Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 99, Issue 2, pp 141–149 | Cite as

CDPK gene expression in somatic embryos of Panax ginseng expressing rolC

  • Konstantin V. Kiselev
  • Anna V. Turlenko
  • Yuri N. Zhuravlev
Original Paper

Abstract

The 2c3 embryogenic culture was obtained as a result of transferring the rolC oncogene from Agrobacterium rhizogenes to the callus cells of Panax ginseng. Calcium-dependent protein kinases (CDPKs) are known to play a role in the development of somatic embryos in the 2c3 cell culture. Ten CDPK genes with altered expressions in the 2c3 embryogenic cell culture have previously been described. In this study, the importance of the ginseng CDPK gene for stimulation of somatic embryogenesis was investigated. Frequency analysis of RT-PCR products and real-time PCR were used to analyze CDPK gene expression in the 2c3 callus at different stages of somatic embryo development. Our results suggest that members of the PgCDPK2d subfamily (PgCDPK2d, PgCDPK2ds, and PgCDPK2dL) play a role in the initialization and development of somatic embryos. It was also found that the kinase domain of these genes was subjected to insertion and deletion modifications. The observed transcriptional and post-transcriptional modifications (alternative splicing, RNA editing or nonsense-mediated mRNA decay) of the PgCDPK2d genes could contribute to the formation of somatic embryos initiated by the rolC oncogene.

Keywords

Agrobacterium rhizogenes CDPK Genes rol Panax ginseng rolC Somatic embryogenesis 

Abbreviations

CDPK

Calcium-dependent protein kinases

RT-PCR

Reverse transcription PCR

Notes

Acknowledgments

This work was supported by grants from the Russian Foundation for Basic Research (06-04-48149-a), the President of Russia (MК-714.2008.4), the Far East Division of the Russian Academy of Sciences (06-III-A-06-146), and the Russian Science Support Foundation.

