Volume 3, Issue 4, July 2015, Page: 30-36
Identification of Differentially Expressed Genes During Pseudomonas fluorescens Mediated Systemic Resistance in Cabbage
Kaunain Roohie, Department of Studies in Biotechnology, University of Mysore, Manasagangotri, Mysore, Karnataka, India
Sharanaiah Umesha, Department of Studies in Biotechnology, University of Mysore, Manasagangotri, Mysore, Karnataka, India
Received: Sep. 10, 2014;       Accepted: Oct. 31, 2014;       Published: Jul. 14, 2015
DOI: 10.11648/j.plant.20150304.11      View  3754      Downloads  67
Abstract
The study of microbial ecology and the microbial interactions with plants provides an insight into the biocontrol of plant diseases using antagonistic microbes. Pseudomonas fluorescens was used as a biological control agent against black rot disease caused by Xanthomonas campestris pv. campestris. The Suppression subtractive hybridization (SSH) was used to elucidate the differentially expressed genes in cabbage (Brassica oleracea var. capitata) upon the application of Pseudomonas fluorescens. A total of 140 expressed sequence tags (EST) were obtained. The analyses of these ESTs showed that many defense related genes like peroxidase, heat shock proteins, were upregulated. Many transcripts related to signalling pathways and pathogen recognition were identified. The important finding of the study is the identification of the unigene belonging to the SWEET protein family in cabbage. The study also resulted in the identification of 10 unigenes which possibly depict the interaction of Pseudomonas fluorescens in combating disease. These unigenes have been submitted to dbEST. The results show that those genes which are upregulated during pathogen attack are also induced upon application of Pseudomonas fluorescens indicating the possible mechanism of systemic resistance induced by P. fluorescens to combat disease.
Keywords
Xanthomonas campestris pv. campestris, RT-PCR, Pseudomonas Fluorescens, Suppression Subtractive Hybridization
To cite this article
Kaunain Roohie, Sharanaiah Umesha, Identification of Differentially Expressed Genes During Pseudomonas fluorescens Mediated Systemic Resistance in Cabbage, Plant. Vol. 3, No. 4, 2015, pp. 30-36. doi: 10.11648/j.plant.20150304.11
Reference
[1]
Alstrom S. (1991) Induction of disease resistance in common bean susceptible to halo blight pathogen after seed bacterization with rhizosphere pseudomonads. J Gen Appl Microbiol 37: 495-501.
[2]
Beattie GA, Lindow SE. (1995) The secret life of foliar bacterial pathogens on leaves. Ann Rev of Phytopathol 33:145–172.
[3]
Burr TJ, Matteson MC, Smith CA, Corral-Gracia MR,Huang TC. (1996) Effectiveness of bacteria and yeasts from apple orchards as biological control agents of apple scab. Biol Control 6:151–157.
[4]
Chen LQ, Hou BH, Lalonde S, Takanaga H, Hartung ML, Qu XQ, Guo WJ, Kim JG, Underwood W, Chaudhuri B, Chermak D, Antony G, White FF, Somerville SC, Mudgett MB, Frommer WB. (2010) Sugar transporters for intercellular exchange and nutrition of pathogens. Nature 468:527-532.
[5]
Das KK, Panda D, Nagaraju M, Sharma SG, Sarkar RK. (2004) Antioxidant enzymes and aldehyde releasing capacity of rice cultivars (Oryzae sativa L.) and determinants of anaerobic seedling establishment capacity. Bulg J Plant Physiol 30:34-44.
[6]
Diatchenko L, Lukyanov S, Lau YF, Siebert PD. (1999) Suppression subtractive hybridisation: a versatile method for identifying differentially expressed genes. Methods in Enzymol 303:349–380.
[7]
Ganeshmoorthi P, Anand T, Prakasan V, Bharani M, Ragupathi N, Samiyappan R. (2008) Plant growth promoting rhizobacterial (PGPR) bioconsortia mediates induction of defense-related proteins against infection of root rot pathogen in mulberry plants. J Plant Interact 3:233-244.
[8]
He CY, Hsiang T, Wolyn DJ. (2001) Activation of defense response to Fusarium infection in Asparagus densiflorus. Eur J Plant Pathol 107:473-483.
[9]
Hoffland E, Hakulinen J, Van Pelt JA. (1996) Comparison of systemic resistance induced by avirulent and nonpathogenic Pseudomonas species. Phytopathol 86:757-762.
