Novel isoamylases and associated methods and products
First Claim
1. A method of identifying a polyploid plant which has at least one impaired isoamylase gene, the method comprising screening the plant for a polymorphism or mutation in an isoamylase gene which is linked to reduced production of a functional isoamylase protein, wherein wildtype plants accumulate starch in a storage organ of said wildtype plants, and wherein the corresponding isoamylase gene of said wildtype plants produce functional isoamylase in said storage organ.
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Abstract
The present invention provides methods for distinguishing between different isoamylase genes of individual genomes of a polyploid plant. These methods enable a polyploid plant-to be produced and/or identified which has at least one impaired isoamylase gene. In particular, the methods of the invention enable various plants of a polyploidy species to be produced and/or indentified which, between the plants, lack a functional isoamylase gene for each genome of the plant. Such plants can be cross-bred to produce a plant which lacks any functional isoamylase protein. Furthermore, the invention provides numerous isoamylase gene, protein and cDNA sequences from various polyploidy plant species, including hexaploid wheat.
10 Citations
58 Claims
- 1. A method of identifying a polyploid plant which has at least one impaired isoamylase gene, the method comprising screening the plant for a polymorphism or mutation in an isoamylase gene which is linked to reduced production of a functional isoamylase protein, wherein wildtype plants accumulate starch in a storage organ of said wildtype plants, and wherein the corresponding isoamylase gene of said wildtype plants produce functional isoamylase in said storage organ.
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18. A method of producing and identifying a polyploid plant which has at least one impaired isoamylase gene, the method comprising;
- i) exposing a polyploid plant, or a progenitor thereof, to conditions which promote DNA mutagenesis, and ii) screening the plants obtained from step i), or progeny thereof, to identify plants which have at least one impaired isoamylase gene.
- View Dependent Claims (19, 33, 35, 36, 37, 38, 39, 40, 41, 42)
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22. A method of producing and identifying a polyploid plant which has at least two impaired isoamylase genes, the method comprising;
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i) exposing a first polyploid plant, or a progenitor thereof, to conditions which promote DNA mutagenesis, ii) screening the plants obtained from step i), or progeny thereof, to identify plants which have at least one impaired isoamylase gene, iii) crossing the plant of step ii) which has at least one impaired isoamylase gene with a second polyploid plant which has at least one impaired isoamylase gene, wherein the first and second plants have at least one impaired isoamylase gene which is on a different genome when compared to the other plant, and iv) screening the plants obtained from step iii), or progeny thereof, to identify plants which have at least two impaired isoamylase genes. - View Dependent Claims (23, 24)
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25. A method of producing a polyploid plant which has reduced levels of functional isoamylase protein, the method comprising;
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i) crossing two parent polyploid plants each of which have at least one impaired isoamylase gene, and ii) screening the plants obtained from step i), or progeny thereof, to identify plants which have at least two impaired isoamylase genes, wherein each of the parent plants have at least one impaired isoamylase gene which is on a different genome when compared to the other parent plant. - View Dependent Claims (26, 27, 28)
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29. A method of introducing an impaired isoamylase gene into the genome of a polyploid plant, the method comprising;
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i) crossing a first parent plant with a second parent plant, wherein the as second plant comprises at least one impaired isoamylase gene, and ii) backcrossing the progeny of i) with plants of the same genotype as the first parent plant for a sufficient number of times to produce a plant with a majority of the genotype of the first parent but comprising the impaired isoamylase gene, wherein progeny plants are screened for the impaired isoamylase gene. - View Dependent Claims (30, 43)
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- 31. A method of distinguishing between the A and B genomes of wheat, the method comprising detecting a nucleotide sequence difference between the isoamylase gene of the A and B genomes, wherein the plant expresses isoamylase in the endosperm of developing grain.
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34. A non-transgenic polyploid plant comprising at least two impaired isoamylase genes, wherein wildtype plants of the same species of the polyploid plant accumulate starch in a storage organ of said wildtype plants, and wherein the corresponding isoamylase genes of said wildtype plants produce functional isoamylase in said storage organ.
- 44. An antibody that binds to at least one isoform of an isoamylase from a polyploid plant.
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47. A substantially purified polypeptide comprising an amino acid sequence selected from the group consisting of:
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i) a sequence provided as SEQ ID NO;
20,ii) a sequence provided as SEQ ID NO;
21,iii) a sequence provided as SEQ ID NO;
22,iv) a sequence that is at least 93% identical to any one of i) to iii) which is at least 535 amino acids in length, v) a sequence provided as SEQ ID NO;
23,vi) a sequence provided as SEQ ID NO;
24,vii) a sequence provided as SEQ ID NO;
25, andviii) a sequence which is at least 93% identical to any one of v) to vii). - View Dependent Claims (48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58)
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Specification