• Journal of Plant Biotechnology
• /
• 제6권1호
• /
• pp.45-50
• /
• 2004

Somatic embryos were induced from immature cotyledons and cultured on a MS medium containing 40mg/L 2,4-D. The maximum induction of embryos was obtained from immature cotyledons in a size of 3-4mm, and the highest frequency was obtained in the induction medium at pH 7.0. For embryo development, embryogenic tissues were transferred to a MSM6AC and MSM6 media. Developing embryos were placed at 27$^{\circ}C$with dim light (20$\mu$$molm^{-2}$$s^{-1}$) provided by cool fluorescent tubes (3-D wavelength light is better than standard light). Somatic embryos were clearly developed from globular stage to cotyledonary stages. The color of embryo may be a useful parameter for estimation of embryo quality. When the embryo becomes mature, embryo will be ready for desiccation in order to induce roots and shoots of embryos.

• 한국작물학회지
• /
• 제56권3호
• /
• pp.279-283
• /
• 2011

The immature embryos during seed development were examined to predict the suitable embryos for an efficient regeneration system. Five spring wheat genotypes and five winter wheat genotypes were tested using immature embryos as explants. Spring wheat genotypes showed much higher levels of plant regeneration than those of winter wheat genotypes. The highest frequencies of embryogenesis and regeneration were obtained when embryos at 13-14 days after anthesis (DAA) were used as explant and decreased using embryos at 21-22 DAA during seed development. Significant differences were also found for callus induction and regeneration as affected by immature embryo size. The regeneration efficiency was drastically decreased in spring and winter wheat genotypes when embryos larger than 2.0 mm of length were used. The optimum developmental stage and embryo length for regeneration efficiency were at 13-14 DAA and 1.0-1.5 mm, respectively. The selection of suitable embryos for the high frequencies of embryogenesis and regeneration leads us to efficient genetic improvement of wheat.

• 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.

• 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.

• Journal of Plant Biotechnology
• /
• 제41권1호
• /
• pp.44-49
• /
• 2014

In this study, we investigated the effects of sodium hypochlorite (NaOCl) and red or blue light-emitting diode (LED) light on embryo swelling and germination of Oreorchis patens (Lindl.) Lindl. A method for determining the swelling and protocorm formation in O. patens seeds through in vitro examination of immature seeds was established. NaOCl treatment of immature seeds was found to significantly enhance the extent of embryo swelling and protocorm formation in immature zygote embryos compared to those in the untreated controls. Additionally, the effects of white fluorescent light, and red and blue LED lights on embryo swelling and protocorm formation in in vitro cultured seeds were examined and compared to the conditions with or without NaOCl treatment. The most suitable light for embryo swelling and protocorm formation was the red LED light.

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2003년도 제3회 발생공학 국제심포지움 및 학술대회
• /
• pp.113-113
• /
• 2003

• Journal of Plant Biotechnology
• /
• 제7권3호
• /
• pp.187-193
• /
• 2005

A protocol was developed for somatic embryogenesis with 100% induction rate from immature zygotic embryo axes of peanut (Arachis hypogaea L. cv. DRG-12) cultured on MS medium containing $18.09\;{\mu}M$ 2,4-D. The frequency of somatic embryogenesis (31.7%) as well as the number of somatic embryos induced per explant (6.6) decreased when the concentration of 2,4-D was increased to $72.4\;{\mu}M$. Morphologically abnormal somatic embryos were observed at a frequency of 43.3% on MS medium containing $72.4\;{\mu}M$ 2,4-D. Somatic embryos isolated from 30-day-old cultures of immature zygotic embryo axes exhibited precocious germination with varied responses when placed on MS basal medium with 3% sucrose. Maximum shoot induction (80.0%) was observed from somatic embryos isolated from 60-day-old cultures of immature zygotic embryo axes when placed as a clump rather than individually on MS medium supplemented with $26.63\;{\mu}M$ BA and $0.54\;{\mu}M$ NAA. Shoots developed from somatic embryos rooted with higher frequency (93.3%) on Blaydes' medium containing $5.4\;{\mu}M$ NAA.

• Plant Biotechnology Reports
• /
• 제4권2호
• /
• pp.139-148
• /
• 2010

Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.

• 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.

• 한국가축번식학회지
• /
• 제21권4호
• /
• pp.355-362
• /
• 1997

This study was undertaken in an effort to product embryos through in vitro maturation(IVM), in vitro fertilization(IVF) and in vitro culture(IVC) after cryopreservation of immature and mature porcine oocytes. The experiments were conducted to investigate IVM rate of oocytes frozen with 3 different cryoprotectants and to examine IVF and IVC of frozen-thawed oocytes. The CEI(cumulus cells expansion index) after IVM of frozen-thawed immature oocytes was higher in oocytes frozen with PG＋PEG(propylene glycol plus polyethylene glycol) than those frozen with single cryoprotectant and this index was almost 90% of unfrozen oocyte's index(2.39 vs. 2.66). The IVF rate of all frozen oocytes was very low(68% of unfrozen oocytes) and the IVF rate of frozen immature oocytes was slightly higher than that of frozen mature oocytes(39.0% vs. 34.4%), but polyspermic penetration was higher in frozen immature oocytes(21.9% vs. 19.1%). The cleavage rate after IVF of frozen-thawed oocytes was 9.3% for frozen mature oocytes and 11.3% for frozen immature oocytes and this rate was significantly lower(P<0.05) than that of control(60.7%). The development to 8-cell stage was greatly lower in frozen mature oocytes than in frozen immature oocytes. The results indicate that the use of PG plus PEG as cryoprotectant may be very effective for vitrification of porcine oocytes and the frozen-thawed immature porcine oocytes can be used fro in vitro embryo production based on IVM, IVF and IVC system.