• Title/Summary/Keyword: leaf explants

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Development of a Method for High throughput Screening of Antagonistic Substances against Rice Pathogens using Rice Leaf Explants (벼의 생엽절편을 이용한 병원균 억제물질의 대량 스크리닝 방법 개발)

  • Park, Sait-Byul;Lee, Choong-Hwan;Kim, Tae-Jong;Kang, Lin-Woo;Lee, Byoung-Moo;Kim, Jeong-Gu
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.39-42
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    • 2012
  • A new method for the high throughput screening of antagonistic substances against rice pathogens using rice leaf explants was developed. This method can be used to confirm the activities of any compound or mixture suppressing rice bacterial blight (BB) before field tests. Xanthomonas oryzae pv. oryzae (Xoo) culture medium was distributed in 96 well plates with equally sized explants and the active compounds were added to the wells. The strength suppressing BB was converted into an area percent of the lesion on the rice explants. The explants under BB suppressing activity remained uninfected maintaining their actual green color, while infected explants exhibited pale yellow-colored lesions. Based on the results, this method seems to be faster and easier, dose-dependent, and can be performed all-at-once with a small amount of unspecified compounds. This method also has the potential to be applied to inspection activities for the suppression of other waterborne crop diseases.

Plant Regeneration from Floral Stem Cultures of Nymphoides indica (L.) O. Kuntze. via Somatic Embryogenesis (어리연꽃 (Nymphoides indica (L.) O. Kuntze) 화경 배양으로부터 체세포배발생을 통한 식물체 재생)

  • Oh, Myung-Jin;Min, Sung-Ran;Liu, Jang-Ryol;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.7-10
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    • 2007
  • Plant regeneration system from floral stem of Mymphoides indica via somatic embryogenesis was established. After four weeks of culture onto 1/2MS medium containing 2,4-D, pale-yellow globular structures and calluses were formed on the cut surface of floral stem explants. Upon transfer to 1/2MS basal medium, pale-yellow globular structures were developed into somatic embryos and normal plantlets. These results indicated that pale-yellow globular structures and calluses from floral stem were globular embryos and embryogenic calluses, respectively. The frequency of embryogenic callus formation from floral stem was reached to nearly 100% when floral stem was cultured onto 1/2Ms medium supplemented with low concentration of 2,4-D (0.1 to 0.3 mg/L). However, the higher concentration of 2,4-D resulted in decrease of the frequency of embryogenic callus formation. In this study, low concentration of 2,4-D had a stimulative role in embryogenic callus formation, whereas BA showed inhibitory role in callus formation. In comparison to floral stem, leaf explants showed low frequency of embryogenic callus formation. The highest frequency of embryogenic callus formation from leaf explants was 9.5% when leaf explants were cultured onto 1/2MS medium supplemented with 0.3 mg/L of 2,4-D. The plant regeneration system of Nymphoides indica established in this study, might be applied to mass proliferation, conservation of genetic resources and genetic transformation for molecular breeding.

Plant regeneration from suspension-cultured cell clusters of Arabidopsis thaliana (애기장대(Arabidopsis thaliana)의 현탁배양세포괴로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.195-200
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    • 1998
  • Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

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Comparative Effect of Plant Growth Regulators on Callus Induction and Plant Regeneration in Glehnia littoralis Schmidt et Miquiel (갯방풍(Glehnia littoralis Schmidt et Miquiel)에서 캘러스 유도 및 식물체 재분화에 미치는 생장조절제의 효과)

  • 이병국;한미숙;정양균;나의식;윤성중;유남희
    • Korean Journal of Plant Resources
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    • v.17 no.2
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    • pp.153-160
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    • 2004
  • Glehnia littoralis is known as an edible and medicinal plant using green loaves and mature roots of plant. In the present paper, the influence of plant growth regulators on callus induction and plant regeneration was investigated. Callus induction and regeneration occurred from leaf and petiole explants in Glehnia littoralis. Optimal condition of plant growth regulators for callus induction from leaf and petiole explants was MS basal medium supplemented with 2mg/L 2,4-D and 2mg/L BA. The frequency of callus induction was higher in petiole explant than leaf. When the callus was cultured on MS basal medium supplemented with 0∼1 mg/L IAA, 0∼1mg/L NAA and 0∼2mg/L BA for about 65 days, the most effective plant growth regulators on plant regeneration from callus were 1mg/L NAA and 2mg/L BA. The plantlets acclimatized successfully and grown in vermiculite matrix.

Somatic Embryogenesis from Various Parts of Muscari comosum var. plumosum

  • Xudong He;Ko Jeong-Ae;Choi Jeong-Ran;Kim Hyung-Moo;Kim Myung-Jun;Choi So-Ra;Kim Young-Gon;Kim Dong-Hee;Kim Hyun-Soon
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.427-431
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    • 2006
  • In vitro high-frequency plant regeneration of Muscari comosum var. plumosum through somatic embryogenesis was obtained via two developmental pathways: direct embryos and multiple shoots regenerated from embryogenic callus. Flower bud with pedicel, receptacle, petal and ovary wall, floral stalk and leaf as explants were cultured in MS medium supplemented with various plant growth regulators. Embryos formed directly from pedicel, receptacle and floral stalk. Depending on explant sources, the optimal medium was MS medium supplemented with 0.2 mg/L IBA and 0.3 mg/L BA, 3.0 mg/L IBA and 3.0 mg/L BA, and MS-free medium for pedicel, receptacle, and floral stalk, respectively. Multiple shoots regenerated from embryogenic cal]i which was initiated from petal, ovary and leaf were observed in MS medium with different concentrations and combinations of hormone. The most suitable medium for each type of explant was 3.0 mg/L IBA and 3.0 mg/L BA(petal and ovary) and 5.0 mg/L IBA and 5.0 mg/L BA (leaf) Furthermore, the combination of 0.1 mg/L 2,4-D and 1.0 mg/L BA was also good for all sources of explants not only for direct embryo formation, but also, for embryogenic callus induction.

