Vegetos
(An International Journal of Plant Research & Biotechnology)
Isolation and characterization of Xanthomonas campestris pv. campestris from cabbage and its Inhibition using endophytic bacteria
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Research Articles | Published: 23 June, 2025
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Keywords: Black rot, n Xanthomonas campestris pv. campestrisn , Pathogenicity, Endophytic bacteria, Inhibitory effect
Abstract
The plant pathogenic bacterium Xanthomonas campestris pv. campestris (Xcc) is a seed borne bacterium that causes black rot of cabbage and causes huge loss in yield. In the present study Xcc were isolated from cabbage and characterized by morphological features, biochemical tests, pathogenicity test and molecular analysis. A total of five Xcc isolates were obtained from five different cabbage fields around Mysuru region of Karnataka. When cultured on YDC media, the isolates showed typical yellow, raised, translucent, mucoid colonies. The biochemical analysis showed that the bacteria were negative for Gram’s staining, oxidase reaction and nitrate reduction, and positive for starch hydrolysis, gelatin hydrolysis, KOH solubility, catalase test, indole production and H2S production. In pathogenicity test, all the isolates showed typical black rot symptoms characterized by V-shaped lesions that progressed from yellow to brown along the leaf margins, accompanied by the blackening of veins. The molecular identification of the isolates was carried out by amplification of 16s rRNA region and hrpF gene. Further, endophytic bacteria were isolated from cabbage and screened for their ability to inhibit Xcc. A total of five endophytic bacteria were obtained and among them four isolates showed inhibitory effect on Xcc. The isolate P3 showed maximum inhibition with inhibition zone of 28 ± 0. 14 mm and was identified as Bacillus safensis by amplification of 16s rRNA region. The results provide insights on characterization of Xcc and indicate that Bacillus safensis could play an important role in biocontrol of Xcc on further characterization.
References
Ab Rahman SFS, Singh E, Pieterse CM, Schenk PM (2018) Emerging microbial biocontrol strategies for plant pathogens. Plant Sci 267:102–111
Afzal M, Khan QM, Sessitsch A (2014) Endophytic bacteria: prospects and applications for the phytoremediation of organic pollutants. Chemosphere 117:232–242
Alvarez AM (2000) Black rot of crucifers. Mechanisms of resistance to plant diseases. Springer Netherlands, Dordrecht, pp 21–52
Barzman M, Bàrberi P, Birch ANE, Boonekamp P, Dachbrodt-Saaydeh S, Graf B, Sattin M (2015) Eight principles of integrated pest management. Agron Sustain Dev 35:1199–1215
Bhore SJ, Ravichantar N, Loh CY (2010) Screening of endophytic bacteria isolated from leaves of SambungNyawa [Gynuraprocumbens (Lour.) Merr.] forcytokinin-like compounds. Bioinformation 5(5):191
Boer SD, Elphinstone JG, Saddler GS (2007) Molecular detection strategies for phytopathogenic bacteria. Biotechnology and plant disease management. CAB International, Wallingford UK, pp 165–194
Burlakoti RR, Chen JR, Hsu CF, Burlakoti P, Kenyon L (2018) Molecular characterization, comparison of screening methods, and evaluation of cross-pathogenicity of black rot (Xanthomonas Campestris pv. Campestris) strains from cabbage, Choy sum, leafy mustard and Pakchoi from Taiwan. Plant Pathol 67(7):1589–1600
Büttner D, Noël L, Stuttmann J, Bonas U (2007) Characterization of the non-conserved hpaB-hrpF region in the hrp pathogenicity Island from Xanthomonas campestris pv. vesicatoria. Mol Plant Microbe Interact 20(9):1063–1074
Chidamba L, Bezuidenhout CC (2012) Characterization of Xanthomonas campestris pv. campestris isolates from South Africa using genomic DNA fingerprinting and pathogenicity tests. Eur J Plant Pathol 133:811–818
Compant S, Duffy B, Nowak J, Clément C, Barka EA (2005) Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Appl Environ Microbiol 71(9):4951–4959
de Almeida Lopes KB, Carpentieri-Pipolo V, Fira D, Balatti PA, López SMY, Oro TH, Degrassi G (2018) Screening of bacterial endophytes as potential biocontrol agents against soybean diseases. J Appl Microbiol 125(5):1466–1481
Eljounaidi K, Lee SK, Bae H (2016) Bacterial endophytes as potential biocontrol agents of vascular wilt diseases–review and future prospects. Biol Control 103:62–68
