RESUMO
The embryogenic cell culture 2c3 was previously obtained by the transfer of the rolC gene from Agrobacterium rhizogenes into ginseng callus cells. It was found by us that the expression of SERK and WUS genes in the embryogenic culture 2c3 is increased. These genes are known to be responsible for the initial stimulus to the development of embryogenesis in plants. Taking into consideration earlier data, we suppose that the development of somatic embryos in the ginseng cell culture 2c3 is associated with changes in the work of the calcium signaling system and with the activation of expression of SERK and WUS genes.
Assuntos
Regulação da Expressão Gênica de Plantas , Genes Bacterianos , Proteínas de Homeodomínio/biossíntese , Panax/embriologia , Proteínas de Plantas/biossíntese , Proteínas Quinases/biossíntese , Rhizobium , Proteínas de Homeodomínio/genética , Panax/genética , Proteínas de Plantas/genética , Proteínas Quinases/genéticaRESUMO
It was shown earlier, that ginseng embryogenic cell culture 2c3 was obtained as a result of callus cells transformation with the Agrobacterium rhizogenes rolC oncogene. In the present report we determine that inhibitors of Ca2+-channels (LaCl3, verapamil, niflumic acid) certainly lowered the quantity of somatic embryos in the 2c3 cell culture. This is the evidence of the influence of calcium-dependent signal system on plant embryogenesis. Protein kinases inhibitors W7 and H7 also caused the lowering of somatic embryos quantity in the 2c3 cell culture. We analysed changes of CDPK genes expression in embryogenic 2c3 cell culture. Total expression decreased 1.2-1.5 times comparing with the control callus culture. CDPK expression in the 2c3 embryogenic culture lowered by the inhibition of expression of the gene subfamilies PgCDPK1 (PgCDPK1a and PgCDPK1b) and PgCDPK3 (PgCDPK3a). At the same time, expression of PgCDPK2 gene subfamily (PgCDPK2b and PgCDPK2d) was increased. We suppose that genes of PgCDPK2 subfamily might be responsible for the embryogenesis initiation in the 2c3 ginseng cell culture. It was shown for the first time that the rolC gene and the process of embryogenesis could change expression of particular forms of CDPK genes.
Assuntos
Proteínas de Ligação ao Cálcio/biossíntese , Genes Bacterianos , Panax/embriologia , Proteínas de Plantas/biossíntese , Proteínas Quinases/biossíntese , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Sinalização do Cálcio/genética , Proteínas de Ligação ao Cálcio/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/genética , Genes Bacterianos/genética , Panax/citologia , Panax/genética , Proteínas de Plantas/genética , Tumores de Planta/genética , Inibidores de Proteínas Quinases/farmacologia , Proteínas Quinases/genética , Rhizobium/genéticaRESUMO
Expression of the Agrobacterium rhizogenes rolC gene in Panax ginseng callus cells results in formation of tumors that are capable to form roots. The selection of non-root forming tumor clusters yielded the embryogenic 2c3 callus line, which formed somatic embryos and shoots independently of external growth factors. Although the 2c3 somatic embryos developed through a typical embryogenesis process, they terminated prematurely and repeatedly formed adventitious shoot meristems and embryo-like structures. A part of the shoots and somatic embryos formed enlarged and fasciated meristems. This is the first indication of the rolC gene embryogenic effect and, to our knowledge, the first indication that a single gene of non-plant origin can induce somatic embryogenesis in plants.