• Title/Summary/Keyword: in vitro shoots

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Growth-Inhibiting Effects of Various Traditional Drinks of Plant Origin on Human Intestinal Bacteria

  • Jeon, Ho-Joung;Roh, Jung-Yeon;Kim, Do-Hyoung;Lee, Wan-Kyu;Ahn, Young-Joon
    • Applied Biological Chemistry
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    • v.41 no.8
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    • pp.605-607
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    • 1998
  • A total of 28 traditional drinks derived from 23 plant species in 19 families were tested for their in vitro growth-inhibiting effects against Bifidobacterium adolescents, Bifidobacterium bifidum, Lactobacillus acidophilus, Lactobacillus casei, Clostridium perfringens, and Escherichia coil using a paper disc agar diffusion method under anaerobic condition. The responses varied with bacterial strain, plant species and tissue sampled. In a test with C. perfringens at 5 and 10 mg/disc, potent growth inhibition was produced from the extracts of Eucommia ulmoides stems, Pinus densiflora leaves and shoots, Thea sinensis leaves (green and oolong teas) and Zingiber officinale roots. All materials tested did not adversely affect the growth of bifidobacteria, lactobacilli, and E. coli. These results may be an indication of at least one of the pharmacological activities of these plant-derived drinks.

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High Frequency Regeneration of Plantlets from Seedling Explants of Asteracantha longifolia (L.) NEES

  • Mishra Ramya Ranjan;Behera Motilal;Kumar Deep Ratan;Panigrahi Jogeswar
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.27-35
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    • 2006
  • Plantlet regeneration in Asteracantha longifolia(L.) Nees (Acanthaceae), a medicinal herb has been achieved from seedling explants on basal MS medium. Three different seedling explants including node, internode and leaf segments on used. Of these three explant, leaf explants gave better response for both callus mediated organogenesis and direct multiple shoot induction. Number of explants showing differentiation of shout buds was higher on MS media supplemented with BA compared to kinetin. MS medium fortified with BA ($2.0mgl^{-1}$) and NAA ($0.5mgl^{-1}$) was found to be most suitable for both callus mediated organogenesis and elongation of shouts. The elongated shoots were successfully routed on MS medium fortified with NAA or IBA. Among them $0.1mgl^{-1}$ NAA or $0.2mgl^{-1}$ IBA provides better response for rhizogenesis. Regenerated plantlets were successfully established in soil where 85.4% or them developed into morphologically normal and fertile plants. RAPD profiling using four decamer primers confirmed the genetic uniformity of the regenerated plantlets and substantiated the efficacy and suitability of this protocol for in vitro propagation of A. longifolia.

Expression of Organogenesis-related Genes and Analysis of Genetic Stability by ISSR Markers of Regenerants Derived from the Process of in vitro Organogenesis in Japanese Blood Grass (Imperata cylindrica 'Rubra') (기내배양 홍띠 단계별 재분화체의 기관분화 관련 유전자 발현과 ISSR에 기반한 유전적 안정성 분석)

  • Ye-Jin Lee;In-Jin Kang;Chang-Hyu Bae
    • Korean Journal of Plant Resources
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    • v.36 no.5
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    • pp.496-507
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    • 2023
  • The in vitro organogenesis is one of important issues in plant embryology, and somaclonal variations are existing in calli and/or regenerants induced from a process of the organogenesis with in vitro circumstances. In this study, expressions of organogenesis-related genes were evaluated and genetic stability of regenerants derived from the process of in vitro organogenesis were measured using ISSR markers in Imperata cylindrica 'Rubra', Poaceae. The expressions of organogenesis-related genes were detected all of regenerants at the process of the organogenesis. All ISSR markers produced with an average of 71 bands per in vitro-cultured regenerants, and the scorable bands were varied from two to eight with an average of 5.14 bands per a primer. The polymorphism rates of the in vitro regenerants were higher than that of mother plants (1.4%), showing 4.1% (pot-cultured regenerants), 4.3% (field-cultured regenerants), 4.2% (in vitro-cultured regenerants), 5.6% (calli with green shoots) and 1.4% (calli), respectively. The genetic similarity matrix (GSM) among all accessions ranged from 0.747 to 1.0 with a mean of 0.868. GSM of the regenerants showed differences (from 0.972 to 1.00) compared with that of mother plants (0.991). According to the clustering analysis, two independent groups were divided into; the one is mother plants and regenerants cultured at room and open field, the other is regenerants cultured in vitro. The results give a new insight for understanding the dynamics of organogenesis in monocot plant.

