Method for high throughput transgene function analysis for agronomic traits in maize
AssigneePioneer Hi-Bred International, Inc.
InventorsJONATHAN LIGHTNER, IGOR OLIVEIRA, JON MUSGRAVE
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Abstract
A method for the rapid evaluation of transgene function in maize plants. The method combines high throughput gene construction methods and high efficiency plant transformation techniques in a specifically developed germplasm. In one as aspect, the method uses quantitative, non-destructive imaging technology applied in a portion or throughout the entire life cycle of a test plant to evaluate agronomic traits of interest in a controlled, statistically relevant greenhouse environment. The method reports transgene function early in the transgenic variety development process, eliminating the need to generate seed necessary for multi-location replicated field trials.
Claims28
A method for the rapid analysis of the effect of a transgene of interest on a selected phenotypic parameter in corn, the method comprising the steps of:
- providing a population of transgenic corn plants grown in an environmentally controlled greenhouse environment;
- providing a light spectrum digital analyzing device;
- using the light spectrum digital analyzing device to take original reflected light images of each plant and then analyze those images to determine a value for the phenotypic parameter in each plant;
- and
- calculating a mean or median value of the phenotypic parameter and the biological variance for the population of transgenic plants;
- whereby any statistically significant difference between the phenotypic parameter value for a single member of the population of transgenic plants and the mean or median value for the population of transgenic plants may be considered attributable to the transgene of interest.
A method for the high throughput analysis of a yield component trait in transgenic corn, the method comprising the steps of:
- providing recipient maize plant cells from a plant line for transformation;
- obtaining a plurality of nucleotide vectors;
- introducing the plurality of nucleotide vectors into the recipient maize plant cells to create a plurality of transgenic plants;
- allowing the plurality of transgenic plants and, optionally, a plurality of control plants, to grow in an environmentally controlled greenhouse environment, each plant being associated with a machine-readable label that distinguishes the plant from other plants;
- analyzing each transgenic plant and, optionally, each control plant, for the yield component trait using quantitative, non-destructive imaging technology and associating the yield component trait with the machine-readable information number for that plant;
- and
- comparing the yield component trait for one or more transgenic plants with the mean or median yield component trait of transgenic plants, or optionally, with the mean or median yield component trait for the control plants, to determine the effect of the nucleotide vectors on the yield component trait.
A method for the rapid analysis of the effect of a gene of interest on a selected phenotypic parameter in corn, the method comprising the steps of:
- providing a population of T0 transgenic corn plants possessing the gene of interest and grown in an environmentally controlled greenhouse environment;
- using a light spectrum digital analyzing device having an instrumental variance below about 5% to take original reflected light images of each plant and then analyze those images to obtain phenotypic data for each plant;
- and
- performing a statistical analysis of the phenotypic data to analyze the effect of the gene of interest on the selected phenotypic parameter.
A method for the rapid analysis of the effect of a gene of interest on a selected phenotypic parameter in corn, the method comprising the steps of:
- providing a population of T0 transgenic corn plants possessing the gene of interest and grown in an environmentally controlled greenhouse environment;
- pollinating the T0 transgenic corn plants with a recurrent parent to obtain T1 plants;
- assaying the T1 plants to identify plants possessing the gene of interest;
- using a light spectrum digital analyzing device having an instrumental variance below about 5% to take original reflected light images of each T1 plant and then analyze those images to obtain phenotypic data for each plant;
- and
- performing a statistical analysis of the phenotypic data to analyze the effect of the gene of interest on the selected phenotypic parameter.
Dependencies are inferred from the claim text.