iTRAQ Proteins Analysis of Early Infected Papaya Plants with Papaya Dieback Pathogen

  • Norliza . A.B Department of Biochemistry UPM


Papaya dieback disease is characterized by the greasy water-soaked lesions and spots on leaves and crowns. Defoliation and blemished of the papaya fruits are also being observed as a result of infection by Erwinia mallotivora. In an attempt to understand the molecular mechanisms leading to the bacteria pathogenesis and the papaya plant response to infection in a compatiblereaction, protein profiling during 24 hours post infection was studied using iTRAQ mass spectrometry analysis. The bacterium was sprayed into wounded leaves of a susceptible papaya cultivar (Eksotika 1) and proteome analysis was performed. The comparison of protein patterns of the treated and the control plants were carried out by labelling the control sample with iTRAQ 8 plex reagent 113 and inoculated samples with the iTRAQ 8 plex reagent 115 whichwere then analysed by peptide mass fingerprinting and identified by searches in public databases. Biochemical changes occurring in infected tissues were observed. Among the differentially expressed proteins were enolase, maturase K, superoxide dismutase, ascorbate peroxidase, phosphoribulokinase, CT114 and a hypothetical protein with unknown function.


[1] Anon. FAOSTAT. Agricultural production. Crops primary
(papaya). 2006. Retrieved in 2006 from
[2] Noriha, M. A., Hamidun, B. Rohaiza, A. R. and Indu Bala S. J.
Erwinia mallotivora sp., a new phatogen of papaya (Carica papaya)
in Peninsular Malaysia. Int J Mol Sci. 2011;12:39-45.
[3] Webb, R. R. Epidemiology and control of bacterial canker of
papaya caused by an Erwinia sp. on St. Croix. U.S. Virgin Islands.
Plant Dis. 1985;69(4):305-309.
[4] Maktar, N.H., Kamis, S., Mohd Yusof, F.Z. and Hussain, N.H.
Erwinia papaya causing papaya dieback in Malaysia. New Disease
Reports. Plant Pathol. 2008;57(4):774.
[5] Von Rantt, A. Ubereine Bakteriendekranheit be idem
Melonenbaume (Carica papaya L.) auf Java. Zentbl Bakteriol
Parasitenkd Infektrankh Hyg. 1931;84:418-487 (in German, as
quoted by Gardan et al. 2004).
[6] Trujillo, E. E. and Schroth, M. N. Two bacterial diseases of papaya
trees cause by Erwinia species in the northern Mariana Islands.
Plant Dis. 1982;66:116-120.
[7] Guevara, Y., Rondon, A., Maselli, A., Salcedo, F. and Betancourt, J.
Marchitez bacteriana del lechosero Carica papaya L. en Venezuela.
Agronomia Trop. 1993;43:107-116 (in Spanish, as quoted by
Mohamed and Wan Kelthom, 2005).
[8] Gardan, L., Christen, R., Achouk, W. and Prior, P. Erwinia sp.
Nov., A pathogen of papaya (Carica papaya). Int J Syst Evol
Microbiol. 2004;54:107-113.
[9] Fukushima A, Kusano M, Redestig H, Arita M, Saito K. Integrated
omics approaches in plant systems biology. Curr Opin Chem Biol,
[10] Weckwerth W. Green systems biology: From single genomes,
proteomes and metabolomes to ecosystems research and
biotechnology. J Proteomics. 2011;75:284–305.
[11] Zieske L. R. A perspective on the use of iTRAQ reagent technology
for protein complex and profiling studies. J Exp Bot. 2006;57:1501–
[12] Ross PL, Huang YN, Marchese JN. Multiplexed protein
quantitation in Saccharomyces cerevisiae using amine-reactive
isobaric tagging reagents. Mol Cell Proteomics. 2004;3:1154–1169.
[13] Van Loon L.C., Rep M., Pieterse C.M.J. Significance of inducible
defense-related proteins in infected plants. Ann Rev Phytopath.
[14] Chan, Y.K. Backcross method in improvement of papaya (Carica
papaya L.). National Symposium on Genetic & Breeding of Crops
and Animals.11-13 November 1986, UKM, Bangi. Malays Appl
Biol. 1987;16: 95-100.
[15] Schneider, D.J. and Collmer, A. Studying Plant-Pathogen
Interactions in the Genomics Era: Beyond Molecular Koch's
Postulates to Systems Biology. Ann Rev Phytopathol. 2010;48(1):
[16] Wang, W.R. Scali, V. and Cresti, M. A universal and rapid protocol
for protein extraction from recalcitrant plant tissues for proteomic
analysis. Electrophoresis. 2006;27:2782–2786.
[17] Compton, S.J. and jones, C.G. Mechanism of dye response and
interference in the Bradford protein assay. Anal Biochem.
[18] Fukao, T., Kennedy, R. A., Yamasue, Y., & Rumpho, M. E. Genetic
and biochemical analysis of anaerobically induced enzymes during
seed germination of Echinochloa crus-galli varieties tolerant and
intolerant of anoxia. J Exp Bot. 2003;54:1421–1429.
[19] Gottig, N., Garavaglia, B.S, Daurelio, L.D, Valentine, A., Gehring,
