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Inter-simple sequence repeat ISSR-SCAR marker specific to E-group chromosomes of agropyron elongatum

A technology of Echinopsis elongatum and chromosomes, which is applied to the determination/testing of microorganisms, biochemical equipment and methods, etc., and can solve the problems of poor repeatability and stability

Inactive Publication Date: 2013-01-23
HARBIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its repeatability and stability are poor. If it is converted into a SCAR marker, its specificity and stability will be greatly improved, and it can be more conveniently and quickly applied to the detection of heterologous chromosomes or chromatin

Method used

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  • Inter-simple sequence repeat ISSR-SCAR marker specific to E-group chromosomes of agropyron elongatum
  • Inter-simple sequence repeat ISSR-SCAR marker specific to E-group chromosomes of agropyron elongatum
  • Inter-simple sequence repeat ISSR-SCAR marker specific to E-group chromosomes of agropyron elongatum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Obtaining of Chromosome-Specific ISSR-SCAR Marker UBC841 of Group E of E. elongatum

[0033] 1. Screening and Cloning of Chromosome-Specific ISSR-SCAR Marker UBC841 in Group E of E. elongatum

[0034] Using 33 ISSR primers, the materials such as common wheat "China Spring", "China Spring"-gametocidal chromosome 2C disomy addition line, diploid Henoptera elongatum and tetraploid Henna elongatum were amplified and screened, Isolation and cloning of specific amplified fragments of the E genome.

[0035] PCR amplification system:

[0036] Template DNA 50ng

[0037] 10x buffer 2.5μl

[0038] 1.5mM MgCl 2 2.5μl

[0039] dNTP Mix (2.5mM) 1.5μl

[0040] ISSR Primer (10μM) 1μl

[0041] r Taq polymerase (5U / μl) 0.25μl

[0042] Distilled sterile water up to 20μl

[0043] PCR reaction program: pre-denaturation at 94°C for 4 minutes, 42 cycles including denaturation at 94°C for 1 minute, annealing at 52°C for 1 minute, extension at 72°C for 2 min...

Embodiment 2

[0067] Example 2, Application of Chromosome-Specific ISSR-SCAR Markers in Group E of E. elongatum

[0068] use SCAR 577 Upstream primers and SCAR 577 F 1and inbred F 2 Generation was identified. Common wheat "China Spring", "China Spring"-gametocidal chromosome 2C disomy addition line, diploid E. elongatum, and tetraploid E. elongatum were used as controls.

[0069] The result is as image 3 As shown, 1-9. is the F obtained by crossing the "China Spring"-Thinopyrum elongatum disomic addition line and "China Spring"-gametocidal chromosome 2C disomic addition line 1 generation; 10-19. is F 1 F 2 generation; 20. is the common wheat "China Spring"; 21. is the "China Spring"-gametocidal chromosome 2C disomy addition line; 22. is the diploid E. elongatum; 23. is the tetraploid E. elongatum ; M. is DL2000marker; it can be seen from the figure that SCAR 577 marked in F 1 All generations showed positive results (a 577bp fragment was obtained). In order to carry out preliminar...

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Abstract

The invention discloses an inter-simple sequence repeat ISSR-SCAR marker specific to E-group chromosomes of agropyron elongatum. The invention provides a kit for detecting the E-group chromosomes of agropyron elongatum. The kit comprises the following primer pair: one primer in the primer pair is nucleotide shown in a sequence 3 of a sequence table, and the other primer in the primer pair is nucleotide shown in a sequence 4 of the sequence table. The E-group chromosomes of thinopyrum elongatum are the basic chromosome group forming the polyploid species of elytrigia and carry the genes beneficial to the genetic breeding of wheat, the detection of exogenous E chromatins in wheat is the key point for identifying recombination lines and introgression lines of the wheat, namely the agropyron elongatum, the work of screening and identifying a large number of filial generations is quite complicated, and tests prove that the ISSR-SCAR marker developed by the invention can lay a foundation for quickly and accurately identifying the E-group chromatins or chromosomes of exogenous agropyron elongatum under the genetic background of wheat.

Description

technical field [0001] The invention relates to a specific ISSR-SCAR marker for chromosome E of E. elongatum, in particular to a kit for detecting chromosome E of wheat and its application. Background technique [0002] Enopyrum is a close relative of wheat. It has many excellent traits that common wheat lacks, such as strong tillering ability, well-developed root system, cold resistance, drought resistance, barren resistance, strong salt and alkali resistance, multiple flowers, large spikes, Fruity, high grain protein content and excellent baking quality, strong resistance to various diseases, high resistance to wheat stripe rust, leaf rust, stem rust, powdery mildew, yellow dwarf disease, stripe mosaic disease and tickle smut To immunity, it is one of the important parents of wheat genetics and breeding. Agropyron elongatum (Agropyron elongatum) is an important close relative of wheat. It has excellent properties such as high protein content, resistance to various fungal ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 郭长虹徐国辉束永俊司亮王晓萍
Owner HARBIN NORMAL UNIVERSITY
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