• Title/Summary/Keyword: 분화율

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Plant Regeneration from Mature Seed-Derived Callus in Bermudagrass (Cynodon dactylon) (난지형 목초 버뮤다그라스의 종자유래 캘러스로부터 식물체 재분화)

  • Lee, Ki-Won;Park, Hyung-Soo;Choi, Gi-Jun;Kim, Ki-Yong;Ji, Hee-Chung;Kim, Kyung-Hee;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.3
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    • pp.223-228
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    • 2011
  • The present study was conducted to determine the optimum in vitro culture condition for callus induction and plant regeneration from mature seeds of bermudagrass (Cynodon dactylon cv. Common). It was revealed that mature seeds cultured on MS medium supplemented with 2 mg/L 2,4-D, 0.5 g/L proline, 0.5 g/L casamino acid and 3 g/L Gelrite under light condition produced the highest percentage of callus formation (39.2%). The most suitable medium for plant regeneration from dehydrated calli was MS agar medium supplemented with 0.5 mg/L 2,4-D, 2 mg/L BA, 0.5 g/L proline, 0.5 g/L casamino acid 3 g/L Gelrite which induced the highest percentage of calli forming shoots (57.7%). The frequency of callus induction and plant regeneration were the highest on sucrose, followed by maltose. The shoots were rooted at the highest rate (100%) when transferred onto 1/2 MS medium. Regenerated plants were morphologically uniform with normal growth pattern.

Plant Regeneration via Adventitious Shoot Formation from Hypocotyl Explants of Groundcherry (Physalis angulata L.) (땅꽈리(Physalis angulata L.) 하배축 절편으로부터 신초 형성을 통한 식물체 재분화)

  • Koh, Seok Chan
    • Korean Journal of Plant Resources
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    • v.35 no.4
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    • pp.502-507
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    • 2022
  • In the present study, plant regeneration through adventitious shoot formation from hypocotyl segments of in vitro seedlings of groundcherry (Physalis angulata L.) was investigated to determine the optimum culture conditions for highly efficient regeneration of the species. Adventitious shoots in hypocotyl segments were efficiently induced on MS media with low concentrations of BAP, specifically, with 0.5-1.0 mg/L BAP singly or in combination with 0.1-0.5 mg/L NAA. The 1.0 mg/L BAP single treatment was most effective for forming multiple adventitious shoots. When the induced shoots were transferred to the root induction media, low concentrations of NAA, IBA, and IAA enhanced the development of adventitious roots from adventitious shoots, suggesting that low concentrations of auxins were optimal for producing regenerated plantlets. The number of roots per shoot was large (> 2.0), and the root length exceeded 8.0 cm. In particular, the development and the overall shape of the roots were ideal. Furthermore, the number and length of shoots exceeded 2 and 6.0 cm, respectively. When the regenerated plantlets were transferred to compost soil, the root and shoot systems had developed well to the point that all of the regenerated plantlets acclimated successfully, resulting in normal morphology and growth characteristics, similar to those of the mother plant. Therefore, plant regeneration via adventitious shoot formation is expected to be one of the main methods for producing groundcherry on a large scale for a stable supply of the raw materials.

Regulation of Blastocyst Differentiation by the Serial Exposure of Conconavalin A and $PGE_2$ (Concanavalin A와 $PGE_2$의 순차적 노출에 의한 포배의 분화 조절)

  • Cheon, Yong Pil
    • Development and Reproduction
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    • v.12 no.3
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    • pp.267-274
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    • 2008
  • Differentiation of blastocyst is critical step for implantation and is under the control of regulation factors originated from embryo or reproductive tracts. The sequential communication with those factors is suspected as critical events for differentiation. It has been suggested that intracellular signaling pathways activated by calcium is essential in differentiation of blastocyst. Previously, it was known that concanavalin A (Con A) increase the levels of free calcium in blastocyst stage. However, Con A can not accelerate the hatching, although heparin-binding epidermal growth factor-like growth factor (HB-EGF), a modulator of calcium level, accelerate the hatching of blastocyst. In this study, it was investigated whether Con A or prostaglandin $E_2$ ($PGE_2$) can modulate the differentiation of blastocyst. Con A accelerated the expansion of blastocyst in both 1 hr pulse treatment group and continuous treatment group. However, Con A significantly suppressed the hatching in both groups. The inhibition was significantly strong in continuous treatment group compared with 1 hr pulse treatment group. On the other hand, $PGE_2$ induced the increase the free calcium level, but did not accelerate the expansion. In addition $10{\mu}m\;PGE_2$ inhibited hatching. However, $PGE_2$ could accelerate hatching in Con A pretreated blastocyst. $PGE_2$ also caused the increase of free calcium level in Con A pretreated blastocyst. From these results, it is suggested that changes of the free calcium level induce a different calcium-mediated signaling pathways. In addition, sequential stimulation by signal molecules may triggers the cellular mechanisms for the differentiation of blastocyst.

