Assessment of Combination of Biocontrol Strains on the Fusaric Acid and other Toxins Secreted from Fusarium oxysporum by HPLC-MS/MS Method and Differential Expression Profiling in Arachis hypogaea L


Affiliations

  • University of Delhi, Department of Botany, Faculty of Sciences, Delhi, 110007, India

Abstract

The ability of Fusarium oxysporum (Schlecht Emend. Snyder and Hansen) in Arachis hypogaea L to produce mycotoxins i.e. Fusaric Acid (FA), Deoxynivalenol (DON), Nivalenol (NIV), Zearalenone, (ZEN), Aflatoxin B1, B2, G1 and G2 in Arachis hypogaea L. leaves in vivo was evaluated in relation to combination of three biocontrol agents, Trichoderma viride + Pseudomonas fluorescens, Trichoderma harzianum + Pseudomonas fluorescens, Trichoderma viride + Trichoderma harzianum. Among the toxins tested, only FA was identified in plants infected with Fusarium oxysporum by LC-MS/MS and quantified using HPLC (4 μg/Kg) Fusaric acid, Deoxynivalenol, Nivalenol, Zearalenone, Aflatoxin B1, B2, G1 and G2 toxins were not detected in plants treated with the combinations of biocontrol agents. The results demonstrate that this procedure is suitable for simultaneous determination of mycotoxins in Fusarium oxysporum of groundnut and the toxin (FA) identified which contributes to the pathogenicity of the fungus during infection. Further differential expression of genes of three leaf samples of control, infected with Fusarium oxysporum and treated leaf sample using combinations of biocontrol agents (Trichoderma viride + Pseudomonas fluorescens ) depicted 5559 genes in control specific, 4316 genes as infected specific and 4264 genes are treated specific. In treated samples 1265 up and 850 down regulation genes were depicted where as in infected sample 605 up and 509 down regulatory genes were depicted. Gene oncology and pathways were found from Uniprot data base. These findings provide new insights into the genetic and biochemical processes required for FA production of Fusarium oxysporum infecting Arachis hypogaea L.

Keywords

Arachis hypogaea L, Fusarium oxysporum, HPLC-MS/MS Method, RNA Transcriptome Sequencing

Subject Discipline

Plant science, Biochemistry,Plant pathology, Biocontrol

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References

Shilman F, Brand Y, Brand A, Hedvat I, Hovav R. Identification and molecular characterization of homeologous Δ9-stearoyl acyl carrier protein desaturase 3 genes from the allotetraploid peanut (Arachis hypogaea). Plant Molecular Biology Rep. 2011; 29:232-–41. https://doi.org/10.1007/s11105-010-0226-9

Fourie G, Steenkamp ET, Ploetz C, Gordon R, Viljoen A. Current status of the taxonomic position of Fusarium oxysporum formae specialis cubense within the Fusarium oxysporum complex. Infection Genetics and Evolut. 2011; 11:533–42. PMid: 21256980. https://doi.org/10.1016/j.meegid.2011.01.012

Joffe AZ. Fusarium species: Their biology and toxicology. New York: John Wiley and Sons; 1986. p. 173.

Miller JH, Harvey HW. Peanut wilt in Georgia. Phytopathol.1932; 2:371–83.

Proctor RH, et al. Evidence that a secondary metabolic biosynthetic gene cluster has grown by gene relocation during evolution of the filamentous fungus Fusarium. Molecular Microbiolo. 2009; 74(5):1128–42. PMid: 19843228. https:// doi.org/10.1111/j.1365-2958.2009.06927.x

Reverberi M, et al. Natural functions of mycotoxins and control of their biosynthesis in fungi. Applied Microbiology and Biotechnol. 2010; 87:899–911. PMid: 20495914. https:// doi.org/10.1007/s00253-010-2657-5

Gaumann E. Fusaric acid as a wilt toxin. Phytopathol. 1957; 47:342–57.

Yabuta T, Kamb K, Hayashi T. Biochemical studies of the ‘bakanae’ fungus of rice. I. Fusarinic acid, a new product of the ‘Bakanae’ fungus. J Agric Chem Soc Jpn. 1937; 10:1059–68.

