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Method of producing plastid transformants

A technology of transformants and plastids, applied in biochemical equipment and methods, chemical instruments and methods, using vectors to introduce foreign genetic materials, etc., can solve the problems of increasing the number of processes, not knowing the applicability of monocotyledonous plants, etc., and achieve excellent results The effect of screening efficiency

Inactive Publication Date: 2017-08-29
HONDA MOTOR CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in this method, it is necessary to prepare a strain that properly forms a greening callus by performing transformation of the nuclear genome in advance, and the applicability to other monocotyledonous plants is not known except for increasing the number of processes

Method used

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  • Method of producing plastid transformants
  • Method of producing plastid transformants
  • Method of producing plastid transformants

Examples

Experimental program
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Effect test

Embodiment 1

[0103] Preparation of switchgrass regenerated strain

[0104] Mature seeds of switchgrass variety "Alamo" were dehusked by 60% (v / v) sulfate treatment, and seed fungicides were used for sterilization. The sterilized seeds were further surface sterilized by hypochlorous acid treatment and kept in callus induction medium (PVC medium: MS salts and vitamins, 30 g / L maltose, 22.6 μM 2,4-dichlorophenoxy Glycolic acid, 4.4μM BAP, 2.5mM MES, 3g / L gellan gum, pH 5.7). Seeds were grown at 28°C in the dark for several months. Then, the growing calli were sorted into independent strains for each derived seed. Strains in which compact callus growth expected to have regenerative capacity are selected and cultured. In addition, the compact calli of selected strains were kept in regeneration medium (PVS medium: MS salts and vitamins, 30 g / L maltose, 1.4 μM gibberellin A 3 , 2.5mM MES, 3g / L gellan gum, pH 5.7), cultured at 28°C under light for 4 weeks, and then confirmed the regeneration ...

Embodiment 2

[0135] Introduction of genes into rice plastids and selection of transformed cells

[0136] The PSR1 gene was introduced into the plastids of rice variety Nipponbare and rice variety Koshihikari, and transformants were selected using sodium nitrite. Specifically, proceed as follows. MS-NK medium (Nishimura et al., Nature Protocols, 2006, Vol.1, p. 2796-2802) used as a regeneration medium initially contained nitrate nitrogen. The wild-type rice variety Nipponbare has regeneration ability in MS-NK medium containing sodium nitrite at a concentration of 400 mg / L. However, wild-type rice variety Koshihikari did not regenerate even in MS-NK medium with weak nitrite reductase and no addition of sodium nitrite.

[0137] Calli from blastocysts cultured from mature rice seeds were induced and prepared using methods in the related art. The obtained calli of about 2 mm in diameter were arranged in a circle of about 35 mm in diameter in the central part of N6D medium (Nishimura et al., ...

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Abstract

A method of producing plastid transformants including a process (a) in which genes coding for proteins having a function of neutralizing nitrite toxicity are introduced into plant tissues and a process (b) in which the plant tissues into which the genes are inserted in the process (a) are cultured in a medium containing nitrite nitrogen having a concentration at which growth of wild type plant cells is suppressed or a concentration at which regeneration of wild type plants is suppressed and thus plastid transformants obtained by inserting the genes into plastid genomes are screened.

Description

technical field [0001] The present invention relates to a method for producing a plastid transformant, more particularly, to a method for producing a plastid transformant, wherein an exogenous gene is introduced into a plant tissue having differentiated plastids, and a gene encoding a neutralizing nitrous acid Genes for salt-toxic proteins such as nitrite reductase genes are used as selectable marker genes to select transformants. [0002] This application claims the benefit of Japanese Patent Application No. 2015-169817 filed on August 28, 2015 and Japanese Patent Application No. 2016-054448 filed on March 17, 2016, the contents of which are incorporated herein by reference. Background technique [0003] Plastids (chromogens) of plants are pigment-containing organelles in plant cells that have their own genome and are self-growing. Chloroplasts, which are a type of plastid, are the most prominent feature of plant cells, performing photosynthesis, carbon, nitrogen, and sulf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
CPCC12N15/8209C12N15/821C12N15/8214C07K14/415C12N15/8241
Inventor 山口明日香大熊二郎近藤康弘广濑佳嗣
Owner HONDA MOTOR CO LTD
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