Method of elevating GGT activity of plant, plant with elevated GGT activity and method of constructing the same

A plant, a technology of transgenic plants, applied to one or more of the group, the new utilization of GGT and its encoded gene, the field of gene utilization, can solve the problems of unreported disclosure, unknown genes, etc.

Inactive Publication Date: 2009-01-28
AJINOMOTO CO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, the glyoxylate aminotransferase gene that plays a role in peroxidase has not been identified, but recently Liepman et al. Alanine in oxisomes: glutamate-glyoxylate aminotransferase (Plant. J. 25:487-498), but the gene for glutamate-glyoxylate aminotransferase remains unknown
Moreover, it is not necessarily clear how this glutamic acid glyoxylate aminotransferase activity affects the various amino acid content, the increase or decrease of the total amino acid content, photosynthetic energy, and stress tolerance including glutamic acid in plants. There is no report on the role of plant traits, and no reports reveal the possibility of improving plant traits, especially the actual improvement in plant possibility of total amino acid content and / or specific amino acid content in body or seed

Method used

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  • Method of elevating GGT activity of plant, plant with elevated GGT activity and method of constructing the same
  • Method of elevating GGT activity of plant, plant with elevated GGT activity and method of constructing the same
  • Method of elevating GGT activity of plant, plant with elevated GGT activity and method of constructing the same

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Experimental program
Comparison scheme
Effect test

Embodiment

[0084] Cultivation of all plants was carried out under the following conditions:

[0085] The following components were used on the plate as the minimal medium for culturing: PNS ( Mol. Gen. Genet. 204: 430-434) or MS (Physiol Plant 15: 473-479) inorganic salts and 0.8 (w / v) agar. If cultivation is carried out on rock wool, only PNS inorganic salts are used as a nutrient source.

[0086] In addition, the obtained Arabidopsis thaliana GGT deletion strain was used as a model plant for transformation experiments. The methods for obtaining GGT-deleted strains are shown in Reference Examples 1 and 2.

reference example 1

[0087] Reference Example 1. Obtaining a GGT-deleted strain of Arabidopsis thaliana

[0088] (1) Preparation of primers for screening GGT deletion strains

[0089] The GGT gene, since it is also the AlaAT gene, was obtained from the alanine aminotransferase (AlaAT) gene information of Arabidopsis thaliana.

[0090] According to the AlaAT gene data (GenBank) published on the Internet, the copy number and sequence of AlaAT (GGT) were predicted to prepare primers. A data search was performed using alanine aminotransferase and Arabidopsis as keywords, and it was found that there were at least 4 copies of a gene that was predicted to be a GGT gene in the genome. The GenBank accession numbers of each gene are AC005292 (F26F24.16), AC011663 (F5A18.24), AC016529 (T10D10.20), AC026479 (T13M22.3). Each gene is named GGT1, GGT2, GGT3, and GGT4, respectively, and the nucleotide sequences of each cDNA are recorded as sequence numbers 1, 3, 5, and 7, and the predicted amino acid sequence...

reference example 2

[0107] Reference Example 2. Obtaining a GGT Deletion Homozygous Strain

[0108] (1) Screening of homozygotes

[0109] T2 seeds of the line with the confirmed insertion marker were sown on MS medium containing 10 mg / l hygromycin. Transplanted onto asbestos 3 weeks later DNA was extracted from approximately 5 mm square samples of Rossetto leaves. The extraction method used Li's method (Plant J.8: 457-463). To confirm homozygosity, PCR was performed using primers (AAT1U / AAT1L2) sandwiching the marker. PCR was carried out 30 cycles under the conditions of denaturation at 94°C for 30 seconds, annealing at 57°C for 30 seconds, and extension at 72°C for 60 seconds. As a control, wild-type genomic DNA was used as a template. Part of the PCR products were separated by 1% agarose gel electrophoresis. There were 11 homozygous lines among the 35 lines.

[0110] (2) Detection of GGT expression

[0111] The progeny of the obtained homozygous germlines were used to confirm whether g...

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Abstract

The present invention describes a novel plant having increased activity of glutamate glyoxylate aminotransferase (GGT), a method of producing the plant, a seed of the plant, and a plant having an increased amino acid content. The present invention more specifically provides a plant which is improved in the content of at least one of the amino acid selected from the group consisting of serine, arginine, glutamine, and asparagine, and a method of producing the plant, as well as the seed of the plant. A plant having increased GGT activity is produced by mutagenesis, introduction of a nucleic acid molecule and the like. A genetic construction being capable of enhancing the expression of a GGT gene, particularly a genetic construct being capable of expressing a GGT gene and/or increasing the level of an endogenous gene having GGT activity is introduced into a plant.

Description

[0001] This application is a divisional application. The filing date of the original application is August 5, 2003. The application number is 03824044.0 (PCT / JP2003 / 009946). Preparation". Background of the invention [0002] The present invention relates to a plant with increased glutamate-glyoxylate aminotransferase (GGT) activity. [0003] The present invention also relates to a method for utilizing glutamate-glyoxylate aminotransferase (GGT) and / or the gene encoding GGT. [0004] In addition, the present invention relates to increasing the amino acid content in plants and / or their seeds, especially increasing the amino acid content selected from the group consisting of serine (Ser), arginine (Arg), glutamine (Gln), asparagine (Asn) The method of one or more than one amino acid content, and the amino acid content, especially selected from the group consisting of serine (Ser), arginine (Arg), glutamine (Gln), asparagine (Asn) A plant with increased content of one or more am...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/82A01H1/00A01H5/00A23K1/14C12P13/06C12P13/10C12P13/14C12P13/20A23K10/30
Inventor 五十岚大亮大住千荣子
Owner AJINOMOTO CO INC
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