公开(公告)号：US07939328B1

公开(公告)日：2011-05-10

申请号：US08428238

申请日：1994-09-01

申请人： Hideaki Saito ,  Yuji Ishida ,  Yukoh Hiei ,  Toshihiko Komari

发明人： Hideaki Saito ,  Yuji Ishida ,  Yukoh Hiei ,  Toshihiko Komari

IPC分类号： C12N15/82

CPC分类号： C12N15/8205

摘要： Disclosed is a method of transforming monocotyledons which necessitates only a short period from the transformation to the regeneration of a whole plant as compared with the conventional methods, thus reducing the frequency of occurrence of mutants, and can be generally applied to the plants for which any system of regenerating the whole plants from protoplasts has not been established, and in which the material to be used can be readily prepared without any particular apparatuses. The present invention provides a method for transforming monocotyledons comprising transforming scutellum of an immature embryo of a monocotyledon with a bacterium belonging to genus Agrobacterium containing a desired gene, which immature embryo has not been subjected to a dedifferentiation treatment, to obtain a transformant.

摘要翻译： 本发明公开了一种转化单子叶植物的方法，其与常规方法相比仅需要从整个植物的转化到再生的短的时间，从而降低突变体的发生频率，并且通常可以应用于任何 尚未建立从原生质体再生整株植物的系统，其中待使用的材料可以在没有任何特定装置的情况下容易地制备。 本发明提供一种用于将单子叶植物的未成熟胚的鳞片状细菌与含有未成熟胚的未分化处理的所需基因的农杆菌属的农杆菌属转化成单转基因的方法，得到转化体。

公开(公告)号：US20100132068A1

公开(公告)日：2010-05-27

申请号：US12306163

申请日：2007-06-25

申请人： Yoshimitsu Takakura ,  Toshihiko Komari ,  Yuji Ishida ,  Toshiyuki Komori ,  Yukoh Hiei ,  Toshiki Mine ,  Teruyuki Imayama

发明人： Yoshimitsu Takakura ,  Toshihiko Komari ,  Yuji Ishida ,  Toshiyuki Komori ,  Yukoh Hiei ,  Toshiki Mine ,  Teruyuki Imayama

IPC分类号： A01H1/00 ,  C12N15/63

CPC分类号： C12N15/82 ,  C12N15/8205

摘要： The present invention aims to provide novel vectors for plant transformation.The vectors of the present invention are cosmid vectors having a full length of 15 kb or less characterized in that: 1) they contain an origin of replication of an IncP plasmid, but do not contain any origin of replication of other plasmid groups; 2) they contain the trfA1 gene of an IncP plasmid; 3) they contain an oriT of an IncP plasmid; 4) they contain the incC1 gene of an IncP plasmid; 5) they contain a cos site of lambda phage and the cos site is located outside the T-DNA; 6) they contain a drug resistance gene expressed in E. coli and a bacterium of the genus Agrobacterium; 7) they contain a T-DNA right border sequence of a bacterium of the genus Agrobacterium; 8) they contain a T-DNA left border sequence of a bacterium of the genus Agrobacterium; 9) they contain a selectable marker gene for plant transformation located between 7) and 8) and expressed in a plant; and 10) they contain restriction endonuclease recognition site(s) located between 7) and 8) for cloning a foreign gene.

摘要翻译： 本发明旨在提供用于植物转化的新型载体。 本发明的载体是全长为15kb或更小的粘粒载体，其特征在于：1）它们含有IncP质粒的复制起点，但不含有其他质粒组的任何复制起点; 2）它们含有IncP质粒的trfA1基因; 3）它们含有一个IncP质粒的oriT; 4）它们含有IncP质粒的incC1基因; 5）它们含有λ噬菌体的cos位点，cos位点位于T-DNA外部; 6）它们含有在大肠杆菌中表达的耐药基因和农杆菌属的细菌; 7）它们含有农杆菌属细菌的T-DNA右侧序列; 8）它们含有农杆菌属细菌的T-DNA左边界序列; 9）它们含有位于7）和8之间的植物转化的选择性标记基因并在植物中表达; 和10）它们含有位于7）和8之间的限制性内切核酸酶识别位点，用于克隆外来基因。

公开(公告)号：US20080301832A1

公开(公告)日：2008-12-04

申请号：US10576693

申请日：2004-10-22

申请人： Tomoaki Kubo ,  Toshihiko Komari ,  Satoru Usami ,  Yoshimitsu Takakura ,  Yukoh Hiei ,  Yuji Ishida

