The present study was conducted to examine the effect of soybean silage as a crude protein supplement for corn silage in the diet of Hanwoo steers. The first experiment was conducted to evaluate the effect of replacing corn silage with soybean silage at different levels on rumen fermentation characteristics in vitro. Commercially-purchased corn silage was replaced with 0, 4, 8, or 12% of soybean silage. Half gram of the substrate was added to 50 mL of buffer and rumen fluid from Hanwoo cows, and then incubated at 39°C for 0, 3, 6, 12, 24, and 48 h. At 24 h, the pH of the control (corn silage only) was lower (p<0.05) than that of soybeansupplemented silages, and the pH numerically increased along with increasing proportions of soybean silage. Other rumen parameters, including gas production, ammonia nitrogen, and total volatile fatty acids, were variable. However, they tended to increase with increasing proportions of soybean silage. In the second experiment, 60 Hanwoo steers were allocated to one of three dietary treatments, namely, CON (concentrate with Italian ryegrass), CS (concentrate with corn silage), CS4% (concentrate with corn silage and 4% of soybean silage). Animals were offered experimental diets for 110 days during the growing period and then finished with typified beef diets that were commercially available to evaluate the effect of soybean silage on animal performance and meat quality. With the soybean silage, the weight gain and feed efficiency of the animal were more significant than those of the other treatments during the growing period (p<0.05). However, the dietary treatments had little effect on meat quality except for meat color. In conclusion, corn silage mixed with soybean silage even at a lower level provided a greater ruminal environment and animal performances, particularly with increased carcass weight and feed efficiency during growing period.
Bulbophyllum auricomum Lindl. is a rare orchid and has flowers with an attractive fragrance. The present study investigated the tissue culture method for micropropagation. Capsules derived from artificial self-pollination were obtained for the best seed germination in MS basal medium. Plant growth regulators (1.0 mg·L-1 of BAP and 2.0 mg·L-1 of NAA) were affected by callus induction from subcultured pseudobulb explants. For the callus subculture, different natural plant extracts were tested in 11 treatment media. Among them, MS medium with 150 mL·L-1 of coconut water was generally effective in fresh weight (1.75 ± 0.08) and (3.01 ± 0.20) of callus proliferation and PLBs induction at 1 and 2 months, respectively, followed by an MS combination of 30 g·L-1 of banana and 20 g·L-1 of potato extract. The results of a comparative study of different MS mediums containing plant growth regulators with a natural extract combination and MS medium supplemented with natural extract only showed that MS medium supplemented with a combination of natural extracts (150 mL·L-1 of coconut water) and plant growth regulators (2.0 mg·L-1 of BAP and 1.0 mg·L-1 of NAA) obtained the highest shoot regeneration (3.37 ± 0.17) and (6.41 ± 0.68) after 1 month and 2 months of culturing, respectively.
이 연구의 목적은 근육 세포의 증식 배양을 위해 필요한 핵심 인자인 혈청이 혈청 대체물의 첨가에 의해 대체될 수 있는지를 분자 생물학적 측면에서 검증하는 것이다. 혈청 대체물이 첨가된 low serum (5% FBS) 기반의 promo cell 배지는 성장 배양액으로 사용되었고, 마우스 하지 골격근의 근육세포는 pre-plating (pp) 방법에 의해 분리되었다. Pre-plating 4의 세포는 작고 둥근 형태의 굴절성을 가진 Myoblast/satellite like cells의 형태가 관찰되었으며, 근육 줄기세포 전사인자들(Pax3/7, Myf5, Myod1)과 골격근 발달 전사인자(Myog)의 발현량이 섬유아세포와 비교하여 높게 나타났다. 따라서 그들을 Myoblast derived cells로 명명하고, 기본 세포주로 사용하였다. 분리된 MDCs는 5%, 10% 혹은 20% 혈청이 첨가된 배양액에서 2주간 배양되었다. 배양 6일째부터 대조군(5% 혈청)과 비교하여 20% 혈청은 세포 수가 증가하였으며, 양적 혈청 농도 의존성이 확인되었다. 증식 및 세포사멸 관련 유전자들은 대조군과 비교하였다. 배양 1주 차에 20% FBS군은 세포증식 촉진 유전자인 Myc 발현이 증가한 반면 pro-apoptosis 유전자인 Bax의 발현량이 감소했고, 2주 차에는 세포주기정지 인자인Cdkn1a의 감소와 Myc의 지속적인 증가와 Bax/Bcl의 감소가 나타났다. 각기 다른 FBS 처리농도에서 배양된 Myoblast derived cells을 동일한 Myotube유도 배양액에서 2주간 분화하였다. 대조군과 비교하여 20% FBS군은 Myod1, Myog, Myf6, Myh1의 유의적 증가가 1주부터 확인되었다. 결론적으로 혈청 대체 물질은 근육세포 배양액에서 혈청의 효과를 완벽히 모사할 수 없음이 증명되었고, 따라서 체외에서 상업적 목적의 근육 세포 대량 증식 및 분화를 위해서는 적절한 혈청 대체물 개발이 선행돼야 할 것으로 사료된다.