C., Orellano, E.G. Modulating host homeostasis as a strategy in the
plant-pathogen arms race. Commun Integr Biol. 2009;2:89 – 90.
[20] Garavaglia, B.S., Thomas, L., Gottig, N., Dunge, G., Garofalo, C.G,
Daurelio L.D. A eukaryotic acquired gene by a biotrophic
phytopathogen allows prolonged survival on the host by
counteracting the shut-down of plant photosynthesis. PLoS ONE.
2010;5: e8950.
[21] Bonfig, K.B., Schreiber, U., Gabler, A., Roitsch, T., Berger, S.
Infection with virulent and avirulent P. syringae strains
differentially affects photosynthesis and sink metabolism in
Arabidopsis leaves. Planta 2006;225: 1–12.
[22] Chou H, Bundock N, Rolfe S, Scholes J. Infection of Arabidopsis
thaliana leaves with Albugo candida causes a reprogramming of
host metabolism. Mol Plant Pathol. 2000;1: 99-113.
[23] Swarbrick PJ, Schulze-Lefert P, Scholes JD. Metabolic
consequences of susceptibility and resistance (race-specific and
broad-spectrum) in barley leaves challenged with powdery mildew.
Plant Cell Environ. 2006;29:1061–1076.
[24] Perez-Bueno ML, Ciscato M, VandeVen M, Garcia-Luque I,
Valcke R, Baron M. Imaging viral infection: Studies on Nicotiana
benthamiana plants infected with the pepper mild mottle tobamo
virus. Photosynth Res. 2006;90:111-123.
[25] Cernadas RA, Camillo LR, Benedetti CE. Transcriptional analysis
of the sweet orange interaction with the citrus canker pathogens
Xanthomonas axonopodispv. citri and Xanthomonas axonopodis pv.
aurantifolii. Mol Plant Pathol. 2008;9:609–631.
[26] Campos, A., Gonçalo da Costa, G.D., Coelho, A.V., Fevereiro, P.
Identification of bacterial protein markers and enolase as a plant
response protein in the infection of Oleaeuropaea subsp. europaea
by Pseudomonas savastanoipv. Savastanoi. Eur J Plant Pathol.
2009;125: 603–616.
[27] Scott, K. J., Craigie, J. S., & Smillie, R. M. Pathways of respiration
in plant tumors. Plant Physiol. 1964;39:323–327.
[28] Kollipara, K. P., Saab, I. N., Wych, R. D., Lauer, M. J., &
Singletary, G. W. Expression profiling of reciprocal maize hybrids
divergent for cold germination and desiccation tolerance. Plant
Physiol. 2002;129: 974–992.
[29] Pandey SP, Somssich IE. The role of WRKY transcription factors in
plant immunity. Plant Physiol. 2009;150:1648–1655.
[30] Zhang A, Jiang M, Zhang J, Tan M, Hu X. Mitogen-activated
protein kinase is involved in abscisic acid-induced antioxidant
defense and acts downstream of reactive oxygen species production
in leaves of maize plants. Plant Physiol.2006;141:475–487.
[31] Neill SJ, Desikan R, Hancock JT. Hydrogen peroxide signaling.
Curr Opin Plant Biol. 2002;5:388–395.
[32] Mittler R. Oxidative stress, antioxidants and stress tolerance. Trends
in Plant Science. 2002;7:405–410.
[33] Park SY, Ryu SH, Jang IC, Kwon SY, Kim JG, Kwak SS.
Molecular cloning of a cytosolic ascorbate peroxidase cDNA from
cell cultures of sweetpotato and its expression in response to stress.
Molecular Genetics and Genomics. 2004;271:339–346.
[34] Solomon M, Belenghi B, Delledonne M, Menachem E, Levine A.
The involvement of cysteine proteases and protease inhibitor genes
in the regulation of programmed cell death in plants. Plant Cell.
[35] Beers, E.P., Jones, A.M., Dickerman, A.W. The S8 serine, C1A
cysteine and A1 aspartic protease families in Arabidopsis.
Phytochemistry. 2004;65:43–58.
[36] Cai M, Qiu D, Yuan T, Ding X, Li H, Duan L, Xu C, Li X, Wang S.
Identification of novel pathogen-responsive cis-elements and their
binding proteins in the promoter of OsWRKY13, a gene regulating
rice disease resistance. Plant Cell Environ. 2008;31:86–96.
[37] Pandey, S.P. and Somssich, I.E. The role of WRKY transcription
factors in plant immunity. Plant Physiol. 2009;150:1648–1655.
[38] Zhou W, Eudes F, Laroche A.Identification of differentially
regulated proteins in response to a compatible interaction between
the pathogen Fusarium graminearum and its host, Triticum
aestivum. Proteomics. 2006;6:4599–4609.
[39] Kesari R, Trivedi PK, Nath P. Gene expression of pathogenesisrelated
protein during banana ripening and after treatment with 1-
MCP. Postharvest Biol Technol. 2010;56:64-70.
[40] Borad V and S. Sriram. Pathogenesis-Related Proteins for the Plant
Protection. Asian J Exp Sci. 2008;22(3):189-196.
How to Cite
A.B, Norliza .. iTRAQ Proteins Analysis of Early Infected Papaya Plants with Papaya Dieback Pathogen. Asian Journal of Plant Biology, [S.l.], v. 3, n. 1, p. 1-7, mar. 2016. ISSN 2289-5868. Available at: <>. Date accessed: 21 sep. 2018.