发明人： Tomoaki Kubo ,  Toshihiko Komari ,  Satoru Usami ,  Yoshimitsu Takakura ,  Yukoh Hiei ,  Yuji Ishida

IPC分类号： C12N15/11 ,  C12Q1/68 ,  A01H5/00 ,  C12P19/28

CPC分类号： A01H1/00 ,  A01H1/04 ,  C12N15/8201 ,  C12N15/8261 ,  Y02A40/146

摘要： The present invention provides a method for selecting genomic DNA fragments which are useful for providing a plant with an agriculturally advantageous improvement.The method of the present invention comprises the steps of: 1) preparing genomic DNA from a plant, which is then cloned into a cloning vector to form a genomic DNA library; 2) introducing a genomic fragment from each of the genomic clones constituting the genomic DNA library separately into a plant to produce transgenic plants; 3) cultivating the transgenic plants or progeny thereof to select a plant exhibiting an agriculturally advantageous phenotypic variation; and 4) selecting the genomic DNA fragment, which was introduced in step (2) into the plant selected in step (3), as a purposed genomic DNA fragment.

摘要翻译： 本发明提供了一种选择可用于提供具有农业上有利改进的植物的基因组DNA片段的方法。 本发明的方法包括以下步骤：1）从植物制备基因组DNA，然后将其克隆到克隆载体中以形成基因组DNA文库; 2）将构成基因组DNA文库的每个基因组克隆的基因组片段分别引入植物中以产生转基因植物; 3）培育转基因植物或其后代以选择表现出农业上有利的表型变异的植物; 和4）将在步骤（2）中引入的基因组DNA片段选择在步骤（3）中选择的植物中，作为目的基因组DNA片段。

公开(公告)号：US08298819B2

公开(公告)日：2012-10-30

申请号：US12306163

申请日：2007-06-25

申请人： Yoshimitsu Takakura ,  Toshihiko Komari ,  Yuji Ishida ,  Toshiyuki Komori ,  Yukoh Hiei ,  Toshiki Mine ,  Teruyuki Imayama

发明人： Yoshimitsu Takakura ,  Toshihiko Komari ,  Yuji Ishida ,  Toshiyuki Komori ,  Yukoh Hiei ,  Toshiki Mine ,  Teruyuki Imayama

IPC分类号： C12N15/82 ,  C12N15/19 ,  C12N15/00

CPC分类号： C12N15/82 ,  C12N15/8205

摘要： The present invention aims to provide novel vectors for plant transformation.The vectors of the present invention are cosmid vectors having a full length of 15 kb or less characterized in that: 1) they contain an origin of replication of an IncP plasmid, but do not contain any origin of replication of other plasmid groups; 2) they contain the trfA1 gene of an IncP plasmid; 3) they contain an oriT of an IncP plasmid; 4) they contain the incC1 gene of an IncP plasmid; 5) they contain a cos site of lambda phage and the cos site is located outside the T-DNA; 6) they contain a drug resistance gene expressed in E. coli and a bacterium of the genus Agrobacterium; 7) they contain a T-DNA right border sequence of a bacterium of the genus Agrobacterium; 8) they contain a T-DNA left border sequence of a bacterium of the genus Agrobacterium; 9) they contain a selectable marker gene for plant transformation located between 7) and 8) and expressed in a plant; and 10) they contain restriction endonuclease recognition site(s) located between 7) and 8) for cloning a foreign gene.

摘要翻译： 本发明旨在提供用于植物转化的新型载体。 本发明的载体是全长为15kb或更小的粘粒载体，其特征在于：1）它们含有IncP质粒的复制起点，但不含有其他质粒组的任何复制起点; 2）它们含有IncP质粒的trfA1基因; 3）它们含有一个IncP质粒的oriT; 4）它们含有IncP质粒的incC1基因; 5）它们含有λ噬菌体的cos位点，cos位点位于T-DNA外部; 6）它们含有在大肠杆菌中表达的耐药基因和农杆菌属的细菌; 7）它们含有农杆菌属细菌的T-DNA右侧序列; 8）它们含有农杆菌属细菌的T-DNA左边界序列; 9）它们含有位于7）和8之间的植物转化的选择性标记基因并在植物中表达; 和10）它们含有位于7）和8之间的限制性内切核酸酶识别位点，用于克隆外来基因。

