The potential of endophytic bacteria from the root of Eucalyptus pellita as a biocontrol agent against Ralstonia solanacearum
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Abstract. Susanti Y, Giyanto, Sinaga MS, Mutaqin KH, Tjahjono B. 2021. The potential of endophytic bacteria from the root of Eucalyptus pellita as a biocontrol agent against Ralstonia solanacearum. Biodiversitas 22: 3454-3462. Bacterial wilt disease (BWD) caused by Ralstonia solanacearum is one of the most damaging eucalyptus (Eucalyptus pellita F. Muell.) diseases. However, endophytic bacteria can be utilized to control it. This study aims to obtain and screen endophytic bacteria isolated from the root of eucalyptus as a biocontrol agent against BWD. A pathogenic R. solanacearum was isolated for artificial inoculation; subsequently, isolation, characterization and screening of the potential endophytic bacteria were conducted. Isolates of R. solanacearum demonstrated the pathogenicity for BWD in eucalyptus, with the highest virulence found in Rs 18. In total, 24 isolates of endophytic bacteria were obtained and screened for their biocontrol potential. The endophytic bacteria conveyed the capacity of antibiosis, indole acetic acid (IAA) hormone, nitrogen fixation, phosphate dissolution, protease enzyme and AHL-lactonase, which are the main indicators of bioagent characteristics in endophytic bacteria, as well as being plant growth stimulators. Four isolates were selected with the highest according to the analytic hierarchy process (AHP), identified on the 16S rRNA gene. Most isolates belonged to gram-negative bacteria. The four isolates, namely B33/77 (Serratia nematodiphila), B32/77 (Paenibacillus polymyxa), B13/497 (Brevundimonas olei) and B17/111 (Bacillus megaterium), indicated their potential for biocontrol as plant growth promoters. This study is the first report on the isolation of endophytic bacterial from eucalyptus roots with regard to its benefit as a plant growth promoter. It provides basic information about the characteristics of the endophytic bacteria, which showed potential as a biocontrol agent of R. solanacearum on eucalyptus.
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REFERENCE
Achari GA & Ramesh R. 2015. Characterization of bacteria degrading 3-hydroxy palmitic acid methyl ester ( 3OH-PAME ), a quorum sensing molecule of Ralstonia solanacearum. Applied Microbiology. 60: 447–455.
Afzal I, Shinwari ZK, Sikandar S, Shahzad S. 2019. Plant beneficial endophytic bacteria: Mechanisms, diversity, host range and genetic determinants. Microbiological Research. 221: 36–49.
Ahemad M. 2014. Phosphate-solubilizing bacteria-assisted phytoremediation of metalliferous soils?: a review. Biotech. 5: 111–121.
Alori ET, Glick BR, Babalola O. 2017. Microbial Phosphorus Solubilization and Its Potential for Use in Sustainable Agriculture. 8(971): 1–8.
Amaresan N, Jayakumar V, Kumar K, Thajuddin N, Nadu T, Islands N, Division CP. 2012. Endophytic bacteria from tomato and chilli, their diversity and antagonistic potential against Ralstonia solanacearum. Archives of Phytopathology and Plant Protection. 45(3): 37–41.
Arwiyanto T. 2014. Ralstonia solanacearum, biologi, penyakit yang ditimbulkan dan pengelolaannya. Gadjah Mada University Press. Yogyakarta.
Badan Pusat Statistik (BPS). 2019. Statistik Perusahaan Pembudidaya Tanaman Kehutanan. Jakarta (ID): Badan Pusat Statistika
Balint-Kurti P. 2019. The plant hypersensitive response?: concepts , control and consequences. Molecular Plant Pathology. 20(8): 1163–1178.
Carstensen GD, Venter SN, Wingfield MJ, Coutinho TA. 2017. Two Ralstonia species associated with bacterial wilt of Eucalyptus. Plant Pathology. 66(3): 393–403.
Coutinho TA & Wingfield MJ. 2017. Ralstonia solanacearum and R. Pseudosolanacearum on eucalyptus: Opportunists or primary pathogens? Frontiers in Plant Science. 8: 1–7.
