Bakteri Endofit Penghasil AHL-Laktonase Asal Tanaman Pisang untuk Pengendalian Penyakit Darah
Endophytic Bacteria Producing AHL-Lactonase from Banana Plant to Control Blood Disease
DOI:
https://doi.org/10.14692/jfi.20.6.276-285Keywords:
autoinducer, extracellular polysacharide, quorum sensing, Ralstonia solanacearum, virulence geneAbstract
Endophytic Bacteria Producing AHL-Lactonase from Banana Plant to Control Blood Disease
Blood disease is an important disease in banana that caused 100% yield loss. The Ralstonia solanacearum Phylotype IV as the causal agent for this disease express their virulence genes through quorum sensing (QS) mechanism by using acylated homoserine lactone (AHL) as autoinducer. AHL-lactonase enzyme is produced by bacteria, and its potential as biocontrol agents performed through anti-QS. This research was aimed to obtain endophytic bacteria isolates producing AHL lactonase for controling R. solanacearum in banana plants. The research stages include isolation and selection of endophytic bacteria from banana plant as producer of AHL lactonase by using bioassay and molecular, the effectiveness testing of the bacterial isolates in suppressing R. solanacearum pathogenic virulence factors, in planta testing for controlling R. solanacearum, and molecular identification of potential bacterial isolates. We successfully explored endophytic bacteria from low-, medium-, and up-land consisted several varieties of banana, resulting 152, 204, and 130 isolates, respectively. Further selection using bioindicator showed five isolates with anti quorum sensing activity against Chromobacterium violaceum. Detection of aiiA gene by polymerase chain reaction using specific primer indicated all of five isolates AHL degradation activity resulted 753 bp amplicon that has been identified as aiiA gene. These isolates (CBP1, DBT17, DBT7, DBK2, and CBN13) were able to suppress the formation of exstracellular polysacharide (EPS) produced by R. solanacearum with suppression level of 75.67%, 90.81%, 87.67%, 92.72%, 81.08%, and were able to inhibit wilt symptom in banana plant. Nucleotide sequences of aiiA gene of these five isolates has homology with aiiA genes from Bacillus cereus, B. thuringiensis, B. substilis, B. amyloliquefaciens.
Downloads
References
Amir L. 2015. Isolasi dan karakterisasi bakteri endofit pada tanaman Acasia decurrens (J.C. Wendl.) Willd [tesis]. Bogor (ID): Institut Pertanian Bogor.
Chan KG, Tiew SW, Ng CC. 2007. Rapid isolation method of soil bacilli and screening of their quorum quenching activity. Asia-Pacific Journal of Molecular Biology and Biotechnology. 15(3):153–156.
Chen R, Zhou Z, Cao Y, Bai Y, Yao B. 2010. High yield expression of an AHL-lactonase from Bacillus sp. B546 in Pichia pastoris and its application to reduce Aeromonas hydrophila mortality in aquaculture. Microbial Cell Factories. 9:39. DOI: https://doi.org/10.1186/1475-2859-9-39.
Chong TM, Koh CL, Sam CK, Choo YM, Yin WF, Chan KG. 2012. Characterization of quorum sensing and quorum quenching soil bacteria isolated from Malaysian Tropical Montane Forest. Sensors. 12(4):4846–4859. DOI: https://doi.org/10.3390/s120404846.
Dong YH, Wang LH, Xu JL, Zhang HB, Zhang XF, Zhang LH. 2001. Quenching quorum-sensing-dependent bacterial infection by a N-acyl homoserine lactonase. Nature. 411:813–817. DOI: https://doi.org/10.1038/35081101.
Edy NP, Johanis, Baharuddin. 2008. Karakterisasi morfologi patogenisistas dan biokimia bakteri penyebab penyakit darah pada pisang di Lembah Palu. Jurnal Agrisains 9(2):65–72.
