Apple S gene type rapid identification method and special kit thereof

Abstract

The invention discloses a method for identifying S genotype of an apple and a special reagent kit thereof. The method uses the genomic DNA of an apple variety to be detected as a template and amplification is carried out by the use of primer pairs for identifying the S genotype of the apple variety so as to determine the S genotype of the apple variety to be detected according to the amplification result. By the method, the selfed incompatible S genotype of the apple variety can be fast identified, and pollenizers can be selected through the S genotype thereof. Therefore, the method has important application value on apple planting and hybridization research.

Description

technical field [0001] The invention relates to a rapid identification method of apple S genotype, a special kit and its application in screening apple pollination varieties. Background technique [0002] Apple belongs to the gametophytic self-incompatibility type and is controlled by a single locus of multiple alleles. This locus is also called the S locus, which is located on the 17th pair of chromosomes in the apple genome, on an S locus. Apple populations contain multiple S alleles. The S locus includes the style S gene (S-RNase gene) and the pollen S gene (F-box gene). During self- or cross-pollination, the S allele of the pollen is the same as one of the two S alleles in the style. At the same time, the growth of pollen tubes in the style is inhibited, so that fertilization cannot be completed and self-incompatibility is exhibited (Kao T H, Tsukamoto T. The molecular and genetic bases of S-RNase-based self-incompatibility. Plant Cell, 2004 , 16: S72-S83). The S geno...

AI Technical Summary

Problems solved by technology

The advantage of this method is that it is intuitive and easy to operate, but it also obviously has shortcomings such as large workload, long identification cycle (must be after the apple variety blooms), low efficiency, etc., especially when the S genotype of both parents or one parent is unknown or The S genotype of a breed cannot be determined when the appropriate test breed is not available

Subsequent studies have found that the product encoded by the S gene in the style of apple is a glycoprotein, which has RNase activity. The S genotype can be determined by electrophoresis analysis of the style S glycoprotein. However, due to the complicated operation process, especially isoelectric focusing The technical requirements of polyacrylamide gel electrophoresis are relatively high, and the test conditions are not consistent, and the PI value of each S-RNase may be biased. In addition, the test material used is the style of an old tree, which limits the selection of materials, and A large amount of test material is required, especially when studying hybrid populations, it is necessary to wait for hybrid individuals to pass through childhood

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