[10]
International Seed Testing Authority. (2014) Detection of Xanthomonas campestris pv. campestris on Brassica spp. disinfested/disinfected seed with grinding.
[11]
Hutchison ML, Tester MA, and Gross DC. (1995) Role of biosurfactant and ion channel18 forming activities of syringomycin in transmembrane ion flux: a model for the mechanism of action in the plant pathogen interaction. Molec Plant-Microb Interac 8:610-620.
[12]
King EO, Ward MK, RaneyDE. (1956) Two simple media for the demonstration of pyocyanin and fluorescein. J Laboratory Clin Med 4:301-307.
[13]
Kurkcuoglu S, Degenhardt J, Lensing J, Abdul Nasser Al-Masri, Gau AE. (2007) Identification of differentially expressed genes in Malus domestica after application of the non-pathogenic bacterium Pseudomonas fluorescens Bk3 to the phyllosphere. J Experimental Botany 58:733–741
[14]
Lindow SE, Brandl MT. (2003) Microbiology of the phyllosphere. Appl Environ Microbiol 69:1875-1883.
[15]
Madamanchi NR and Alscher RG. (1991) Metabolic bases for differences in sensitivity of two pea cultivars to sulfur dioxide. Plant Physiol 97:88-93.
[16]
Massomo SMS, Mortensen CN, Mabagala RB, Newman MA, Hockenhull J. (2004) Biological control of Black Rot (Xanthomonas campestris pv campestris) of Cabbage in Tanzania with Bacillus strains. J Phytopathol 152:98–105.
[17]
Meena, B, Radhajeyalakshmi R, Marimuthu T, Vidhyasekaran P, Doraisamy S, Velazhahan R.(2000) Induction of pathogenesis related proteins, phenolics and phenylalanine ammonia lyase in groundnut by Pseudomonas fluorescens. J. Plant Dis. Protect 107:514 – 527.
[18]
Nandakumar R, Babu S, Viswanathan R, Raguchander T, Samiyappan R. (2001) Induction of systemic resistance in rice against sheath blight disease by Pseudomonas fluorescens. Soil Biol Biochem 33:603-612.
[19]
Ramamoorthy V, Raguchander T, Samiyappan R. (2002) Induction of defense-related proteins in tomato roots treated with Pseudomonas fluorescens Pf1 and Fusarium oxysporum f. sp. lycopersici. Plant Soil 239:55-68.
[20]
Schmidt C, Lorenz D, Wolf G (2001) Biological control of the grapevine dieback fungus Eutypa lata. Screening of bacterial antagonists. J Phytopathol 149:427–435.
[21]
Shruti Mishra, Naveen K Arora. (2012). Evaluation of rhizospheric Pseudomonas and Bacillus as biocontrol tool for Xanthomonas campestris pv campestris. World J Microbiol Biotechnol, 28:693–702.
[22]
Singh P, Piotrowski M, Kloppstech K, Gau AE (2004). Investigations on epiphytic living Pseudomonas species from Malus domestica with an antagonistic effect to Venturia inaequalis on isolated plant cuticle membranes. Environ Microbiol 11:1149–1158.
[23]
Staub T, Williams PH (1972) Factors influencing black rot lesion development in resistant and susceptible cabbage. Phytopathol 62:722–728.
[24]
Vanitha SC, Niranjana SR, Mortensen CN, Umesha S (2009) Bacterial wilt of tomato in Karnataka and its management by Pseudomonas fluorescens. Biocontrol 54:685-695
[25]
Wei G, Kloepper JW, Tuzun S. (1991) Induction of systemic resistance in cucumber to Colletotrichum orbiculare by selected strains of plant growth promoting rhizobacteria. Phytopathol 81:1508-1517.
[26]
Williams PH, Staub T, Sutton JC. (1972) Inheritance of black rot resistance in cabbage. Phytopathol 62:247–252
[27]
Williams PH. 1980. Black rot: a continuing threat to world crucifers. Plant Dis 64:736–742.
[28]
Zipfel C, Robatzek S, Navarro L, Oakeley EJ, Jones JDG, Felix G, Boller T. (2004). Bacterial disease resistance in Arabidopsis through flagellin perception. Nature 428:764–767.
Browse journals by subject