Abstract
The tomato (Lycopersicon esculentum Mill.) endo-β-1,4-glucanase (EGase) Cel1 protein was characterized in fruit using specific antibodies. Two polypeptides ranging between 51 and 52 kDa were detected in the pericarp, and polypeptides ranging between 49 and 51 kDa were detected in locules. The polypeptides recognized by Cel1 antiserum in fruit are within the size range predicted for Cel1 protein and could be derived from heterogeneous glycosylation. Cel1 protein accumulation was examined throughout fruit ripening. Cel1 protein appears in the pericarp at the stage in which many ripening-related changes start, and remains present throughout fruit ripening. In locules, Cel1 protein is already present at the onset of fruit ripening and remains constant during fruit ripening. This pattern of expression supports a possible role for this EGase in the softening of pericarp tissue and in the liquefaction of locules that takes place during ripening. The accumulation of Cel1 protein was also analyzed after fungal infection. Cel1 protein and mRNA levels are down-regulated in pericarp after Botrytis cinerea infection but are not affected in locular tissue. The same behavior was observed when fruits were infected with Penicillium expansum, another fungal pathogen. Cel1 protein and mRNA levels do not respond to wounding. These results support the idea that the tomato Cel1 EGase responds to pathogen infection and supports a relationship between EGases, plant defense responses and fruit ripening.
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Abbreviations
- EGase :
-
Endo-β-1,4-glucanase
- PG :
-
Polygalacturonase
References
Bennett AB, DellaPenna D (1987) Polygalacturonase gene expression in ripening tomato fruit. In: Nevins D, Jones R (eds) Tomato biotechnology. Liss, New York, pp 299–308
Bowles DJ (1990) Defence-related proteins in higher plants. Annu Rev Biochem 59:873–907
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254
Brady CJ, MacAlpine G, McGlasson W, Ueda Y (1982) Polygalacturonase in tomato fruits and the induction of ripening. Aust J Plant Physiol 9:171–178
Brady CJ, McGlasson W, Pearson J, Meldrum S, Kopeliovitch E (1985) Interaction between the amount and molecular forms of polygalacturonase, calcium and firmness in tomato fruit. J Am Soc Hort Sci 110:254–258
Brummell DA, Catalá C, Lashbrook C, Bennett AB (1997) A membrane-anchored E-type endo 1,4-β-glucanase is localized on Golgi and plasma membranes of higher plants. Proc Natl Acad Sci USA 94:4794–4799
Bugbee DJ (1993) A pectin lyase inhibitor protein from cell walls of sugar beet. Phytopathology 83:63–68
Catala C, Rose JKC, Bennett AB (1997) Auxin regulation and spatial localization of an endo-1,4-β-d-glucanase and a xyloglucan endotransglycosylase in expanding tomato hypocotyls. Plant J 12:417–426
Company P, González-Bosch C (2003) Identification of a copper chaperone from tomato fruits infected with Botrytis cinerea by differential display. Biochem Biophys Res Commun 304:825–830
Cooper W, Bouzayen M, Hamilton A, Barry C, Grierson D (1998) Use of transgenic plants to study the role of ethylene and polygalacturonase during infection of tomato fruit by Colletotrichum gloeosporoides. Plant Pathol 47:308–316
del Campillo E, Bennett AB (1996) Pedicel breakstrength and cellulase gene expression during tomato flower abscission. Plant Physiol 111:813–820
DellaPenna D, Alexander DC, Bennett AB (1986) Molecular cloning of tomato fruit polygalacturonase: analysis of polygalacturonase mRNA levels during ripening. Proc Natl Acad Sci USA 83:6420–6424
Fischer RL, Bennett AB (1991) Role of cell wall hydrolases in fruit ripening. Annu Rev Plant Physiol Plant Mol Biol 42:675–703
