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Two black mustard genomes DNA sequence and application thereof

A genome and DNA molecule technology, applied to two black mustard genome DNA sequences and their application fields, can solve the problems of cumbersome methods, the identification is greatly influenced by subjective factors, and the repeatability is poor, so as to achieve the effect of easy identification

Inactive Publication Date: 2016-10-26
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on the karyotype analysis of the B genome of Brassica, and it basically stays on the chromosome morphology and C banding technology, which has poor repeatability, and the identification is greatly affected by subjective factors, and the method is cumbersome

Method used

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  • Two black mustard genomes DNA sequence and application thereof
  • Two black mustard genomes DNA sequence and application thereof
  • Two black mustard genomes DNA sequence and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1, the separation of DNA repeat sequence CL13 and CL76

[0034] Using black mustard 'G1 / 1' as a material, the base number of 0.6G obtained by random genome sequencing is equivalent to about 0.8 times of the black mustard genome (calculated based on the genome size of 630Mb, Johnston et al., 2005). Combining the methods of Novák et al. (2010) and Macas et al. (2010) for repeat sequence analysis, two tandem repeat sequences CL13 and CL76 with abundant genome content were found, and verified by PCR cloning.

[0035] The nucleotide sequence of CL13 is sequence 1 in the sequence listing.

[0036] The nucleotide sequence of CL76 is sequence 2 in the sequence listing.

Embodiment 2

[0037] Embodiment 2, the application of DNA repeat sequence CL13 and CL76 as detection probe

[0038] 1. Fluorescence in situ hybridization (FISH) test method using CL13 and CL76 as detection probes

[0039] 1. Chromosome production

[0040] 1) Put the plant seeds to be tested in a petri dish with moist filter paper, and take root at room temperature;

[0041] 2) When the root tip length is 0.5-1cm, cut off the root tip and put it into the d 2 h 2 O in a test tube, and put the test tube into an ice box filled with ice-water mixture and place it in the refrigerator for 22-24h for pretreatment;

[0042] 3) Put the pretreated root tip into fixative solution (absolute ethanol:glacial acetic acid=3:1 (v / v)) and fix it for 1-7 days;

[0043] Note: In order to obtain better results in in situ hybridization, the fixation time should not be too long. Of course, if the fixation time is less than two days, it is relatively difficult to make slices.

[0044] 4) Excise part of the roo...

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Abstract

The invention discloses two black mustard genomes DNA sequence and application thereof. A set of DNA molecules used for identifying brassica plant B genome are composed of single-stranded DNA molecules as shown in the sequence of the sequence table and single-stranded DNA molecules as shown in the sequence 2 of the sequence table. It is proved through experiments that it is easier and more accurate to identify chromosome of brassica plants by the acquisition of CL13 and CL76. The gap that there is no ideal probe for identification of chromosome of brassica plants is filled.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to two genomic DNA sequences of black mustard and applications thereof. Background technique [0002] Cruciferous Brassica vegetables include the A genome Brassica.rapa (AA, n = 10) represented by Chinese cabbage, the B genome B of black mustard, B. nigra (BB, n = 8), and the C genome represented by cabbage B. oleracea (CC, n = 9), and three tetraploid complex species formed by their natural doubling: Brassica napus B. napus (AACC, n = 19), Brassica napus B. juncea (AABB, n=18), and B. carinata (BBCC, n=17), formed the famous U-triangle. Black mustard is a wild species of Brassica. Compared with the agriculturally important diploid species of cabbage and cabbage, the genetic and cytological studies of black mustard are relatively limited. The evolution and ploidy evolution of Brassica species are also affected by the B genome. Limited by lack of information. [0003] Black mustard ha...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6841C12Q1/6895C12Q2563/107
Inventor 王桂香刘凡何群燕李静宗梅郭宁韩硕
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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