Recombinant nucleic acid fragment RecCR010375 as well as 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 long time consumption and low efficiency, 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

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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 acid fragment RecCR010375 as well as detection primer and application thereof
  • Recombinant nucleic acid fragment RecCR010375 as well as detection primer and application thereof
  • Recombinant nucleic acid fragment RecCR010375 as well as detection primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Breeding 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. Thr...

Embodiment 2

[0085] Example 2 Determination of Homologous Recombination Fragments After Importing 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 CR010375 was named RecCR010375.

[0087] It was preliminarily determined by the rice genome-wide breeding chip RICE60K detection results that RecCR010375 was located downstream of the SNP marker F1127207217GA.

[0088] At the same time, three samples of 'Zhongzhonghui 629', 'Hua 130B' and CR010375 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 (KAPA Bio...

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 CR010375 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 molecular biology and specifically discloses a rice blast resistant recombinant nucleic acid fragment RecCR010375 as well as a detection primer and an application thereof. The invention further provides a method for breeding rice plants with rice blast resistant genome recombinant nucleic acid fragments based on a whole genome selective breeding technology. Target genome segments are imported into an acceptor material, and background restoration is realized. The improved acceptor material is 'Zhongzhonghui 629' and is a high-yield high-quality and optimal broad-spectrum restoration system. By utilizing the method, the rice blast resistance can be greatly improved under the condition that the original advantages of 'Zhongzhonghui 629' are remained. Furthermore, hybrid rice blast resistance is greatly improved through matching. The genome recombinant nucleic acid fragments provided by the invention are tightly related to the rice blast resistance and can serve as resistance resources to be applied to cultivation of other species.

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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IPC IPC(8): C12Q1/68C12N15/11A01H1/02A01H1/04
CPCA01H1/02A01H1/04C12Q1/6895C12Q2600/13
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杨毅张启发
Owner CHINA NAT SEED GRP
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