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Specific molecular markers for 1R-7R chromosomes of rye based on rye EST (expressed sequence tag) sequence and application of specific molecular markers

A molecular marker and chromosome technology, applied in the fields of molecular biology and genetics and breeding, to achieve the effect of low experimental technical requirements, strong conservation, and simple operation steps

Inactive Publication Date: 2011-09-14
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

[0004] 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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  • Specific molecular markers for 1R-7R chromosomes of rye based on rye EST (expressed sequence tag) sequence and application of specific molecular markers
  • Specific molecular markers for 1R-7R chromosomes of rye based on rye EST (expressed sequence tag) sequence and application of specific molecular markers
  • Specific molecular markers for 1R-7R chromosomes of rye based on rye EST (expressed sequence tag) sequence and application of specific molecular markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Development of rye 5R chromosome-specific molecular markers

[0035] 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 additional lines; among them, the "Chinese Spring-Imperial Rye" disomic additional 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:

[0036] A. Preparation of labeled primers: synthesized by Shanghai United Gene Co., Ltd. according to the sequence of 8-CGG16 labeled primers;

[0037] B. Dilution of labeled primers: Dissolve the labeled primers synthesized in the previous step to 100mmol L with 1×TE buffer -1 Store the mother liquor in...

Embodiment 2

[0043] Example 2: Development of rye chromosome-specific molecular markers for 1R and 6R

[0044] 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:

[0045] A. Preparation of labeled primers: synthesized by Shanghai United Gene Co., Ltd. according to the sequence of 1-CGG143 labeled primers;

[0046] B. Dilution of labeled primers: Dissolve the labeled primers synthesized in the previous step to 100mmol L with 1×TE buffer -1 Store the mother liquor in a refrigerator at -20°C, draw the mother liquor and dilute it with ultrapure water to 10mmol 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 EDT...

Embodiment 3

[0052] Example 3: Application of rye 1R and 6R chromosome-specific molecular markers

[0053] 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:

[0054] A. Preparation of labeled primers: synthesized by Shanghai United Gene Co., Ltd. according to the sequence of 1-CGG143 labeled primers;

[0055] B. Dilution of labeled primers: Diss...

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Abstract

The invention discloses 14 specific molecular markers for the 1R-7R chromosomes of rye based on a rye EST (expressed sequence tag) sequence, and the molecular markers are forward primers and reverse primers which are designed according to the EST sequence of rye chromosomes; and the molecular markers are used for identifying the specific markers of the 1R-7R chromosomes of rye. The identification of the 1R-7R chromosomes of rye by applying the molecular markers comprises the basic steps of: A. selecting and preparing the marker primers; B. diluting the marker primers; and C. marking the specificity of the rye chromosomes: C-1, extracting template DNA; C-2, carrying out PCR (polymerase chain reaction) amplification; C-3, detecting an amplified product; and C-4, comparing results. In the invention, a set of the specific molecular markers for the rye chromosomes are designed and developed on the basis of the EST sequence of the rye chromosomes, and the whole 1R-7R rye chromosomes in a genetic background of wheat can be rapidly and accurately identified by using a common PCR technology by means of the molecular markers, and thus the invention provides a simple and effective molecular identification method for the transfer of beneficial genes on the rye chromosomes to wheat cultivation.

Description

technical field [0001] 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 [0002] A relative of wheat, rye ( Secale cereale L.) has many excellent traits, such as well-developed root system, strong tillering, drought resistance, barren resistance, cold resistance, saline-alkaline and acidic soil resistance, especially resistance to various diseases, such as wheat powdery mildew ( Blumeria graminis f. sp. tritici ), stripe rust ( Puccinia graminis f. sp. tritici ), leaf rust ( P . triticina Eriks.) 、 Stem rust ( P . graminis Pers. f. sp. tritici ), Tilletia Controversa Kuhn (TCK) and barley yellow dwarf virus (Barley yellow dwarf virus, BYDV), etc. Therefore, rye is an important gene source for genetic improvement of wheat,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
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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