• Title/Summary/Keyword: callus cultures

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Effect of exogeneous plant growth regulators on morphogenetic response in vitro by embryo and leaf cultures of Camellia sinensis(L.) O. Kuntze (차나무 잎과 배 배양에 있어서 식물 생장조절물질이 형태형성에 미치는 영향)

  • PARK, Young Goo;AHN, In-Suk;BOZHKOV Peter
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.129-135
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    • 1997
  • Morphogenetic responses were investigated by culturing embryo and leaf explants of Korean wild type tea plant, Camellia sinensis (L.) O. Kuntze. Induction of direct somatic embryogenesis as well as adventitious and/or axillary shoots was obtained from mature zygotic embryo cultures on Murashige and Skoog (MS) basal medium having 5 to $20\mu\textrm{M}$cytokinin a lone. Morphogenetic response was decreased dramatically by the addition of auxins tested. One hundred percent of induced and isolated shoots formed roots after four weeks of culture on half-strength MS or quarter-strength Schenk and Hildebrandt (SH) media supplemented with $10\mu\textrm{M}$indole-3-butyric acid (IBA). Immature zygotic embryos were shown to be a suitable explant for embryogenic callus formation in the presence of 2, 4-dichlorophenoxyacetic acid(2, 4-D) in basal medium. Mature zygotic embryo originated leaves were used to test their ability for mophogenesis by incorporating plant growth regulators such as IBA, naphthyl-1-acetic acid (NAA), and 6-benzylaminopurine (BAP). Apparently, the morphogenetic responses of the cultured explant sources on the types and/or levels of plant growth regulators tested were observed visually.

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Somatic Embryogenesis and Plant Regeneration from Embryogenic cell Suspension Cultures of Schisandra chinensis Baill

  • Li, Cheng Hao;Niu, YudA;Zhao, Bo;Ghimire, Bimal Kumar;Kil, Hyun-Young;Heo, Kwon;Kim, Myong-Jo;Eom, Seok-Hyun;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.346-351
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    • 2007
  • An efficient somatic embryogenesis and plant regeneration protocol was developed for Schisandra chinensis Baill, using embryogenic cell suspensions and optimized media conditions. Friable embryogenic callus was induced from cotyledonary leaf and hypocotyl explants of 7 days old seedlings on MS agar medium supplemented with 1.0 to $4.0\;mg\;l^{-1}$ of 2,4-dichlorophenoxyacetic acid (2,4-D). Fast growing and well dispersed embryogenic cell suspensions were developed within two months when embryogenic calli were transferred to MS liquid medium containing $1.0\;mg\;l^{-1}\;2,4-D$. One third strength of MS medium was the best for both overall growth and development of somatic embryos in liquid culture. Over 3400 viable somatic embryos were produced from each 150 ml flask with an initial cell density of 30 mg in 30 ml medium. Germinated somatic embryos developed in liquid medium converted into plantlets after transferred to half-strength MS semi-solid medium. Approximately 90% of the converted plantlets were successfully transplanted to soil and grew into fertile plants.

Plant Regeneration via Somatic Embryogenesis and Organogenesis from Peucedanum japonicum $T_HUNB$ (갯기름나물의 체세포배 분화와 기관 분화를 통한 재분화)

  • Kim, Ok-Tae;Kim, Kwang-Soo;An, Jun-Chul;Hwang, Baik
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.21-24
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    • 2001
  • Peucedanum japonicum $T_{HUNB}$ used as a edible and medicinal plants was investigated for in uitro regeneration. Callus formation occurred on leaf and stem explant cultures and showed spontaneous embryogenic and organogenic capability on MS basal medium supplemented with 0.1~5 mg/L NAA and 0~10 mg/L BA in dark. The regeneration was highest on the condition supplemented with 2.5 mg/L NAA and 10 mg/L BA. Development of the somatic embryo progressed through the globular, heart-shaped, torpedo-shaped and cotyledonary stage, typical of zygotic embryos. When the first somatic embryos was cultured on the medium supplemented with 0.2 mg/L NAA, secondary somatic embryo were induced with higher frequency on the hypocotyl then on the cotyledon and root.t.

