Methods for controlling gibberellin levels
Methods and materials are disclosed for the inhibition and control of gibberellic acid levels. In particular, nucleic acid sequences of copalyl diphosphate synthase. 3-β hydroxylase, and 2-oxidase and additional nucleic acid sequences are disclosed. Gibberellic acid levels may be inhibited or controlled by preparation of a chimeric expression construct capable of expressing a RNA or protein product which suppresses the gibberellin biosynthetic pathway sequence, diverts substrates from the pathway or degrades pathway substrates or products. The sequence is preferably a copalyl diphosphate synthase sequence, a 3β-hydroxylase sequence, a 2-oxidase sequence, a phytoene synthase sequence, a C20-oxidase sequence, and a 2β,3β-hydroxylase sequence. Administration of a complementing agent, preferably a gibberellin or gibberellin precursor or intermediate restores bioactivity.
1. A method of growing a transgenic plant comprising:
providing a transgenic plant or a seed or seedling thereof comprising a transgene, the transgene comprising a promoter and, operably linked to the promoter, a sequence that encodes an enzyme that inactivates an endogenous gibberellin in a plant cell, causing reduced levels of the endogenous gibberellin in the plant cell, compared with an otherwise identical plant or seed or seedling thereof that lacks the transgene, wherein the sequence is SEQ ID NO: 62;
applying a composition that comprises at least one GA compound that is metabolized by the seed or seedling to produce a product having gibberellin activity that is not inactivated by the enzyme to the transgenic plant or the seed or seedling thereof; and
growing the transgenic plant or the seed or seedling thereof to produce a phenotypically normal transgenic plant.
2. The method of claim 1 , wherein the application of said GA compound is to the soil in which the transgenic plant or seed or seedling thereof is grown.
3. The method of claim 1 , wherein the sequence encodes the soybean GA 2-oxidase 1 (SEQ ID NO:63).
4. The method of claim 1 , wherein the promoter is preferentially expressed in developing seeds, during seed germination, or in early seedlings.
5. The method of claim 4 , wherein said promoter is selected from the group consisting of the Brassica isocitrate lyase gene promoter, the soybean AX5 gene promoter (SEQ ID NO:7), the soybean Sle2 gene promoter, and the At2ox4 gene promoter.
6. The method of claim 1 , wherein the GA compound is GA 4 , GA 7 , GA 3 , or GA 3 -3-acetate.
7. The method of claim 6 , wherein the GA compound is GA 3 or GA 3 -3-acetate.
8. An isolated nucleic acid construct comprising a promoter that causes expression of an operably linked nucleic acid segment in a plant cell and, operably linked to the promoter, the nucleic acid segment comprising a sequence that encodes a polypeptide having a GA 2-oxidase activity, wherein expression of the nucleic acid segment in the plant cell results in inactivation of an endogenous gibberellin in the plant cell, thereby reducing levels of the endogenous gibberellin in the plant cell compared with an otherwise identical plant cell in which the nucleic acid segment is not expressed, wherein the sequence is SEQ ID NO: 62.
9. A transgenic plant transformed with a nucleic acid segment, wherein the nucleic acid segment comprises a promoter that causes expression of an operably linked nucleic acid segment in a plant cell and , operably linked to the promoter, the nucleic acid segment comprising a sequence that encodes a polypeptide having a GA 2-oxidase activity, wherein the sequence is SEQ ID NO: 62, and expression of the nucleic acid segment in the plant cell results in inactivation of an endogenous gibberellin in the plant cell, thereby reducing levels of the endogenous gibberellin in the plant cell compared with an otherwise identical plant cell in which the nucleic acid segment is not expressed.
10. The transgenic plant of claim 9 , characterized by at least one phenotype selected from the group consisting of a shortened hypocotyl, shortened epicotyl, and both a shortened hypocotyl and shortened epicotyl compared with an otherwise identical plant that lacks the nucleic acid segment.