Influence of eco-friendly phytotoxic metabolites from Lasiodiplodia pseudotheobromae C1136 on physiological, biochemical, and ultrastructural changes on tested weeds

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In this study, the active metabolites from both the wild strain of Lasiodiplodia pseudotheobromae C1136 and three genetically enhanced strains of C1136 were obtained through random mutagenesis. The effect of the active metabolites from these strains was evaluated in relation to physiological, biochemical, and ultrastructural changes on the leaves of two weeds (Amaranthus hybridus and Echinochloa crus-galli). The phytotoxic metabolites secreted by the genetically enhanced strains showed a decrease in the pigments (chl a, chl b, and carotenoids), carbohydrate content, and the amino acid profile. On the other hand, an increase in total phenols of the tested leaves was observed when compared with the untreated leaves. The scanning electron microscopy showed the presence of damages, necrosis, degradation, and ultrastructural changes on the tested leaf tissues of the weeds. Also, increased lipid peroxidation and electrolyte leakage were also observed on the tested weeds treated with phytotoxic metabolites secreted by the genetically enhanced strains. We also showed that the phytotoxins from the strains of C1136 are biocompatible and that it improved soil CO2 evolution, organic carbon content, and enzymatic activity (acidic and alkaline phosphatase, dehydrogenases, cellulase, catalase). The study validates the severe pathological effects of phytotoxic metabolites from the strains of C1136 on the leaves of the weeds presented in this study. The mode of action of the phytotoxic metabolites produced from this bioherbicidal isolates will go a long way in preventing environmental hazards.

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  1. Adetunji CO, Oloke JK (2013) Efficacy of freshly prepared pesta granular formulations from the multi-combination of wild and mutant strain of Lasiodiplodia pseudotheobromae and Pseudomonas aeruginosa. Agric Univ Tirana 12:555–563

  2. Adetunji C, Oloke J, Kumar A, Swaranjit S, Akpor B (2017a) Synergetic effect of rhamnolipid from Pseudomonas aeruginosa C1501 and phytotoxic metabolite from Lasiodiplodia pseudotheobromae C1136 on Amaranthus hybridus L. and Echinochloa crus-galli weeds. Environ Sci Pollut Res 24:13700–13709.

  3. Adetunji CO, Oloke JK, Prasad G, Bello OM, Osemwegie OO, Mishra P, Jolly RS (2017b) Isolation, identification, characterization and screening of rhizospheric bacteria for herbicidal activity. 8:195–205

  4. Adetunji CO, Oloke JK, Osemwegie OO (2018) Environmental fate and effects of granular pesta formulation from strains of Pseudomonas aeruginosa C1501 and Lasiodiplodia pseudotheobromae C1136 on soil activity and weeds. Chemosphere 195(2018):98–107.

  5. Aebi H (1984) Catalase. In: Packer L (ed) Methods in Enzymology, vol 105. Academic Press, Orlando, pp 121–126

  6. Aldesuquy HS, Baka ZAM (1992) Physiological and biochemical changes in host leaf tissues associated with the growth of two biotrophic fungi growing in Egypt. Phyton 32(1):129–142

  7. Allison SD, Weintraub MN, Gartner TB, Waldrop MP (2010) Evolutionary-economic principles as regulators of soil enzyme production and ecosystem function. Springer, Berlin

  8. Anderson M (1996) Glutathione. In: Punchard NA, Kelly FJ (eds) Free Radicals, a Practical Approach. Oxford University Press, New York, p 213

  9. Baka ZM, Aldesuquy HS (1992) Studies on the powdery mildew fungus Sphaerotheca fuliginea: ultrastructure and host responses. Acta Bot Hung 37:15–31

  10. Baker B, Zambryski P, Staskawicz B, Dinesh-Kumar SP (1997) Signaling in plant-microbe interactions. Science 276:726–733

  11. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287.

  12. Berestetskiy AO (2008) A review of fungal phytotoxins: from basic studies to practical use. Appl Biochem Microbiol 44:453–465

  13. Bo AB, Kim JD, Kim YS, Sin HT, Kim HJ, Khaitov B, Ko YK, Park KW, Choi JS (2019) Isolation, identification and characterization of Streptomyces metabolites as a potential bioherbicide. PLoS One 14(9):e0222933.

