Antifungal Activity of Endophytic Bacteria Associated with Sweet Sorghum (Sorghum bicolor)

Authors

  • Siti Meliah Research Center for Biology, Indonesian Institute of Science (LIPI), Jalan Raya Jakarta Bogor Km. 46 Cibinong 16911, West Java
  • Tri Ratna Sulistiyani Research Center for Biology, Indonesian Institute of Science (LIPI), Jalan Raya Jakarta Bogor Km. 46 Cibinong 16911, West Java
  • Puspita Lisdiyanti Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Jalan Raya Jakarta Bogor Km. 46 Cibinong 16911, West Java
  • Atit Kanti Research Center for Biology, Indonesian Institute of Science (LIPI), Jalan Raya Jakarta Bogor Km. 46 Cibinong 16911, West Java
  • I Made Sudiana Research Center for Biology, Indonesian Institute of Science (LIPI), Jalan Raya Jakarta Bogor Km. 46 Cibinong 16911, West Java
  • Masaru Kobayashi Graduate School of Agriculture, Division of Applied Life Sciences, Kitashirakawa, Oiwake-cho, Sakyo-ku, Kyoto 606-8502

DOI:

https://doi.org/10.5614/j.math.fund.sci.2021.53.1.2

Keywords:

antifungal, bacteria, endophytes, enzymes, mechanisms

Abstract

The contribution of endophytic bacteria to the wellbeing of plants as biocontrol agents may be due to endophytic bacteria growing in the same niche as phytopathogens. This work was conducted to study the antagonistic activity of endophytic bacteria recovered from sweet sorghum against Sclerotium rolfsii, Fusarium solani, Fusarium oxysporum, Colletotrichum gloeosporioides in vitro and evaluate the mechanisms of these fungal inhibitions. We selected 78 endophytic bacteria from the stem and root of sweet sorghum plants. They were tested for antagonist activity by direct confrontation method. Antifungal compound production and lytic enzyme activity were examined to determine their mechanisms in inhibiting fungal pathogens. Antifungal compound production was checked by detecting the presence of NRPS and PKS genes. Lytic enzyme activity of the bacteria was evaluated by their ability to produce cellulase, chitinase, and protease. Selected bacteria were identified using molecular analysis based on the 16S rRNA gene. 14 out of the 78 tested isolates showed antagonistic activity and two were able to inhibit all four tested fungal strains. Four bacteria, designated as ACIL1, ACNM4, ACNM6, and ATNM4, produced natural products via NRPS pathway, but only one bacterial extract, designated as ACNM4, showed fungal inhibition. Ten isolates were able to produce hydrolytic enzymes. Endophytic bacteria identified as Burkholderia were revealed to have potential as a biocontrol agent.

References

Yu, H., Zhang, L., Li, L., Zheng, C., Guo, L., Li, W., Sun, P. & Qin, L., Recent Developments and Future Prospect of Antimicrobial Metabolites Produced by Endophytes, Microbiological Research, 165, pp. 437-449, 2010.

Compant, S., Clement, C. & Sessitsch, A., Plant Growth-promoting Bacteria in the Rhizo- and Endosphere of Plants: Their Role, Colonization, Mechanisms Involved and Prospects for Utilization, Soil Biology and Biochemistry, 42, pp. 669-678, 2010.

Tan, R.X. & Zou, W.X., Endophytes: A Rich Source of Functional Metabolites, Natural Product Reports, 18, pp. 448-459, 2001.

Mano, H. & Morisaki, H., Endophytic Bacteria in the Rice Plant, Microbes and Environments, 23, pp. 107-117, 2008

Ruby, E.J., A Review: Bacterial Endophytes and Their Bioprospecting, Journal of Pharmacy Research, 4, pp. 795-799, 2011.

Mendes, R., Pizzirani-Kleiner, A.A., Araujo, W.L. & Raaijmakers, J.M., Diversity of Cultivated Endophytic Bacteria from Sugarcane: Genetic and Biochemical Characterization of Burkhloderia cepacia Complex Isolates, Applied and Environmental Microbiology, 73, pp. 7259-7267, 2007.

Chen, C., Bauske, E.M., Musson, G., Rodrigues-Kabana, R. & Koepper, J.W., Biological Control of Fusarium Wilt on Cotton by Use of Endophytic Bacteria, Biological Control, 5, pp. 83-91, 1995.

Cho, K.M., Hong, S.Y., Lee, S.M., Kim, Y.H., Kahng, G.G., Lim, Y.P., Kim, H. & Yun, H.D., Endophytic Bacteria Communities in Ginseng and Their Antifungal Activity Against Pathogens, Microbial Ecology, 54, pp. 341-351, 2007.

