• Journal of Plant Biotechnology
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• 제50권
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• pp.267-274
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• 2023

All studies on date palm somatic embryogenesis have focused on germination in the presence of light while neglecting germination in darkness, which mimics the germination process of zygotic embryos within seeds. To improve the date palm micropropagation protocol, we investigated the effects of light and darkness incubation on the germination of indirect date palm somatic embryos and their subsequent conversion into plantlets. Darkness incubation emerged as a pivotal factor in the germination of indirect date palm somatic embryos and their successful conversion into plantlets. Darkness incubation significantly decreased the time required for the conversion of indirect somatic embryos into plantlets, halving the duration from 24 weeks to only 12 weeks. The micropropagation protocol was modified, consolidating the previous two distinct stages of germination and elongation under light incubation into a single stage under darkness incubation. These findings modified the protocol and significantly reduced the overall duration of the date palm micropropagation protocol.

• 한국동물생명공학회지
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• 제34권2호
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• pp.130-138
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• 2019

Although somatic cell nuclear transfer (SCNT)-derived embryonic stem cells (ESCs) in pigs have great potential, their use is limited because the establishment efficiency of ESCs is extremely low. Accordingly, we tried to develop in-vitro culture system stimulating production of SCNT blastocysts with high performance in the colony formation and formation of colonies derived from SCNT blastocysts for enhancing production efficiency of porcine ESCs. For these, SCNT blastocysts produced in various types of embryo culture medium were cultured in different ESC culture medium and optimal culture medium was determined by comparing colony formation efficiency. As the results, ICM of porcine SCNT blastocysts produced through sequential culture of porcine SCNT embryos in the modified porcine zygote medium (PZM)-5 and the PZM-5F showed the best formation efficiency of colonies in α-MEM-based medium. In conclusion, appropriate combination of the embryo culture medium and ESC culture medium will greatly contribute to successful establishment of ESCs derived from SCNT embryos.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.213-237
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• 1987

In vitro flowering system may minimize the confounded influence of non-floral meristem parts of plants in studying the relationship of a given treatment and flowering responses. We have induced flower buds from plantlets regenerated from zygotic embryo-derived somatic embryos of ginseng, which circumvented the normal 2-year juvenile period before flowering. The result suggests that the adulthood of ginseng root explants in the experiment previously conducted by Chang and Hsing (1980; Nature 284: 341-342) is not prerequired to flowering of plantlets regenerated through somatic embryogenesis. We have also induced flower buds from elongated axillary brandches from cotyledonary nodes by culturing ginseng zygotic embryos, seedlings, and excised cotyledonary nodes. It was found that 6-benzyladenine (BA) supplemented to the medium was essential for flowering, whereas abscisic acid (ABA) was inhibitory. Gibberellic acid(GA3) was also required for flowering when ABA was present with BA in the medium. The results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respective-ly, in the induction of flowering of ginseng. Tran Thanh Van (1980; Int. Rev. Cytol., Suppl. IIA: 175-194) has developed the "thin cell layer system" in which the induction of shoots, roots, or flower buds from epidermal layer explants were controlled by culture conditions and exogenous growth regulators in the medium, Utilizing the thin cell layer system, Meeks-Wagner et al. (1989; The Plant Cell 1: 25-35) have cloned genes specifically expressed during floral evocation. However, the system is too tedious for obtaining a sufficient amount of plant materials for biochmical and molecular biological studies of flowering. We have developed a garlic callus culture system and one obvious advantaging over the thin cell layer system is that an abundant cells committed to develope into flower buds proliferate. When the above cells were compared by two-dimensional gel electrophoresis with those which have just lost the competence for developing into flower buds, a few putative proteins specific to floral evocation were detected. The garlic callus culture system can be further explored for elucidation of the molecular biological mechanism of floral evocation and morphogenesis.hogenesis.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2003년도 춘계 학술발표대회
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• pp.61-62
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• 2003

Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

• Plant Biotechnology Reports
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• 제2권4호
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• pp.259-265
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• 2008

