Characterization of Pantoea ananatis Isolated from Garlic and Shallot

https://doi.org/10.22146/jpti.27407

Nanik Nurjanah(1*), Tri Joko(2), Siti Subandiyah(3)

(1) Agricultural Quarantine Agency Class 1 Ambon Jln. Rurehe, Waihaong, Ambon, Maluku 97111
(2) Department of Crop Protection, Faculty of Agriculture, Universitas Gadjah Mada Jln. Flora 1, Bulaksumur, Sleman, Yogyakarta 55281
(3) Department of Crop Protection, Faculty of Agriculture, Universitas Gadjah Mada Jln. Flora 1, Bulaksumur, Sleman, Yogyakarta 55281
(*) Corresponding Author

Abstract


The new disease on garlic (Allium sativum) and shallot (A. cepa L. aggregatum group) have been found in several production centers of garlic and shallot in Tawangmangu and Temanggung, Central Java. The infected plants showed symptoms of leaf blight accompanied by chlorosis. The objective of this study was to determine the pathogen that causes leaf blight and chlorosis based on the phenotypic characterization and gyrB gene sequences analysis. The research started from the isolation of pathogen, physiological and biochemical test, DNA extraction, and sequence analysis of gyrB using gyrB 01-F and gyrB 02-R primer. The results showed that the isolated bacterial pathogen have a yellow pigment, slimy colonies with regular borders, convex, gram-negative, non-spore, facultative anaerobic, motile, catalase production, indole production, and acid production from D-glucose, D-mannitol, sucrose, and lactose. From the pathogenicity test, it was found that the bacteria produced the typical symptom of leaf blight. Characterization of pathogens based on gyrB gene sequence revealed that the pathogen was placed in the group of Pantoea ananatis.

 

Intisari


Penyakit baru pada bawang putih (Allium sativum) dan bawang merah (A. cepa L. aggregatum group) telah ditemukan di beberapa sentra produksi bawang putih dan bawang merah di Tawangmangu dan Temanggung, Jawa Tengah. Tanaman yang terinfeksi menunjukkan gejala hawar daun disertai klorosis. Tujuan penelitian untuk mengetahui karakter patogen berdasarkan fenotipik dan sekuen gen gyrB. Penelitian dimulai dengan isolasi bagian tanaman yang sakit, uji fisiologi dan biokimia, ekstraksi DNA dengan metode CTAB/NaCl dan amplifikasi gen gyrB menggunakan primer gyrB 01-F and gyrB 02-R. Hasil uji menunjukkan koloni berlendir, cembung, pigmen berwarna kuning, gram negative, tidak berspora, aerob fakultatif, motil, produksi katalase, indol, membentuk asam dari D-glukosa, D-monnitol, sukrosa dan laktosa, dan patogenesitas positif. Karakterisasi patogen berdasarkan sekuen gen gyrB, menunjukkan patogen hawar daun berkerabat dekat dengan Pantoea ananatis.


Keywords


Alium cepa; Alium sativum; gyrB; leaf blight; Pantoea ananatis

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References

Aneja, K.R. 2003. Experiments in Microbiology, Plant Pathology and Biotechnology. Fourth Edition. New Age International Publishers, New Delhi. 607 p.

Azad, H.R., G.J. Holmes, & D.A. Cooksey. 2000. A New Leaf Blotch Disease of Sudan Grass Caused by Pantoea ananas and Pantoea stewartii. Plant Disease 84: 973–979.

Carr, E.A., J.M. Bonasera, A.M. Zaid, J.W. Lorbeer, & S.V. Beer. 2010. First Report of Bulb Disease of Onion Caused by Pantoea ananatis in New York. Plant Disease 94: 916.

Choi, O., J.Y. Lim, Y.S. Seo, I. Hwang, & J. Kim. 2012. Complete Genome Sequence of the Rice Pathogen Pantoea ananatis Strain PA13. Journal of Bacteriology 194: 531.

Conn, K., J. Lutton, & S. Rosenberger. 2012. Onion Disease Guide. Seminis Vegetable Seeds, California, USA. 72 p.

Danaatmadja, Y., S. Subandiyah, T. Joko, & C.U. Sari. 2009. Isolasi dan Karakterisasi Ralstonia syzygii. Jurnal Perlindungan Tanaman Indonesia 15: 7−12.

Dwimartina, F., T. Arwiyanto, & T. Joko. 2017. Potential of Endophytic and Rhizobacteria as an Effective Biocontrol for Ralstonia syzygii subsp. syzygii. Asian Journal of Plant Pathology 11: 191–198.