  14. Böger P (2003) Mode of action for chloroacetamides and functionally related compounds. J Pestic Sci 28:324–329

  15. Bunkers GJ, Strobel GA (1991) A proposed mode of action for green island induction by the eremophilane phytotoxins produced by Drechslera gigantea. Physiol Mol Plant Pathol 38:313–323

  16. Chacon-Morales PO, Amara-Luis JM, Bahsas A (2013) Isolation and characterization of (+)- mellein, the first isocoumarin reported in Stevia genus. Adv Quim 8(3):145–151

  17. Chun JC, Kim JC, Hwang IT, Kim SE (2002) Acteoside from Rehmannia glutinosa nullifies paraquat activity in Cucumis sativus. Pest Biochem Physiol 72:153–159

  18. Daguerre Y, Siegel K, Edel-Hermann V, Steinberg C (2014) Fungal proteins and genes associated with biocontrol mechanisms of soil-borne pathogens: a review. Fungal Biol Rev 28:97–125

  19. Dan K, Nagata M, Yamashita I (1996) Changes in lipid peroxidation and antioxidants content in cotyledons of Japanese radish. J Jap Soc Hort Sci 65:603–608

  20. Dayan FE, Watson SB (2011) Plant cell membrane as a marker for light-dependent and light-independent herbicide mechanisms of action. Pestic Biochem Physiol 101:182–190.

  21. Dayan FE, Cantrell CL, Duke SO (2009) Natural products in crop protection. Bioorg Med Chem 17:4022–4034

  22. Duke SO, Kenyon WH (1993) Peroxidizing activity determined by cellular leakage. In: Böger P, Sandmann G (eds) Target assays for modern herbicides and related compounds. Lewis Publishers, Boca Raton, pp 61–66

  23. El-Sayed W (2005) Biological control of weeds with pathogens: currents status and future trends. J Plant Dis Protect 112:209–221

  24. Evidente A, Motta A (2001) Phytotoxins from fungi, pathogenic, for Agrarian, forestal and weedy plants. In: Tringali C (ed) Bioactive compound natural resources. Taylor and Francis, London, pp 474–525

  25. Galea V (2013) Australia’s first bioherbicide approaching release.

  26. Ghorbani R, Leifert C, Seel W (2005) 2005. Biological control of weeds with antagonistic plant pathogens. Adv. Agron. 86:191–225

  27. Harding DP, Raizada MN (2015) Controlling weeds with fingi, bacteria and viruses: a review. Front Plant Sci 6:659.

  28. Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta. 207(4):604–611

  29. Humbeck K, Quast S, Krupinska K (1996) Functional and molecular changes in the photosynthetic apparatus during senescence of flag leaves from field-grown barley plants. Plant Cell Environ 19:337–344

  30. Hüter OF (2010) Use of natural products in the crop protection industry. Phytochem Rev 10:185–194

  31. Jia X, Zhao Y, Wang W, He Y (2015) Elevated temperature altered photosynthetic products in wheat seedlings and organic compounds and biological activity in rhizopshere soil under cadmium stress. Sci Rep 5:14426

  32. Jing L, Howard AC (2010) Biotechnol Lett Springer Verlag 32(5):601–608

  33. Kandeler E, Tscherko D, Bardgett RD, Hobbs PJ, Kamplichler C, Jones TH (1998) The response of soil microorganisms and roots to elevated CO2 and temperature in a terrestrial model ecosystem. Plant Soil 202:251–262

  34. Lim PO, Woo HR, Nam HG (2003) Molecular genetics of leaf senescence in Arabidopsis. Trends Plant Sci 8:272–278

  35. Lincoln JE, Richael C, Overduin B, Smith K, Bostock R, Gilchrist DG (2002) Expression of the antiapoptotic baculovirus p35 gene in tomato blocks programmed cell death and provides broad-spectrum resistance to disease. Proc Natl Acad Sci U S A 99:15217–15221

  36. Lu C, Zhang J (1998a) Changes in photosystem II function during senescence of wheat leaves. Physiol Plant 104:239–247

  37. Lu C, Zhang J (1998b) Modifications in photosystem II photochemistry in senescent leaves of maize plants. J Exp Bot 49:1671–1679

  38. Ma Y, Li J, Wu, Juan K, Zhaoyu F, Larry M, Ding X, Ge G, Wu L (2018) Bacterial and fungal community composition and functional activity associated with lake wetland water level gradients. Sci Rep 8:760.

