• Title/Summary/Keyword: root tissue

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Overexpression of Hyoscyamine 6${\beta}$-Hydroxylase (h6h) Gene and Enhanced Production of Tropane Alkaloids in Scopolia parviflora Hairy Root Lines

  • KANG, YOUNG-MIN;LEE, OK-SUN;JUNG, HEE-YOUNG;KANG, SEUNG-MI;LEE, BYUNG-HYUN;CHANDRAKANT KARIGAR;THEERTHA PRASAD;BAHK, JUNG-DONG;CHOI, MYUNG-SUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.91-98
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    • 2005
  • The hyoscyamine 6${\beta}$-hydroxylase (h6h) gene was introduced into the genome of Scopolia parviflora through the Agrobacterium rhizogenes binary vector system. The enzyme was expressed ally and tissue specific selectively in roots, resulting in five transgenic hairy root lines. The presence of the h6h gene in kanamycin-resistant hairy roots and its overexpression were confirmed by polymerase chain reaction (PCR), Northern blotting, and Western blotting, respectively. In the transgenic hairy root lines which constitutively expressed the H6H enzyme, hyoscyamine and scopolamine accumulated in high concentration. Among the transgenic hairy root lines that expressed the H6H enzyme, only two were more productive. The levels of tropane alkaloids in transgenic hairy root varied greatly: The best transgenic line (#5) contained 8.12 mg of scopolamine per g dry weight, which produced the compound three times more than wild-type root. These results suggest a possibility of improving the yield of tropane alkaloids in hairy root lines by genetic and metabolic engineering.

Effect of Explant Types, Auxin Concentration and Light Condition on In Vitro Root Production and Alkaloid Content of Rauvolfia serpentina (L.) Benth. ex Kurz

  • Yahya, Andi Fadly;Hyun, Jung-Oh;Lee, Jae-Ho;Jung, Myung-Suk
    • Journal of Korean Society of Forest Science
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    • v.96 no.2
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    • pp.178-182
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    • 2007
  • Rauvolfia serpentina (L.) Benth. ex Kurz is a medicinal plant and an endangered tropical rainforest plant species. Since the field cultivation that aims to fulfill the industrial needs is never accomplished, tissue culture appears to be the most feasible way to improve the quality and quantity of R. serpentina. This experiment used two kinds of explants (roots and shoots) to induce optimal root formation in different combinations of auxin and photoperiod. Each explants exhibited different responses on given treatments. Differentiated root could be produced from explants cultured in IBA 20 mg/L with and without light. The highest number of roots, root length and root weight induced from shoot explants were effective on MS medium containing IBA 20 mg/L and incubated under dark condition, while highest total weight (callus and root) from root explants cultured on MS medium supplemented 10 mg/L IBA and 10 mg/L NAA and incubated under day length (11/13 hr). The root induced from shoot explants produced the highest major alkaloid content. The highest content of ajmaline (2.17 ppm fresh weight) and reserpine (1.30 ppm fresh weight) were observed in shoot explants cultured in MS medium containing combination of IBA 10 mg/L and NAA 10 mg/L and incubated under dark condition, yohimbine (1.47 ppm fresh weight) was in the shoot explants cultured in MS medium containing NAA 20 mg/L and incubated under day length, while serpentine was absent.

New Antisense RNA Systems Targeted Against Plant Pathogens

  • Matousek, J.;Vrba, L.;Kuchar, M.;Pavingerova, D.;Orctova, L.;Ptacek, J.;Schubert, J.;Steger, G.;Beier, H.;Riesner, D.
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.379-385
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    • 2000
  • tRNA and 7SL RNA based antisense vehicles were prepared by inserting conserved anti-viral and anti-viroid domains. Anti-PVS coat protein leader sequence (ACPL) and antistructural antihairpin domain of PSTVd (AHII) were inserted in tRNA cassette; anti- zing finger domain of PVS, AHII and anti hop latent viroid ribozyme were inserted in 7SL RNA gene isolated from A. thaliana. These constructs were shown to be transcribed both, in in vitro and in in vivo conditions. However, it followed from our work that closely linked position of PoIII reference genes and PoIIII antisense genes within T-DNA lead to the impairment of RNA expression in transgenic plants. To assay in vivo transcription of antisense genes, hairy root potato cultures were established using h. tumefaciens A4-24 bearing both, Ri plasmid and PoIII-promoterless plant expression vectors with antisense RNA genes. Expression of antisense RNA in transgenic potato tissues was proven by specific RT-PCR reactions.