Eshel D, Regev R, Orenstein J, Droby S, Gan-Mor S (2009) Combining physical, chemical and biological methods for synergistic control of postharvest diseases: A case study of black root rot of Carrot. Postharvest Biol Technol 54(1):48–52
Fadiji AE, Babalola OO (2020) Elucidating mechanisms of endophytes used in plant protection and other bioactivities with multifunctional prospects. Front Bioeng Biotechnol 8:467
Fathi Z, Rezaei R, Charehgani H, Ghaderi F, Ghalavoz MG (2024) Biocontrol potential of epiphytic bacteria against Xanthomonas citri pathotypes A and A. Egypt J Biol Pest Control 34(1):33
Feng Y, Zhang Y, Shah OU, Luo K, Chen Y (2023) Isolation and identification of endophytic bacteria Bacillus sp. ME9 that exhibits biocontrol activity against Xanthomonas phaseoli pv. manihotis. Biology 12(9):1231
Goss M, Mafongo P, Gubba A, Sam T (2017) Black rot (Xanthomonas Campestris pv. Campestris) control in field grown cabbage (Brassica Oleracea var. Sugar Loaf) with Moringa oleifera extracts. Int J Plant Soil Sci 18(2):1–11
Grenni P, Ancona V, Caracciolo AB (2018) Ecological effects of antibiotics on natural ecosystems: A review. Microchem J 136:25–39
Gupta M, Vikram A, Bharat N (2013) Black rot-A devastating disease of crucifers: a review. Agricultural Reviews 34(4):269–278
Hong CE, Jo SH, Moon JY, Lee JS, Kwon SY, Park JM (2015) Isolation of novel leaf-inhabiting endophytic bacteria in Arabidopsis thaliana and their antagonistic effects on phytophathogens. Plant Biotechnol Rep 9:451–458
Hugouvieux V, Barber CE, Daniels MJ (1998) Entry of Xanthomonas campestris pv. campestris into hydathodes of Arabidopsis thaliana leaves: a system for studying early infection events in bacterial pathogenesis. Mol Plant Microbe Interact 11(6):537–543
Icoz SM, Polat I, Sulu G, Yilmaz M, Unlu A, Soylu S, Baysal Ö (2014) First report of bacterial blight of pomegranate caused by Xanthomonas axonopodis pv. punicae in Turkey. Plant Dis 98(10):1427–1427
Islam MN, Ali MS, Choi SJ, Hyun JW, Baek KH (2019) Biocontrol of citrus canker disease caused by Xanthomonas citri subsp. citri using an endophytic Bacillus thuringiensis. Plant Pathol J 35(5):486
Janda JM, Abbott SL (2007) 16S rRNA gene sequencing for bacterial identification in the diagnostic laboratory: pluses, perils, and pitfalls. J Clin Microbiol 45(9):2761–2764
Jelušić A, Popović T, Dimkić I, Mitrović P, Peeters K, Višnjevec AM, Berić T (2021) Changes in the winter oilseed rape Microbiome affected by Xanthomonas campestris pv. campestris and biocontrol potential of the Indigenous Bacillus and Pseudomonas isolates. Biol Control 160:104695
Lacava PT, Araújo WLD, Marcon J, Maccheroni W Jr, Azevedo JLD (2004) Interaction between endophytic bacteria from citrus plants and the phytopathogenic bacteria xylella fastidiosa, causal agent of citrus-variegated chlorosis. Lett Appl Microbiol 39(1):55–59
Lema M, Cartea ME, Sotelo T, Velasco P, Soengas P (2012) Discrimination of Xanthomonas campestris pv. campestris races among strains from Northwestern Spain by Brassica spp. Genotypes and rep-PCR. Eur J Plant Pathol 133:159–169
Liu Z, Wang H, Wang J, Lv J, Xie B, Luo S, Yu J (2022) Physical, chemical, and biological control of black rot of brassicaceae vegetables: A review. Front Microbiol 13:1023826
Lodewyckx C, Vangronsveld J, Porteous F, Moore ER, Taghavi S, Mezgeay M, der Lelie DV (2002) Endophytic bacteria and their potential applications. CRC Crit Rev Plant Sci 21(6):583–606
Massomo SM, Mabagala RB, Mortensen CN, Hockenhull J, Swai IS (2005) Cabbage production in tanzania: challenges faced by smallholder farmers in the management of black rot disease. J Sustainable Agric 26(4):119–141
Massomo SM, Nielsen H, Mabagala RB, Mansfeld-Giese K, Hockenhull J, Mortensen CN (2003) Identification and characterization of Xanthomonas campestris pv. campestris strains from Tanzania by pathogenicity tests, biolog, rep-PCR and fatty acid Methyl ester analysis. Eur J Plant Pathol 109:775–789
Mateus JR, Dal’Rio I, Jurelevicius D, da Mota FF, Marques JM, Ramos RTJ, Seldin L (2021) Bacillus velezensis T149-19 and Bacillus safensis T052-76 as potential biocontrol agents against foot rot disease in sweet potato. Agriculture 11(11):1046
Mengistu AA (2020) Endophytes: colonization, behavior, and their role in defense mechanism. Int J Microbiol 2020(1):6927219
Moreb N, Murphy A, Jaiswal S, Jaiswal AK (2020) Cabbage. Nutritional composition and antioxidant properties of fruits and vegetables: 33–54
