Molecular SNP markers linked to qpsiib10, the main QTL locus for peanut resistance to Aspergillus flavus infection, and their applications

A molecular marker, the technology of Aspergillus flavus, which is applied in the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of the small number of major QTLs, the application of difficult-to-flower breeding varieties, etc., to improve the breeding efficiency Effect

Active Publication Date: 2022-04-08
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0005] At present, there are few studies on the QTL mapping of peanut kernel resistance to Aspergillus flavus infection, and the number of main QTLs detected is relatively small, which is difficult to apply in peanut breeding

Method used

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  • Molecular SNP markers linked to qpsiib10, the main QTL locus for peanut resistance to Aspergillus flavus infection, and their applications
  • Molecular SNP markers linked to qpsiib10, the main QTL locus for peanut resistance to Aspergillus flavus infection, and their applications
  • Molecular SNP markers linked to qpsiib10, the main QTL locus for peanut resistance to Aspergillus flavus infection, and their applications

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Test materials: The peanut variety Zhonghua 16 was used as the female parent, and the peanut germplasm J11 was used as the male parent for crossing, and a recombinant inbred line (RIL) population F containing 200 lines was obtained by single seed transmission method 7(2017), F 8 (2018) and F 9 (2019).

[0041] Phenotype identification: Planting Zhonghua 16, J11 and F 7 (2017), F 8 (2018) and F 9 (2019) Generation of 200 strains from the RIL population. A completely randomized block experimental design was adopted, with 3 repetitions. Plant 1 row of 10-12 plants per material for each repetition, with a row spacing of 30 cm and a plant spacing of 10 cm. Adopt standard field management practices. When ripe, healthy plants are selected to harvest pods, dried and stored properly. After shelling, 20 typical seed kernels were selected to inoculate Aspergillus flavus indoors, and the infection level of each seed was recorded after the onset was sufficient: Level 0, no A...

Embodiment 2

[0047] The 200 RILs of Zhonghua 16 and J11 (see Example 1) were detected using two SNP markers linked to the main locus qPSIIB10 for resistance to Aspergillus flavus infection. Using genomic DNA as a template and the sequences shown in SEQ ID NO.3-4 and SEQ ID NO.5-6 as primers, two molecular markers were simultaneously amplified using the KAPA2G Fast Multiplex Mix kit. PCR conditions were: 95°C pre-denaturation for 3 minutes; 95°C denaturation for 15 s, 55°C renaturation for 30 s, 72°C extension for 30 s, a total of 30 cycles; finally 72°C extension for 5 min, 4°C incubation. After the amplification product was added with a sequencing adapter, the Illumina HiSeq platform was used for Paired-end 150bp (PE150) sequencing, and the sequencing sequence was compared to the reference sequence to detect the base of the SNP site. If the bases of the RIL and the SNP sites detected in the amplified fragment of Zhonghua 16 are consistent, it indicates that the strain contains the allele ...

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Abstract

The invention relates to the technical field of molecular markers, in particular to a SNP molecular marker linked with the main effect QTL site qPSIIB10 of peanut resistance to Aspergillus flavus infection and its application. The SNP molecular markers of the present invention linked to the main effect QTL loci for peanut resistance to Aspergillus flavus infection include: qPSIIB10-L or qPSIIB10-R. Wherein, the SNP molecular marker qPSIIB10‑L contains a nucleotide sequence in which the 235th polymorphism of the sequence shown in SEQ ID NO.1 is C / T and the 338th polymorphism is G / A; the SNP molecular marker qPSIIB10‑R contains a nucleotide sequence whose polymorphism is G / A at position 297 of the sequence shown in SEQ ID NO.2. The invention can assist in selecting materials for peanut kernels resistant to aspergillus flavus infection, and is beneficial to promoting the selection and application of new varieties of peanuts resistant to aspergillus flavus.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to a SNP molecular marker linked to the main effect QTL site qPSIIB10 of peanut resistance to Aspergillus flavus infection and its application. Background technique [0002] Peanut (Arachis hypogea L.) is one of the important oil crops. Peanut seeds are rich in edible oils, proteins, amino acids and vitamins, and can be processed into edible oils, nuts, peanut butter and other foods. However, during peanut production and after harvesting, peanuts may be infected by Aspergillus flavus, which produces aflatoxin and seriously damages the quality of peanuts. Therefore, how to reduce the infection of Aspergillus flavus on peanut seeds and control the harm of aflatoxin is a major demand for the development of peanut industry. [0003] Breeding and utilizing disease-resistant varieties is one of the most cost-effective measures to control crop diseases. The study found that so...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 廖伯寿罗怀勇雷永黄莉刘念晏立英康彦平姜慧芳蒋艺飞
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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