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Somatic Embryogenesis and Plant Regeneration in Suspension Cultures of Aralia elata S (두릅캘러스의 현탁배양에서 체세포배발생과 식물체 재분화)

  • 장한호;박철호;이윤수;신영범
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.167-171
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    • 1994
  • This study was carried out to investigate the possibility of plant regeneration through somatic embryogenesis in suspension culture of Aralia elata S. Callus was induced from the explants of leaf and petiole cultured in the MS media containing 2,4-D and TDZ. More embryogenic calli were formed from petiole and with combination treatment of 24-D and TDZ. The quarter strength MS medium was effective for increasing number of somatic embryos. Mannitol supplemented to the quarter strength MS medium, reduced somatic embryo formation but inositol increased. Normal plantlets(86%) were regenerated from mature somatic embryos in MS basal medium and 50% of those survied when transplanted to the vermiculite in greenhouse.

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Antitumor Activity of Cell Suspension Culture of Green Tea Seed (Camellia sinensis L.)

  • Choi, Jae-Hoon;Yoon, Sang-Kun;Lee, Keyong-Ho;Seo, Min-Soo;Kim, Doo-Hwan;Hong, Seung-Beom;Kim, Ji-Yeon;Paik, Hyun-Dong;Kim, Chang-Han
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.396-401
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    • 2006
  • The objective of this study was to investigate the antitumor activity of suspension cultures of tea callus cells grown in the presence of different concentrations of the growth regulator 2,4-dichlorophenoxy acetic acid (2,4-D) with or without light irradiation. The methanol and ethanol extracts of precipitated cells (MEP, EEP) exhibited stronger inhibitory effects on the growth of tumor cell lines than the water extract of precipitated cells (WEP) or the supernatant Compared to culture under dark conditions, exposure to light irradiation led to significantly higher antitumor activity. The MEP from light irradiated cells at $250{\mu}g/mL$ with 2.0mg/L 2,4-D displayed more than 64% growth inhibition of HEP-2 cells, whereas normal cells showed less than 25% growth inhibition. The some fractions of MEP obtained from Diaion HP-20 column chromatography displayed the majority of inhibitory activity against the HEP-2 cell line. These results show that 2,4-D, and light stimulated the synthesis of antitumor compounds.

Suspension Culture of Gardenia jasminoides Ellis Cell for Production of Yellow Pigment

  • Kim, Sang-Hwa;Park, Young-Goo;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.2
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    • pp.142-149
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    • 1991
  • Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.

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Characteristics of Hypovirulent Strains of Chestnut Blight Fungus, Cryphonectria parasitica, Isolated in Korea

  • Lee, Sang-Hyun;Moon, Byung-Ju;Lee, Jong-Kyu
    • Mycobiology
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    • v.34 no.2
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    • pp.61-66
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    • 2006
  • Chestnut blight disease caused by Cryphonectria parasitica is widely distributed throughout chestnut tree plantations in Korea. We surveyed 65 sites located at 9 provinces in South Korea, and isolated 248 virulent and 3 hypovirulent strains of chestnut blight fungus. Hypovirulent strains had dsRNA virus in the cytoplasm, which is one of the typical characteristics of hypovirulent strains. In addition, they showed more characteristics of hypovirulent strains, i.e., suppressed conidiation, reduced pigmentation in colony color, and reduced phenol oxidase activity as well as reduced pathogenicity. Hypovirulent strains, KCPH-22, KCPH-135 and KCPH-136, had a genomic dsRNA band with the molecular weight of 12.7 kb, which is the L-dsRNA of CHV1. They also had a 2.7 kb defective dsRNA band. Single conidia isolated from hypovirulent strains were cultured and various phenotypes and absence of dsRNA bands were obtained from single conidial cultures, which means that hypovirulence transmission is unstable in asexual reproduction and variations in viral heredity by asexual reproduction. Biocontrol trial using hypovirulent strains was also carried out in the chestnut tree plantations, and canker expansion in the treated trees was stopped and healed by callus formation at the margin of the canker. These results show the potentials in successful biocontrol of chestnut blight if the vegetatively compatible hypovirulent strains could be directly used around the canker formed by compatible virulent strains.

