• Title/Summary/Keyword: explants

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Callus and Micro-Crown Bud Formation in Vitro from Leaf Explant of Yacon (Polymnia sonchifolia Poeppig & Endlicher) (야콘 (Polymnia sonchifolia Poeppig & Endlicher) 잎의 절편체로부터 캘러스 및 기내 소관아 형성)

  • 두홍수;권태호;박철형;류점호
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.101-107
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    • 2000
  • The explants of yacon (Polymnia sonchifolia Poeppig & Endlicher) were cultured to invest th8e dedifferentiation condition, and formative callus from leaf was cultured to find the regeneration and micro-crown bud formation. Basal MS medium was more effective to form callus than 1/2 MS and B$_{5}$ medium. Calli formations from leaf, petiole and lateral bud were more effective on MS medium supplemented with 1.0, 2.0 mg/L 2,4-D and 0.2, 0.4 mg/L kinetin or BA than 1.0, 2.0 mg/L NAA and 0.2, 0.4 mg/L kinetin or BA. Formative callus from leaf was proliferated about 70% on medium supplemented with 1.0 mg/L BA. When callus was proliferated, 63% regeneration rate was shown on medium supplemented with 1.0, 2.0 mg/L BA in case of subculture for 3~4 months but was not shown on medium supplemented with 1.0, 2.0 mg/L kinetin. Micro-crown bud formed as addition of BA at 3~4 months after callus culture and then was obtained many at 5~6 months, it was most formed about 82% on medium supplemented with 5 mg/L BA. Rate of micro-crown bud formation was increased as more over 5 mg/L BA concentration, when this time, however, shoot had thick leaves and short internodes, and then withered before long, Micro-crown bud was formed about 88.0% on medium supplemented with 5% sucrose, that was more increased 28% than with 3% sucrose. The buds of crown bud between harvested in field and formed in vitro were difference only in size, but both were similar in shape according to histological view.

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Effect of 2,4-Dichorophenoxyacetic Acid on Adventitious Root Formation from Callus Bupleurum falcatum L. and Its Histological Observation (시호(Bupleurum falcatum L.)의 캘러스로부터 부정근 분화에 미치는 2,4-Dichlorophenoxyacetic Acid의 영향과 분화의 해부학적 고찰)

  • 배형화;조덕이;김성길;소웅영;성낙선
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.41-46
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    • 1994
  • Calli were induced from leaf explants of B.falcatum, and selected cell clumps of the calli (900-1, 000${\mu}{\textrm}{m}$) were cultured on MS medium supplemented with 0.1, 0.5, 1.0 or 2.0 mg/L 2, 4-D for 7 days, respectively: The clumps were subsequently transferred onto MS basal medium and subcultured for four weeks. In order to investigate the effect of 2, 4-D pretreatment, the selected clumps were cultured on MS medium supplemented with 0.1 mg/L 2, 4-D for 24, 48, 72, 96, 120 or 144 hours and then transferred to liquid MS basal medium, wherein they were cultured for 4 weeks. Histological observation showed that root initial cells were developed from cells on the surface of clumps or from cells in the inner region. Clumps on the basal medium produced mot within 5 days of culture. The rate of prutruding time was inversely proportional to the concentration of 2, 4-D. The number of adventitious roots per clump preheated with 0.1 mg/L 2, 4-D was an average of 5.2, which was the highest level. On MS medium as control, the clumps formed 3.3 adventitious roots each. As tile concentration of 2, 4-D increased, the number of adventitious roots were declined accordingly: The number of adventitious roots as the period of pretreatment increased upto 120 h.

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Plant Regeneration from Hypocotyl Explants of Several Species of Lycopersicon (토마토속 식물의 배축절편 배양에 의한 식물체 재분화)

  • 임학태;이건섭;용영록;송융남;김종화
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.137-143
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    • 1994
  • In an attempt to optimize the in vitro-regeneration conditions necessary for the genetic manipulation of tomato species, we examined several hybrid lines and wild species (peruvianum, pimpinellifolium, glandulosum) of Lycopersicon for. their, different regeneration ability. The basal medium used for callus growth and organogenesis was MSB (MS + B5) supplemented with three combinations of TDZ (Thidiazuron) 0.5mg/L+NAA 0.5mg/L, BA 2.0mg/L+NAA0.05 mg/L, and zeatin 3.0 mg/L + IAA 0.02 mg/L. In the genotype of Lycopenicon grandulosum, combination of TDZ and NAA was more effective in inducing shoot and root differentiation than those of BA and NAA or zeatin and IAA. When all genotypes tested were considered, however combination of zeatin and IAA was shown to be the best in shoot regeneration. Result indicate that callus and organogenesis of Lycopenicon species are dependent upon the hormone types and plant genotypes, but MSB medium with zeatin 3.0 mg/L + IAA 0.02 mg/L maybe appropriate for genotype-independent plant regeneration system of Lycopercicon species. We also tried TDZ as a cytokinin source in tomato tissue culture and found it highly significant in tomato regeneration system.