고구마 바이러스 무병묘의 기내급속증식을 위한 적정 생장 조절물질 및 sucrose 농도, 최소생장 기내보존(15°C)에 미치는 광의 영향과 생존율 및 기내생장 특성 등을 조사하였다. 고구마 무병묘의 마디배양은 0.2mg·L-1 BA 첨가배지에서 줄기신장, 줄기직경, 잎수, 뿌리수, 생체중 및 건물중 등이 가장 양호하였다. 배양부위 및 배지물리성에 따른 적정 sucrose 농도는 마디배양은 5% sucrose를 첨가한 고체배지에서, 정단배양은 3% sucrose를 첨가한 액체배지에서 줄기두께, 잎수, 뿌리수, 뿌리길이, 생체중 및 건물중 등의 생육에 가장 효과적이었다. 15°C 저온항온기에서 고구마 무병묘의 최소생장 기내보존은 암상태에서는 3개월 내에 모두 고사하였으나, 적색:청색(7:3) 혼합 LED (150±5μmol·m-2·s-1 PPFD)에서는 5개월까지 100% 생존하였다. 따라서 고구마 무병묘의 최소생장 기내보존에는 광이 필요하며, 샬레에 밀식(10 개체/샬레)할 경우, 좁은 공간에서 대량보존이 가능하였다.
차나무의 기내배양 과정 중 증식배양 단계에서 LED 광질 조건에 따른 기내배양묘의 생육 특성과 광합성에 미치는 영향을 조사하였다. 광질은 적색광(R), 청색광(B), 혼합광(R+B+W)을 사용하여 처리하였고, 형광등(F)을 대조구로 하였다. 초장 생육은 적색광에서 가장 좋았으며, 특히 뿌리 생육에 있어서 혼합광은 길이와 표면적 증대에 효과적인 것으로 나타났다. 또한 T/R율, 엽록소 함량은 혼합광 처리에서 증가하는 것으로 나타났다. 엽록소 형광반응 이미지 촬영 결과 모든 처리구에서 광질에 따른 Fv/Fm의 값은 현저한 차이가 없었다. 그러나 배양묘의 NPQ는 청색광 처리에서 가장 많이 증가하여, 다른 광질과 유의적인 차이를 보였으며, 광합성 효율을 억제시키는 것으로 나타났다. 이상의 결과로 차나무 기내 배양은 배양목적에 따라 광질을 선택하는 것은 매우 중요하며, 차나무 기내배양 시 건실한 식물 생산을 위해서는 혼합광 처리가 유리할 것으로 판단된다. 본 연구결과는 차나무 대량증식 및 우량묘 생산 등에 기초자료로 활용할 수 있을 것으로 판단된다.