Doty SL. 2017. Endophytic N-Fixation?: Controversy and a Path Forward. In : Doty SL (eds) Functional Importance of the Plant Microbiome. Springer International Publishing AG. New York
Eyles A, Beadle C, Barry K, Francis A, Glen M, Mohammed C. 2008. Management of fungal root-rot pathogens in tropical Acacia mangium plantations. Forest Pathology. 38(5): 332–355.
Ferreira MA, Mafia RG, Alfenas AC. 2017. Ralstonia solanacearum decreases volumetric growth of trees and yield of kraft cellulose of Eucalyptus spp. Forest Pathology. 48(1): 1–5.
Fonseca NR, Oliveira LSS, Guimarães LMS, Teixeira RU, Lopes CA, Alfenas AC. 2015. An efficient inoculation method of Ralstonia solanacearum to test wilt resistance in Eucalyptus spp. Tropical Plant Pathology. 41(1): 42–47.
Fu S, Wei J, Chen H, Liu Y, Lu H, Chou J. 2015. Indole-3-acetic acid?: A widespread physiological code in interactions of fungi with other organisms. Plant Signaling & Behavior. 10(8): 1–9.
Ghevariya K & Desai P. 2014. Rhizobacteria of sugarcane: In vitro screening for their plant Growth Promoting potentials. Research Journal of Recent Sciences. 3: 52–58.
Glen M, Bougher NL, Francis AA, Nigg SQ, Lee SS, Irianto R, Barry KM, Beadle CL, Mohammed CL. 2009. Ganoderma and Amauroderma species associated with root-rot disease of Acacia mangium plantation trees in Indonesia and Malaysia. Australasian Plant Pathology. 38(4): 345–356.
Guan L, Tayengwa R, Cheng ZM, Peer WA, Murphy AS. 2019. Auxin regulates adventitious root formation in tomato cuttings. BMC Plant Biology. 19(435): 1–16.
Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW. 1997. Bacterial endophytes in agricultural crops. Canadian Journal of Microbiology. 43(10): 895–914.
Hazra F & Susanti WI. 2017. Panduan Praktikum Bioteknologi tanah. Edisi 1. IPB Press.
Hu HQ, Li XS, He H. 2010. Characterization of an antimicrobial material from a newly isolated Bacillus amyloliquefaciens from mangrove for biocontrol of Capsicum bacterial wilt. Biological Control. 54(3): 359–365.
Hung TD, Brawner JT, Meder R, Lee DJ, Southerton S, Thinh HH, Dieters MJ. 2015. Estimates of genetic parameters for growth and wood properties in Eucalyptus pellita F. Muell. to support tree breeding in Vietnam. Annals of Forest Science. 72(2): 205–217.
Islam A, Kabir S, Khair A. 2019. Molecular Identification and Evaluation of Indigenous Bacterial Isolates for Their Plant Growth Promoting and Biological Control Activities against Fusarium Wilt Pathogen of Tomato. Journal Plant Pathol. 35(2): 137–148.
Janda JM & Abbott SL. 2007. 16S rRNA gene sequencing for bacterial identification in the diagnostic laboratory: Pluses, perils, and pitfalls. Journal of Clinical Microbiology. 45(9): 2761–2764.
Jiang H, Dong H, Zhang G, Chapman LR, Fields MW, Jiang H, Dong H, Zhang G, Yu B, Chapman LR, Fields MW. 2006. Microbial Diversity in Water and Sediment of Lake Chaka, an Athalassohaline Lake in Northwestern China. Appl. Environ. Microbiol. 72(6): 3832–2845.
Kandel SL, Joubert PM, Doty SL. 2017. Bacterial Endophyte Colonization and Distribution within Plants. Microorganisms. 5(77): 1–26.
Khan AL, Shahzad R, Al-harrasi A, Lee IJ. 2017. A Resource for Producing Extracellular Enzymes. In : Maheshwari DK and Annapurna K (eds) Endophytes: Crop Productivity and Protection. Springer International Publishing AG. New York
Krzymowska M, Konopka-postupolska D, Sobczak M, Macioszek V, Ellis BE. 2007. Infection of tobacco with different Pseudomonas syringae pathovars leads to distinct morphotypes of programmed cell death. Plant Journal. 50: 253–264.
