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Indel Molecular Marker Closely Linked with Rice Heading Time Gene and Its Application

A technology of molecular markers and heading date, which is applied in the field of DNA molecular marker technology and molecular biology, and can solve the problems of InDel marker unsaturation

Active Publication Date: 2022-06-03
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 Rice Heading Time Gene and Its Application
  • Indel Molecular Marker Closely Linked with Rice Heading Time Gene and Its Application
  • Indel Molecular Marker Closely Linked with Rice Heading Time Gene and Its Application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Design of InDel Molecular Marker Detection Primers Closely Linked to QTLs at Heading Stage of Rice

[0058] 1.1 Primer design method:

[0059] In the chromosome segments that need to develop InDel markers, molecular markers were designed based on the results of japonica (Nipponbare) and indica (9311) genome sequencing. 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). The desired PAC / BAC clones were selected and each clone was divided into 2kb contiguous small fragments. Then, these 2kb sequences were used for sequence alignment with indica rice variety 9311, and the sequences with insertion / deletion bases of 7-12bp were selected as candidate sequences. Based on the Nipponbare genome sequence, primers were designed within 200bp of each flank of the screened InDel site. Primers were designed so that Primer3plus ( http: / / www.bioinformatics.nl / cgi-bin / prim...

Embodiment 2

[0090] Primer Application of InDel Molecular Markers IN2 and IN10 in Rice Gene

[0091] 2.1 Rice genomic DNA extraction

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

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

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

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

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

[0097] 2.1.4. Add pre-cooled isopropanol or anh...

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Abstract

The invention discloses an InDel molecular marker closely linked with a rice heading stage gene and its application. The upstream primer sequence of the molecular marker IN2 is SEQ ID NO.2, and the downstream primer sequence is SEQ ID NO.3; the InDel molecular marker IN2 is used in rice Application in genetic analysis of heading stage genes and gene fine positioning; the upstream primer sequence of the molecular marker IN10 is SEQ ID NO.6, and the downstream primer sequence is SEQ ID NO.7; the InDel molecular marker IN10 is used in rice heading stage gene Applications in genetic analysis and fine mapping of genes.

Description

technical field [0001] The invention relates to an InDel molecular marker closely linked with rice heading stage genes and its application, in particular to an InDel (insertion-deletion polymorphisms) molecular marker and its application in genetic analysis and fine mapping of rice heading stage genes, belonging to DNA molecular marker technology and The field of molecular biology technology. Background technique [0002] Insertion / deletion polymorphism (insertion-deletion, InDel) molecular markers are molecular markers developed according to the insertion or deletion of nucleotide fragments in homologous sequences in different individual genomes, that is, relative to one genotype, another gene There are a certain number of nucleotide insertions or deletions in the genome of the type. InDel markers have both the characteristics of simple sequence repeats (SSRs) and single nucleotide polymorphisms (Single nucleotide polymorphisms, SNPs) variation, and are a class of codomina...

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 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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