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Apple S gene type rapid identification method and special kit thereof

A genotype and kit technology, applied in the field of screening apple pollinated varieties, can solve problems such as complex operation process, inconsistent test conditions, long identification cycle, etc., to simplify experimental operation, save identification time, and achieve major applications value effect

Inactive Publication Date: 2009-03-25
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

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  • Apple S gene type rapid identification method and special kit thereof
  • Apple S gene type rapid identification method and special kit thereof
  • Apple S gene type rapid identification method and special kit thereof

Examples

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Embodiment 1

[0036] Embodiment 1, identification of apple S genotype

[0037] 1. Amplification of Apple S gene-specific fragments

[0038] After sequence analysis of the primary structure of each apple S-RNase gene, specific primers for each S gene were designed in the specific regions of each gene sequence. The primer sequences and the specific fragment sizes predicted by each primer are shown in Table 1.

[0039] Table 1. Apple S gene-specific primers

[0040]

[0041] Using the above primers respectively, each S gene-specific fragment was amplified from apple varieties of known S genotype, and from apple variety Fuji (Fuji) (S 1 S 9 ) in amplified S 1 , S 9 Gene-specific fragment; from apple variety Golden Delicious (S 2 S 3 ) in amplified S 2 and S 3 Gene-specific fragments from the apple cultivar Gravenstein (S 4 S 11 S 20 ) in amplified S 4 Gene-specific fragments from apple variety Gala (Gala) (S 2 S 5 ) in amplified S 5 Gene-specific fragments from apple variety H...

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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 and its special kit and its application in screening apple pollinated varieties. Background technique [0002] Apple belongs to the type of gametophytic self-incompatibility, which is controlled by the multiple alleles of a single locus. This locus is also called the S locus. It is located on the 17th pair of chromosomes in the apple genome, at an S locus Apple populations contained multiple S alleles. The S locus includes the style S gene (S-RNase gene) and the pollen S gene (F-box gene). During self-flowering 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 will be inhibited, so that fertilization cannot be completed and self-incompatibility will be displayed (Kao T H, Tsukamoto T. The molecular and genetic bases of S-RNase-based self-incompatibility. Plant Cell, 20...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 李天忠龙慎山韩振海
Owner CHINA AGRI UNIV
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