Construction method of corylus plant variety SSR molecular marker fingerprint spectrum

Abstract

The invention discloses a construction method of a corylus plant variety SSR molecular marker fingerprint spectrum. The construction method comprises the following steps of: (1) extracting the corylusplant DNA of a variety which participates in selection; (2) using 25 pairs of core SSR molecular marker primers to independently carry out multiple PCR amplification on a variety to be tested; (3) grouping an amplification product in (2), and then, carrying out mixing according to groups to carry out capillary electrophoresis detection; and (4) carrying out data analysis on a detection result, and constructing the fingerprint spectrum. In the construction method, a core primer is high in polymorphism, data can be easily read, each species of a corylus plant is compatible, and a distinguishingfunction is powerful. Through a primer mixing PCR technology, one-time PCR can obtain the products of two groups of primers, and one-multiple PCR cost is saved. Through a PCR product mixing capillaryelectrophoresis technology, the data information of four to eight groups of primers can be simultaneously obtained through one-time electrophoresis, three-to-seven-multiple capillary electrophoresisexpense is saved, and therefore, the purposes of saving cost for an experiment operation and being economic and quick is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for constructing SSR minute marker fingerprints of Corylus species. Background technique [0002] Corylus (Corylus) belongs to the Betuceae (Betulaceae) Hazel family. There are 13 wild species widely recognized in the world. The widely cultivated hazelnut species are European Hazel (C.avellana L.) (C.maximaMill.), the main varieties are: 'Tombul' (Turkey), 'TGDL' (Italy), 'Barcelona' (USA), 'Jefferson' (USA), etc. [0003] There are 8 species and 2 varieties of Corylus native to China, more than 90% of which are wild flat hazel (C.heterophylla Fisch.). The main cultivation is Pingou hybrid hazel and a small amount of horticultural cultivation of flat hazel. Pingou hybrid hazel (Corylusheterophylla Fisch.×Corylusavellana L.) is a hybrid offspring obtained by domestic experts in the 1980s by crossing Chinese Pine hazel and foreign hazel species. Up to now, in...

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Problems solved by technology

[0012] (1) RAPD, ISSR and AFLP markers are all dominant markers, the number of amplified fragments is large, and false positive fragments are prone to exist, resulting in reduced identification accuracy;

[0013] (2) The difference between samples is distinguished by the relative position of the amplified bands and the clustering diagram generated, and the size of the bands is not digitized;

[0014] (3) Only a limited number of Pingou hybrid hazel varieties (strains) are involved, and they do not have the versatility of Corylus plant varieties;

[0015] (4) Variety identification methods are different in different laboratories and different batches of experiments, or rely on control varieties, the method is not universal and convenient

[0017] (1) The detection scope only includes a limited number of Pingou hybrid hazel varieties (strains), and does not involve other hazel varieties;

[0018] (2) The source and test range of the primers are Pingou hybrid hazelnuts, and there is uncertainty in the universality of all Corylus species at home and abroad;

[0019] (3) The PIC value of the primers is low, and there is uncertainty in the distinction of similar varieties;

[0020] (4) The cost of PCR and capillary electrophoresis is relatively high in the test operation

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