WO2007069643A1 - 粉体を用いて形質転換効率を向上させる方法 - Google Patents
粉体を用いて形質転換効率を向上させる方法 Download PDFInfo
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- WO2007069643A1 WO2007069643A1 PCT/JP2006/324839 JP2006324839W WO2007069643A1 WO 2007069643 A1 WO2007069643 A1 WO 2007069643A1 JP 2006324839 W JP2006324839 W JP 2006324839W WO 2007069643 A1 WO2007069643 A1 WO 2007069643A1
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- WO
- WIPO (PCT)
- Prior art keywords
- powder
- plant
- plant material
- agrobacterium
- gene
- Prior art date
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
- C12N15/8205—Agrobacterium mediated transformation
Definitions
- the present invention relates to a method for efficiently introducing a gene into a plant material via a bacterium belonging to the genus Agrobacterium.
- the gene transfer method using bacteria belonging to the genus Agrobacterium is universally used as a method for transforming dicotyledonous plants.
- the host of agrobacterium was limited to dicotyledonous plants and not to monocotyledonous plants (De Cleene, M. and De Ley, J., (1 976) Bot. Rev.
- Non-Patent Document 24 Hoekema, A., et al., (1983) Nature, 303: 179-180
- the presence of powder means a state in which powder is present when inoculating plant material with Agrobataterium bacteria. Therefore, after the Agrobataterium bacterial suspension is premixed with the powder, the mixture may be inoculated into the plant material; after the plant material is premixed with the powder, the mixture Terium bacteria may be inoculated; or plant material may be inoculated with agrobacterium at the same time by mixing agar butterium suspension, powder, and plant material at the same time May be. In the present invention, it is sufficient to mix so as to be moderately homogeneous, and it is not necessary to stir strongly. For example, when mixing using a device that stirs with a relatively strong force such as a vortex mixer, the mixing is performed for a short mixing time, for example, 1 minute or less, preferably 45 seconds or less, more preferably 30 seconds or less.
- a desired gene to be introduced into a plant can be incorporated into a restriction enzyme site in the T DNA region of the plasmid by a conventional method, and PPT (phosphinoslysin) incorporated simultaneously or separately into the plasmid.
- PPT phosphinoslysin
- selection can be made based on an appropriate selection marker such as a gene having resistance to drugs such as hygromycin, kanamycin, and paromomycin. Larger ones with many restriction sites may not always be easy to introduce the desired DNA into the T-DNA region using conventional subcloning techniques.
- the target DNA can be introduced by utilizing the homologous recombination in the cells of a bacterium belonging to the genus Agrobacterium by the three-line hybridization method.
- the size of the introduced gene is preferably about lOObp to 200 kbp.
- step (c) infecting the plant material prepared in step (a) with the agrobacterium prepared in (b) in the presence of powder;
- the present invention provides a method for gene transfer and transformation of a plant by the agrobacterium method with high gene transfer efficiency and transformation efficiency.
- phosphate buffer pH 6.8 containing 0.1% Triton X-100 and allowed to stand at 37 ° C. for 1 hour. After removal of agrobatterium with phosphate buffer, add phosphate buffer containing 1. OmM 5 bromo 4 black mouth 3-indolinole- ⁇ -D glucuronic acid (X-Glue) and 20% methanol. Added. After treatment at 37 ° C for 24 hours, the structure showing blue color was observed under a microscope.
- the immature embryos after co-cultivation were cultured in a medium containing PPT, and the obtained callus was placed on a re-seed medium containing PPT and cultured.
- GUS analysis was performed on PPT-resistant plants that had been subdivided. GUS-positive plants were obtained from both immature embryos, but in both inoculated and inoculated seeds, the immature embryos inoculated with powder added to the inoculation source were compared to the immature embryos with no powder added compared to the control immature embryos to which no powder was added. It was clear that the transformation efficiency was improved by adding powder (Table 1).
