5 results about "Als gene" patented technology

This invention belongs to the fields of applied environmental microbiology and agricultural biotechnology, and discloses ImALS, an acetyllactone synthase resistant to high concentrations of methoxylamine, its encoding gene, and its applications. Its nucleotide sequence is SEQ ID NO.1, with a full length of 1785 bp, encoding 594 amino acids, and its amino acid sequence is SEQ ID NO.2. ImALS shares less than 58.3% homology with previously reported acetyllactone synthases, thus representing a novel ALS gene. The acetyllactone synthase ImALS provided by this invention exhibits a specific activity of 0.52 mM / mg against sodium pyruvate. ImALS demonstrates strong resistance to the ALS inhibitor methoxylamine herbicide, with a half-maximal inhibitory concentration (IC50) of 1500 μM, indicating that the ImALS gene has significant application potential in methoxylamine-resistant transgenic engineering.

The application discloses a rapid molecular detection method for a His-170-Pro mutation of a green dogtail grass ALS gene, and belongs to the technical field of gene mutation detection. First, a DNA sample of the green dogtail grass is extracted, then LAMP specific primers designed according to a resistant mutation site of the ALS gene are used for constant-temperature amplification, a fluorescent dye is added into the amplification product, the fluorescent intensity change is observed under ultraviolet light, and / or the amplification product is subjected to agarose gel electrophoresis analysis, so that the rapid molecular detection of the His-170-Pro mutation of the green dogtail grass ALS gene is realized. The application realizes rapid, efficient and visual detection of the target nucleic acid, overcomes the defects of time-consuming, laborious, high cost, high dependence on instruments and equipment, complex procedures, long detection period and accuracy easily affected by operation in the prior art ALS-resistant green dogtail grass His-170-Pro mutation identification and detection method, and can greatly improve the field detection efficiency of the resistant green dogtail grass.

The present application relates to the technical field of nucleotide molecule screening, in particular to a mutagenesis screening method and application of a rice herbicide-resistant genotype. The present application designs a qPCR reaction system based on a specific SNP site region of a mutant ALS gene, realizes the effect of high-throughput and large-scale screening of a rice parent containing a mutant ALS gene, and can significantly improve the efficiency of rice screening and breeding. The present application designs a TaqMan-MGB probe based on a specific SNP site region of a mutant ALS gene, which can form a hybrid molecule with a specific SNP site with high specificity and high accuracy, ensuring the accuracy and reproducibility of qPCR.

This invention belongs to the field of molecular biology detection technology, and provides a primer composition, gDNA, probe, and method for detecting amino acid mutation sites in ALS of Amaranthus retroflexus. This invention is the first to construct a method for detecting multiple resistance mutations at multiple sites based on the synergistic effect of multiplex PCR and PfAgo. Through multiplex PCR amplification and PfAgo specific recognition and cleavage, it achieves simultaneous detection and mutation type identification of multiple resistance target sites in the ALS gene of Amaranthus retroflexus. Based on the multiple constraint mechanism of multiplex PCR amplification, PfAgo guided sequence recognition, and fluorescent probe cleavage, it achieves stepwise identification and verification of target sites, exhibiting excellent specificity, sensitivity, and robustness, reducing the risk of false positives, ensuring accurate and reliable results, and requiring no standards for interpretation. It is suitable for rapid and simultaneous multi-site detection of resistance to ALS inhibitors in Amaranthus retroflexus in the field, is simple to operate, and has low cost, providing technical support for resistance monitoring and precision herbicide management.

The present application belongs to the technical field of crop breeding, and relates to a potato breeding method for editing ALS gene. The method comprises the following steps: (1) using agrobacterium transformed with a gene editing tool to infect potato explants, wherein the infection medium used contains CaCl2 with a total concentration of 9-15 mM, and the gene editing tool targets potato ALS gene for editing mutation of ALS gene to make potato resistant to imidazolinone herbicides; (2) after co-cultivation and recovery culture, the potato explants are sequentially subjected to resistant callus induction, resistant bud induction and seedling culture on resistant callus induction medium, resistant bud induction medium and seedling medium to obtain regenerated seedlings, and the resistant callus induction medium, resistant bud induction medium and seedling medium all contain imidazolinone herbicides. By using the method of the present application, potato varieties with ALS gene mutation can be obtained with high infection efficiency and high gene editing efficiency.