Suriani & Muis, A., Fusarium spp. on Maize and Its Control with Utilizing Endophytic Microbes, Iptek Tanaman Pangan, 11, pp. 133-142, 2016.

Pinaria, A.G., Liew, E.C.Y., & Burgess, L.W., Fusarium Species Associated with Vanilla Stem Rot in Indonesia, Australasian Plant Pathology, 39, pp. 176-183, 2010.

Mostert, D., Molina, A.B., Daniells, J., Fourie, G., Hermanto, C., Chao, C-P., et al., The Distribution and Host Range of the Banana Fusarium Wilt Fungus, Fusarium oxysporum f. sp. cubense in Asia, PLOS ONE, 12(7):e0181630, 2017.

Soenartiningsih, Aqil, M. & Andayani, N.N., Strategy for Controlling Fusarium sp. and Mycotoxin Contamination in Corn, Iptek Tanaman Pangan, 11, pp. 85-93, 2016.

Vigier, B., Reid, L.M., Dwyer, L.M., Stewart, D.W., Sinha, R.C., Arnason, J.T. & Butler, G., Maize Resistance to Gibberella Ear Rot Symptoms, Deoxynivalenol and Yield, Canadian Journal of Plant Pathology, 23, pp. 99-105, 2001.

Compant, S., Duffy, B., Nowak, J., Clent, C. & Barka E.A., Use of Plant Growth-Promoting Bacteria Biocontrol of Plant Disease: Principles, Mechanisms of Action and Future Prospects. Applied and Environmental Microbiology, 71(9), pp. 4951, 2005.

Khabbaz, S.E., Zhang, L., Caceres, L.A., Sumarah, M., Wang, A. & Abbasi, P.A., Characterization of Antagonistic Bacillus and Pseudomonas Strain for Biocontrol Potential and Suppression of Damping-off and Root Rot Diseases. Annals of Applied Biology, 166, pp. 456-471, 2015.

Shafi, J., Tian, H. & Ji, M., Bacillus species as Versatile Weapons for Plant Pathogens: A Review, Biotechnology & Biotechnological Equipment, 31(3), 446-459, 2017. DOI:10.1080/13102818.2017.1286950.

Widyatmoko, D., Sudiana, I.M., Lestari, R., Ariati, S.R., Lisdiyanti, P., Munawar, & S.S., Nugroho, S.(Ed), Isolation and Characterization of Nitrogen Fixing Endophytic Bacteria Associated with Sweet Sorghum (Sorghum bicolor), Proceeding of the 1st SATREPS conference., pp. 110-117, 2017.

Amos, G.C.A., Borsetto, C., Laskaris, P., Krsek, M., Berry, A.E., Newsham, K.K., Calvo-Bado, L., Pearce, D.A., Vallin C. & Wellington, E.M.H., Designing and Implementing an Assay for the Detection of Rare and Divergent NRPS and PKS Clones in European, Antarctic and Cuban Soils, PLOS ONE, 10, pp. 1-15, 2015.

Liang, Y.L., Zhang, Z., Wu, M., Wu, Y. & Feng, J.X., Isolation, Screening and Identification of Cellulolytic Bacteria from Natural Reserves in Subtropical Region of China and Optimization of Cellulase Production by Paenibacillus terrae ME27-1, BioMed Research International, pp. 1-13, 2014.

Kiran, T., Asad, W., Siddiqui, S., Ajaz, M. & Rassol, S.A., Industrially Important Hydrolytic Enzyme Diversity Explored in Stove Ash Bacterial Isolates, Pakistan Journal of Pharmaceutical Sciences, 28, pp. 2035-2040, 2015.

Lane, D.J., Nucleic Acid Techniques in Bacterial Systematics, Stackebrandt, E. & Goodfellow, M., (Ed.) Wiley, 1991.

Hall, T.A., BioEdit: A User-Friendly Biological Sequence Alignment Editor and Analysis Program for Windows 95/98/NT, Nucleic Acids Symposium Series, 41, pp. 95-98, 1999.

Yoon, S.H., Ha, S.M., Kwon, S., Lim, J., Kim, Y., Seo, H. & Chun, J., Introducing EzBioCloud: A Taxonomically United Database of 16S rRNA and Whole Genome Assemblies, International Journal of Systematic and Evolutionary Microbiology, 67, pp. 1613-1617, 2017.

Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K., MEGA X: Molecular Evolutionary Genetic Analysis Across Computing Platforms, Molecular Biology and Evolution, 35, pp. 1547-1549, 2018.