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Novel Heptapeptide Binds to the Lgr5 Induces Activation of Human Hair Follicle Cells and Differentiation of Human Hair Follicle Bulge Stem Cells (Lgr5와 결합하는 신규 헵타펩타이드를 이용한 인체 모낭 세포의 활성과 모낭줄기세포 분화 유도)

  • Min Woong Kim;Eung Ji Lee;Ha-Na Gil;Yong Ji Chung;Eun Mi Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.1
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    • pp.75-85
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    • 2023
  • This study was conducted to assess the effect of heptapeptide, composed of seven amino acids, on the activation of human hair cells isolated from human hair follicles. We have confirmed that the heptapeptide could bind to Lgr5 from the results of surface plasmon resonance (SPR) analysis. Heptapeptide enhanced the proliferation of human hair follicle dermal papilla cells (HHFDPCs) in a dose dependent manner. It induced the protein level of nuclear β-catenin, and the expressions of β-catenin downstream target genes, including LEF1, Cyc-D1 and c-Myc, in HHFDPCs. Heptapeptide significantly induced the phosphorylation of Akt and ERK, and the mRNA expressions of growth factors, including hepatocyte growth factor (HGF), keratinocyte growth factor (KGF) and vascular endothelial growth factor (VEGF), in HHFDPCs. In addition, heptapeptide significantly increased mRNA expression levels of differentiation-related transcription factors of human hair germinal matrix cells (HHGMCs) and differentiation markers of human hair outer root sheath cells (HHORSCs). Additionally, we investigated the effect of heptapeptide on human hair follicle stem cells (HHFSCs) differentiation and found that the heptapeptide reduced the mRNA and protein levels of stem cell markers, while it increased those levels of differentiation markers. These results have indicated that the heptapeptide promotes proliferation or differentiation of various types of hair follicle constituent cells through the induction of Wnt/β-catenin signaling. From the results, we have suggested that the heptapeptide in this study could be applied as a new functional material for the improvement of hair growth and alopecia.

Comparison of Regeneration Conditions in Seven Pepper (Capsicum annuum L.) Varieties (7종의 고추(Capsicum annuum L.) 재분화 조건 비교)

  • Min-Su Kim;Yun-Jeong Han;Sharanya Tripathi;Jinwoo Kwak;Jin-Kyung Kwon;Byoung-Cheorl Kang;Jeong-Il Kim
    • Korean Journal of Plant Resources
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    • v.36 no.5
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    • pp.527-539
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    • 2023
  • Pepper (Capsicum annuum L.) is an important vegetable and spice crop that has been cultivated worldwide. Pepper fruits have unique taste and aroma, providing a variety of antioxidants and compounds important for human health, which makes a high economic value. In addition, there is a high demand for new pepper varieties, according to consumer's preference. However, pepper is a recalcitrant plant for in vitro tissue and organ differentiation and plant regeneration, which makes it difficult to develop demanded varieties using newly developed technologies such as genetic engineering and gene editing. In this study, tissue culture and regeneration conditions were investigated using seven pepper varieties that were obtained from the core-collection of Seoul National University. We observed callus and bud induction and shoot formation using several media composition composed of different cytokinins and auxin concentrations. As a result, it was found that there were differences in callus induction and shoot formation of each variety depending on the hormone composition, and the highest regeneration was shown when the medium containing Zeatin Riboside and the petioles of seedlings were used. In particular, out of seven pepper varieties, CMV980 exhibited a higher regeneration efficiency (approximately 48%) than other varieties, followed by Yuwolcho. Therefore, this study provides CMV980 and Yuwolcho as good candidates that can be used for pepper transformation, which might contribute to the development of various varieties through gene editing technology in the future.

