Wheat Haploid Inducer Plant NLD Gene Inhibition
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Solution Overview
Problem
Current methods for producing homozygous lines in wheat breeding are labor-intensive and time-consuming, and there is a lack of efficient haploid inducer lines in wheat, unlike in maize, which hampers rapid genetic improvement and gene stacking.
Innovation Solution
Development of a wheat haploid inducer plant with non-functional alleles of the NLD genes and a dominant or semi-dominant genetic marker to quickly identify and sort haploid progeny, utilizing site-directed mutagenesis, CRISPR-Cas systems, and RNAi constructs to inhibit NLD gene expression and introduce genetic markers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional recurrent selfing is used to establish homozygous lines, then homozygosity is achieved, but the process requires 8-10 generations and is time-consuming
Solution Approach 1:
The invention changes the ploidy parameter of the plant from diploid to haploid through induced haploidization. By treating plant cells or tissues with colchicine or other haploid-inducing agents, the chromosome number is reduced by half, directly producing haploid plants that are automatically homozygous for all genes, thus achieving complete homozygosity in one step rather than requiring multiple generations of selfing.
2Productivity
If in vitro culture of anthers or microspores is used to produce doubled haploids, then homozygous lines are obtained faster, but the process is labor-intensive and genotype-dependent
Solution Approach 1:
The invention extracts and utilizes specific endosperm cells or embryo cells from the seed that exhibit haploid characteristics. By isolating these cells and culturing them separately, the method bypasses the need for complex whole-plant in vitro culture systems. This extraction approach simplifies the procedure, reduces labor intensity, and makes the process applicable to various genotypes without requiring optimization for each specific variety.
3Productivity
If corn pollen is used to pollinate wheat spikes to produce doubled haploids, then haploid induction occurs, but embryo rescue is required and the process is time-consuming
Solution Approach 1:
The invention employs wheat pollen that carries endosperm-specific markers (such as storage protein genes like Gli-1 or Glu-1) to self-mark the resulting embryos. These markers allow direct visual or molecular identification of haploid embryos within the seed without requiring embryo rescue or complex external detection systems. The system serves itself by using the plant's own genetic markers to indicate haploid status, eliminating the need for additional technical interventions.
4Measurement precision
If molecular markers are used to identify haploid progeny, then identification accuracy is improved, but detection complexity and cost increase
Solution Approach 1:
The invention utilizes endosperm-specific genetic markers that result in visible color or pigment differences between haploid and diploid seeds. For example, haploid seeds may lack certain pigments or display different coloration patterns compared to diploid seeds, allowing for simple visual screening. This color-based detection method maintains high identification accuracy while avoiding the complexity and cost of molecular marker analysis, as the phenotypic expression occurs at the seed level and can be observed without specialized equipment.
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 enables rapid identification and production of haploid plants, accelerating the breeding process and improving genetic variance in wheat, reducing the time and labor required for establishing homozygous lines.
Implementation Method 1
utilizing site-directed mutagenesis, CRISPR-Cas systems, and RNAi constructs to inhibit NLD gene expression
Implementation Method 2
utilizing site-directed mutagenesis, CRISPR-Cas systems, and RNAi constructs to inhibit NLD gene expression
Data Source
AI summary
The invention relates to a wheat haploid inducer plant comprising at least one cell which presents inhibition of the expression of the three NLD genes of genome A, B and D, and at least one dominant or semi-dominant genetic marker, wherein said genetic marker produces, a detectable phenotype, as well as methods of uses.