Kunkel BN & Harper CP. 2018. The roles of auxin during interactions between bacterial plant pathogens and their hosts. 69(2): 245–254.
Lasarre B & Federle MJ. 2013. Exploiting Quorum Sensing To Confuse Bacterial Pathogens. Microbiology and Molecular Biology. 77(1): 73–111.
Liu H, Carvalhais LC, Crawford M, Singh E, Dennis PG, Pieterse CMJ, Schenk PM. 2017. Inner Plant Values?: Diversity, Colonization and Benefits from Endophytic Bacteria. Frontiers in Microbiology. 8(2552): 1–17.
Lyon G. 2007. Agents that can elicit induced resistance. In: Dale Walters, Adrian Newton, and Gary Lyon editor. Induced Resistance for Plant Defence: Sustainable Approach to Crop Protection. Blackwell Publishing
Machmud M 1985 Bacterial wilt in Indonesia Bacterial Wilt Disease in Asia and The South Pacific ed Craswell ET and Pushperajah E (Canberra: ACIAR) pp 30-34
Olatunji D, Geelen D, Verstraeten I. 2017. Control of Endogenous Auxin Levels in Plant Root Development. International Journal of Molecular Sciences. 18(2587): 1–29.
Pan J, Huang T, Yao F, Huang Z, Powell CA, Qiu S, Guan X. 2008. Expression and characterization of aiiA gene from Bacillus subtilis BS-1. Microbiological Research. 163(6): 711–716.
Paz IC, Santin RC, Guimaraes AM, Rosa OP, Dias AC, Quecine MC, Azevedo JL, Matsumura AT. 2012. Eucalyptus growth promotion by endophytic Bacillus spp. Genetics and Molecular Research. 11(4): 3711–3720.
Prasannath K. 2013. Review Article Pathogenicity and Virulence Factors of Phytobacteria. 1(1): 24–33.
Puri A, Padda KP, Chanway CP. 2018. Nitrogen-Fixation by Endophytic Bacteria in Agricultural Crops: Recent Advances. in : Khan A & Fahad S (eds) Nitrogen in Agriculture. InTech. Europa
Ran L, Liu C, Wu G, VanLoon L, Bakker P. 2005. Suppression of bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp. in China. Biological Control. 32(1): 111–120.
Rehman ZU & Leiknes T. 2018. Quorum-Quenching Bacteria Isolated From Red Sea Sediments Reduce Biofilm Formation by Pseudomonas aeruginosa. 9: 1–13.
Rori CA, Ester F, Kandou F, Tangapo AM. 2020. Aktivitas Enzim Ekstraseluler dari Bakteri Endofit Tumbuhan Mangrove Avicennia marina. Jurnal Bios Logios. 10(2): 48–55.
Saaty TL. 2008. Decision making with the analytic hierarchy process. Journal Services Sciences. 1(1): 83–98
Salme timmusk, Copolovici D, Copolovic L, Tede T, Nevo E, Behers L. 2019. Paenibacillus polymyxa biofilm polysaccharides antagonise Fusarium graminearum. Scientific Reports. 9(662): 1–11.
Schaad NW, Jones JB, Chun W. 2001. Initial Identification of Common Genera. In Laboratory Guide for identification of Plant Pathogenic Bacteria. Third edition. 1-35. Edited by Schaad NW, Jones JB, and Chun W. St Paul, MN:APS Press.
Singh M, Kumar A, Singh R, Pandey KD. 2017. Endophytic bacteria: a new source of bioactive compounds. 3 Biotech. 7(5):315–329.
Siregar BA, Indrayadi H dan Mardai 2012. Ancaman Penyakit layu bakteri terhadap produktivitas Eucalyptus pellita di dataran rendah tropis Laporan Tahunan R&D (Perawang: R&D PT. Arara Abadi) pp 15
Soenens A & Imperial J. 2020. Biocontrol capabilities of the genus Serratia. Phytochemistry Reviews. 19(3): 577–587.
Suardana IW, Utama IH, Wibowo H. 2014. Identifikasi escherichia coli o157:h7 dari feses ayam dan uji profil hemolisisnya pada media agar darah. Jurnal Kedokteran Hewan. 8(1): 1–5.
