• Title/Summary/Keyword: in vitro conservation

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Conservation of An Endangered Species of Lilium cernum Komarvo. through in vitro Mass-Propagation (멸종위기 종 솔나리의 기내 대량생산을 통한 보존 연구)

  • Lee, Song-Hee;Lee, Su-Gwang;Kang, Ho-Duck
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.1
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    • pp.9-15
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    • 2011
  • This study was conducted to establish the optimal condition for conservation of genetic resources and micropropagation of Lilium cernum. Induction of bulbet of L. cernum was highly effective (9.2 bulb/explant) on 1/2 SH (Schenk and Hildebrandt) medium supplemented with 1.0 mg/L TDZ (Thidiazuron) and 0.1 mg/L NAA (Naphthaleneacetic acid). The treatment of 0.1 mg/L NAA increased root development (6.4 root/explant) under the in vitro condition. In addition, treatments of AC (Activated Charcoal) and ventilation were enhanced to develop number of shoots and to elongate length of leaf, bulb and root. Futhermore, the process of short-term soil acclimatization was promoted to strengthen the plantlets induced under the in vitro condition.

Identification of the Genes Involved in the Fruiting Body Production and Cordycepin Formation of Cordyceps militaris Fungus

  • Zheng, Zhuang-Li;Qiu, Xue-Hong;Han, Ri-Chou
    • Mycobiology
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    • v.43 no.1
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    • pp.37-42
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    • 2015
  • A mutant library of Cordyceps militaris was constructed by improved Agrobacterium tumefaciens-mediated transformation and screened for degradation features. Six mutants with altered characters in in vitro and in vivo fruiting body production, and cordycepin formation were found to contain a single copy T-DNA. T-DNA flanking sequences of these mutants were identified by thermal asymmetric interlaced-PCR approach. ATP-dependent helicase, cytochrome oxidase subunit I and ubiquitin-like activating enzyme were involved in in vitro fruiting body production, serine/threonine phosphatase involved in in vivo fruiting body production, while glucose-methanol-choline oxidoreductase and telomerase reverse transcriptase involved in cordycepin formation. These genes were analyzed by bioinformatics methods, and their molecular function and biology process were speculated by Gene Ontology (GO) analysis. The results provided useful information for the control of culture degeneration in commercial production of C. militaris.

Optimization of Conditions for In Vitro Germination of Seeds of Couroupita guianensis Aubl. - A Threatened Tree with Recalcitrant Seeds

  • M., Manokari;Shekhawat, Mahipal S.
    • Journal of Forest and Environmental Science
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    • v.34 no.5
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    • pp.388-394
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    • 2018
  • An efficient and reproducible protocol for in vitro germination of seeds of Couroupita guianensis has been developed in the present study. Couroupita guianensis is a medicinally important tree and categorized as threatened plant species by the IUCN. The seeds were surface sterilized using 0.1% $HgCl_2$ and inoculated on MS (Murashige and Skoog) and Woody Plants (Llyod and McCown) medium. The seeds with partially removed seed coat responded better (100%) on half strength MS medium as compared to the full (51%), one-fourth strength MS (79%) and the WP medium. Half strength MS medium conjunct with $2.0mg{\cdot}L^{-1}$ indole-3 butyric acid (IBA) was found best among the different types of auxins and cytokinins tested for seed germination, as all the seeds germinated on this medium combination. The seedlings were successfully hardened in the greenhouse and restored in the field with 83% survival rate. The protocol can be used as conservation measure for large scale multiplication of this threatened forest tree species.

USE OF COW FAECES AT DIFFERENT TIMES AFTER BEING VOIDED AS A SOURCE OF MICRO-ORGANISMS IN IN VITRO DIGESTIBILITY ASSAYS OF FORAGES

  • Akhter, S.;Owen, E.;Hossain, M.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.4
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    • pp.371-374
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    • 1996
  • Experiments were conducted to determine the effect of time intervals between collecting and use of cattle faeces as a source of micro-organisms in in vitro digestibility assays of forages. The results suggested that temperature conservation capacity by faeces depended on the size of the sample. There was no significant difference(p>0.05) between the first (T1 or 08:30 h) and second using time (T2 or 10:30 h). In vitro organic matter digestibility was significantly lower when faeces was used 5 h (T3 or 13:30 h) after collection. However, the organic matter digestibility determined at the second using time (T2) and third using time (T3) were highly correlated ($R^2=0.99$) with the first using time. It was concluded that faeces can be used as a source of microorganisms for in vitro digestibility assays of forages even 5 h after being voided.

Regeneration of Cryopreserved Pear Shoot Tips Grown in Vitro by Encapsulation-Dehydration

  • Yi, JungYoon;Lee, YoungYi;Lee, GiAn;Son, EunHo;Park, HongJae
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.612-617
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    • 2017
  • The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro-grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in "BaeYun No. 3" (55.7%) and "Whanggeum" (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro-grown plantlets of pear germplasm.

Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells

  • Feng, Wanyou;Chen, Shibei;Do, Dagiang;Liu, Qinyou;Deng, Yanfei;Lei, Xiaocan;Luo, Chan;Huang, Ben;Shi, Deshun
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1407-1415
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    • 2016
  • Isolation and culture of spermatogonial stem cells (SSCs) are attractive for production of genetic modified offspring. In the present study, buffalo spermatogonial stem-like cells were isolated, cultured and expression pattern of different germ cell marker genes were determined. To recover spermatogonia, testes from age 3 to 7 months of buffalo were decapsulated, and seminiferous tubules were enzymatically dissociated. Two types of cells, immature sertoli cell and type A spermatogonia were observed in buffalo testes in this stage. Germ cell marker genes, OCT3/4 (Pou5f1), THY-1, c-kit, PGP9.5 (UCHL-1) and Dolichos biflorus agglutinin, were determined to be expressed both in mRNA and protein level by reverse transcription polymerase chain reaction and immunostaining in buffalo testes and buffalo spermatogonial stem-like cells, respectively. In the following, when the isolated buffalo buffalo spermatogonial stem-like cells were cultured in the medium supplemented 2.5% fetal bovine serum and 40 ng/mL glial cell-derived neurotrophic factor medium, SSCs proliferation efficiency and colony number were significantly improved than those of other groups (p<0.05). These findings may help in isolation and establishing long term in vitro culture system for buffalo spermatogonial stem-like cells, and accelerating the generation of genetic modified buffaloes.

Induction of in vitro root tubers in Holostemma annulare (Roxb.) K. Schum. for the production of bioactive metabolites

  • Smitha Devi, Padmavathi Amma Somasekharan Nair;Hemanthakumar, Achuthan Sudarsanan;Preetha, Thankappan Suvarna
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.230-239
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    • 2022
  • Holostemma annulare (Family Asclepiadaceae) is an invaluable vulnerable medicinal plant; the root tubers are used in Ayurveda medicine and by folk healers to treat various ailments. In this study, Schenk and Hildebrandt medium fortified with the cytokinins 6-benzyl adenine, kinetin, and auxins, including indole 3-butyric acid, indole 3-acetic acid, α-naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic acid, were checked for their efficiency on root tuber induction from different explants. Adventitious root tubers were more successfully induced from in vitro leaf segments and shoots when cultured in Schenk and Hildebrandt medium supplemented with 0.5 mg/l of α-naphthaleneacetic acid. In addition, preliminary phytochemical analysis of in vitro root tubers and identification of different secondary metabolites were conducted. Thin layer chromatography and high performance thin layer chromatography analysis of the crude methanolic extracts of the in vitro root tuber identified the presence of lupeol, a bioactive triterpene. Adventitious root tuber induction offers a novel method for the in vitro production of bioactive metabolites that can be scaled up by bioreactors, thus ensuring the conservation and sustainable utilization of H. annulare. The study warrants further scale-up production and pharmacological investigation that can be extended for pharmaceutical needs.

Screening of aldose reductase inhibitory activities of Korean folk plants in Chungcheong Province

  • Lee, Dong Gu;Lee, Ki Ho;Choi, Kyung;Ku, Jajung;Park, Kwang-Woo;Cho, Eun Ju;Lee, Sanghyun
    • Korean Journal of Agricultural Science
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    • v.40 no.1
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    • pp.47-51
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    • 2013
  • To search for the aldose reductase (AR) inhibitors from Korean folk plants, the inhibition of rat lens AR in vitro using the methanol (MeOH) extracts from Korean folk plants in Chungcheong Province was investigated. Among Korean folk plants tested, the MeOH extract of Persicaria longiseta showed highest inhibition of AR ($IC_{50}$ value, $5.14{\mu}g/ml$). Consequently, P. longiseta has a possibility of new natural resources for the development of AR inhibitor for the prevention of diabetic complications.

Micropropagation of Echinosophora koreensis Nakai, a Korean Endemic Species in Danger, Using Axillary Buds

  • Hyunseok Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.60-60
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    • 2020
  • To establish in vitro axillary bud culture conditions of Echinosophora koreensis Nakai, one of Korean endemic endangered species famous for beautiful flowers, we tested the influence of plant growth regulators (PGRs) in shooting and rooting stage from in vitro plants. In shoot multiplication, addition of 6-benzylaminopurine (BA) to the media induced 2.5 to 3 shoots per bud during 4 weeks of culture. And media including 0.5 mg L-1 thidiazuron (TDZ) produced 3 to 4 shoots per bud. However, zeatin and isopentenyl adenine (2-ip) were not successful to increase shoot number, and the combination treatments of BA with other PGRs were also not effective. Shoots were smaller than 2 cm in length, in most of the treatments. In rooting, naphthalene acetic acid (NAA) treatments in the range of 0.5 to 4.0 mg L-1 appeared to increase rooting rate by 10% to 60% approximately when compared with the control but roots developed with callus clusters. Indole butyric acid (IBA) addition had little effect on rooting (below 10%), while some roots were longer than in NAA treatments and some shoots were longer on high IBA concentrations (4.0 to 8.0 mg L-1). It is suggested that micropropagation is a highly applicable and promising to multiplication and conservation of rare and endangered endemic species.

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Inhibition of Aldose Reductase from Rat Lenses by Methanol Extracts from Korean Folk Plants

  • Kim, Hye-Min;Mok, So-Youn;Lee, Jeong-Min;Cho, Eun-Ju;Choi, Kyung;Ku, Ja-Jung;Park, Kwang-Woo;Lee, Sang-Hyun
    • Natural Product Sciences
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    • v.16 no.4
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    • pp.285-290
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    • 2010
  • The inhibition of rat lens aldose reductase (AR) in vitro using methanol extracts from Korean folk plants was investigated. Among them, the extracts of Saussurea grandifolia and Rumex crispus showed highest inhibition of AR. $IC_{50}$ values of the extracts from S. grandifolia and R. crispus were demonstrated 0.07 and 0.05 mg/ml, respectively. Korean folk plants such as S. grandifolia and R. crispus has a possibility of new natural resources for the inhibition of AR.