본 연구는 필리핀의 생물자원 및 농업에 이용되어지는 식물자원과 그 근연종의 다양성 및 관리 현황에 대한 최근 정보를 공유함으로써, 유용 유전자원의 확보와 유전자원 관련 국제기술 협력사업의 효율을 높이기 위한 기초자료를 제공코자 하며, 수행한 결과는 다음과 같다. 1. 필리핀은 7,107개의 크고 작은 섬들로 구성되어 있는 300,000 km2의 면적을 보유하고 있으며, 식물보유종으로 세계 5위를 차지할 정도로 다양성이 높은 국가이다. 2. 전 국토의 32% 정도의 면적이 경작지로 이용되고 있으며, 주요작물로는 벼, 옥수수, 코코넛, 사탕수수, 바나나, 마닐라삼, 망고 등이고, 벼멸구 내성을 지닌 O. offininalis 등 주요작물의 야생종 및 야생근연종들이 다수 분포하고 있다. 3. 필리핀 농업자원의 현지내 보존은 농가 보존형태이며, 야생종 보호를 위해 NIPAS 65개 구역을 지정하여 운영하고 있으나 작물연계한 생물다양성이나 생태시스템 관리 등 종합적인 프로그램은 부족하다. 4. 총 173,205점의 자원들이 현지외 보존되고 있으며, 이중형태특성평가는 40%, 생화학평가는 7%, 분자학적 평가는 3%, 병충해나 생산성 등 평가는 60%가량 수행되었다. 5. 식물자원의 보존과 관리를 위한 정부연구기관 및 네트워크가 설립되어 있으며, 식물유전자원에 관한 연구는 DA, DENR, 그리고 DOST 등의 국가적 조직들과 협력하여 수행하고 있다. 6. 공화국법령 8435 등 식물자원의 보존과 이용을 위한 상세한 내용의 필리핀 국내법이 마련되어 있다.

유전자원은 지역, 국가 및 국제적 수준에서 ex situ와 in situ의 상보적 보존을 통해 보존되어지고 있다. 유전자원은CBD에서 정의한 것처럼 유전적 정보와 지식을 포함한 물리적이고 실질적인 자원을 의미한다. 대만의 식물유전자원센터는 1993년 대만농업연구소 산하에 설립되었으며 2010년까지73,275점의 유전자원을 중·장기 보존소에 보존해왔다. 또한,포장유전자은행으로는 몇 개의 보존소에서 5,106점의 과수 및약용작물을 보존해왔으며 상보적으로 기내보존과 초저온동결보존을 이용하여 소실위험에 놓이거나 포장보존이 어려운 영양체 자원을 보존하고 있다. 또한 자원관리의 위험요인 및 안전관리, 종 동정 및 유전자원의 교배 및 상업적 이용 등에 대한 시행계획이 공식화되어 생물다양성에 기반한 생산물 시장개척에 기여하고 있다. 특히, 지역품종이나 육종계통 유전자원의 지속가능한 이용은 유전자원 관리의 규모에 따라 좌우된다고 할 수 있다. 유전자원의 효율적인 관리는 식량과 농업에서의 현재와 미래를 보장하는데 필수적이므로 유전자원을 보존 및 관리하는 전 세계적 노력에 참여함으로써 좀 더 큰 이익과책임을 기대할 수 있다. 특히, 지구온난화와 식량위기 등의 차원에서 효율적인 국내 및 국제적 식량 및 농업안보의 요소로서 유전자원 보존체계를 갖추고 관리하는데 적극적인 노력과실천이 필요하다.

Background : Ginseng seeds are one of short-lived seeds species which loose their viability easily in the condition of conventional storage. Cryopreservation using liquid nitrogen (LN) has been recommended as a alternative storage for this kind of germplasm short lived or dessiccation-sensitive. This study was performed to find out whether cryopreservation could assure initial viability not only for seeds with high germination rate but also for seeds with low germination rate (in aging process). Methods and Results : In this work, 3 cultivars of dehisced ginseng seeds were artificial aged in the condition of 40℃, 95% RH during 6 hours with 3 hours-interval. The germination rate of ginseng seeds was decreased to the range of 71 - 94% of initial viability by artificial aging treatment. After 24 hours of vapor-LN exposure on both of artificial aged and non treated seeds as a cryopreservation, germination rate for each cultivar was decreased with the range of 76 - 95% of initial viability. While the decreasing patterns of germination rate for each cultivar showed similar curves between before and after vapor-LN exposure, the aging effects could be slightly little by cryopreservation for 3 cultivars of ginseng seeds. Conclusion : From the above results, we may suggest that cryopreservation could be recommened for storage tool of dehisced ginseng seeds even with low viability also and expected to make slower seed aging process during preservation period through further study.

