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Method for identifying exogenous chromosomes and chromosome segments in plant distant hybrids

A technology of chromosome fragments and chromosome painting, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of unsuccessful detection, etc., and achieve the effect of clear fluorescent hybridization signal, strong specificity and strong signal

Inactive Publication Date: 2013-01-16
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chromosome painting techniques have not been successful for the detection of plant autosomes (ie, A chromosomes)

Method used

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  • Method for identifying exogenous chromosomes and chromosome segments in plant distant hybrids
  • Method for identifying exogenous chromosomes and chromosome segments in plant distant hybrids
  • Method for identifying exogenous chromosomes and chromosome segments in plant distant hybrids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1. Microseparation of rye 1R chromosome and identification of additional chromosomes and chromosome fragments derived from wheat-rye addition line 1R / CS and wheat-rye translocation line 1RS / 1BL rye

[0043] 1. Preparation of Rye Mitotic Metaphase

[0044] The rye (Secale cereal) king II used was purchased from the Pasture and Livestock Research Laboratory of the United States Department of Agriculture (USDA, ARS, FRRL, UT, USA).

[0045] 1)N 2 O: Use tweezers to pierce a hole in the cover of the 1.5ml centrifuge tube, open the cover, spray water into the centrifuge tube with a spray bottle until there is a layer of water droplets on the tube wall, cut out a 1-2cm root tip and put it into the tube. Put the N 2 Treat in O air chamber for 1-3h, the pressure is 10ATM (1.01Mpa);

[0046] 2) Fix the root tip with 90% acetic acid on ice for 10 minutes (no more than 1 hour) (you can directly add 90% acetic acid to the original 1.5ml tube with holes, or transfer the r...

Embodiment 2

[0083] Example 2. Microseparation of TAI-27 additional chromosomes and identification of TAI-27 additional chromosomes by chromosome painting

[0084] TAI-27, the allotropic addition line TAI-27, is the addition of two sets of 14 pairs of chromosomes from the two chromosome groups of Thipotinum intermedia (also known as Aureus aeruginosa) to two sets of allotropic addition lines of wheatgrass. One of the departments was purchased from Northeast Normal University.

[0085] 1. Preparation of TAI-27 Mitotic Metaphase

[0086] With embodiment 1.1.

[0087] 2. Microsegmentation of the additional Thiopyrum intermedia minichromosome in TAI-27

[0088] Firstly, a fine glass rod with a diameter of 1.0 mm is manually drawn into a fine glass needle with a diameter of about 1 μm by using a Leitz needle puller under an alcohol lamp. Under an inverted microscope (Olympus1M), use a hand-drawn glass needle with the aid of a Leitz micromanipulator (code-No. 93314, purchased from Leica, Germ...

Embodiment 37

[0109] Example 3. Microseparation of 7ES / CS additional Chromosome elongatum chromosome and its chromosome painting identification of 7ES / CS additional Chromoside elongatum chromosome

[0110] Materials used The wheat-Ethiopia elongatum isoaddition line 7ES was purchased from the Weizmann Institute of Sciences (Rehovot, Israel).

[0111] 1.7 Preparation of ES / CS Mitotic Metaphase

[0112] With embodiment 1.1.

[0113] 2.7 Microsegmentation of the Additional Minor Chromosome in Echinopsis elongatum in ES / CS

[0114] Firstly, a fine glass rod with a diameter of 1.0 mm is manually drawn into a fine glass needle with a diameter of about 1 μm by using a Leitz needle puller under an alcohol lamp. Under an inverted microscope (Olympus1M), use a hand-drawn glass needle with the help of a Leitz micromanipulator (code-No. 93314, purchased from Leica, Germany) to scratch the identified chromosomes back and forth, left and right, and scrape gently. Adhered to the small chromosomes of E....

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Abstract

The invention provides a method for identifying exogenous chromosomes and chromosome segments in plant distant hybrids. The method includes the steps of firstly, microscopically separating monosomes or chromosome segments of chromosomes to be tested of a plant to be tested; secondly, using the separated monosomes or chromosome segments in the first step to build a fluorescence probe of the monosomes or chromosome segments; thirdly, using the fluorescence probe built in the second step to perform fluorescence in situ hybridization to chromosomes in metaphase division of the hybrids; and fourthly, analyzing the fluorescence in situ hybridization result obtained in the third step to determine distribution of fluorescence signals of the chromosomes to be tested of the plant to be tested. Exogenous chromosomes and chromosome segments in wheat distant hybrids can be identified by the method, and the method is applicable to identification of exogenous chromosomes and chromosome segments in distant hybrids of other plants.

Description

technical field [0001] The invention relates to the establishment of a plant chromosome painting technique and its application in the identification of exogenous chromosomes and chromosome fragments in plant distant hybrids. Specifically, the present invention relates to a method for identifying exogenous chromosomes and chromosome fragments of plant distant hybrids using chromosome painting technology. Background technique [0002] Wheat (Triticum aestirum L.) (2n=6x=AABBDD) is one of the most important food crops in the world, and its planting area and output rank first among cereal crops. In my country, wheat is second only to rice and occupies a very important position in agriculture. The genetic basis of Chinese wheat varieties is relatively narrow, which largely limits the improvement of wheat yield and quality. The relative species of wheat is a huge and precious gene resource bank, with rich genetic variation and many excellent characteristics that cultivated wheat...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 胡赞民邓传良符书兰韩方普尹维波陈宇红
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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