Vasconez RDA, Moya EMT, Yepez LAC, Chiluisa-Utreras VP, Suquillo IAV. 2020. Evaluation of Bacillus megaterium strain AB4 as a potential biocontrol agent of Alternaria japonica, a mycopathogen of Brassica oleracea var. italica. Biotechnology Reports Journal. 26: 1–6.
Velho-Pereira S & Kamat NM. 2011. Antimicrobial screening of actinobacteria using a modified cross-streak method. Indian Journal of Pharmaceutical Sciences. 73(2): 223–228.
Walia A, Guleria S, Chauhan A, Mehta P. 2017. Endophytic Bacteria?: Role in Phosphate Solubilization. in?: Maheshwari DK & Annapurna K(eds), Endophytes : Crop Productivity and Protection. Springer International Publishing AG. New York
Zimbro MJ, Power DA, Miller SM, Wilson GE, Johnson JA. 2009. Difco & BBL Manual: Manual of Microbiological Culture Media. Becton. Dickinson and Company. USA
Achari GA & Ramesh R. 2015. Characterization of bacteria degrading 3-hydroxy palmitic acid methyl ester ( 3OH-PAME ), a quorum sensing molecule of Ralstonia solanacearum. Applied Microbiology. 60: 447–455.
Afzal I, Shinwari ZK, Sikandar S, Shahzad S. 2019. Plant beneficial endophytic bacteria: Mechanisms, diversity, host range and genetic determinants. Microbiological Research. 221: 36–49.
Ahemad M. 2014. Phosphate-solubilizing bacteria-assisted phytoremediation of metalliferous soils?: a review. Biotech. 5: 111–121.
Alori ET, Glick BR, Babalola O. 2017. Microbial Phosphorus Solubilization and Its Potential for Use in Sustainable Agriculture. 8(971): 1–8.
Amaresan N, Jayakumar V, Kumar K, Thajuddin N, Nadu T, Islands N, Division CP. 2012. Endophytic bacteria from tomato and chilli, their diversity and antagonistic potential against Ralstonia solanacearum. Archives of Phytopathology and Plant Protection. 45(3): 37–41.
Arwiyanto T. 2014. Ralstonia solanacearum, biologi, penyakit yang ditimbulkan dan pengelolaannya. Gadjah Mada University Press. Yogyakarta.
Badan Pusat Statistik (BPS). 2019. Statistik Perusahaan Pembudidaya Tanaman Kehutanan. Jakarta (ID): Badan Pusat Statistika
Balint-Kurti P. 2019. The plant hypersensitive response?: concepts , control and consequences. Molecular Plant Pathology. 20(8): 1163–1178.
Carstensen GD, Venter SN, Wingfield MJ, Coutinho TA. 2017. Two Ralstonia species associated with bacterial wilt of Eucalyptus. Plant Pathology. 66(3): 393–403.
Coutinho TA & Wingfield MJ. 2017. Ralstonia solanacearum and R. Pseudosolanacearum on eucalyptus: Opportunists or primary pathogens? Frontiers in Plant Science. 8: 1–7.
Doty SL. 2017. Endophytic N-Fixation?: Controversy and a Path Forward. In : Doty SL (eds) Functional Importance of the Plant Microbiome. Springer International Publishing AG. New York
Eyles A, Beadle C, Barry K, Francis A, Glen M, Mohammed C. 2008. Management of fungal root-rot pathogens in tropical Acacia mangium plantations. Forest Pathology. 38(5): 332–355.
Ferreira MA, Mafia RG, Alfenas AC. 2017. Ralstonia solanacearum decreases volumetric growth of trees and yield of kraft cellulose of Eucalyptus spp. Forest Pathology. 48(1): 1–5.
Fonseca NR, Oliveira LSS, Guimarães LMS, Teixeira RU, Lopes CA, Alfenas AC. 2015. An efficient inoculation method of Ralstonia solanacearum to test wilt resistance in Eucalyptus spp. Tropical Plant Pathology. 41(1): 42–47.
Fu S, Wei J, Chen H, Liu Y, Lu H, Chou J. 2015. Indole-3-acetic acid?: A widespread physiological code in interactions of fungi with other organisms. Plant Signaling & Behavior. 10(8): 1–9.
