• Title/Summary/Keyword: plant tissue

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Cloning and Characterization of Homeodomain-Zip Gene, Phc5 in Embryogenic Callus derived from Pimpinella brachycarpa Suspension Cultured Cells (참나물 현탁배양세포 유래 배발생캘러스에서 HD-Zip 유전자, Phc5의 클로닝과 특성)

  • 손수인;김준철
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.121-126
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    • 1999
  • Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

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Effect of Inorganic Salts in MS Medium, Sucrose, and Activated Charcoal on Bulblet Formation from in Vitro Bulbscales in Lilium Oriental Hybrid 'Casa Blanca' (MS 배지 무기물, 당 및 활성탄의 농도가 Lilium Oriental Hybrid 'Casa Blanca'의 기내인편으로부터 자구형성에 미치는 영향)

  • 한봉희;예병우;구대회;고재영
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.103-107
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    • 1999
  • The effects of MS salt strength, sucrose, and cultural conditions on bulblet formation and growth were investigated to optimize the conditions for micropropagating bulblets from in vitro bulb scales of Lilium Oriental Hybrid 'Casa Blanca'. There was no difference on bulblet formation in the range of 1/2~2 $\times$ strength of MS salt, but it was inhibited remarkably in 3 $\times$ strength of MS salt. The growth of regenerated bulblets was most stimulated on MS basal medium. Favorable bulblet formation and its growth from bulb scales were achieved when grown on the media with 1 : 2 or 1 : 3 in the ratio of NH$_4$^+ : NO$_3$^- , as well as on MS basal medium (NH$_4$^+ : NO$_3$^- = about l : 2). Therefore, MS basal medium was very suitable for bulblet formation and growth from bulb scales. Bulblet formation was inhibited but its growth was stimulated with increase sucrose concentration in the medium. The growth of regenerated bulblets was very effective on the media with 9~12% sucrose. Addition of activated charcoal (AC) to the medium inhibited bulblet formation from bulb scales, but enhanced the growth of regenerated bulblets. Especially, the medium containing 1 g/L AC was most effective on the growth of bulblets. No difference was found on bulblet formation and growth from bulb scales under light and dark conditions. In vitro micropropagation of L. Oriental Hybrid 'Casa Blanca' was supposed very reasonable to enhance the growth of the bulblets after forming of bulblets from in vitro bulb scales, and then, subculture the bulb scales from the grown bulblets on MS medium with 9% sucrose and 1 g/L AC.

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Growth Acceleration and Acclimatization of In Vitro Plantlets derived from Apical Meristem of Sweet Potato (고구마의 경정조직 유래 기내 소식물체의 생장촉진과 순화)

  • ;;Shiro Higashi
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.115-119
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    • 1999
  • The single node cuttings of sweet potato (cv. Mokpo #29) plantlets maintained in vitro were cultured with (MF+) or without membrane filter (MF-) under photomixotrophic (PM), hetrotrophic (HT) and autotrophic (AT) conditions. Shoot length was the greatest (11.9cm) in 3$0^{\circ}C$ (HT) treatment and it was the shortest (3.4 cm) in $25^{\circ}C$ (PM) treatment. Nodal explants cultured in 3$0^{\circ}C$ treatment looked more vigorous than those of $25^{\circ}C$ in appearance, and node number was the greatest (10.5 per plantlet) among the treatments. But plantlets grew in 3$0^{\circ}C$ (HT) treatment were observed all overgrown. The size in leaf area was about 2 times greater and shoot length was about 2 times shorter in PM than in HT condition. Percent dry matter of shoots was 5.9% (HT) and 7.4% (PM) in $25^{\circ}C$ treatment and 6.1% (HT) and 7.4% (PM) in 3$0^{\circ}C$ treatment. Plantlets cultured in the MF+ treatments were less succulent than those cultured in the MF- treatment. Vitrified plantlets were examinated 14.8% (both $25^{\circ}C$ and 3$0^{\circ}C$) in PM condition and 22.2% ($25^{\circ}C$) and 31.5% (3$0^{\circ}C$) in HT condition. Sucrose was necessary for the survival of in vitro plantlets. In the sucrose-free medium, explants cultured in the MF- had turned yellow and were dead after 30 days of culture. But explants cultured in the MF+ were alive and produced plantlets with shoot and root (AT). On the other hand, the survival of explants on the MS basal medium (sucrose-free and hormone-free) depended entirely upon the MF attachment.

