• Title/Summary/Keyword: Tissue Culture

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Difference of Microbiology according to Tissue Sampling in Diabetic Ulcers (만성 당뇨발에서 표재조직 및 심부조직 세균배양검사의 비교)

  • Rhee, Sung-Mi;Han, Seung-Kyu;Kim, Woo-Kyung
    • Archives of Plastic Surgery
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    • v.37 no.1
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    • pp.1-6
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    • 2010
  • Purpose: Diabetic foot infection is one of the most common and severe complications of diabetes mellitus that delays healing of the wound. Deep tissue biopsy is considered to be the gold standard method for antibiotic selection in treating infected chronic diabetic ulcers. However, swab culture or superficial tissue biopsy is often performed for a microbiologic test since deep tissue biopsy has limitations in application. The purpose of this study is to find out whether microbiologic results of swab culture or superficial tissue biopsy could be used for selection of antibiotics in treating diabetic ulcers. Methods: This study involved 42 patients with diabetic foot ulcers with negative results in bone probing test. Tissue samples for microbiologic tests were collected from all the patients by using superficial cotton swab, superficial tissue, and deep tissue. The microbiologic results of deep tissue biopsy were compared with swab culture and superficial tissue biopsy statistically. Results: Microbiology of the deep tissue showed the same results with those of the swab culture and superficial tissue in 67% and 71%, respectively. Statistical analysis demonstrated that the microbiology of the swab culture and superficial tissue does not coincide with that of the deep tissue. Conclusion: These results suggest that the microbiology of the swab culture and superficial tissue is not concordant with that of the deep tissue in infected chronic diabetic ulcers. To select appropriate antibiotic regimen, the speci specimen for the microbiologic test should be obtained from deep tissue.

Genetic Stability Studies in Micropropagated Date Palm (Phoenix dactylifera L.) Plants using Microsatellite Marker

  • Kumar, Nitish;Singh, Amritpal S.;Modi, Arpan R.;Patel, Armi R.;Gajera, Bhavesh B.;Subhash, Narayanan
    • Journal of Forest and Environmental Science
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    • v.26 no.1
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    • pp.31-36
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    • 2010
  • Sixteen microsatellite markers (simple sequence repeat (SSR) markers) were employed to examine the genetic stability of 27 randomly chosen date palm (Phoenix dactylifera L.) plants produced through somatic embryogenesis with upto forty two in vitro subcultures. No microsatellite DNA variation was observed among all micropropagated plants. Our results indicate that the micropropagation protocol used for rapid in vitro multiplication is appropriate and suitable for clonal propagation of date palm and corroborated that somatic embryogenesis can also be used as one of the safe modes for production of true-to-type plants of date palm. This is the first report on the use of microsatellite DNA markers to establish the genetic stability in micropropagated date palm plants.

Characteristics in Tissue Cultured Plants of Chinese foxglove(Rehmannia glutinosa) (지황 조직배양주의 수량성과 성분함량 특성)

  • 박충헌;성낙술;김선규;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.205-209
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    • 1999
  • Chinese foxglove(Rehmannia glutinosa) is receiving much attention as one of the principal medicinal crops and the crud drug. This study was conducted to obtain the basic breeding information of Chinese foxglove derived from tissue culture. To compare plant characteristics between local variety and tissue cultured seedlings, plant growth and root yield has been investigated. In addition, catalpol and free sugar contents were also analized. The ability for the storage of root stock originated from tissue culture seedlings were better than that of local variety. The growth and root yield of in vitro propagated plants were superior to those of conventionally propagated plants. Root yield of 1-year-old and 2-year-old seedlings investigated 112% and 246% respectively than Seacheon local shows 384 kg/10 a. Although there were no significant different between tissue culture seedlings and conventionally propagated one, slight decline of component contents in tissue culture plants were still existed. Investigated sugar content among Seocheon local, 1-year-old tissue culture seedlings, and 2-year-old tissue culture seedlings were 1.86%, 1.21%, and 1.10% respectively. Catalpol content as one of standard materials indicates 0.46% in Seocheon local and 0.40% in tissue cultured ones.

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An Effective Detection of Potato Virus Y Using RT-PCR Technique (RT-PCR 기법을 이용한 효과적인 감자바이러스 Y의 검정)

  • Joung, Young-Hee;Jeon, Jae-Heung;Choi, Kyung-Hwa;Kim, Hyun-Soon;Yi, Yong-Sub;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.219-224
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    • 1997
  • A PT-PCR (reverse transcription-polymerase chain reaction) diagnostic method for potato virus Y (PVY) was developed using primer pair derived from conserved region of coat protein genes of several PVY strains, A 764 bp PCR product was detected from several lines of potato cv. Atlantic. We could prove that the 764 bp DNA fragment was indeed the PVY gene by sequencing analysis. PVY detection method using RT-PCR technique was about tuber tissue.

