University of Florida Research Foundation, Incorporated

Research Corporation Technologies, Inc.

L Curtis Hannah

Thomas W Greene

Nikolaos Georgelis

Michael Giroux

Robert J Ferl

Vimla Vasil

Indra K Vasil

Hwa Jee Chung

Joanna Marie-France Cross

Ke Wu

Carla R Lyerly Linebarger

Brian Burger

Tom W Greene

Paul C Sehnke

Maureen A Clancy

The subject invention concerns materials and methods for providing plants or plant tissue with increased resistance to heat conditions. Increased resistance of a plant or plant tissue to heat conditions provides for decreased yield losses generally observed at elevated temperatures. One aspect of the invention concerns polynucleotides that encode a mutant plant small subunit of AGPase. The subject invention also comprises a mutant plant small subunit of AGPase encoded by a polynucleotide of the invention. The subject invention also concerns plants comprising a polynucleotide of the invention and method for making the plants.

Heat resistant plants and plant tissues and methods and materials for making and using same

The subject invention pertains to novel mutant polynucleotide molecules that encode enzymes that have increased heat stability. These polynucleotides, when expressed in plants, result in increased yield in plants grown under conditions of heat stress. The polynucleotide molecules of the subject invention encode maize endosperm ADP glucose pyrophosphorylase (AGP) and soluble starch synthase (SSS) enzyme activities. Plants and plant tissue bred to contain, or transformed with, the mutant polynucleotides, and expressing the polypeptides encoded by the polynucleotides, are also contemplated by the present invention. The subject invention also concerns methods for isolating polynucleotides and polypeptides contemplated within the scope of the invention. Methods for increasing yield in plants grown under conditions of heat stress are also provided.

Heat stable mutants of starch biosynthesis enzymes

The subject invention concerns polynucleotides encoding a small subunit of plant AGP having one or more mutations in the amino acid sequence wherein the mutation confers increased heat stability to the expressed AGP enzyme. Mutations in the N-terminus of the small subunit of heat labile plant AGP results in AGP enzymes that are significantly more heat stable compared to wild type AGP in that the mutant AGP retains significant levels of enzymatic activity following exposure to heat treatment. In one embodiment, the polynucleotide encodes a mutant small subunit of maize AGP. The subject invention also concerns methods for providing a plant with increased resistance to heat conditions. Plants with heat labile AGP can be transformed with a polynucleotide of the present invention. The subject invention also concerns these transformed plants and transgenic progeny thereof. The subject invention also concerns mutant polypeptides encoded by polynucleotides of the present invention.

Heat stable variants of plant adenosine diphosphate glucose pyrophosphorylase small subunit

The subject invention concerns materials and methods for providing plants or plant tissue with increased resistance to heat conditions and/or increased starch biosynthesis. Increased resistance of a plant or plant tissue to heat conditions provides for decreased yield losses as compared to the yield losses generally observed at elevated temperatures. One aspect of the invention concerns polynucleotides that encode a mutant plant small subunit of AGPase. The subject invention also comprises a mutant plant small subunit of AGPase encoded by a polynucleotide of the invention. The subject invention also concerns plants comprising a polynucleotide of the invention and method for making the plants.

Heat resistant plants and plant tissues comprising a variant adenosine diphosphate glucose pyrophosphorylase small subunit protein and methods of use thereof

The subject invention concerns materials and methods for providing plants or plant tissue with increased starch biosynthesis. Increased starch biosynthesis provides for increased yield. One aspect of the invention concerns polynucleotides that encode a mutant plant large subunit of AGPase. In one embodiment, a polynucleotide of the invention encodes a plant AGPase large subunit having an amino acid mutation wherein the cysteine amino acid corresponding to amino acid position 424 of wild type maize AGPase large subunit is substituted with an amino acid that results in increased AGPase activity when provided in an AGPase enzyme. In a specific embodiment, the cysteine corresponding to amino acid position 424 of wild type maize AGPase large subunit is substituted with a valine amino acid. The subject invention also comprises a mutant plant large subunit of AGPase encoded by a polynucleotide of the invention. Characterization of kinetic and allosteric properties indicates increased starch yield is provided when the polynucleotides of the invention are expressed in plants such as monocot endosperms.

Methods and materials for increasing starch biosynthesis in plants

Nucleic acids encoding heat stable mutants of plant ADP-glucose pyrophosphorylase

The subject invention concerns chimeric AGP subunit proteins and polynucleotides that encode the chimeric proteins. The subject invention provides for mutant AGP enzymes comprising a chimeric subunit of the invention that are less sensitive to inorganic phosphate than wild type AGP enzymes. In one embodiment, the AGP subunit is a small subunit of a plant AGP enzyme. The subject invention also concerns plants comprising a polynucleotide encoding a chimeric AGP subunit protein of the invention. The subject invention also concerns methods for producing a plant comprising a polynucleotide of the present invention. Plants produced according to the invention comprise AGP enzymes that are less sensitive to inorganic phosphate than wild type AGP enzyme.