Herbicide Resistant Cabbage (Brassica oleracea ssp. capitata) Plants by Agrobacterium-mediated Transformation

  • Lee, Yeon-Hee;Lee, Seung-Bum;Suh, Suk-Chul;Byun, Myung-Ok;Kim, Ho-Il
    • Journal of Plant Biotechnology
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    • v.2 no.1
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    • pp.35-41
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    • 2000
  • Transgenic cabbage (Brassica oleracea ssp. capitata) plants resistant to the commercial herbicide Bast $a^{R}$ were obtained by Agrobacterium tumefaciens - mediated transformation. Hypocotyl segments of in vitro grown plants were infected with Agrobacterium tumefaciens LBA 4404 harboring plasmid pMOG6-Bar which contains hpt and bar genes. Explants were cultured on callus induction medium (MS basal medium + 1 mg/L NAA + 2 mg/L BA + 2 mg/L AgN $O_3$+ 100 mg/L carbenicillin + 250 mg/L cefotaxime) supplemented with 15 mg/L hygromycin. Hygromycin resistant calluses were transferred to shoot regeneration medium (MS basal medium + 0.1 mg/L NAA + 2 mg/L BA + 3% sucrose + 2 mg/L AgN $O_3$+ 15 mg/L hygromycin + 250 mg/L cefotaxime + 100 mg/L carbenicillin). In order to induce roots, elongated shoots were placed on the MS medium without plant growth regulators and hygromycin. Southern blot analysis of several putative transgenic plants indicated that one to five intact copies of Apt and bar genes were incorporated into the genome. Expression of bar gene was confirmed by Northern blot analysis and by herbicide resistant phenotype. Seed progeny from self-pollinated transformants expressed the herbicide resistance and showed Mendelian segregation of the introduced gene.e.

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Effect of nitrogen sources and 2, 4-D treatment on indirect regeneration of ginger (Zingiber officinale Rosc.) using leaf base explants

  • Mehaboob, Valiyaparambath Musfir;Faizal, Kunnampalli;Raja, Palusamy;Thiagu, Ganesan;Aslam, Abubakker;Shajahan, Appakan
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.17-21
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    • 2019
  • Ginger is an important monocotyledonous plant belonging to the family Zingiberaceae. The objective of this study was to investigate the regeneration potential of ginger using leaf base explants. Auxins such as 2, 4-D and NAA in combination with BA were used for initiation of callus. Different combinations of both ammonium ($NH^{4+}$) and nitrate ($NO^{3-}$) were also studied for efficient callus production. High frequency of white friable calli was observed on modified Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 2, 4-D, 0.5 mg/L NAA and 0.5 mg/L BA. The highest shoot induction (92.33%), shootlets number ($7.33{\pm}0.33$) and length ($88.33{\pm}4.40$) mm were achieved on MS media containing 0.5 mg/L BA. Regenerated shoots were transferred to in vitro rooting media containing 1.0 mg/L IBA. Afterwards, plantlets with well-developed root and shoot system were subjected to a twostep hardening process. 71% of plantlets survived after secondary hardening without any abnormal morphology.

Efficient and Reliable in vitro Regeneration System for Rubus Species as the Basis of Genetic Engineering

  • Kalai Katalin;Meszaros Annamaria;Denes Ferenc;Zatyko Jozsef;Balazs Ervin
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.241-246
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    • 2005
  • Factors affecting regeneration of different Rubus varieties (blackberry, raspberry and their hybrid) were examined and a reliable regeneration system was established. Media for stock plant maintenance were tested; different explants and media were investigated to find the best circumstances for the regeneration. The effect of the commonly used antibiotics was studied to determine the most suitable one for selection of the transformants. We found that both MS and LS media supplemented by $20\;gL^{-1}$ sucrose are suitable for the stock plant maintenance. The optimal hormone content for the stock plants is $0.125\;mgL^{-1}$ 6-benzylaminopurine (BAP) with $0.01\;mgL^{-1}$ indole-3- butyric acid (IBA). The highest regeneration rate was observed on medium containing MS salts with B5 vitamins complemented with glucose, sucrose, maltose, $10\;gL^{-1}$ each, supplemented with benzylaminopurine riboside (BAR) ($2\;mgL^{-1}$) and indole-3-acetic acid (IAA) ($0.1\;mgL^{-1}$). The regenerated shoots appeared directly from the cut edges, without callus phase. Hygromycin and geneticin proved to be good selection agents for the Rubus explants, but due to their severe effect on the tissues we propose to use marker-free constructions for the transformation.

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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In Vitro Screening of Antibacterial Agents for Suppression of Fire Blight Disease in Korea (기내 검정법을 이용한 국내 과수 화상병 방제제 선발)

  • Lee, Min Su;Lee, Ingyeong;Kim, Sam Kyu;Oh, Chang-Sik;Park, Duck Hwan
    • Research in Plant Disease
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    • v.24 no.1
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    • pp.41-51
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    • 2018
  • Since fire blight disease on apple and pear was produced in Korea in 2015, there were no registered chemicals to control against this disease. Instead, several antibacterial chemicals that were registered for other bacterial diseases such as soft rot and bacterial spot have been authorized by Rural Development Administration (RDA). However, these chemicals are not tested efficacy for fire blight disease except damage by those treatments on apple and pear in Korea. Thus, we evaluated efficiency using in vitro and in planta assays of antibacterial chemicals such as antibiotics and copper compounds including kasugamycin, oxytetracycline, oxolinic acid and streptomycin, and copper hydroxide, copper sulfate, oxine copper and tribasic copper sulfate, respectively. We also tested two kinds of biological agents. As expected, significant antibacterial effect was observed in vitro test of both antibiotics and copper-based chemicals. In planta test based on disease severity including ooze and water-soaked formation on immature pears, bacterial populations on blooms, and blight lesion formation in artificially inoculated shoots, kasugamycin, oxytetracycline and streptomycin have been shown the most efficiency among tested antibiotics. Four copper-based chemicals tested in this study, control effects are little bit lower than agricultural antibiotics but they seem to be available to use in terms of winter season. Biocontrol agents were also shown possibility to treat in eco-friendly farms. In addition, there are no antibiotic resistance genes in Korean isolates against antibiotics, which were selected for suppression of fire blight in this study.

