A method for testing the specificity, consistency and stability of new pure-line rapeseed varieties

Abstract

The invention discloses a method for testing the specificity, consistency and stability of a new pure-line rapeseed variety. The method comprises: obtaining variation sites; determining the test area of ​​the rapeseed variety to be tested; constructing a database; after determining the sampling amount, randomly sampling and mixing and extracting the DNA of the mixed sample; preparing primers; using the primers to amplify the DNA of the mixed sample , the amplified product is used to construct a high-throughput sequencing library; perform high-throughput sequencing on the high-throughput sequencing library to obtain a sequenced fragment group; analyze the sequenced fragment group to obtain the genotype of the rapeseed variety to be tested and the genotype of the hybrid plant; compare the obtained Approximate variety, variation site and rate of variation site; compare the genotype of the hybrid with the genotype in the database, and calculate the rate of hybrid after obtaining the variety; Judging the specificity, consistency and stability of the rape variety to be tested. The method can accurately and completely judge the specificity, stability and consistency of the rape varieties to be tested.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for testing the specificity, consistency and stability of a new pure-line rapeseed variety. Background technique [0002] As a specialized intellectual property, new plant varieties have become the core competitiveness of a company or even a country. The authorization of new plant varieties and the resolution of related legal issues rely on DUS testing, which is the field planting identification or indoor molecular marker identification of the specificity (Distinctness), uniformity (Uniformity) and stability (Stability) of the rape variety to be tested. The field planting identification process is as follows: plant the rapeseed variety to be tested and similar varieties in the field at the same time, observe their multiple traits in the growing season of 2 years or more, and judge the significant difference between the rapeseed variety to be tested and similar varieties base...

AI Technical Summary

Problems solved by technology

[0003] The disadvantages of field planting identification are: long cycle, heavy workload, and environment affecting traits, resulting in inaccurate judgment

The disadvantages of indoor molecular marker identification are: each test area of ​​each sample needs to be processed separately, the workload is heavy, a large number of samples and test areas cannot be sampled, and the rate of hybrid plants cannot be calculated, so stability and consistency tests cannot be performed

The common disadvantage of field planting identification and indoor molecular marker identification is that due to the workload, it is impossible to objectively select similar varieties from existing varieties, which can only be provided by the variety right applicant. Approximate breeds provided by others may not be true, resulting in legal consequences of wrong breed authorization

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