Picea koraiensis, called Korean spruce, is an evergreen tree and found mostly in northeast Asia. In this study, plant regeneration via somatic embryogenesis from open-pollinated immature zygotic embryos of nine geno-types of elite trees was established. Immature zygotic embryos were cultured onto RJW medium modified from 505 medium with $21.48{\mu}M$ NAA, $2.22{\mu}M$ BA, and $2.32{\mu}M$ KT. The average frequency for all nine genotypes was 74.2%. Embryogenic calluses of the nine genotypes of elite trees were subcultured on RJW basal medium containing $8.06{\mu}M$ NAA, $1.11{\mu}M$ BA, and $1.16{\mu}M$ kinetin. The calluses of three lines, $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$, were actively proliferated but others were not. Somatic embryogenesis was induced from the embryogenic callus in genotypes of $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$ on RJW medium with ABA and $60g\;l^{-1}$ sucrose. Cotyledonary somatic embryos were subjected to a drying process. The drying of embryos by uncapping the culture bottle for 5 days on a clean bench resulted in a high frequency of germination of somatic embryos (87% in RJW medium). However, plantlet conversion from germinated embryos was greatly reduced and the optimal medium for plant conversion was 1/2 WPM or 1/2 BMI medium. In conclusion, we have, for the first time, established a plant regeneration system via somatic embryogenesis in the Korean spruce, which can be applied for rapid micropropagation of elite trees.

• 식물조직배양학회지
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• 제22권4호
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• pp.189-194
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• 1995

유자의 배배양에 의한 캘러스의 유도는 미숙열매의 배로부터 가장 양호하게 유기되었다. 이들 캘러스는 3% sucrose 와 44.4 $\mu$M BA가 첨가된 1/2 MS배지에 치상하였을 때 가장 잘 유도되었으며 배양조건은 26$^{\circ}C$ 에서 2000 lx로 16 h의 광주기와 8 h의 암주기로 배양하였다. 이들 캘러스는 담황색으로 표면에 크고 작은 많은 돌기들을 가지고 있는 캘러스와 암갈색의 캘러스가 관찰되었다. 또한 캘러스 발달 단계와 기관분화의 정도에 따라서 세포배열 양상에 많은 차이가 있었는데, 배양 6주된 캘러스는 작은 세포들이 조밀하게 나타났으며 8주경부터 신장된 세포들이 분열하는 양상을 보여 주었으며 주된 캘러스 표면의 돌기들은 기관 발생의 초기에 구형의 세포돌기가 융기된 반면 주변의 세포들 은 액포화된 것으로 나타났다. 배양 16주 후에, 재분화된 shoot는 MS 배지에 이식하였을 때 발근이 되어 정상적으로 생장하였다. Shoo떼서는 유관속 형성층과 정유를 함유하고 있는 유낭의 초기 발생과정이 광학현미경으로 관찰되었다. 형성된 shoot를 화분에 이식하였을때 정상적으로 생장이 계속되는 것을 관찰하였다.

• Journal of Plant Biotechnology
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• 제35권1호
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• pp.47-53
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• 2008

겨우살이의 캘러스 형성 및 종자 발아 와 haustorium 발달에 요구되는 환경요인 및 생장조절제의 영향을 조사하였다. 아울러 종자의 인위부착을 통한 식물체 재생 체계를 확립하였다. 겨우살이 조직관찰 결과 종자에는 1개 또는 2개의 막대모양의 접합자배가 존재하며 종자의 외부에 위치한 유근 부위에 주로 세포분열이 왕성한 세포층이 존재하며 반대로 자엽은 종자 내부에 위치하며 상대적으로 분열능을 가진 세포수가 감소하였다. 겨우살이 종자 발아 와 haustorium 발달에 요구되는 여러 요인 중에서 광의 요구도가 절대적이었다. 본 실험에 조사된 다른 배양 환경요인 및 생장조절제들은 광의 효과를 대체하지 못하였으며 ethylene의 경우 발아촉진 효과가 3배 이상 증가함을 알 수 있었다. 여러 조직 중에서 오로지 겨우살이 화아로부터 캘러스 형성이 가능하였으며 캘러스 형성빈도는 $0.1\;mgl^{-1}$ IAA가 첨가된 B5 배지에 배양 시 27.3%로 가장 높았다. 본 연구에서 확립된 겨우살이 캘러스 배양 및 종자 인위 부착을 통한 식물체 재생체계는 겨우살이의 유용물질 생산 연구 및 대량증식 연구분야에 활용이 가능할 것으로 기대된다.