- HA hydroxyapatite
- S G silica gel
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Physical Or Chemical Processes And Apparatus (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Inorganic Compounds Of Heavy Metals (AREA)
Abstract
Description
Claims
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DK06834594.1T DK1964919T3 (da) | 2005-12-13 | 2006-12-13 | Fremgangsmåde til forbedring af transformationseffektivitet ved anvendelse af pulver |
CN2006800469361A CN101331227B (zh) | 2005-12-13 | 2006-12-13 | 使用粉体来提高转化效率的方法 |
EP06834594A EP1964919B1 (en) | 2005-12-13 | 2006-12-13 | Method for improving transformation efficiency using powder |
JP2007550198A JP5260963B2 (ja) | 2005-12-13 | 2006-12-13 | 粉体を用いて形質転換効率を向上させる方法 |
AU2006324560A AU2006324560B2 (en) | 2005-12-13 | 2006-12-13 | Method for Improving Transformation Efficiency Using Powder |
AT06834594T ATE545700T1 (de) | 2005-12-13 | 2006-12-13 | Verfahren zur verbesserung der transformationseffizienz unter verwendung eines pulvers |
US12/086,426 US8324456B2 (en) | 2005-12-13 | 2006-12-13 | Method for improving transformation efficiency using powder |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JPPCT/JP2005/022863 | 2005-12-13 | ||
PCT/JP2005/022863 WO2007069301A1 (ja) | 2005-12-13 | 2005-12-13 | 粉体を用いて形質転換効率を向上させる方法 |
Publications (1)
Publication Number | Publication Date |
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WO2007069643A1 true WO2007069643A1 (ja) | 2007-06-21 |
Family
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Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2005/022863 WO2007069301A1 (ja) | 2005-12-13 | 2005-12-13 | 粉体を用いて形質転換効率を向上させる方法 |
PCT/JP2006/324839 WO2007069643A1 (ja) | 2005-12-13 | 2006-12-13 | 粉体を用いて形質転換効率を向上させる方法 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/JP2005/022863 WO2007069301A1 (ja) | 2005-12-13 | 2005-12-13 | 粉体を用いて形質転換効率を向上させる方法 |
Country Status (7)
Country | Link |
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US (1) | US8324456B2 (ja) |
EP (1) | EP1964919B1 (ja) |
CN (1) | CN101331227B (ja) |
AT (1) | ATE545700T1 (ja) |
AU (1) | AU2006324560B2 (ja) |
DK (1) | DK1964919T3 (ja) |
WO (2) | WO2007069301A1 (ja) |
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WO2009122962A1 (ja) | 2008-03-31 | 2009-10-08 | 日本たばこ産業株式会社 | アグロバクテリウム菌による形質転換植物の作成方法 |
WO2011013764A1 (ja) | 2009-07-29 | 2011-02-03 | 日本たばこ産業株式会社 | アグロバクテリウム菌を用いた、コムギ属の植物へ遺伝子導入を行う方法、コムギ属の植物の形質転換植物の作成方法 |
WO2012015039A1 (ja) | 2010-07-29 | 2012-02-02 | 日本たばこ産業株式会社 | アグロバクテリウム菌を用いた、オオムギ属植物へ遺伝子導入を行う方法およびオオムギ属植物の形質転換植物の作成方法 |
WO2015085990A1 (de) | 2013-12-15 | 2015-06-18 | Kws Saat Ag | Selektionsmarker-freies rhizobiaceae-vermitteltes verfahren zur herstellung einer transgenen pflanze der gattung triticum |
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US10568328B2 (en) | 2013-03-15 | 2020-02-25 | Monsanto Technology Llc | Methods and compositions for weed control |
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- 2006-12-13 AT AT06834594T patent/ATE545700T1/de active
- 2006-12-13 AU AU2006324560A patent/AU2006324560B2/en active Active
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- 2006-12-13 EP EP06834594A patent/EP1964919B1/en active Active
- 2006-12-13 CN CN2006800469361A patent/CN101331227B/zh active Active
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- 2006-12-13 DK DK06834594.1T patent/DK1964919T3/da active
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Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009122962A1 (ja) | 2008-03-31 | 2009-10-08 | 日本たばこ産業株式会社 | アグロバクテリウム菌による形質転換植物の作成方法 |
US8357836B2 (en) | 2008-03-31 | 2013-01-22 | Japan Tobacco Inc. | Agrobacterium-mediated method for producing transformed maize or rice |
AU2009232967B2 (en) * | 2008-03-31 | 2015-04-02 | Kaneka Corporation | Agrobacterium-mediated method for producing transformed plant |
WO2011013764A1 (ja) | 2009-07-29 | 2011-02-03 | 日本たばこ産業株式会社 | アグロバクテリウム菌を用いた、コムギ属の植物へ遺伝子導入を行う方法、コムギ属の植物の形質転換植物の作成方法 |
US8884101B2 (en) | 2009-07-29 | 2014-11-11 | Japan Tobacco Inc. | Method of gene introduction into Triticum plant using Agrobacterium, and a method of producing transformed Triticum plant |
JP5766605B2 (ja) * | 2009-07-29 | 2015-08-19 | 日本たばこ産業株式会社 | アグロバクテリウム菌を用いた、コムギ属の植物へ遺伝子導入を行う方法、コムギ属の植物の形質転換植物の作成方法 |
WO2012015039A1 (ja) | 2010-07-29 | 2012-02-02 | 日本たばこ産業株式会社 | アグロバクテリウム菌を用いた、オオムギ属植物へ遺伝子導入を行う方法およびオオムギ属植物の形質転換植物の作成方法 |
WO2015085990A1 (de) | 2013-12-15 | 2015-06-18 | Kws Saat Ag | Selektionsmarker-freies rhizobiaceae-vermitteltes verfahren zur herstellung einer transgenen pflanze der gattung triticum |
DE102013020605A1 (de) | 2013-12-15 | 2015-06-18 | Kws Saat Ag | Selektionsmarker-freies rhizobiaceae-vermitteltes verfahren zur herstellung einer transgenen pflanze der gattung triticum |
Also Published As
Publication number | Publication date |
---|---|
EP1964919A4 (en) | 2009-05-27 |
CN101331227A (zh) | 2008-12-24 |
AU2006324560A1 (en) | 2007-06-21 |
EP1964919B1 (en) | 2012-02-15 |
AU2006324560B2 (en) | 2013-08-29 |
DK1964919T3 (da) | 2012-05-14 |
US8324456B2 (en) | 2012-12-04 |
WO2007069301A1 (ja) | 2007-06-21 |
CN101331227B (zh) | 2012-03-28 |
US20110131685A1 (en) | 2011-06-02 |
ATE545700T1 (de) | 2012-03-15 |
EP1964919A1 (en) | 2008-09-03 |
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