Bacon CW, Porter JK, Norred WP, Leslie JF. Production of fusaric acid by Fusarium species. Appl Environ Microbio.1996; 62:4039–43. PMid: 8899996 PMCid: PMC168225. https://doi.org/10.1128/AEM.62.11.4039-4043.1996

Notz R, Maurhofer M, Dubach H, Haas D, Defago G. Fusaric acid-producing strains of Fusarium oxysporum alter 2,4-diacetylphloroglucinol biosynthetic gene expression in Pseudomonas fluorescens CHA0 in vitro and biosynthetic gene expression in Pseudomonas fluorescens CHA0 invitro and in the rhizosphere of wheat. Appl Envoron Microbiol. 2002; 68: 2229–35. PMid: 11976092 PMCid: PMC127576.

Schouten A, Vanden Berg G, Edel-Hermann V, Steinberg C, Gautheron N, et.al. Defense responses of Fusarium oxysporum to 2,4-diacetylphloroglucinol, a broad-spectrum antibiotic produced by Pseudomonas fluorescens. Mol Plant Microbe Interact. 2004; 17:1201–11. PMid: 15559985. https://doi.org/10.1094/MPMI.2004.17.11.1201

Kuzniak E, Patykowski J, Urbanek H. Involvement of the antioxidative system in tomato response to fusaric acid treatment. J Phytopathol. 1999; 147:385–90. https://doi.org/10.1111/j.1439-0434.1999.tb03838.x

McLean M. The phytotoxicity of Fusarium metabolites: An update since 1989. Mycopathol.1996; 133:163–79. PMid: 20882471. https://doi.org/10.1007/BF02373024

Lepoivre. Phytopathogie: Bases moleculaires de biologiques des pathsystemes et fondement des strategies de lutte. Brussels, Belgium: De Boeck and Presses Agronomi-ques de Gembloux (Eds.); 2003; 149–67.

Chakrabarti DK, Ghosal S .The disease cycle of mango malformation induced by Fusarium moniliforme var. subglutinans and the curative effects of mangiferin-metal chelates. J Phytopathol.1989; 125:238–46. https://doi.org/10.1111/j.1439-0434.1989.tb01065.x

Gapillout I, Milat ML, Blein JP. Effect of fusaric acid on cells from tomato cultivars resistant or susceptible to Fusarium oxysporum f. sp. lycopersici. Eur J Plant Pathol.1995; 102: 127–32. https://doi.org/10.1007/BF01877099

Kuzniak E. Effect of fusaric acid on reactive oxygen species and antioxidants in tomato cell cultures. J Phytopathol. 2001; 149:575–82. https://doi.org/10.1046/j.1439-0434.2001.00682.x

Pavlovkin J. Effect of fusaric acid on the electrical properties of maize root hairs plasmalemma. Agricultu.1998; 44:350–5.

Bouizgarne B, El-Maarouf-Bouteau H, Frankart C, Reboutier D, Madiona K, Pennarun AM, Monestiez M, Trouverie J, Amiar Z, Briand J. Early physiological responses of Arabidopsis thaliana cells to fusaric acid: toxic and signaling effects. New Phytolog. 2006; 169(1):209. PMid: 16390432. https://doi.org/10.1111/j.14698137.2005.01561.x

Kuo MS. Scheffer RP. Evaluation of fusaric acid as a factor in the development of Fusarium wilt. Phytopathol. 1964; 54:1041–4.

Marre MT,Vergani PFG. Relationship between fusaric acid uptake and its binding to cell structures by leaves of Egeria densa and its toxic effects on membrane permeability and respiration. Physiological and Molecular Plant Pathol.1993; 42:141–57. https://doi.org/10.1006/pmpp.1993.1012

Kohler K, Bentrup FW. The effect of fusaric acid upon electrical membrane properties and ATP level in photoautotrophic cell suspension cultures of Chenopodium rubrum L. Plant Physiol. 1983; 109:355–61. https://doi.org/10.1016/ S0044-328X(83)80117-2

Dalton A, Etherton B. Effects of fusaric acid on tomato root hair membrane potentials and ATP levels. Plant Physio.1984; 74:39–42. PMid: 16663382 PMCid: PMC1066620. https:// doi.org/10.1104/pp.74.1.39

Mehan VK, Bhavanishanker TN, McDonald D. Zearalenone production in groundnut. Trichothecenes and other mycotoxins. Proceedings of the International Mycotoxin Symposium. J. Lacey, ed. Sydney, Australia: John Wiley and Sons Ltd., USA; 1985. p. 73–8.

