Abstract
The study aimed to reveal the diversity of endophytic bacteria in the roots of Chinese cabbage (CC) cultivated in two areas in Korea, namely, Seosang-gun (SS) and Haenam-gun (HN), and also in a transgenic plant (TP) from the laboratory. A total of 653 colonies were isolated from the interior of CC roots, comprising 118, 302, and 233 isolates from SS, HN, and TP samples, respectively. Based on 16S rRNA gene sequence analysis, the isolates belonged to four major phylogenetic groups: high-G+C Gram-positive bacteria (HGC-GPB), low-G+C Gram-positive bacteria (LGC-GPB), Proteobacteria, and Bacteriodetes. The most dominant groups in the roots of the SS, HN, and TP cultivars were LGC-GPB (48.3%), Proteobacteria (50.2%), and HGC-GPB (38.2%), respectively. Importantly, most of the isolates that produced cell-walldegrading enzymes belonged to the genus Bacillus. Bacillus sp. (HNR03, TPR06), Bacillus pumilus (SSR07, HNR11, TPR07), and Bacillus subtilis (TPR03) showed high antagonism against the tested food-borne pathogenic bacteria. In addition, Bacillus sp. (HNR03, TPR06), Bacillus pumilus (SSR07, HNR11, HNR17, TPR11), Microbacterium oxidans (SSR09, TPR04), Bacillus cereus HNR10, Pseudomonas sp. HNR13, and Bacillus subtilis (TPR02, TPR03) showed strong antagonistic activity against the fungi Phythium ultimum, Phytophthora capsici, Fusarium oxysporum, and Rhizoctonia solani. The endophytes isolated from the TP cultivar showed the strongest antagonistic reactions against pathogens. This study is the first report on endophytic bacteria from Chinese cabbage roots.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
An, D.J., Lew, K., and Lee, K.P. 1999. Effects of adipic acid and storage temperature on extending the shelf life of kimchi. Food Sci. Biotechnol. 8, 78–82.
Al-Mallah, M.K., Davey, M.R., and Cooking, E.C. 1987. Enzymatic treatment of clover root hairs removes a barrier to Rhizobium-host specificity. Nat. Biotechnol. 5, 1319–1322.
Andrade, L.A., de Souza, G.L.O.D., Nietsche, S., Xavier, A.A., Costa, M.R., Cardosa, A.S.M., Pereira, M.C.T., and Pereira, D.F.G.S. 2014. Analysis of the abilities of endophytic bacteria associated with banana tree roots to promote plant growth. J. Microbiol. 52, 27–34.
Barbosa, T.M., Serra, C.R., Ragione, R.M.L., Woodward, M.J., and Henriques, A.O. 2005. Screening for Bacillus isolates in the broiler gastrointestinal tract. Appl. Environ. Microbiol. 71, 968–978.
Berg, G., Krechel, A., Ditz, M., Sikora, R.A., Ulrich, A., and Hallmann, J. 2005. Endophytic and ectophytic potato-associated bacterial communities differ in structure and antagonistic function against plant pathogenic fungi. FEMS Microbiol. Ecol. 51, 215–229.
Cherif, M., Benhamou, N., and Belanger, R.R. 1993. Occurrence of cellulose and chitin in the hyphal walls of Pythium ultimum: a comparative study with other plant pathogenic fungi. Can. J. Microbiol. 39, 213–222.
Cho, K.M., Hong, S.Y., Lee, S.M., Kim, Y.H., Kahng, G.G., Lim, Y.P., Kim, H., and Yun, H.D. 2007. Endophytic bacterial communities in ginseng and their antifungal activity against pathogens. Microbial. Ecol. 54, 341–351.
Dalton, D.A., Kramer, S., Azios, N., Fusaro, S., Cahill, E., and Kennedy, C. 2004. Endophytic nitrogen fixation in dune grasses (Ammophila arenaria and Elymus mollis) from Oregon. FEMS Microbiol. Ecol. 49, 469–479.
de Melo Pereira, G.V., Magalhaes, K.T., Lorenzetii, E.R., Souza, T.P., and Schwan, R.F. 2012. A multiphasic approach for the identification of endophytic bacterial in strawberry fruit and their potential for plant growth promotion. Microb. Ecol. 63, 405–417.
Falcao, L.L., Silva-Werneck, J.O., Vilarinho, B.R., da Silva, J.P., Pomella, A.W.V., and Marcellino, L.H. 2014. Antimicrobial and plant growth-promoting properties of the cacao endophyte Bacillus subtilis ALB629. J. Appl. Microbiol. 116, 1584–1592.