Variants of ADP-glucose pyrophosphorylase affecting phosphate sensitivity and other parameters

The subject invention pertains to novel mutant polynucleolide molecules that encode enzymes that have increased heat stability. These polynucleotides, when expressed in plants, result in increased yield in plants grown under conditions of heat stress. In one embodiment, the polynucleotide molecules of the subject invention encode maize endosperm ADP glucose pyrophosphorylase (AGP) activity. Plants and plant tissue bred to contain, or transformed with, the mutant polynucleotides, and expressing the polypeptides encoded by the polynucleotides, are also contemplated by the present invention. The subject invention also concerns methods for isolating polynucleotides and polypeptides contemplated within the scope of the invention. Methods for increasing yield in plants grown under conditions or heat stress are also provided.

The subject invention concerns materials and methods for enhancing starch production in plants. Starch production is enhanced, relative to levels observed in wildtype or control plants, by reduction of the plant 14-3-3 protein(s) which subsequently results in increased accumulation of starch in the plant. In one embodiment, the 14-3-3 protein expression is reduced using polynucleotides that are antisense to the 14-3-3 gene sequences expressed in the plant. In another embodiment, the 14-3-3 protein expression is reduced by 'knockout' of a 14-3-3 gene or gene sequences. The subject invention also pertains to transformed and transgenic plants that have polynucleotides that are antisense to the 14-3-3 gene sequences expressed in the plant, wherein the transformed and transgenic plants exhibit enhanced starch production. The subject invention also pertains to 'knockout' plants in which the normal functional 14-3-3 gene in the plant is deleted or replaced with a non-functional form of the gene. The subject invention also concerns the &#x201c;antisense&#x201d; polynucleotides of the invention that when introduced into a plant cell can function to effectively reduce expression of the 14-3-3 proteins in a plant.

Plants with enhanced ability to produce starch and methods for obtaining them

The subject invention pertains to novel mutant polynucleotide molecules that encode enzymes that have increased heat stability. These polynucleotides, when expressed in plants, result in increased yield in plants grown under conditions of heat stress. The polynucleotide molecules of the subject invention encode maize endosperm ADP glucose pyrophosphorylase (AGP) and soluble starch synthase (SSS) enzyme activties. Plants and plant tissue bred to contain, or transformed with, the mutant polynucleotides, and expressing the polypeptides encoded by the polynucleotides, are also contemplated by the present invention. The subject invention also concerns methods for isolating polynucleotides and polypeptides contemplated within the scope of the invention. Methods for increasing yield in plants grown under conditions of heat stress are also provided.

The present invention pertains to mutant alleles of the genes encoding the large subunit of AGP-glucose pyrophosphorylase in plants. When present in maize plants, these mutant alleles confer enhanced germination characteristics but without any dimishment of food quality or flavor as compared to plants that express the sh2-R genes. The subject invention also concerns methods for transforming plants with the mutant alleles. The subject invention also concerns plants that have the mutant alleles of the invention incorporated in their genome.

Mutant genes encoding plant ADP-glucose pyrophosphorylase and methods of use

The subject invention pertains to novel mutant polynucleotide molecules that encode enzymes that have increased heat stability. These polynucleotides, when expressed in plants, result in increased yield in plants grown under conditions of heat stress. The polynucleotide molecules of the subject invention encode maize endosperm ADP glucose pyrophosphorylase (AGP) and soluble starch synthase (SSS) enzyme activities. Plants and plant tissue bred to contain, or transformed with, the mutant polynucleotides, and expressing the polypeptides encoded by the polynucleotides, are also contemplated by the present invention. The subject invention also concerns methods for isolating polynucleotides and polypeptides contemplated within the scope of the invention. Methods for increasing yield in plants grown under conditions of best stress are also provided.

The present invention provides a novel means for enhancing the expression of genes by the use of the first intron of the Zea mays Sh1 cDNA, functional sequences derived therefrom and/or sequences flanking the first Sh1 intron, placed between the transcription start site and the translation start site of the coding sequence for enhanced expression is sought. The DNA sequences provided herein can be used, for example, to increase protein expression in engineered plant cells or in transgenic plants.

Means for enhancing gene expression

The subject invention pertains to novel variants of the maize gene, Shrunken2 (Sh2) and a method of using that gene. The variant gene, Sh2-m1Rev6, encodes a subunit of the ADP-glucose pyrophosphorylase (AGP) enzyme that has additional amino acids inserted in or near the allosteric binding site of the protein. Corn seed expressing the Sh2-m1Rev6 gene has a 15% weight increase over wild type seed. The increase in seed weight is not associated simply with an increase in percentage starch content of the seed.

Growing community of inventors

Gainesville, FL, United States of America

L Curtis Hannah