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Identification method for amblyopyrum muticum chromosome in wheat genome

A technology of goat grass and identification method, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., which can solve the problems of cumbersome operation steps, long and short length of blocked DNA, etc., and broaden the scope of application Effect

Inactive Publication Date: 2015-07-22
CROP RES INST SHANDONG ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

[0005] Genomic in situ hybridization (GISH) can be used to identify the chromosomes of foreign species in wheat background, but this method involves the extraction and fragmentation of genomic DNA of foreign species, the preparation of blocked DNA, the ratio of the two Conditional exploration and other steps, the operation steps are cumbersome, and the length of broken foreign species DNA and blocked DNA often appear too long or too short (the length is not suitable for GISH), and it is difficult to guarantee each time The length of fragmented DNA fragments is exactly the same. Therefore, it has obvious disadvantages compared with the fluorescence in situ hybridization (FISH) using polynucleotides as probes. The latter has the advantages of easy synthesis of probes and no need for sealing. DNA blocking, good reproducibility, simple operation, and clearly visible fluorescent signals, etc. However, this method has not been used for the identification of the chromosomes of A. amansifolia in wheat background.

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  • Identification method for amblyopyrum muticum chromosome in wheat genome
  • Identification method for amblyopyrum muticum chromosome in wheat genome
  • Identification method for amblyopyrum muticum chromosome in wheat genome

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Embodiment Construction

[0043] Hybrid F 1 ; Hybrid F 1 After doubling, the innovative germplasm of Chinese spring-goat grass (No. XX001) was obtained. The material numbered XX002-XX031 is the hybrid backcross of XX001 and CS (BC 2 f 5 ) offspring. Wheat China Spring (CS) was provided by Professor Yang Zujun, School of Life Science and Technology, University of Electronic Science and Technology of China. Goatgrass (TA2193) was provided by Dr. Raupp J, Center for Wheat Genomics and Genetic Resources, Kansas State University, USA. Probe Oligo-pSc119.2-1, probe Oligo-pTa535-1 and (GAA)8 were synthesized by Chengdu Ruixin Biological Company. The 3 probe sequences are as follows:

[0044] Probe Oligo-pSc119.2-1 sequence:

[0045] CCGTTTTGTGGACTATTACTCACCGCTTTGGGGTCCCATAGCTAT, as shown in SEQ ID NO.1;

[0046] Probe Oligo-pTa535-1 sequence:

[0047] AAAAACTTGACGCACGTCACGTACAAATTGGACAAACTCTTTCGGAGTATCAGGGTTTC, as shown in SEQ ID NO.2;

[0048] Probe (GAA) 8 sequence: GAAGAAGAAGAAGAAGAAGAAGAA, as show...

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Abstract

The invention relates to an identification method for an amblyopyrum muticum chromosome in a wheat genome, and belongs to the field of cell genetics. The identification method comprises the following steps: using Oligo-pTa535-1, Oligo-pSc119.2-1 and (GAA)8 as probes to perform fluorescence in-situ hybridization (FISH) on innovative idioplasmatic chromosome of wheat-amblyopyrum muticum, comparing with a bi-color FISH standard drawing of 42 chromosomes of the wheat, and at last obtaining hybridization signals and karyotype information of seven pairs of chromosomes of amblyopyrum muticum. According to the identification method disclosed by the invention, Oligo-pTa535-1, Oligo-pSc119.2-1 and (GAA)8 are used as the probes to perform fluorescence in-situ hybridization on chromosomes of wheat-amblyopyrum muticum filial generation; the results are respectively compared with the wheat-amblyopyrum muticum innovative idioplasmatic chromosome hybridization signals and karyotype information of the amblyopyrum muticum chromosomes, and at last the identification is completed. The invention provides the novel method for detecting the amblyopyrum muticum chromosome in the wheat genome.

Description

technical field [0001] The invention relates to a method for identifying the goat grass chromosome in the wheat genome, and belongs to the field of cytogenetics. Background technique [0002] The genus Goatwort contains 23 species, of which 11 are diploid and the remaining 12 are polyploid. The basic genome of species in this genus includes C, D, U, S, M, N and T. Genome T is only present in diploid species, and the rest of the genomes are present in both diploid and polyploid species. The species of this genus are divided by plant taxonomists into 6 distinct subgroups: Polyeides , Comopyrum , Cylindropyrum , Sitopsis , Vertebrata and Amblyopyrum . The last subgroup contains only Goatweed ( Ae. mutica, Also make Amblyopyrum muticum ) of a species, describing the uniqueness of that species. The uniqueness of goat grass is not only due to its unique spikelet characteristics at the filling stage (Ohta S, Saruhashi Y. Geographical distribution of B chromosome...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6841C12Q1/6895C12Q2563/107
Inventor 刘成宫文萍宫文英刘建军李根英韩冉宋健民李豪圣刘爱峰曹新有楚秀生黄承彦赵振东
Owner CROP RES INST SHANDONG ACAD OF AGRI SCI
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