Genetic Relationships of Soft Rot Bacteria Isolated from Konjac in China by Amplified Fragment Length Polymorphism (AFLP) and 16S rDNA Gene Sequences
Affiliation(s)
1 Hubei Academy of Agricultural Sciences, Wuhan, China. 2 Enshi Autonomous Prefecture Academy of Agricultural Sciences, Enshi, China. 3 College of Life Sciences, Wuhan University, Wuhan, China.
1 Hubei Academy of Agricultural Sciences, Wuhan, China. 2 Enshi Autonomous Prefecture Academy of Agricultural Sciences, Enshi, China. 3 College of Life Sciences, Wuhan University, Wuhan, China.
ABSTRACT
Twenty-three isolates of soft rot bacteria from konjac corms were examined for their diversity using 16S rDNAs and AFLP technology. Both methods clustered two groups, dependent on their biotype characterization of Pectobacterium carotovora subsp. carotovora (P.c.c) and Pectobacterium chrysanthemi (P.ch), respectively. Of all isolates, 17 (73.9%) belonged to P. ch, indicated as the main pathogenic bacteria of konjac producing areas in China. The genetic variation among isolates from the same biotype was also rich, not consistent with the distances of the geographic sources.
Twenty-three isolates of soft rot bacteria from konjac corms were examined for their diversity using 16S rDNAs and AFLP technology. Both methods clustered two groups, dependent on their biotype characterization of Pectobacterium carotovora subsp. carotovora (P.c.c) and Pectobacterium chrysanthemi (P.ch), respectively. Of all isolates, 17 (73.9%) belonged to P. ch, indicated as the main pathogenic bacteria of konjac producing areas in China. The genetic variation among isolates from the same biotype was also rich, not consistent with the distances of the geographic sources.
Cite this paper
Wu, J. , Yang, C. , Jiao, Z. , Hu, Z. , Ding, Z. and Qiu, Z. (2015) Genetic Relationships of Soft Rot Bacteria Isolated from Konjac in China by Amplified Fragment Length Polymorphism (AFLP) and 16S rDNA Gene Sequences. Agricultural Sciences, 6, 717-723. doi: 10.4236/as.2015.67069.
Wu, J. , Yang, C. , Jiao, Z. , Hu, Z. , Ding, Z. and Qiu, Z. (2015) Genetic Relationships of Soft Rot Bacteria Isolated from Konjac in China by Amplified Fragment Length Polymorphism (AFLP) and 16S rDNA Gene Sequences. Agricultural Sciences, 6, 717-723. doi: 10.4236/as.2015.67069.
References
[1] Vasques, C.A., et al. (2008) Evaluation of the Pharmacotherapeutic Efficacy of Garcinia cambogia plus Amorphophallus konjac for the Treatment of Obesity. Phytotherapy Research, 22, 1135-1140.
http://dx.doi.org/10.1002/ptr.2323 [2] Wu, J.P., et al. (2011) Molecular Detection of Pectobacterium Species Causing Soft Rot of Amorphophallus konjac. World Journal of Microbiology & Biotechnology, 27, 613-618.
http://dx.doi.org/10.1007/s11274-010-0496-2 [3] Gracia-Garza, J.A., et al. (2004) Increased Incidence of Erwinia Soft-Rot on Calla Lilies in the Presence of Phosphorous. European Journal of Plant Pathology, 110, 293-298.
http://dx.doi.org/10.1023/B:EJPP.0000019799.17513.86 [4] Ali, H.F., et al. (2012) Inoculum Sources, Disease Incidence and Severity of Bacterial Blackleg and Soft Rot of Potato. Pakistan Journal of Botany, 44, 825-830. [5] Ngadze, E., et al. (2012) Pectinolytic Bacteria Associated with Potato Soft Rot and Blackleg in South Africa and Zimbabwe. European Journal of Plant Pathology, 134, 533-549.
