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Rice genome recombinant nucleic acid fragment RecCR012613 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

AI Technical Summary

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 RecCR012613 and detection method thereof
  • Rice genome recombinant nucleic acid fragment RecCR012613 and detection method thereof
  • Rice genome recombinant nucleic acid fragment RecCR012613 and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1Breeding of Recombinant Plants Introduced with Blast Resistance Genome Fragment

[0043] The materials used in this example are rice 'Y58S' and rice 'Gumei No. 4'.

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

[0045] During the breeding process of the recombinant plants, the molecular markers were used to perform prospect selection on the recombinant plants, and the adopted molecular markers for prospect selection were screened. Referring to the rice Nipponbare genome MSU / TIGR annotation version 6.1, download the DNA sequence of chromosome 6 from 9,559,000 to 10,990,000. The SSR sites in the above sequences were scanned using SSRLocator software. Primer Premier 3.0 software was used to design primers for the found SSR loci, and a total of 162 pairs of primers were designed. By means of PCR, the polymorphisms of th...

Embodiment 2

[0055] Example 2 Determination of Homologous Recombination Fragments After Introducing Blast Resistance Genome Fragments

[0056] In order to determine the size of the imported rice blast resistance genome fragment, the homozygous individual plants of the 'Y58S' imported fragment were sequenced for homologous recombination fragments on both sides of the target genome fragment. The recombinant nucleic acid fragment of the rice blast resistance genome contained in CR012613 was named RecCR012613.

[0057] It was preliminarily determined by the rice genome-wide breeding chip RICE60K detection results that the upstream homologous recombination fragment of RecCR012613 was located between the markers R0610178008AC and R0610183178CT, and the downstream homologous recombination fragment was located between the markers F0610739800TC and R0610747083GT.

[0058] At the same time, three samples of 'Y58S', 'Gumei 4' and CR012613 were sequenced whole genomes using Miseq sequencing technolo...

Embodiment 3

[0072] Example 3 Resistance Identification of 'Y58S' After Introducing Blast Resistance Genome Fragment

[0073] In order to identify the resistance effect, the new line CR012613 selected by the application, the recurrent parent 'Y58S', the rice blast resistant variety Gumei No. 4 (as a positive control), and the rice blast susceptible variety Lijiang Xintuan Heigu (as a negative control) Control) is carried out indoor planting, and adopts following method to identify after it is cultivated to the 3-4 leaf stage:

[0074]A total of 7 M15Bb-1-1, M15Bb-1-2, M15Bb-2-1, M15Bb-3-1, M15Bb-4-1, M15Bb-5-1, M15Bb-6-1 isolated from Yichang in 2015 were selected. strains were used as inoculation strains. The strain was stored at -20°C by the sorghum grain method. Before use, the preserved sorghum grains were taken out and activated on a potato dextrose medium (PDA) plate (PDA: 200g peeled potatoes, 20g glucose, 15g agar powder, distilled water to 1L), After cultivating under light at...

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Abstract

The invention provides a rice genome recombinant nucleic acid fragment RecCR012613 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, the application relates to the selection and breeding of rice plants with recombinant nucleic acid fragments with rice blast resistance function by using genome-wide selective breeding technology, as well as the recombinant nucleic acid fragments obtained thereby and detection methods thereof. Background technique [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. The biggest disadvantage is that it takes a long time and is low in efficiency. To improve the efficiency of selection, the most ideal method should be to be able to directly select the genotype. With the development of molecular biotechnology, molecular markers provide the possibility for direct selection of genotypes. In recent years, molecular marker-assisted selection m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11A01H1/02A01H1/04
Inventor 李旭周发松曹志喻辉辉邱树青张学堂张小波雷昉姚玥江峥李菁韦懿何予卿张启发
Owner CHINA NAT SEED GRP
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