Skip to main content
SpringerLink
Log in

Overexpression of glycine-rich RNA-binding protein in tomato renders fruits with higher protein content after cold storage

  • Original paper
  • Published:
Biologia Plantarum

Abstract

Glycine-rich RNA-binding proteins (GR-RBPs) are involved in RNA processing and also some of them are output signals of the circadian clock. In tomato, one GR-RBP gene family (LeGRP1) is composed by three highly homologous genes (LeGRP1a-c); each one rendering three transcriptional products: the un-spliced pre-RNA (preLegrp1a-c), the mature mRNA (mLegrp1a-c) and the alternatively spliced mRNA (asLegrp1a-c). To get insight into their regulation and impact on RNA metabolism in fruits, Solanum lycopersicum cv. Micro-Tom was transformed with preLeGRP1a fused to the polygalacturonase promoter, which drives expression to fruits from the mature green stage. Our results demonstrated a complex positive regulation of LeGRPs, in which LeGRP1a overexpression led to the induction of the others LeGRP1 members. Even though the LeGRP1 transcription and the content of three LeGRPs proteins were affected, the overall LeGRP protein circadian rhythm profile was similar in transgenic and wild type (WT) fruits. However, when the fruits were kept at a chilling temperature after harvest, total protein content was significantly higher in transgenic than in WT fruits, and the content of some free amino acids was modified. The results obtained suggest a probable role of LeGRP1s: structural rearrangements and/or stabilization of mRNA to allow efficient processing of fruits under cold conditions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

CI:

chilling injury

CSD:

cold shock domain

DAA:

days after anthesis

GR-RBPs:

glycine-rich RNA-binding proteins

IG:

immature green

MG:

mature green

PG:

polygalacturonase

PITC:

phenylisothiocyanate

RT-qPCR:

reverse

transcription:

quantitative polymerase chain reaction

RBP:

RNA binding protein

RR:

red ripe

RRM:

RNA recognition motif

TEA:

triethylamine

WT:

wild-type

Y:

green yellowish

References

  • Ambrosone, A., Batelli, G., Nurcato, R., Aurilia, V., Punzo, P., Bangarusamy, D K., Ruberti, I., Sassi, M., Leone A., Costa, A., Grillo, S.: The Arabidopsis RNA-binding protein AtRGGA regulates tolerance to salt and drought stress. - Plant Physiol. 168: 292–306, 2015.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • An, G., Ebert, P.R., Mitra, A. Ha, S.B.: Binary Vectors in Plant Molecular Biology. - Kluwer Academic Publishers, Dordrect 1988.

    Google Scholar 

  • Ayarpadikannan, S., Chung, E.S., So, H.A., Kim, K.M., Schraufnagle, K.R., Lee, J.H.: Overexpression of SaRBP1 enhances tolerance of Arabidopsis to salt stress. - Plant Cell Tissue Organ Cult. 118: 327–338, 2014.

    Article  CAS  Google Scholar 

  • Borsani, J., Budde, C.O., Porrini, L., Lauxmann, M.A., Lombardo, V.A., Murray, R., Andreo, C.S., Drincovich, M.F., Lara, M.V.: Carbon metabolism of peach fruit after harvest: changes in enzymes involved in organic acid and sugar level modifications. - J. exp. Bot 60: 1823–1837, 2009.

    Article  PubMed  CAS  Google Scholar 

  • Bradford, M.M.: 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, 1976.

    Article  PubMed  CAS  Google Scholar 

  • Burnette, W.N.: "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. - Anal Biochem. 112: 195–203, 1981.

    Article  PubMed  CAS  Google Scholar 

  • Cook, K.B., Kazan, H., Suberi, K., Morris, Q., Hughes, T.R.: RBPDB: a database of RNA-binding specificities. - Nucl. Acids Res. 39: 301–308, 2011.

    Article  CAS  Google Scholar 

  • Dhar, P., Tayade, A.B., Kumar, J., Chaurasia, O.P., Srivastava, R.B., Singh, S.B.: Nutritional profile of phytococktail from trans-Himalayan plants. - PLoS One 8: e83008, 2013.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Fahnenstich, H., Saigo, M., Niessen, M., Zanor, M.I., Andreo, C.S., Fernie, A., Drincovich, M.F., Flügge, U.I., Maurino, V.G.: Alteration of organic acid metabolism in Arabidopsis overexpressing the maize C4 NADP-malic enzyme causes accelerated senescence during extended darkness. - Plant Physiol. 145: 640–652, 2007.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Fernandez, A.I., Viron, N., Alhagdow, M., Karimi, M., Jones, M., Amsellem, Z., Sicard, A., Czerednik, A., Angenent, G., Grierson, D., May, S., Seymour, G., Eshed, Y., Lemaire- Chamley, M., Rothan, C., Hilson, P.: Flexible tools for gene expression and silencing in tomato. - Plant Physiol. 151: 1729–1740, 2009.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Jackman, R.L., Yada, R.Y., Marangoni, A., Parkinz, K.L., Stanley, D.W.: Chilling injury. A review of quality aspects. - J. Food Qual. 11: 253–278, 1988.

