• Title/Summary/Keyword: rice callus

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Transformation of Rice Embryogenic Cells by Electroporation Mediated Plasmid Uptake into Protoplasts 1. Plant Regeneration from Electroporated Protoplasts of Rice (원형질체 내 Plasmid Electroporation에 의한 벼 배발생세포의 형질전환 1. 벼의 Electroporation 원형질체로부터 식물체 재분화)

  • 김명덕;최성진김준철
    • KSBB Journal
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    • v.10 no.1
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    • pp.23-29
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    • 1995
  • Calli were induced from leaf base region of germinated rice(Oryza sativa L. cv. Nakdong) with high frequency of up to 65% on LS medium supplemented with $2.5mg/{\ell}2$, 4-D in the dark at $27^{\circ}C$. Embryogenic calli of pale yellow, globular type were selected and used for the initiation of cell suspension cultures in AA2 liquid medium with $2mg/\ell$ 2,4-D, 0.2mg/$\ell$ kinetin arid $0.1mg/\ell$ GA3. Protoplasts were isolated from the embryogenic cell suspensions after 4 months of culture and then were electroporated with 400V/cm for 1 msec. Electroporated protoplasts divided with plating efficiency of 1.1% on PCM liquid medium supplemented with $2.5mg/\ell$ 2, 4-D, $0.1mg/\ell$ kinetin and 10mM proline. The protoplasts-derived microcalli were cultured on $0.2{\mu}m$ membrane fitter placed onto LS2.5 solid medium containing fine suspension cells as a feeder cells, for 2 weeks in the dark at $27^{\circ}C$. After an additional 2 weeks of culture under fluorescent light of $30{\pm}/3{\mu}E$.m^{-2}S^{-1}, yellow calli of 2mm diameter were transferred to regeneration medium. Shoots were produced from the green spot of protoplasts-derived calli and plants were regenerated form protoplast-derived green calli with frequencies of 11∼33%.

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Increment of fructan biosynthesis in rice by transformation of 1-sst and 1-fft genes isolated from jerusalem artichoke (Helianthus tuberosus L.) (돼지감자 유래 1-sst와 1-fft 유전자의 형질전환 발현에 의한 벼의 fructan 생합성 증진)

  • Kang, Kwon-Kyoo;Song, Beom-Heon;Lee, Gyong-A;Lee, Hye-Jung;Park, Jin-Ha;Jung, Yu-Jin;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.102-109
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    • 2010
  • Fructan has been found to accumulate in various tissues during periods when light levels increased carbon fixation where low temperatures reduced growth rates while photosynthesis continued. In this study, we have cloned 1-sucrose:sucrose fructosyl transferase(1-sst) and 1-fructan: fructan fructosyl transferase (1-fft, a key enzyme for the synthesis of fuctan) from Jerusalem Artichoke (Helianthus tuberosus L.). The recombinant vector with 1-sst and 1-fft has been constructed under the control of 35S promoter of KJGV-B2 vector and transgenic plants obtained by Agrobacterium tumefaciens LBA4404. PCR analysis carried out on the putative transgenic plants for amplification of the coding region of specific gene (1-sst, 1-fft), and HPT genes. Transgenic lines carrying of 1-sst and 1-fft were confirmed for integration into the rice genome using Southern blot hybridization and RT-PCR. The transgenic plants in $T_2$ generation were selected and expression pattern analysis revealed that 1-sst and 1-fft were stable. This analysis confirmed the presence of low-molecular-weight fructan in the seedling of the transgenic rices. Therefore, cold tolerance and carbohydrate metabolism will be possible to develop resistant plants using the transgenic rice.

Proteome Profiling Unfurl Differential Expressed Proteins from Various Explants in Platycodon Grandiflorum

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Cho, Kab-Yeon;Boo, Hee-Ock;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.97-106
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    • 2015
  • Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}$ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

Herbicidal and Insecticidal Potentials of 5-Aminolevulinic acid, a Biodegradable Substance (생분해성 생리활성물질 5-aminolevulinic acid의 제초 및 살충활성)

  • Chon, Sang-Uk
    • The Korean Journal of Pesticide Science
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    • v.11 no.1
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    • pp.52-58
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    • 2007
  • ALA (5-aminolevulinic acid) has been proposed as a tetrapyrrole-dependent photodynamic herbicide and insecticide by the action of the protoporphyrinogen IX oxidase (Protox IX). The present study was conducted to determine growth responses of plant and insects to ALA, biodegradable biopesticidal substance. In the paddy condition experiment, plant height and shoot fresh weight of barnyardgrass (Echinochloa crus-galli) was more reduced by ALA than rice plants, even though both plant species show great phytotoxicity. Hairy crabgrass (Digitaria sanguinalis), a monocot weed, was more sensitive to ALA at 5mM under upland condition when ALA applied on the foliage, compared with soybean (Glycine max) as a dicot crop. ALA solutions were tested for their insecticidal and larvicidal activities against Spodaptera exigua (Hubner) and Tetranychus urticae Koch. by foliar application and leaf-dipping method. The result showed higher insecticidal activity of ALA at 10mM and its mixture with insecticide luferon against S. exigua. Strongest insecticidal activity against T. urticae was observed from the ALA solution at 10mM 72 days after application. This results show that ALA solution had potent herbicidal and insecticidal activities against agricultural pests even though their activities were lower than those of synthetic pesticides.