Effect of Plant Growth Regulators on Bulblet Formation and Plant Regeneration in Fritillaria thunbergii Miq. (패모조직배양에서 생장조절 물질이 자구형성 및 식물체 재생에 미치는 영향)

  • Park, Chul-Hyoung;Ryu, Jeom-Ho;Han, Kwang-Soo;Doo, Hong-Soo;Choi, Sun-Young
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.2
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    • pp.119-125
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    • 1996
  • To improve the efficiency of mass propagation in vitro of Fritillaria thunbergii, bulb scale and nodes were cultured in LS medium supplemented with the combination of 2, 4-D and kinetin or NAA and BA. The number and size of bulb, the number of adventitious shoot, the ratio of callus formation, rooting, and the effects of light and dark on the culture, plant regeneration from calli, and the gelling substances were investigated. The combination of 2, 4-D and kinetin in media was more effective than the media of NAA and BA for the bulblet formation. The media supplemented with 2 mg/L 2, 4-D and 1 mg/L kinetin, $1{\sim}2\;mg/L$ 2, 4-D without kinetin and $1{\sim}3\;mg/L$ BAA only were effective in the adventitious shoot development. Callus formation and root formation, respectively were effective in the medium supplemented with 2mg/L 2, 4-D and 1mg/L kinetin. In bulb formation, the medium with 5 mg/L kinetin was effective, and the most of bulbs were formed from the axillary bud of node part. In bulb formation, shoot growth, callus and root formation, the light culture for 16 hours per day was better than that in the dark culture. Bulb was nicely formed in the medium with 0. 2 mg/L 2, 4-D, 1 mg/L kinetin. The medium without hormone was most effective for plant regeneration. The phytagel was more effective than agar in the medium as the gelling agent.

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Plant Regeneration and Expression of Mouse Adenosine Deaminase Gene in Transgenic Hot Pepper (Capsicum annuum L.) Plants (형질전환된 고추( Capsicum annum L.) 식물체의 Mouse Adenosine Deaminas 유전자 발현)

  • 양덕춘;이계연;유영숙;최경화;임학태
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.37-41
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    • 1997
  • The in vitro regeneration and genetic transformation systems in hot pepper(Capsicum annuum L.) have not been routinely available, which has been a major limiting factor in the application of new genetic manipulations. An efficient procedure to regenerate whole pepper plants and to generate transgenic plants expressing a foreign gene was established. A relatively high frequency of plant regeneration was observed when hypocotyl and cotyledon explants were cultured on MS medium supplemented with NAA 0.1 mg/L plus zeatin 2.0 mg/L or IBA 10.0 mg/L plus BAP 1.0 mg/L. Addition of AgNO$_3$5 $\mu$M to these media improved the regeneration frequency up to 8%. For plant transformation, hypocotyl and cotyledon explants of hot pepper were precultured on shoot induction media without kanamycin added for 2 days, and then cocultured with Agrobacterium tumefaciens pDY183 for 2 days. Putative transformants were obtained from selection media containing 100 mg/L kanamycin sulfate and 500 mg/L carbenicillin. Putatively selected transformants were confirmed by amplification of selectable marker genes (ADA and NPT II) by polymerase chain reacion. Successful transcripts of ADA gene were detected by Northern blot analysis. Enzyme activity of ADA was also examined by spectrophotometric analysis, and expression of ADA gene in hot pepper suggests the potential application of ADA gene as a selectable marker in plants.