González-Bosch C, Brummel DA, Bennett AB (1996) Differential expression of endo-1,4-β-glucanase genes in pericarp and locules of wild type and mutant tomato fruit. Plant Physiol 111:1313–1319
González-Bosch C, del Campillo E, Bennett AB (1997) Immunodetection and characterization of tomato endo-β-1,4-glucanase Cel1 protein in flower abscission zones. Plant Physiol 114:1541–1546
Hall CB (1964) Cellulase activity in tomato fruits according to portion and maturity. Bot Gaz 125:156–157
Hobson GE (1968) Cellulase activity during the maturation and ripening of tomato fruit. J Food Sci 33:588–592
Huber DJ (1985) A possible role for cellulose and Cx-cellulase activity in locular gel formation in tomato fruit. HortScience 20:442–443
Huber DJ, Lee JH (1986) Comparative analysis of pectins from pericarp and locular gel in developing tomato fruit. ACS Symp Ser 310:141–156
Kitajima S, Sato F (1999) Plant pathogenesis-related proteins: molecular mechanisms of gene expression and protein function. J Biochem 125:1–8
Kramer M, Sanders R, Bolkan H, Waters C, Sheehy RE, Hiatt WR (1992) Post harvest evaluation of transgenic tomatoes with reduced levels of polygalacturonase: processing, firmness and disease resistance. Postharvest Biol Technol 1:241–255
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lashbrook CC, González-Bosch C, Bennett AB (1994) Two divergent endo-β-1,4-glucanase genes exhibit overlapping expression in ripening fruit and abscising flowers. Plant Cell 6:1485–1493
Lashbrook CC, Giovannoni JJ, Hall BD, Fischer RL, Bennett AB (1998) Transgenic analysis of tomato endo-β-1,4-glucanase gene function. Role of cel1 in floral abscission Plant J 13:303–310
Maclachlan B, Brady C (1992) Multiple forms of 1,4-β-glucanase in ripening tomato fruits include a xyloglucanase activatable by xyloglucan oligosaccharides. Aust J Plant Physiol 19:137–146
MacLauchlan WR, García-Conesa MT, Williamson G, Rza M, Ravenstein P, Maat J (1999) A novel class of protein from wheat which inhibits xylanases. Biochem J 338:441–446
Milligan B, Gasser CS (1995) Nature and regulation of pistil-expressed genes in tomato. Plant Mol Biol 28:691–711
Rose JKC, Bennett AB (1999) Cooperative disassembly of the cellulose–xyloglucan network of plant cell walls: parallels between cell expansion and fruit ripening. Trends Plant Sci 4:176–183
Sobotka FE, Watada AE (1971) Cellulase in high pigment and crimson tomato fruit. J Am Soc Hort Sci 96:705–707
Stotz HU, Powell ALT, Damon SE, Greve LC, Bennett AB, Labavitch JM (1993) Molecular characterization of a polygalacturonase inhibitor from Pyrus communis L. cv. Bartlett. Plant Physiol 102:133–138
Teixidó N, Viñas I, Usall J, Magan N (1998) Control of blue mold of apples by preharvest application of Candida sake grown in media with different water activity. Phytopathology 88:960–964
Vogel J, Raab T, Schiff C, Somerville S (2002) PMR6, a pectate lyase-like gene required for powdery mildew susceptibility in Arabidopsis. Plant Cell 14:2095–2106
Walton JD (1994) Deconstructing the cell wall. Plant Physiol 104:1113–1118
Acknowledgments
The authors are grateful to Dr. Juan P. Navarro-Aviñó for advice with preparation of figures. We also thank Hector Olivas for assistance with ethylene determinations. This work was supported by grants from Plan Nacional I+D (ALI97-0846-C02-01 and AGL2003-08481). P.C. was the recipient of a long-term predoctoral fellowship (grant ALI97-0846-C02).
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Real, M.D., Company, P., García-Agustín, P. et al. Characterization of tomato endo-β-1,4-glucanase Cel1 protein in fruit during ripening and after fungal infection. Planta 220, 80–86 (2004). https://doi.org/10.1007/s00425-004-1321-7
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DOI: https://doi.org/10.1007/s00425-004-1321-7