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Effect of Nutrient Concentrations and Leaching Percentage on Growth and Nutrient Uptake by Perilla Frutesens Britton var. Japonica Hara in Plug Culture (잎들깨의 플러그육모에서 용탈률 및 시비농도가 생육 및 무기원소 흡수에 미치는 영향)

  • Choi, Jong-Myung;Yoon, Hwa-Mo;Park, Jong-Yoon
    • The Journal of Natural Sciences
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    • v.13 no.1
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    • pp.83-96
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    • 2003
  • Effect of Nutrient Concentrations, fertigation frequency, and learching percentage on crop growth and nutrient concentrations in root media were evaluated. The treatment of each irrigation with $50 mg.L^{-1}$ of nitrogen in stage 2 and increase to $80 mg.L^{-1}$ nitrogen in stage 3 had the highest crop growth at 34 days after sowing among treatments tested. Feeding with low nutrient concentrations and elevated frequency decreased crop growth. In treatments of each leaching percentage, feeding with low nutrient concentrations and elevated frequency resulted in increased tissue nutrient contents. The less tissue potassium content and higher calcium and magnesium contents were observed in treatment of 50% leach than those in 0% leach. All treatment tested had soil solution pH higher than 6.8. Electrical conductivity in treatments of 50% leach were lower than those of 0% leach. Feeding with low nutrient concentrations and elevated feeding frequency in each leaching percentage resulted in increased electrical conductivity in soil solution of root media. Trends of medium nutrient concentrations were similar to those of electrical conductivity.

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'A COMPARATIVE HISTOLOGIC STUDY OF BONE-IMPLANT INTERFACE TO THE TITANTIUM ROOT FORMED IMPLANTS IN THE Mx, Mn' (상악 및 하악골에 식립된 치근형 골내매식체와 주위골조직의 반응에 관한 조직학적 비교연구)

  • Lee, Jae-Hwang;Hur, Song-Ju;Cho, In-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.29 no.2
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    • pp.35-48
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    • 1991
  • Installation periods of implants in Mx. and Mn., is related to pattern of bone formation. The purpose of this study was to observe histologic response in osseointegration at root formed implant-tissue interface at Mx. and Mn., the other is comparison of osseointegration level between Mx. and Mn. at 8 weeks. In this study, unilateral upper & lower molars were extracted in dog. After allowing to heal for 4 months, two kinds of osseointegrated implants Swedevents, Corevents-were inserted in dog. The specimens were treated by conventional method. The interface zones between bone and implant were investiigated using X-rays, light microscope. The following results were obtained from this study. 1. Around titanium implants that were installed in Ma and Mn., Radio lucencies don't exist 2. There are not inflammation and mobility of titanium implants that were installed in Mx. and Mn. Most of implant surface are covered by bony tissued partly by bone-marrow tissues. 3. Titanium implants installed in Mx, in contrast to same implants in Mn., shows more coverage by bone marrow tissue and lack of apposition lamellar bone, which lead to the assumption that bone formation in Mn. is faster than in Mx.

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Differential Responses of Rice Acid Phosphatase Activities and Isoforms to Phosphorus Deprivation

  • Lim, Jeong-Hyun;Chung, Ill-Min;Ryu, Sang-Soo;Park, Myoung-Ryoul;Yun, Song-Joong
    • BMB Reports
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    • v.36 no.6
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    • pp.597-602
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    • 2003
  • Acid phosphatases (APases) play a role in the release of phosphate in organic complexes in soil. We investigated tissue- and isoform-specific responses of APases to phosphorus (P) deficiency in three rice genotypes; Dasan-byeo, Sobi-byeo, and Palawan. The levels of shoot APase activity per protein were similar in the three genotypes. They significantly decreased with P deprivation that was longer than seven days. Root APase activity per protein was two- to three-fold higher in Dasan than in Sobi and Palawan. In all genotypes the APase activity increased in P-deficient plants, but the increase was higher in Sobi and Palawan. After 21 days of P deprivation, secreted APase activity increased more than eight-fold in Dasan and two-fold in Sobi and Palawan. Isoform profiles of shoot and root APases were most diverse in Dasan. The activities of the major isoforms in P-deficient shoots decreased in all three genotypes. Depending on the genotypes, further increases in constitutive isoforms and new induction of one to four isoforms occurred in P-deficient roots. The results indicate that tissue and genotype differences in the response of APase to P deficiency are primarily facilitated by the different responses of the isoforms.