Fukui, Y., Abe, M., Kobayashi, M., Yano, Y., and Satomi, M. 2014. Isolation of Hyphomonas strains that induce normal morphogenesis in protoplasts of the marine red algae Pyropia yezoensis. Microb. Ecol. 68, 556–566.
Haque, M.A., Lee, J.H., and Cho, K.M. 2015. Endophytic bacterial diversity in Korean kimchi made of Chinese cabbage leaves and their antimicrobial activity against pathogens. Food Cont. 56, 24–33.
Hong, Y., Yang, H.S., Chang, H.C., and Kim, H.Y. 2013. Comparison of bacterial community changes in fermenting kimchi at two different temperatures using denaturing gradient gel electrophoresis analysis. J. Microbiol. Biotechnol. 23, 76–84.
Islam, S.M.A., Math, R.K., Kim, J.M., Yun, M.G., Cho, J.J., Kim, E.J., Lee, Y.H., and Yun, H.D. 2010. Effect of plant age on endophytic bacterial diversity of balloon flower (Platycodon grandiflorum) root and their antimicrobial activities. Curr. Microbiol. 61, 346–356.
Jung, J.Y., Lee, S.H., and Jeon, C.O. 2014. Kimchi microflora: history, current status, and perspectives for industrial kimchi production. Appl. Microbiol. Biotechnol. 98, 2385–2393.
Jung, J.Y., Lee, S.H., Lee, H.J., Seo, H.Y., Park, W.S., and Jeon, C.O. 2012. Effects of Leuconostoc mesenteroides starter cultures on microbial communities and metabolities during kimchi fermentation. Int. J. Food Microbiol. 153, 378–387.
Khan, A.L., Waqas, M., Kang, S.M., Al-Harrasi, A., Hussain, J., Al-Rawahi, A., Al-Khiziri, S., Ullah, I., Ali, L., Jung, H.Y., et al. 2014. Bacterial endophyte Sphingomonas sp. LK11 produces gibberellins and IAA and promotes tomato plant growth. J. Microbiol. 52, 689–695.
Khastini, R.O., Ohta, H., and Narisawa, K. 2012. The role of a dark septate endophytic fungus, Veronaeopsis simplex Y34, in Fusarium disease suppression in Chinese cabbage. J. Microbiol. 50, 618–624.
Kim, M. and Chun, J. 2005. Bacterial community structure in kimchi, a Korean fermented vegetable food, as revealed by 16S rRNA gene analysis. Int. J. Food Microbiol. 103, 91–96.
Lacava, P.T., Araujo, W.L., Marcon, J., Maccheroni, W. Jr., and Azevedo, J.L. 2004. Interaction between endophytic bacteria from citrus plants and the phytopathogenic bacteria Xylella fastidiosa, causal agent of citrus-variegated chlorosis. Lett. Appl. Microbiol. 39, 55–59.
Lkeda, A.C., Bassani, L.L., Adamoski, D., Stringari, D., Cordeiro, V.K., Glienke, C., Steffens, M.B.R., Hungria, M., and Galli-Terasawa, L.V. 2013. Morphological and genetic characterization of endophytic bacteria isolated from roots of different maize genotypes. Microb. Ecol. 65, 154–160.
Li, C.H., Shi, L., Han, Q., Hu, H.L., Zhao, M.W., Tang, C.M., and Li, S.P. 2012. Biocontrol of verticillium wilt and colonization of cotton plants by an endophytic bacterial isolate. J. Appl. Microbiol. 113, 641–651.
Li, C.H., Zhao, M.W., Tang, C.M., and Li, S.P. 2010. Population dynamics and identification of endophytic bacteria antagonistic toward plant-pathogenic fungi in cotton root. Microb. Ecol. 59, 344–356.
Luo, Y., Sun, L., Zhu, Z., Ran, W., and Shen, Q. 2013. Identification and characterization of an antifungi Fusarium oxysporum f. sp. cucumerium protease from the Bacillus subtilis strain N7. J. Microbiol. 51, 359–366.
Marimuthu, S., Ramamoorthy, V., Samiyappan, R., and Subbian, P. 2013. Intercropping system with combined application of Azospirillum and Pseudomonas fluorescens reduces root rot incidence caused by Rhizoctonia bataticola and increases seed cotton yield. J. Phytopathol. 161, 405–411.
Moretti, M., Gilardi, G., Gullino, M.L., and Garibaldi, A. 2008. Biological control potential of Achromobacter xylosoxydans for suppressing Fusarium wilt of tomato. Int. J. Bot. 4, 369–375.
Ohike, T., Makuni, K., Okanami, M., and Ano, T. 2013. Screening of endophytic bacteria against fungal plant pathogens. J. Environ. Sci. 25, 122–126.
