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Chromosome specific marker of elytrigia elongata in wheat background and use thereof

A technology of E. elongatum and specific markers, applied in the field of crop genetics and breeding, can solve the problems of small quantity, chromosome specificity and unsatisfactory stability, etc., and achieve the effect of simple operation, excellent marker resources, and superior stability

Inactive Publication Date: 2014-10-22
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, some chromosome-specific markers have been obtained through molecular marker development techniques such as RAPD, AFLP, and SSR. Chromosomal specificity and stability are not yet ideal. More molecular markers covering related chromosomes should be developed, and the linkage relationship between these molecular markers and disease resistance and stress resistance genes should be further studied, and these markers should be used in wheat stress resistance and disease resistance breeding. Assisted selection is very important and urgently needed

Method used

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  • Chromosome specific marker of elytrigia elongata in wheat background and use thereof
  • Chromosome specific marker of elytrigia elongata in wheat background and use thereof
  • Chromosome specific marker of elytrigia elongata in wheat background and use thereof

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Effect test

Embodiment 1

[0050] The design of embodiment 1 primer

[0051] Such as figure 1 Shown is the principle of primer design based on TRAP technology, 8 fixed primers were designed according to the EST sequence of Echinopsis elongatum, and the number of bases was between 18-19. figure 2 Shown is a total of 14 random primers designed according to the SRAP primers published in the literature (G.Li·C.F.Quiros, 2001), and the number of bases is between 17-19. Such as image 3 Shown are a total of 56 pairs of primers randomly combined by fixed primers and random primers. 56 primer combinations were combined with 18 basic primers for PCR amplification, and 21 primer combinations with specific amplification of E. elongatum were selected. The above primers are kept in the Laboratory of Molecular Cytogenetics, School of Biotechnology, Yangzhou University.

Embodiment 2

[0052] The extraction of embodiment 2 genome DNA

[0053] Genomic DNA was micro-extracted by the SDS phenol-chloroform method. The steps are as follows:

[0054] (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;

[0055] (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;

[0056]

[0057] (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;

[0058] (4) 12000rpm, centrifuge for 10min, draw the supernatant into a new centrifuge tube;

[0059] (5) Add about 750 μl of chloroform, turn it upside down, mix well, and extract for 5 minutes;

[0060] (6) 12000rpm, centrifuge for 10min, draw the supernatant in...

Embodiment 3

[0065] Example 3 PCR Amplification of Specific Fragment of Echinopsis elongatum

[0066] in accordance with image 3 PCR amplification was performed after the combination of primers. Such as Figure 4 Shown is a 25μl PCR reaction system, which is the most basic reaction system for PCR. Figure 5 Shown is a 35-cycle reaction program according to the TRAP technique in the literature (Hu et al., 2003). After the PCR reaction, the PCR product was detected by 6% polyacrylamide gel electrophoresis, such as Figure 6 Shown are the specific fragments of E. elongatum 1E-7E and the whole genome chromosome amplified by each primer combination, among which 1-7 are Chinese spring-Elongatum elongatum disomic addition lines, 8 is Chinese spring, and 9 is Elongatum elongatum Wheatgrass, A-G represent the specific markers of 1E-7E respectively, and H represents the specific markers of the whole genome. It can be seen from the figure that each chromosome has its own unique fragments, which ...

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Abstract

The invention relates to a chromosome specific marker of elytrigia elongata in wheat background and a use thereof. Primers are designed according plant EST sequence information, a wheat-elytrigia elongate addition line is amplified by a PCR technology to form a specific DNA fragment of elytrigia elongate, the specific DNA fragment is sequenced, novel primers are designed according to a sequence nonhomologous with wheat, and the elytrigia elongata chromosome specific molecule marker is obtained. The chromosome specific marker can be used for translocation line identification in chromosome fragment transfer from elytrigia elongata to wheat, can be used for gibberellic disease-resistant gene linkage analysis, can be used as a specific molecule marker for identification of elytrigia elongata chromosome and can be used for wheat gibberellic disease resistance and stress resistance breeding. The chromosome specific marker solves the problem that through a development technology of molecule markers such as RAPD, AFLP and SSR, specific markers of a part of chromosomes are obtained, the developed elytrigia elongata chromosome specific markers are less and cannot satisfy actual requirements on wheat resistance breeding and chromosome singularity and stability are non-ideal.

Description

technical field [0001] The invention belongs to the field of crop genetics and breeding, and in particular relates to a chromosome-specific marker of Echinopsis elongatum in the background of wheat and an application thereof. Background technique [0002] (1) Wheat breeding targets and their wild relatives [0003] Wheat is the second food crop in the world after corn. In the past half century, the world's total wheat production has more than tripled, and excellent wheat varieties have played a decisive role in increasing wheat production. [0004] Many wild relatives of wheat grow in nature, and these wild relatives contain a large number of good genes, which is a huge genetic gene pool with potential utilization value. Improvement will be of great value, not only to increase its yield and improve its quality, but also to transfer abundant disease-resistant and stress-resistant genes into wheat to improve the disease and stress resistance of wheat. At present, the genera...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 陈建民秦树文高勇陈士强张璐璐刘广晓
Owner YANGZHOU UNIV
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