The objective of this study was to establish an in vitro culture system for ovarian preantral follicles of B6D2F1. First, we optimized the in vitro preantral-follicle culture by culture duration, follicle stimulating hormone (FSH) type, and activin A concentration. Duration of in vitro culture for 9, 11, and 13 days was sufficient for the normal development of preantral follicles to antral follicles. Formation of cumulus cell–oocyte complex (COC) was induced by treatment with human chorionic gonadotropin (hCG; 2.5 IU/mL) and epidermal growth factor (EGF; 5 ng/mL). In addition, metaphase II (MII) oocytes formed during this in vitro culture of preantral follicles. In vitro preantral-follicle culture for 9 days showed higher rates of growth and maturation, thus yielding a greater number of antral follicles, and there were significant differences (p < 0.05) in the number of MII oocytes (that formed from these preantral follicles via differentiation) between the 9-day culture and 11-day or 13-day culture. The follicles cultured for 9 days contained a tightly packed well-defined COC, whereas in follicles cultured for 11 days, the COC was not well defined (spreading was observed in the culture dish); the follicles cultured for 13 days disintegrated and released the oocyte. Second, we compared the growth of the preantral follicles in vitro in the presence of various FSH types. There were no significant differences in the growth and maturation rates and in differentiation into MII oocytes during in vitro culture between preantral follicles supplemented with FSH from Merck and those supplemented with FSH from Sigma. To increase the efficiency of MII oocyte formation, the preantral follicles were cultured at different activin A concentrations (0 to 200 ng/mL). The control follicles, which were not treated with activin A, showed the highest rate of differentiation into antral follicles and into MII oocytes among all the groups (0 to 200 ng/mL). Therefore, activin A (50 to 200 ng/mL) had a negative effect on oocyte maturation. Thus, in this study, we propose an in vitro system of preantral-follicle culture that can serve as a therapeutic strategy for fertility preservation of human oocytes for assisted reproductive medicine, for conservation of endangered species, and for creation of superior breeds.
This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.
In this study, the principal objective was to investigate the effect of light quality and vessel ventilation on the growth and development, physiology, activities of antioxidant enzymes, and contents of mineral nutrients of carnation (Dianthus caryophyllus L.) ‘Marble Beauty’. Single node cuttings stuck into the plant growth regulator (PGR)-free MS medium in containers covered with caps with or without a ventilation filter were cultured first four weeks under white and then additional four weeks under either white (control), blue, red, or red + blue light emitting diodes (LEDs) for 56 days. Interestingly, a ventilated culture condition not only reduced the percentage of the hyperhydricity, but also increased the total chlorophyll content (Chl a + Chl b) of the plantlets as compared to the non-ventilated condition. In addition, blue LEDs produced plantlets with the greatest number of shoots and red LEDs produced plantlets with the greatest shoot length. The quality of plantlets was improved under a ventilation condition. Besides, under a ventilated condition, red + blue LEDs raised APX activity, and blue LEDs not only raised the activity of the CAT, but also increased tissue contents of such elements as K, Ca, Mg, Zn, Mn and Fe. The red LEDs increased contents of B and Si under a ventilated condition, and Na accumulation under a non-ventilated condition. Thus, including blue or red LEDs as the light source in a ventilated culture condition will produce plantlets of carnation ‘Marble Beauty’ in vitro with improved quality.
This study was conducted to evaluate the effects of insulin and epidermal growth factor (EGF) in a in vitro growth (IVG) medium on oocyte growth, in vitro maturation (IVM) and embryonic development of pig oocytes derived from small antral follicles (SAF) less than 3 mm in diameter. SAF oocytes were cultured for 2 days to induce IVG in alpha-minimal essential medium supplemented with 1 mM dbcAMP and 15% (v/v) fetal bovine serum. After IVG culture, oocyte maturation was induced by culturing IVG oocytes in IVM medium for 44 h. IVM oocytes that extruded the first polar body were selected and induced for parthenogenesis (PA) by applying electric stimulus. SAF oocytes cultured under the insulin treatment showed a significantly increased (P < 0.05) nuclear maturation (73.8%) compared to those cultured with insulin and EGF (59.8%). After PA, the proportions of blastocysts based on the number of metaphase II oocytes were significantly higher (P < 0.05) in oocytes that were cultured for IVG with insulin, EGF, and insulin + EGF (32.4%, 35.2%, and 34.8%, respectively) than in control (22.9%). IVG oocytes treated with insulin showed an increased oocyte diameter (116.3 μm) compared to those treated with insulin and EGF (114.0 μm) (P < 0.05). Intra-oocyte GSH content significantly increased (1.07 pixels/oocyte) by insulin treatment during IVG compared to that of oocytes treated with insulin + EGF (0.78 pixels/oocyte). These results demonstrate that IVG culture of SAF oocytes under insulin or/and EGF treatment supports oocyte maturation and improves embryonic development to the blastocyst stage after PA in pigs.
