Method for generating frangrance flavor through inactivation or reduction functional protein with betaine aldehyde dehydrogenase (BADH) activity

A protein and functional technology, applied in biochemical equipment and methods, redox enzymes, recombinant DNA technology, etc., can solve problems such as limited knowledge of biochemistry and genetics, molecular genetics that have not been described, and genes that have not been identified

Inactive Publication Date: 2007-10-24
GRAIN FOODS CRC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the biological mechanisms leading to the development of the aroma remain unknown
Although fragrance is associated with the production of volatile compounds, such as 2-acetyl-1-pyrroline, the biological processes leading to the production and attenuation of these volatile compounds remain unknown
[0006] Furthermore, the molecular genetics of aroma production in any organism has not been described, and prior to the present invention no genes associated with this trait had been identified
[0007] Existing biochemical and genetic knowledge related to scent production is limited, and it has not previously been possible to manipulate scent-free organisms to produce scent, or to efficiently identify scent-producing, or at least capable, scent-producing organisms

Method used

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  • Method for generating frangrance flavor through inactivation or reduction functional protein with betaine aldehyde dehydrogenase (BADH) activity
  • Method for generating frangrance flavor through inactivation or reduction functional protein with betaine aldehyde dehydrogenase (BADH) activity
  • Method for generating frangrance flavor through inactivation or reduction functional protein with betaine aldehyde dehydrogenase (BADH) activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0280] introduction

[0281] To identify the chromosomal location of genes encoding the rice aroma phenotype, simple sequence repeats (SSR) or microsatellite 2 and single nucleotide polymorphisms (SNPs) 3 Markers are mapped against segregating populations. Rice Genome Sequence 14 The availability of allows comparison of sequences of scented and non-scented genotypes. This allowed targeted resequencing of genes and genome sequences in the aroma genotype to the most probable portion of chromosome 8.

[0282] Materials and methods

[0283] plant material

[0284] In this study, 168 wild-grown F 2 Populations of individuals derived from Kyeema (Pelde / / Della / Kulu) (tall, jasmine-scented, scented, long-grain, Australian cultivar) and Gulfmont (Lebonnet / / CI9881 / PI 331581) (early maturing, semi-dwarf, non-scented, long-grain, American cultivar).

[0285] Following genetic mapping, candidate genes were identified in the region between the flanking markers of the published geno...

Embodiment 2

[0311] For any single analyst whose senses would become saturated, or often physically injured by the abrasion of the tongue from chewing hard grains, the ability to distinguish between scented and non-scented samples would increase with each successive analysis. decrease as the analysis proceeds.

[0312] Described below is the development of a PCR assay to genotype rice aroma.

[0313] plant material

[0314] All rice samples were provided by Yanco Agricultural Institute, NSW Agriculture. In addition to 168 wild-grown F 2 Individual populations were also validated using different collections of 14 scented and 74 non-scented varieties, where the F 2 Individuals derived from Kyeema (Pelde / / Della / Kulu) (tall, jasmine-scented, long-grained, Australian cultivar) and Gulfmont (Lebonnet / / CI9881 / PI 331581) (early, semi-dwarf, non-scented , long-grain, American cultivars).

[0315] genetic mapping

[0316] Genetic mapping was performed as described in Example 1.

[0317] Prime...

Embodiment 3

[0355] Materials and methods for generating transgenic wheat with BAD2 RNAi

[0356] Design RNAi inserts

[0357] The RNAi insert can be derived from the 5' end of the wheat BAD2 cDNA. This region shares 76.8% homology with the same region in the BAD1 gene homolog. In this example, RNAi inserts were designed such that transgenic plants exhibited specific BAD2 interference, but not BAD1 homolog interference (Figure 28).

[0358] PCR, genotyping and sequence analysis

[0359] Oligonucleotide primers are synthesized by methods known in the art. Use 0.2 μL Platinum(R) Taq DNA polymerase (Gibco BRL(R), 2 μL cDNA, 2.5 μL 10× buffer (Gibco BRL(R), 1 μL 50 mM MgCl 2 (Gibco BRL(R)), 1 μL of dNTPs (5 mM), 2.5 μL of each primer pair - (Table 1) [2 mM], in a total volume of 25 μL for PCR. PCR was performed using Perkin Elmer, Gene Amp PCR system9700. The cycle conditions are as follows: first denaturation at 94°C for 2 minutes, followed by 30 cycles at 94°C for 30 seconds, 58°C for ...

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Abstract

The invention relates to methods of increasing production of fragrance by an organism capable of expressing a functional protein having an amino acid sequence that is at least 30% identical to the amino acid sequence of SEQ ID NO: 1, comprising reducing or eliminating the activity of the functional protein in the organism. The invention also relates to methods of establishing whether an organism is capable of producing a fragrance, and to methods of producing organisms which produce fragrance.

Description

technical field [0001] The present invention relates to methods of increasing aroma production in organisms, methods of identifying aroma-producing organisms, and aroma-producing organisms. Background technique [0002] It should be understood that while prior art publications are incorporated herein by reference, it is not an admission that said publications form part of the common general knowledge in the art, in Australia or any other country. [0003] Aroma is an attractive feature of many foods. For example, the aroma associated with rice varieties such as Basmati 370, KhaoDawk Mali 105, Kyeema, Dumsorhk, Dellmont, Amber and Goolarah rice varieties makes these particular rice varieties very popular as a valuable food source. Fragrance is also associated with other cereal species such as oats, barley, wheat and non-cereal plants such as Pandanus amaryllifolius leaves. However, many plants and other organisms do not produce fragrance, or produce only very small amounts ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N5/14C12N15/09
CPCC12N15/8243C12N9/0008
Inventor 罗伯特·J·亨利金庆生丹尼尔·L·E·沃特斯路易斯·M·布拉德伯里蒂莫西·L·菲茨杰拉尔德
Owner GRAIN FOODS CRC
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