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Method for creating and authenticating chromosome translocation line between peanut genomes A and B

A technology of chromosomal translocation and identification method, which is applied in the field of creation and identification of chromosomal translocation lines between peanut A and B genomes, and can solve the problems of unseen peanut chromosomal translocation and lack of research on peanut genome chromosomal translocation.

Active Publication Date: 2018-01-19
HENAN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In peanut radiation mutagenesis, the use of 60 Co-γ rays irradiate peanut seeds, and their oleic acid content, linoleic acid content, oil content and other traits have changed significantly, but there have been no reports of inducing chromosomal translocations between peanut A and B genomes, and there is also a lack of utilization. 60 Study on Chromosomal Translocation of Peanut Genome Created by Co-γ Rays

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  • Method for creating and authenticating chromosome translocation line between peanut genomes A and B
  • Method for creating and authenticating chromosome translocation line between peanut genomes A and B

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Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1, the creation method of chromosomal translocation line between peanut A and B genomes

[0047] Experimental materials: peanut cultivar "Si Li Hong" (allotetraploid 2n=4x=40), preserved by the Economic Crops Research Institute of Henan Academy of Agricultural Sciences.

[0048] The experimental materials were planted in flower pots, using Co 60 - Gamma ray irradiation and identification by genome fluorescence in situ hybridization to obtain the chromosomal translocation line between the A and B genomes. The specific method is as follows:

[0049] (1) Plant the peanut cultivar "Si Lihong" in a flowerpot, and from the first day of flowering of the plant, pinch off the newly opened flowers from the base of the flower stalk every morning to control the fruiting of the flowers; Flowering period (the flowering amount is 50% of the total flowering amount), using Co 60 - Irradiation of whole peanut plants with gamma rays, at Co 60 - γ-ray (Henan Academy of Scien...

Embodiment 2

[0051] Embodiment 2, identification method of chromosomal translocation line between peanut A and B genomes

[0052] The identification method of the chromosomal translocation line between the peanut A and B genomes, specifically:

[0053] (1) Probe labeling: Genome donor ancestor species A. duranensis and A. duranensis and A. Whole genome DNA, and A. duranensis whole genome DNA was labeled with fluorescein by the nick translation method to obtain a fluorescein-labeled A. duranensis DNA probe; for A. The whole genome DNA was labeled with digoxin to obtain digoxin-labeled A. DNA probe; the specific method is:

[0054] Fluorescein-labeled A. duranensis DNA probe: First prepare the reaction solution in a 0.2mL eppendorf tube, add 800ng / μL A.duranensis DNA 3μL, 10×DNA Polymerase I Buffer 2.5μL, 1.3mMdNTP mix 2.5μL, 5U / μL DNaseI 0.5μL, 1mM Fluoresceint-12-dUTP 0.7μL, 5U / μL DNAPolymerase I 1μL, 14.8μL ddH 2 O, mix the reaction solution evenly, incubate at 16°C for 25 minutes,...

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Abstract

The invention discloses a method for creating and authenticating chromosome translocation line between peanut genomes A and B. The creating method comprises the steps of taking a peanut cultivation variety four-grain red as a material and irradiating the peanut plant by Co60-gamma rays at full-blossom period to obtain the peanut plant with the chromosome translocation between genomes A and B. Flowering phase Co60-gamma radiation induction is adopted, so that a large amount of chromosome translocation between the genomes A and B is generated, chromatin communication between the genomes A and Bis realized, a new resource material is provided for peanut breeding and molecular cytogenetic research, and an approach is provided for an artificially created exogenous chromosome (fragment) introgression line in a distant hybridization progeny material. The chromosome translocation line between peanut the peanut genomes A and B is authenticated by a genome fluorescent in-situ hybridization technology, a peanut cultivation variety genome A and B chromosome translocation line new material is created and authenticated for the first time, and an effective means is provided for authenticating peanut chromosome translocation.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology and relates to a method for creating and identifying a chromosome translocation line between A and B genomes of peanuts. Background technique [0002] Peanut (Arachis hypogaea L., 2n=4x=40, AABB) is an important oil crop in the world. During the long-term artificial selection and domestication process, the genetic diversity of peanut cultivars has decreased, and the genetic basis of breeding parents has become increasingly narrow, which has seriously affected the breeding efficiency. It is particularly important to create new germplasm and discover new genes. [0003] The peanut cultivars are allotetraploid, derived from the diploid wild species A. duranensis and A. After accidental hybridization, then doubled to form tetraploid wild peanut A. monticola, and finally evolved through artificial domestication. After long-term systematic research, it is generally believed that A. duranensi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895
Inventor 张新友杜培付留洋李丽娜韩锁义刘华秦利张忠信黄冰艳董文召汤丰收
Owner HENAN ACAD OF AGRI SCI
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