Rapid identification primers of citrus hybrids based on SNP markers as well as method

Abstract

The invention provides rapid identification primers of citrus hybrids based on SNP markers as well as a method. The method comprises the following steps: 1), the parent DNA of a hybrid citrus is extracted and used as a template, P29488785F / P28489624R are used as primers for PCR amplification, the length of amplified fragment is 858 bp, Sanger sequencing is performed on the amplified products, andSNP marker sites between parents are analyzed; and 2), genomic DNA from leaf tissue of hybrid offspring is extracted and used as a template, P29488785F / P29489624R are used as the primers, PCR is performed for obtaining the amplified products, and P29488785F are used as sequencing primers to perform Sanger sequencing on the amplified products. In the analysis of a sequencing peak map, if impurity peaks appear at the SNP sites, the products are true hybrids, and the hybrid identification work is completed. The steps in the method are simple and can be applied to rapid identification of hybrid offspring on a large scale. Furthermore, the consequence of identification is accurate and reliable.

Description

technical field [0001] The invention relates to a method for identifying hybrid varieties among plants, in particular to a primer and method for rapid identification of citrus hybrids based on SNP markers. Background technique [0002] Citrus is highly heterozygous genetically, and most of the main citrus varieties are polyembryonic, which makes it difficult to obtain hybrids through sexual hybridization. Hybrid rescue can preserve the immature embryos of true hybrids, but a large number of nucellar embryos also survive and develop Breeding seedlings, how to identify true hybrids from hybrid offspring is a major problem that plagues citrus breeding. At present, citrus hybrid identification mainly uses methods such as morphology, cytology, isozyme, random amplified polymorphic DNA (RAPD), SSR markers and molecular hybridization. Morphological methods are greatly affected by environmental conditions, and the accuracy is poor; chromosome counting cannot distinguish homologous ...

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Problems solved by technology

Morphological methods are greatly affected by environmental conditions, and the accuracy is poor; chromosome counting cannot distinguish homologous and heterologous fusion tetraploids; isozymes can only indirectly infer gene changes, and there are fewer detection sites , physiological state, environmental conditions, etc.; RAPD technology is affected by many factors such as the combination of random primers and templates, the specificity is not strong, and the repeatability of the results is not very good; The limitation of the new alleles generated, the accidental slippage during DNA replication will also lead to changes in the number of repeat units; molecular hybridization is an effective method for the identification of somatic cell hybrids, but it requires high DNA quality, a large amount of use, and the analysis process is cumbersome

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