[en] Background
Some non-pathogenic rhizobacteria called Plant Growth Promoting Rhizobacteria (PGPR) possess the capacity to induce in plant defense mechanisms effective against pathogens. Precedent studies showed the ability of Pseudomonas putida BTP1 to induce PGPR-mediated resistance, termed ISR (Induced Systemic Resistance), in different plant species. Despite extensive works, molecular defense mechanisms involved in ISR are less well understood that in the case of pathogen induced systemic acquired resistance.
Results
We analyzed the activities of phenylalanine ammonia-lyase (PAL) and lipoxygenase (LOX), key enzymes of the phenylpropanoid and oxylipin pathways respectively, in tomato treated or not with P. putida BTP1. The bacterial treatment did not stimulate PAL activity and linoleate-consuming LOX activities. Linolenate-consuming LOX activity, on the contrary, was significantly stimulated in P. putida BTP1-inoculated plants before and two days after infection by B. cinerea. This stimulation is due to the increase of transcription level of two isoforms of LOX: TomLoxD and TomLoxF, a newly identified LOX gene. We showed that recombinant TomLOXF preferentially consumes linolenic acid and produces 13-derivative of fatty acids. After challenging with B. cinerea, the increase of transcription of these two LOX genes and higher linolenic acid-consuming LOX activity were associated with a more rapid accumulation of free 13-hydroperoxy-octadecatrienoic and 13-hydroxy-octadecatrienoic acids, two antifungal oxylipins, in bacterized plants.
Conclusion
In addition to the discovery of a new LOX gene in tomato, this work is the first to show differential induction of LOX isozymes and a more rapid accumulation of 13-hydroperoxy-octadecatrienoic and 13-hydroxy-octadecatrienoic acids in rhizobacteria mediated-induced systemic resistance.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Mariutto, Martin ; Université de Liège - ULiège > Département des sciences de la vie > Biologie moléculaire et biotechnologie végétales
Duby, Franceline ; Université de Liège - ULiège > Département des sciences de la vie > Génétique
Bent E. Induced systemic resistance mediated by plant growth-promoting Rhizobacteria (PGPR) and fungi (PGPF). Multigenic and induced systemic resistance in plants 2006, 225-258. New York, Springer US, Tuzun S, Bent E.
Van Peer R, Nieman GJ, Schippers B. Induced resistance and phytoalexin accumulation in biological control of Fusarium wilt of carnation by Pseudomonas sp. strain WCS417r. Phytopathology 1991, 81:728-734.
Pieterse CMJ, Van Wees SCM, Hoffland E, Van Pelt JA, Van Loon LC. Systemic resistance in Arabidopsis induced by biocontrol bacteria is independent of salicylic acid accumulation and pathogenesis-related gene expression. Plant Cell 1996, 8:1225-1237. 10.1105/tpc.8.8.1225, 161233, 8776893.
Van Loon LC, Bakker PAHM, Pieterse CMJ. Systemic resistance induced by rhizosphere bacteria. Annu Rev Phytopathol 1998, 36:453-483. 10.1146/annurev.phyto.36.1.453, 15012509.
Pieterse CMJ, Van Wees SCM, van Pelt JA, Knoester M, Laan R, Gerrits H, Weisbeek PJ, Van Loon LC. A novel signaling pathway controlling induced systemic resistance in Arabidopsis. The Plant Cell 1998, 10:1571-1580. 10.2307/3870620, 144073, 9724702.
Silva HSA, da Silva Romeiro R, Macagnan D, de Almeida Halfeld-Vieira B, Peirera MCB, Mounter A. Rhizobacterial induction of systemic resistance in tomato plants: non-specific protection and increase in enzyme activities. Biol Control 2004, 29:288-295.
Sailaja PR, Podile AR, Reddanna P. Biocontrol strain of Bacillus subtilis AF 1 rapidly induces lipoxygenase in groundnut (Arachis hypogaea L.) compared to crown rot pathogen Aspergillus niger. Eur J Plant Pathol 1997, 104:125-132.
Ferrie BJ, Beaudoin N, Burkhart W, Bowsher CG, Rothstein SJ. The cloning of two tomato lipoxygenase genes and their differential expression during fruit ripening. Plant Physiol 1994, 106:109-118. 10.1104/pp.106.1.109, 159505, 7972514.
Chen G, Hackett R, Walker D, Taylor A, Lin Z, Grierson D. Identification of a specific isoform of tomato lipoxygenase (TomloxC) involved in the generation of fatty acid-derived flavor compounds. Plant Physiol 2004, 136:1-11.
Krumbein A, Auerswald H. Characterization of aroma volatiles in tomatoes by sensory analyses. Nahrung 1998, 42:395-399. 10.1002/(SICI)1521-3803(199812)42:06<395::AID-FOOD395>3.3.CO;2-Z, 9881368.
