• 생명과학회지
• /
• 제11권4호
• /
• pp.311-315
• /
• 2001

To identify the effects embryo developmental stage and gelrite concentration on plant regeneration in seed culture of rice, mature and immature seeds of rice were cultured on the $N_{6}$ medium supplemented with2 mg/$\ell$ 2.4-D and different levels of gelrite(0.2~1.0%). The calli formed immature embryos were produced more plants than those from mature embryos. The maximum frequency of plant regeneration was achieved in the culture of the calli of immature embryos which was harvested at the 21$^{th}$ day after pollination. The plant regeneration on the medium with gelrite was more accelerate than that on the medium with agar. The highest frequency(55%) of plant regeneration was obtained from the calli transferred to the medium with 6g/$\ell$ gelrite.

• Journal of Species Research
• /
• 제6권1호
• /
• pp.68-75
• /
• 2017

We investigated the effects of sodium hypochlorite (NaOCl) and culture medium on embryo swelling and germination of Calanthe discolor Lindl., and established a method for determining the swelling and protocorm formation of C. discolor seeds via in vitro examination of immature seeds. Treatment of immature seeds with NaOCl greatly enhanced the extent of embryo swelling and protocorm formation of immature zygote embryos compared to seeds without NaOCl treatment. The effects of the culture media were also evaluated with regard to embryo swelling and protocorm formation of in vitro cultured seeds with and without NaOCl treatment. Additionally, the effects of white fluorescent light and red and blue LEDs lights on seedling growth in in vitro culture were examined. The most suitable condition for seedling growth after 12 weeks of culture was the red LEDs light with POM medium. These results show effective asymbiotic germination and growth of C. discolor.

• 한국수정란이식학회지
• /
• 제13권2호
• /
• pp.147-157
• /
• 1998

This study was conducted to investigate the effects of in vitro fertilization, culture and embryo development according to in vitro maturation rate, protectant composition and equilibrium time after frozen /thawing of bovine immature oocytes. This results obtained in studies on the effect of different cryoprotectants on the viability, maturation and development of in vitro bovine oocytes were as follow: 1.The post-thawing of immature oocytes matured to metaphase II during culture time for 0 to 26 h, and those group (62~3%) were low than control group (76.7%). The optimal maturation time of frozen-thawed immature oocytes was at 24 h. 2.The viability of cryopreserved immature oocytes was not affected by sort of cryoprotectants. The developmental competence of frozen4hawed oocytes was not affected by cryoprotectants. These results indicate that an optimal maturation time of frozen /thawed immature oocytes was at 24h. Furthermore the viability of cryopreserved immature oocytes was not affected by sort of cryoprotectants and developmental competence of frozen /thawed oocytes.

• 한국작물학회지
• /
• 제40권3호
• /
• pp.308-313
• /
• 1995

본 시험은 미숙종자의 배배양을 이용한 보리의 세대촉진을 위하여, 재배조건(15/10, 25/15$^{\circ}C$)과 등숙기간에 따른 미숙종자의 형질을 조사하고 이들 형질들간의 상호 관계를 구명함과 동시에, 미숙종자의 배배양시 유묘의 생육 반응을 검토하였던 바, 그 결과를 요약하면 다음과 같다. 1. 출수 후 21일이 경과한 미숙종자의 배장은 이삭의 하부보다는 중상부가, 저온(15/10$^{\circ}C$)보다는 고온(25/15$^{\circ}C$) 조건에서 길었는데, 특히 저온 조건의 하부에서 짧았다. 그러나 부위별 차이보다는 생육 온도 조건에 따른 차이가 컸다. 2. 배배양 후 유아장과 근수 및 근장은 고온의 경우 이삭 부위별 차이가 적었으나, 저온조건에서는 중, 상부보다 하부의 생육이 현저히 불량하였다. 3. 등숙 기간에 따른 배장의 변리는 고온 조건에서 21일경에 최장에 이르러 더이상 신장하지 않았지만 저온 조건에서는 29일까지도 계속적으로 신장하여 저온 조건의 29일배가 고온 조건의 17일배와 유사한 경향치를 보였다. 4. 고온 조건에서 17일이 경과한 배의 배장은 입폭, 유아장, 근수 및 근장과 정의 상관이 인정되었으나, 21일이 경과한 것은 어느 형질과도 상관이 인정되지 않았다. 5. 미숙종자의 배배양을 이용한 세대촉진시에는 25/15$^{\circ}C$의 고온 조건에서 출수후 17일이 경과한 배를 이용하는 것이 적당할 것으로 보였다.