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.

Cryopreservation has become a powerful method of the assisted reproduction technology and supports fertility preservation of cancer and other indication patients. After controlled ovarian hyperstimulation, surplus oocytes and embryos were recommended to store using cryopreservation. Recently, vitrification is replaced with traditional slow freezing protocol, because of improved survival rates and clinical outcomes. Vitrification requires a high concentration of CPAs that may induce significant osmotic and metabolic damage to cells including oocytes even in a short exposure of a few minutes. Generally, MPF plays a crucial role in the cell cycle regulation and maintaining the meiotic arrest of oocytes. In fact, it has been observed to decline in MII ovine oocytes after vitrification and would be suggested that one of the main causes of low fertilization rate and developmental competence derived from cryoinjury during vitrification. Therefore, the aim of this study was to evaluate the effect of caffeine treatment on the activity of MPF, MAPK level in vitrified/warmed mouse mature eggs. Caffeine, Phosphataseinhibitor, may maintain active form of MPF. We evaluated their survival after warming procedure, fertilization, cleavage, and developmental rates. Ovulated MII eggs were retrieved from 6 weeks old B6D2F1 female mouse at 14hr post hCG injection. Collected MII eggs were maintained in HTF medium containing 10% KSR with or without caffeine for 1hr. Eggs were vitrified in 7.5%EG +7.5%DMSO equilibrium solution, 15%EG + 15%DMSO + 0.5M sucrose vitrification solution with or without caffeine. Also warming solution contained sucrose (0.5M, 0.25M, 0.125M, and 0M) with or without caffeine. After warming, eggs were cultured in HTF medium with or without caffeine for 2 hr then fertilized with epididymal sperm in vitro and cultured in KSOM for 5 days to analyze embryonic development. Survival rates were similar in all experimental groups. However, fertilization rate was higher in with caffeine group compare to without caffeine significantly (80% vs. 85%, p<0.05). 2-cell and blastocyst formation were increased in caffeine group (p<0.05). MPF activity and MAP kinase activity were recovered in with caffeine group after vitrification/warming process. In conclusion, Caffeine may maintain MPF and MAPK level in vitrified/warmed MII eggs, and enhance fertilization and further embryonic development.

The genus Vicia comprises 166 annual or perennial species distributed mainly in Europe, Asia, and North America, also extending to the temperate regions of South America and tropical. However, utilization of SSR markers have not been investigated extensively in Vicia species as compared to other crop species. Here, we have assessed the potential for transferability (cross-species amplification) of cDNA microsatellites markers developed from common vetch (Vicia sativa subsp. sativa). In total, 226 alleles were detected in 36 microsatellite loci. The number of alleles per marker ranged from one to 20, with an average of 6.3. The gene diversity and polymorphism information content value averaged 0.540 and 0.503, with a range of 0–0.85 and 0–0.84 respectively. For transferability of the SSRs, amplification was carried out with selected from two to 8 accessions of 22 different Vicia species. For individual species, the successful amplification rate ranged from 32.6% in V. ervilia to 81.9% in V. sativa subsp. nigra, with average of 48.8%. As the rate of successful amplification of microsatellite markers generally correlates with genetic distance, these SSR markers are potentially useful in the analysis of genetic relationships between or within Vicia species.

The objective of this study was to rapidly evaluate fatty acids in a collection of foxtail millet (Setaria italica (L.) P. Beauv) of different origins so that this information could be disseminated to breeders to advance germplasm use and breeding. To develop the calibration equations for rapid and nondestructive evaluation of fatty acid content, near-infrared reflectance spectroscopy (NIRs) spectra (1104-2494 nm) of samples ground into flour (n=100) were obtained using a dispersive spectrometer. A modified partial least-squares model was developed to predict each component. For foxtail millet germplasm, our models returned coefficients of determination (R2) of 0.91, 0.89, 0.98 and 0.98 for strearic acid, oleic acid, linoleic acid, and total fatty acids, respectively. The prediction of the external validation set (n=10) showed significant correlation between references values and NIRs values (r2=0.97, 0.91, 0.99 for oleic, linoleic, and total fatty acids, respectively). Standard deviation/standard error of cross-validation (SD/SECV) values were greater than 3 (3.11, 5.45, and 7.50 for oleic, linoleic, and total fatty acids, respectively). These results indicate that these NIRs equations are functional for the mass screening and rapid quantification of the oleic, linolenic, and total fatty acids characterizing foxtail millet germplasm. Among the samples, IT153491 showed an especially high content of fatty acids (84.06 mg g-1), whereas IT188096 had a very low content (29.92 mg g-1).