The beneficial effect of silicon (Si) in increasing salt stress tolerance has been observed in many plants, including the cereal crops rice, wheat, and barley. In this experiment, we examined the effect of Si on the survival and growth of torenia (Torenia fournieri L inden ex F oum) ‘ Duchess Blue and White’ cultured in vitro in the presence and absence of salt stress. Previous reports had suggested that torenia exhibited low salt tolerance. Shoot buds isolated from 16-day-old seedlings were cultured on Murashige and Skoog (MS) medium containing 0, 50, or 100 mM NaCl alone or in combination with 1.8 or 3.6 mM Si supplied as K2SiO3. Plant survival rate was significantly reduced by NaCl supplementation compared with the control. The survival rate significantly increased to 100% when 1.8 or 3.6 mM Si was added to the MS medium containing 50 mM NaCl. However, only 31% of plantlets survived when 1.8 mM Si was added to the culture medium containing 100 mM NaCl. Shoot and root lengths significantly decreased with increasing NaCl concentration in the culture medium, whereas addition of NaCl to the MS medium also significantly reduced fresh and dry weights. However, Si supplementation significantly increased fresh and dry weights under 50 mM NaCl, compared with the control. The greatest fresh and dry weights were recorded when shoot buds were cultured on MS medium containing 50 mM NaCl and 3.6 mM Si. The activities of the antioxidant-scavenging enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT), but not peroxidase (POD), were markedly higher in the presence of 50 mM NaCl than the activity of the control. When Si was added to the medium containing 50 mM NaCl, activities of SOD, POD, APX, and CAT decreased as compared with the 50 mM NaCl treatment. Thus, Si-mediated tolerance to NaCl stress was not due to increased activity of antioxidant enzymes. Although Si was not effective in increasing tolerance to high salt concentrations, such as 100 mM NaCl, the results suggested that Si supplementation could effectively enhance tolerance to 50 mM NaCl stress.
내건성이 우수하다고 판명되어진 비수리(Lespedeza cuneata), 새(Arundinella hirta), 소리쟁이 (Rumex crispus), 장구채(Silene firma), 매듭풀(Kummerowia striata) 5수종의 기내대량증식 조건을 구명하였다. 5종의 선발종 중 비수리는 WPM배지에서 길이생장이 가장 좋았으며, 새는 MS배지, 소리쟁 이는 LP배지에서 장구채는 LP배지, 매듭풀은 SH배지에서 좋은 길이생장을 보였다. 뿌리발생도 수종에 따라 배양배지에 따라 다르게 나타났다. 비수리의 경우 기내발근은 모든 배지에서 뿌리가 발생하지 않 았으며, 새는 MS배지, 소리쟁이는 WPM배지, 장구채는 MS배지, 매듭풀은 1/2MS배지에서 가장 좋은 뿌리생장을 보였다. 잎 갈변은 비수리는 B5배지, 새는 WPM배지, 소리쟁이는 B5배지, 매듭풀은 LP배 지에서 가장 심한 갈변을 보였다. 생존율은 비수리, 소리쟁이, 장구채는 모든배지에서 100% 생존율을 보였으며, 새는 WPM배지에서 25% 생존율을, 매듭풀은 LP배지에서 25%의 낮은 생존율을 나타냈다. 선 발종의 다경줄기를 유도하기 위해 cytokinin을 처리한 결과 비수리의 경우 TDZ 0.5mg/L 처리구, 새는 BA 0.5mg/L, 소리쟁이는 TDZ 0.1mg/L, 장구채는 BA 0.5mg/L, 매듭풀은 BA 0.5mg/L 처리구에서 가장 많은 다경줄기를 형성하였다. cytokinin 처리는 줄기생장, 기내발근 및 다경줄기 유도에 영향을 미쳤다. 기내에서 배양된 식물체는 4종의 인공상토에서 성공적으로 순화되었다. 본 연구 결과들은 내건 성을 가진 5종의 유용식물의 품종개발 및 대량증식에 크게 기여할 수 있을 것으로 판단된다.