Park YJ, Lee BM, Ho-Hahn J, Lee GB, Park DS (2004) Sensitive and specific detection of Xanthomonas campestris pv. campestris by PCR using species-specific primers based on HrpF gene sequences. Microbiol Res 159(4):419–423
Popović T, Balaž J, Ignjatov M, Mitrović P, Gavrilović V, Jošić D (2014) Identification and genetic characterisation of Xanthomonas campestris pv. campestris as an oilseed rape pathogen in Serbia. J Plant Pathol:96(3)
Ray LR, Alam MS, Junaid M, Ferdousy S, Akter R, Hosen SZ, Mouri NJ (2021) Brassica oleracea var. capitata f. alba: A review on its botany, traditional uses, phytochemistry and Pharmacological activities. Mini Rev Med Chem 21(16):2399–2417
Rabbee MF, Ali MS, Baek KH (2019) Endophyte Bacillus velezensis isolated from Citrus spp. Controls streptomycin-resistant Xanthomonas citri subsp. citri that causes citrus bacterial canker. Agronomy 9(8):470
Rekik H, Jaouadi NZ, Gargouri F, Bejar W, Frikha F, Jmal N, Jaouadi B (2019) Production, purification and biochemical characterization of a novel detergent-stable Serine alkaline protease from Bacillus safensis strain RH12. Int J Biol Macromol 121:1227–1239
Selim HM, Gomaa NM, Essa AM (2016) Antagonistic effect of endophytic bacteria against some phytopathogens. Egypt J Bot 1:74–81
Singh D, Dhar S, Yadava DK (2011) Genetic and pathogenic variability of Indian strains of Xanthomonas campestris pv. campestris causing black rot disease in crucifers. Curr Microbiol 63:551–560
Singh D, Rathaur PS, Vicente JG (2016) Characterization, genetic diversity and distribution of Xanthomonas campestris pv. campestris races causing black rot disease in cruciferous crops of India. Plant Pathol 65(9):1411–1418
Singh Dinesh, Dhar SHRI, Yadav DK (2010) Effect of endophytic bacterial antagonists against black rot disease of cauliflower caused by Xanthomonas campestris pv. campestris. Indian Phytopathol 63(2):122–126
Sugio A, Yang B, White FF (2005) Characterization of the HrpF pathogenicity Peninsula of Xanthomonas oryzae pv. oryzae. Mol Plant Microbe Interact 18(6):546–554
Taheri E, Tarighi S, Taheri P (2023) An endophytic bacterium with biocontrol activity against important wheat pathogens. Biol Control 183:105243
Tang YW, Stratton CW, Aslanzadeh J (2006) Biochemical profile-based microbial identification systems. Adv Techniques Diagn Microbiol: 84–116
Ullah A, Nisar M, Ali H, Hazrat A, Hayat K, Keerio AA, Yang X (2019) Drought tolerance improvement in plants: an endophytic bacterial approach. Appl Microbiol Biotechnol 103:7385–7397
Uuh-Narvaez JJ, Segura‐Campos MR (2021) Cabbage (Brassica Oleracea var. capitata): A food with functional properties aimed to type 2 diabetes prevention and management. J Food Sci 86(11):4775–4798
Van den Mooter M, Swings J (1990) Numerical analysis of 295 phenotypic features of 266 Xanthomonas strains and related strains and an improved taxonomy of the genus. Int J Syst Evol MicroBiol 40(4):348–369
Vendan RT, Yu YJ, Lee SH, Rhee YH (2010) Diversity of endophytic bacteria in ginseng and their potential for plant growth promotion. J Microbiol 48:559–565
Vicente JG, Conway J, Roberts SJ, Taylor JD (2001) Identification and origin of Xanthomonas campestris pv. campestris races and related pathovars. Phytopathology 91(5):492–499
Vicente JG, Holub EB (2013) Xanthomonas campestris pv. campestris (cause of black rot of crucifers) in the genomic era is still a worldwide threat to brassica crops. Mol Plant Pathol 14(1):2–18
Wang X, Liang L, Shao H, Ye X, Yang X, Chen X, Wang J (2022) Isolation of the novel strain Bacillus amyloliquefaciens F9 and identification of lipopeptide extract components responsible for activity against Xanthomonas citri subsp. Citri Plants 11(3):457
Wu PS, Liu CH, Hu SY (2021) Probiotic Bacillus safensis NPUST1 administration improves growth performance, gut microbiota, and innate immunity against Streptococcus iniae in nile tilapia (Oreo chromisniloticus). Microorganisms 9(12):2494
Wu T, Xu J, Liu J, Guo WH, Li XB, Xia JB, Wang RQ (2019) Characterization and initial application of endophytic Bacillus safensis strain ZY16 for improving phytoremediation of oil-contaminated saline soils. Front Microbiol 10:991
Wulff EG, Mguni CM, Mortensen CN, Keswani CL, Hockenhull J (2002) Biological control of black rot (Xanthomonas Campestris pv. Campestris) of brassicas with an antagonistic strain of Bacillus subtilis in Zimbabwe. Eur J Plant Pathol 108:317–325
Author Information
Department of Studies in Biotechnology, University of Mysore, Manasagangotri, Musuru, India