Efficient transformation of Actinidia arguta by reducing the strength of basal salts in the medium to alleviate callus browning

  • Han, Meili;Gleave, Andrew P.;Wang, Tianchi
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.129-138
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    • 2010
  • An efficient transformation system for high-throughput functional genomic studies of kiwifruit has been developed to overcome the problem of necrosis in Actinidia arguta explants. The system uses Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pART27-10 to inoculate leaf strips. The vector contains neomycin phosphotransferase (nptII) and ${\beta}$-glucuronidase (GUS) (uidA) genes. A range of light intensities and different strengths of Murashige and Skoog (MS) basal salt media was used to overcome the problem of browning and/or necrosis of explants and calli. Callus browning was significantly reduced, resulting in regenerated adventitious shoots when the MS basal salt concentration in the culture medium was reduced to half-strength at low light intensity ($3.4\;{\mu}mol\;m^{-2}\;s^{-1}$) conditions. Inoculated leaf strips produced putative transformed shoots of Actinidia arguta on half-MS basal salt medium supplemented with 3.0 $mg\;l^{-1}$ zeatin, 0.5 $mg\;l^{-1}$ 6-benzyladenine, 0.05 $mg\;l^{-1}$ naphthalene acetic acid, 150 $mg\;l^{-1}$ kanamycin and 300 $mg\;l^{-1}$ $Timentin^{(R)}$. All regenerated plantlets were deemed putativ transgenic by histochemical GUS assay and polymerase chain-reaction analysis.

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.

Effect of Morphological Factors, Antibiotics and Agrobacterium Co-cultivation in the Efficiency of Somatic Embryogenesis of Eggplant (Solanum melongena L.)

  • Claudia Magioli;Erika Tarre;Krul, William-Robert;Elisabeth Mansur
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.19-25
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    • 2001
  • Induction of somatic embryogenesis from Brazilian eggplant variety F-100 was studied in response to four auxin types. NAA, at the optimal concentration of 54 $\mu\textrm{m}$, was the only one that resulted in the induction of somatic embryos in either leaf and cotyledon explant and, at murk lower intensity and frequency, in hypocotyl and epicotyl explants. The optimal temperatures for embryo induction were 28 and 35$^{\circ}C$ for cotyledon and leaf explants. Incubation at 22$^{\circ}C$ caused a significant reduction both in the frequency and intensity of induction. This system was used to study the effects of position and orientation of the tissue on the culture medium as well as of antibiotics and explant co-cultivation with Agrobacterium on the efficiency of somatic embryo induction. The intensity of embryo induction was greater in the midsections of cotyledons relative to apical and basal regions, when the abaxial surface was in contact with the culture medium. The presence of antibiotics resulted in approximately 40-60% reduction of embryo induction relative to control explants, which originated 335$\pm$26.6 embryos. Co-cultivation with Agrobacterium before treatment with antibiotics caused a more drastic reduction (80-99%). Ampicilin treatment after cocultivalion with Agrobacterium caused the least inhibitory effect, allowing the production of 60 embryos/explant.

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Effect of plant growth regulators on plant regeneration from the Sedum rotundifolium D. Lee (둥근잎꿩의비름(Sedum rotundifolium D. Lee)의 식물체 재분화에 미치는 식물생장조절제의 영향)

  • Kwon, Hye-Kyoung;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.84-88
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    • 2010
  • To establish the system of In vitro plant regeneration, the floral bud and leaf explants of Sedum rotundifolium were cultured on the MS media supplemented with different concentration of 2,4-D, NAA, and BA. The callus induction was more effective in the floral explants than the leaf explants, and was the best on MS medium containing 1.0 or 2.0 mg/L 2,4-D and 1.0 mg/L BA. The highest numbers of shoots were regenerated when callus were cultured on MS medium containing 2.0 mg/L 2,4-D and 1.0 mg/L BA for 8 weeks. The normal root formation from shoot was effective on the MS medium containing IAA alone. The regenerated plantlets were transferred to the pot and acclimatized successfully.

Plant regeneration from callus of Iris odaesanensis Y. N. Lee native to Korea via organogenesis

  • Bae, Kee-Hwa;Yoo, Kyoung-Hwa;Lee, Mi-Hyun;Jeong, Jae-Hun;Choi, Yong-Eui;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.40 no.3
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    • pp.163-168
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    • 2013
  • Iris odaesanensis Y. N. Lee. is an important endangered and native plant belonging to the family Iridaceae in Korea. This study describes a method for rapid micropropagation of this species via from leaf, rhizome and root explants derived calli. Leaf, rhizome and root explants were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) for callus induction. Rhizome explants yielded calli at a frequency of 72% when cultured at 1.0 mg/l 2,4-D. Calli were maintained at 1.0 mg/l 2,4-D. These calli were transferred to MS medium supplemented with 0, 0.5, 1.0, and 2.0 mg/l 2,4-D in combination with 0, 0.5, 1.0, and 3.0 mg/l BA for adventitious shoot induction. The highest number of adventitious shoot (228.9 per petri-dish) were formed at 1.0 mg/l 2,4-D and 1.0 mg/l BA. WPM medium was the best to convert calli into plantlets, where up to 98.2% of calli were regenerated into plantlets. This in vitro propagation protocol should be useful for conservation of this endangered plant.