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Secale cereale EST (expressed sequence tag) sequence based specific molecular markers of secale cereale 1R and 6R chromosomes and application thereof

A molecular marker and specific marker technology, applied in the fields of molecular biology and genetic breeding, to achieve the effect of rich marker sources, strong binding ability and strong conservation

Inactive Publication Date: 2013-02-13
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, so far, no one has reported a complete set of rye 1R-7R chromosome-specific molecular markers derived from the rye expressed sequence tag (EST) sequence to detect and identify all 1R-7R in the wheat background. rye chromosome

Method used

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  • Secale cereale EST (expressed sequence tag) sequence based specific molecular markers of secale cereale 1R and 6R chromosomes and application thereof
  • Secale cereale EST (expressed sequence tag) sequence based specific molecular markers of secale cereale 1R and 6R chromosomes and application thereof
  • Secale cereale EST (expressed sequence tag) sequence based specific molecular markers of secale cereale 1R and 6R chromosomes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Development of rye 5R chromosome-specific molecular markers

[0037] Using 8-CGG16 molecular marker to identify the 5R chromosome of rye, the test material is the disomic addition line of the 5R chromosome of "Chinese spring-imperial rye", and the control materials are imperial rye, common wheat Chinese spring and The remaining 6 "Chinese Spring-Imperial Rye" disomic addition lines; among them, the "Chinese Spring-Imperial Rye" disomic addition lines (1R-7R) are known public materials (references: Driscoll C J, Sears E R , 1971.Individual additions of the chromosomes of "Imperial" rye to wheat.Agron Abstr, 1971: 6); the operation steps are as follows:

[0038] A. Preparation of labeled primers: synthesized by Shanghai United Gene Co., Ltd. according to the sequence of 8-CGG16 labeled primers;

[0039] B. Dilution of labeled primers: Dissolve the labeled primers synthesized in the previous step to 100 μmol L with 1×TE buffer -1 Store the mother liquor in a -...

Embodiment 2

[0045] Example 2: Development of rye chromosome-specific molecular markers for 1R and 6R

[0046] Using 1-CGG143 marker to identify the 1R and 6R chromosomes of rye, the test material is the disomic addition line of 1R and 6R chromosomes of "Chinese Spring-Imperial Rye", and the control materials are Imperial Rye and Common Wheat China Spring and the other 5 "China Spring-Imperial Rye" disomic addition lines; the operation steps are as follows:

[0047] A. Preparation of labeled primers: synthesized by Shanghai United Gene Co., Ltd. according to the sequence of 1-CGG143 labeled primers;

[0048] B. Dilution of labeled primers: Dissolve the labeled primers synthesized in the previous step to 100 μmol L with 1×TE buffer -1 Store the mother liquor in a -20°C refrigerator, draw the mother liquor and dilute it with ultrapure water to 10 μmol L before use -1 The working solution is ready for use; the formula of 1×TE buffer solution is, 10mmol L -1 Tris-HCl with 1mmol L -1 EDTA, ...

Embodiment 3

[0054] Example 3: Application of rye 1R and 6R chromosome-specific molecular markers

[0055] Using 1-CGG143 marker to identify rye 1R and 6R chromosomes in common wheat "Xiaoyan 6" and "German white" rye hybrid progeny, etc., the material to be tested is German white rye, WR64 (Xiaoyan No. 6 - German White 1BL / 1RS translocation line), WR81 (Xiaoyan No. 6 - German White 1BL / 1RS translocation line), WR91 (Xiaoyan No. 6 - German White 2R (2D) substitution line), WR41 ( Xiaoyan 6-German White 4B / 4R reciprocal translocation line); control materials are Chinese spring, Xiaoyan 6, Imperial rye, Lovelin 10 (1BL / 1RS translocation line of Petkus rye), Love Lin 13 (1BL / 1RS translocation line of Petkus rye), triticale 06CT456 (AABBRR), triticale 06CT461 (AABBRR); the operation steps are as follows:

[0056] A. Preparation of labeled primers: synthesized by Shanghai United Gene Co., Ltd. according to the sequence of 1-CGG143 labeled primers;

[0057] B. Dilution of labeled primers: Diss...

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Abstract

The invention discloses secale cereale EST (expressed sequence tag) sequence based specific molecular markers of secale cereale 1R and 6R chromosomes, which are forward primers and reverse primers designed according to ESTs on secale cereale chromosomes, and are the specific markers used for identifying the secale cereale 1R and 6R chromosomes. The basic steps used for identifying the secale cereale 1R and 6R chromosomes through the application of the molecular markers are as follows: A, selecting and preparing marked primers; B, diluting the marked primers; and C, the specific markers of the secale cereale 1R and 6R chromosomes: C-1, extracting DNA of templates, C-2, performing PCR (polymerase chain reaction) amplification, C-3, detecting amplified products, and C-4, performing results comparison. By virtue of the molecular markers provided by the invention, the ordinary PCR technology can be adopted to quickly and accurately identify the secale cereale 1R and 6R chromosomes in a wheat genetic background, so that a simple, convenient and effective molecular identification method is provided for transferring useful genes on the secale cereale chromosomes into a wheat cultivation background.

Description

[0001] This patent application is a divisional application of the following patent: the title of the invention is "Rye 1R-7R chromosome-specific molecular markers based on rye EST sequence and its application", the application date is 2011-04-14; the application number is 201110092989. X. technical field [0002] The invention relates to the fields of molecular biology and genetic breeding, in particular to molecular markers designed based on rye expressed sequence tag (EST) sequences, and the application of these molecular markers in tracking and detecting rye chromosomes. Background technique [0003] Rye (Secale cereale L.), a close relative of wheat, has many excellent characters, such as developed root system, strong tiller, drought resistance, barren resistance, cold resistance, salt-alkaline and acid soil resistance, especially resistance to various diseases, such as wheat flour (Blumeria graminis f.sp.tritici), stripe rust (Puccinia graminis f.sp.tritici), leaf rust ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 安调过尹冬冬许红星李立会
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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