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The novel Solanum tuberosum calcium dependent protein kinase, StCDPK3, is expressed in actively growing organs

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Abstract

Calcium-dependent protein kinases (CDPKs) are key components of calcium regulated signaling cascades in plants. In this work, isoform StCDPK3 from Solanum tuberosum was studied and fully described. StCDPK3 encodes a 63 kDa protein with an N-terminal variable domain (NTV), rich in prolines and glutamines, which presents myristoylation and palmitoylation consensus sites and a PEST sequence indicative of rapid protein degradation. StCDPK3 gene (circa 11 kb) is localized in chromosome 3, shares the eight exons and seven introns structure with other isoforms from subgroup IIa and contains an additional intron in the 5′UTR region. StCDPK3 expression is ubiquitous being transcripts more abundant in early elongating stolons (ES), leaves and roots, however isoform specific antibodies only detected the protein in leaf particulate extracts. The recombinant 6xHis-StCDPK3 is an active kinase that differs in its kinetic parameters and calcium requirements from StCDPK1 and 2 isoforms. In vitro, StCDPK3 undergoes autophosphorylation regardless of the addition of calcium. The StCDPK3 promoter region (circa 1,800 bp) was subcloned by genome walking and fused to GUS. Light and ABRE responsive elements were identified in the promoter region as well as elements associated to expression in roots. StCDPK3 expression was enhanced by ABA while GA decreased it. Potato transgenic lines harboring StCDPK3 promoter∷GUS construct were generated by Agrobacterium tumefaciens mediated plant transformation. Promoter activity was detected in leaves, root tips and branching points, early ES, tuber eyes and developing sprouts indicating that StCDPK3 is expressed in actively growing organs.

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Abbreviations

ABA:

Abscisic acid

CDPK:

Calcium dependent protein kinase

CaM:

Calmodulin

NTV:

N-terminal variable

2-DE:

Two-dimensional gel electrophoresis

PGA:

Polygalacturonic acid

GA3:

Gibberellic acid

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Acknowledgments

We thank Salomé Prat Monguío (CSIC, Madrid, Spain) for the cDNA library from tuberizing stolons. RMU is member of CONICET. CG, VG and FS are fellows from CONICET. This work was supported by grants from CONICET (PIP 00037), University of Buenos Aires (UBACYT X225) and FONCYT-ANPCYT (PICT 2005 #32560). Contributions from DH’s group were supported by the NSF Plant Genome Research Program award no. DBI-0820659 and National Research Initiative grant no. 2008-02806 from the USDA National Institute of Food and Agriculture Iowa State University (ISU).

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Authors and Affiliations

  1. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular “Dr. Hector N. Torres”, Consejo Nacional de Investigaciones Científicas y Técnicas, Vuelta de Obligado 2490 2° piso, C1428ADN, Buenos Aires, Argentina

    Carolina Grandellis, Verónica Giammaria, Magalí Bialer, Franco Santin & Rita M. Ulloa

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

    Verónica Giammaria & Rita M. Ulloa

  3. Plant Biology Major, 253 Horticulture Hall, Iowa State University, Ames, IA, 50011-1100, USA

    Tian Lin & David J. Hannapel

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  1. Carolina Grandellis
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  2. Verónica Giammaria
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  3. Magalí Bialer
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  4. Franco Santin
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  5. Tian Lin
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  7. Rita M. Ulloa
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Correspondence to Rita M. Ulloa.

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Grandellis, C., Giammaria, V., Bialer, M. et al. The novel Solanum tuberosum calcium dependent protein kinase, StCDPK3, is expressed in actively growing organs. Planta 236, 1831–1848 (2012). https://doi.org/10.1007/s00425-012-1732-9

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