Heitz T, Bergey DR, Ryan CA. A gene encoding a chloroplast-targeted lipoxygenase in tomato leaves is transiently induced by wounding, systemin, and methyl jasmonate. Plant Physiol 1997, 114:1085-1093. 10.1104/pp.114.3.1085, 158398, 9232884.
Prost I, Dhondt S, Rothe G, Vicente J, Rodriguez MJ, Kift N, Carbone F, Griffiths G, Esquerré-Tugayé MT, Rosahl S, Castresana C, Hamberg M, Fournier J. Evaluation of the antimicrobial activities of plant oxylipins supports their involvement in defense against pathogens. Plant Physiol 2005, 139:1902-1913. 10.1104/pp.105.066274, 1310568, 16299186.
Benhamou N, Kloepper JW, Quadt-Hallmann A, Tuzun S. Induction of defense-related ultrastructural modifications in pea root tissues inoculated with endophytic bacteria. Plant Physiol 1996, 112:919-929. 158019, 12226427.
Yedidia I, Shoresh M, Kerem Z, Benhamou N, Kapulnik Y, Chet I. Concomitant induction of systemic resistance to Pseudomonas syringae pv. lachrymans in cucumber by Trichoderma asperellum (T-203) and accumulation of phytoalexins. Appl Environ Microb 2003, 69:7343-7353.
Jacques P, Ongena M, Gwose I, Seinsche D, Schroder H, Delfosse P, Thonart P, Taraz K, Budzikiewicz H. Structure and characterization of isopyoverdin from Pseudomonas putida BTP1 and its relation to the biogenetic pathway leading to pyoverdins. Z Naturforsch 1995, 50:622-629.
Ongena M, Daayf F, Jacques P, Thonart P, Benhamou N, Paulitz TC, Bélanger RR. Systemic induction of phytoalexins in cucumber in response to treatments with fluorescent pseudomonads. Plant Pathol 2000, 49:523-530.
Ongena M, Duby F, Rossignol F, Fauconnier ML, Dommes J, Thonart P. Stimulation of the lipoxygenase pathway is associated with systemic resistance induced in bean by a nonpathogenic Pseudomonas Strain. Mol Plant Microbe 2004, 17:1009-1018.
Adam A, Ongena M, Duby F, Dommes J, Thonart P. Systemic resistance and lipoxygenase-related defense response induced in tomato by Pseudomonas putida strain BTP1. BMC Plant Biol 2008, 8:113. 10.1186/1471-2229-8-113, 2596797, 19000301.
Royo J, Vancanneyt G, Pérez A G, Sanz C, Störmann K, Rosahl S, Sánchez-Serrano JJ. Characterization of three potato lipoxygenases with distinct enzymatic activities and different organ-specific and wound-regulated expression patterns. J Biol Chem 1996, 271:21012-21019. 10.1074/jbc.271.35.21012, 8702864.
Halitschke R, Baldwin IT. Antisense LOX expression increases herbivore performance by decreasing defense responses and inhibiting growth-related transcriptional reorganization in Nicotiana attenuata. Plant J 2003, 36:794-807. 10.1046/j.1365-313X.2003.01921.x, 14675445.
Prigge ST, Boyington JC, Faig M, Doctor KS, Gaffney BJ, Amzel LM. Structure and mechanism of lipoxygenases. Biochimie 1997, 79:629-636. 10.1016/S0300-9084(97)83495-5, 9479444.
Feussner I, Wasternack C. The lipoxygenase pathway. Annu Rev Plant Biol 2002, 53:275-297. 10.1146/annurev.arplant.53.100301.135248, 12221977.
Liavonchanka A, Feussner I. Lipoxygenases: Occurrence, functions and catalysis. J Plant Physiol 2006, 163:348-357. 10.1016/j.jplph.2005.11.006, 16386332.
Bell E, Mullet JE. Characterization of an Arabidopsis Lipoxygenase Gene Responsive to Methyl Jasmonate and Wounding. Plant Physiol 1993, 103:1133-1137. 10.1104/pp.103.4.1133, 159098, 8290626.
Zheng SJ, van Dijk JP, Bruinsma M, Dicke M. Sensitivity and speed of induced defense of cabbage (Brassica oleracea L.): dynamics of BoLOX expression patterns during insect and pathogen attack. Mol Plant Microbe 2007, 20:1332-1345.
Leon J, Royo J, Vancanneyt G, Sanz C, Silkowski H, Griffiths G, Sanchez-Serrano JJ. Lipoxygenase H1 gene silencing reveals a specific role in supplying fatty acid hydroperoxides for aliphatic aldehyde production. J Biol Chem 2002, 277:416-423. 10.1074/jbc.M107763200, 11675388.
Keegstra K, Olsen LJ, Theg SM. Chloroplastic precursors and their transport across the envelope membranes. Annu Rev Plant Phys 1989, 40:471-501.
Bell E, Creelman RA, Mullet JE. A chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in Arabidopsis. P Natl Acad Sci USA 1995, 92:8675-8679.