• 한국수정란이식학회지
• /
• 제24권3호
• /
• pp.145-150
• /
• 2009

This study was conducted to examine the effects of human follicular fluid and gonadotropin (FSH+HCG+rhEGF) on in vitro maturation, fertilization and development of human immature oocytes. Cumulus-oocyte complexes (COCs) were collected following for in vitro fertilization and embryo transfer (IVF-ET) cycles of the patients. At the time of oocytes collection, oocytes were classified into MII, MI and GV in accordance with their appearance (MII: Fully mature oocyte at metaphase II of meiosis; MI: Nearly mature oocytes at metaphase I of meiosis; GV: Immature oocytes at prophase I of meiosis). After controlled ovarian stimulation using gonadotropin(FSH) and human chorionic gonadotropin (HCG) in 70 ICSI cycles, 158 MI to MII matured oocytes were intracytoplasmic sperm injection (ICSI) ${\sim}4$ h after in vitro culture and 553 MII oocytes were ICSI after denudation. The aspirated MI and GV oocytes were cultured in culture medium containing 10% (v/v) serum protein substitute (SPS), 10% (v/v) human follicular fluid (hFF) and 10% (v/v) serum protein substitute (SPS)+1 IU/ml FSH+10 IU/ml HCG+10 ng/ml recombinant human epidermal growth factor (rhEGF). The maturation rate of immature oocytes was similar among the three group. When maturation medium was supplemented with 10% SPS, 10% hFF or gonadotropins, the fertilization rate of in vitro matured oocytes was higher in 10% SPS (80.0%), but there was no statistical significance (78.2%; hFF, 76.9%; gonadotropin, p>0.05). The development rate of human embryos developed to $6{\sim}8$ cells were not significant difference in the medium containing SPS, hFF and gonadotropins (65.6%, 65.9% and 66.7%). The results of these study suggest that human follicular fluid and gonadotropins supplemented in the culture medium was not effected on the in vitro maturation, fertilization and development of human immature oocytes.

• 식물조직배양학회지
• /
• 제22권4호
• /
• pp.207-211
• /
• 1995

고추냉이의 미숙배를 어뢰형 배(stage I )와 자엽배의 (stage II )로 구분하여 캘러스, 배발생 및 식물체 재분화에 미치는 생장조절제의 효과와 치상적기를 조사하였다. 캘러스 및 배발생은 미숙배의 자엽부분에서 관찰되었는데 캘러스의 양상에 따라 기관분화와 배발생이 이루어졌다. 캘러스 형성률은 stage I의 경우 1.0 mg/L IAA에서, stage II의 경우 1.0 mg/L 2,4-D와 0.1 mg/L BA 혼용처리구에서 가장 양호하였다. 체세포배 발생은 stage I의 미숙배는 생장조절제 무처리구에서, stage II의 미숙배는 1.0 mg/L IAA 처리구에서 가장 높았으며 캘러스를 통하거나 자엽에서 직접 발생되었다. Stage II의 미숙배가 stage I에 비해 캘러스 및 체세포배 발생이 왕성하였다. 이들 체세포배는 동일배지에서 계대배양하여 정상적인 식물체로 재생되었다.

• Plant Resources
• /
• 제7권2호
• /
• pp.87-92
• /
• 2004

Five different poplar hybrids were tested for rescuing embryo to elongate in vitro plantiets after hybridization. Ovaries and ovules were cultured on Woody Plant Medium (WPM) supplemented with cytokinins, 6-benzylamine (BA) and zeatin. Multiple shoots were initiated from half section of capsule with immature embryos after 21 days from pollination and tiny shoots were formed after the expansion of cotyledons in ovule cultures. Germinating response was better in intraspecific hybrids $(6.53\pm1.66)$ than interspecific crosses $(0.93\pm0.54)$ from half section of capsules on WPM medium. In general, zeatin was better than BA in inducing multiple shoots from isolated ovules. The highest average number $(19.40\pm4.53)$ of shoots was produced from immature ovules of 21 days post-pollination of WPM medium supplemented with 5.0 mg/L zeatin. The highest percentage of germination was 93% from the half section of in vitro ovary cultures. Soil acclimation was successfully conducted in cell tray containing artificially mixed soil with 96% survival rate.

