Recombinant nucleic fragment RecCR010380 and detection method thereof

A technology for recombining nucleic acids and fragments, applied in the field of recombinant nucleic acid fragments and their detection, can solve the problems of long time and low efficiency, and achieve the effects of improving rice blast resistance and improving rice blast resistance.

Active Publication Date: 2017-03-08
CHINA NAT SEED GRP
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  • 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 e

Method used

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  • Recombinant nucleic fragment RecCR010380 and detection method thereof
  • Recombinant nucleic fragment RecCR010380 and detection method thereof
  • Recombinant nucleic fragment RecCR010380 and detection method thereof

Examples

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Effect test

Embodiment 1

[0040] Example 1 Breeding of Recombinant Plants Introduced with Blast Resistance Genome Fragment

[0041] The materials used in this example are rice 'Zhongzhonghui 629' and rice 'Hua 130B'.

[0042] The rice 'Hua 130B' has good resistance to rice blast, and it is speculated that the gene cluster region where Pi2, Pi9 and Pigm of chromosome 6 are located may play a key role in the rice blast resistance of this material.

[0043] 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 w...

Embodiment 2

[0053] Example 2 Determination of Homologous Recombination Fragments After Introducing Blast Resistance Genome Fragments

[0054] In order to determine the size of the imported rice blast resistance genome fragment, the homozygous individual plants of the imported fragment of 'Zhongzhonghui 629' 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 CR010380 was named RecCR010380.

[0055] It was preliminarily determined by the rice genome-wide breeding chip RICE60K detection results that RecCR010380 was located between two SNP markers R0610178008AC and R0610420370GA.

[0056] At the same time, three samples of 'Zhongzhonghui 629', 'Hua 130B' and CR010380 were subjected to whole-genome sequencing using Miseq sequencing technology. The TruSeq Nano DNA LT Kit (illumina) kit was used for library establishment, the Library Quantification Kit–Univers...

Embodiment 3

[0066] Example 3 Identification of rice blast resistance after introduction of 'Zhongzhonghui 629' genome fragments

[0067] In order to identify the resistance effect, the new strain CR010380 selected by the application, the reincarnation parent 'Zhongzhonghui 629', the rice blast resistant variety Gumei No. 4 (as a positive control), and the rice blast susceptible variety Lijiang Xintuan Heigu (as negative control) carry out indoor planting, adopt following method to identify after it is cultivated to 3-4 leaf stage:

[0068] Select 6102-1 isolated from rice blast diseased leaves and necks in Sichuan disease garden in 2013, 21K14-1, 21K02-1, 4105-1 in Hubei disease garden, 1209-1 in Guangdong disease garden, and Fujian disease garden 8111-1, a total of 6 rice blast 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 ...

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Abstract

The application provides a recombinant nucleic fragment and a detection method thereof and also provides a breeding method of a rice plant with the recombinant nucleic fragment; the rice plant with the recombinant nucleic fragment is acquired by using a molecular marker to perform foreground selection and background selection on recombinant plants.

Description

technical field [0001] This application relates to genome-wide selective breeding technology. Specifically, the present application relates to the selection and breeding of rice plants containing recombinant nucleic acid fragments using the genome-wide selective breeding technology, as well as the recombinant nucleic acid fragments obtained therefrom and their detection methods. 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 methods have been used...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12Q1/68C12N15/11A01H1/02A01H1/04
Inventor 周发松喻辉辉张学堂邱树青张小波雷昉律文堂姚玥潘丽李旭李菁韦懿陈光何予卿杨毅田冰川张启发
Owner CHINA NAT SEED GRP
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