公开(公告)号：US08039687B2

公开(公告)日：2011-10-18

申请号：US10576693

申请日：2004-10-22

申请人： Tomoaki Kubo ,  Toshihiko Komari ,  Satoru Usami ,  Yoshimitsu Takakura ,  Yukoh Hiei ,  Yuji Ishida

发明人： Tomoaki Kubo ,  Toshihiko Komari ,  Satoru Usami ,  Yoshimitsu Takakura ,  Yukoh Hiei ,  Yuji Ishida

IPC分类号： C12N15/82 ,  C12N15/10 ,  C12N15/29 ,  C12N15/55 ,  C12N15/70 ,  C12N15/84

CPC分类号： A01H1/00 ,  A01H1/04 ,  C12N15/8201 ,  C12N15/8261 ,  Y02A40/146

摘要： The present invention provides a method for selecting genomic DNA fragments which are useful for providing a plant with an agriculturally advantageous improvement. The method of the present invention comprises the steps of: 1) preparing genomic DNA from a plant, which is then cloned into a cloning vector to form a genomic DNA library; 2) introducing a genomic fragment from each of the genomic clones constituting the genomic DNA library separately into a plant to produce transgenic plants; 3) cultivating the transgenic plants or progeny thereof to select a plant exhibiting an agriculturally advantageous phenotypic variation; and 4) selecting the genomic DNA fragment, which was introduced in step (2) into the plant selected in step (3), as a purposed genomic DNA fragment.

摘要翻译： 本发明提供了一种选择可用于提供具有农业上有利改进的植物的基因组DNA片段的方法。 本发明的方法包括以下步骤：1）从植物制备基因组DNA，然后将其克隆到克隆载体中以形成基因组DNA文库; 2）将构成基因组DNA文库的每个基因组克隆的基因组片段分别引入植物中以产生转基因植物; 3）培育转基因植物或其后代以选择表现出农业上有利的表型变异的植物; 和4）将在步骤（2）中引入的基因组DNA片段选择在步骤（3）中选择的植物中，作为目的基因组DNA片段。

公开(公告)号：US09284567B2

公开(公告)日：2016-03-15

申请号：US13812412

申请日：2011-07-29

申请人： Yukoh Hiei ,  Yuji Ishida

发明人： Yukoh Hiei ,  Yuji Ishida

IPC分类号： A01H4/00 ,  C12N15/90 ,  C12N15/82

CPC分类号： C12N15/8205 ,  A01H4/008

摘要： An object of the present invention is to provide a method of gene introduction, which can transform a Hordeum plant at a higher efficiency compared to that in known Agrobacterium methods, and a method of producing a transformed Hordeum plant. The method of the invention includes a step of subjecting an immature embryo tissue of a Hordeum plant to centrifugation treatment and/or pressurization treatment before the inoculation with Agrobacterium, during the coculture step, and/or after the coculture step, and is characterized in that the coculture medium satisfies at least one of a) containing an antiauxin; b) containing a cytokinin; and c) containing a phenoxy auxin in an amount of less than 2 μM and/or a benzoic auxin in an amount of less than 5 μM, or not containing any phenoxy auxin and/or benzoic auxin.

摘要翻译： 本发明的目的是提供一种基因导入方法，其可以与已知的农杆菌方法相比以更高的效率转化大麦植物，以及生产转化的大麦草植物的方法。 本发明的方法包括在共培养步骤期间和/或共培养步骤之后，在土壤杆菌接种之前和/或共培养步骤之后，使大麦植物的未成熟胚胎组织进行离心处理和/或加压处理的步骤，其特征在于 共培养培养基满足以下中的至少一种：a）含有抗癌素; b）含有细胞分裂素; 和c）含有少于2μM的苯氧基生长素和/或小于5μM的苯甲酸生长素，或不含任何苯氧基生长素和/或苯甲酸生长素。

公开(公告)号：US07902426B1

公开(公告)日：2011-03-08

申请号：US10089695

申请日：2000-08-03

申请人： Yukoh Hiei ,  Keisuke Kasaoka ,  Yuji Ishida

发明人： Yukoh Hiei ,  Keisuke Kasaoka ,  Yuji Ishida

IPC分类号： C12N15/82

CPC分类号： C12N15/8201 ,  C12N15/8205 ,  C12N15/8206

摘要： A method for gene transfer by which higher efficiency for gene transfer than that by the conventional Agrobacterium method may be attained simply and without injuring the tissue is disclosed. According to the method of the present invention, the efficiency of gene transfer into plant cells by a bacterium belonging to genus Agrobacterium is promoted by accompanying heat treatment and centrifugation treatment of the plant cells or plant tissue.

