Rice genome recombinant nucleic acid fragment RecCR012612 and detection method thereof

A technology for recombinant nucleic acid and genome, applied in the field of recombinant nucleic acid fragments and its detection, can solve the problems of long time consumption and low efficiency, and achieve the effect of improving rice blast resistance

Active Publication Date: 2019-06-25
CHINA NAT SEED GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002]For a long time, the selection method of traditional breeding has mainly relied on the evaluation of field phenotypes, making choices based on the breeder’s personal experience. Low

Method used

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  • Rice genome recombinant nucleic acid fragment RecCR012612 and detection method thereof
  • Rice genome recombinant nucleic acid fragment RecCR012612 and detection method thereof
  • Rice genome recombinant nucleic acid fragment RecCR012612 and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Breeding of recombinant plants with gene fragments resistant to rice blast

[0043] The materials used in this embodiment are rice'Y58S' and rice'Gumei 4'.

[0044] Rice'Gumei 4'has good resistance to rice blast, and it is speculated that the Pi2 interval of chromosome 6 played a key role in the resistance of this material to rice blast.

[0045] In the process of selection of recombinant plants, molecular markers were used to perform prospect selection on recombinant plants, and the used prospect selection molecular markers were screened. Refer to the 6.1 edition of the MSU / TIGR annotation of the rice Nipponbare genome to download the 9,559,000 to 10,990,000 DNA sequence of chromosome 6. Use SSRLocator software to scan the SSR sites in the above sequence. Primer Premier 3.0 software was used to design primers for the found SSR sites, and a total of 162 pairs of primers were designed. The PCR method was used to screen the polymorphisms of the above primer pairs i...

Embodiment 2

[0055] Example 2 Determination of homologous recombination fragments after introduction of rice blast resistance genome fragments

[0056] In order to determine the size of the introduced rice blast resistance genome fragment, the homozygous single plant of the'Y58S' introduced fragment was sequenced for the homologous recombination fragments on both sides of the target genome fragment. The recombinant nucleic acid fragment of the rice blast resistant genome contained in CR012612 was named RecCR012612.

[0057] Preliminarily confirmed by the detection results of the rice whole genome breeding chip RICE60K, the upstream homologous recombination fragment of RecCR012612 is located between the markers R0610187834CT and F0610202034AC, and the downstream homologous recombination fragment is located between the markers F0610692034AG and R0610699627AG.

[0058] At the same time, the Miseq sequencing technology was used to perform whole-genome sequencing on three samples of'Y58S','Gumei No. ...

Embodiment 3

[0071] Example 3 Resistance identification of ‘Y58S’ after introducing the rice blast resistance genome fragment

[0072] In order to identify the resistance effect, the new line CR012612, the recurrent parent'Y58S', the rice blast resistant variety Gumei 4 (as a positive control), and the rice blast susceptible variety Lijiang Xintuan Heigu (as a negative Control) was planted indoors, and then cultivated to the 3-4 leaf stage for identification using the following methods:

[0073] Select M15Bb-1-1, M15Bb-1-2, M15Bb-2-1, M15Bb-3-1, M15Bb-4-1, M15Bb-5-1, M15Bb-6-1 separated from Yichang in 2015. 7 Two strains were used as inoculation strains. Strains are stored at -20°C using the sorghum grain method. Before use, remove the stored sorghum grains to a potato dextrose medium (PDA) plate for activation (PDA: peeled potatoes 200g, glucose 20g, agar powder 15g, distilled water to make the volume to 1L), After 5 days of light cultivation at 28°C, take a fresh mycelial block with a di...

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Abstract

The invention provides a rice genome recombinant nucleic acid fragment RecCR012612 and a detection method thereof. The present invention also provides a method for seed selection of a rice plant containing a recombinant nucleic acid fragment, a molecular marker is used to perform foreground selection and background selection on the recombinant plant, and the rice plant containing the recombinant nucleic acid fragment is obtained.

Description

Technical field [0001] This application relates to genome-wide selective breeding technology. Specifically, this application relates to the use of whole-genome selective breeding technology to select rice plants with recombinant nucleic acid fragments resistant to rice blast, as well as the recombinant nucleic acid fragments obtained therefrom and detection methods thereof. Background technique [0002] For a long time, the selection method of traditional breeding mainly relied on the evaluation of field phenotype, and the choice was made based on the personal experience of the breeder. The biggest disadvantage is that it is time-consuming and low in efficiency. To improve the efficiency of selection, the most ideal method should be able to directly select genotypes. With the development of molecular biotechnology, molecular markers provide the possibility for direct selection of genotypes. In recent years, molecular marker-assisted selection methods have been used to improve i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11A01H1/02A01H1/04
CPCY02A40/146
Inventor 周发松邱树青曹志喻辉辉张学堂张龙雨雷昉姚玥李旭江峥李菁韦懿何予卿张启发
Owner CHINA NAT SEED GRP
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