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Induction of Embryogenic Callus and Plant Regeneration by Mature Embryo Culture of Onion (Allium cepa L.) (양파의 성숙배 배양을 통한 체세포배발생 캘러스 유기 및 식물체 재분화)

  • Cho Kwang-Soo;Hur Eun-Joo;Hong Su-Young;Moon Ji-Young
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.31-35
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    • 2005
  • To obtain regeneration system of onion, we analyzed the effects of 2,4-D and BA concentration on the embryogenic callus induction from mature embryos. The highest embryogenic callus induction ratio was shown on MS medium (Murashie and Skoog 1962) containing $2.5\;\cal{mg/L}\;or\;5\;\cal{mg/L}$ picloram after mature embryos were placed on medium. When induced callus were cultured on half strength of MS medium containing $1\;\cal{mg/L}$ Kinetin, the highest shoot formation ratio was observed on MS medium containing $1\;{mg/L}$ 2,4-D and $1\;{mg/L}$ BA. Embryogenic callus were cultured in MS liquid medium containing $1\;\ccal{mg/L}$ of 2,4-D and $1\;\cal{mg/L}$ BA. The suspension cultured cell clumps could be mass propagated. Embryogenic callus were friable, but non-embryogenic callus included a lot of moisture, hence the identification between embryogenic and non-embryogenic callus as easily achieved. When embryogenic callus as cultured on half strength of MS medium containing $1\;\cal{mg/L}$ Kinetin, shoots were induced. The whole plantlet was obtained on rooting medium containing $0.5\;\cal{mg/}$ of NAA.

Factors Affecting Anther Culture in Schizandra chinensis (오미자(五味子)의 약배양(葯培養)에 영향(影響)하는 요인(要因))

  • Lee, Joong-Ho;Lee, Seung-Yeob
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.1
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    • pp.14-20
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    • 1997
  • To increase the efficiency of the callus induction in anther culture of Schizandra chinensis, the effects of culture stage, low temperature pretreatment, growth regulators, sucrose and gelling agents were tested on Murashige and Skoog's medium. And the effects of ABA and $AgNO_3$ on organogenesis were investigated. The most suitable stage for anther culture was at the middle to late uninucleate stage, of which the flower size was $6.2{\pm}0.6{\times}4.0{\pm}0.4mm(Length{\times}width)$, and the frequency of callus induction was 13.3%. The effect of low-temperature pretreatment on callus induction was highest as 12.5% in the trearment for 8 days at $4^{\circ}C$. The combination of 1.0 mg/L 2, 4-D and 0.25 mg/L kinetin for callus induction was most effective as 8.3% among various media. The frequency of callus induction was excellent as 10.8% in 4% sucrose. Effect of gelling agents on callus induction was highest as 9.0% on 0.6% Gelrite medium. The prevention of callus browning was effective on the media supplemented with ABA and $AgNO_3$ and rooting was promoted on medium supplemented with 5 mg/L ABA. But shoot was not developed.

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Transformation of Populus tremuloides Using Agrobacterium rhizogenes (Agrobacterium rhizogenes를 이용한 Populus tremuloides의 형질전환)

  • So, In-Sup;U, Zang-Kual;Ko, Young-Hwan;Lee, Sun-Joo;Hackett, Wesley P.;Riu, Key-Zung
    • Applied Biological Chemistry
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    • v.38 no.2
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    • pp.123-128
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    • 1995
  • Several factors affecting on transformation efficiency were studied to establish a Agrobacterium rhizogenes mediated system for the transformation of Populus species, and We could obtaine tansgenic plantlets expressing the introduced gene. Leaf sections were more sensitive than stem sections to kanamycin and thought to be better material for transformant screening. The bacterial density did not affect on the efficiency of transformation over the range of $4{\times}10^5{\sim}7{\times}10^9\;cfu$. The optimum period for co-cultivation was one day or shorter. Both of the optimum concentrations of cefotaxime and ampicillin in the medium were $250\;{\mu}g/ml$ for elimination of bacteria from the inoculated leaf sections. The addition of acetosyringone in the bacterial culture medium increased transformation rate, and the highest rate was obtained at $50\;{\mu}M$ of acetosyringone. The transformed galls could be selectively induced and gown on the growth regulator-free medium or on the medium containing $100\;{\mu}g/ml$ or higher contrition of kanamycin. The roots were induced from the galls incited by A. rhizogenes within 3 weeks on the growth regulator-free medium as well as on the medium containing growth regulators. The plantlets were regenerated from the galls cultured for 6 weeks on the medium containing 0.05 mg/ml of NAA and 0.5 mg/ml of BA. The expressions of the introduced opine gene in the transformed galls and plantlets were confirmed by the analysis of agropine and mannopine.

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