Screening and detection method based on nuclear male sterility genes of oil flax

By using single-cell transcriptome sequencing and high-resolution melting curve detection, the problem of gene expression differential identification and detection in the screening of male sterility genes in oil flax cells has been solved, achieving efficient and accurate genotype differentiation, and is suitable for rapid screening of large batches of breeding materials.

CN122445768APending Publication Date: 2026-07-24INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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Patent Information

Application Number
CN202610825610.8
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-06-09
Publication Date
2026-07-24

AI Technical Summary

Technical Problem

Existing technologies struggle to capture the subtle differences in gene expression before and after meiosis in screening for male sterility genes in oil flax cells, and to identify candidate genes associated with functional abnormalities in tapetum cells and microsporocytes. Furthermore, the detection methods often fail to balance high throughput with accuracy.

Method used

Single-cell transcriptome sequencing combined with high-resolution melting curve detection was employed. Differentially expressed genes in cell populations were identified through dimensionality reduction clustering analysis. High-resolution melting curve detection primers were designed to distinguish genotypes, avoiding the need for opening amplification products and achieving rapid and accurate genotype differentiation.

Benefits of technology

It improves the accuracy and efficiency of screening for male sterility genes in oil flax cells, reduces false positive results, simplifies the operation process, avoids cross-contamination, and is suitable for genotype screening of large batches of breeding materials.

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Abstract

The invention discloses a screening and detecting method based on nuclear male sterility genes of oil flax, and belongs to the technical field of molecular biology and plant breeding. The method comprises the following steps: carrying out single cell transcriptome sequencing on anther tissues of the oil flax flower buds in multiple development periods, and identifying cell population differential expression genes of tapetum cells and microspore mother cells before meiosis; carrying out homologous comparison on the cell population differential expression gene and an arabidopsis thaliana cell nucleus male sterility gene set, and screening oil flax cell nucleus male sterility candidate genes; designing an overlapped primer pair aiming at the candidate genes, and carrying out long-fragment PCR amplification and sequencing to obtain sequence difference sites between the sterile line and the fertile line; designing a high-resolution melting curve detection primer group according to the sequence difference sites to perform amplification reaction, and distinguishing the genotypes of the sterile line and the fertile line according to the peak pattern difference of the melting curve.
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