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Recombined nucleic acid fragment reccr010161 and its detection primers and applications

A technology of fragment and recombination single plant, applied in the field of molecular biology, can solve the problems of long time and low efficiency.

Active Publication Date: 2021-12-10
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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  • Recombined nucleic acid fragment reccr010161 and its detection primers and applications
  • Recombined nucleic acid fragment reccr010161 and its detection primers and applications
  • Recombined nucleic acid fragment reccr010161 and its detection primers and applications

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 'Luxiang 618B' and rice 'Hua 3418B'.

[0074]Rice 'Hua 3418B' has good blast resistance, and it is speculated that the region where the Pil 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. Through the PC...

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 single plant of the imported fragment of 'Luxiang 618B' was 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 CR010161 was named RecCR010161.

[0087] It was preliminarily determined by the rice genome-wide breeding chip RICE60K detection results that RecCR010161 was located downstream of the SNP marker R1127353173AG.

[0088] At the same time, three samples of 'Luxiang 618B', 'Hua 3418B' and CR010161 were subjected to whole-genome sequencing using Miseq sequencing technology. The TruSeq Nano DNA LT Kit (illumina) kit was used for library establishment, the LibraryQuantification Kit–Universal (KAPA Biosystems) ...

Embodiment 3

[0097] Example 3 Resistance Identification of 'Luxiang 618B' After Introducing the Rice Blast Resistance Genome Fragment

[0098] In order to identify the resistance effect, the new line CR010161 selected by the application, the recurrent parent 'Luxiang 618B', 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:

[0099] M15Bb-1-1, M15Bb-1-2, M15Bb-2-1, M15Bb-3-1, M15Bb-4-1, M15Bb-5-1, M15Bb- 6-1, a total of 7 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 (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 molecular biology, and specifically discloses a rice blast-resistant recombinant nucleic acid fragment RecCR010161, detection primers and applications thereof. The present invention also provides a method for breeding rice plants containing blast-resistant genome recombined nucleic acid fragments based on the whole-genome selective breeding technique, in which the target genome fragments are introduced into recipient materials, and the background is restored at the same time. The improved acceptor material of the present invention is 'Luxiang 618B', which is a maintainer line for the fragrant CMS line 'Luxiang 618A' with medium and low amylose content. Using the above method, the rice blast resistance of 'Luxiang 618B' can be greatly improved while retaining the original advantages. Further, the blast resistance of the hybrid rice is greatly improved through mating. The genome recombinant nucleic acid fragment provided by the invention is closely related to rice blast resistance, and can be used as a resistance resource for the 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 Patents(China)
IPC IPC(8): C12N15/29C12N15/11C12Q1/6895C12Q1/686A01H1/02A01H1/04A01H5/00A01H6/46
CPCA01H1/02A01H1/04C12Q1/6895C07K14/415
Inventor 周发松喻辉辉张学堂邱树青张龙雨雷昉姚玥李旭潘丽李菁韦懿陈光何予卿陈国良张启发
Owner CHINA NAT SEED GRP
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