Egorova, M., E. Mazurin, & A.N. Ignatov. 2015. First Report of Pantoea ananatis Causing Grain Discolouration and Leaf Blight of Rice in Russia. New Disease Reports 32: 21.

Gitaitis, R.D. & J.D. Gay. 1997. First Report of Leaf Blight, Seed Stalk Rot, and Bulb Decay of Onion by P. ananatis in Georgia. Plant Disease 81: 1096.

Gitaitis, R.D., R.R. Walcott, M.L. Wells, J.C. Diaz Perez, & F.H. Sanders. 2003. Transmission of P. ananatis, Causal Agent of Center Rot of Onion, by Tobacco Thrips, Frankliniella fusca. Plant Disease 87: 675–678.

Harayama, S. & H. Kasai. 2006. Bacterial Phylogeny Reconstruction from Molecular Sequences. In E. Stackebrandt (ed.) Molecular Identification, Systematics, and Population Structure of Prokaryotes, Chap. 5. Springer, Berlin.

Ismiyatuningsih, T. Joko, & S. Hartono. 2016. Survey and Detection of Pectobacterium atrosepticum in Major Potato-Growing Areas in Central Java Province, Indonesia. Ilmu Pertanian 1: 1−6.

Joko, T., S. Subandiyah, & S. Somowiyarjo. 2000. The Role of Extracellular Protein on the Pathogenicity of Xanthomonas campestris pv. citri. Jurnal Perlindungan Tanaman Indonesia 6: 32−38.

Joko, T., H. Hirata, & S. Tsuyumu. 2007a. Sugar Transporter (MfsX) of Major Facilitator Superfamily is Required for Flagella-Mediated Pathogenesis in Dickeya dadantii 3937. Journal of General Plant Patholology 73: 266−273.

Joko, T., H. Hirata, & S. Tsuyumu, 2007b. A Sugar Transporter (MfsX) is also Required by Dickeya dadantii 3937 for in Planta Fitness. Journal of General Plant Patholology 73: 274−80.

Joko, T., D. Kiswanti, S. Subandiyah, & Hanudin. 2011a. Occurence of Bacterial Soft Rot of Phalaenopsis Orchids in Yogyakarta and West Java, Indonesia, pp. 255–265. In Y. Koentjoro (ed.), Proceeding of Internasional Seminar on “Natural Resources, Climate Change, and Food Security in Developing Countries”, 27−28 June 2011. Surabaya, Indonesia.

Joko, T., N. Kusumandari, & S. Hartono. 2011b. Optimasi Metode PCR untuk Deteksi Pectobacterium carotovorum, Penyebab Penyakit Busuk Lunak Anggrek. Jurnal Perlindungan Tanaman Indonesia 17: 54–59.

Joko, T., M.P. Koentjoro, S. Somowiyarjo, M.S. Rohman, A. Liana, & N. Ogawa. 2012. Response of Rhizobacterial Communities in Watermelon to Infection with Cucumber Green Mottle Mosaic Virus as Revealed by Cultivation-Dependent RISA. Archives of Phytopathology and Plant Protection 45: 1810−1818.

Joko, T., A. Subandi, N. Kusumandari, A. Wibowo, & A. Priyatmojo. 2014. Activities of Plant Cell Wall-Degrading Enzymes by Bacterial Soft Rot of Orchid. Archives of Phytopathology and Plant Protection 47: 1239−1250.

Kido, K., M. Hasegawa, H. Matsumoto, M., Kobayashi, & Y. Takikawa. 2010. Pantoea ananatis Strains are Differentiated Into Three Groups Based on Reactions of Tobacco and Welsh Onion and on Genetic Characteristics. Journal of General Plant Pathology 76: 208–218.

Kumar, S., G. Stecher, & K. Tamura. 2016. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Biology and Evolution 33: 1870–1874.

Leboffe, M.J. & B.E. Pierce. 2010. Microbiology Laboratory Theory and Application, 3rd ed. Morton Publishing Company, Englewood, Co. 656 p.

Lelliot, R.A. & D.E. Stead. 1987. Methods in Plant Pathology. Vol. 2. Methods for the Diagnosis of Bacterial Diseases of Plants. British Society for Plant Pathology by Blackwell Scientific Publication, Oxford. 216 p.

Mahfut, T. Joko, B.S. Daryono, & S. Somowiyarjo. 2016a. Survei Odontoglossum Ringspot Virus (ORSV) yang menginfeksi anggrek alam tropis di Indonesia. Jurnal Perlindungan Tanaman Indonesia 17: 54–59.

