Plants with modified starch metabolism

A crop, amylase technology, applied in the fields of plant peptides, food science, plant products, etc., can solve the problem of the lag of wheat transformation efficiency

Inactive Publication Date: 2011-02-09
COMMONWEALTH SCI & IND RES ORG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Another reason is that the transformation efficiency of wheat lags behind that of other cereals

Method used

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  • Plants with modified starch metabolism
  • Plants with modified starch metabolism
  • Plants with modified starch metabolism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0263] Embodiment 1 Materials and methods

[0264] Extraction of starch from cereals

[0265] The mature grains were pulverized with a small pulverizer of Metefem (prototype), and the pulverized material was passed through a 0.5 mm sieve. The semolina obtained was weighed and 50% by weight of water was added to soften the result. The dough is obtained by mechanically mixing water and flour. Excess water was added to extract starch from gluten, and then the starch suspension was filtered through 100 μm filter paper to remove residue. Repeat the extraction 4 times or until no starch remains in the dough. The starch was granulated by centrifugation (5,000 rpm, 10 minutes, 4°C). The covering albumen is removed and the starch is resuspended in excess water. Repeat the wash 3 times. The extracted starch was freeze-dried in a FTS freeze dryer (Model No.FD-3-55D-MP). Using this method, 10 g of grain yields approximately 6 g of flour and 3 g of starch.

[0266] Extraction of st...

Embodiment 2

[0292] Identification of GWD and PWD Genes in Example 2 Wheat and Other Cereals

[0293] The amino acid sequence (Accession No. AAK11735) used for the potato R1 protein sequence and the Arabidopsis PWD protein (NP_194176) were used as query sequences in the NCBI database, and the wheat EST sequence was queried using the BLASTN program. Seventeen ESTs (listed below) were identified and aligned to form a sequence of 2273 base pairs (SEQ ID NO: 1).

[0294] dbj|CJ626658.1|CJ626658 Y.Ogihara unpublished cDNA library.

[0295] gb|CV773056.1|FGAS067452 Triticum aestivum FGAS library.

[0296] dbj|CJ694861.1|CJ694861 Y.Ogihara unpublished cDNA library.

[0297] gb|CA743865.1|wrils.pk006.k23 wrils Triticum aestivum cDNA.

[0298] gb|BQ240991.1|TaE05010D07R Ta05 Triticum aestivum cDNA.

[0299] dbj|CJ696711.1|CJ696711 Y.Ogihara unpublished cDNA library.

[0300] dbj|CJ730334.1|CJ730334 Y.Ogihara unpublished cDNA library.

[0301] gb|BQ237936.1|Ta05010D07F TaE05 Triticum aestivum ...

Embodiment 3

[0317] Example 3 Production of Transformed Plants Containing Structures That Suppress GWD Gene Expression

[0318] To examine the effects of inhibiting GWD gene expression, gene constructs capable of expressing double-stranded RNA molecules used to inhibit partial homologs of wheat GWD were designed, targeting a conserved region consistent with the starch-binding domain.

[0319] This construct contains the wheat Bx17 HMW glutenin gene promoter for expression of dsRNA, which acts as a tissue-specific promoter, preferably expressed in the cereal endosperm. Therefore, it is predicted that inhibition of gene expression may occur primarily in the endosperm.

[0320] The suppressor gene construct was assembled in the following pBx17IRcasNOT cloning vector. This vector contains the following components in order: from the wheat HMWG Bx17 gene, as reported by Reddy et al. (1993), including the endosperm-specific promoter of the first 1897 nucleotides of the Bx17 genome sequence, its ...

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Abstract

The specification provides methods of obtaining a genetically modified plant which has increased production potential compared to a control plant, the method comprising the steps of i) obtaining a plurality of plants at least one of which comprises in its genome a heterologous polynucleotide, ii) identifying from the plurality of plants a plant which has increased production potential relative to the control plant and comprises the heterologous polynucleotide, and iii) selecting the genetically modified plant, wherein the polynucleotide comprises a transcriptional control sequence operably linked to a nucleic acid sequence which encodes an agent that modifies endogenous starch phosphorylation and / or starch degradation in the plant. In some embodiments, the plant has increased endogenous glycosylase or increased digestibility compared to a control plant. In some specific embodiments, the endogenous starch phosphorylation and / or starch degradation is modified by modifying expression or activity of one or more enzymes selected from the group consisting of alpha-amylase (EC 3.2.1.1), beta-amylase (EC 3.2.1.2), glucoamylase (EC 3.2.1.3), starch phosphorylase (EC2.4.1.1), glycosylase (EC 3.1.33), sucrase-isomaltase (EC 3.2.10), amylomaltase (EC 2.4.1.25), maltase (EC 3.2.1.20), isoamylase, and a-glucan, water dikinase (GWD, EC 2.7.9.4).

Description

technical field [0001] The present invention describes methods for increasing the production potential of organisms. In particular, the present invention relates to crop starch metabolic pathways and provides crops with modified starch metabolic pathways and production potential, including gramineous crops such as wheat and barley. The present invention describes various methods of propagating crop plants with improved production potential, such as increased yield, growth, biomass, vigor, etc., and methods of producing products of interest from such modified crop plants. Background technique [0002] A detailed bibliography of publications referenced by the authors in this application is included at the end of the specification. [0003] Any prior publications (or information derived from such prior publications) or any known event to which the present invention refers, prior publications (or information derived from such prior publications) that are common general knowledg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N9/00C12N15/29A01H5/00C12N9/12A23L7/10A23L7/104A23L7/109
CPCC07K14/415C12N15/8245C12Y207/09004C12N9/1294C12N15/8234C12N15/8261
Inventor 吉恩-菲利普·弗朗克斯·米歇尔·拉尔钟仪·李马修·肯尼迪·莫瑞尔
Owner COMMONWEALTH SCI & IND RES ORG
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