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A 54-kDa polypeptide identified by 2D-PAGE and bulked segregant analysis underlies differences for pH values in tomato fruit

Abstract

Fruit pH is an important quality attribute in tomato and it is defined during ripening. The aims of this work were to detect pericarp polypeptides associated with pH in an interspecific tomato BC1 generation by 1D-PAGE and to identify those differentially expressed polypeptides by comparing 2D-PAGE protein profiles from bulked segregant analysis (BSA). Polypeptide patterns were resolved by 1D-PAGE in a BC1 population obtained by crossing the cv. ‘Caimanta’ of Solanum lycopersicum (recurrent parental genotype) and the accession LA722 of S. pimpinellifolium (donor parental genotype). Putative QTL for fruit quality were detected by single point analysis. The presence of a 54-kDa band at the mature green stage (MG) carried by the wild genotype decreased the mean value of the pH trait. A BSA combined with 2D-PAGE was applied to the extreme phenotypes for pH in the BC1 segregating population. Four differentially expressed spots were detected when the polypeptide patterns of the bulks were compared. The spots had the expected molecular mass (around 54-kDa), and they were present in the lower-pH bulk and absent in the higher-pH one. The spots were identified by MS MALDI-TOF and two of them showed homology with the ATP synthase CF1 alpha subunit of S. lycopersicum. These results indicate that the association between the polypeptide marker and a fruit quantitative trait detected by 1D-PAGE not only would indicate genetic linkage but also could be directly related with the gene underlying the quantitative trait.

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Abbreviations

1D-PAGE:

One dimensional gel electrophoresis

2D-PAGE:

Two dimensional gel electrophoresis

BSA:

Bulked segregant analysis

kDa:

Kilo Dalton

MG:

Mature green stage

RR:

Red ripe stage

SDS–PAGE:

Sodium dodecyl sulphate–polyacrylamide gel electrophoresis

References

  1. Almeida DPF, Huber DJ (1999) Apoplastic pH and inorganic ion levels in tomato fruit: a potential means for regulation of cell wall metabolism during ripening. Physiol Plant 105:506–512

    CAS  Article  Google Scholar 

  2. Centeno DC, Osorio S, Nunes-Nesi A, Bertolo ALF, Carneiro RT, Araújo WL, Steinhauser M-C, Michalska J, Rohrmann J, Geigenberger P, Oliver SN, Stitt M, Carrari F, Rose JKC, Fernie AR (2011) Malate plays a crucial role in starch metabolism, ripening, and soluble solid content of tomato fruit and affects postharvest softening. Plant Cell 23:162–184

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. Chayut N, Yuan H, Ohali S, Meir A, Yeselson Y, Portnoy V, Zheng Y, Fei Z, Schaffer A (2015) A bulk segregant transcriptome analysis reveals metabolic and cellular processes associated with orange allelic variation and fruit β-carotene accumulation in melon fruit. BMC Plant Biol 15:1–18

    Article  Google Scholar 

  4. Etienne A, Génard M, Lobit P, Bugaud C (2013) What controls fleshy fruit acidity? A review of malate and citrate accumulation in fruit cells. J Exp Bot 64:1451–1469

    CAS  Article  PubMed  Google Scholar 

  5. Faurobert M, Mihr C, Bertin N, Pawlowski T, Negroni L, Sommerer N, Causse M (2007) Major proteome variations associated with cherry tomato pericarp development and ripening. Plant Phys 143:1327–1346

    CAS  Article  Google Scholar 

  6. Fulton TM, Bucheli P, Voirol E, Lopez J, Petiard V, Tanksley SD (2002) Quantitative trait loci (QTL) affecting sugars, organic acids and other biochemical properties possibly contributing to flavor, identified in four advanced backcross populations of tomato. Euphytica 127:163–177

    CAS  Article  Google Scholar 

  7. Gallo M, Rodríguez GR, Zorzoli R, Pratta GR (2011) Ligamiento genético entre variables asociadas a calidad del fruto de tomate y polipéptidos expresados en dos estados de madurez. Rev Fac Cs Agr 43:145–156

    Google Scholar 

  8. Giovannoni JJ (2004) Genetic regulation of fruit development and ripening. Plant Cell 16:170–180

    Article  Google Scholar 

  9. Görg A, Weiss W, Dunn MJ (2004) Current two-dimensional electrophoresis technology for proteomics. Proteomics 4:3665–3685

    Article  PubMed  Google Scholar 

  10. Grierson D, Tucker G (1983) Timing of ethylene and polygalacturonase synthesis in relation to the control of tomato fruit ripening. Planta 157:174–179

    CAS  Article  PubMed  Google Scholar 

  11. Jones RA, Scott SJ (1983) Improvement of tomato flavor by genetically increasing sugar and acid contents. Euphytica 32:845–855

    Article  Google Scholar 

  12. Liang XQ, Luo M, Holbrook CC, Guo BZ (2006) Storage protein profiles in Spanish and runner market type peanuts and potential markers. BMC Plant Biol 6:24

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. Liu BH (1998) Statistical genomics: linkage, mapping, and QTL analysis. CRC Press, Boca Raton

    Google Scholar 

  14. Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl AcadSci USA 88:9828–9832