Ghevariya K & Desai P. 2014. Rhizobacteria of sugarcane: In vitro screening for their plant Growth Promoting potentials. Research Journal of Recent Sciences. 3: 52–58.
Glen M, Bougher NL, Francis AA, Nigg SQ, Lee SS, Irianto R, Barry KM, Beadle CL, Mohammed CL. 2009. Ganoderma and Amauroderma species associated with root-rot disease of Acacia mangium plantation trees in Indonesia and Malaysia. Australasian Plant Pathology. 38(4): 345–356.
Guan L, Tayengwa R, Cheng ZM, Peer WA, Murphy AS. 2019. Auxin regulates adventitious root formation in tomato cuttings. BMC Plant Biology. 19(435): 1–16.
Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW. 1997. Bacterial endophytes in agricultural crops. Canadian Journal of Microbiology. 43(10): 895–914.
Hazra F & Susanti WI. 2017. Panduan Praktikum Bioteknologi tanah. Edisi 1. IPB Press.
Hu HQ, Li XS, He H. 2010. Characterization of an antimicrobial material from a newly isolated Bacillus amyloliquefaciens from mangrove for biocontrol of Capsicum bacterial wilt. Biological Control. 54(3): 359–365.
Hung TD, Brawner JT, Meder R, Lee DJ, Southerton S, Thinh HH, Dieters MJ. 2015. Estimates of genetic parameters for growth and wood properties in Eucalyptus pellita F. Muell. to support tree breeding in Vietnam. Annals of Forest Science. 72(2): 205–217.
Islam A, Kabir S, Khair A. 2019. Molecular Identification and Evaluation of Indigenous Bacterial Isolates for Their Plant Growth Promoting and Biological Control Activities against Fusarium Wilt Pathogen of Tomato. Journal Plant Pathol. 35(2): 137–148.
Janda JM & Abbott SL. 2007. 16S rRNA gene sequencing for bacterial identification in the diagnostic laboratory: Pluses, perils, and pitfalls. Journal of Clinical Microbiology. 45(9): 2761–2764.
Jiang H, Dong H, Zhang G, Chapman LR, Fields MW, Jiang H, Dong H, Zhang G, Yu B, Chapman LR, Fields MW. 2006. Microbial Diversity in Water and Sediment of Lake Chaka, an Athalassohaline Lake in Northwestern China. Appl. Environ. Microbiol. 72(6): 3832–2845.
Kandel SL, Joubert PM, Doty SL. 2017. Bacterial Endophyte Colonization and Distribution within Plants. Microorganisms. 5(77): 1–26.
Khan AL, Shahzad R, Al-harrasi A, Lee IJ. 2017. A Resource for Producing Extracellular Enzymes. In : Maheshwari DK and Annapurna K (eds) Endophytes: Crop Productivity and Protection. Springer International Publishing AG. New York
Krzymowska M, Konopka-postupolska D, Sobczak M, Macioszek V, Ellis BE. 2007. Infection of tobacco with different Pseudomonas syringae pathovars leads to distinct morphotypes of programmed cell death. Plant Journal. 50: 253–264.
Kunkel BN & Harper CP. 2018. The roles of auxin during interactions between bacterial plant pathogens and their hosts. 69(2): 245–254.
Lasarre B & Federle MJ. 2013. Exploiting Quorum Sensing To Confuse Bacterial Pathogens. Microbiology and Molecular Biology. 77(1): 73–111.
Liu H, Carvalhais LC, Crawford M, Singh E, Dennis PG, Pieterse CMJ, Schenk PM. 2017. Inner Plant Values?: Diversity, Colonization and Benefits from Endophytic Bacteria. Frontiers in Microbiology. 8(2552): 1–17.
Lyon G. 2007. Agents that can elicit induced resistance. In: Dale Walters, Adrian Newton, and Gary Lyon editor. Induced Resistance for Plant Defence: Sustainable Approach to Crop Protection. Blackwell Publishing
Machmud M 1985 Bacterial wilt in Indonesia Bacterial Wilt Disease in Asia and The South Pacific ed Craswell ET and Pushperajah E (Canberra: ACIAR) pp 30-34
Olatunji D, Geelen D, Verstraeten I. 2017. Control of Endogenous Auxin Levels in Plant Root Development. International Journal of Molecular Sciences. 18(2587): 1–29.