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Effects of Addition of Inorganic Germanium, GeO2 on the Growth, Germanium and Saponin Contents of Ginseng Adventitious Root in Submerged Culture (무기 게르마늄 GeO2의 첨가가 액체 배양 중 인삼 부정근의 생장과 게르마늄 및 사포닌 함량에 미치는 영향)

  • Chang, Eun-Jung;Oh, Hoon-Il
    • Journal of Ginseng Research
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    • v.29 no.3
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    • pp.145-151
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    • 2005
  • This study was carried out to determine the optimal submerged culture conditions for production of ginseng root containing germanium using plant tissue culture technology. The ginseng (Panx ginseng C.A. Meyer) .cot induced by plant growth regulators of 0.5 mg/L BAP and 3.0 mg/L NAA was cultured on SH medium and the effects of various $GeO_2$ concentrations, addition time of $GeO_2$ and pH of medium were investigated on fresh weight, saponin production and germanium accumulation in ginseng root. Optimal $GeO_2$ concentrations for fresh weight, saponin and germanium content were 10, 0 and 110ppm, respectively. When $GeO_2$ was added after 2 weeks cultivation of ginseng root, germanium content was higher than that of adding $GeO_2$ at the initial cultivation time, but saponin content and fresh weight were lower. pH 5.5 was found to be the most favorable condition for the growth of ginseng root and germanium accumulation, but saponin production was best at pH 6.0.

The Optimum Conditions for Induction of Ginseng Hairy Roots (인삼 모상근 유도를 위한 최적 조건)

  • 양덕춘;김용해;양덕조;신성련;최광태
    • Korean Journal of Plant Resources
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    • v.12 no.1
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    • pp.1-9
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    • 1999
  • The experiments were carried out to determine the optimum conditions for the induction of hairy roots in ginseng(Panax ginseng C.A. Meyer) by Agrobacterium spp. We were examined the antibiotics resistance of Agrobacterium spp and various ginseng parts, and the media for induction of hairy roots. The optimum concentration of NaOCl for sterilization of ginseng root segments without tissue damage with reduce of contamination was 7% NaOCl for 15-20 min and 9% NaOCl for 5 min, respectively. The more ginseng ages, the more contamination of ginseng root segment by sterilized in 7% NaOCl for 20 min, and especially in ginseng root segments with epidermis in six-year old roots. The growth of Agrobacterium spp were inhibited, but ginseng root segments was death in 30mg/L tetracycline. In 500mg/L cefotaxime or 500mg/L carbenicillin, the growth of Agrobacterium sup were inhibited, and root segments was grown normally. The optimum conditions for induction of hairy roots were using the root segments of three-year old ginseng cultured in 1/2MS medium supplemented with 500mg/L cefotaxime, and inoculation of Agrobacterium to root segments were better co-culture than smear method. After 2 weeks co-culture, the callus induced in cambium of root segments cultured in 1/2MS solid medium with 500mg/L cefotaxime. And then after 2 weeks, ginseng hairy roots were induced in callus of root segments. PCR analysis of rot C gene fragment confirmed that hairy roots were transgenic tissues.

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Comparison of Active Ingredients between Field Grown and In Vitro Cultured Rhizome of Korean Native Ginger (Zingiber officinale Roscoe) (조직배양생강과 한국재래종 생강의 유효성분 비교)

  • Jo, Man-Hyun;Ham, In-Ki;Lee, Gyu-Hee;Lee, Jong-Kug;Lee, Ga-Soon;Park, Sang-Kyu;Kim, Tae-Il;Lee, Eun-Mo
    • Korean Journal of Plant Resources
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    • v.24 no.4
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    • pp.404-412
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    • 2011
  • This study was carried out to compare and analyze the active ingredients of Korean native ginger and rhizome derived from in vitro shoot-tip culture of Korean native ginger. Proximate compositions, mineral nutrients, free sugars, fatty acids, volatile components, 6-gingerol, and 6-shogaol were analysed and evaluated. Korean native ginger was proved to have a little more contents than in vitro rhizome in proximate compositions (crude ash, crude lipid, crude protein, carbohydrate). Mineral nutrient contents (Cu, Fe, Mn, Zn) of in vitro rhizome were higher than those of Korean native ginger. Among the mineral nutrients, the quantity of K was the highest, followed by P, Mg, Na, and Ca. Free sugar contents (fructose, glucose, sucrose) of in vitro rhizome were higher than those of Korean native ginger. Fatty acids containing less than C14 was the major among the fatty acids in ginger. Citral ingredient of the unique aromatic compound of Korean native ginger was stronger than that of the rhizome derived from in vitro shoot-tip culture. Gingerol concentration was increased by shoot-tip culture.