Apicoectomy of maxillary anterior teeth through a piezoelectric bony-window osteotomy: two case reports introducing a new technique to preserve cortical bone

  • Hirsch, Viola;Kohli, Meetu R.;Kim, Syngcuk
    • Restorative Dentistry and Endodontics
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    • v.41 no.4
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    • pp.310-315
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    • 2016
  • Two case reports describing a new technique of creating a repositionable piezoelectric bony window osteotomy during apicoectomy in order to preserve bone and act as an autologous graft for the surgical site are described. Endodontic microsurgery of anterior teeth with an intact cortical plate and large periapical lesion generally involves removal of a significant amount of healthy bone in order to enucleate the diseased tissue and manage root ends. In the reported cases, apicoectomy was performed on the lateral incisors of two patients. A piezoelectric device was used to create and elevate a bony window at the surgical site, instead of drilling and destroying bone while making an osteotomy with conventional burs. Routine microsurgical procedures - lesion enucleation, root-end resection, and filling - were carried out through this window preparation. The bony window was repositioned to the original site and the soft tissue sutured. The cases were re-evaluated clinically and radiographically after a period of 12 - 24 months. At follow-up, radiographic healing was observed. No additional grafting material was needed despite the extent of the lesions. The indication for this procedure is when teeth present with an intact or near-intact buccal cortical plate and a large apical lesion to preserve the bone and use it as an autologous graft.

A SCANNING ELECTRON MICROSCOPIC STUDY OF THE CLEANSING EFFECT OF RC-PREP ON THE DENTINAL WALLS OF THE ROOT CANAL (RC-Prep의 근관정화효과(根管淨化效果)에 관(關)한 주사전자현미경적(走査電子顯微鏡的) 연구(硏究))

  • Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.7 no.1
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    • pp.65-69
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    • 1981
  • This study was conducted to evaluate the cleansing effect of RC-Prep (10% Urea-peroxide, 15% EDTA) on apical third of root canal. Thirty single rooted human teeth were divided into three groups, and ten canals in each group were all enlarged three sizes greater than their original diameter with K-type files and irrigated with each of three irrigants. The three used irrigants were RC-prep (Premier Co.) in combination with 3.5% Sodium hypochlorite, Normal saline, 3.5% Na OCL. In each group, one of three irrigants were used in conjunction with instrumentation as they would be during clinical condition. After final irrigation, the canals were dried with paper points and the teeth were split longitudinally. The cleanness of canal walls iii the apical region were examined with Scanning Electron Microscope. The following results were drawn. 1. The use of Rc-Prep in combination with 3.5% NaOCl showed more clean canal surface than the use of other two irrigants. 2. The canals used RC-Prep in combination with 3.5% NaOCl revealed remnants of pulp tissue and smeared layer, but the openings of dentinal tubules were relatively clean and wide. 3. There was no significant difference in the debridement effect of 3.5% NaOCl and Saline solution. 4. The use of Saline solution showed great amount of remnants of pulp tissue which couldn't find in the use of 3.5% NaOCl.

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Diels-Alder Type Adducts from Hairy Root Cultures of Morus macroura

  • Happyana, Nizar;Hakim, Euis H.;Syah, Yana M.;Kayser, Oliver;Juliawaty, Lia D.;Mujahidin, Didin;Ermayanti, Tri M.;Achmad, Sjamsul A.
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.233-237
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    • 2019
  • Three Diels-Alder type adducts, guangsangon E (1), chalcomoracin (2) and sorocein I (3) were isolated from hairy root cultures of Morus macroura. The structures of the isolated compounds (1-3) were determined by spectroscopic method (NMR and MS), and spectral comparison to literature. Cytotoxic activities of the isolated compounds (1 - 3) were investigated against P-388 murine leukemia cell line. Guangsangon E (1) showed the most potent cytotoxicity against P-388 murine leukemia cell line with $IC_{50}$ value of $2.75{\pm}0.32{\mu}g/mL$. To the best of our knowledge, guangsangon E (1) and sorocein I (3) were reported for the first time from the tissue cultures of M. macroura.

Effects of plant preservative mixtureTM on in vitro germination of Dendrobium thyrsiflorum Rchb.f. and its application in orchid conservation

  • Tran Trung Chanh;Nguyen Tan Huy;Nguyen Thu Ha;Khanh Le;Nguyen Huu Hoang
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.108-114
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    • 2023
  • In vitro conservation is one of the most effective strategies for rare plant protection, especially for orchid species. To maximize the success rates of in vitro explant establishment (stage I) in conservation programs, the application of tissue culture additives such as Plant Preservative MixtureTM (PPMTM) should be emphasized. In this study, we used Dendrobium thyrsiflorum Rchb.f. (1875) seeds and seedlings as a model for the evaluation of PPMTM's phytotoxicity in the meristematic tissues of epiphytic orchids. PPMTM had no observable inhibitory effect on protocorm, shoot, or root development when it was supplemented at 0.1%. PPMTM supplementation caused adverse effects on D. thyrsiflorum explants at concentrations > 0.2%. At high concentrations, young in vitro seedlings showed damage, especially at the root tissue level. Based on this model, supplementation of 0.1-0.2% PPMTM to culture media was successfully implemented to establish in vitro cultures of other rare orchid species in our conservation program.