Garg RP, Yindeeyoungyeon W, Gilis A, Denny TP, Van Der LD, Schell MA. 2000. Evidence that Ralstonia eutropha (Alcaligenes eutrophus) contains a functional homologue of the Ralstonia solanacearum Phc cell density sensing system. Molecular microbiology. 38(2):359–367. DOI: https://doi.org/10.1046/j.1365-2958.2000.02131.x.
Gerhardt P, Murray RGE, Wood WA, Noel NR. 1994. Methods for General and Molecular Bacteriology. Washington DC (US): American Society for Microbiology.
Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW. 1997. Bacterial endophytes in agricultural crops. Canadian Journal of Microbiology. 43(10):895–914. DOI: https://doi.org/10.1139/m97-131.
Liu D, Momb J, Thomas PW, Moulin A, Petsko GA, Fast W, Ringe D. 2008. Mechanism of the quorum-quenching lactonase (AiiA) from Bacillus thuringiensis. 1. Product-Bound Structures. Biochemistry. 47(29):
7706–7714. DOI: https://doi.org/10.1021/bi800368y.
Ma A, Lv D, Zhuang X, Zhuang G. 2013. Quorum quenching in culturable phyllosphere bacteria from tobacco. International Journal of Molecular Sciences. 14(7):14607–14619. DOI: https://doi.org/10.3390/ijms140714607.
Marwan H, Sinaga MS, Giyanto, Nawangsih AA. 2011. Isolasi dan seleksi bakteri endofit untuk pengendalian penyakit darah pada tanaman pisang. Jurnal Hama dan Penyakit Tumbuhan Tropika. 11(2):113–121.
McLean RJC, Pierson III LS, Fuqua C. 2004. A simple protocol for the identification of quorum signal antagonists. Journal of Microbiological Methods. 58(3):351–360. DOI: https://doi.org/10.1016/j.mimet.2004.04.016.
Munif A, Wiyono S, Auwarno. 2012. Isolasi bakteri endofit asal padi gogo dan potensinya sebagai agens biokontrol dan pemacu pertumbuhan. Jurnal Fitopatologi Indonesia. 8(3):57–64. DOI: https://doi.org/10.14692/jfi.8.3.57.
Nudyanto A, Zubaidah E. 2015. Isolasi bakteri asam laktat penghasil eksopolisakarida dari kimchi. Jurnal Pangan dan Agroindustri. 3(2):743–748.
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. DOI: https://doi.org/10.1016/j.micres.2007.12.002.
Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular cloning. New York (US): Cold Spring Harbor Laboratory Press.
Sambrook J, Russell D. 2001. Molecular Cloning: A Laboratory Manual. New York (NY): Cold Spring Harbor Laboratory Press.
Schaad NW, Jones JB, Chun W. 2001. Laboratory Guide for Identification of Plant Pathogenic Bacteria. Minnesota (MN): APS Press.
Song C, Ma H, Zhao Q, Song S, Jia Z. 2012. Inhibition of quorum sensing activity by ethanol extract of Scutellaria baicalensis Georgi. Journal of Plant Pathology and Microbiology. 2012(7):1–4. DOI: https://doi.org/10.4172/2157-7471.S7-001.
White CE, Finan TM. 2009. Quorum quenching in Agrobacterium tumefaciens: chance or necessity?. Journal of Bacteriology. 191(4):1123–1125. DOI: https://doi.org/10.1128/JB.01681-08.
Yin XT, Xu L, Fan SS, Xu LN, Li DC, Liu ZY. 2010. Isolation and characterization of an AHL lactonase gene from Bacillus amyloliquefaciens. World Journal of Microbiology Biotechnology. 26(8):1361–1367. DOI: https://doi.org/10.1007/s11274-010-0308-8.
Downloads
Published
Issue
Section
License
Copyright (c) 2025 Jurnal Fitopatologi Indonesia
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Authors who publish in Jurnal Fitopatologi Indonesia agree to the following terms:
1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (CC BY-SA) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository), with an acknowledgement of its initial publication in this journal.
3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.
