Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Tomato mitogen activated protein kinases regulate the expression of extracellular invertase Lin6 in response to stress related stimuli

Tae Kyung Hyun A , Anja Hoffmann A , Alok K. Sinha B C and Thomas Roitsch A
+ Author Affiliations
- Author Affiliations

A Julius-von-Sachs-Institut fuer Biowissenschaften, Universitaet Wuerzburg, Julius-von-Sachs-Platz 2, 97082 Wuerzburg, Germany.

B National Institute of Plant Genome Research, PB No. 10531, Aruna Asaf Ali Road, New Delhi 110067, India.

C Corresponding author. Email: alok_sinha@nipgr.res.in

Functional Plant Biology 36(12) 1088-1097 https://doi.org/10.1071/FP09136
Submitted: 4 June 2009  Accepted: 31 August 2009   Published: 3 December 2009

Abstract

Activation of mitogen-activated protein kinases (MAPKs) is a common reaction of plant cells in defence-related signal transduction pathways. Since the downstream events after the activation of MAPKs are largely unknown in plants, the role of MAPKs in the coordinate regulation of defence reactions and primary carbon metabolism by stress related stimuli has been analysed in tomato (Lycopersicon peruvianum Mill.). Thus, the relationship between MAPK, LpMPK2 and LpMPK3 and extracellular invertase Lin6, as the key enzyme of an apoplasmic phloem unloading pathway, has been analysed. It was observed that the mRNAs of LpMPK3 and Lin6 are sequentially induced by the same set of stress related stimuli, wounding, a fungal elicitor derived from Fusarium oxysporum lycopersici, the endogenous plant derived elicitor PGA and salt stress, while LpMPK2 transcripts are constitutively expressed. In a gain of function approach, a His-tagged version of LpMPK2 and a HA-tagged version of LpMPK3 were transiently and functionally expressed in leaves of transgenic tobacco (Nicotiana tabacum L.) plants expressing the β-glucuronidase reporter gene under control of the Lin6 promoter via agro-infection. The induction of the Lin6 promoter, as revealed by an increase in β-glucuronidase activity after 24 h, was dependent both on the expression and activation of both LpMPK2 and LpMPK3. These data suggest that the induction of extracellular invertase Lin6 by stress-related stimuli requires LpMPK2 and LpMPK3, and thus demonstrate that MAPK signalling might be involved in the regulation of primary carbon metabolism in general and sink metabolism in particular.

Additional keywords: GUS activity, Lycopersicon peruvianum, MAPK, sink metabolism.


Acknowledgements

TKH acknowledges the fellowship provided by DAAD and Graduierten Kolleg 1342. AKS and TR thanks funding from joint Indo-German DAAD-DST project (No. D/07/13475 and DST/INT/DAAD/P-165/2007).


References


Andreasson E, Jenkins T, Brodersen P, Thorgrimsen S, Petersen NHT, Zhu S, Qiu J, Micheelsen P, Rocher A, Petersen M (2005) The MAP kinase substrate MKS1 is a regulator of plant defense responses. The EMBO Journal 24, 2579–2589.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Asai T, Tena G, Plotnikova J, Willmann MR, Chiu WL, Gomez-Gomez L, Boller T, Ausubel FM, Sheen J (2002) MAP kinase signaling cascade in Arabidopsis innate immunity. Nature 415, 977–983.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Beimen A, Witte L, Barz W (1992) Growth characteristics and elicitor-induced reactions of photosynthetically active and heterotrophic cell suspension cultures of Lycopersicon peruvianum Mill. Botanica Acta 105, 152–160. open url image1

Berger S, Sinha AK, Roitsch T (2007) Plant physiology meets phytopathology: plant primary metabolism and plant–pathogen interactions. Journal of Experimental Botany 58, 4019–4026.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Bogre L, Ligterink W, Meskiene L, Barker PJ, Heberle-Bors E, Huskisson NS, Hirt H (1997) Wounding induces the rapid and transient activation of a specific MAPK pathway. The Plant Cell 9, 75–83.
Crossref | PubMed |
open url image1

Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Chang LF, Karin M (2001) Mammalian MAP kinase signaling cascades. Nature 410, 37–40.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Cho JI, Lee SK, Ko S, Kim HK, Jun SH , et al. (2005) Molecular cloning and expression analysis of the cell-wall invertase gene family in rice (Oryza sativa L.). Plant Cell Reports 24, 225–236.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Chomczynski P, Sacchi N (1987) Single step method of RNA isolation by acid guanidium thyocynate-phenol extraction. Analytical Biochemistry 162, 156–159.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Dahan J, Pichereau C, Rossignol M, Blanc S, Wendehenne D, Pugin A, Bourque S (2009) Activation of a nuclear-localized SIPK in tobacco cells challenged by cryptogein, an elicitor of plant defence reactions. The Biochemical Journal 418, 191–200.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Davis R (2000) Signal transduction by the JNK group of MAP kinases. Cell 103, 239–252.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Ehness R, Ecker M, Godt D, Roitsch T (1997) Glucose and stress independently regulate source/sink relations and defense mechanisms via signal transduction pathways involving protein phosphorylation. The Plant Cell 9, 1825–1841.
Crossref | PubMed |
open url image1