http://dx.doi.org/10.1007/s10658-012-0036-z [6] Waldee, E.L. (1945) Comparative Studies of Some Peritrichous Phytopathogenic Bacteria. Iowa State College Journal of Science, 19, 435-484. [7] Toth, I., Avrova, A. and Hyman, L. (2001) Rapid Identification and Differentiation of the Soft Rot Erwinias by 16S-23S Intergenic Transcribed Spacer-PCR and Restriction Fragment Length Polymorphism Analyses. Applied and Environmental Microbiology, 67, 4070-4076.
http://dx.doi.org/10.1128/AEM.67.9.4070-4076.2001 [8] Brenner, D.J., Fanning, G.R. and Steigerwalt, A.G. (1977) Deoxyribonucleic Acid Relatedness among Erwiniae and Other Enterobacteria II. Corn Stalk Rot Bacterium and Pectobacterium chrysanthemi. International Journal of Systematic and Evolutionary Microbiology, 27, 211-221.
http://dx.doi.org/10.1099/00207713-27-3-211 [9] Gallois, A., et al. (1992) Erwinia carotovora subsp. odorifera subsp. nov., Associated with Odorous Soft Rot of Chicory (Cichorium intybus L.). International Journal of Systematic and Evolutionary Microbiology, 42, 582-588.
http://dx.doi.org/10.1099/00207713-42-4-582 [10] Nassar, A., et al. (1994) Ribotyping of Erwinia chrysanthemi Strains in Relation to Their Pathogenic and Geographic Distribution. Applied and Environmental Microbiology, 60, 3781-3789. [11] Xiu, J.H., Ji, G.H., Wang, M., Yang, Y.L. and Li, C.Y. (2006) Molecular Identification and Genetic Diversity in Konnyaku’s Soft Rot Bacteria. Acta Microbiologica Sinica, 46, 522-525. [12] Hauben, L., Moore, E.R., Vauterin, L., Steenackers, M., Mergaert, J., Verdonck, L. and Swings, J. (1998) Phylogenetic Position of Phytopathogens within the Enterobacteriaceae. Systematic and Applied Microbiology, 21, 384-397.
http://dx.doi.org/10.1016/S0723-2020(98)80048-9 [13] Kwon, S.W., Go, S.J., Kang, H.W., Ryu, J.C. and Jo, J.K. (1997) Phylogenetic Analysis of Erwinia Species Based on 16S rRNA Gene Sequences. International Journal of Systematic Bacteriology, 47, 1061-1067.
http://dx.doi.org/10.1099/00207713-47-4-1061 [14] Dickey, R.S. (1979) Erwinia chrysanthemi: A Comparative Study of Phenotypic Properties of Strains from Several Hosts and Other Erwinia Species. Phytopathology, 69, 324-329.
http://dx.doi.org/10.1094/Phyto-69-324 [15] Dye, D.W. (1981) A Numerical Taxonomic Study of the Genus Erwinia. New Zealand Journal of Agricultural Research, 24, 223-229.
http://dx.doi.org/10.1080/00288233.1981.10420894 [16] Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes, M., et al. (1995) AFLP: A New Technique for DNA Fingerprinting. Nucleic Acids Research, 23, 4407-4414.
http://dx.doi.org/10.1093/nar/23.21.4407 [17] Geornaras, I., Kunene, N.F., von Holy, A. and Hastings, J.W. (1999) Amplified Fragment Length Polymorphism Fingerprinting of Pseudomonas Strains from a Poultry Processing Plant. Applied and Environmental Microbiology, 65, 3828-3833. [18] Janssen, P., Coopman, R., Huys, G., Swings, J., Bleeker, M., Vos, P., et al. (1996) Evaluation of the DNA Fingerprinting Method AFLP as an New Tool in Bacterial Taxonomy. Microbiology, 142, 1881-1893.