Relationship of Cotyledon Number with Procambium Differentiation in Somatic Embryogenesis of Codonopsis lanceolata L. (더덕의 체세포배발생에서 전형층분화와 자엽수의 관계)

  • Choi Pil-Son;Soh Woong-Young;Cho Duck-Yee;Liu Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.135-138
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    • 2005
  • Embryogenic callus was obtained from cotyledonary explants of Codonopsis lanceloata on Murashige & Skoog's medium supplemented with 1 mg/L 2,4-D. Suspension cultures of the embryogenic calli were grown on a shaker at 100 strokes/min, and then the calli were subcultured for 2 weeks in 2,4-D-free medium to produce somatic embryo. In somatic embryos at the globular stage, cotyledon initials began to differentiate themselves in the near distal end of the procambial strand. Dicotyledons, tricotyledon, tetracotyledon and fused cotyledon were differentiated from the distal ends of two, three, four and circular procambial strands, respectively. Nearly circular procambial strand in lower hypocotyls were independently differentiated into two, three, four procambial tissues at cotyledonary node and cotyledons to form somatic embryos with dicotyledon, tricotyledon, tetracotyledon. If the distal subepidermal cells of globular embryo exclusively became cotyledon initials, the torpedo or cotyledonary embryo was characterized by somatic embryos with fused cotyledon.

The Effects of Ammonium Citrate and Ammonium Succinate on the Growth of Cells and Nitrogen Absorption in Korean Ginseng Suspension Cultures (고려인삼(Panax ginseng C.A. Meyer) 배양세포의 생육 및 질소 흡수에 미치는 Ammonium Citrate와 Ammonium Succinate의 영향)

  • 김홍성;김명원;소상섭;강영희
    • Journal of Plant Biology
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    • v.24 no.1
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    • pp.13-19
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    • 1981
  • In order to investigate the effects of ammonium citrate and ammonium succinate on the growth and absorption of nitrogen compounds supplied in the medium, Korean ginseng (Panax ginseng C. A. Meyer) calli were suspension cultured in MS medium with various concentrations of ammonium citrate and ammonium succinate. When Korean ginseng calli were cultured with 10 mM ammonium citrate, 10 mM ammonium succinate, and 10 mM ammonium nitrate (control) in MS media as the nitrogen sources, the growth, $NO_3$-N absorption and total nitrogen content of the Korean ginseng cells were greatest in the ammonium citrate and ammonium succinate concentrations. When Korean ginseng calli were cultured with 5 mM ammonium citrate and 5 mM ammonium succinate, the growth and nitrogen content were superior to those of the control: however, $NO_3$-N and $NH_4$-N absorptions were similar to those of the control. In conclusion, the 10 mM ammonium citrate and 10 mM ammonium succinate may be better able to facilitate the growth and $NO_3$

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An Effective Selection of PAT Gene Transformed Populus alba $\{times}$ Populus glandulosa No.3 using Herbicide Basta Treatment (제초제 Basta를 이용한 Phosphinothricin Acetyltransferase 유전자로 형질전환된 현사시 3호의 효율적인 선발)

  • 오경은;문흥규;박재인;양덕춘
    • Korean Journal of Plant Resources
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    • v.17 no.1
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    • pp.28-33
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    • 2004
  • This study was conducted to simple transformants selection by herbicide Basta treatment after transformation with Agrobacteium tumefaciens MP90/PAT in hybrid poplar(Populus alba ${\times}$ P. glandulosa No. 3). In preliminary study, we determined that the lethal concentration of herbicide Basta was 1.0mg/L in callus culture, adventitious bud formation and axillary bud elongation experiment. By the treatment of 1.0mg/L Basta, we could be selected the transformed shoots effectively from the various cultures. In addition, the treatment was useful on selection of transformants which are growing in soil pot. Finally, we also confirmed the transformation by PAT assay, Above results show that the herbicide Basta treatment and PAT assay can be a very simple and effective method for the identification of PAT gene transformed hybrid poplar.