Seleksi dan Karakterisasi Bakteri Penghasil Kitinase Penghambat Pertumbuhan Cendawan Patogen Tanaman
Abstract
Colletotrichum capsici and Fusarium oxysporum are pathogenic fungi that cause anthracnose disease on chili plants (Capsicum annum L.) which can decrease the productivity of chili. The control of the fungus attack is carried out by biocontrol agent of the fungi utilizing chitinase-producing bacteria. Chitinase is an enzyme capable of destroying the fungal cell wall composed of chitin compounds. The aims of the research were to select and to characterize chitinase producing bacteria as biological control agent of Colletotrichum capsici and Fusarium oxysporum. Several methods were carried out in this research, among others: isolation of chitinase bacteria, characterization of chitinase bacteria, measurement of enzyme activity, bacterial antagonistic with pathogenic fungi, observation of fungal hypha damage, and identification of 16S rRNA. The BAE36 and BAD211 isolates are chitinase bacteria isolated from chili plant rhizosphere. The isolate belongs to the Gram-negative bacteria. The inhibitory properties of BAE36 and BAD211 against F. oxysporum and C. capsici are on a scale of 3 (51-75%). The percentage of inhibitory of BAE36 isolate was 66.1% capable of inhibiting the growth of C. capsici and 55% capable of inhibiting the growth of F. oxysporum. The hyphae of C. capsici fungus on the antagonistic test showed a thick and the surface of the hyphae was wavy while the F. oxysporum hyphae was thin and looks empty. Identification of isolate with 16S rRNA described that 99% of BAE36 and BAD211 isolates were similar to Lysobacter enzymogenes. Precipitation of chitinase enzyme of Lysobacter enzymogenes with 30% ammonium sulphate could increase the enzyme activity. The enzyme precipitation could inhibit the growth of mycelia fungi C. capsici and F. oxysporum.
Keywords: chili, chitinolytic, Gram-negative, Lysobacter
Downloads
References
Asril M, Mubarik NR, Wahyudi AT. 2014. Partial purification of bacterial chitinase as biocontrol of leaf-blight disease on oil palm. Research Journal of Microbiology. 9(6): 265-277. https://doi.org/10.3923/jm.2014.265.277
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 72(2): 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
Christensen P, Cook FD. 1978. Lysobacter, a new genus of nonfruting, gliding bacteria with a high base ratio. International Journal of Systematic and Bacteriology. 28(3): 367-393. https://doi.org/10.1099/00207713-28-3-367
Dewi RTK, Mubarik NR, Suhartono MT. 2016. Medium optimization of β-glucanase production by Bacillus subtilis SAHA 32.6 used as biological control of oil palm pathogen. Emirates Journal of Food and Agriculture. 28(2): 116-125. https://doi.org/10.9755/ejfa.2015-05-195
Fokkema NJ. 1973. The role of saprophytic fungi in antogonism against Drechslera sorokiniana (Helminthosporium sativum) on agar plats and on rye leaves with pollen. Physiology Plant Phathology. 3: 195-205. https://doi.org/10.1016/0048-4059(73)90082-9
Garg G, Loganathan M, Saha S, Roy BK. 2014. Chilli anthracnose: A review of causal organism, resistence, source and mapping of gene. Microbial Diversity and Biotechnology in Food Security. 4: 589-610. https://doi.org/10.1007/978-81-322-1801-2_53
Joshi M, Srivastava R, Sharma AK, Prakash A. 2012. Screening of resistant varietas and antagonis tic Fusarium oxysporum for biocontrol of Fusarium Wilt of chilli. Journal of Plant Pathology & Microbiology. 3(5): 1-6. https://doi.org/10.4172/2157-7471.1000134
Kambar Y, Manasa M, Viviek MN, Kekuda TRP. 2014. Inhibitory effect of some plants of Western Ghats of Karnataka against Colletotrichum capsici. Science, Technology and Arts Research Journal. 3(2): 76-82. https://doi.org/10.4314/star.v3i2.10
Li S, Jochum CC, Yu F, Zaleta-Rivera K, Du L, Haris SD, Yuen GY. 2008. An antibiotic complex from Lysobacter enzymogenes Strain C3: Antimicrobial activity and role in plant disease control. Phytopahology. 98: 695-701. https://doi.org/10.1094/PHYTO-98-6-069
Luduena LM, Taurian T, Tonelli ML, Angelini JG, Anzuay MS, Valleti L, Munoz V, Fabra AI. 2012. Biocontrol bacterial communities associated with diseased peanut (Arachis hypogaea L.) plants. European Journal of Soil Biology. 53: 48-55. https://doi.org/10.1016/j.ejsobi.2012.08.002
Marchesi JR, Sato T, Weightman AJ, Martin TA, Fry JC, Hiom SJ, Wade WG. 1998. Design and evolution of useful bacteria specific PCR primers that amplify genes coding for bacteria 16S rRNA. Applied and Environmental Microbiology. 64(2): 795-799.