http://dx.doi.org/10.1099/13500872-142-7-1881 [19] Weisburg, W.G., Barns, S.M., Pelletier, D.A. and Lane, D.J. (1991) 16S Ribosomal DNA Amplification for Phylogenetic Study. Journal of Bacteriology, 173, 697-703. [20] Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor. [21] Tamura, K., Stecher, G., Peterson, D., Filipski, A. and Kumar, S. (2013) MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30, 2725-2729.
http://dx.doi.org/10.1093/molbev/mst197 [22] Han, Y.C., Teng, C.Z., Hu, Z.L. and Song, Y.C. (2008) An Optimal Method of DNA Silver Staining in Polyacrylamide Gels. Electrophoresis, 29, 1355-1358.
http://dx.doi.org/10.1002/elps.200700558 [23] Rohlf, F. (2000) NTSYS-pc Version 2.10 m, Numerical Taxonomy and Multivariate Analysis System (Computer Program). Exeter Software, Steauket. [24] Snijder, R.C., Cho, H.-R., Hendriks, M.M.W.B., Lindhout, P. and van Tuyl, J.M. (2004) Genetic Variation in Zantedeschia spp. (Araceae) for Resistance to Soft Rot Caused by Erwinia carotovora subsp. carotovora. Euphytica, 135, 119-128.
http://dx.doi.org/10.1023/B:EUPH.0000009546.88984.60
[1] Vasques, C.A., et al. (2008) Evaluation of the Pharmacotherapeutic Efficacy of Garcinia cambogia plus Amorphophallus konjac for the Treatment of Obesity. Phytotherapy Research, 22, 1135-1140.
http://dx.doi.org/10.1002/ptr.2323 [2] Wu, J.P., et al. (2011) Molecular Detection of Pectobacterium Species Causing Soft Rot of Amorphophallus konjac. World Journal of Microbiology & Biotechnology, 27, 613-618.
http://dx.doi.org/10.1007/s11274-010-0496-2 [3] Gracia-Garza, J.A., et al. (2004) Increased Incidence of Erwinia Soft-Rot on Calla Lilies in the Presence of Phosphorous. European Journal of Plant Pathology, 110, 293-298.
http://dx.doi.org/10.1023/B:EJPP.0000019799.17513.86 [4] Ali, H.F., et al. (2012) Inoculum Sources, Disease Incidence and Severity of Bacterial Blackleg and Soft Rot of Potato. Pakistan Journal of Botany, 44, 825-830. [5] Ngadze, E., et al. (2012) Pectinolytic Bacteria Associated with Potato Soft Rot and Blackleg in South Africa and Zimbabwe. European Journal of Plant Pathology, 134, 533-549.
http://dx.doi.org/10.1007/s10658-012-0036-z [6] Waldee, E.L. (1945) Comparative Studies of Some Peritrichous Phytopathogenic Bacteria. Iowa State College Journal of Science, 19, 435-484. [7] Toth, I., Avrova, A. and Hyman, L. (2001) Rapid Identification and Differentiation of the Soft Rot Erwinias by 16S-23S Intergenic Transcribed Spacer-PCR and Restriction Fragment Length Polymorphism Analyses. Applied and Environmental Microbiology, 67, 4070-4076.
http://dx.doi.org/10.1128/AEM.67.9.4070-4076.2001 [8] Brenner, D.J., Fanning, G.R. and Steigerwalt, A.G. (1977) Deoxyribonucleic Acid Relatedness among Erwiniae and Other Enterobacteria II. Corn Stalk Rot Bacterium and Pectobacterium chrysanthemi. International Journal of Systematic and Evolutionary Microbiology, 27, 211-221.
http://dx.doi.org/10.1099/00207713-27-3-211 [9] Gallois, A., et al. (1992) Erwinia carotovora subsp. odorifera subsp. nov., Associated with Odorous Soft Rot of Chicory (Cichorium intybus L.). International Journal of Systematic and Evolutionary Microbiology, 42, 582-588.