Godt DE, Roitsch T (1997) Regulation and tissue-specific distribution of mRNAs for three extracellular invertase isoenzymes of tomato suggests an important function in establishing and maintaining sink metabolism. Plant Physiology 115, 273–282.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Goetz M, Godt DE, Guivarc’h A, Kahmann U, Chriqui D, Roitsch T (2001) Induction of male sterility in plants by metabolic engineering of the carbohydrate supply. Proceedings of the National Academy of Sciences of the United States of America 98, 6522–6527.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Hamel LP, Nicole M, Sritubtim S, Morency M, Ellis M, Ehlting J, Beaudoin N, Barbazuk B, Klessig D, Lee J (2006) Ancient signals: comparative genomics of plant MAPK and MAPKK gene families. Trends in Plant Science 11, 192–198.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Hirose T, Takano M, Terao T (2002) Cell wall invertase in developing rice caryopsis: molecular cloning of OsCIN1 and analysis of its expression in relation to its role in grain filling. Plant & Cell Physiology 43, 452–459.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Holley SR, Yalamanchili RD, Moura DS, Ryan CA, Stratmann JW (2003) Convergence of signaling pathways induced by systemin, oligosaccharide elicitors, and ultraviolet-B radiation at the level of mitogen-activated protein kinases in Lycopersicon peruvianum suspension-cultured cells. Plant Physiology 132, 1728–1738.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Ichimura K, Mizoguchi T, Yoshida R, Yuasa T, Shinozaki K (2000) Various abiotic stresses rapidly activate Arabidopsis MAP kinases ATMPK4 and ATMPK6. The Plant Journal 24, 655–665.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Jonak C, Kiegerl S, Ligterink W, Barker PJ, Huskisson NS, Hirt H (1996) Stress signaling in plants: a mitogen-activated protein kinase pathway in activated by cold and drought. Proceedings of the National Academy of Sciences of the United States of America 93, 11 274–11 279.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Joo S, Liu Y, Lueth A, Zhang S (2008) MAPK phosphorylation-induced stabilization of ACS6 protein is mediated by the non-catalytic C-terminal domain, which also contains the cis-determinant for rapid degradation by the 26S proteasome pathway. The Plant Journal 54, 129–140.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Kandoth PK, Ranf S, Pancholi SS, Jayanty S, Walla MD, Miller W, Howe GA, Lincoln DE, Stratmann JW (2007) Tomato MAPKs LeMPK1, LeMPK2, and LeMPK3 function in the systemin-mediated defense response against herbivorous insects. Proceedings of the National Academy of Sciences of the United States of America 104, 12 205–12 210.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Koch KE (1996) Carbohydrate-modulated gene expression in plants. Annual Review of Plant Physiology and Plant Molecular Biology 47, 509–540.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Kyriakis JM, Avruch J (1996) Protein kinase cascades activated by stress and inflammatory cytokines. BioEssays 18, 567–577.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Link V, Hofmann MG, Sinha AK, Ehness R, Strnad M, Roitsch T (2002a) Biochemical evidence for the activation of distinct subsets of mitogen-activated protein kinases by voltage and defense-related stimuli. Plant Physiology 128, 271–281.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Link V, Sinha AK, Vasishta P, Hofmann MG, Proels RK, Ehness R, Roitsch T (2002b) A heat activated MAP kinase in tomato: a possible regulator of heat response. FEBS Letters 531, 179–183.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Liu Y, Zhang S (2004) Phosphorylation of 1-aminocyclopropane-1-carboxylic acid synthase by MPK6, a stress responsive mitogen-activated protein kinase, induces ethylene biosynthesis in Arabidopsis. The Plant Cell 16, 3386–3399.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

MAPK Group (2002) Mitogen-activated protein kinase cascades in plants: a new nomenclature. Trends in Plant Science 7, 301–308.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Mayrose M, Bonshtien A, Sessa G (2004) LeMPK3 is a mitogen-activated protein kinase with dual specificity induced during tomato defense and wounding responses. The Journal of Biological Chemistry 279, 14 819–14 827.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Menke FLH, Kang H, Chen Z, Park JM, Kumar D, Klessig DF (2005) Tobacco transcription factor WRKY1 is phosphorylated by the MAP kinase SIPK and mediates HR-like cell death in tobacco. Molecular Plant-Microbe Interactions 18, 1027–1034.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Miao Y, Laun TM, Smykowski A, Zentgraf U (2007) Arabidopsis MEKK1 can take a short cut: it can directly interact with senescence-related WRKY53 transcription factor on the protein level and can bind to its promoter. Plant Molecular Biology 65, 63–76.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Mishra NS, Tuteja R, Tuteja H (2006) Signaling through MAP kinase networks in plant. Archives of Biochemistry and Biophysics 452, 55–68.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Pedley KF, Martin GB (2004) Identification of MAPKs and their possible MAPK kinase activators involved in the pto-mediated defense response of tomato. The Journal of Biological Chemistry 279, 49 229–49 235.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Proels RK, Roitsch T (2009) Extracellular invertase LIN6 of tomato: a pivotal enzyme for integration of metabolic, hormonal, and stress signals is regulated by a diurnal rhythm. Journal of Experimental Botany 60, 1555–1567.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Proels R, Hause B, Berger S, Roitsch T (2003) Tissue specific expression in sexual organs and novel modes of hormonal regulation of tomato invertase tandem genes. Plant Molecular Biology 52, 191–201.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Proels RK, González MC, Roitsch T (2006) Gibberellins dependent induction of tomato extracellular invertase Lin7 is required for pollen development. Functional Plant Biology 33, 547–554.
Crossref | GoogleScholarGoogle Scholar | open url image1

