Radish whole-genome SSR core primer combination suitable for SSR-Seq technology and application of radish whole-genome SSR core primer combination

Abstract

The invention belongs to the technical field of biology, particularly relates to the field of plant genetic breeding, and more particularly relates to 38 pairs of radish whole-genome SSR (Simple Sequence Repeat) core primers which are suitable for an SSR-Seq technology and can effectively distinguish various subspecies, variants and varieties of wild radishes and cultivated radishes, a multiplex PCR (Polymerase Chain Reaction) reaction system and a variation identification typing technology of the primers, and integrated application of the primers and the primers. The 38 pairs of primers are uniformly distributed on 9 chromosomes of radishes, are not only suitable for traditional SSR marker detection and have the characteristics of high polymorphism and good repeatability of amplification products, but also can be used for high-throughput and accurate identification of SSR repetitive unit sequences, repetitive unit number and frequency by combining the 38 pairs of SSR primers and a multiplex PCR reaction system thereof provided by the invention with an SSR-seq technology. Efficient molecular detection technical support is provided for research work such as collection and storage, classification and identification, innovation and the like of radish germplasm resources, genetic background analysis of breeding materials and molecular markers of important characters.

Description

technical field [0001] The invention belongs to the field of biotechnology, more specifically the field of plant genetics and breeding, and specifically relates to 38 pairs of radish genome-wide SSR core primers applicable to SSR-Seq technology and capable of effectively distinguishing subspecies, varieties and varieties of wild radish and cultivated radish And the multiple PCR reaction system constructed by it and the application combined with SSR-Seq technology. Background technique [0002] Radish (Raphanus sativus L.2n=18) is a Brassicaceae radish crop, native to the Mediterranean (Georgescu, M.Luchian, V.Groza, O.Ionescu, N.& E. (2016). "Raphanus Raphanistrum Subsp. Landra (Moretti ex DC.) Bonnier & Layens-Adventitious Species of Mediterranean Origin Adapted as Weed in Crops-Some Considerations on Morphological and Anatomical Peculiities". Agriculture and Agricultural Science D. Procedia, 12, 8. Zoh-123 and Hopf, M. (1994). "Domestication of plants in the old world". ...

AI Technical Summary

Problems solved by technology

The previous identification and analysis of radish germplasm resources using SSR markers mostly used ordinary denaturing polyacrylamide gel electrophoresis, and the number of SSR markers developed was limited or not representative of the whole genome. In such cases, it is difficult to assess the efficiency of application of these markers to a wider range of test materials

[0008] Traditional SSR detection methods, such as agarose gel electrophoresis, are cheap, easy to operate, and the resolution is generally greater than 20bp, but the DNA requirement is large and the resolution is low. When the fragment difference is less than 20bp, the result is unreliable

The resolution of polypropylene gel electrophoresis is better than that of agarose gel. 8%-10% polyacrylamide gel can detect PCR products with a difference of more than 10-20bp, but the operation is cumbersome, prone to gel leakage and uneven comb teeth and other phenomena

The interpretation of the results of the two traditional SSR detection methods is easily affected by uneven bands, blurred background, and tailing of swimming bands, resulting in unsatisfactory identification results and low analysis efficiency.

The resolution of fluorescent capillary electrophoresis is higher than that of traditional electrophoresis methods, but it is more expensive and prone to false positives. It can only judge the difference in product length and cannot detect the number of repeat units at SSR sites.

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