Soil Acclimatization of Calanthe discolor through Multiple Shoot Formation from Tissue Culture (새우난초(Calanthe discolor)의 조직배양으로부터 다신초형성을 통한 토양순화)

  • Bae, Kee-Hwa;Yoon, Eui-Soo;Yun, Pil-Yong;Choi, Yong-Eui
    • Korean Journal of Plant Resources
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    • v.23 no.1
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    • pp.7-13
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    • 2010
  • This experiment was conducted to establish the micropropagation of Calanthe discolor through multiple shoot formation from the culture of leaf, corm and root explants. Frequency of adventitious shoot formation from leaf explants was higher than those of corms and root explants. Frequency of adventitious shoot formation on medium with various concentrations of BA (0. 1.0, 3.0, and 5.0 mg/L) and NAA (0, 0.1, 0.5, and 1.0 mg/L) was tested. The maximun induction of adventitious shoot was obtained on half strength Murashige and Skoog (MS) medium supplemented with 3.0 mg/L BA and 1.0 mg/L NAA after 6 weeks of culture. Multiple shoots were transferred onto half strength MS medium with various concentrations of GA3 (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L). The number and length of multiple shoots on medium were highest on medium with 3.0 mg/L GA3. All the adventitious shoot grew well and rooted on half strength MS medium with 3.0 mg/L NAA. The plantlets were acclimatized up to 100% on sand with TKS-II or pearlite with TKS-II.

Production of Gastrodia elata Tuber using Armillaria spp. (Armillaria 속균을 이용한 천마의 생산)

  • Sung, Jae-Mo;Jung, Bum-Shig;Yang, Keun-Joo;Lee, Hyun-Kyung;Harrington, T.C.
    • The Korean Journal of Mycology
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    • v.23 no.1 s.72
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    • pp.61-70
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    • 1995
  • The genus Armillaria is important because they produce Gastrodia tubers. Seventy two isolates of Armillaria were obtained from fruit bodies grown on decayed wood in Korea. Twenty four isolates from Pinus koraiensis were identified as A. ostoyae. Two isolates from G. elata growing in the field were identified as A. mellea. Seven isolates from Acer ginnala and Quercus spp. were identified as A. tabescens. Thirty nine isolates were identified as A. gallica. Armillaria gallica was isolated from Quercus spp., Ainus japonica, Vitis amurensis and Prunus sargentii. Armillaria spp. isolates were divided into four groups based on the cultural characteristics. Group II (A. gallica KNU-A110) was better than the other groups for mycelial growth and rhizomorph formation. Isolate KNU-A110 proved to be good for production of G. elata tubers. This fungus forms mycelial fan in the plant tissue and rhizomorphs in contact with G. elata tubers. Gastrodia spp. was found in thirteen sites in Kangweon province in Korea. The plants were divided into three different kinds based on stem color. Plants with stems of brownish orange and greyish yellow were identified as G. elata, and those with greyish green colored stems were identified as G. gracilis. Gastrodia was collected mainly from humus soils rich in leaf debris, and slopes facing south from mid-May to mid-July. Once the new tubers are formed from the ancestry tuber, the ancestry tuber begins to decay. The offspring tuber, apparently gaining nutrients through rhizomorphs, begins to grow in length and slowly to enlarge. It takes three years for the offspring tuber to become ancestry tuber.

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Growth and Quality Changes of Creeping Bentgrass by Application of Keratin Amino Acid Fertilizer (케라틴 아미노산 비료 시비에 따른 크리핑 벤트그래스의 생육과 품질 변화)

  • Jo, Gi-Woong;Kim, Young-Sun;Ham, Soun-Kyu;Lee, Jae-Pil;Kim, Doo-Hwan;Kim, Woo-Sung;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.5 no.4
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    • pp.260-267
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    • 2016
  • Amino acids in the plant were intermediate metabolites which produced by uptake and assimilation of nitrogen and these extracts which gained by bio-chemical digestion from protein of plant or animal were a source of functional fertilizer. This study was conducted to evaluate effects of keratin amino acid fertilizer (KAF) gained from animal hair or hoof on changes of turfgrass quality and growth by investigating turf color index, chlorophyll index, shoot number, clipping yield, and nutrient content in the turfgrass tissue. Treatments were designed as follows; non-fertilizer (NF), compound fertilizer (CF), keratin amino acid fertilizer treatments [CF + KAF $0.26ml\;m^{-2}$ (CKF), CF + KAF $0.52ml\;m^{-2}$ (2CKF)], and only keratin amino acid fertilizer treatment (KF). Shoot number, clipping yield, and nitrogen uptake of KF were higher than those of CF. As compared with CF, soil chemical properties, turf color index, chlorophyll index and clipping yield of keratin amino acid fertilizer were not significant, but shoot number and uptake of N and K were increased significantly. These results show that the application of keratin amino acid fertilizer increased shoot number and growth by increased uptake of nitrogen and potassium.

Scopolamine Production in Suspension Cultures of Tumor Calli from Datura metel L. (흰독말풀(Datura metel L.)종양 캘러스의 현탁배양으로부터 Scopolamine 생성)

  • 이수경;윤길영;김용해;양덕조
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.203-211
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    • 2000
  • In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

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