최근 지구온난화에 의한 이상기상으로 겨울철 한파와 봄철 저온에 의한 농작물 피해가 심각하게 발생하고 있다. 특히 과수의 개화기 저온피해는 꽃눈의 생육단계에 따라 차이가 있으며 발육이 진전될수록 내한성이 약해져 개화 직전부터 낙화 후 1주까지 한계온도가 다르게 발생한다. 따라서 개화기가 빠른 사과 ‘홍로’가 ‘후지’보다 피해가 심각한 것이 일반적이나 2020년 4월 저온피해는 개화기가 늦은 ‘후지’의 피해가 심하게 발생하여 그 원인을 분석하고자 본 연구를 수행하였다. 충북 보은군 사과나무 ‘후지’와 ‘홍로’를 동시에 재배하는 2농가를 대상으로 품종 간 피해율 실태조사를 실시하였다. 또한 정확한 품종 간 비교 분석을 위하여 생육단계가 동일한 시료를 선택하여 인위적으로 저온처리(-2.0℃, -4.0℃)를 하여 피해 정도를 조사하고, 원인 분석을 위해 조직 내 유리당과 호르몬 함량을 분석하였다. 실태조사 결과 2농가 모두 ‘후지’가 ‘홍로’보다 피해율이 높았으며, 특히 B농가(저지대, 평지) ‘후지’의 경우 피해율이 60.5%로 가장 높았다. 또한 동일한 생육 단계의 시료를 사용한 인위적 저온 처리 시험결과에서도 ‘후 지’와 ‘홍로’ 품종 간 피해율에 유의한 차이가 있었다. 유리당 함량은 저온 피해율이 낮았던 ‘홍로’가‘후지’보다 높았으며, 호르몬 분석 결과 정상 꽃눈보다 손상된 조직에서 ABA, IAA 와 SA 함량이 높게 나타났다. 따라서 본 연구 결과 조직내 유리당 함량이 높으면 저온 피해율이 낮은 것을 확인할 수 있었으며, 특히 저온 피해율은 sorbitol 함량과 부의 상관관계를 이루고 있다.
The purpose of this study was to examine the effect of growing stages of the Korean Native Striped Bull (KNSB) on the freezability and fertility of frozen-thawed semen. First, we investigated the total motility (TM) and progressive motility (PM) according to the diluent used for semen freezing. Second, we examined the effect of the age of KNSB on semen volume, TM and PM of fresh and frozen-thawed semen. Third, we examined the effect of frozen semen from the different age of KNSB on the fertilization rate, and the artificial insemination pregnancy rate. The diluents used in this experiment were Triladyl and Tris-egg yolk extender (EYE). Semen was collected from 5 KNSB in the growing stage (15 months) and 5 adult KNSB (36 months). When Triladyl or Tris-EYE extender was used for semen freezing, there was no difference of the mean TM and the mean PM. However, the mean TM was significantly higher in Bull No. 1885 than Bull No. 4283 ( <0.05). The mean volume of semen collected from the 15-month-old bulls (2.3 ml) was significantly lower ( <0.05) than that from the 36-month-old bulls (5.0 ml). The mean semen concentration was similar for the 15-month-old ( spermatozoa/ml) and 36-month-old ( spermatozoa/ml) bulls. For the 15-month-old and 36-month-old bulls, the mean TM of fresh semen were 93.7% and 88.3%, respectively, and the mean PM were 97.0% and 88.3%, respectively; the 15-month-old bulls showed a particularly high PM ( <0.05). For the 15-month-old and 36-month-old bulls, the mean TM (56.0% and 58.0%, respectively) and the mean PM (64.0% and 70.7%, respectively) of frozen-thawed semen did not differ. The development rates of embryos after fertilization and the pregnancy rate after artificial insemination using frozen-thawed semen did not differ according to the bull's age. In summary, semen volume differed according to the bull's age, but semen concentration and survival rate, the fertilization rate, and the pregnancy rate did not differ according to the stripe bull's age. Accordingly, semen from bulls in the growing stage can be collected and frozen for the preservation and multiplication of rare livestock.
This studies were conducted to investigate the survival rate of frozen-thawed spermatozoa of Jindo Dog by monosaccharide and freezing rates. Experimental animals were prepared 12 males within 1~8 year's old and collected once in a couple of weeks by digital manuplation methods. Collected semen was diluted 1:1 with Tris-egg yolk extender and added 4, 6 or 8% of glycerol and none, 4 mM glucose or 4 mM fructose as cryoprotectant and was equilibrated for 2 hrs in . In monosaccharide groups, the freezing rate was 5 cm-5 min. above . The survival rates without monosaccharide were , , in 4, 6 or 8% glycerol, respectively. In addition of glucose, the survival rates were , , in 4, 6 or 8% glycerol, respectively and in fructose, were , , in 4, 6 or 8% glycerol, respectively. There showed significantly different between glycerol groups and monosaccharides groups (p<0.05). The survival rates of freezing rate in 5 cm-5 min. group was , , and in 10 cm-10 min. group was , , in 4, 6 or 8% glycerol, respectively. There were significantly different between freezing rates (p<0.05). These results suggest that the addition of fructose with 6%-glycerol and slow freezing improve the survival of frozen-thawed sperm in Jindo Dog.