Roitsch T (1999) Source–sink regulation by sugar and stress. Current Opinion in Plant Biology 2, 198–206.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Roitsch T, Gonzalez MC (2004) Function and regulation of plant invertases: sweet sensations. Trends in Plant Science 9, 606–613.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Roitsch T, Bittner M, Godt DC (1995) Induction of apoplastic invertase of Chenopodium rubrum by D-glucose and a glucose analogue and tissue specific expression suggest a role in sink source regulation. Plant Physiology 108, 285–294.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Roitsch T, Balibrea ME, Hofmann M, Proels R, Shina AK (2003) Extracellular invertase: key metabolic enzyme and PR protein. Journal of Experimental Botany 54, 513–524.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Romeis T, Piedras P, Zhang S, Klessig DF, Hirt H, Jones JDG (1999) Rapid Avr9- and Cf-9-dependent activation of MAP kinases in tobacco cell cultures and leaves: convergence of resistance gene, elicitor, wound, and salicylate responses. The Plant Cell 11, 273–287.
Crossref | PubMed |
open url image1

Sinha AK, Hofmann MG, Römer U, Köckenberger W, Elling L, Roitsch T (2002) Metabolizable and non-metabolizable sugars activate different signal transduction pathways in tomato. Plant Physiology 128, 1480–1489.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Stulemeijer IJE, Stratmann JW, Joosten MHAJ (2007) Tomato mitogen-activated protein kinases LeMPK1, LeMPK2, and LeMPK3 are activated during the Cf-4/Avr4-induced hypersensitive response and have distinct phosphorylation specificities. Plant Physiology 144, 1481–1494.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Sturm A, Chrispeels MJ (1990) cDNA cloning of carrot extracellular β-fructosidase and its expression in response to wounding and bacterial infection. The Plant Cell 2, 1107–1119.
Crossref | PubMed |
open url image1

Tena G, Asai T, Chiu WL, Sheen J (2001) Plant mitogen activated protein kinase signaling cascades. Current Opinion in Plant Biology 4, 392–400.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Tymowska-Lalanne Z, Kreis M (1998) Expression of the Arabidopsis thaliana invertase gene family. Planta 207, 259–265.
Crossref | GoogleScholarGoogle Scholar | open url image1

Widmann C, Gibson S, Jarpe MB, Johnson GL (1999) Mitogen-activated protein kinase: conservation of a three-kinase module from yeast to human. Physiological Reviews 79, 143–180. open url image1

Yoo SD, Cho YH, Tena G, Xiong Y, Sheen J (2008) Dual control of nuclear EIN3 by bifurcate MAPK cascades in C2H4 signaling. Nature 451, 789–795.
Crossref | GoogleScholarGoogle Scholar | open url image1

Zeng Y, Wu Y, Avigne WT, Koch KE (1999) Rapid repression of maize invertases by low oxygen. Invertase/sucrose synthase balance, sugar signaling potential, and seedling survival. Plant Physiology 121, 599–608.
Crossref | GoogleScholarGoogle Scholar | open url image1

Zhang S, Klessig DF (1997) Salicylic acid activates a 48-KD MAP kinase in tobacco. The Plant Cell 9, 809–824.
Crossref | PubMed |
open url image1

Zhang S, Klessig DF (1998a) Resistance gene N-mediated de novo synthesis and activation of a tobacco mitogen-activated protein kinase by tobacco mosaic virus infection. Proceedings of the National Academy of Sciences of the United States of America 95, 7433–7438.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Zhang S, Klessig DF (1998b) The tobacco wounding-activated mitogen-activated protein kinase is encoded by SIPK. Proceedings of the National Academy of Sciences of the United States of America 95, 7225–7230.
Crossref | GoogleScholarGoogle Scholar | open url image1

Zhang S, Liu Y (2001) Activation of salicylic acid-induced protein kinase, a mitogen-activated protein kinase, induces multiple defense responses in tobacco. The Plant Cell 13, 1877–1889.
Crossref | PubMed |
open url image1

Zhang S, Du H, Klessig DF (1998c) Activation of the tobacco SIP kinase by both a cell wall-derived carbohydrate elicitor and purified proteinaceous elicitins form Phytophthora spp. The Plant Cell 10, 435–450.
Crossref |
open url image1