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SRAP (Sequence-Related Amplified Polymorphism) molecular marking method related to color character of Ulmus pumila cv.jinye. and application thereof

A Chinese golden leaf elm, molecular marker technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., to achieve accurate results, guaranteed accuracy, and simple operation.

Inactive Publication Date: 2014-08-20
DAQING BRANCH OF HEILONGJIANG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no cDNA-SRAP technology has been applied in the screening analysis of differentially expressed genes in leaf color

Method used

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  • SRAP (Sequence-Related Amplified Polymorphism) molecular marking method related to color character of Ulmus pumila cv.jinye. and application thereof
  • SRAP (Sequence-Related Amplified Polymorphism) molecular marking method related to color character of Ulmus pumila cv.jinye. and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Primer screening of SRAP molecular markers related to leaf color traits of Ulmus chinensis

[0030] The Ulmus sinensis used in this example was purchased commercially.

[0031] 1. Leaf collection and leaf color calibration of different leaf colors of Ulmus sinensis:

[0032] Select 10 healthy and healthy Ulmus sinensis plants of the same tree age, respectively numbered 1 to 10, the leaf color calibration adopts the international color standard color palette comparison, select healthy young leaves with the same color value, and collect #FFD700 golden leaves from each plant and #228B22 forest green leaves, immediately sealed and stored at low temperature;

[0033] 2. Extract the total RNA from #FFD700 golden leaves and #228B22 forest green leaves on each Ulmus sinensis plant respectively. The extraction method of total RNA adopts the modified Trizol method, and the specific method is as follows:

[0034] A. Take 0.2g of Ulmus chinensis leaves, remove the peti...

Embodiment 2

[0059] Example 2: According to the SRAP molecular marker system related to leaf color traits of Ulmus chinensis in Example 1, identify Ulmus chinensis with different leaf colors.

[0060] figure 1 It is the amplification result of the two traits of yellow leaves and green leaves by the primer pair Me15 / Em19, wherein Y1-Y5 is a yellow leaf plant, YMix is ​​a mixed cDNA pool composed of Y1-Y5, G1-G4 is a green leaf plant, GMix is ​​a mixed cDNA pool composed of G1-G4. The two kinds of leaf colors can be well distinguished by the SRAP molecular marker method of the present invention. The method of the invention has good repeatability, good polymorphism, clear bands and low background.

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Abstract

The invention discloses an SRAP (Sequence-Related Amplified Polymorphism) molecular marking method related to color character of Ulmus pumila cv.jinye. and an application thereof, and relates to an SRAP molecular marking method and an application thereof. The method comprises the following steps: I, collecting leaves of different colors of Ulmus pumila cv.jinye. and calibrating the colors; II, respectively extracting total RNAs (Ribonucleic Acid) of golden and forest green leaves on each Ulmus pumila cv.jinye.; III, detecting RNA concentration; IV, carrying out cDNA-SRAP (Complementary Deoxyribonucleic Acid) amplification on differential fragments; and V, recovering, sequencing and analyzing the differential fragments. The method provided by the invention has the characteristics of simplicity in operation, accuracy in result, visualized result, practicality and the like. The method provided by the invention is used for researching genes related to different colors of Ulmus pumila cv.jinye.

Description

technical field [0001] The invention relates to a SRAP molecular marking method and application. Background technique [0002] In landscaping, colorful-leaved tree species have the characteristics of bright colors, long viewing period, and easy to form large color block landscapes. Their rich landscape levels make up for the lack of monotonous colors and lack of seasonal changes in cities. Various countries have paid more attention to the application of colorful leaf plants in gardens. Over the past hundred years, foreign developed countries have done a lot of work in the breeding and cultivation of colorful leaf plant varieties. In Germany, there are 64 species of ever-colored leaf trees used in urban gardens, which belong to 17 species, 35 subspecies or hybrids. They are colorful and properly configured to create a beautiful garden landscape effect. Domestic work on the breeding and introduction of color-leaved plants started relatively late and is still in its infancy. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王云云张兴高宇孙力肖宇关向军
Owner DAQING BRANCH OF HEILONGJIANG ACAD OF SCI
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