Pan J, Huang T, Yao F, Huang Z, Powell CA, Qiu S, Guan X. 2008. Expression and characterization of aiiA gene from Bacillus subtilis BS-1. Microbiological Research. 163(6): 711–716.
Paz IC, Santin RC, Guimaraes AM, Rosa OP, Dias AC, Quecine MC, Azevedo JL, Matsumura AT. 2012. Eucalyptus growth promotion by endophytic Bacillus spp. Genetics and Molecular Research. 11(4): 3711–3720.
Prasannath K. 2013. Review Article Pathogenicity and Virulence Factors of Phytobacteria. 1(1): 24–33.
Puri A, Padda KP, Chanway CP. 2018. Nitrogen-Fixation by Endophytic Bacteria in Agricultural Crops: Recent Advances. in : Khan A & Fahad S (eds) Nitrogen in Agriculture. InTech. Europa
Ran L, Liu C, Wu G, VanLoon L, Bakker P. 2005. Suppression of bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp. in China. Biological Control. 32(1): 111–120.
Rehman ZU & Leiknes T. 2018. Quorum-Quenching Bacteria Isolated From Red Sea Sediments Reduce Biofilm Formation by Pseudomonas aeruginosa. 9: 1–13.
Rori CA, Ester F, Kandou F, Tangapo AM. 2020. Aktivitas Enzim Ekstraseluler dari Bakteri Endofit Tumbuhan Mangrove Avicennia marina. Jurnal Bios Logios. 10(2): 48–55.
Saaty TL. 2008. Decision making with the analytic hierarchy process. Journal Services Sciences. 1(1): 83–98
Salme timmusk, Copolovici D, Copolovic L, Tede T, Nevo E, Behers L. 2019. Paenibacillus polymyxa biofilm polysaccharides antagonise Fusarium graminearum. Scientific Reports. 9(662): 1–11.
Schaad NW, Jones JB, Chun W. 2001. Initial Identification of Common Genera. In Laboratory Guide for identification of Plant Pathogenic Bacteria. Third edition. 1-35. Edited by Schaad NW, Jones JB, and Chun W. St Paul, MN:APS Press.
Singh M, Kumar A, Singh R, Pandey KD. 2017. Endophytic bacteria: a new source of bioactive compounds. 3 Biotech. 7(5):315–329.
Siregar BA, Indrayadi H dan Mardai 2012. Ancaman Penyakit layu bakteri terhadap produktivitas Eucalyptus pellita di dataran rendah tropis Laporan Tahunan R&D (Perawang: R&D PT. Arara Abadi) pp 15
Soenens A & Imperial J. 2020. Biocontrol capabilities of the genus Serratia. Phytochemistry Reviews. 19(3): 577–587.
Suardana IW, Utama IH, Wibowo H. 2014. Identifikasi escherichia coli o157:h7 dari feses ayam dan uji profil hemolisisnya pada media agar darah. Jurnal Kedokteran Hewan. 8(1): 1–5.
Vasconez RDA, Moya EMT, Yepez LAC, Chiluisa-Utreras VP, Suquillo IAV. 2020. Evaluation of Bacillus megaterium strain AB4 as a potential biocontrol agent of Alternaria japonica, a mycopathogen of Brassica oleracea var. italica. Biotechnology Reports Journal. 26: 1–6.
Velho-Pereira S & Kamat NM. 2011. Antimicrobial screening of actinobacteria using a modified cross-streak method. Indian Journal of Pharmaceutical Sciences. 73(2): 223–228.
Walia A, Guleria S, Chauhan A, Mehta P. 2017. Endophytic Bacteria?: Role in Phosphate Solubilization. in?: Maheshwari DK & Annapurna K(eds), Endophytes : Crop Productivity and Protection. Springer International Publishing AG. New York
Zimbro MJ, Power DA, Miller SM, Wilson GE, Johnson JA. 2009. Difco & BBL Manual: Manual of Microbiological Culture Media. Becton. Dickinson and Company. USA
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