Non-transgenic tomato varieties having increased shelf life post-harvest due to alterations in β-galactosidase 4
The present invention provides a series of independent human-induced, non-transgenic mutations found in at least one tomato β-galactosidase 4 gene, tomato plants having these mutations in at least one of their tomato β-galactosidase 4 genes, and a method of creating and identifying similar and/or additional mutations in the tomato β-galactosidase 4 gene by screening pooled and/or individual tomato plants. Tomato plants identified and produced in accordance with the present invention have fruit that are firmer when ripe with reduced post-harvest softening compared to fruit from wild type tomato plants as a result of non-transgenic mutations in at least one of their tomato β-galactosidase 4 genes.
1. A method of identifying a tomato plant that produces fruit that is firmer when ripe than fruit from wild type tomato plants, said method comprising the steps of:
(a) obtaining plant material from a parent tomato plant;
(b) treating said plant material with a mutagen to create mutagenized plant material;
(c) culturing said mutagenized plant material to produce progeny tomato plants; and
(d) analyzing said mutagenized plant material or said progeny tomato plants to identify a tomato plant having a human-induced, non-transgenic mutation in a tomato β-galactosidase 4 gene having a coding sequence with at least 85% identity to the coding sequence of SEQ ID NO:1, wherein said mutation is selected from the group consisting of C1271T, A2156C and G1981A, wherein the nucleotide change of said mutation is identified through the comparison to SEQ ID NO:1, and wherein said mutation, when expressed in the homozygous state in a tomato plant, causes said tomato plant to produce fruit that is firmer than fruit from wild type tomato plants not having said mutation when ripened at standard storage conditions.
2. The method of claim 1 , wherein said mutation in a tomato β-galactosidase 4 gene codes for a protein comprising the amino acid change P403L and wherein said amino acid change is identified through the comparison to SEQ ID NO: 22.
3. The method of claim 1 , wherein said mutation in a tomato β-galactosidase 4 gene codes for a protein comprising the amino acid change K698T and wherein said amino acid change is identified through the comparison to SEQ ID NO: 22.
4. The method of claim 1 , wherein said mutation in a tomato β-galactosidase 4 gene codes for a protein comprising the amino acid change G640R and wherein said amino acid change is identified through the comparison to SEQ ID NO: 22.
5. A tomato fruit, a seed, a pollen grain, a plant part or a progeny of a tomato plant identified by the method of claim 1 , wherein the fruit, the seed, the pollen grain, the plant part or the progeny comprises the human induced, non-transgenic mutation in the tomato β-galactosidase 4 gene.
6. A food or a food product incorporating the fruit of claim 5 .
7. A method of producing a tomato plant having fruit that is firmer than fruit from a wild type tomato plant, said method comprising the steps of:
(a) screening DNA from at least one tomato plant for mutations in a tomato β-galactosidase 4 gene having a coding sequence with at least 85% identity to the coding sequence of SEQ ID NO:1;
(b) selecting a tomato plant having a mutation in said gene, wherein said mutation is selected from the group consisting of C1271T, A2156C, and G1981A and wherein the nucleotide change of said mutation is identified through comparison to SEQ ID NO:1;
(c) self-pollinating, backcrossing or outcrossing said plant to produce additional progeny plants having said mutation; and
(d) obtaining thereby a tomato plant capable of producing fruit that is firmer than fruit from wild type tomato plants not having said mutation.
8. A tomato fruit, a seed, a pollen gain, a plant part or a progeny of a tomato plant identified by the method of claim 7 , wherein the fruit, the seed, the pollen grain, the plant part, or the progeny comprises the human induced, non-transgenic mutation in the tomato β-galactosidase 4 gene.
9. A food or a food product incorporating the fruit of claim 8 .