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Seeding method of target genome DNA fragment-containing plants

A genome, whole genome technology, applied in the fields of genetics, genomics and molecular breeding, can solve the problems of offspring phenotype separation, lack of stability, judgment, etc., to speed up the breeding process and improve the breeding efficiency.

Active Publication Date: 2016-05-11
CHINA NAT SEED GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the genetic background of the selected plants cannot be judged at the genome-wide level, there is a risk of loss of some excellent traits of the recipient parents
(3) Lack of stability
Breeding plants may cause segregation of offspring phenotypes due to genotype impurity

Method used

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  • Seeding method of target genome DNA fragment-containing plants
  • Seeding method of target genome DNA fragment-containing plants
  • Seeding method of target genome DNA fragment-containing plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Screening of recombined single plant of rice 'Kongyu 131' introduced with DNA fragment of blast resistance gene

[0060] The materials used in this example are rice 'Kongyu 131' and 'K22'. 'K22' has been proven to have good blast resistance, and it is speculated that the gene cluster region where Pi2, Pi9 and Pigm of chromosome 6 are located may play a key role in the rice blast resistance of this material.

[0061] The inventor introduced the genomic DNA fragment where the above-mentioned gene clusters in 'K22' are located into 'Kongyu 131', the specific process is as follows:

[0062] Screening of foreground selection markers: positive selection markers are to screen polymorphic molecular markers within the range of 50kb (in rice, its genetic distance is 0.2cM) on the upper and lower reaches of the target genome DNA (rice blast resistance gene DNA) fragment, and obtain the same Tightly linked markers Pi2-4 for the target genomic DNA fragment. The negative ...

Embodiment 2

[0069] Example 2 Determination of homologous recombination fragments after 'Kongyu 131' was introduced into rice blast resistance gene

[0070] In order to determine the size of the imported rice blast resistance gene fragment and to protect the imported fragment, the inventors carried out homology of the target genome DNA fragment on both sides of the homozygous single plant of 'Kongyu 131' into which the rice blast resistance gene DNA fragment was introduced. Sequencing of recombinant fragments. First of all, it was preliminarily confirmed by the SNP chip detection results that the upstream homologous recombination fragment of the A08-1 rice blast resistance gene recombination DNA fragment segment was located in the approximately 68.2kb region between the two SNP markers R0610132417TC and R0610200630GA (10132417bp~10200630bp of chromosome 6) , the downstream homologous recombination fragment is located in the approximately 68.9kb region between the two SNP markers F061071165...

Embodiment 3

[0078] Example 3 Resistance Identification of 'Kongyu 131' After Introducing the Recombinant DNA Fragment of the Rice Blast Resistance Gene

[0079] In order to identify the resistance effect of the target individual plants, A08-1, 'Kongyu 131', Gumei No. 4 and Lijiang Xintuan Heigu were planted indoors, and they were cultivated to the 3-4 leaf stage for identification using the following method :

[0080] Ten strains of Magnaporthe oryzae isolated from diseased leaves of rice blast collected in the disease nursery of Heilongjiang Academy of Agricultural Sciences and Qixing Farm in Heilongjiang Province in 2013 were used as inoculation strains. Numbers 13-7101, 13-7102, 13-7103, 13-7104, 13-7105, 13-7201, 13-7202, 13-7203, 13-7204 and 13-7205. The strains are stored at -20°C by the paper disc method. Before use, take out the disc and activate it on PDA medium (peeled potato 200g, glucose 20g, agar powder 15g). After 5 days, take a fresh mycelium block with a size of 1 cm squa...

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Abstract

The invention provides a method for rapidly and accurately seeding target genome DNA fragment-containing plants on the basis of a whole-genome seeding technology. The method comprises the following steps: 1, hybridizing, backcrossing and selfing a donor plant parent containing the fragment and a recurrent parent which is a receptor plant parent free from the target genome DNA fragment; 2, using a foreground selection marker in the breeding process to carry out foreground selection; 3, using a high density molecular marker detection technology in the breeding process to carry out whole genome background selection; and 4, using above steps until the target plants with homologous recombination of two sides of the target genome DNA fragment, target genome DNA fragment homozygosis and complete background return. The invention also provides a seeding method of blast-resistant gene recombinant DNA fragment-containing plants. The stable plants having background being completely consistent with that of the receptor parent and only containing the donor parent target genome DNA fragment can be rapidly obtained through accurate foreground selection and high density molecular marker whole genome background selection.

Description

technical field [0001] The invention relates to the fields of genetics, genomics and molecular breeding, in particular to a whole-genome selective breeding technique and a method for quickly and accurately selecting plants containing target genome DNA fragments using the technique. 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, and its biggest disadvantage is that it takes a long time. In recent years, breeders have begun to use molecular marker-assisted selection to improve individual target traits. Although the application of this method can significantly shorten the breeding period, it still has the following disadvantages. (1) Introduction of non-target genomic DNA fragments. The application of existing molecular marker-assisted selection methods is mostly limited to the selection of a single locus, only ensurin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A01H1/02A01H1/04
Inventor 周发松喻辉辉刘刚杨燕宇韦懿陈光姚玥李菁宋丁丁李旭潘丽张学堂陆青邱树青何予卿李荣田张启发
Owner CHINA NAT SEED GRP
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