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Recombinant nucleic fragment RecCR010311 and detection primer and application thereof

A technology for recombining nucleic acids and fragments, which is applied in the field of molecular biology and can solve problems such as low efficiency and time-consuming

Active Publication Date: 2017-10-31
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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  • Recombinant nucleic fragment RecCR010311 and detection primer and application thereof
  • Recombinant nucleic fragment RecCR010311 and detection primer and application thereof
  • Recombinant nucleic fragment RecCR010311 and detection primer and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0075] Example 1 Breeding Recombinant Plants Introduced into Blast-resistant Genomic Fragments

[0076]The materials used in this example are rice 'Yuehui 9113' and rice 'Hua 3418B'.

[0077] The rice 'Hua 3418B' 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.

[0078] 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 desig...

Embodiment 2

[0088] Example 2 Determination of Homologous Recombination Fragments After Introduction of Rice Blast Resistance Genomic Fragments

[0089] In order to determine the size of the imported rice blast resistance genome fragment, homologous recombination fragments on both sides of the target genome fragment were sequenced for the homozygous single plant of the imported fragment of 'Yuehui 9113'. The recombinant nucleic acid fragment of the rice blast resistance genome contained in CR010311 was named RecCR010311.

[0090] It was preliminarily determined by the rice genome-wide breeding chip RICE60K detection results that RecCR010311 was located upstream of the SNP marker R0610434052AG.

[0091] At the same time, three samples of 'Yuehui 9113', 'Hua 3418B' and CR010311 were sequenced using Miseq sequencing technology. The TruSeq Nano DNALT Kit (illumina) kit was used for library establishment, the Library Quantification Kit–Universal (KAPA Biosystems) kit was used for quantificatio...

Embodiment 3

[0100] Example 3 Resistance Identification of 'Yuehui 9113' After Introducing the Rice Blast Resistance Genome Fragment

[0101] In order to identify the resistance effect, the new line CR010311 selected and bred by the present invention, the recurrent parent 'Yuehui 9113', 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) for indoor planting, it is cultivated to the 3-4 leaf stage and then identified by the following methods:

[0102] The 13-1209-1 strain isolated from rice blast samples in Guangdong disease nursery in 2013 was selected as the inoculation strain. 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 28°C for 5 days, take fresh ...

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Abstract

The invention relates to the molecular biology and specifically discloses a rice blast-resistant recombinant nucleic fragment RecCR010311 and a detection primer and application thereof. The invention further provides a method based on a whole genome selective breeding technology for selectively breeding rice plants comprising the rice blast-resistant genome recombinant nucleic fragment, a target genome segment is introduced into an acceptor material, and background recovery is achieved at the same time. A three line restorer Yuehui 9113 used widely is used as the improved acceptor material. Through the method, on the premise that the original characteristics of the Yuehui 9113 are reserved, the rice blast-resistant genome segment is introduced, and the rice blast resistance of the Yuehui 9113 is improved. Furthermore, the rice blast resistance of hybrids is greatly improved through combination. The genome recombinant nucleic fragment provided by the invention is closely related to the rice blast resistance and can be used as a resistance resource to be applied to cultivation of other varieties.

Description

technical field [0001] The invention relates to molecular biology, in particular to a rice blast-resistant recombined nucleic acid fragment. 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 to improve individual target traits, which can significantly shorten the breeding period. [0003] Rice blast is one of the most serious diseases of rice. The global rice yield loss caused by rice blast accounts for 11%-30% every year. Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11A01H1/02A01H1/04
CPCA01H1/02A01H1/04C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 喻辉辉周发松张学堂邱树青张小波雷昉姚玥潘丽李旭李菁韦懿陈光何予卿蒋建为张启发
Owner CHINA NAT SEED GRP
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