  39. Mascher R, Nagy E, Lippmann B, Hörnlein S, Fischer S, Scheiding W, Neagoe A, Bergmann H (2005) Improvement of tolerance to paraquat and drought in barley (Hordeum vulgare L.) by exogenous 2-aminoethanol: effects on superoxide dismutase activity and chloroplast ultrastructure. Plant Sci 168:691–698

  40. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880

  41. Nicholson RL (1992) Phenolic compounds and their role in disease resistance. Annu Rev Phytopathol 30:369–389

  42. Pedras MSC, Zaharia IL, Gai Y, Zhou Y, Ward DE (2001) In planta sequential hydroxylation and glycosylation of a fungal phytotoxin: avoiding cell death and overcoming the fungal invader. Proc Natl Acad Sci U S A 98:747–752

  43. Prashar P, Kapoor N, Sachdeva S (2014) Rhizosphere: its structure, bacterial diversity and significance. Rev Environ Sci Biotechnol 13:63–77

  44. Rajasekharan R, Rodrigues R, Reddy SP (2001) Herbicides comprising phytotoxins of Lasiodiplodia theobromae, their production and use. International patent no: PCT/IN2000/000028.

  45. Ricardo SA, Erick AE, Ramon SR, Tatiana CP, Veronica OA, Monica QF, Marcia CS, Anderson SS, Adriana COS, Adriano GC (2018) Physical hazards in dairy products: incidence in a consumer complaint website in Brazil. Food Control 86(2018):66–70.

  46. Scholes JD, Farrar JF (1985) Photosynthesis and chloroplast functioning within individual pustules of Uromyces muscari on bluebell leaves. Physiol Plant Pathol 27:387–400

  47. Shabana YM, Baka ZA, Abdel-Fattah GM (1997) Alternaria eichhorniae, a biological control agent for water hyacinth: mycoherbicidal formulation and physiological and ultrastructural host responses. Eur J Plant Pathol 103:99–111

  48. Stergiopoulos I, Collemare J, Mehrabi R, De Wit P (2013) Phytotoxic secondary metabolites and peptides produced by plant pathogenic Dothideomycete fungi. Fems Microbiol Rev 37:67–93.

  49. Tabatabai MA (1994) Soil enzymes, in Methods of Soil Analysis. Part 2-Chemical and Microbiological Properties. In: Page AL, Miller RH, Keeney DR (eds) Soil Science Society of America, Madison Wis USA, pp 775–883

  50. Tanaka T, Abbas HK, Duke SO (1993) Structure-dependent phytotoxicity of fumonisins and related compounds in a duckweed bioassay. Phytochemistry. 33:779–785

  51. Tashiro T, Flukuda Y, Osawa T, Namiki M (2007) Oil and minor components of sesame (Sesamum indicum L.) strains. J Am Oil Chem Soc 67(8):508–511

  52. Velikova V (2000) Oxidative stress and some antioxidant systems in acid rain- treated bean plants: protective role of exogenous polyamines. Plant Sci 151:59–66.

  53. Walton JD (1996) Host-selective toxins: agents of compatibility. Plant Cell 8:1723–1733.

  54. Yang L, Li T, Li F, Lemcoff JH, Cohen S (2008) Fertilization regulates soil enzymatic activity and fertility dynamics in a cucumber field. Sci Hortic 116:21–26

  55. Yi X, Jing D, Wan J, Ma Y, Wang Y (2016) Temporal and spatial variations of contaminant removal, enzyme activities, and microbial community structure in a pilot horizontal subsurface flow constructed wetland purifying industrial run off. Environ Sci Pollut Res 23:1–12

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Special thanks to Daniel Hefft, School of Chemical Engineering (University of Birmingham, UK).

Funding information

The authors are grateful to the Council of Scientific and Industrial Research (CSIR), New Delhi, India and The World Academy of Science (TWAS), Italy for providing the necessary facilities and opportunity to carry out the molecular, biochemical as well as the scanning electron microscopy analysis of this work at Institute of Microbial Technology, Sector 39A, Chandigarh, India, Department of Molecular Biology and Microbial Type Culture Collection and Gene Bank (MTCC) under the supervision of Dr. Prasad Gandham and Dr. R.S Jolly. FR number: 3240267282. (2013).

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Correspondence to Charles Oluwaseun Adetunji.

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Adetunji, C.O., Oloke, J.K., Phazang, P. et al. Influence of eco-friendly phytotoxic metabolites from Lasiodiplodia pseudotheobromae C1136 on physiological, biochemical, and ultrastructural changes on tested weeds. Environ Sci Pollut Res (2020).

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  • Microorganism
  • Soil activity
  • Chlorophyll content
  • Enzymatic activity
  • Bioherbicides
  • Random mutagenesis