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A method for creating and identifying a chromosomal translocation line between a and b genomes of peanut

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: 2021-02-05
HENAN ACAD OF AGRI SCI
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  • Summary
  • 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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  • A method for creating and identifying a chromosomal translocation line between a and b genomes of peanut
  • A method for creating and identifying a chromosomal translocation line between a and b genomes of peanut

Examples

Experimental program
Comparison scheme
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 identifying a chromosomal translocation line between the genomes of peanut A and B. The method for creating is to use the cultivar Silihong as the material, and use Co 60 ‑γ-ray irradiated peanut plants to obtain peanut plants with chromosomal translocation between A and B genomes. The present invention adopts florescence Co 60 ‑γ-irradiation induced a large number of translocations between genomes A and B, realizing chromatin exchange between genomes A and B, providing new resource materials for peanut breeding and molecular cytogenetic research, and also for distant hybridization Artificial creation of exogenous chromosome (segment) introgression lines in progeny materials provides a way. The present invention uses genome fluorescence in situ hybridization technology to identify chromosome translocation lines between peanut A and B genomes, creates and identifies a batch of peanut cultivars A and B genome chromosome translocation lines new materials for the first time, and provides a basis for identifying peanut chromosome translocations effective means.

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 Patents(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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