    CAS  Article  Google Scholar 

  15. Müller M, Irkens-Kiesecker U, Rubinstein B, Taiz L (1996) On the mechanism of hyperacidification in lemon. J Biol Chem 271:1916–1924

    Article  PubMed  Google Scholar 

  16. Nanos GD, Kader A (1993) Low O2-induced changes in pH and energy charge in pear fruit tissue. Postharvest Biol Technol 3:285–291

    CAS  Article  Google Scholar 

  17. Paterson AH, Lander ES, Hewitt JD, Peterson S, Lincoln SE, Tanksley SD (1988) Resolution of quantitative traits into mendelian factors by using a complete linkage map of restriction fragment length polymorphisms. Nature 335:721–726

    CAS  Article  PubMed  Google Scholar 

  18. Pereira da Costa JH, Rodríguez GR, Pratta GR, Picardi LA, Zorzoli R (2013) QTL detection for fruit shelf life and quality traits across segregating populations of tomato. Sci Hortic 156:47–53

    Article  Google Scholar 

  19. Pereira da Costa JH, Rodríguez GR, Pratta GR, Picardi LA, Zorzoli R (2014) Pericarp polypeptides and SRAP markers associated with fruit quality traits in an interspecific tomato backcross. Genet Mol Res 13:2539–2547

    CAS  Article  PubMed  Google Scholar 

  20. Pinheiro SCF, Almeida DPF (2008) Modulation of tomato pericarp firmness through pH and calcium: implications for the texture of fresh-cut fruit. Postharvest Biol Technol 47:119–125

    CAS  Article  Google Scholar 

  21. Rice AC, Pederson CS (1954) Factors influencing growth of Bacillus coagulans in canned tomato juice. II. Acidic constituents of tomato juice and specific organic acids. J Food Sci 19:124–133

    CAS  Article  Google Scholar 

  22. Rocco M, D’Ambrosio C, Arena S, Faurobert M, Scaloni A, Marra M (2006) Proteomic analysis of tomato fruits from two ecotypes during ripening. Proteomics 6:3781–3791

    CAS  Article  PubMed  Google Scholar 

  23. Rodríguez GR, Pratta GR, Zorzoli R, Picardi LA (2006) Recombinant lines obtained from an interspecific cross between Lycopersicon species selected by fruit weight and fruit shelf life. JASHS 131:651–656

    Google Scholar 

  24. Rodríguez GR, Sequin L, Pratta GR, Zorzoli R, Picardi LA (2008) Protein profiling in F1 and F2 generations of two tomato genotypes differing in ripening time. Biol Plant 52:548–552

    Article  Google Scholar 

  25. Rodríguez GR, Pereira da Costa JH, Tomat D, Pratta GR, Zorzoli R, Picardi LA (2011) Pericarp total protein profiles as molecular markers of tomato fruit quality traits in two segregating populations. Sci Hortic 130:60–66

    Article  Google Scholar 

  26. Rodríguez GR, Kim H, van der Knaap E (2013) Mapping of two suppressors of OVATE (sov) loci in tomato. Heredity 111:256–264

    Article  PubMed  PubMed Central  Google Scholar 

  27. Saliba-Colombani V, Causse M, Langlois D, Philouze J, Buret M (2001) Genetic analysis of organoleptic quality in fresh market tomato. 1. Mapping QTLs for physical and chemical traits. Theor App Genet 102:259–272

    CAS  Article  Google Scholar 

  28. Senior AE, Nadanaciva S, Weber J (2002) The molecular mechanism of ATP synthesis by F1F0-ATP synthase. BBA Biochim Biophys Acta 1553:188–211

    CAS  Article  PubMed  Google Scholar 

  29. Shapiro SS, Wilk MB (1965) An analysis of variance test for normality (complete samples). Biometrika 52:591–611

    Article  Google Scholar 

  30. Snedecor G (1964) Métodos Estadísticos. Compañía Editorial

  31. Tanksley SD (1993) Mapping polygenes. Annu Rev Genet 27:205–233

    CAS  Article  PubMed  Google Scholar 

  32. Xu J, Pascual L, Aurand R, Bouchet J, Valot B, Zivy M, Causse M, Faurobert M (2013) An extensive proteome map of tomato (Solanum lycopersicum) fruit pericarp. Proteomics 20:3059–3063

    Google Scholar 

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Acknowledgements

The authors gratefully acknowledge the Tomato Genetics Resource Center (University of California, Davis, CA) for kindly providing seeds of the accession LA722 of Solanum lycopersicum. Thanks to Gabriela Venturi for English edition. This work was supported by Agencia Nacional de Promoción Científica y Tecnológica de Argentina (ANPCyT).

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Correspondence to Gustavo R. Rodríguez.

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J. H. Pereira da Costa and T. A. Vega contributed equally.

Communicated by J-H Liu.

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Pereira da Costa, J.H., Vega, T.A., Pratta, G.R. et al. A 54-kDa polypeptide identified by 2D-PAGE and bulked segregant analysis underlies differences for pH values in tomato fruit. Acta Physiol Plant 39, 78 (2017). https://doi.org/10.1007/s11738-017-2386-9

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Keywords

  • Fruit quality
  • Proteomics
  • SDS–PAGE
  • Solanum lycopersicum
  • Solanum pimpinellifolium