Secondary Products in Cell Suspension Culture of Salix koreensis (버드나무(Salix koreensis) 현탁배양(懸濁培養) 세포(細胞)의 대사산물(代謝産物))

  • Park, Young Goo;Shin, Dong Ill;Lee, Sang Goo
    • Journal of Korean Society of Forest Science
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    • v.78 no.2
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    • pp.198-208
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    • 1989
  • Cell suspension cultures for Salix koreensis was well established at the supplements of 2, 4-D with cytokinin particulary the combination of 1.0 mg/l 2, 4-D with 0.1 mg/l of zeatin. These combined rates of phytohormones are also effective to callus induction from S, koreensis leaf and its multiplication. Cultured media exhibited the great inhibitory effect on the germination of rice, barnyard grass and lettuce seeds, indicating the presence of biologically active substances in media. Several phenolic compounds such as pyrogallol, sinapic acid, cinnamic acid, tannic + gallic and p-chlorobenzoic acid were detected in the cell suspension culture. The inhibitory effect exhibited by cultured media may be partly attributed to these phenalic compounds.

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Isolation and Characterization of Cyclophilin 1 (ClCyP1) Gene from Codonopsis lanceolata (더덕의 주근에서 유래한 Cyclophilin 1 (ClCyP1) 유전자의 분리 및 분석)

  • 양덕춘;이강;인준교;이범수;김종학
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.239-247
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    • 2004
  • A cyclophilin 1 cDNA clone(GenBank accession no.CF924191) was isolated from the taproot of C. lanceolata and designed as C1CyP1. Determination of the nucleotide sequence of C1CyPl identified an open reading frame of 525bp, which shared high homologies with cyclophilins that were previously reported in other organisms. The C1CyP1 amino acid sequence possesses 7 amino acid residue stretch(KSGKPLH) that is characteristic of plant cytosolic dehydrins. Currently available amino acid residues of plant cyclophilins were compared to examine their phylogenetic relationship to C1CyP1. In the phylogenetic analysis, based on the aligned sequences, C1CyP1 showed high homology with arabidopsis ROC2 and rice CyP1. The transcript that corresponded to C1CyP1 was abundant in callus, but only basal level of transcript was detected in stem, leaf and root. For the study in the defense mechanism against various stresses, we report expression patterns of this gene by quantative RT-PCR.

The Filter Membrane Culture Procedure with Feeder Cells in Rice Protoplast Culture (Filter membrane과 feeder세포를 이용한 벼의 원형질체 배양)

  • LEE, Sung-Ho;SHON, Young Geol;Lee, Soo In;DAVEY Micheal R.;COCKING Edward C.;CHO, Moo Je
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.295-303
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    • 1997
  • To investigate the response on feeder cell cultures, protoplasts isolated from cell suspensions initiated from mature seed scutellum-derived callus of the Japonica rice variety Taipei 309, were cultured on filter membranes under various conditions. The effects of various factors, such as gelling agents, feeder cell and protoplast densities, species of feeder cells and heat shock treatment, have been investigated to improve protoplast plating efficiencies on filter membranes. Maximum protoplast plating efficiencies were obtained when protoplasts were cultured on KPR medium semi-solidified with Sea Plaque agarose at a density of $5\;\times\;10^{5}\;ml^{-1}$ protoplasts in the presence of Lolium multiflorum as feeder cells (0.5 ml pcv per 10 ml of protoplast culture medium). Pre-culture heat shock treatments for 1 min. and 5 min. to the protoplasts did not give any appreciable increase on the plating efficiency of protoplasts in the presence of feeder cells. Maltose-supplemented medium was superior for plant regeneration from protoplast-derived colonies compared with medium containing only sucrose. The plants were transferred to the glasshouse, flowered and were fertile.

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The Studies of Activity of Retrotransposon(Tos17) according to Tissue Culture Periods in Rice(Oryza sativa L.) (벼 조직배양 기간에 따른 retrotransposon(Tos17)의 활성에 관한 연구)

  • Yang, Hee-Eun;Fang, Yilan;Shin, Young-Boum;Lee, Boung-Jin;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.20 no.5
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    • pp.389-397
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    • 2007
  • Using the active-increment of Tos17 copies in the genome of Oryza sativa L., there were many studies about induction and selection of new mutants. This study mainly focuses on the induction for retrotransposon(Tos17) activity in the callus of Ilpumbyeo(Oryza sativa L.) according to varied culture period and condition. The objectives of this study are obtaining various mutants($M_1$) through plant regeneration, identification of the mutation relation with Tos17, and subsequent phenotyping of the mutants($M_2,\;M_3$). A total of 371 $M_1$ mutants was obtained. The degree of Tos17 activity obtained regeneration plants with each different culture period was evaluated by Southern blot analysis. The result showed that control Ilpumbyeo rice has 5 numbers of copies and the band numbers obtained 7, 8, 9.5, 12, 6, 13.5, 17.5 from culture period of 1, 2, 3, 5, 6, 7, 8 month, respectively. In this study, the result showed that most effectual culture period for activity of Tos17 in Ilpumbyeo rice is 5 month. Hereafter, collections and analysis of various recombination plants will act on an important factor in multiplication and preservation of $M_2$ and $M_3$ generation. And an urgent and important subject is a development of screening method for selection of diverse mutants.