• 한국자원식물학회지
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• 제18권1호
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• pp.123-130
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• 2005

인삼포장에서 염류의 집적은 우량인삼의 생산에 많은 장애요인이 되고 있다. 본 연구는 인삼종자의 NaCl에 대한 반응정도를 조사하고, 또한 체세포배의 발생시 및 인삼모상근에서 사포닌의 생성에 미치는 반응을 조사하였다. 인삼 접합자 배를NaCl이 첨가되지 않은 배지에 접종한 결과 $84.5\%$가 발아되었으며 0.1M에서는 $3\%$ 만이 발아되었고, 0.2M이상의 처리구에서는 전혀 발아가 되지 않았다. 또한 체세포배 발생 은 미숙배를 이용할 경우 NaCl 무첨가배지에서 가장 양호하였으며 농도가 증가할수록 감소하는 경향을 보였다. 반면에 성숙배에서 는 NaCl 무첨가배지보다 0.05M첨가배지에서 더 양호한 경향을 보였다. 뿌리 상태인 인삼의 모상근의 경우에는 광상태에서 NaCl를 첨가한 배지에 배양시 0.04M에서부터 안토시안이 형성되기 시작하여 0.08M에서는 눈으로 확인이 가능할 정도의 많은 량이 형성되었다. 안토시안을 함유한 세포주는 다른 세포주에 비해 crude saponin뿐만 아니라 total ginsenoside의 함량도 높은 경향을 보였으며 panaxatriol보다 panaxadiol이 2배 이상 많았다.

• 화훼연구
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• 제17권4호
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• pp.221-225
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• 2009

오리엔탈나리 (Lilium spp.)를 모본으로 하여 환경적응성이 우수한 종간잡종 나리를 육성하기 위하여 종간 교잡을 실시하였다. 오리엔탈 ${\times}$ 아시아틱 종간교잡은 화주절단수분법에 의해 수분하였지만 평균 29%왜 낮은 결실률을 나타냈다. 오리엔탈 나리 'Rodolfa' ${\times}$ L. asiatic 'Toronto'는 대조구에 비하여 1/4 MS나 1/6 MS배지에서 76.6~78.3%로 발아율이 높았으나, 오리엔탈 나리 'Farolito' ${\times}$ L. asiatic 'Buff Pixie'는 MS와 1/2 MS배지에서 발아율이 86~90% 크게 향상 되었다. 그러나 'Snow Cristal' ${\times}$ Lilium asiatic 'Royal Trinity'는 모든 처리구에서 100%의 발아율을 나타냈다. 종간 잡종배는 배지에 GA $0.5{\sim}1.0mg{\cdot}L^{-1}$ 첨가에 의해서 발아율이 약간 향상되었다. 오리엔탈 나리 'Farolito' ${\times}$ L. asiatic 'Buff Pixie'와 'Belcanto' ${\times}$ L. callosum는 $0.01mg{\cdot}L^{-1}$ NAA와 $0.01mg{\cdot}L^{-1}$ kinetin이 첨가된 MS 배지에서 66.6%와 45.2%로 가장 높은 발아율을 나타냈다.

• 식물조직배양학회지
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• 제28권1호
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• pp.21-24
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• 2001

식용 및 약용으로 사용되는 갯기름나물의 기내배양을 통한 재분화율을 조사하였다. 캘러스는 잎과 엽병의 절편체에서 발생하였고 0∼5 mg/L의 NAA와 0∼10 mg/L BA가 첨가된 MS 기본배지에 치상하여 암배양 결과 자발적인 배 분화와 기관 분화를 보여주었으며 2.5 mg/L NAA와 10 mg/L BA의 조건에서 재분화율이 가장 높았다. 체세포 배의 발생양상은 구형, 심장형, 어뢰형을 거쳐 자엽 단계로 발달하여 전형적인 접합자 배의 모습으로 발달하였다. 배 발생한 일차 체세포배를 0.2 mg/L의 NAA가 첨가된 배지에서 배양하였을 때, 이차 체세포배가 발생하였으며 주로 일차 체세포배의 배축과 하배축에서 많이 발생하였고 그 다음으로 자엽이나 뿌리 부분에서 발생하였다.