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Recombinant nucleic fragment RecCR010374 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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  • 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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  • Recombinant nucleic fragment RecCR010374 and detection primer and application thereof
  • Recombinant nucleic fragment RecCR010374 and detection primer and application thereof
  • Recombinant nucleic fragment RecCR010374 and detection primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Breeding of Recombinant Plants Introduced into Blast-resistant Genomic Fragments

[0073] The materials used in this example are rice 'Zhongzhonghui 629' and rice 'Hua 130B'.

[0074] The rice 'Hua 130B' 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.

[0075] 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. ...

Embodiment 2

[0085] Example 2 Determination of Homologous Recombination Fragments After Introduction of Rice Blast Resistance Genomic Fragments

[0086] 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 CR010374 was named RecCR010374.

[0087] It was preliminarily determined by the rice genome-wide breeding chip RICE60K detection results that RecCR010374 was located downstream of the SNP marker F1127345466CT.

[0088] At the same time, three samples of 'Zhongzhonghui 629', 'Hua 130B' and CR010374 were subjected to whole-genome sequencing using Miseq sequencing technology. The TruSeq Nano DNA LTKit (illumina) kit was used for library establishment, the Library Quantification Kit–Universal (KA...

Embodiment 3

[0097] Example 3 Resistance Identification of 'Zhongzhonghui 629' After Introducing the Rice Blast Resistance Genome Fragment

[0098] In order to identify the resistance effect, the new line CR010374 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:

[0099] Select 21K02-1 and 21K14-1 isolated from rice blast samples from Enshi disease nursery in 2013, 6102-1 from Sichuan disease nursery, 8111-1 from Fujian disease nursery, and rice blast disease samples from Enshi disease nursery in 2011 Isolated LJ7-1-2, a total of 5 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...

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Abstract

The invention relates to the molecular biology and specifically discloses a rice blast-resistant recombinant nucleic fragment RecCR010374 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. Zhongzhonghui 629 is used as the improved acceptor material and is a strong broad-spectrum restoring line with a high yield and high quality. Through the method, the rice blast resistance of the Zhongzhonghui 629 can be greatly improved under the condition that the original advantages of the Zhongzhonghui 629 are reserved. 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
CPCA01H1/02A01H1/04C12Q1/6895C12Q2600/156
Inventor 喻辉辉周发松张学堂邱树青何宗顺雷昉姚玥潘丽李旭李菁韦懿陈光何予卿杨毅张启发
Owner CHINA NAT SEED GRP
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