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Transcript profiling reveals that cysteine protease inhibitors are up-regulated in tuber sprouts after extended darkness

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Abstract

Potato (Solanum tuberosum L.) tubers are an excellent staple food due to its high nutritional value. When the tuber reaches physiological competence, sprouting proceeds accompanied by changes at mRNA and protein levels. Potato tubers become a source of carbon and energy until sprouts are capable of independent growth. Transcript profiling of sprouts grown under continuous light or dark conditions was performed using the TIGR 10K EST Solanaceae microarray. The profiles analyzed show a core of highly expressed transcripts that are associated to the reactivation of growth. Under light conditions, the photosynthetic machinery was fully activated; the highest up-regulation was observed for the Rubisco activase (RCA), the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the Photosystem II 22 kDa protein (CP22) genes, among others. On the other hand, sprouts exposed to continuous darkness elongate longer, and after extended darkness, synthesis of chloroplast components was repressed, the expression of proteases was reduced while genes encoding cysteine protease inhibitors (CPIs) and metallocarboxypeptidase inhibitors (MPIs) were strongly induced. Northern blot and RT-PCR analysis confirmed that MPI levels correlated with the length of the dark period; however, CPI expression was strong only after longer periods of darkness, suggesting a feedback loop (regulation mechanism) in response to dark-induced senescence. Prevention of cysteine protease activity in etiolated sprouts exposed to extended darkness could delay senescence until they emerge to light.

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Acknowledgments

This work was supported by grants from Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), University of Buenos Aires (UBA), and Fondo para la Promocion de Ciencia y Tecnologia (FONCYT-ANPCYT). It was performed in cooperation with the Institute of Genomic Research, Potato Functional Genomics Project, Expression profiling Project Application round 9. RMU is a member of CONICET. CG, VG, IC, SB, EF, and FS are fellows from CONICET.

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Author notes

  1. Carolina Grandellis

    Present address: Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas (IBR-CONICET) Ocampo y Esmeralda, Rosario, Argentina

  2. Ignacio Cerrudo

    Present address: Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura (IFEVA), Universidad de Buenos Aires, and Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín, Buenos Aires, Argentina

Authors and Affiliations

  1. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI), Consejo Nacional de Investigaciones Científicas y Técnicas, Vuelta de Obligado 2490 2nd piso, C1428ADN, Buenos Aires, Argentina

    Carolina Grandellis, Veronica Giammaria, Elisa Fantino, Ignacio Cerrudo, Sandra Bachmann, Franco Santin & Rita M. Ulloa

  2. Consejo Nacional de Investigaciones Científicas y Técnicas, Av. San Martín 4453, C1417DSE, Buenos Aires, Argentina

    Ignacio Cerrudo

  3. Instituto de Biotecnología (IB), Universidad Nacional de Hurlingham, Av. Vergara 2222, Villa Tesei, Hurlingham, B1688GEZ, Buenos Aires, Argentina

    Ignacio Cerrudo

  4. Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina

    Rita M. Ulloa

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  1. Carolina Grandellis
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Correspondence to Rita M. Ulloa.

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Carolina Grandellis and Veronica Giammaria contributed equally to this work.

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ESM Fig. 1

Pictures of tuber sprouts grown for 10 days (a, b, f) or 35 days (c, e) under continuous dark (a, d, e) or light (b, c) conditions. In (e) and (f), 5 μM GA3 was added to the media. Scale bar indicates 10 mm. d Starch content was evaluated in tuber slices using 2-fold serial dilutions of LUGOL solution. (TIF 799 kb)

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Grandellis, C., Giammaria, V., Fantino, E. et al. Transcript profiling reveals that cysteine protease inhibitors are up-regulated in tuber sprouts after extended darkness. Funct Integr Genomics 16, 399–418 (2016). https://doi.org/10.1007/s10142-016-0492-1

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