• Asian-Australasian Journal of Animal Sciences
• /
• 제24권11호
• /
• pp.1541-1546
• /
• 2011

This study was carried out to examine the influence of minimum essential medium (MEM) vitamins supplementation to in vitro maturation medium and in vitro culture medium on the development of porcine embryos. Porcine embryo development was investigated following cultivation in both in vitro maturation and culture medium with the supplementation of MEM vitamins (0, 0.1, 0.2 and 0.4%) using immature oocytes collected from the ovary of prepubertal gilts. Embryo development was observed and the total cell number in each blastocyst generated under the culture conditions was quantified following supplementation of the medium. The embryonic development rate of the blastocyst and hatched blastocyst was higher, but not significantly so, when 0.4% MEM vitamins were supplemented to the in vitro maturation medium of the porcine oocyte. Interestingly, the total number of cells in the blastocyst was significantly higher in the in vitro maturation MEM vitamins supplemented group compared to either the untreated group or the group which had MEM vitamins supplemented to both in vitro maturation and in vitro culture medium simultaneously (p<0.05). Therefore, the supplementation of 0.4% MEM vitamins to the in vitro mature medium has a beneficial effect on the embryonic development of in vitro produced blastocysts derived from the immature porcine oocytes.

• 한국작물학회지
• /
• 제39권6호
• /
• pp.571-576
• /
• 1994

본실험은 대맥의 미숙배 배양을 통한 육종기간을 단축하기 위하여 미숙배 배양시 식물체 유도, 생육 및 출수에 영향을 미치는 생장조절물질, 배의 성숙정도 및 저온처리효과를 구명하고자 실시하였으며 그 결과는 다음과 같다. 1. 미숙배 배양으로부터 줄기의 유도에는 생장조절물질이 첨가된 기본배지도 효과적이었으며, Kinetin 0.5mg/1와 $GA_3$ 5mg/l를 처리한 것 이 좋았으나 Kinetin농도가 높을 때에는 shoot 유도가 감소되었다. 2. 지베렐린을 1mg/l와 5mg/l 처리한 배지에서 초장, root길이, root수 등이 좋았으나 Kinetin이 처리된배지에서는 생육이 억제되었으며, 특히 뿌리의 생육을 억제하였다. 3. 유도된 식물체의 토양생존율은 Kinetin 5mg/l 처리시에 생존율이 가장 낮았고 무처리에서 가장 높았다. 4. 파성정도가 IV인 올보리의 20일배를 4주저온처리 하였을 때에 출수율이 높았고 출수하는데 소요되는 기간도 짧았다. 5. 저온처리후 Kinetin 5mg/l 처리한 것은 초장, root수, shoot수가 적은 반면 $GA_3$ 5mg/l 처리한 것은 생육이 좋았다. 6. 올보리는 생장조절물질과 저온처리에 따라 출수율에 차이를 보였으며 $GA_3$를 1mg/l처리하고 28일 저온처리하였을때 출수율이 좋았으며, 출수에 소요되는 기간이 짧았다.

• Journal of Plant Biotechnology
• /
• 제6권4호
• /
• pp.213-219
• /
• 2004

Zygotic embryos were excised from immature and mature seeds of the Himalayan yew, Taxus wallichiana. The embryos germinated precociously when kept in darkness for 5 weeks and developed into full seedlings within 10-12 weeks. The highest rate of embryo germination ($81\%$) was obtained in modified Lloyd & McCown' s woody plant medium containing macro and micronutrients at half strength supplemented with $1\%$ activated charcoal, which supported both the best embryonic growth ($43\%$) and seedling development ($32\%$). However, the supplementation of basal media with kinetin, thidiazuron, 6-benzyl aminopurine or $GA_3$ had no effect on the germination of the embryos. The embryos derived from immature seeds germinated but the frequency of embryonic growth was better in mature seeds. Stratification of seeds effected precocious germination of embryos. Seeds kept at $4^{\circ}C$ for 1 week germinated earlier and at a higher frequency irrespective of the stage of seed maturity, while the germination rate declined with prolonged cold treatment for 1 month at that same temperature. Analysis of taxanes in germinating seedlings revealed that root tissues contained high levels of taxol, 10-deacetyl-baccatin ill and baccatin ill as compared to shoots. Thus embryo culture technique appears to overcome the lengthy dormancy requirement of T. wallichiana seeds.