반하(Pinellia ternata(Thunb.) Breit)는 천남성과에 속하는 다년생 초본식물로서 조직배양을 이용하 여 대량번식 방법 연구가 활발히 진행되어 왔다. 하지만 대량생산된 괴경들의 토양 순화 및 적응을 위 한 환경 조건의 확립의 연구는 미비하다. 따라서 본 연구에서는 기내에서 증식된 반하의 괴경을 이용하 여 우수한 품질의 약재와 건전묘의 대량생산을 위해 생육에 적합한 토양조건을 탐색하였다. 본 연구에 서는 액체배지에서 현탁배양 한 괴경(Type 1)과 고체배지에 치상하여 배양된 괴경(Type 2)두 종류를 비 교하였다. 토양은 3개의 조성으로 조합하여 생육을 비교하였으며, 코코피트, 피트모스, 버미큘라이트, 펄라이트 및 제오라이트를 배합하여 사용하였다. 기내 배양 조건이 다른 처리구들의 토양 조성별 순화 율 측정하였으며, 생육의 차이를 확인하기 위해 8주동안 생육 후 초장, 잎 수, 마른잎 수, 괴경 수, 괴 경 크기, 생체중 및 건물중을 측정하였다. 또한 주아의 생성율을 확인하기 위하여 4주와 8주에 측정을 진행하였다. 그 결과 Type 2가 펄라이트를 20%증량한 상토 B에서 가장 우수한 생육을 보였다. 괴경의 비대에 영향을 주고 묘로서 이용을 위하여 필요한 잎의 수는 상토 B에서 가장 많은 1.7개로 나타났고 가장 잎의 출현이 없었던 Type 1의 상토 C가 0.9개로 나타나 1.8배의 차이를 나타냈다. 또한 같은 처 리구에서 건물중이 통계적으로 유의한 차이를 보이며 우수한 것으로 나타났다. 반하의 주아 생성율의 차이는 상토 B에서 1.1개로 우수하였으며, 가장 저조한 처리구의 수치보다 1.2배 높은 생성율을 확인할 수 있었다. 이러한 결과로 기내에서 배양된 반하의 토양 순화 및 적응기에 괴경의 비대를 유도하여 우 수한 한약재 생산이 가능 할 것으로 생각되며 차후 토양에서 재배 시 반하의 대량번식에도 도움이 될 것이라 사료된다.
Oocytes from small antral follicles (< 3 mm in diameter; SAFO) show lower developmental competence compared to those from medium antral follicles (3-8 mm in diameter; MAFO) in pigs. This study was designed to evaluate the effect of various macromolecules such as fetal bovine serum (FBS), porcine follicular fluid (PFF), bovine serum albumin (BSA) and polyvinyl alcohol (PVA) in in vitro growth (IVG) medium on oocyte growth, maturation, and embryonic development after parthenogenesis (PA). The base medium for IVG was α-MEM supplemented with dibutyryl cyclic AMP, pyruvate, kanamycin, hormone. This medium was further supplemented with 10% FBS, 10% PFF, 0.4% BSA, or 0.1% PVA. The in vitro maturation (IVM) medium was medium-199 supplemented with 10% PFF, cysteine, pyruvate, epidermal growth factor, kanamycin, insulin, and hormones. SAFO were cultured for 2 days for IVG and then cultured for 44 h for IVM. After IVG, the mean diameter of SAFO treated with FBS, PVA, and no IVG-MAFO (114.1, 113.0, and 114.8 μm, respectively) was significantly larger (P<0.01) than that of no IVG-SAF (111.8 μm). Oocyte diameter after IVM was greater (P<0.01) in SAFO treated with FBS, BSA and PVA (112.8, 112.9 and 112.6 μm, respectively) than other groups (110.4, 109.6, and 109.8 μm for no IVG-MAFO, no IVG-SAFO and PFF, respectively). Intraoocyte GSH content was not influenced by the macromolecules in IVG medium (0.92, 0.93, 1.05, and 1.12 pixels/oocyte for FBS, PFF, BSA and PVA, respectively). The proportion of oocytes reached the metaphase II stage was higher in PFF (73.6%) than in BSA (43.5%) and PVA (53.7%) but not different from that of FBS treatment (61.5%). The cumulus expansion score of oocytes after IVG was significantly influenced (P<0.01) by the macromolecules (2.94, 2.24, 1.84, and 1.38 for PFF, FBS, PVA, and BSA treatments, respectively). Blastocyst formation of PA oocytes that were treated with FBS (51.8%), PFF (50.4%), and PVA (45.2%) during IVG was higher (P<0.05) than that of BSA-treated oocytes (20.6%) but was not significantly different from that (54.8%) of no IVG-MAFO oocytes. Our results demonstrated that growth, maturation, and embryonic development of SAFO are greatly influenced by macromolecules in IVG medium and that PFF or FBS can be replaced with a chemically defined synthetic macromolecule PVA.