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Stable Transformation via Callus Formation and Rhizogenesis from the Cultures of Hypocotyl Explant of Chinese Cabbage (배추의 배축절편으로부터 캘러스와 뿌리 발생을 통한 안정적 형질전환)

  • Cho, Mi-Ae;Kim, Choon-Ae;Min, Sung-Ran;Ko, Suck-Min;Liu, Jang-Ryol;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.139-144
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    • 2007
  • Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.

Efficient shoot regeneration using cotyledon explants and Agrobacterium tumefaciens - mediated transformation of bottle gourd (Lagenaria sicraria Standl.) (박 자엽조직을 이용한 효율적인 식물체 분화와 Agrobacterium에 의한 형질전환)

  • Kim, Soo-Yun;Ahn, Yul-Kyun;Huh, Yun-Chan;Lee, Hye-Eun;Kim, Do-Sun
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.140-145
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    • 2014
  • This study were carried out for selection of proper transformation variety and development of efficient regeneration and transformation methods. The number of shoot in commercial varieties of gourd plant were 0 ~ 7.3. and fusarium wilt resistant pure lines were 2.0 ~ 6.5 per dish containing on MS medium supplemented with 3 mg/L BA. The shoot regeneration frequency of fusarium wilt resistant pure lines were wide variation on the deviation. The expression of GFP was high 67% and 100% at the co-cultivation with Agrobacterium. The effective shoot regeneration plant hormone were combination BA and 2,4-D. The number and elongation condition of shoot was good after 4 weeks change with MS medium supplemented with 1 mg/L BA. Effective callus production plant hormone were combination of 3 mg/L BA and 0.1 mg/L 2.4-D.

In vitro plantlets regeneration by multi-shoots induction and rooting in Chamaecyparis obtusa (편백의 다신초 유도 및 발근을 통한 식물체 재분화)

  • Kim, Ji Ah;Lee, Na-Nyum;Kim, Yong Wook
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.303-309
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    • 2019
  • A protocol for the in vitro propagation of Chamaecyparis obtusa was established in the present study. Multi-shoots were initiated from apical shoot explants from germinants after 10 weeks of culture on Litvay medium (LM) supplemented with different concentrations of cytokinin. The effects of pre-treatment with high concentrations of cytokinin and varying concentrations (0.2 to 5.0 mg/L) of zeatin on in vitro shoot elongation and shoot multiplication were investigated. Optimal shoot growth was achieved on LM medium, with over 10-mm shoots after 10 weeks of culture. In the anti-browning tests, ethanesulfonic acid triggered the least browning in the shoot tips. The highest multi-shoot induction was observed in the 0.5-mg/L zeatin treatments, which yielded 80% induction of shoots after 10 weeks of culture, and maximum shoot elongation was observed in the LM basal medium without the hormone. The highest rooting rates were 65% under 0.2 mg/L indole-3-butyric acid.

Improvement of Black Locust(Robinia pseudoacacia L.) Through Tissue Culture. I. Micropropagation and Somatic Embryogenesis (조직배양에 의한 아까시나무(Robinia pseudoacacia L.)의 개량 I. 대량증식과 체세포배 발생)

  • Woo, Jong Ho;Choi, Myung Suk;Joung, Eun Yi;Chung, Won Il;Jo, Jin Ki;Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.84 no.1
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    • pp.41-47
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    • 1995
  • A micropropagation system for black Locust(Robinia pseudoacacia) was established by using shoots and pin-punctured leaves of in vitro germinated seedlings. The greatest number of shoots (an average of 10.5 shoots) was obtained when shoot tips were cultured on MS medium supplemented with 1.0 mg/l BAP and 0.01 mg/l NAA. When pin-punctured leaf explants were cultured on the same medium, mean number of 13.5 shoots were produced. Shoot growth was accelerated by adding 50 mg/l of silver nitrate ($AgNO_3$), an anti-ethylene compound to the culture medium. Each shoot was excised from the mass and transferred onto half strength MS medium for rooting. Zygotic embryos at different developmental stages were cultured on LS medium supplemented with various growth regulators to induce somatic embryos. When cultured on LS medium with 1.0 mg/l 2,4-D. 14.3% of the zygotic embryos induced somatic embryos. Upon transfer onto the basal medium, somatic embryos sporadically converted into plantlets.

