Sugarcane bacilliform viral enhancer-based activation tagging platform for maize, and resultant tagged populations and plants
View Patent ↗An activation tagging construct for maize that includes one or more sugarcane bacilliform viral (SCBV) enhancer elements, and resulting tagged populations and plants, are described. In one example, an activation tagging DNA construct includes a coding sequence for a transposase, a detectable reporter (such as anthocyanin regulatory genes B-Peru and C1) and a non-autonomous transposable T-DNA cassette. For example, the transposable T-DNA cassette is inserted into the detectable reporter encoding region such that the B-Peru and C1 genes express anthocyanins in a cell containing the maize activation tagging DNA construct only upon excision of the transposable cassette. Methods of generating a tagged population of maize plants include transforming a maize plant cell or tissue with the disclosed constructs.
1. A method, comprising:
randomly integrating an activation tagging construct into one or more maize plant chromosomes to thereby generate one or more tagged maize plants;
screening the one or more tagged maize plants to identify one or more maize plants having a modified phenotype relative to an untagged maize plant lacking the activation tagging construct; and
selecting the one or more tagged maize plants identified as having a modified phenotype, wherein the modified phenotype is an enhanced agronomic trait,
wherein the activation tagging construct comprises at least one sugarcane bacilliform viral (SCBV) enhancer element set forth in SEQ ID NO: 2, and the enhancer element is operatively linked to a promoter that is native to the maize plant.
2. The method of claim 1 , comprising generating a tagged population of maize plants.
3. The method of claim 1 , further comprising identifying a tagged population of maize plants.
4. The method of claim 1 , wherein the activation tagging construct comprises a selectable marker and the method further comprises selecting a tagged population of maize plants based on the presence of the selectable marker in the tagged population of maize plants.
5. The method of claim 1 , wherein the activation tagging construct is randomly integrated by transformation.
6. The method of claim 5 , wherein the activation tagging construct is randomly integrated by Agrobacterium mediated transformation.
7. The method of claim 1 , wherein the activation tagging construct is randomly integrated by transposition.
8. The method of claim 1 , wherein the activation tagging construct comprises at least two copies of the SCBV enhancer.
9. The method of claim 8 , wherein the activation tagging construct comprises four copies of the SCBV enhancer.
10. The method of claim 1 , wherein the activation tagging construct comprises the sequence of SEQ ID NO: 1.
11. The method of claim 1 , wherein the enhanced agronomic trait comprises increased drought tolerance, increased nitrogen use efficiency, increased seed oil content, or increased yield under an environmental stress condition.
12. The method of claim 11 , wherein the environmental stress condition comprises drought, shade, fungal disease, viral disease, bacterial disease, insect infestation, nematode infestation, cold temperature exposure, heat exposure, osmotic stress, reduced nitrogen nutrient availability, reduced phosphorus nutrient availability and high plant density.