Somatic cell nuclear transfer (SCNT) technique is a key point of producing transgenic animal disease models. During in vitro production of SCNT embryo, the quality of matured oocytes are one of the important factors that regulate embryo developmental capacity. In preliminary test, we confirmed the effect of fibroblast growth factor 10 (FGF10) on porcine oocyte maturation. In this study, we investigated the developmental potential of SCNT embryos treated with the 10 ng/ml FGF10 (10 F) during in vitro maturation of recipient oocytes. The polar body emission rate was significantly higher in the 10 F treated group than control group. After SCNT, although the rate of fusion was no significant difference, the rate of cleavage and blastocyst formation was significantly increased in the 10 F treated group (p<0.05). In 10 F treated group, the total cell number was increased and the percentage of apoptotic cell was decreased in the blastocyst stage at day 7 (p<0.1). The transcription level of apoptosis relative gene, Casp3 was significantly decreased, while anti-apoptosis gene BCL2l1 was increased in the 10 F treated group compared to control group. The 10 F treated group was highly expressed the reprogramming related genes, Sox2 and POU5f1. Also, the first cleaving time was more faster and the percentage of cell block was significantly lower in 10 F treated group than in control group. In this study, we confirmed that 10 ng/ml FGF10 has effect on enhance the oocyte maturation and developmental capacity. These results demonstrate that FGF10 treatment can be used for in vitro development of porcine SCNT embryos and subsequent production of transgenic animal model.
Carbon source, an essential nutrient for plant growth, mainly includes exogenous sugar and CO2 of the environment in vitro. Therefore, the exogenous sugar and CO2 of the environment make the important roles in tissue culture. The aim of this study is to investigate the effects of different sugar concentrations (0, 10, 15 and 30 g·L-1) on the growth of colored Zantedeschia in vitro under certain CO2 concentration and explore the optimal sugar concentration. The plantlets in vitro of colored Zantedeschia had the largest root number, root weight, and root vigor under 0 g·L-1 (sugar-free culture) treatment. And they had the largest plant height, leaf length and leaf chlorophyll content, but p oor r oot v igor under 3 0 g·L-1 sugar. This study indicated that the optimal condition for proliferation and seedling culture of colored Zantedeschia plantlets in vitro was MS medium with 30 g·L-1 sugar, and the suitable medium for rooting culture and transplanting of colored Zantedeschia was MS medium with sugar-free culture under CO2 enrichment condition.
Sargassum fusiforme has traditionally been widely consumed in Asia as a food, and it has gained much attention due to its high nutritional, pharmaceutical, and industrial value. This study aimed to examine the promotional effects of ethanol extract (ET) and fraction obtained from ethyl acetate (FR) of S. fusiforme on hair growth in C57BL/6 mice and HaCaT cells. Five-week-old mice were used to compare hair regrowth during application of ET and FR for 21 days. Hair regrowth was evaluated by macroscopic observation and verified by hematoxylin-eosin tissue staining. Levels of mRNA expression of factors relevant to the hair growth cycle such as keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), and transforming growth factor-beta1 (TGF-β1) were examined by quantitative polymerase chain reaction (qPCR). Our results showed that ET and FR successfully promoted hair regrowth in shaved C57BL/6 mice at a dose >20 mg/kg. Moreover, ET and FR were effective in stimulating expression of KGF and VEGF mRNAs in a dose-dependent manner, whereas TGF-β1 was not activated. These results indicate that ET and FR of S. fusiforme effectively promoted hair growth and gene expression relevant to hair growth cycles in both in vitro and in vivo models.