Pitt JI, Hocking AD. Fungi and food spoilage. 3rd ed. New York: Springer; 2009. PMid: 19067201. https://doi.org/10.1007/978-0-387-92207-2_2

Campbell TC, Stoloff L. Implications of mycotoxins for human health. J Agric Food Chem. 1974; 22:1006–15. PMid: 4372262. https://doi.org/10.1021/jf60196a016

Griffin GJ, Garren KH. Colonization of rye green manure and peanut fruit debris by Aspergillus flavus and Aspergillus niger group in field soils. Appl Environ Microbiol. 1976a; 32: 28–32. PMid: 823865 PMCid: PMC170000. https://doi.org/10.1128/AEM.32.1.28-32.1976

Torres TT, Metta M, Ottenwalder B, Schlotterer C: Gene expression profiling by massively parallel sequencing. Genome Res. 2008; 18:172–7. PMid: 18032722 PMCid: PMC2134766. https://doi.org/10.1101/gr.6984908

Zenoni S, Ferrarini A, Giacomelli E, Xumerle L, Fasoli M, Malerba G, Bellin D, Pezzotti M, Delledonne M. Characterization of transcriptional complexity during berry development in vitis vinifera using RNA-Seq. Plant Physiology. 2010; 152:1787–95. PMid: 20118272 PMCid: PMC2850006. https://doi.org/10.1104/pp.109.149716

Filichkin SA, Priest HD, Givan SA, Shen R, Bryant DW, Fox SE, Wong WK, Mockler TC. Genome-wide mapping of alternative splicing in Arabidopsis thaliana. Genome Res.2010; 20:45–58. PMid: 19858364 PMCid: PMC2798830. https://doi.org/10.1101/gr.093302.109

Vidhyasekaran. Fungal pathogenesis in plants and crops. Molecular biology and host defence mechanisms. New York: Marcel Dekker Inc; 1997. p 553.

Bacon CW, Hinton DM. Fusaric acid and pathogenic interactions of corn and non-corn isolates of Fusarium moniiforme, a nonobligate pathogen of corn. Adv Exp Med Biol. 1996; 392:175–91. PMid: 8850616. https://doi.org/10.1007/978-1-4899-1379-1_16

Stipanovic RD, Puckhaber LS, Liu J, Bell AA. Phytotoxicity of fusaric acid and analogs to cotton. Toxico. 2011; 57:176-8. PMid: 20955724. https://doi.org/10.1016/j.toxicon.2010.10.006

Rajeswari P, Kapoor R. Combined application of different species of Trichoderma enzymes and Pseudomonas fluorescens on the cellulolytic enzymes of Fusarium oxysporum for the control of Fusarium wilt disease in Arachis hypogaea L. Biosciences Biotechnology Research Asi. 2017; 14(3):1169– 76. https://doi.org/10.13005/bbra/2557

Talaviyan JR, Jadeja KB. Efficacy of bioagents alone and in combination microbial population against the wilt incidence of cumin. Journal of Biological Con. 2015; 29(3):162–6. https://doi.org/10.18641/jbc/29/3/86149

Fakhouri W, Walker F, Armbruster W, Buchenauer H. Detoxification of fusaric acid by a nonpathogenic Colletotrichum sp. Physiol Mol Plant Pathol. 2003; 63:263– 9. https://doi.org/10.1016/j.pmpp.2004.03.004

Haas D, Blumer C, Keel C. Biocontrol ability of Xuorescent pseudomonads genetically dissected: Importance of positive feed-back regulation. Biotechnology. 2000; 11:290–7. https://doi.org/10.1016/S0958-1669(00)00098-7

Gamliel A, Katan T, Yunis H, Katan J. Fusarium wilt and crown rot of sweet basil: Involvement of soilborne and airborne inoculum. Phytopathology. 1996; 86:56–62. https:// doi.org/10.1094/Phyto-86-56

Notz R, Maurhofer M, Dubach H, Haas D, Defago G. Fusaric acid-producing strains of Fusarium oxysporum alter 2,4-diac-etylphloroglucinol biosynthetic gene expression in Pseudomonas fluorescens CHA0 in vitro and in the rhizosphere of wheat. Appl Environ Microbiol. 2002; 68:2229–35. PMid: 11976092 PMCid: PMC127576. https:// doi.org/10.1128/AEM.68.5.2229-2235.2002


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