Wheat with altered branching enzyme activity and starch and starch containing products derived therefrom

A kind of enzymatic activity, technology of wheat, applied in non-food products, wheat grain, food, starch, to obtain said crop fields

Active Publication Date: 2006-12-06
COMMONWEALTH SCI & IND RES ORG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] Therefore, wheat with an amylose content of more than 50% is not known, and although high-amylose varieties of maize and barley are known, the

Method used

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  • Wheat with altered branching enzyme activity and starch and starch containing products derived therefrom
  • Wheat with altered branching enzyme activity and starch and starch containing products derived therefrom
  • Wheat with altered branching enzyme activity and starch and starch containing products derived therefrom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0224] Embodiment 1 Materials and methods

[0225] Carbohydrate Determination and Analysis

[0226] Starch was isolated from wheat kernels using the method of Schulman et al. (1991). Starch composition was determined using the Megazyme (Bray, Co Wicklow, Republic of Ireland) kit of analytical tools and compared to control plants. Subtracting the starch weight from the total kernel weight yields the total non-starch content of the kernel to determine whether the reduced total weight is due to a reduction in the starch content.

[0227] The amylose content of the starch was determined by the colorimetric (iodine titration) method of Morrison and Laignelet (1983), slightly modified as follows. Accurately weigh approximately 2 mg (to the nearest 0.1 mg) of starch into a 2 mL screw cap test tube with a rubber gasket lid; to remove grease, add 1 mL of 85% (v / v) methanol to the starch and place the test tube in Heat in a water bath at 65°C for 1 hour, stirring the water occasional...

Embodiment 2

[0232] Example 2 Changes in genetic structure of wheat SBEIIa and SBEIIb characterization

[0233] Double-stranded RNA (Duplex-RNA, dsRNA) constructs were used to reduce the expression of wheat SBEIIa or SBEIIb genes. In such a configuration, the desired nucleic acid sequence coincides with a portion of the SBEIIa or SBEIIb gene in both the functional and antifunctional orientations relative to the promoter region, and thus the characterized RNA contains complementary regions to enable base pair to form double-stranded RNA. When part of the RNA is transcribed in transgenic plants, the intron sequences contained in the isolated region of the functional and antifunctional sequences are truncated to form tight "hairpin" doublets. The inclusion of intragenic regions was found to increase the efficiency of gene silencing due to the double RNA structure (Smith et al, 2000). The desired nucleic acid is synthesized with high molecular weight glutenin (HMWG) promoter sequence (Dx5 su...

Embodiment 3

[0237] Embodiment 3 Transformation of wheat

[0238] The genetic construct for wheat transformation was introduced by electroporation into disarmed Agrobacterium sp. LBA 4404 carrying the vir plasmids pAL4404 and pSB1, followed by media selection on spectinomycin. The engineered Agrobacterium strains were cultured in solidified YEP medium for 2 days at 27°C. The thalli were then collected and resuspended in TSIM1 with 400 mM acetosyringone and an optical density of 2.4 at 650 nm (containing 100 mg per liter of inositol, 10 g per liter of glucose, 50 mg per liter of MES buffer at pH 5.5 solution of MS medium) for wheat inoculation.

[0239] Wheat species (variety NB1, a spring wheat variety supplied by Nickerson Seeds Ltd, Rothwell, Lincs.) were grown in a greenhouse with a day / night temperature of 22 / 15°C and 16 hours of light per day. Young shoots (embryo approximately 1 mm long) were harvested 14 days after anthesis along with 50 cm young shoot stems. Remove all but the f...

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Abstract

Wheat having a reduced level of SBEIIa activity, that may have a relative high amylose content. Wheat having a mutant SBEIIa gene in the A genome. The wheat might additionally have reduced levels of SBEIIb activity. The wheat grain of this invention can be of a non-shrunken phenotype despite a lesion in the amylopectin synthesis pathway, and may also have a high relative amylose content.

Description

technical field [0001] The present invention relates to a wheat crop having a relatively high amylose content in the kernels. The invention also relates to a wheat plant with reduced starch branching enzyme IIa (SBEIIa) activity in the endosperm, and a method for obtaining said plant. The invention also relates to grains, starches, food and non-food products obtained therefrom. Background technique [0002] In cereals, starch accounts for about 45% to 56% of the weight of mature grains. [0003] Starch contains two types of molecules: amylose and branched starch. Amylose is an essentially linear molecule consisting of oc-1.4 linked to glucose chains. Whereas amylopectin is highly branched and connected to a-1,6 glucosidic bonds in linear chains. [0004] In higher plants, starch synthesis in the endosperm can be synthesized by a series of biochemical enzymes that catalyze four key steps. In the first step, ADP-glucose pyrophosphorylase activates starch monomer precursor...

Claims

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Application Information

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IPC IPC(8): C12N15/52C12N15/82C12N9/10A01H1/00A01H5/00A23L1/0522A23L29/212
Inventor 阿麦德·里贾纳萨德奎·拉曼马太·肯尼迪·莫维尔中易·李
Owner COMMONWEALTH SCI & IND RES ORG
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