Comparative Analysis of Determination Methods of Glyphosate Degradation by Trichoderma asperellum Strain JK-28: A Multivariate Statistical Approach

Main Article Content

Atim Asitok
Maurice Ekpenyong

Abstract

Aims: This study aimed at demonstrating the ability of a mold species, isolated from a rice farm agricultural soil, chronically treated with glyphosate herbicide, to degrade glyphosate and to compare different evaluation methods of biodegradation.


Study Design: A completely randomized design of a factorial experiment was used.


Place and Duration of Study:    The study was conducted at the Department of Microbiology, University of Calabar, Nigeria between February and April.


Methodology: Molds were isolated from nine rice agricultural soil samples using potato dextrose agar. The molds were screened for glyphosate degradation using different media to determine their requirement for supplementation or not at all. A completely randomized design of a factorial experiment involving 3 factors at 3 levels each, with three replications, was adopted for glyphosate degradation studies. Degradation was monitored by dehydrogenase activity, carbon dioxide evolution, pH changes, fungal dry weight and residual glyphosate determination. A comparative evaluation of the different methods was made using multivariate statistics.


Results: The mold, identified as Trichoderma asperellum strain JK-28, was incapable of using glyphosate as sole sources of phosphorus and nitrogen and so required nutrient supplementation. Largest significant correlation between dependent variables (P < .001; r = - 779) occurred between dehydrogenase activity and fungal dry weight while the lowest was between residual glyphosate and fungal dry weight. Residual glyphosate and pH had negative correlations with other dependent variables. Principal component analysis extracted only one component with a cumulative variance of 70%. Multivariate analysis of variance (three-way MANOVA) of data, interpreted from the Pillai’s Trace test, was significant at P <.001 for the main, two- and three-way interactions.


Conclusion: Trichoderma asperellum strain JK-28 could degrade glyphosate using it as sole carbon source but with nutrient supplementation. Residual glyphosate determination is recommended as most reliable method for evaluating organic pollutant biodegradation.

Keywords:
Trichoderma asperellum strain JK-28, glyphosate, degradation, correlation analysis, principal component analysis, three-way MANOVA

Article Details

How to Cite
Asitok, A., & Ekpenyong, M. (2019). Comparative Analysis of Determination Methods of Glyphosate Degradation by Trichoderma asperellum Strain JK-28: A Multivariate Statistical Approach. Journal of Agriculture and Ecology Research International, 19(1), 1-14. https://doi.org/10.9734/jaeri/2019/v19i130070
Section
Original Research Article

References

Cox C. Glyphosate, Part I: Toxicology. J Pesticide Reform. 1995;15:14-20.

Zablotowicz RM, Reddy KN. Impact of glyphosate on Bradyrhizobium japonicum symbiosis with glyphosate-resistant transgenic soybean: A minireview. J Environ Qual 2004;33:825-833.

Available:https://doi.org/10.2134/jeq2004.0825

Peñaloza-Vazquez A, Mena GL, Herrera-Estrella L, Bailey AM. Cloning and sequencing of the genes involved in glyphosate utilization by Pseudomonas pseudomallei. Appl Environ Microbiol. 1995;61:538-543.

Cox C. Glyphosate, Part II: Human exposure and ecological effects. J Pesticide Reform. 1995;15:14-20.

Gimsing AL, Borggaard OK, Bang M. Influence of soil composition on adsorption of glyphosate and phosphate by contrasting Danish surface soils. Eur J Soil Sci 2004;55:183-191.

Available:https://doi.org/10.1046/j.1365-2389.2003.00585.x

Yu XM, Yu T, Yin GH, Dong QL, An M, Wang HR, Ai CX. Glyphosate biodegradation and potential soil bioremediation by Bacillus subtilis strain Bs-15. Genet Mol Res 2015; 14: 14717-14730. Available:https://doi.org/10.4238/2015.November.18.37

Haney RL, Senseman SA, Hons FM, Zuberer DA. Effect of glyphosate on soil microbial activity and biomass. Weed Sci 2000;48:89-93.

Available:https://doi.org/10.1614/0043-1745(2000)048[0089:EOGOSM]2.0.CO;2

Ekpenyong MG, Antai SP, Essien JP. Quantitative and qualitative assessment of hydrocarbon degrading bacteria and fungi in Qua Iboe Estuary, Nigeria. Res J Microbiol. 2007;2:415-425.

