flexibility and acclimation, N Smirnoff (ed.). Bios Scientific Publishers, Oxford, UK (1995): 217-243.


Citation: Mamta Hirve and Meeta Jain. “Metabolic Processes and Chlorophyll Biosynthesis Affected by Cu and PEG 6000 in Maize". Acta Scientific Agriculture 6.10 (2022): 29-40.


Copyright: © 2022 Mamta Hirve and Meeta Jain. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Acceptance rate32%
Acceptance to publication20-30 days
Impact Factor1.014

Indexed In

News and Events

  • Certification for Review
    Acta Scientific certifies the Editors/reviewers for their review done towards the assigned articles of the respective journals.
  • Submission Timeline for Upcoming Issue
    The last date for submission of articles for regular Issues is December 15, 2022.
  • Publication Certificate
    Authors will be issued a "Publication Certificate" as a mark of appreciation for publishing their work.
  • Best Article of the Issue
    The Editors will elect one Best Article after each issue release. The authors of this article will be provided with a certificate of “Best Article of the Issue”.
  • Welcoming Article Submission
    Acta Scientific delightfully welcomes active researchers for submission of articles towards the upcoming issue of respective journals.
  • Contact US