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Two black mustard repeated DNA sequences and application thereof

A technology of DNA molecules and sequences, which is applied to two kinds of black mustard genome DNA sequences and its application fields, can solve the problems of cumbersome methods, poor repeatability, and the identification is greatly affected by subjective factors, and achieve the effect of easy identification

Active Publication Date: 2017-07-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 repeated DNA sequences and application thereof
  • Two black mustard repeated DNA sequences and application thereof
  • Two black mustard repeated DNA sequences and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the isolation of DNA repeat sequence BNSAT28 and BNSAT68

[0037] 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 BNSAT28 and BNSAT68 with abundant genome content were found, which were verified by PCR cloning.

[0038] The nucleotide sequence of BNSAT28 is sequence 1 in the sequence listing.

[0039] The nucleotide sequence of BNSAT68 is sequence 2 in the sequence listing.

Embodiment 2

[0040] Embodiment 2, the application of DNA repeat sequence BNSAT28 and BNSAT68 as detection probe

[0041] One, BNSAT28 and BNSAT68 as the fluorescence in situ hybridization (FISH) test method of detection probe

[0042] 1. Chromosome production

[0043] 1) Put the plant seeds to be tested in a petri dish with moist filter paper, and take root at room temperature;

[0044] 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;

[0045] 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;

[0046] 4) Excise part of the root tip meristem and place it in a mixed solution of 4% (volume percentage) cellulase and 1% (volume percentage) pectinase, and treat at 37°C for 50 minutes to soften the root tip;

[0047] 5) Apply the ...

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PUM

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Abstract

The invention discloses two black mustard genome DNA sequences and application thereof, and provides a DNA molecule set used for identifying a B genome of brassica plants. The DNA molecule set is composed of a single stranded DNA molecule as shown in sequence 1 in a sequence table and a single stranded DNA molecule as shown in sequence 2 in the sequence table. Experiments prove that by obtaining BNSAT28 and BNSAT68, chromosomes of brassica plants can be identified more easily and more accurately, and the problem that no ideal probe is available for identification of chromosomes of brassica plants is solved.

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 of black mustard, Brassica nigra (BB, n=8), and the C genome Brassica oleracea represented by cabbage (CC, n=9), and three heterotetraploid compound 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. Contains the world's most important edible, oily and seasoning vegetables, and is also a model material for studying genetic diversity within the genus and the evolution of species and polyploidy. [0003] Black mustard is a wild species of Brassica, which retains many excellent traits after long-term ecological adaptation, such as resistance ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/63
CPCC12Q1/6841C12Q1/6895C12Q2600/166C12Q2525/151
Inventor 王桂香刘凡赵泓何群燕宗梅韩硕郭宁李静蔡泽熙
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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