Available:https://doi.org/10.3923/jm.2007.415.425

Moneke AN, Okpala GN, Anyanwu CU. Biodegradation of glyphosate herbicide in vitro using bacterial isolates from four rice fields. Afr J Biotechnol. 2010;9:4067-4074.

Ekpenyong MG, Antai SP, Asitok AD, Ekpo BO. Plackett-Burman design and response surface optimization of medium trace nutrients for glycolipopeptide biosurfactant production. Iranian Biomed J. 2017;21:249-260. Available:https://doi.org/10.18869/acadpub.ibj.21.4.249

Manter DK, Vivanco JM. Use of the ITS primers, ITS1-F and ITS4, to characterize fungal abundance and diversity in mixed template samples by qPCR and length heterogeneity analysis. J Microbiol Meth 2007;71:7-14. Available:https://doi.org/10.1016/j.mimet.2007.06.016

Jan MR, Shah J, Muhammad M, Ara B. Glyphosate herbicide residue determination in samples of environmental importance using spectrophotometric method. J Hazard Mater. 2009;169:742-745. Available:https://doi.org/10.1016/j.jhazmat.2009.04.003

Casida Jr LE, Klein DD, Santoro T. Soil dehydrogenase activity. Soil Sci. 1964; 98: 371-376.

Available:https://doi.org/10.1097/00010694-196412000-00004

Asitok A, Ekpenyong M, Antai S. Multivariate statistics of fertility parameter fluxes in cement-dust-polluted soils in Mfamosing, Nigeria; impact on agriculture. J Microbiol Biotechnol Food Sci. 2019;8(6). (In press).

Ng HE, Rag SSA, Wong SH, Tey D, Tan HM. Estimation of fungal growth using the ergosterol assay: A rapid tool in assessing the microbiological status of grains and feeds. Lett Appl Microbiol. 2007;46:113-118.

Available:https://doi.org/10.1111/j.1472-765X.2007.02279.x

Ekpenyong MG, Antai SP. Influence of pH on cadmium toxicity to species of Bacillus (02 and 12) during biodegradation of crude oil. Int J Biol Chem. 2007;1:29-37.

Available:https://doi.org/10.3923/ijbc.2007.29.37

Asitok AD, Antai SP, Ekpenyong MG. Water soluble fraction of crude oil uncouples protease biosynthesis and activity in hydrocarbonoclastic bacteria; Implications for Natural Attenuation. Int J Sci. 2017;6:5-21.

Wu Q, Sun R, Ni M, Yu J, Li Y, Yu C, Dou K, Ren J, Chen J. Identification ovel fungus Trichoderma asperellum GDFS1009 and comprehensive evaluation of its biocontrol efficacy. PloS One. 2017;12:e0178857. Available:https://doi.org/10.1371/journal.pone.0179957

Benson NU, Asuquo FE, Williams AB, Essien JP, Ekong CI, Akpabio O, Olajireo AA. Source evaluation and trace metal contamination in benthic sediments from equatorial ecosystems using multivariate statistical techniques. PLoS One. 2016; 11:e0156485.

Available:https://doi.org/10.1371/journal.pone.0156485

Asitok AD, Ekpenyong MG. Statistical distribution of halotolerant/halophilic biosurfactant-producing yeasts in Qua Iboe estuarine sediments. Int J Sci. 2019;8(5). (In press).

Wolinska A, Stepniewska Z. Dehydrogenase activity in the soil environment. In. Canuto RA. (ed.) Dehydrogenases. Rijeka: Intech Publisher; 2012.

Available:https://doi.org/10.5772/48294

Salazar S, Sanchez L, Alvarez J, Valverde A, Galindo P, Igual J, Peix A, Santa-Regina I. Correlation among soil enzyme activities under different forest system management practices. Ecol Eng. 2011; 37:1123-1131. Available:https://doi.org/10.1016/j.ecoleng.2011.02.007

Itah AY, Essien JP. Growth profile and hydrocarbonoclastic potential of microorganisms isolated from tarballs in the Bight of Bonny, Nigeria. World J Microbiol Biotechnol. 2005;21:1317-1322. Available:https://doi.org/10.1007/s11274-004-6694-z

Tabachnick BG, Fidell LS. Using multivariate statistics. New York: Harper Collins College Publishers; 1996.