본 실험은 돼지정액의 동결보존을 위한 희석액과 냉각속도 및 동해방지제의 적정농도를 결정하기 위해 실시하였으며, 얻어진 결과는 다음과 같다. 1. 에서 까지의 냉각속도에서는 LEY 희석액에서 분당 로 냉각하는 것이 생존율과 정상 첨체율에서 가장 높은 결과를 얻었다. 2. LEY 희석액이 BF5와 M-Soejima 희석액보다 정자를 동해로부터 보호하는 능력이 우수하였다. 3. LEY 희석액에 첨가하는 glycerol의 농도는 3 또는 가 의 glycerol
본 실험은 실험견의 정액 동결 시 희석 액에 첨가되는 Glycerol 농도, 동결속도, 동결보존액 첨가후 평형시간, 융해온도와 시간에 따라 정자의 생존성과 운동성을 조사하여 최적의 동결조건을 확립하기 위해 실시하였다. 1. 각기 다른 Glycerol 농도를 함유한 동결보존액에서 동결보존 후 융해하였을 때 4%의 Gly-cerol 농도에서 각각 68.87.4%, 73.28.3%로서 다른 군보다 유의하게 높은 생존율과 운동성 나타냈다(P<0.05). 2.
The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6 to -35 at -0.3 or -O.6 /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3 /min and -0.6 /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3 /min(63.6%) than in -0.6 /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3 /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.
This experiment was carried out to investigate the ovarian responses of the ovulation point, ovarian weight and size, the number of ovarian follicles and collected embryos, and to study the effects of the developmental stages (oocytes, 2-4 cell. 8-16 cell and morulae), additional levels of Ficoll (0, 15, 30%) on the survival rate (FDA-test) of rat embryos frozen in vitrification solution (20% glycerol + 10% ethylene glycol + 10% sucrose). Sunanarized results was as follows; 1. The mean ovulation point per head was 7, and the weight of ovaries was 0.03g. The size of ovary was 5.9 mm(L) and 4.6 mm(W), and the number of ovarian follicles over and below 2 mm was 4.7 and 8.7, respectively. The number of the collected embryos per head was 5.5 (79%). 2. 2. The FDA score of embryos frozen in 20 G 10 E 10 S without Ficoll was 2.8 (oocyte), 2.6 (2-4 cell), 3.9 (8-16 cell) and 3.6 (morula), respectively. However, there were no significant differences among treatments. 3. The FDA score of embryos frozen in 20 G 10 E 10 S with 15% Ficoll was 3.4 (oocyte), 4.0 (2-4 cell), 4.7 (8-16 cell) and 4.8 (morulae), respectively (P>0.05). 4. The FDA score of embryos frozen in 20 G 10 E 10 S with 30 % Ficoll was 3.7 (oocyte), 3.2 (2-4 cell), 4.4 (8-16 cell) and 4.4 (morulae), respectively (P>0.05). 5. As shown in the above results, the higher survival rate was obtained in the treatment of 15% Ficoll than that of 30%. And the survival rate (FDA-test)of the oocytes and 2-4 cell stages of the rat embryos was lower than that of 8~16 cell and morulae stages. It was considered that 8-16 cell and morulae could be available for the successful freezing by vitrification of rat embryos with 15% Ficoll except for oocytes.
This study were carried out to investigate the effective concentration of cryoprotective agents and sucrose by one-step straw method, and to determine the optimum thawing temperature and equilibration time of frozen porcine embryos. The porcine embryos foflowing dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3 water. Survival rate was defined by FDA test. The results are sunnnarized as follows : 1. The survival rates of porcine embryos after ultrarapid frozen4hawing in the freezing medium with a various concentration of glycerol, DMSO and propanediol added 0.25M sucrose were higher survival rate than those of sucrose concentration of 0.50M. 2. The survival rates of porcine embryos after ultrarapid ftozen4hawing in the freezing medium added 0.25M and 0.SOM sucrose were higher survival rate than those of sucrose concentration of 0.75M and 1.00M. 3. The temperature thawed at 2 and 3 resulted in a significantly higher embryos survival rate after 72 hrs in culture than did at 35. 4. The equilibration time on the survival rate of porcine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time(10~20 min.).