Transgenic plants with modified sugar content and methods of generating same
This application provides and discloses small RNAs and their target genes that are involved in controlling the levels of sucrose, glucose, and fructose and methods of modulating expression or activity of these mi RNAs and target genes. This application further provides transgenic plants, plant parts, e.g., seeds, that have altered expression of these mi RNAs and target genes and have increased levels of sucrose, increased sucrose to glucose ratios, increased sucrose to hexose ratios, altered carbohydrate levels, or increased Brix in fruit from transgenic plants. This application also provides methods of producing and growing transgenic plants or seeds that have increased levels of sucrose, increased sucrose to glucose ratios, increased sucrose to hexose ratios, altered carbohydrate levels, or increased Brix in fruit from transgenic plants.
1. A method of increasing the sucrose level or increasing the sucrose to glucose ratio in a tomato plant comprising expressing in said tomato plant
a DNA encoding a miR397-, miR528-, or miR1110-resistant target gene, wherein said miR397-, miR528-, or miR1110-resistant target gene comprises an introduced silent mutation in a nucleotide sequence that is otherwise substantially identical to the nucleotide sequence of an endogenous gene that is natively regulated by miR397, miR528, or miR1110, and wherein said silent mutation prevents binding by a mature miR397, miR528, or miR1110 to a transcript of said miR397-, miR528-, or miR1110-resistant target gene,
wherein the sucrose level or the sucrose-to-glucose ratio is increased in said tomato plant.
2. The method of claim 1 , wherein said at least one DNA
comprises a miR397-resistant target gene encoding an amino acid sequence having at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 503-512.
3. The method of claim 1 , wherein said method achieves a tomato fruit comprising more than 0.5%, more than 0.75%, more than 1%, more than 1.5%, more than 2%, more than 2.5%, more than 3%, more than 3.5%, or more than 4% sucrose by fresh weight.
4. The method of claim 1 , wherein the heterologous promoter is a constitutive promoter or an inducible promoter.
5. The method of claim 1 , wherein the heterologous promoter is a CaMV 35S promoter or a tomato fruit specific promoter.
6. A method of modifying the carbohydrate content in an edible plant comprising expressing in said edible plant at least one DNA selected from the group consisting of
a DNA encoding a miR397-, miR528-, or miR1110-resistant target gene, wherein said miR169-, miR397-, miR528-, or miR1110-resistant target gene comprises an introduced silent mutation in a nucleotide sequence that is otherwise substantially identical to the nucleotide sequence of an endogenous gene that is natively regulated by miR169, miR397, miR528, or miR1110, and wherein said silent mutation prevents binding by a mature miR169, miR397, miR528, or miR1110 to a transcript of said miR169-, miR397-, miR528-, or miR1110-resistant target gene,
wherein the carbohydrate content of said edible plant is modified.
7. The method of claim 6 , wherein said at least one DNA
comprises a miR397-resistant target gene encoding an amino acid sequence having at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 503-512.
8. The method of claim 6 , wherein said edible plant comprises more than 0.5%, more than 0.75%, more than 1%, more than 1.5%, more than 2%, more than 2.5%, more than 3%, more than 3.5%, or more than 4% sucrose in a fruit by fresh weight.