Recombinant nucleic fragment RecCR010005 and detection primer and application thereof

A technology for recombining nucleic acids and fragments, applied in the field of molecular biology, can solve the problems of low efficiency and long time, and achieve the effect of improving rice blast resistance and improving rice blast resistance.

Active Publication Date: 2017-10-31
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 evaluation of field phenotypes, making choices based on the breeder’s personal experience. Low

Method used

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  • Recombinant nucleic fragment RecCR010005 and detection primer and application thereof
  • Recombinant nucleic fragment RecCR010005 and detection primer and application thereof
  • Recombinant nucleic fragment RecCR010005 and detection primer and application thereof

Examples

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Effect test

Embodiment 1

[0068] Example 1 Breeding Recombinant Plants Introduced into Blast-resistant Genomic Fragments

[0069] The materials used in this example are rice 'Shen 95B' and rice 'Hua 3418B'.

[0070] The rice 'Hua 3418B' has good resistance to rice blast, and it is speculated that the region where the Pi1 and Pik cluster alleles of chromosome 11 play a key role in the rice blast resistance of this material.

[0071]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 11 from 27,155,000 to 28,495,000. 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 385 pairs of primers were designed. Through t...

Embodiment 2

[0081] Example 2 Determination of Homologous Recombination Fragments After Importing Rice Blast Resistance Genomic Fragments

[0082] In order to determine the size of the imported rice blast resistance genome fragment, the homozygous individual plants of the 'Shen 95B' 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 CR010005 was named RecCR010005.

[0083] It was preliminarily determined by the detection results of the rice genome-wide breeding chip RICE60K that RecCR010005 was located upstream of the SNP marker F1127618453CT.

[0084] At the same time, three samples of ‘Shen 95B’, ‘Hua 3418B’ and CR010005 were subjected to whole-genome sequencing using Miseq sequencing technology. The TruSeq Nano DNA LTKit (illumina) kit was used for library establishment, the Library QuantificationKit–Universal (KAPA Biosystems) kit was...

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Abstract

The invention relates to the molecular biology and specifically discloses a rice blast-resistant recombinant nucleic fragment RecCR010005 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. Shen 95B is used as the improved acceptor material and is a maintainer line matched with Shen 95A which is a widely used sterile line. Through the method, under the condition that the original advantages of the Shen 95B are reserved, the rice blast-resistant genome fragment is introduced, and the rice blast resistance of the Shen 95B can be greatly improved. The 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): C12N15/29C12N15/11C12Q1/68A01H1/02A01H1/04
CPCA01H1/02A01H1/04C12Q1/6895C07K14/415
Inventor 周发松喻辉辉张学堂邱树青孔会利雷昉姚玥李旭潘丽李菁韦懿陈光何予卿陈美容张启发
Owner CHINA NAT SEED GRP
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