References

  1. Asano T, Tanaka N, Yang G, Hayashi N, Komatsu S (2005) Genome-wide identification of the rice calcium-dependent protein kinase and its closely related kinase gene families: comprehensive analysis of the CDPKs gene family in rice. Plant Cell Physiol 46:356–366PubMedCrossRefGoogle Scholar
  2. Baker KE, Parker R (2004) Nonsense-mediated mRNA decay: terminating erroneous gene expression. Curr Opin Cell Biol 16:293–299PubMedCrossRefGoogle Scholar
  3. Benfey PN, Chua NH (1990) The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science 250:959–966PubMedCrossRefGoogle Scholar
  4. Bonhomme V, Laurain Mattar D, Fliniaux MA (2000) Effects of the rolC gene on hairy root: Induction development and tropane alkaloid production by Atropa belladonna. J Nat Prod 63:1249–1252PubMedCrossRefGoogle Scholar
  5. Bulgakov VP, Veselova MV, Tchernoded GK, Kiselev KV, Fedoreyev SA, Zhuravlev YN (2005) Inhibitory effect of the Agrobacterium rhizogenes rolC gene on rabdosiin and rosmarinic acid production in Eritrichium sericeum and Lithospermum erythrorhizon transformed cell cultures. Planta 221:471–478PubMedCrossRefGoogle Scholar
  6. Chang WC, Hsing Y (1980) In vitro flowering of embryoids derived from mature root callus of ginseng (Panax ginseng). Nature 284:341–342CrossRefGoogle Scholar
  7. Cheng SH, Willmann MR, Chen HC, Sheen J (2002) Calcium signaling through protein kinases. The Arabidopsis calcium-dependent protein kinase gene family. Plant Physiol 129:469–485PubMedCrossRefGoogle Scholar
  8. Costantino P, Capone I, Cardarelli M, De Paolis A, Mauro ML, Trovato M (1994) Bacterial plant oncogenes—the rol genes saga. Genetica 94:203–211PubMedCrossRefGoogle Scholar
  9. Delbarre A, Muller P, Imhoff V, Barbier-Brygoo H, Maurel C, Leblanc N, Perrot-Rechenmann C, Guern J (1994) The rolB gene of Agrobacterium rhizogenes does not increase the auxin sensitivity of tobacco protoplasts by modifying the intracellular auxin concentration. Plant Physiol 105:563–569PubMedGoogle Scholar
  10. Dubrovina AS, Kiselev KV, Veselova MV, Isaeva GA, Fedoreyev SA, Zhuravlev YN (2009) Enhanced resveratrol accumulation in rolB transgenic cultures of Vitis amurensis correlates with unusual changes in CDPK gene expression. J Plant Physiol 166:1194–1206PubMedCrossRefGoogle Scholar
  11. Estruch JJ, Chriqui D, Grossmann K, Schell J, Spena A (1991) The plant oncogene rolC is responsible for the release of cytokinins from glucoside conjugates. EMBO J 10:2889–2895PubMedGoogle Scholar
  12. Faiss M, Strnad M, Redig P, Dolezal K, Hanus J, VanOnckelen H, Schmulling T (1996) Chemically induced expression of the rolC encoded β-glucosidase in transgenic tobacco plants and analysis of cytokinin metabolism: rolC does not hydrolyze endogenous cytokinin glucosides in planta. Plant J 10:33–46CrossRefGoogle Scholar
  13. Giulietti A, Overbergh L, Valckx D, Decallonne B, Bouillon R, Mathieu C (2001) An overview of real-time quantitative PCR: applications to quantify cytokine gene expression. Methods 25:386–401PubMedCrossRefGoogle Scholar
  14. Gorpenchenko TY, Kiselev KV, Bulgakov VP, Tchernoded GK, Bragina EA, Khodakovskaya MV, Koren OG, Batygina TB, Zhuravlev YN (2006) The Agrobacterium rhizogenes rolC-gene-induced somatic embryogenesis and shoot organogenesis in Panax ginseng transformed calluses. Planta 223:457–467PubMedCrossRefGoogle Scholar
  15. Harper JF, Harmon A (2005) Plants, symbiosis and parasites: a calcium signaling connection. Nat Rev Mol Cell Biol 6:555–566PubMedCrossRefGoogle Scholar
  16. Kiselev KV, Tchernoded GK (2009) Somatic embryogenesis in the Panax ginseng cell culture induced by the rolC oncogene is associated with increased expression of WUS and SERK genes. Russ J Genet 45:445–452CrossRefGoogle Scholar
  17. Kiselev KV, Kusaykin MI, Dubrovina AS, Bezverbny DA, Zvyagintseva TN, Bulgakov VP (2006) The rolC gene induces expression of a pathogenesis-related beta-1, 3-glucanase in transformed ginseng cells. Phytochemistry 67:2225–2231PubMedCrossRefGoogle Scholar
  18. Kiselev KV, Dubrovina AS, Veselova MV, Bulgakov VP, Fedoreyev SA, Zhuravlev YN (2007) The rolB gene-induced overproduction of resveratrol in Vitis amurensis transformed cells. J Biotechnol 128:681–692PubMedCrossRefGoogle Scholar
  19. Kiselev KV, Gorpenchenko TY, Tchernoded GK, Dubrovina AS, Grishchenko OV, Bulgakov VP, Zhuravlev YN (2008) Calcium-dependent mechanism of somatic embryogenesis in Panax ginseng cell cultures expressing the rolC oncogene. Mol Biol 42:243–252CrossRefGoogle Scholar
  20. Lecourieux D, Ranjeva R, Pugin A (2006) Calcium in plant defence-signalling pathways. New Phytol 171:249–269PubMedCrossRefGoogle Scholar
  21. Ludwig AA, Romeis T, Jones JD (2004) CDPK-mediated signalling pathways: specificity and cross-talk. J Exp Bot 55:181–188PubMedCrossRefGoogle Scholar
  22. Nagy E, Maquat LE (1998) A rule for termination-codon position within intron-containing genes: when nonsense affects RNA abundance. Trends Biochem Sci 23:198–199PubMedCrossRefGoogle Scholar
  23. Nilsson O, Olsson O (1997) Getting to the root: the role of the Agrobacterium rhizogenes rol genes in the formation of hairy roots. Physiol Plant 100:463–473CrossRefGoogle Scholar
  24. Nilsson O, Moritz T, Sundberg B, Sandberg G, Olsson O (1996) Expression of the Agrobacterium rhizogenes rolC gene in a deciduous forest tree alters growth and development and leads to stem fasciation. Plant Physiol 112:493–502PubMedGoogle Scholar
  25. Nishiyama R, Mizuno H, Okada S, Yamaguchi T, Takenaka M, Fukuzawa H, Ohyama K (1999) Two mRNA species encoding calcium-dependent protein kinases are differentially expressed in sexual organs of Marchantia polymorpha through alternative splicing. Plant Cell Physiol 40:205–212PubMedGoogle Scholar
  26. Park SH, Choi J, Kang JI, Choi SY, Hwang SB, Kim JP, Ahn BY (2006) Attenuated expression of interferon-induced protein kinase PKR in a simian cell devoid of type I interferons. Mol Cells 21:21–28PubMedGoogle Scholar
  27. Schmulling T, Shell J, Spena A (1988) Single genes from Agrobacterium rhizogenes influence plant development. EMBO J 7:2621–2629PubMedGoogle Scholar
  28. Schmulling T, Fladung M, Grossman K, Schell J (1993) Hormonal content and sensitivity of transgenic tobacco and potato plants expressing single rol genes of Agrobacterium rhizogenes T-DNA. Plant J 3:371–382Google Scholar
  29. Spena A, Schmulling T, Koncz C, Schell JS (1987) Independent and synergistic activity of rolA, B and C loci in stimulating abnormal growth in plants. EMBO J 6:3891–3899PubMedGoogle Scholar
  30. Wei YH, Fu GL, Hu HR, Lin G, Yang JC, Guo JH, Zhu QQ, Yu L (2007) Isolation and characterization of mouse testis specific ser/thr kinase 5 possessing four alternatively spliced variants. J Biochem Mol Biol 40:749–756PubMedGoogle Scholar
  31. Xiong LZ, Yang YN (2003) Disease resistance and abiotic stress tolerance in rice are inversely modulated by an abscisic acid-inducible mitogen-activated protein kinase. Plant Cell 15:745–759PubMedCrossRefGoogle Scholar
  32. Yue PY, Mak NK, Cheng YK, Leung KW, Ng TB, Fan DT, Yeung HW, Wong RN (2007) Pharmacogenomics and the Yin/Yang actions of ginseng: anti-tumor, angiomodulating and steroid-like activities of ginsenosides. Chin Med 6:1–21Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Konstantin V. Kiselev
    • 1
  • Anna V. Turlenko
    • 1
  • Yuri N. Zhuravlev
    • 1
  1. 1.Laboratory of Biotechnology, Institute of Biology and Soil ScienceFar East Branch of Russian Academy of SciencesVladivostokRussia

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