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

  • Norliza . A.B Biotechnology Research Centre, MARDI, P. O. Box 12301, 50774 Kuala Lumpur, Malaysia
  • S. NormahfuzaHusna Horticulture Research Centre, G.P.O. Box 12301, 50774 Kuala Lumpur, Malaysia.
  • B. Rafidah Biotechnology Research Centre, MARDI, P. O. Box 12301, 50774 Kuala Lumpur, Malaysia
  • N.A. Shaharuddin Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, UPM 43400 Serdang, Selangor, Malaysia
  • L. Rozeita Horticulture Research Centre, G.P.O. Box 12301, 50774 Kuala Lumpur, Malaysia.
Keywords: Papaya, Erwinia mallotivora, protein profiling, iTRAQ mass spectrometry

Abstract

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.

References

Anon. FAOSTAT. Agricultural production. Crops primary

(papaya). 2006. Retrieved in 2006 from http://apps.fao.org/faostat/

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.

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.

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.

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).

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.

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).

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.

Fukushima A, Kusano M, Redestig H, Arita M, Saito K. Integrated

omics approaches in plant systems biology. Curr Opin Chem Biol,

;13:532–538.

Weckwerth W. Green systems biology: From single genomes,

proteomes and metabolomes to ecosystems research and

biotechnology. J Proteomics. 2011;75:284–305.

Zieske L. R. A perspective on the use of iTRAQ reagent technology

for protein complex and profiling studies. J Exp Bot. 2006;57:1501–

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.

Van Loon L.C., Rep M., Pieterse C.M.J. Significance of inducible

defense-related proteins in infected plants. Ann Rev Phytopath.

;44:135-162.

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.

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):

-479.

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.

Compton, S.J. and jones, C.G. Mechanism of dye response and

interference in the Bradford protein assay. Anal Biochem.

;151:369-374.

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.

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.

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.

;5: e8950.

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.

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.

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.

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.

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.

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.

;125: 603–616.

Scott, K. J., Craigie, J. S., & Smillie, R. M. Pathways of respiration

in plant tumors. Plant Physiol. 1964;39:323–327.

doi:10.1104/pp.39.3.323.

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.

Pandey SP, Somssich IE. The role of WRKY transcription factors in

plant immunity. Plant Physiol. 2009;150:1648–1655.

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.

Neill SJ, Desikan R, Hancock JT. Hydrogen peroxide signaling.

Curr Opin Plant Biol. 2002;5:388–395.

Mittler R. Oxidative stress, antioxidants and stress tolerance. Trends

in Plant Science. 2002;7:405–410.

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.

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.

;11:431–443.

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.

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.

Pandey, S.P. and Somssich, I.E. The role of WRKY transcription

factors in plant immunity. Plant Physiol. 2009;150:1648–1655.

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.

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.

Borad V and S. Sriram. Pathogenesis-Related Proteins for the Plant

Protection. Asian J Exp Sci. 2008;22(3):189-196.

Published
2015-07-30
Section
Articles