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Molecular markers and applications of ssr tightly linked to rice purple leaf sheath qtl

A purple leaf sheath and molecular marker technology, applied in the field of DNA molecular marker technology and molecular biology, can solve the problem of lack of saturation in the development of rice SSR markers

Inactive Publication Date: 2019-03-15
BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, a large number of SSR markers have been developed based on the rice genome sequencing results (McCouch et al., 2002; Saini et al., 2004; Nonoue et al., 2008; Matsubara et al., 2008; Maas et al., 2010), However, the development of rice SSR markers is still not saturated

Method used

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  • Molecular markers and applications of ssr tightly linked to rice purple leaf sheath qtl
  • Molecular markers and applications of ssr tightly linked to rice purple leaf sheath qtl
  • Molecular markers and applications of ssr tightly linked to rice purple leaf sheath qtl

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Experimental program
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Embodiment 1

[0031] The design of embodiment 1 rice SSR marker primer

[0032] 1.1 Primer design method:

[0033]Molecular marker design is carried out on the chromosome segments where microsatellite markers need to be developed. The sequences of PAC / BAC clones in this region were downloaded from the TIGR website (The Institute of Genome Research) (http: / / www.tigr.org / tdb / rice). TIGR has integrated PAC / BAC clones sequenced by IRGSP (International Rice Genome Sequencing Project, IRGSP) into the genetic map. The sequences of PAC / BAC clones downloaded from the target region first need to use the repetitive sequence search software SSRIT (http: / / www .gramene.org / microsat / ) to search the microsatellite sequence, and then select the appropriate microsatellite sequence (generally require the number of repeats to be more than 8, and avoid the motif being AT), and download a small segment containing the microsatellite sequence Genome sequence (350-500bp), and finally use this sequence to design P...

Embodiment 2

[0055] Example 2 Application of Rice SSR Molecular Marker PSH10, SSR Molecular Marker PSH13, and SSR Molecular Marker PSH15

[0056] 2.1 Rice genomic DNA extraction

[0057] The total DNA of rice genome was extracted by the improved TPS simple method, and the specific steps were as follows:

[0058] 1. At the peak tillering stage, take 1-2 young leaves from the upper part of each plant, and store them in a -80°C refrigerator for later use;

[0059] 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;

[0060] 3. Centrifuge at 12000rpm for 10min, absorb about 500ml of the supernatant and transfer to a new 1.5ml centrifuge tube;

[0061] 4. Add pre-cooled isopropanol or absolute ethanol to top up. Overnight at 4°C, centrifuge at 12000rpm for 10min;

[0062] 5. Discard the supernatant, dry the precipitate, add 150 μl ...

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Abstract

The invention relates to an SSR (Simple Sequence Repeats) molecular marker tightly linked with rice purple leaf sheath QTL (Quantitative Trait Locus) and application. The nucleotide sequence of an upstream marker of the SSR molecular marker PSH10 tightly linked with rice purple leaf sheath QTL is shown as SEQ ID NO.1, and the nucleotide sequence of a downstream marker of the SSR molecular marker PSH10 tightly linked with rice purple leaf sheath QTL is shown as SEQ ID NO.2. The SSR molecular marker PSH13 tightly linked with rice purple leaf sheath QTL and the SSR molecular marker PSH15 tightly linked with rice purple leaf sheath QTL are also provided. The three pairs of SSR molecular markers PSH10, PSH13 and PSH15 tightly linked with rice purple leaf sheath QTL are screened out for the first time. The rice purple leaf sheath genes can realize genetic analysis and precise positioning by using the three groups of SSR molecular markers, and the three groups of SSR molecular markers can be used for building a rice high-density genetic map and a variety fingerprint map.

Description

technical field [0001] The present invention relates to the SSR molecular marker closely linked with the rice purple leaf sheath QTL and its application, in particular to the rice simple repeat sequence (SSR, Simple sequence repeats) molecular marker and its application in the genetic analysis and fine positioning of the rice purple leaf sheath gene. DNA molecular marker technology and molecular biology technology field. Background technique [0002] Molecular markers are currently widely used in the analysis of genetic diversity of germplasm resources and assisted breeding, and are an important and indispensable tool for breeding and genetics research. Especially the emergence of SSR (Akkaya et al., 1992), RAPD (Williams et al., 1990), etc., because of their simplicity, high efficiency and rapidity, the development and utilization of molecular markers have spread to various crops. [0003] SSR (simple sequence repeat, simple repeat sequence), also known as microsatellites ...

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 姚方印陈高柳絮张华宣宁杨永义
Owner BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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