Ordentlich, A., Elad, Y., and Chet, I. 1988. The role of chitinase of Serratia marcescens in biocontrol of Sclerotium rolfsii. Phytopathology 78, 84–88.
Park, W.P. 1995. Effect of starch sources on yulmoo kimchi fermentation. Food Biotechnol. 4, 98–100.
Pathak, K.V. and Keharia, H. 2013. Characterization of fungal antagonistic bacilli isolated from aerial roots of banyan (Ficus benghalensis) using intact-cell MALDI-TOF mass spectrometry (ICMS). J. Appl. Microbiol. 114, 1300–1310.
Pereira, P., Nesci, A., and Etcheverry, M. 2007. Effects of biocontrol agents on Fusarium verticillioides count and fumonisin content in the maize agroecosystem: impact on rhizospheric bacterial and fungal groups. Biol. Control. 42, 281–287.
Prieto, P., Schiliro, E., Maldonado-Gonzalez, M.M., Valderrama, R., Barroso-Albarracin, J.B., and Mercado-Blanco, J. 2011. Root hairs play a key role in the endophytic colonization of olive roots by Pseudomonas spp. with biocontrol activity. Microb. Ecol. 62, 435–445.
Pu, Y., Sun, L., Wang, Y., Qi, D., Chen, D., Liu, H., Xu, D., Deng, C., and Li, J. 2013. Modeling inhibitory activity of a novel antimicrobial peptide AMPNT-6 from Bacillus subtilis against Vibrio parahaemolyticus in shrimp under various environmental conditions. Food. Control 33, 249–253.
Reinhold-Hurek, B. and Hurek, T. 2011. Living inside plants: bacterial endophytes. Curr. Opin. Plant. Biol. 14, 435–443.
Sakiyama, C.C.H., Paula, E.M., Pereira, P.C., Borges, A.C., and Silva, D.O. 2001. Characterization of pectin lyase produced by an endophytic strain isolated from coffee Cherries. Lett. Appl. Microbiol. 33, 117–121.
Sambrook, J. and Russel, D.W. 2001. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, N.Y., USA.
Santoyo, G., Orozco-Mosqueda, M.D.C., and Govindappa, M. 2012. Mechanisms of biocontrol and plant growth-promoting activity in soil bacterial species of Bacillus and Pseudomonas: a review. Biocontrol. Sci. Technol. 22, 855–872.
Schisler, D.A., Slininger, P.J., Behle, R.W., and Jackson, M.A. 2004. Formulation of Bacillus spp. for biological control of plant diseases. Phytopathology 94, 1267–1271.
Seghers, D., Wittebolle, L., Top, E.M., Verstraete, W., and Siciliano, S.D. 2004. Impact of agricultural practice on the Zea mays L. endophytic community. Appl. Environ. Microbiol. 70, 1475–1482.
Seo, W.T., Lim, W.J., Kim, E.J., Yun, H.D., Lee, Y.H., and Cho, K.M. 2010. Endophytic bacterial diversity in the young raddish and their antimicrobial activity against pathogens. J. Korean Soc. Appl. Biol. Chem. 53, 493–503.
Sessitsch, A., Reiter, B., and Berg, G. 2004. Endophytic bacterial communities of field-grown potato plants and their plant-growth promoting and antagonistic abilities. Can. J. Microbiol. 50, 239–249.
Strobel, G. and Daisy, B. 2003. Bioprospecting for microbial endophytes and their natural products. Microbiol. Mol. Biol. Rev. 67, 491–502.
Sturz, A.V. and Nowak, J. 2000. Endophytic communities of rhizobacter and the strategies required to create yield enhancing associations with crops. Appl. Soil Ecol. 15, 183–190.
Takahashi, M., Kita, Y., Kusaka, K., Mizuno, A., and Goto-Yamamoto, N. 2015. Evaluation of microbial diversity in the pilot-scale beer brewing process by culturedependent and culture-independent method. J. Appl. Microbiol. 118, 454–469.
Verma, S.C., Ladha, J.K., and Tripathi, A.K. 2001. Evaluation of plant growth promoting and colonization ability of endophytic diazotrophs from deep water rice. J. Biotechnol. 81, 127–141.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplemental material for this article may be found at http://www.springerlink.com/content/120956
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Haque, M.A., Yun, H.D. & Cho, K.M. Diversity of indigenous endophytic bacteria associated with the roots of Chinese cabbage (Brassica campestris L.) cultivars and their antagonism towards pathogens. J Microbiol. 54, 353–363 (2016). https://doi.org/10.1007/s12275-016-5641-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-016-5641-7