The success of nuclear transplantation with mammalian oocytes depends critically on the potential of oocytes activation, which mainly caused to prevent the re-accumulation of maturation promoting factor (MPF). This study was conducted to compare the effect of combined treatment of lonomycin with a Hl-histone kinase inhibitor (dimethylaminopurine, DMAP) or cdc2 kinase inhibitor (sodium pyrophosphate, SPP) on activation of bovine oocytes. In vitro matured bovine oocytes with the first polar body (PB) and dense cytoplasm were assigned to 3 experimental groups. For activation treatment, oocytcs were exposed to 5 M lonomycin for 5 min (Group 1), and followed by 1.9 mM dimethylaminopurine (DMAP) for 3 h (Group 2) or followed by 2 mM sodium pyrophosphate (SPP) for 3 h (Group 3). The activation effects in the three treatments and the control group (untreated) were judged by the extrusion of the second PB and formation of a pronucleus (PN). Differences among groups were analysed using one-way ANOVA after arc-sine transformation of proportional data. All three treatments led to high activation rates (90% to 95%), with significant difference from the control. However, the extrusion of the second PB and the rate of PN formation differed remarkably among treatments. In Group I and 3, about 95% of the oocytes had extruded the second polar body, but one PN had formed in a higher proportion of oocytes in Group 3 than in Group 1 (90% vs. 5%). In experiment 2, the rates of cleavage and development into blastocysts in Group 1 were significantly lower than those of Group 2 and 3 (8.7% and 0% vs. 50.5% and 11.6%, and 44.6% and 7.2%, respectively, P<0.05). In experiment 3, ~80% of parthenotes in Group 1 were developed with haploid chromosomal sets. However, when ionomycin was followed immediately by DMAP (Group 2). only 20% of parthenotes were haploid. In Group 3, combined treatment with ionomycin and SPP, the appearance of abnormal chromosomal tracts was significantly (P〈0.05) reduced and the proportion of haploid parthenotes was increased to 85% (17/20) than in Group 2. These results demonstrate that SPP acted as a cdc2 kinase inhibitor and formed the haploidy in oocyte activation. Thus, the present study suggests that cdc2 kinase inhibitor, such as sodium pyrophosphate, may have an effective role in oocyte activation for the production of cloned embryos/animals by nuclear transplantation.

The occurrence of major pest infestation was compared between conventional tea plantation and organic tea plantation at Sulloc tea garden in Dosun-dong, Seogwipo-si, Jeju-do from 2002 to 2009. Tetranychus kanzawai was observed a lot in the second year, but it waned from late June. There was not much difference between conventional farming and organic farming in terms of pest density. Empoasca onukii was infested in the second year of organic farming compared with conventional farming, which highlighted the fact that second year of organic farming requires a special care. Scirotothrips dorsalis was highly dense in the second and third year of conventional farming, but its occurrence was lowered when the farming technique was shifted to organic farming. The number of Homona magnanima peaked 4 times each year. In 2008, the first year of organic farming, saw high occurrence of 771.2 per trap per year. In 2009, the second year, the population per trap dropped to 80, showing a great variance depending on year. The occurrence of Caloptilia theivora peaked 5 times annually. In 2008, the first year of organic farming, an average of 2,779 pests per trap was found, and in the following year, 4,143 pests were observed. It showed that density rose in organic growing period.