Morris CE, Sands DC, Vinatzer BA, Glaux C, Guilbaund C, Buffierre A, Yan S, Dominguez H, Thompson BM. 2008. The life history of the plant pathogen Pseudomonas syringae is linked to the water cycle. JISME (Multidisciplinary Journal of Microbial Ecology. 2: 321-334. https://doi.org/10.1038/ismej.2007.113
Nurdin GM, Mubarik NR, Sudirman LI. 2016. Selection of chitinolytic bacteria as biological control of Colletotrichum capsici. Malaysian Journal of Microbiology. 12(1): 35-42. https://doi.org/10.21161/mjm.74515
Ozbay N, Newman SE. 2004. Fusarium crown and root rot of tomato and control method. The Plant Pathology Journal. 3(1): 9-18. https://doi.org/10.3923/ppj.2004.9.18
[PDSIP] Pusat Data dan Sistim Informasi Pertanian. 2014. Statistik Iklim, Organisme Pengganggu Tanaman dan Dampak Perubahan Iklim 2011-2013. Jakarta (ID): Sekretariat Jendral, Kementrian Pertanian.
Slimene IB, Tabbene O, Djebali N, Cosette P, Schmitter JM, Jouenne T, Urdaci MC, Limam F. 2012. Puttative use of Bacillus subtilis L194 strain for biocontrol of Phoma medicaginis in Medicago truncatula seedlings. Research in Microbiology. 163: 388-397. https://doi.org/10.1016/j.resmic. 2012.03.004
Soesanto L, Mugiastuti E, Rahayuniati RF. 2010. Kajian mekanisme antagonis Pseudomonas flourescens P60 terhadap Fusarium oxysporum F.SP. lycopersici pada tanaman tomat in vivo. Jurnal Hama dan Penyakit Tumbuhan Tropika. 10(2): 108-115.
Spindler KD. 1997. Chitinase and chitosanase assays. In: Muzarelli RAA, MGPeter, editors. Chitin Handbook. Grottamare (IT): Alda Tecnografica.
Sultana T, Naz F, Haq M, Butt S, Abas MF. 2014. Characterization and relative contribution of fungal and vacterial pathogen involved in sudden death syndrom of chillies. Pakistan Journal of Phytopathology. 26(1): 53-61.
Supriadi. 2006. Analisis risiko agen hayati untuk pengendalian patogen pada tanaman. Journal Penelitian dan Pengembangan Pertanian. 25(3): 75-80.
Tangavelu R, Mustaffa MM. 2012. Current advances in the Fusarium wilt disease management in banana with emphasis on biocontrol, plant phatology. Cumagun DJ, editor. Croatia (HR): In Tech Europe. https://doi.org/10.5772/33775
Wahyudi AT, Maliah S, Nawangsih AA. 2011. Xantomonas oryzae pv. oryzae bakteri penyebab hawar daun pada padi: Isolasi, karakterisasi, dan telaah mutagenesis dengan transposon. Makara sains. 15(1): 89-96.
Zivkovic S, Stojanovic S, Ivanovic Z, Gavrilovic V, Popovic T, Balaz J. 2010. Screening of antagonistic activity of microorganisms against Colletotrichum acutatum and Colletotrichum gloeosporioides. Archives of Biological Sciences. 62(3): 611-623. https://doi.org/10.2298/ABS1003611Z
This journal is published under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License. Authors who publish with this journal agree to the following terms: Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. NonCommercial — You may not use the material for commercial purposes.