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Monocdyledon plant cells and plants which synthesise modified starch

A technology of monocotyledonous plants and plant cells, which is used in the production of this starch, food and baked goods, and can solve the problems of expensive and time-consuming chemical modification

Inactive Publication Date: 2004-10-27
BASF AGRICULTURAL SOLUTIONS SEED LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chemical modification is usually expensive and time-consuming, and the physicochemical properties of the resulting starch may differ from those modified in vivo

Method used

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  • Monocdyledon plant cells and plants which synthesise modified starch
  • Monocdyledon plant cells and plants which synthesise modified starch
  • Monocdyledon plant cells and plants which synthesise modified starch

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0245] Preparation of vector pUbiR1 for transformation of wheat plants

[0246] To prepare the vector pUbiR1, first prepare the vector pUbi.cas as follows:

[0247] Vector pUbi2afp (see Figure 2) was partially digested with restriction enzymes PstI / EcoRI. The resulting 4.19 Kb fragment contained the pUC19 vector (Yannish-Perron et al., 1985, Gene 33:103-119), the 1.5 Kb ubiquitin promoter and the first exon of the ubi gene and the shortened ubiquitin 1 Intron (ClaI deletion) (Christensen et al., Plant Mol. Biol. 18, (1992), 675-689), used after gel elution.

[0248] The nos terminator was isolated from the vector pAct.cas (product number CambiaTG0063, Cambia, GPO Box 3200, Canberra ACT 2601, Australia) as a PstI / EcoRI fragment, and the two fragments were ligated to generate the vector pUbi.cas.

[0249] Subsequently, the cDNA (SEQ ID No.1) of the potato R1 gene was isolated as a partial digest (SmaI / Asp718 fragment) from the vector pRL2 (WO97 / 11188, Example 4, deposited in t...

Embodiment 2

[0251] Production of wheat plants expressing the potato R1 gene, and analysis of the phosphate content of starch from these plants

[0252] Transformation of wheat was carried out using biolistic transformation (Becker et al., Plant J. 5(2), (1994), 229-307). Vector pUbiR1 and vector p35SacS for biolistic co-transformation (see Figure 4 ) or pAct1-Fneo / CaIgus (Müller et al., Plant Science 114, (1996), 71-82, see Figure 5 ) were used in the same molar ratio in the DNA pellet precipitation batches.

[0253] The vector p35SAcS was prepared as follows: the pat gene of Streptomyces viridans (Wohlleben et al., Gene 70, (1988), 25-37) was amplified by polymerase chain reaction. The primers used were designed to create BamHI restriction sites on both sides of the amplicon. The BamHI fragment was then cloned into the BamHI restriction site placed between the 35S promoter and 35S terminator of vector pDH51 (Pietzrak et al., Nucleic Acids Res. 14, (1986), 5857-5868). The cassette c...

Embodiment 3

[0273] RVA Analysis of Plant Starch from Transgenic Wheat Plants Expressing the Potato R1 Gene, and Study of Gel Strength

[0274] Starches from the wheat plants described in Examples 1 and 2 were subjected to RVA analysis.

[0275] The results of the RVA analysis (experimental: see Methods) showed that the sticky behavior of starch from transgenic wheat plants expressing the potato R1 gene was significantly altered compared to starch from corresponding wild-type wheat plants (Florida variety) that were not genetically modified (see Table 2).

[0276] Table 2:

[0277] Series number RVA RVA RVA RVA RVA

[0278] Max Min Fin Set T

[0279](%) (%) (%) (%) (%)

[0280] Wild type (Florida) 100 100 100 100 100

[0281] 19(T0) 98.7 113.8 116.8 120.7 102

[0282] 20-25(T1) 155.3 190 193.8 198.8 99

[0283] 37(T0) 143.8 156.2 148.8 139 101

[0284] 40-11-8(T2) 156.2 185.5 187.7 190.6 97

[0285] legend:

[0286] RVA=Rapid Visco Analyzer

[0287] Max = see ...

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Abstract

The present invention relates to monocotyledon plant ells and plants which are genetically modified, wherein the genetic modification consists of the introduction of an extraneous nucleic acid molecule which codes for a protein with the biological activity of an R1 protein. The present invention further relates to means and methods for the production thereof. Plant cells and plants of this type synthesise a modified starch, which is characterised in that is has an increased phosphate content and / or a modified phosphorylation pattern and / or an increased final viscosity in an RVA profile and / or a reduced peak temperature in DSC analysis and / or an increased gel strength in the texture analysis compared with starch from corresponding non-genetically modified monocotyledon plants. Therefore, the present invention also relates to the starch which is synthesised from the plant cells and plants according to the invention, and to methods of producing said starch. The present invention further relates to wheat flours which contain said modified starches, and to food products and bakery products which contain said wheat flours and / or starch.

Description

field of invention [0001] The present invention relates to genetically modified monocotyledonous plant cells and plants, wherein the genetic modification includes introducing an exogenous nucleic acid molecule encoding a protein having the biological activity of R1 protein. The invention also relates to means and methods for their production. Such plant cells and plants synthesize a modified starch characterized by an increased phosphate content, and / or an altered phosphorylation pattern, and / or in Increased final viscosity in RVA plot, and / or decreased peak temperature in DSC analysis, and / or increased gel strength in structural analysis. Accordingly, the present invention also relates to starch synthesized by plant cells and plants according to the invention, and methods for producing such starch. The invention also relates to wheat flour containing such modified starch, and food and bakery products containing such wheat flour and / or starch. Background of the invention ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H5/00A21D2/18A21D6/00A23L1/0522A23L7/10C07K14/415C08B30/00C12N5/10C12N9/00C12N15/09C12N15/29C12N15/54C12N15/82
CPCC08B30/00A23L1/1041A23L1/0522A23V2002/00C12N15/8245C07K14/415C12N9/00A21D2/186A23L29/212A23L7/198A23V2300/21
Inventor G·舍韦P·克尼斯S·F·阿玛蒂H·洛茨D·贝克尔V·兰德许茨J·皮林
Owner BASF AGRICULTURAL SOLUTIONS SEED LLC
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