CDPK gene expression in somatic embryos of Panax ginseng expressing rolC
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.
KeywordsAgrobacterium rhizogenes CDPK Genes rol Panax ginseng rolC Somatic embryogenesis
Calcium-dependent protein kinases
Reverse transcription PCR
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.
- 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
- 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
- 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
- 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
- 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
- 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