摘要翻译： 公开了一种用于基因转移的方法，其通过简单地实现基因转移的效率高于常规农杆菌法，而不损伤组织。 根据本发明的方法，通过伴随对植物细胞或植物组织的热处理和离心处理，促进了属于农杆菌属的细菌的基因转移到植物细胞中的效率。

公开(公告)号：US08357836B2

公开(公告)日：2013-01-22

申请号：US12935525

申请日：2009-03-24

申请人： Yuji Ishida ,  Yukoh Hiei

发明人： Yuji Ishida ,  Yukoh Hiei

IPC分类号： A01H1/00

CPC分类号： C12N15/8205 ,  C12N15/821 ,  C12N15/8212

摘要： An Agrobacterium-mediated method for producing transformed maize or rice, culturing an Agrobacterium-inoculated immature embryo with a coculture medium and a regeneration step for culturing the immature embryo with a regeneration medium either without callus growth or after callus growth culture to regenerate whole transformed maize or rice. The method further includes a transformation enhancement and the method does not include any selection step based on the properties of a nucleic acid to be introduced by Agrobacterium in any step from coculture to regeneration.

摘要翻译： 用于生产转化的玉米或水稻的农杆菌介导的方法，用共培养培养基培养土壤杆菌接种的未成熟胚胎和用再生培养基培养未成熟胚胎的再生步骤，无需愈伤组织生长或在愈伤组织生长培养后再生整个转化的玉米 或米饭。 该方法还包括转化增强，并且该方法不包括基于在从共培养至再生的任何步骤中被农杆菌引入的核酸的性质的任何选择步骤。

公开(公告)号：US20120124696A1

公开(公告)日：2012-05-17

申请号：US13387370

申请日：2010-07-29

申请人： Yuji Ishida ,  Yukoh Hiei

发明人： Yuji Ishida ,  Yukoh Hiei

IPC分类号： A01H1/06 ,  C12N15/82

CPC分类号： C12N15/8205

摘要： The method of the present invention includes the step of excising one or more portions selected from a radicle, a germ, and an embryonic axis of a plant tissue inoculated with Agrobacterium after cultivation in a coculture medium. The present invention provides a method of gene introduction that can transform a Triticum plant at high efficiency compared to conventionally known Agrobacterium methods, and provides a method of producing a transformed plant.

摘要翻译： 本发明的方法包括在共培养培养基中培养后切除选自土壤杆菌接种的植物组织的胚根，胚芽和胚轴的一个或多个部分的步骤。 本发明提供了一种基因导入方法，其可以与常规已知的农杆菌方法相比高效地转化小麦植物，并提供了转化植物的生产方法。

公开(公告)号：US20110131685A1

公开(公告)日：2011-06-02

申请号：US12086426

申请日：2006-12-13

申请人： Yuji Ishida ,  Yukoh Hiei ,  Jun Ueki ,  Takeshi Yamamoto

发明人： Yuji Ishida ,  Yukoh Hiei ,  Jun Ueki ,  Takeshi Yamamoto

IPC分类号： C12N15/82

CPC分类号： C12N15/8205

摘要： The present invention provides a method for Agrobacterium-mediated gene transfer into a plant material, which comprises inoculating an Agrobacterium into the plant material in the presence of a powder. In the method of the present invention, the powder at least does not affect living tissues and has one or more properties selected from the group consisting of: being insoluble in water; having an affinity for living tissues; having adsorption properties; and having a surface polarity. The present invention also provides a method for producing a transformed plant, which comprises using the gene transfer method of the present invention.

摘要翻译： 本发明提供了将土壤杆菌介导的基因转移到植物材料中的方法，其包括在粉末存在下将土壤杆菌接种到植物材料中。 在本发明的方法中，所述粉末至少不影响活体组织，并且具有选自下组的一种或多种性质：不溶于水; 对活组织有亲和力; 具有吸附性能; 并具有表面极性。 本发明还提供一种生产转化植物的方法，其包括使用本发明的基因转移方法。