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InDel molecular marker closely linked with gen of rice in heading stage and application

A molecular marker and heading stage technology, applied in the field of DNA molecular marker technology and molecular biology, can solve the problem of unsaturated InDel marker, and achieve the effect of obvious band type and polymorphism

Active Publication Date: 2020-06-19
BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a large number of InDel markers have been developed according to the results of rice genome sequencing, but the development of rice InDel markers is still not saturated

Method used

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  • InDel molecular marker closely linked with gen of rice in heading stage and application
  • InDel molecular marker closely linked with gen of rice in heading stage and application
  • InDel molecular marker closely linked with gen of rice in heading stage and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Design of InDel Molecular Marker Detection Primers Tightly Linked to Heading Date QTL in Rice

[0058] 1.1 Primer design method:

[0059] In the chromosomal segments where InDel markers need to be developed, molecular markers were designed based on the genome sequencing results of japonica rice (Nipponbare) and indica rice (9311). The sequence of the Nipponbare PAC / BAC clone in this region was downloaded from the TIGR website (The Institute of Genome Research) (http: / / www.tigr.org / tdb / rice). Select the required PAC / BAC clones, and split each clone into 2kb contiguous small fragments. Then use these 2kb sequences to compare the sequences with the indica rice variety 9311, and screen the sequences with insertion / deletion bases of 7-12 bp as candidate sequences. Based on the Nipponbare genome sequence, primers were designed within 200 bp of each flank of the screened InDel site. The primers were designed so that Primer3plus ( http: / / www.bioinformatics.nl / cgi-bin / prime...

Embodiment 2

[0090] Application of Primers for InDel Molecular Markers IN2 and IN10 in Rice Genes

[0091] 2.1 Rice genomic DNA extraction

[0092] CSSL41, Shengdao 16, and 24 exchange lines (A1-A24) obtained by screening all F2 generation recessive (late heading) populations with RM527 and PSM388 as boundary markers.

[0093] The total DNA of rice genome was extracted by the improved TPS simple method, and the specific steps were as follows:

[0094] 2.1.1. At the peak tillering stage, take 1 to 2 young leaves from the upper part of each plant, and store them in a -80°C refrigerator for later use;

[0095] 2.1.2. When extracting DNA, take 2-4cm long rice leaves and put them into a 1.5ml centrifuge tube, put them in liquid nitrogen, grind them, add 900ml of TPS extract solution, and bathe in 75°C water bath for 30-60min;

[0096] 2.1.3 Centrifuge at 12000rpm for 10min, absorb about 500ml of the supernatant and transfer to a new 1.5ml centrifuge tube;

[0097] 2.1.4. Add pre-cooled isopr...

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Abstract

The invention discloses an InDel molecular marker closely linked with a gene of rice in a heading stage and an application. The molecular marker IN2 has an upstream primer sequence of SEQ ID NO. 2 anda downstream primer sequence of SEQ ID NO. 3. The invention discloses the application of the InDel molecular marker IN2 in genetic analysis and fine gene mapping of the rice in the heading period. The molecular marker IN10 has an upstream primer sequence of SEQ ID NO. 6 and a downstream primer sequence of SEQ ID NO. 7. The invention discloses the application of the InDel molecular marker IN10 ingene genetic analysis and gene fine mapping of the rice in the heading stage.

Description

technical field [0001] The present invention relates to the InDel molecular marker closely linked with the rice heading stage gene and its application, in particular to the InDel (insertion-deletion polymorphisms) molecular marker and its application in genetic analysis and fine positioning of the rice heading stage gene, belonging to DNA molecular marker technology and field of molecular biology. Background technique [0002] Insertion-deletion polymorphism (insertion-deletion, InDel) molecular marker is a molecular marker developed based on the insertion or deletion of nucleotide fragments in the homologous sequence of the genome of different individuals, that is, relative to one of the genotypes, another gene There is a certain number of nucleotide insertions or deletions in the genome of the type. InDel markers combine the characteristics of simple sequence repeats (Simple sequence repeats, SSRs) and single nucleotide polymorphisms (Single nucleotide polymorphisms, SNPs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 宣宁柳絮张华姚方印赵倩倩
Owner BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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