    Article  Google Scholar 

  • Jung, H.J., Park, S.J., Kang, H.: Regulation of RNA metabolism in plant development and stress responses. - J. Plant Biol. 56: 123–129, 2013.

    Article  CAS  Google Scholar 

  • Kang, H., Park, S.J., Kwak, K.J.: Plant RNA chaperones in stress response. - Trends Plant Sci. 18: 100–106, 2013.

    Article  PubMed  CAS  Google Scholar 

  • Khan, F., Daniëls, M.A., Folkers, G.E., Boelens, R., Saqlan Naqvi, S.M., Van Ingen, H.: Structural basis of nucleic acid binding by Nicotiana tabacum glycine-rich RNA-binding protein: implications for its RNA chaperone function. - Nucl. Acids Res. 42: 8705–8718, 2014.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Kim, J.S., Jung, H.J., Lee, H.J., Kim, K.A., Goh, C.H., Woo, Y., Oh, S.H., Han, Y.S., Kang, H.: Glycine-rich RNAbinding protein7 affects abiotic stress responses by regulating stomata opening and closing in Arabidopsis thaliana. - Plant J. 55: 455–466, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Kim, J.S., Park, S.J., Kwak, K.J., Kim, Y.O., Kim, J.Y., Song, J., Jang, B., Jung, C.-H., Kang, H.: Cold shock domain proteins and glycine-rich RNA-binding proteins from Arabidopsis thaliana can promote the cold adaptation process in E. coli. - Nucl. Acids Res. 35, 506–516, 2007b.

    Article  PubMed  CAS  Google Scholar 

  • Kim, J.Y., Kim, W.Y., Kwak, K.J., Oh, S.H., Han, Y.S., Kang, H.: Glycine-rich RNA-binding proteins are functionally conserved in Arabidopsis thaliana and Oryza sativa during cold adaptation process. - J. exp. Bot. 61: 2317–2325, 2010.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Kim, J.Y., Park, S.J., Jang, B., Jung, C.-H., Ahn, S.J., Goh, C.-H., Cho, K., Han, O., Kang, H.: Functional characterrization of a glycine-rich RNA-binding protein 2 in Arabidopsis thaliana under abiotic stress conditions. - Plant J. 50: 439–451, 2007a.

    Article  PubMed  CAS  Google Scholar 

  • Kim, M.K., Jung, H.J., Kim, D.H., Kang, H.: Characterization of glycine-rich RNA-binding proteins in Brassica napus under stress conditions. - Physiol. Plant 146: 297–307, 2012.

    Article  PubMed  CAS  Google Scholar 

  • Kwak, K.J., Kim, H.-S., Jang, H.Y., Kang, H., Ahn, S.-J.: Diverse roles of glycine-rich RNA-binding protein 7 in the response of camelina (Camelina sativa) to abiotic stress. - Acta Physiol. Plant. 38: 129, 2016.

    Article  CAS  Google Scholar 

  • Kwak, K.J., Park, S.J., Han, J.H., Kim, M.K., Oh, S.H., Han, Y.S., Kang, H.: Structural determinants crucial to the RNA chaperone activity of glycine-rich RNA-binding proteins 4 and 7 in Arabidopsis thaliana during the cold adaptation process. - J. exp. Bot. 62: 4003–4011, 2011.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. - Nature 227: 680–685, 1970.

    Article  PubMed  CAS  Google Scholar 

  • Lana, M.M., Tijskens, L.M.M., Van Kooten, O.: Effects of storage temperature and fruit ripening on firmness of fresh cut tomatoes. - Postharvest Biol. Technol. 35: 87–89, 2005.

    Article  Google Scholar 

  • Lau, J.M., Cooper, N.G., Robinson, D.L., Korban, S.S.: Sequence and in silico characterization of the tomato polygalacturonase (PG) promoter and terminator regions. - Plant mol. Biol. Rep. 27: 250–256, 2009.

    Article  CAS  Google Scholar 

  • López Camelo, A.F.: Storage. - In: Manual for thePreparation and Sale of Fruits and Vegetables from Field to Market. Pp. 47–74. FAO, Rome 2004.

    Google Scholar 

  • Lorković, Z.J.: Role of plant RNA-binding proteins in development. stress response and genome organization. - Trends Plant Sci. 14: 229–236, 2009.

    Article  PubMed  CAS  Google Scholar 

  • Müller, G.L., Budde, C.O., Lauxmann, M.A., Triassi, A., Andreo, C.S., Drincovich, M.F., Lara, M.V.: Expression profile of transcripts encoding cell wall remodelling proteins in tomato fruit cv. Micro-Tom subjected to 15 °C storage. - Funct. Plant Biol. 40: 449–458, 2013.

    Article  CAS  Google Scholar 

  • Müller, G.L., Triassi, A., Alvarez, C., Falcone Ferreyra, M.L., Andreo, C.S., Lara, M.V., Drincovich, M.F.: Circadian oscillation and development-dependent expression of glycine-rich RNA binding proteins in tomato fruits. - Funct. Plant Biol. 41: 411–423, 2014.

