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Primer sequence for authenticating resistance of strawberry genetic resource anthracnose and authentication method thereof

A technology for anthracnose and strawberry, applied in the field of primer sequence and identification for identification of anthracnose resistance of strawberry germplasm resources, can solve the problems of quality trait inheritance, high-level single gene control of resistance, etc.

Active Publication Date: 2014-03-26
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

Denoyes-Rothan et al. (2005) designed an 8×8 factorial experiment to study the genetic characteristics of strawberry anthracnose C.acutatum species, and believed that although minor polygenes can affect the resistance level of strawberry, high-level resistance is still considered a single gene Control, which belongs to the inheritance of quality traits

Method used

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  • Primer sequence for authenticating resistance of strawberry genetic resource anthracnose and authentication method thereof
  • Primer sequence for authenticating resistance of strawberry genetic resource anthracnose and authentication method thereof
  • Primer sequence for authenticating resistance of strawberry genetic resource anthracnose and authentication method thereof

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Embodiment 1

[0054] Using self-designed 210 pairs of SSR primers and AFLP primers (see Table 1), a total of 245 DNA samples from hybrid parents (Daselect and Sweet Charlie), hybrid F1 and F2 inbred lines (see Table 2) Screening and identification of strawberry anthracnose resistance by AP-PCR molecular markers.

[0055] 1. DNA extraction

[0056] To identify a single strawberry plant (No. 6 single plant, the F1 inbred line of Daselect and Sweet Charlie hybrid), cut the 1.5 cm long leaves when sampling. Use the modified CTAB extraction solution (2%CTAB, 2%PVP, pH=8.0100mM Tris-Cl, pH=8.050mM EDTA, 1.4M KCl, 2%BME) to quickly extract DNA. The specific operation process is as follows:

[0057] ①Pre-cool the mortar with liquid nitrogen, quickly take an appropriate amount of strawberry leaves at -70°C and store them in the mortar, add liquid nitrogen to fully grind them into powder, transfer to a 2.0ml centrifuge tube pre-filled with 1.4ml STE nuclear separation solution, mix them upside down,...

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Abstract

The invention discloses a primer sequence for authenticating resistance of strawberry genetic resource anthracnose and an authentication method thereof. The method is characterized in that the genome DNA of 250 parts of filial generation materials obtained through the conventional cross breeding by adopting sweet Charlie and Darselect as cross parents as a template, the PCR amplification is executed for 146 pairs of SSR primers and 64 pairs of AFLP primers covering the complete genome, the specificity genetic map of the resistance of strawberry anthracnose is established through the PAGE microgel electrophoresis analysis, 10 pairs of SSR primers with high specificity and strong dominance of the filial generation are successfully obtained, and the nucleotide sequence of the SSR primer is indicated as SEQ ID No.1 to 20. When the primer sequence obtained through the method is used for authenticating the resistance of the strawberry genetic resource anthracnose, the matching rate of an authentication result and the field resistance can reach 88.4 percent. The method has the advantages of simplicity, rapidness, high efficiency, high breeding efficiency, short breeding period and the like and has high application value on the authentication of the resistance of the strawberry genetic resource anthracnose.

Description

technical field [0001] The invention relates to a primer sequence for identifying anthracnose resistance of strawberry germplasm resources and an identification method thereof. Background technique [0002] Strawberry anthracnose caused by the anthracnose fungus (Colletotrichumn spp.) is a devastating disease in the world. Up to now, there are still large-scale harms of the disease on strawberry production in many countries. Anthracnose can harm strawberry stems, petioles, leaves, fruits, flowers, flower buds, sepals and other organs, not only can cause serious loss of economic output, but also some diseased residues contain more mycotoxins, which seriously threaten people. food safety. [0003] Strawberry anthracnose was first discovered in France in 1832, 180 years ago. There have been some reports on the identification of resistance to strawberry anthracnose pathogens. The identification of stress resistance of germplasm resources is very important for fruit scientific...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 高清华叶正文段可郑宏清李静刘建成傅弘平
Owner SHANGHAI ACAD OF AGRI SCI
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