http://dx.doi.org/10.1099/00207713-42-4-582 [10] Nassar, A., et al. (1994) Ribotyping of Erwinia chrysanthemi Strains in Relation to Their Pathogenic and Geographic Distribution. Applied and Environmental Microbiology, 60, 3781-3789. [11] Xiu, J.H., Ji, G.H., Wang, M., Yang, Y.L. and Li, C.Y. (2006) Molecular Identification and Genetic Diversity in Konnyaku’s Soft Rot Bacteria. Acta Microbiologica Sinica, 46, 522-525. [12] Hauben, L., Moore, E.R., Vauterin, L., Steenackers, M., Mergaert, J., Verdonck, L. and Swings, J. (1998) Phylogenetic Position of Phytopathogens within the Enterobacteriaceae. Systematic and Applied Microbiology, 21, 384-397.
http://dx.doi.org/10.1016/S0723-2020(98)80048-9 [13] Kwon, S.W., Go, S.J., Kang, H.W., Ryu, J.C. and Jo, J.K. (1997) Phylogenetic Analysis of Erwinia Species Based on 16S rRNA Gene Sequences. International Journal of Systematic Bacteriology, 47, 1061-1067.
http://dx.doi.org/10.1099/00207713-47-4-1061 [14] Dickey, R.S. (1979) Erwinia chrysanthemi: A Comparative Study of Phenotypic Properties of Strains from Several Hosts and Other Erwinia Species. Phytopathology, 69, 324-329.
http://dx.doi.org/10.1094/Phyto-69-324 [15] Dye, D.W. (1981) A Numerical Taxonomic Study of the Genus Erwinia. New Zealand Journal of Agricultural Research, 24, 223-229.
http://dx.doi.org/10.1080/00288233.1981.10420894 [16] Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes, M., et al. (1995) AFLP: A New Technique for DNA Fingerprinting. Nucleic Acids Research, 23, 4407-4414.
http://dx.doi.org/10.1093/nar/23.21.4407 [17] Geornaras, I., Kunene, N.F., von Holy, A. and Hastings, J.W. (1999) Amplified Fragment Length Polymorphism Fingerprinting of Pseudomonas Strains from a Poultry Processing Plant. Applied and Environmental Microbiology, 65, 3828-3833. [18] Janssen, P., Coopman, R., Huys, G., Swings, J., Bleeker, M., Vos, P., et al. (1996) Evaluation of the DNA Fingerprinting Method AFLP as an New Tool in Bacterial Taxonomy. Microbiology, 142, 1881-1893.
http://dx.doi.org/10.1099/13500872-142-7-1881 [19] Weisburg, W.G., Barns, S.M., Pelletier, D.A. and Lane, D.J. (1991) 16S Ribosomal DNA Amplification for Phylogenetic Study. Journal of Bacteriology, 173, 697-703. [20] Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor. [21] Tamura, K., Stecher, G., Peterson, D., Filipski, A. and Kumar, S. (2013) MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30, 2725-2729.
http://dx.doi.org/10.1093/molbev/mst197 [22] Han, Y.C., Teng, C.Z., Hu, Z.L. and Song, Y.C. (2008) An Optimal Method of DNA Silver Staining in Polyacrylamide Gels. Electrophoresis, 29, 1355-1358.
http://dx.doi.org/10.1002/elps.200700558 [23] Rohlf, F. (2000) NTSYS-pc Version 2.10 m, Numerical Taxonomy and Multivariate Analysis System (Computer Program). Exeter Software, Steauket. [24] Snijder, R.C., Cho, H.-R., Hendriks, M.M.W.B., Lindhout, P. and van Tuyl, J.M. (2004) Genetic Variation in Zantedeschia spp. (Araceae) for Resistance to Soft Rot Caused by Erwinia carotovora subsp. carotovora. Euphytica, 135, 119-128.
http://dx.doi.org/10.1023/B:EUPH.0000009546.88984.60