AAD-12 Soybean Event Detection for Stable Herbicide Tolerance
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Solution Overview
Problem
The challenge in plant breeding is achieving consistent and stable expression of transgenes in soybeans, as the location of gene insertion in the chromosome influences expression levels and patterns, leading to variability among different events, necessitating the creation and screening of numerous events to identify those with desired transgene expression levels and patterns for herbicide tolerance.
Innovation Solution
The development of the AAD-12 soybean event DAS-68416-4, which includes a specific polynucleotide sequence inserted into a soybean cell genome, providing tolerance to phenoxy auxinic and aryloxyalkanoate herbicides, and methods for detecting this event using unique DNA sequences and PCR assays, allowing for the identification and stacking of herbicide tolerance traits.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If large numbers of events are created and screened to identify events with desired transgene expression levels, then reliable gene expression can be achieved, but the time and resources required for event creation and screening increase significantly
Solution Approach 1:
The patent applies preliminary action by developing and applying event-specific detection methods before full-scale breeding programs. By creating specific PCR assays and molecular markers that can identify events with desired expression characteristics early in the screening process, the invention allows researchers to pre-screen and select promising events without having to create and evaluate large numbers of transformants, thereby saving time and resources while still achieving reliable gene expression
Solution Approach 2:
The patent replaces the mechanical/biological process of creating and phenotypically screening numerous transgenic events with a molecular detection system. Instead of relying on time-consuming growth and expression analysis of multiple events, the invention uses DNA-based detection methods (PCR assays, molecular markers) to rapidly identify events with the desired characteristics, substituting a faster analytical system for the slower biological screening process
2Reliability
If multiple transgenic events are screened to find those with desired expression patterns, then herbicide tolerance reliability improves, but the complexity of the breeding program increases
Solution Approach 1:
The patent extracts the critical identification function from the complex breeding program by developing event-specific detection methods. Instead of managing the complexity of screening multiple events through phenotypic analysis, the invention extracts the essential information (event identity and expression characteristics) through specific molecular assays, simplifying the overall breeding program while maintaining reliability of herbicide tolerance
Solution Approach 2:
The patent introduces molecular markers and event-specific DNA sequences as intermediaries between the transgene integration and the final phenotypic expression. These intermediaries provide a direct link to identify events with desired characteristics without requiring complex breeding operations, thereby reducing program complexity while ensuring reliable herbicide tolerance through selection of appropriate events
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach ensures reliable and stable herbicide tolerance in soybeans by identifying and confirming the presence of the AAD-12 event, enabling the production of soybeans with consistent tolerance to herbicides and facilitating the introgression of this trait into other genetic backgrounds, thereby improving agronomic performance and weed control.
Implementation Method 1
The aad-12 gene (originally from Delftia acidovorans) encodes the aryloxyalkanoate dioxygenase (AAD-12) protein. The trait confers tolerance to 2,4-dichlorophenoxyacetic acid, for example, and to pyridyloxyacetate herbicides.
Implementation Method 2
methods for detecting this event using unique DNA sequences and PCR assays, allowing for the identification and stacking of herbicide tolerance traits
Data Source
AI summary
This invention includes a novel aad-12 transformation event for herbicide tolerance in soybean plants—referred to herein as pDAB4468-0416. This invention includes a heterologous polynucleotide inserted into a specific site within the genome of a soybean cell. In some embodiments, said event/polynucleotide can be “stacked” with other traits, including, for example, other herbicide tolerance gene(s) and/or insect-inhibitory proteins. Additionally, the subject invention provides assays for detecting the presence of the subject event in a sample (a soybean, for example). The assays can be based on the DNA sequence of the recombinant construct, inserted into the soybean genome, and on the genomic sequences flanking the insertion site. Kits and conditions useful in conducting the assays are also provided.