Edgar, R.C., MUSCLE: Multiple Sequences Alignment with High Accuracy and High Throughput, Nucleic Acids Research, 32, pp. 1792-1797, 2004.

Satou, N. & Nei, M., The Neighbor-Joining Method: A New Methods for Reconstructing Phylogenetic Trees, Molecular Biology and Evolution, 4, pp. 406-425, 1987.

Kimura, M., A Simple Method for Estimating Evolutionary Rate of Base Substitutions Through Comparative Studies of Nucleotide Sequences, Journal of Molecular Evolution, 16, pp. 111-120, 1980.

Abdulsalaam, S. & Shenge, K.C., Seed Borne Pathogens on Farmer-Saved Sorghum (Sorghum bicolor L.) Seeds, Journal of Stored Products and Postharvest Research, 2, pp. 24-28, 2011.

Donadio, S., Monciardini, P. & Sosio, M., Polyketide Synthases and Nonribosomal Peptide Synthetases: The Emerging View from Bacterial Genomics, Natural Product Reports, 24, pp. 1073-1109, 2007.

Strieker, M., Tanovi?, A. & Marahiel, M.A., Nonribosomal peptide Synthehases: Structures and Dinamics, Current Opinion in Structural Biology, 20, pp. 234-240, 2010.

Wang, H., Fewer, D.P., Holm, L., Rouhiainen, L. & Sivonen, K., Atlas of Nonribosomal Peptide and Polyketide Biosynthetic Pathways Reveals Common Occurrence of Nonmodular Enzymes, PNAS, 111(25), pp. 9259-9264, 2014.

Awan, A.R., Blount, B.A., Bell, D.J., Shaw, W.M., Ho, J.C.H., McKiernan, R.M. & Ellis, T., Biosynthesis of Antibiotic Nonribosomal Peptide Penicillin in Baker?s Yeast, Nature Communications, 8, 15202, 2017. DOI:10.1038/ncomms15202.

Felnagle, E.A., Jackson, E.E., Chan, Y.A., Podevels, A.M., Berti, A.D., McMahon, M.D. & Thomas, M.G., Nonribosomal Peptide Synthetases Involved in the Production of Medically Relevant Natural Products, Molecular Pharmaceutics, 5(2), pp. 191-211, 2008.

Adam, D.J., Fungal Cell Wall Chitinase and Glucanases, Microbiology, 150, pp. 2029-2035, 2004.

Fesel, P.H. & Zuccaro, A., ?-glucan: Crucial Component of the Fungal Cell Wall and Elusive MAMP in Plants, Fungal Genetics and Biology, 90, pp. 53-60, 2016.

Ordentlich, A., Elad, Y. & Chet, I., The Role of Chitinase of Serratia marcescens in Biocontrol of Sclerotium rolfsii, Ecology and Epidemiology, 78, pp. 84-88, 1988.

Lim, H.S., Kim, Y.S. & Kim, S.D., Pseudomonas stutzeri YPL-1 Genetic Transformation and Antifungal Mechanism Against Fusarium solani, An Agent of Plant Root Rot, Applied and Environmental Microbiology, 57, pp. 510-516, 1991.

Van den Burg, H.A., Harrison, S.J., Joosten, M.H., Vervoot, J. & de Wit P.J., Cladosporium fulvum Avr4 Protects Fungal Cell Walls Against Hydrolysis by Plant Chitinases Accumulating During Infection, Molecular Plant Microbe Interaction, 19, pp. 1420-1430, 2006.

Trevors, J.T., One Gram of Soil: A Microbial Biochemical Gene Library, Antonie van Leeuwenhoek, 97, pp. 99-106, 2010.

Mareque, C., Taul C., Beracochea, M. & Battistoni, F., Isolation, Characterization, and Plant Growth Promotion Effect of Putative Bacterial Endophytes Associated with Sweet Sorghum (Sorghum bicolor (L) Moench), Annals of Microbiology, 65, pp. 1057-1067, 2015.

Caballero-Mellado, J., Martinez-Aguilar, L., Paredes-Valdez, G. & Estrada-de los Santos, P., Burkholderia unamae sp. nov., an N2-fixing rhizospheric and endophytic species, International Journal of Systematic and Evolutionary Microbiology, 54, pp. 1165-1172, 2004.

Taechowisan, T., Peberdy, J.F. & Lumyong, S., Isolation of Endophytic Actinomycetes from Selected Plants and Their Antifungal Activity, World Journal of Microbiology and Biotechnology, 19, pp. 381-385, 2003.

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Published

2021-05-20

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