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Transient Expression of β-gulucuronidase (GUS) gene in Immature Ovules and Calli Derived from Cottonwood Species (Populus deltoides) by Microprojectile Bombardment (포플러의 미성숙(未成熟) 배(胚)와 캘러스에서 유전자총(遺傳子銃)에 의(依)한 GUS-gene의 일시적(一時的) 발현(發現))

  • Kang, Hoduck;Kang, Sang-Gu;Bae, Hanhong;Park, Kyo-Soo;Hall, Richard B.
    • Journal of Korean Society of Forest Science
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    • v.86 no.3
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    • pp.261-269
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    • 1997
  • Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and ${\beta}$-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-${\beta}$-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced about $25.75{\pm}2.77$(per ovule), $11.43{\pm}1.22$(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.

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Interleukin-$1{\beta}$ induces bone resorption by regulation of prostaglandin $E_2$ synthesis and plasminogen activator activity, and TGF-$\beta$ inhibits bone resorption of rat bone cells (쥐의 골세포에서 $PGE_2$ 합성과 plasminogen activator 활성 조절에 의한 IL-$1{\beta}$의 골 흡수유도와 TGF-$\beta$에 의한 골 흡수 억제 기전에 관한 연구)

  • Kim, Young-Hun;Lee, Young-Jun;Chung, Kyu-Rhim;Park, Young-Guk
    • The korean journal of orthodontics
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    • v.30 no.6 s.83
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    • pp.713-721
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    • 2000
  • Bone cells produce multiple growth factors and cytokines that have effects on bone metabolism and can be incorporated into the bone matrix. The present study was designed to extend these observations by examining the interactions between transforming growth factor-$\beta$(TGF-$\beta$) or interleukin-$1\beta$(rhIL-$1\beta$) and bone cells in a rat long bone culture model. IL-$1\beta$ regulates several activities of the osteoblast cells derived from rat long bone explants in vitro. IL-$1\beta$ stimulated cellular proliferation as well as the synthesis of prostaglandin $E_2$ and Plasminogen activator activity in the cultured cells in a dose-dependent manner. TGF-$\beta$ is present in the bone matrix and potentially released during bone resorption. TGF-$\beta$ reduced basal bone resorption and inhibited vitamin $D_3[1,25(OH)_2D_3]$-induced bone resorption in rat long bone cells. These results support the role of IL-$1\beta$ in the pathological modulation of bone cell metabolism, with regard to implication in the Pathogenesis of osteoporosis by IL-$1\beta$, and that TGF-$\beta$ positively inhibits the bone resorption.

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Chromosome Doubling of Allium wakegi Araki by In Vitro Cultures (기내배양에 의한 쪽파의 체세포 염색체 배가)

  • Yim, Sun-Hee;Ahn, Chang-Soon;Jeong, Chang-Nam;Han, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.259-264
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    • 2002
  • Induction of embryogenic callus from Allium wakegi Araki explants was promoted on medium containing 2,4-D, and production of abnormal embryos from the embryogenic callus increased as 2,4-D concentration was raised. Shoot tip was found to be the best explant source for embryogenic callus formation followed in the order by bulb scale and leaf section. Medium containing 0.09 M sucrose was effective for embryogenic callus production. The regenerants from embryogenic callus on medium containing 2,4-D and BA at different concentrations was consisted of diploids, tetraploids and a few mixaploids of 2n+4n, and their chromosomal aberration rate ranged from 8.0 to 33.3%. Frequency of chromosomal aberrants was the highest (18.7%) in the regenerants obtained from bulb scale-derived embryogenic callus among others. Plant regeneration rate was high (33.5%) from the shoot tip-derived embryogenic callus and the frequency of chromosomal aberrants was very low (7.0%). The plantlets regenerated on medium containing 0.26 M sucrose resulted in high chromosomal aberrants. The regenerants on medium containing sucrose at 0.09∼0.20 M produced chromosomal aberrants at around 15.2∼16.6%.