[Somatic embryogenesis in the Panax ginseng cell culture induced by the rolC oncogene is associated with increased expression of WUS and SERK genes].

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.

[Calcium-dependent mechanism of somatic embryogenesis in oncogene rolC expressing cell cultures of Panax ginseng].

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.

[Chromosomal variability of ginseng cells transformed with plant oncogene rolC]. / Khromosomnaia variabel'nost' kletok zhen'shenia, transformirovannykh rastitel'nym onkogenom rolC.

Chromosome numbers were was studied in ginseng cell line 1c transformed with Agrobacterium rhizogenes strain A4, which carried plasmid pRiA4, and with A. tumefaciens strain GV3101, which carried vector pPCV002-35S rolC. As compared with the nontransformed cell line 1c, tumor cell cultures 1c-A4 and 1c-rolC and the tissues of rolC teratoma (excluding leaves) displayed higher polyploidy and aneuploidy. The 1c-A4 and 1c-rolC hairy-root cultures also had aneuploid and polyploid cells, but the chromosome variation was lower than in tumor cells or the initial culture 1c. Generally, an increase of chromosome variation in cultivated cells was the main effect of the integration of several oncogenes, which were in the A. rhizogenes A4 T-DNA, or of the individual rolC gene in the ginseng genome. Another effect consisted in stabilization of the chromosome number in some differentiated transgenic tissues. Possible reasons for this effect are discussed.