Specific mark of thinopyrum elongatum chromosome in wheat background and application thereof

A technology of E. elongatum and specific markers, applied in the field of crop genetics and breeding, to achieve superior stability, excellent marker resources, and good repeatability

Inactive Publication Date: 2012-02-15
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Specific mark of thinopyrum elongatum chromosome in wheat background and application thereof
  • Specific mark of thinopyrum elongatum chromosome in wheat background and application thereof
  • Specific mark of thinopyrum elongatum chromosome in wheat background and application thereof

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Effect test

Embodiment 1

[0068] Embodiment 1 plant material and PCR primer sequence

[0069] (1) Plant material

[0070] The materials used in the present invention include 7 parts of Chinese spring wheat-Thinopyrum elongatum disomic additional lines (Triticum aestivum cv. Chinese Spring-Thinopyrum elongatum disomic additional lines, DA1E, DA2E, DA3E, DA4E, DA5E, DA6E, DA7E,), 2 parts Chinese spring wheat-Thinopyrum elongatum 7E long arm and 7E short arm addition lines (DA7ES, DA7EL), 19 Chinese spring wheat-Thinopyrum elongatum disomic substitution lines (Triticum aestivum cv.Chinese Spring-Thinopyrum elongatum disomic substitutional lines, Such as DS1E / 1A, DS2E / 2A, DS3E / 3A to DS3E / 19A, 1 part of wheat Chinese spring (Triticum aestivum) and 1 part of diploid Elongated wheatgrass. The above materials are donated and preserved by Dr.Fedax of the Canadian Department of Agriculture In the Laboratory of Molecular Cytogenetics, School of Biotechnology, Yangzhou University.

[0071] (2) PCR amplification ...

Embodiment 2

[0077] The extraction of embodiment 2 genome DNA

[0078] DNA was micro-extracted by the SDS phenol-chloroform method. The steps are as follows:

[0079] (1) Take the young leaves (about 0.1g), cut them into pieces and put them into a 2ml centrifuge tube, cool them in liquid nitrogen, and grind them to powder with a grinding rod;

[0080] (2) Place the centrifuge tube at room temperature to cool slightly, add 700 μl of buffer A, mix gently, then bathe in water at 65°C for 20 minutes, and mix by inverting up and down every 5 minutes;

[0081] Buffer A:

[0082]

[0083] Dilute the volume to 1L with ddH2O and use it after sterilization.

[0084] (3) Take it out and cool it down to room temperature, add 350 μl of phenol and chloroform each, turn it upside down, mix well, and extract for 5 minutes;

[0085] (4) 12000rpm, centrifuge for 10min, draw the supernatant into a new centrifuge tube;

[0086] (5) Add about 750 μl of chloroform, turn it upside down, mix well, and ext...

Embodiment 3

[0092] Example 3 PCR Amplification of Specific Fragment of Echinopsis elongatum

[0093] According to the combination of basic primers in Table 1, PCR amplification was carried out. The specific reaction system and reaction program of PCR are shown in Table 3 and Table 4. After the PCR reaction, the PCR products were detected by 6% polyacrylamide gel electrophoresis ( figure 1 ), the specific fragment sizes and specific chromosomes of E. elongatum amplified by each primer combination are shown in Table 2. Since there is a complete set of Chinese spring-Thinopyrum elongata disomic addition line materials, the amplified specific fragments can be further mapped to different E chromosomes.

[0094] Table 3 PCR reaction system

[0095]

[0096] Table 4PCR reaction program

[0097]

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Abstract

The invention belongs to the field of crop genetic breeding. Particularly, a primer is designed according to repeated sequence information of a plant; an additional wheat-thinopyrum elongatum system is amplified by utilizing a PCR (polymerase chain reaction) technology to obtain and sequence a specific DNA (deoxyribonucleic acid) segment of thinopyrum elongatum; the primer is designed again according to the sequence without homology to that of the wheat so as to obtain the specific molecule mark of thinopyrum elongatum chromosome, wherein the molecule mark is one of eight pairs of primers provided by the invention. The marks can be used for translocation line identification in the process of transferring chromosome segments from thinopyrum elongatum to wheat, can be used for chain analysis of a gibberellic disease resistant gene, and can be used as the specific molecule marks to identify thinopyrum elongatum chromain for gibberellic disease resistance and stress resistant breeding of wheat.

Description

technical field [0001] The invention belongs to the field of crop genetics and breeding. The primers are designed according to the repeated sequence information of plants, and the specific fragments of Thinpyrum elongatum are obtained by amplifying and sequencing them from the additional line of wheat-Thinopyrum elongatum by using PCR technology, and the primers are redesigned according to the sequencing results. Chromosome-specific molecular markers were obtained from E. elongatum. These markers can be used for the identification of translocation lines in the process of transferring chromosome segments from E. elongatum to wheat, for the linkage analysis of wheat scab resistance and stress resistance genes, and for the identification of wheat scab resistance and stress resistance breeding. Molecular marker-assisted selection. Background technique [0002] (1) Wheat breeding targets and their wild relatives [0003] Wheat is the second food crop in the world after corn. I...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 陈建民朱雪高勇陈士强张超葛江燕
Owner YANGZHOU UNIV
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