Mahfut, T. Joko, & B.S. Daryono. 2016b. Molecular Characterization of Odontoglossum Ringspot Virus (ORSV) in Java and Bali, Indonesia. Asian Journal of Plant Pathology 10: 9−14.

Miller, A.M., J.E.F. Figueiredo, C.L. Chaves, E.A. Ruas, M.I. Balbi-Peña, N.B. Colauto, & L.D. Paccola-Meirelles. 2016. Genomic Variability of P. ananatis in Maize white Spot Lesions Assessed by AFLP Markers. Genetics and Molecular Research 15: 1–13.

Parkinson, N., C. Cowie, J. Heeney, & D. Stead. 2009. Phylogenetic Structure of Xanthomonas Determined by Comparison of gyrB Sequences. International Journal of Systematic and Evolutionary Microbiology 59: 264– 274.

Pérez-y-Terrón, R., M.C. Villegas, A. Cuellar, J. Muñoz-Rojas, M. Castañeda-Lucio, I. Hernández-Lucas, R. Bustillos-Cristales, C.L. Bautista-Sosa, J.A. Munive, R. Caicedo-Rivas, & L.E. Fuentes-Ramírez. 2009. Detection of P. ananatis, Causal Agent of Leaf Spot Disease of Maize, in Mexico. Australasian Plant Disease Notes 4: 96–99.

Rezzonico, F., T.H. Smits, E. Montesinos, J.E. Frey, & B. Duffy. 2009. Genotypic Comparison of Pantoea agglomerans Plant and Clinical Strains. BMC Microbiology 9: 204.

Sauer, A.V., K.R. Rocha, R.M. Gonçalves, & W.F. Meirelles. 2015. Survival of Pantoea ananatis, Causal Agent of Maize White Spot Disease in Crop Debris. Agronomy Science and Biotechnology 1: 21–24.

Schaad, N.W., J.B. Jones, & W. Chun. 2001. Laboratory Guide for Idetification of Plant Pathogenic Bacteria. Third Edition. Minnesota: The American Phytopathological Society. 373 p.

Suharti, T., T. Joko, & T. Arwiyanto. 2017. Deteksi Bakteri Patogen Terbawa Benih Akor (Acacia auriculiformis A. Cunn. ex Benth.). Jurnal Hama dan Penyakit Tumbuhan Tropika 17: 19–36.

Sumardiyono, C., T. Joko, Y. Kristiawati, & Y.D. Chinta. 2011. Diagnosis dan Pengendalian Penyakit Antraknosa pada Pakis dengan Fungisida. Jurnal Hama dan Penyakit Tumbuhan Tropika 11: 194–200.

Takeda, K., Y.Q. Kang, K. Yazawa, T. Gonoi, & Y. Mikami. 2010. Phylogenetic Studies of Nocardia Species Based on gyrB Gene Analyses. Journal of Medical Microbiology 59: 165–171.

Walcott, R.R, R.D. Gitaitis, A.C. Castro, F.H. Sanders, & J.C. Diaz-Perez. 2002. Natural Infestation of Onion Seed by P. ananatis, Causal Agent of Center Rot. Plant Disease 86: 106–111.

Wardhika, C.M., Suryanti, & T. Joko. 2014. Eksplorasi Bakteri Agens Pengendali Hayati Fusarium solani dan Meloidogyne incognita pada Lada. Jurnal Perlindungan Tanaman Indonesia 18: 90−95.

Wibowo, A., T. Joko, S. Subandiyah, I. Mariska, Y. Supriyati, Y. Suryadi, & I. Roostika. 2010. Peningkatan Ketahanan Tanaman Pisang Kepok Kuning Terhadap Penyakit Darah Melalui Variasi Somaklonal dan Simbiosis Endofitik. Jurnal Perlindungan Tanaman Indonesia 16: 15–21.

Widyaningsih, S., S.N.H. Utami, T. Joko, & S. Subandiyah. 2017. Development of Disease and Growth on Six Scion/Rootstock Combinations of Citrus Seedlings under Huanglongbing Pressure. Journal of Agricultural Science 9: 229−238.

Windari, U., T. Joko, & S. Subandiyah. 2015. Deteksi Penyakit Bacterial Fruit Blotch pada Melon Menggunakan Elisa. Jurnal Perlindungan Tanaman Indonesia 19: 1–5.



DOI: https://doi.org/10.22146/jpti.27407

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