본 시험은 효소제 및 효모추출물 급여가 사양성적, in vitro 반추위 발효 및 혈액성상에 미치는 영향을 구명하기 위해 23개월령의 한우 거세우 48두를 공시하여 사료용 첨가제로 농후사료의 0.1% 수준으로 급여하여 출하시까지 대조구(무첨가), 처리구 1(효소제), 처리구 2(효모추출물), 처리구 3(효소제+효모추출물)으로 구분하여 급여하였다. 사양성적에서는 급여 4개월과 8개월의 일당증체량과 사료요구율이 처리구 1에서 가장 향상되었고 (p<0.05), 대조구에서 가장 낮았다(p<0.05). 도축성적에서는 육량지수를 포함하여 처리구 1>처리구 3>처리구 2>대조구 순으로 나타났다. in vitro 배양액에 따른 처리구별 pH는 대조구>처리구 2>처리구 3>처리구 1 순으로 유의적(p<0.05)인 차이를 나타냈고, VFA와 사료소화율은 처리구 1과 3에서 높았고 (p<0.05), 대조구에서는 낮았다(p<0.05). 혈액성상에서는 면역 첨가제 급여군인 처리구 2와 3에서 급여기간에 따른 백혈구 수치가 유의적(p<0.05)으로 감소하였는데, 이는 면역글로블린 G의 유의적(p<0.05)인증가에 따라서 상대적으로 나타난 것으로 판단된다. 결과적으로, 한우 비육후기 거세우의 사료첨가제 급여 시험에서 처리구 1은 급여 4개월부터 대조구 대비 사양성적 및 도체성적을 향상시켰고, 처리구 2는 급여 2개월부터 면역에 관련된 면역글로블린 G의 혈액성상을 개선시키는 것으로 분석되었다.
돌나물 유전자원의 기내 활성보존을 위하여, 10mm 크기의 기내배양 shoot를 agar, Gelrite, ABA 및 sucrose 농도를 달리한 MS 배지에 치상하여, 4oC와 25oC에서 계대배양 없이 보존하였다. 배지는 0.2mg · L−1 BA를 기본으로 첨가하였고, agar와 Gelrite 배지에는 5% sucrose, ABA 배지에는 5% sucrose와 1.2% agar, sucrose 배지 에는 1.2% agar를 각각 첨가하였다. 상온 활성보존(25oC) 에서 sucrose와 Gelrite 배지는 생장억제 효과가 거의 없었고, 0.2mg · L−1 BA + 10mg · L−1 ABA + 1.2% agar, 또는 0.2mg · L−1 BA + 1.6% agar를 첨가한 배지가 효과적이었으며, 계대배양 없이 10개월까지 활성보존이 가능 하였다. 저온 활성보존(4oC)에서 12개월후 생존율은 모든 배지에서 100%였으며, 10mg · L−1 ABA, 또는 6% sucrose 첨가배지에서 계대배양 없이 18개월 이상의 활성보존이 가능하였다. 특히 고농도 sucrose 배지는 저온 활성보존(4oC)에서 돌나물 유전자원의 장기간 활성유지에 가장 효과적이었다.
거베라의 조직배양과정 중 증식배양단계에서 LEDs 파장이 기내배양묘의 생육에 미치는 영향을 조사하고 자 이 실험을 수행하였다. 절화용 거베라 ‘바네사’의 증식배양 시 발광다이오드(LEDs) 광질에 따른 생육특 성을 보면 주당 신초수가 red와 red(4) + blue(1) LEDs에서 대조구인 형광등에 비해 많았고 far-red에서 는 다소 증가 또는 blue LEDs에서는 줄어드는 경향이 었으나 그 차이는 통계적 유의성이 없었다. 초장은 red LEDs에서 가장 길었으며 주당 엽수는 red(4) + blue(1) LEDs에서 가장 많았고 엽면적은 far-red LEDs를 제외 하고는 비슷한 경향이었다. 주당 생체중과 건물중은 red LEDs에서 가장 무거웠으나 red(4) + blue(1) LEDs, 형광 등과도 큰 차이가 없었다. 증식배양단계에서 가장 중요 한 것은 신초를 확보하는 것으로 다른 생육특성들이 형 광등 대비 큰 차이가 없는 것으로 보아 red LEDs 또 는 red(4) + blue(1) 혼합 LEDs를 광원으로 사용하는 것이 적당하다고 판단된다.