Recombinant nucleic acid fragment RecCR012069 of rice genome and detection method of recombinant nucleic acid fragment RecCR012069

A technology for recombinant nucleic acid and genome, applied in the field of recombinant nucleic acid fragments and its detection, can solve the problems of low efficiency and long time consumption, achieve excellent cold resistance, and improve the effect of rice blast resistance

Active Publication Date: 2018-05-04
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 e

Method used

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  • Recombinant nucleic acid fragment RecCR012069 of rice genome and detection method of recombinant nucleic acid fragment RecCR012069
  • Recombinant nucleic acid fragment RecCR012069 of rice genome and detection method of recombinant nucleic acid fragment RecCR012069
  • Recombinant nucleic acid fragment RecCR012069 of rice genome and detection method of recombinant nucleic acid fragment RecCR012069

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Breeding of Recombinant Plants Introduced with Blast Resistance Genome Fragment

[0043] The materials used in this example are rice 'Kongyu 131' and rice 'Gumei 4'.

[0044] The rice 'Gumei 4' has good resistance to rice blast, and it is speculated that the Pi2 interval of chromosome 6 may play a key role in the rice blast resistance of this material.

[0045] 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 designed. The polymorphisms of the above prime...

Embodiment 2

[0055] Example 2 Determination of Homologous Recombination Fragments After Introducing Blast Resistance Genome Fragments

[0056] In order to determine the size of the imported rice blast resistance genome fragment, the homozygous single plant of the 'Kongyu 131' imported fragment 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 CR012069 was named RecCR012069.

[0057] It was preliminarily determined by the rice genome-wide breeding chip RICE60K detection results that the upstream homologous recombination fragment of RecCR012069 was located between markers F0610288021CT and R0610355182TC, and the downstream homologous recombination fragment was located between markers R0610435056GA and R0610486650CT.

[0058] At the same time, three samples of 'Kongyu 131', 'Gumei 4' and CR012069 were sequenced using Miseq sequencing technology. The TruSeq...

Embodiment 3

[0071] Example 3Resistance Identification of 'Kongyu 131' After Introducing Blast Resistance Genome Fragment

[0072] In order to identify the resistance effect, the new line CR012069 selected by the application, the recurrent parent 'Kongyu 131', 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:

[0073] The 14-7322-1 rice blast strain isolated from Heilongjiang in 2014 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 mycelium pieces with a diameter of 5 mm and trans...

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Abstract

The invention provides recombinant nucleic acid fragments of a rice genome and a detection method of the recombinant nucleic acid fragments and further provides a breeding method for rice plants containing the recombinant nucleic acid fragments. Foreground selection and background selection are performed on the recombinant plants by molecular markers, and the rice plants containing the recombinantnucleic acid fragments are obtained.

Description

technical field [0001] This application relates to genome-wide selective breeding technology. Specifically, the application relates to the selection and breeding of rice plants with recombinant nucleic acid fragments with rice blast resistance function by using genome-wide selective breeding technology, as well as the recombinant nucleic acid fragments obtained thereby and detection methods thereof. 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 m...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6895A01H1/02A01H1/04
CPCA01H1/02A01H1/04C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 周发松喻辉辉陆青刘刚陈伟康韦懿石义涛宋丁丁张龙雨雷昉姚玥李旭潘丽李菁陈光
Owner CHINA NAT SEED GRP
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