Peanut oligonucleotide probes and their design method and use method

An oligonucleotide probe and oligonucleotide technology are applied in the field of peanut oligonucleotide probes and their design, which can solve the problems of difficult development and identification of peanut cytology markers, and achieve economical efficiency, versatility, The effect of good application prospects

Active Publication Date: 2017-07-28
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is to overcome the difficulty in developing peanut cytological markers, design a cost-effective and versatile peanut oligonucleotide probe, and increase peanut cytological markers; and apply the newly designed oligonucleotides Acid probes establish fluorescent in sit

Method used

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  • Peanut oligonucleotide probes and their design method and use method
  • Peanut oligonucleotide probes and their design method and use method

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Embodiment 1

[0060] Embodiment 1, a kind of design method of peanut oligonucleotide probe

[0061] The design method of peanut oligonucleotide probe of the present invention comprises following two modes:

[0062] (a) Development of oligonucleotide probes using microsatellite (SSR) and telomeric sequences

[0063] All ten forms of three-base SSR oligonucleotides (AAC)10, (AAG)10, (AGG)10, (ACT)10, (CAT)10, (CAC)10, (ACG)10, (CAG)10, (ATT)10 and (GCC)10, and then using fluorescence in situ hybridization to screen the root tip metaphase chromosomes of the peanut cultivar "Sizihong", it was found that the probes Oligo DP-2 and Oligo DP-4 were in the four There is a signal on the red chromosome, further use 45S rDNA, 5S rDNA, A.duranensis and Genomic DNA sequence FISH analysis of newly developed oligonucleotide probes OligoDP-2 and Oligo DP-4 in A. duranensis and A. duranensis location on the chromosome. OligoDP-2 and Oligo DP-4 were found in four grains of red, A.duranensis and Stable ...

Embodiment 2

[0076] Embodiment 2, the using method of peanut oligonucleotide probe

[0077] 1. Design of probe cover

[0078] The present invention utilizes Oligo DP-1~Oligo DP-7 to assemble two groups of probe sets, respectively Oligo#1 (FAM marked Oligo DP-2, Oligo DP-5 and Oligo DP-7) and Oligo#2 ( TAMARA-labeled Oligo DP-1, OligoDP-3, Oligo DP-4 and Oligo DP-6), the probe concentration is shown in Table 1.

[0079] 2. Test materials

[0080] The peanut cultivar Sizihong and 8 diploid wild species were preserved by the Economic Crops Research Institute of Henan Academy of Agricultural Sciences (Table 2).

[0081] Table 1: Peanut Oligonucleotide Probe Sets

[0082]

[0083] Table 2: Blocks and codes of Sizihong and 8 wild peanut species

[0084]

[0085] 3. Fluorescence in situ hybridization mixed staining analysis

[0086] The probe set was used to analyze the mixed staining of peanut cultivar Sizihong and 8 diploid wild species, including the following steps:

[0087] (1) P...

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Abstract

The invention discloses peanut oligonucleotide probes and their design method and use method. The peanut oligonucleotide probes comprise eight probes having nucleotide sequences shown in the formulas of SEQ ID NO. 1 to NO. 8. The microsatellite and telomere sequences are used for developing the oligonucleotide probes, the novel oligonucleotide probes are combined into a probe set and oligonucleotide types of the peanut cultivar and wild peanut are constructed, an economic, efficient and high universality peanut cytological marker design technique and chromosome recognition technique are built, peanut chromosome markers are enriched, genomes and chromosomes of the peanut cultivar and wild peanut are identified, and chromosomal structural variation of wild peanut species is identified. Through use of a high-throughput small data simplified sequencing (for only measuring the amount of genomic 4Gb data) and bioinformatics analysis, peanut oligonucleotide probe markers are successfully developed. The invention provides a novel effective method for low-cost and high-efficiency development of peanut cytological markers.

Description

technical field [0001] The invention relates to the design and usage of a peanut cytological marker, in particular to a peanut oligonucleotide probe and its design method and usage method. Background technique [0002] Peanut is an important oil crop and economic crop in the world, and it is an important source of vegetable oil and protein in many developing countries. China is the world's largest peanut producer, consumer and exporter, with a total peanut production of about 16.5 million tons in 2016. [0003] So far, about 80 species of the genus Arachis have been named and identified. According to the characteristics of hybrid affinity and morphology between the species of the genus Arachis, the genus Arachis is divided into 9 sections, including the Arachis section ( A, B and D Genomes), Peripheral Blocks (Ex), Large Root Blocks (C), Three-Particle Blocks (Tris), Prostrate Blocks (P), Heteromorphic Floral Blocks (H), Erect Blocks ( E), trefoil block (Trie) and rhizome ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6841C12Q1/6895C12Q2600/13C12Q2600/156C12Q2563/107C12Q2543/10
Inventor 张新友杜培亓增军李丽娜董文召黄冰艳汤丰收韩锁义
Owner HENAN ACAD OF AGRI SCI
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