CENH3 Haploid Inducer Mutations Without Transgenic Modification
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Solution Overview
Problem
Existing methods for generating haploid plants require substantial modifications of the CENH3 protein or involve genetic engineering, which are economically and socially problematic due to public reluctance and regulatory challenges.
Innovation Solution
Non-transgenic methods are employed to alter the amino acid sequence of the CENH3 protein in specific domains, such as the N-terminal tail, aN-helix, α1-helix, loop1, α2-helix, loop2, α3-helix, and C-terminal domain, to induce haploid production in plants, using mutations that confer the biological activity of a haploid inducer.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If substantial modifications of the CENH3 protein are used to generate haploid plants, then haploid production efficiency is improved, but genetic engineering requirements increase causing economic and social problems
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the CENH3 protein sequence to create haploid-inducing variants. By changing parameters such as amino acid substitutions at specific positions (e.g., R8K, R10F, R11K), the patent achieves haploid production without requiring complex genetic engineering approaches like GFP fusion or tailswap modifications, thus resolving the contradiction between efficiency and complexity
Solution Approach 2:
The patent applies local quality by making targeted modifications at specific local regions of the CENH3 protein rather than substantial global modifications. By focusing changes on particular amino acid positions within the protein sequence, the patent achieves the desired haploid-inducing function with minimal alterations, reducing the need for complex genetic engineering while maintaining high efficiency
2Reliability
If transgenic CENH3 modifications are employed, then haploid induction capability is achieved, but public acceptance and regulatory approval decrease
Solution Approach 1:
The patent employs simple, minimal modifications to the CENH3 protein sequence that can be achieved through straightforward mutagenesis approaches rather than complex transgenic systems. By using simple amino acid substitutions that can be introduced through relatively simple breeding or mutagenesis programs, the patent reduces the perception of genetic engineering complexity, thereby improving public acceptance and regulatory approval prospects while maintaining reliable haploid induction capability
3Productivity
If multiple amino acid alterations are combined in CENH3, then haploid production efficiency is enhanced, but protein sequence complexity increases
Solution Approach 1:
The patent applies segmentation by dividing the CENH3 protein sequence into distinct regions and making targeted modifications at specific segments or positions. By focusing alterations on particular amino acid residues rather than distributing changes throughout the entire sequence, the patent enhances haploid production efficiency while maintaining the overall stability and structural integrity of the protein composition
Data Source
Figure 1
AI summary
The present invention relates to non-transgenic and transgenic plants, preferably crop plants, having biological activity of a haploid inducer and comprising a polynucleotide which comprises a nucleotide sequence encoding a centromer histone H3 (CENH3) protein, wherein the polynucleotide comprises at least one mutation causing an alteration of the amino acid sequence of the CENH3 protein, and to a part of the part. Further, the invention provides methods of generating the inducer plants, methods of generating haploid and double haploid plants using the inducer plants as well as methods of facilitating cytoplasm exchange.