    Article  CAS  Google Scholar 

  • Nagata, M., Yamashita, I.: Simple method for simultaneous determination of chlorophyll and carotenoids in tomato fruit. - J. jap. Soc Food Sci 39: 925–928, 1992.

    CAS  Google Scholar 

  • Park, E.J., Jeknić, Z., Pino, M.T., Murata, N., Chen, T.H.: Glycinebetaine accumulation is more effective in chloroplasts than in the cytosol for protecting transgenic tomato plants against abiotic stress. - Plant Cell Environ. 8: 994–1005, 2007.

    Article  CAS  Google Scholar 

  • Phadtare, S.: Unwinding activity of cold shock proteins and RNA metabolism. - RNA Biol. 8: 394–397, 2011.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pratelli, R. Pilot, G.: Regulation of amino acid metabolic enzymes and transporters in plants. - J. exp. Bot. 65: 5535–5556, 2014.

    Article  PubMed  CAS  Google Scholar 

  • Sachetto-Martins, G., Franco, L.O., De Oliveira, D.E.: Plant glycine-rich proteins: a family or just proteins with a common motif? - Biochim. biophys. Acta 1492: 1–14, 2000.

    Article  PubMed  CAS  Google Scholar 

  • Sahi, C., Agarwal, M., Singh, A., Grover, A.: Molecular characterization of a novel isoform of rice (Oryza sativa L.) glycine rich-RNA binding protein and evidence for its involvement in high temperature stress response. - Plant Sci. 173: 144–155, 2007.

    Article  CAS  Google Scholar 

  • Sakamoto, A., Murata, N.: The role of glycine betaine in the protection of plants from stress: clues from transgenic plants. - Plant Cell Environ. 25: 163–171, 2002.

    Article  PubMed  CAS  Google Scholar 

  • Sasaki, K., Imai, R.: Pleiotropic roles of cold shock domain proteins in plants. - Front. Plant Sci. 2: 116, 2012.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Schöning, J.C., Streitner, C., Meyer, I.M., Gao, Y., Staiger, D.: Reciprocal regulation of glycine-rich RNA-binding proteins via an interlocked feedback loop coupling alternative splicing to nonsense-mediated decay in Arabidopsis reciprocal regulation of glycine-rich RNA-binding proteins via an interlocked feedback loop. - Nucl. Acids Res. 36: 6977–6987, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Schöning, J.C., Streitner, C., Page, D.R., Hennig, S., Uchida, K., Wolf, E., Furuya, M., Staiger, D.: Autoregulation of the circadian slave oscillator component AtGRP7 and regulation of its targetstargets is impaired by a single RNA recognition motif point mutation. - Plant J. 5: 1119–1130, 2007.

    Article  CAS  Google Scholar 

  • Staiger, D., Zecca, L., Wieczorek Kirk, D.A., Apel, K., Eckstein, L.: The circadian clock regulated RNA-binding protein AtGRP7 autoregulates its expression by influencing alternative splicing of its own pre-mRNA. - Plant J. 33: 361–371, 2003.

    Article  PubMed  CAS  Google Scholar 

  • Tan, Y., Qin, Y., Li, Y., Li, M., Ma, F.: Overexpression of MpGR-RBP1, a glycine-rich RNA-binding protein gene from Malus prunifolia (Willd.) Borkh, confers salt stress tolerance and protects against oxidative stress in Arabidopsis. - Plant Cell Tissue Organ Cult. 119: 635–646, 2014.

    Article  CAS  Google Scholar 

  • Zhang, B., Tieman, D.M., Jiao, C., Xu, Y., Chen, K., Fe, Z., Giovannoni, J.J., Klee, H.J.: Chilling-induced tomato flavor loss is associated with altered volatile synthesis and transient changes in DNA methylation. - Proc. nat. Acad. Sci. USA 113: 12580–12585, 2016.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centro de Estudios Fotosintéticos y Bioquímicos, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, 2000, Argentina

    G. M. Ruggieri, A. Triassi, C. E. Alvarez, A. Gola, J. Wiggenhauser, M. V. Lara, M. F. Drincovich & G. L. Müller

  2. Estación Experimental San Pedro, Instituto Nacional de Tecnología Agropecuaria, San Pedro, 2930, Argentina

    C. O. Budde

Authors
  1. G. M. Ruggieri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Triassi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. E. Alvarez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. Gola
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. Wiggenhauser
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C. O. Budde
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M. V. Lara
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. M. F. Drincovich
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. G. L. Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. L. Müller.

Additional information

Acknowledgments: This work was funded by the Argentine National Research Council and the National Agency for Science Promotion and Technology (PICT 2012 Nº19). The first two authors contributed equally to this work.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ruggieri, G.M., Triassi, A., Alvarez, C.E. et al. Overexpression of glycine-rich RNA-binding protein in tomato renders fruits with higher protein content after cold storage. Biol Plant 62, 501–510 (2018). https://doi.org/10.1007/s10535-018-0794-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10535-018-0794-3

Additional key words

Search

Navigation