Farmaki T, Sanmarti{dotless}n M, Jimenez P, Paneque M, Sanz C, Vancanneyt G, Leon J, Sanchez-Serrano JJ. Differential distribution of the lipoxygenase pathway enzymes within potato chloroplasts. J Exp Bot 2007, 58:555-568. 10.1093/jxb/erl230, 17210991.
Porta H, Figueroa-Balderas R E, Rocha-Sosa M. Wounding and pathogen infection induce a chloroplast-targeted lipoxygenase in the common bean (Phaseolus vulgaris L.). Planta 2008, 227:363-373. 10.1007/s00425-007-0623-y, 17899174.
Ramamoorthy V, Raguchander T, Samiyappan R. Enhancing resistance of tomato and hot pepper to Pythium diseases by seed treatment with fluorescent Pseudomonads. Eur J Plant Pathol 2002, 108:429-441.
Blée E. Phytooxylipins and plant defense reactions. Prog Lipid Res 1998, 37:33-72.
Wasternack C, Stenzel I, Hause B, Hause G, Kutter C, Maucher H, Neumerkel J, Feussner I, Miersch O. The wound response in tomato - Role of jasmonic acid. J Plant Physiol 2006, 163:297-306. 10.1016/j.jplph.2005.10.014, 16368162.
Halitschke R, Ziegler J, Keinänen M, Baldwin IT. Silencing of hydroperoxide lyase and allene oxide synthase reveals substrate and defense signaling crosstalk in Nicotiana attenuate. The Plant J 2004, 40:35-46.
Royo J, Leon J, Vancanneyt G, Albar JP, Rosahl S, Ortego F, Castanera P, Sanchez-Serrano JJ. Antisense-mediated depletion of a potato lipoxygenase reduces wound induction of proteinase inhibitors and increases weight gain of insect pests. P Natl Acad Sci USA 1999, 96:1146-1151.
Matsui K, Miyahara C, Wilkinson J, Hiatt B, Knauf V, Kajiwar T. Fatty Acid Hydroperoxide Lyase in Tomato Fruits: Cloning and Properties of a Recombinant Enzyme Expressed in Escherichia coli. Biosc Biotechnol Biochem 2000, 64:1189-1196.
Bate NJ, Rothstein SJ. C6-volatiles derived from the lipoxygenase pathway induce a subset of defense-related genes. The Plant J 1998, 16:561-569.
Schaller F. Enzymes of the biosynthesis of octadecanoid-derived signalling molecules. J Exp Bot 2001, 52:11-23. 10.1093/jexbot/52.354.11, 11181709.
Adam A, Jourdan E, Ongena M, Duby F, Dommes J, Thonart P. Resistance induced in cucumber and tomato by a non-pathogenic Pseudomonas putida strain. Parasitica 2005, 61:13-22.
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K. Phenol/SDS method for plant RNA preparation. Current protocols in molecular biology 1987, 4.3.1-4.3.4. Wiley and Sons. New York.
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG. Clustal W and Clustal X version 2.0. Bioinformatics 2007, 23:2947-2948. 10.1093/bioinformatics/btm404, 17846036.
http://www.ebi.ac.uk/Tools/clustalw2/index.html
Chevenet F, Brun C, Banuls A L, Jacq B, Chisten R. TreeDyn: towards dynamic graphics and annotations for analyses of trees. BMC Bioinformatics 2006, 7:439. 10.1186/1471-2105-7-439, 1615880, 17032440.
Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, Chevenet F, Dufayard JF, Guindon S, Lefort V, Lescot M, Claverie JM, Gascuel O. Phylogeny.fr: robust phylogenetic nalysis for the non-specialist. Nucleic Acids Res 2008, 36.
http://www.cbs.dtu.dk/services/ChloroP/
Emanuelsson O, Nielsen H, Brunak S, von Heijne G. Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 2000, 300:1005-1016. 10.1006/jmbi.2000.3903, 10891285.
http://wolfpsort.org/
http://hc.ims.u-tokyo.ac.jp/iPSORT/
Bannai H, Tamada Y, Maruyama O, Nakai K, Miyano S. Extensive feature detection of N-terminal protein sorting signals. Bioinformatics 2002, 18:298-305. 10.1093/bioinformatics/18.2.298, 11847077.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970, 227:680-685. 10.1038/227680a0, 5432063.
Boutaud O, Brash AR. Purification and catalytic activities of the two domains of the allene oxide synthase-lipoxygenase fusion protein of the coral Pleuxaura homomalla. The J of Biol Chem 1999, 274:33764-33770.
Fauconnier ML, Rojas-Beltran J, Dupuis B, Delaplace P, Frettinger P, Gosset V, du Jardin P. Changes in oxylipin synthesis after Phytophtora infestans infection of potato leaves do not correlate with resistance. Plant Physiol Bioch 2008, 46:823-831.
Fauconnier ML, Delcarte J, Hoyaux P, du Jardin P, Marlier M. Potato tubers exhibit both hemolytic and heterolytic hydroperoxide fatty acid-cleaving activities. Bioch Soc 2000, 28:853-855.
