This study investigated a suitable method that could be applied for Asian chain fern [Woodwardia japonica (L. f.) Sm.] to propagate gametophytes and promote sporophyte formation. The gametophytes used in all experiments were obtained from germinated spores in vitro and were subcultured at 8-week intervals. The most appropriate media for gametophyte propagation was identified by culturing 300 ㎎ of gametophyte in Murashige and Skoog (MS) basal medium (1/8, 1/4, 1/2, 1, 2), and Knop medium for 8 weeks. As a result, fresh weight of the gametophyte was increased by 56.7-fold on MS medium. Moreover, antheridium formation as well as gametophyte growth was improved on MS medium, especially. To improve the sporophyte formation ex vitro, 1.0 g of gametophyte was ground with distilled water and spread on eight combinations onto four different culture mediums, such as bed soil, peat moss, perlite and decomposed granite. Then generation and growth of sporophytes were investigated after cultivation for 10 weeks. As a result of this experiment, peat moss had a promotive effect of sporophyte formation at single-use and mixed culture soils. In particular, a mixture of bed soil, peat moss and perlite in a 1:1:1 ratio (v/v/v) led to the accelerated formation (782.5 ea/pot) and the frond growth of sporophytes. This included increases in length and width of fronds. However, promotive effect of gametophyte growth and sporophyte formation was not found at single-use and treatment with high ratio of bed soil.
To overcome the risk of the ovarian hyperstimulation syndrome (OHSS) in patients have polycystic ovarian syndrome (PCOS) and to prepare emergency fertility preservation in patients undergoing anticancer treatment, several researchers have reported IVM of oocytes retrieved from ovaries exposed by only hCG priming. However, the maturation rate and the developmental potential of embryos from IVM oocytes are significantly lower than those of oocytes matured in vivo. Here, we investigated the optimal time point for immature oocyte collection at post hCG only injection for in vitro maturation, in vitro fertilization and blastocyst formation. Immature GV oocytes were collected from 25 days old B6D2F1 female mouse at 12 hr, 14 hr, 16 hr or 24 hr post hCG injection. Oocytes were collected from antral or late secondary follicle by puncturing with 26 G needle. Collected oocytes were cultured in G2 medium with 10% FBS, FSH, estradiol, and hCG for 16 hr in vitro and subjected in vitro fertilization and further embryonic development. To examine follicular maturation, we estimated the numbers of primordial, primary, secondary follicle and antral follicle on ovaries of each time point post hCG. To confirm the optimal time point post hCG injection for collecting immature oocytes, we recovered the oocytes from each time point. There is no difference in the number of oocytes per mice. Oocytes collected at 14 hr post hCG injection were shown higher maturation rate to MII stage and blastocyst formation compare to other three groups (p<0.01). However, there is no difference in the maturation rate on the other three groups. Also, apoptotic signal with TUNEL assay or anti-PARP staining was not change in ovaries from all experimental groups. Granulosa cell proliferation test with anti Ki-67 or anti AMH was not show any difference. According to these results, there are no significant differences in four different time points at 12 hr, 14 hr, 16 hr or 24 hr of collection of immature oocytes in hCG primed mouse. However, oocytes from 14 hr post hCG injection showed higher percentages of maturation rate, in vitro fertilization rate, blastocyst formation.
Fifty-four Korean native and 28 foreign rice varieties harvested in 1998 and 1999 were examined for antioxidative activity that is measured to a chemiluminescence and superoxide radical intensity, by the flow injection chemiluminescence (FI-CL) system and an electron spin resonance (ESR) spectrophotometer, respectively. In the chemiluminescence measurement by FI-CL, radical scavenger activity did not differ significantly among rice varieties between origin types of rice varieties, and between storage periods. GinSun and Hongchoengdo, colored rice exhibited high electron scavenging effect by ESR. Therefore, these results indicate that the pigments of rice varieties may play important antioxidative roles and that it may be possible to breed rice varieties with higher antioxidative potentials.
Fifty-four Korean native and 28 foreign varieties harvested in 1998 and 1999 were examined for superoxide dismutase (SOD) and peroxidase (POD) activities. The SOD and POD activities of leaves extracts in Korean native and foreign rice varieties showed variation at the heading stage. The activities of SOD and POD changed with growth stage. In comparison of storage duration, the SOD and POD activities of the extract from three months stored seeds in Korean native (CV=53.3%) and foreign rice (CV=57.9%) varieties were higher than that of stored rices for a year in seed extracts although the activities among varieties did not show significant variation. Also, the averaged activity of foreign rice varieties (SOD=12.9%) was relatively higher than that of korean native rice varieties (SOD=10.7%). The test of activity at the enzymatic level related to antioxidative activity suggests that the rice varieties with higher antioxidative potentials can be developed and also may provide information with rice breeder to breed rice variety with a high antioxidative activity
This study was to investigate antioxidative activity of rice grain using 1, 1-Diphenyl-2-picrylhydrazyl(DPPH) and thiobarbituric acid (TBA) method and germination ability for screening rice varieties with high antioxidative activities on korean native and foreign rice varieties harvested in 1998 and 1999. The average antioxidative activity of foreign rice varieties (DPPH 63.5% and TBA 55.2%) was significantly higher than that of native rice varieties (DPPH 47.2% and TBA 45.6%) on varieties harvested in 1999. The promptness index (PI) of native rice varieties was higher in stored rices for three months (mean PI=160.7) than that of stored rices for a year (mean PI=141.6). On the other hand, the PI of foreign rice varieties was higher in stored rices stored for a year (mean PI=176.7) than that of stored rices for three months (mean PI=157.5). Varieties with high redness of hulled rice (a-value) showed significant lipid peroxidation inhibitory activity to DPPH in a stored rices for a year (r=0.5744** ) and stored rices for three months (r=0.5630** ) . These results indicate that the pigments of hulled rice varieties may play important antioxidative roles and colored rice varieties with higher antioxidative potentials can be developed and also may provide information with rice breeder to breed rice variety with a high antioxidative activity for a rapid screening of a small amounts of a large number of samples using color value.