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Gene for increasing the production of plant biomass and/or seeds and method for use thereof

Inactive Publication Date: 2011-03-31
TOYOTA JIDOSHA KK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0015]As a result of intensive studies to achieve the above object, the present inventors have made the novel finding that the production of plant biomass can be drastically increased by causing overexpression of a gene encoding protein phosphatase 2C having characteristic consensus sequences. Thus, they have completed the present invention.
[0020]measuring the amount of biomass of a progeny plant of the transformed plant and then selecting a line in which the amount of biomass is significantly improved.
[0028]The plant according to the present invention is a plant capable of producing significantly improved amount of biomass and / or seeds compared with wild-type plants. Also, the method for increasing the production of biomass and / or seeds according to the present invention can realize the significantly increased production of biomass and / or seeds compared with target wild-type plants. Furthermore, the method for producing a plant according to the present invention makes it possible to produce a plant capable of producing significantly improved amount of biomass and / or seeds compared with wild-type plants. Therefore, through application of the present invention, for example, productivity can be improved when the plant itself is a product and this can be achieved at lower cost. Also, through application of the present invention, for example, the productivity can be improved when seeds are directly products or ingredients contained in seeds are directly products and this can be achieved at lower cost.

Problems solved by technology

However, it is thought that since many genes are involved in the amount of plant biomass (a so-called “kind of quantitative trait”), individual gene introduction or individual genetic modification cannot lead to an effective increase in production.
Meanwhile, a great deal of difficulties are associated with introduction of many genes in a desired state into a plant.
Such gene introduction is also problematic in that if successful introduction takes place, desirable traits cannot always be acquired.
However, none of these techniques can be said to exert sufficient effects of increasing the production of biomass.

Method used

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  • Gene for increasing the production of plant biomass and/or seeds and method for use thereof
  • Gene for increasing the production of plant biomass and/or seeds and method for use thereof
  • Gene for increasing the production of plant biomass and/or seeds and method for use thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0115]Preparation of Transformants (Arabidopsis thaliana) Through Introduction of the PP2C (Protein Phosphatase 2C) Gene (At3g05640)

1. Materials and Methods

1-1. Experimental Materials

[0116]As experimental materials, seeds of Arabidopsis thaliana mutants (Activation-tag T-DNA lines: Weigel T-DNS lines, Total of 20072 lines) were used. In addition, the seeds were purchased from the Nottingham Arabidopsis Stock Centre (NASC). Regarding the seeds used as experimental materials, Weigel, D. et al., 2000, Plant Physiol. 122, 1003-1013 can be referred to.

1-2. Methods

1-2-1. Selection of Salt-Resistant Mutants

[0117]Seeds of Weigel T-DNA lines were aseptically sowed on 125 mM or 150 mM NaCl-containing modified MS agar (1%) medium [vitamins in B5 medium, 10 g / l sucrose, and 8 g / L agar (for bacterial medium; Wako Pure Chemical Industries, Ltd.)] and then cultured at 22° C. under 30-100 μmol / m2 / sec illumination (a cycle of 16 hours in the light / 8 hours in the dark). Two to 4 weeks after sowing, s...

example 2

[0141]Preparation of Transformants (Rice) Through Introduction of the PP2C (Protein Phosphatase 2C) cDNA (At3g05640)

2. Materials and Methods

2-1. Experimental Materials

[0142]As experimental materials, Arabidopsis transformants into which a fragment containing ORF of the PP2C gene (At3g05640) prepared in 1 was introduced into Arabidopsis thaliana and rice (Oryza sativa L. ssp. japonica (cv. Nipponbare)) to make transformants as experimental materials.

2-2. Methods

[0143]2-2-1. Obtainment of PP2C (Protein Phosphatase 2C) cDNA (At3g05640)

[0144]Arabidopsis transformants prepared by introduction of a fragment containing ORF of the PP2C gene (At3g05640) prepared in 1 were grown until the plants reached 4 weeks of age. Total RNA was isolated from the above-ground parts, then RT-PCR was performed using TaqMan Reverse Transcription Reagents (Applied Biosystems), so that cDNA was prepared.

[0145]PCR was performed using the following primers that had been designed based on the nucleotide sequence ...

example 3

[0154]Preparation of Transformant (Rice) Through Introduction of PP2C (Protein Phosphatase 2C) cDNA (Os05g0358500)

3. Materials and Methods

3-1. Experimental Materials

[0155]As experimental materials, rice (Oryza sativa L. ssp. japonica (cv. Nipponbare)) was used.

3-2. Methods

[0156]3-2-1. Obtainment of Rice PP2C (Protein Phosphatase 2C) cDNA (Os05g0358500)

[0157]In this Example, a rice homologous gene (PP2C gene (Os05g0358500)) homologous to PP2C (protein phosphatase 2C) (At3g05640) used in Examples 1 and 2 was used. The entire sequence was chemically synthesized based on the nucleotide sequence (SEQ ID NO: 6) of the coding region of rice PP2C (Os05g0358500). A fragment of the chemically synthesized entire sequence was cloned into pDONR 221 that was a donor clone of a MultiSite Gateway Three-Fragement Vector Construction Kit (Invitro gen).

3-2-2. Construction of Plant Expression Vector

[0158]A corn-derived ubiquitin gene promoter (SEQ ID NO: 33: Christensen, A. H. and Quail, P. H., Transge...

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Abstract

A technique by which the production of plant biomass can be significantly increased is provided. A protein phosphatase 2C gene having 3 consensus sequences comprising the amino acid sequences shown in SEQ ID NOS: 1-3 from the N-terminal side in such order is over-expressed.

Description

TECHNICAL FIELD [0001]The present invention relates to: a plant in which a given gene is over-expressed; a method for increasing the production of biomass and / or seeds through overexpression of a given gene; and a method for producing such plant capable of producing an increased amount of biomass and / or seeds.BACKGROUND ART [0002]The term “biomass” generally refers to the total amount of organisms that inhabit or exist in a given area. When such term is used with regard to plants, in particular, it refers to dry weight per unit area. Biomass units are quantified in terms of mass or energy. The expression “biomass” is synonymous with “Seibutsutairyo” or “Seibutsuryo.” In the case of plant biomass, the term “standing crop” is occasionally used for “biomass.” Since plant biomass is generated by fixing atmospheric carbon dioxide with the use of solar energy, it can be regarded as so-called “carbon-neutral energy.” Accordingly, an increase of plant biomass is effective for global environ...

Claims

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Application Information

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IPC IPC(8): C12N15/87A01H5/00
CPCC12N15/8261C12N9/16Y02A40/146C12Y301/03016
Inventor KONDO, SATOSHIOHTO, CHIKARAMITSUKAWA, NORIHIROMURAMOTO, NOBUHIKOOGAWA, KENICHISUGIMOTO, HIROKITANAKA, TOMOKOYONEKURA, MADOKA
Owner TOYOTA JIDOSHA KK
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