Strategy for Bio-Diversity and Genetic Conservation of Forest Resources in Korea (생물종(生物種) 다양성(多樣性) 및 삼림유전자원(森林遺傳資源) 보존(保存) 전략(戰略))

  • Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.83 no.2
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    • pp.191-204
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    • 1994
  • Due to its topographic complexities and various climatical condition, Korea exhibits diverse forest types. Dominant tree species in this zone are Quercus spp., Betula spp., Zelkova spp., Fraxinus spp., Pinus densiflora, Pinus koraiensis, and Pinus thunbergii ete. Genetic conservation in forest species in Korea there are three ways ; one is in situ, other is ex situ and third is in-facility conservation. In situ conservation include that are the present status of conservation of rare and endangered flora and ecosystem, the reserved forest, the national and provincial park, and the gene pool of natural forests. Ex situ conservation means to be established the new forest from in situ forest stands, progeny and provenance test populations, seed orchard and clone banks, and gene conservation in-facility. As a tool for low temperature storage, several aspects on in vitro system were studied ; (1) establishment of in vitro cultures from juvenile and/or rejuvenated tissues, (2) induction of multiple shoots from the individual micropropagules, (3) elongation of the proliferated shoots. Studies on cold storage for short-and long-term maintenance of in vitro cultures under $4^{\circ}C$ in the refrigerator were conducted. For the cryopreservation at $-196^{\circ}C$, various factors affecting survivability of the plant materials are being examined. The necessity of gene conservation of forest trees is enlarged not only to increase the adaptability for various environments but also to gain the breeding materials in the future. For effective gene conservation of forest trees, I would like to suggest followings ; 1. Forest stands reserved for other than the gene conservation purposes such as national parks should be investigated by botanical and gene-ecological studies for selecting bio-diversity and gene conservation stands. 2. Reserved forest for gene pool should be extented both economically important tree spp. and non-economical species. 3. Reserved forest for progeny test and clone bank should be systematically investigated for the use of Ex situ forest gene conservation. 4. We have to find out a new methodology of genetic analysis determining the proper and effective size of subpopulation for in situ gene conservation. 5. We should develop a new tree breeding systems for successful gene conservation and utilization of the genetic resources. 6. New method of in-facility gene conservation using advanced genetic engineering should be developed to save time and economic resources. 7. For the conservation of species with short-life span of seed or shortage of knowledge of seed physiology, tissue culture techniques will be played a great role for gene conservation of those species. 8. It is are very useful conservation not only of genes but of genotypes which were selected already by breeding program. 9. Institutional and administrative arrangements including legistlation must be necessarily taken for gene conservation of forest trees. 10. It is national problems for conservation of forest resources which have been rapidly destroyed because of degenerating environmental condition and of inexperienced management system of bio-diversity and gene conservation. 11. In order to international cooperation for exchanging data of bio-diversity and gene conservation, we should connect to international net works as soon as possible.

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Propagation of Acorus gramineus from Seeds and In vitro Culture (종자 및 기내배양을 이용한 석창포 증식)

  • Park, Young-Chul;Kim, Jeong-Seon;Yang, Seok-Chul;Cho, Youn-Dong;Kim, Yong-Duk;Park, Jae-Kweon
    • Korean Journal of Plant Resources
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    • v.21 no.5
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    • pp.347-351
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    • 2008
  • Rhizomes of Acorus gramineus Soland have been used as sedatives, analgesics, and stomachics in Korean medicine. Even though A. gramineus produced in Korea is known as having better efficacy of a remedy than that in China, its cultivation area has not increased because of the lack of seedlings. To solve this problem, seed propagation method was tested. Seeds were harvested and sowed three times in June, 2005. The best results showed at greenish yellow seed stage harvested in mid June, and its germination ratio was 90.7%. Seeds were well stored at $5^{\circ}C$ for 30 days. As for in vitro culture, multiple shoots were induced first in MS basal medium supplemented with $2.0mg{\cdot}L^{-1}$ NAA plus $0.1mg{\cdot}L^{-1}$ BA, and then roots were induced in MS basal medium containing $0.1mg{\cdot}L^{-1}$ BA or $0.5mg{\cdot}L^{-1}$ NAA plus $1.0mg{\cdot}L^{-1}$ BA. Growth of A. gramineus seedlings in Wonyesangto, potting material sold in markets for horticultural plants, was superior to vermiculite.