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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Production of Azadirachtin from Plant Tissue Culture: State of the Art and Future Prospects

  • Prakash, Gunjan;Bhojwani, Sant S.;Srivastava, Ashok K.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.4
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    • pp.185-193
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    • 2002
  • With Increasing awareness towards environment-friendly and non-toxic pesticide azadirachtin obtained from neon tree (Azadirachta indica) is gaining more and more importance. Its broad-spectrum activity, Peculiar mode of action. eco-friendly and non-toxic action towards beneficial organisms has offered many advantages over chemical pesticides. All currently use commercial formulations based on azadirachtin contains azadirachtin extracted from seeds of naturally grown whole plants which is labour intensive process depending upon many uncontrollable geographical and climatic factors. Plant tissue culture can be a potential process for the pro-duction, offering consistent, stable and controlled supply of this bioactive compound, However the research on tissue culture aspects of production are in preliminary stage and requires culture and process optimization for the development of a commercially viable process. This review states the present status and future challenges of plant tissue culture for azadirachtin production.

Expression of Chitinase Gene in Solanum tuberosum L.

  • Park, Kyung-Hwa;Yang, Deok-Chun;Jeon, Jae-Heung;Kim, Hyun-Soon;Joung, Young-Hee;Hyouk Joung
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.85-90
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    • 1999
  • In order to protect fungal diseases, leaf disc explants of Solanum tuberosum cultivar, Belchip, was infected with an Agrobacterium MP90 strain containing chimeric gene construct, consisting of antibiotic resistance and chitinase gene driven by the CaMV 35S promoter, for transformation. Regenerated multiple shoots were selected on a medium containing kanamycin and carbenicillin after exposure to Agrobacterium. The presence and integration of the npt II and chitinase gene were confirmed by polymerase chain reaction(PCR). Northern blot analysis indicated that the genes coding for the enzyme could be expressed in potato plants. The chitinase activity of transgenic potato plants was higher than the control potato.

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Detection of Lily Symptomless Virus Using RT-PCR Technique (RT-PCR 기법을 이용한 Lily Symptomless Virus의 검정)

  • Joung, Young-Hee;Jeon, Jae-Heung;Choi, Kyung-Hwa;Kim, Hyun-Soon;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.187-190
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    • 1996
  • 백합으로부터 total RNA를 분리하여 LSV 외피단백질 유전자의 551 bp에 해당하는 특정 염기서열을 증폭할 수 있는 primer로 RT-PCR를 수행하였다. 그 결과 Lilium oriental hybrid cvs. Miani, Marco Polo, Casablanca, Le Reve 품종에서 551 bp의 DNA 절편이 증폭되었고 이 절편의 염기서열을 분석한 결과 LSV외피단백질 유전자의 일부임을 확인할 수 있었다. 그러므로 RT-PCR 방법으로, 실험에 사용하 s4품종 모두 LSV에 감염되어 있음을 알 수 있었다.

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Aortic and Pulmograft Transplantation Utilizing Cryopreservation (초저온 냉동보관법을 이용한 동종판막 이식술에 대한 연구)

  • Song, Myeong-Geun;Lee, Dong-Sun
    • Journal of Chest Surgery
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    • v.23 no.4
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    • pp.622-639
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    • 1990
  • The use of aortic valve homograft has been developed since 1962 when Ross and Barratt - Boyes independently replaced a diseased aortic valve with an orthotopically inserted homograft valve. And also surgical treatment of complex congenital cardiac malformations utilizing homograft extracardiac conduit has been tried with better result than any other prosthetic material. The present study was undertaken to clarify the safety tissue viability, sterility, after following our protocol of procurement of heart, dissection of aortic and pulmonic homograft, sterilization, cryopreservation, thawing and dilution, and transplantation on experimental animal, sheep. Tissue viability of valve and great artery was assessed by tissue culture. Sterility was evaluated by bacterial and fungal culture. The method used was proven no deleterious effect on the integrity of the valve. Tissue culture of valve tissue before, and after cryopreservation process resulted that active fibroblast growth was observed from homograft sterilized with antibiotics. And culture of the transplanted homograft from sacrificed animal showed active fibroblast growth. Pathologic examination of implanted valve tissue from sacrificed sheep showed mild calcification and minor change, but there were moderate and severe calcification of wall of great arteries.

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Effect of Cytokinins on the Proliferation of Multiple Shoots in Horsegrgm [Macrotyloma uniflorum (Lam.) 'Verdc.]

  • Mohamed, Shamsudeen Varisai;Jawahar, Manikam;Thiruvengadam, Muthu;Jeyakumar, Masilamani;Jayabalan, Narayanasamy Pillai
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.79-83
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    • 1999
  • A method for induction of multiple shoots using cotyledonary nodes and shoot tips of Macrotyloma uniflorum (Lam.) Verdc. was described. The experiment was conducted in which shoot induction was noticed on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of four cytokinins (KIN, 2iP, Ads, BAP). These multiple shoots were later developed into normal shoots. The highest rate of shoot proliferation came from MS medium added with BAP 1.5 mg/L. The multiple shoot buds were subcultured into MS medium with BAP (0.5-1.5 mg/L) along with Ads (1.0 mg/L) and GA$_3$ (0.5 mg/L), which gave rise to the highest frequency of shoot proliferation and elongation. The shoots were rooted on MS medium supplemented with 1.75 mg/L IBA.

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