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Avena L. DNA (deoxyribonucleic acid) molecular fingerprint spectrum construction method and application

A technology of DNA molecules and fingerprints, which is applied in the field of molecular biology and plant variety identification, can solve the problem of high development costs and achieve the effect of preventing counterfeit and inferior varieties from entering the market

Inactive Publication Date: 2018-01-09
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the discovery of SSR markers needs to go through the steps of cloning, sequencing, artificially designing and synthesizing primers, and screening the flanking sequences of microsatellites, so the development cost is quite high. There is no report on the establishment of DNA fingerprints of oat varieties to identify varieties.

Method used

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  • Avena L. DNA (deoxyribonucleic acid) molecular fingerprint spectrum construction method and application
  • Avena L. DNA (deoxyribonucleic acid) molecular fingerprint spectrum construction method and application
  • Avena L. DNA (deoxyribonucleic acid) molecular fingerprint spectrum construction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 110

[0030] Example 1 Obtaining 10 pairs of core SSR primers

[0031] In this example, using the genomic DNA of 30 representative oat varieties (from the National Crop Germplasm Resources Preservation Center) in different production areas in my country as templates, by analyzing the number of SSR marker allelic sites, polymorphic information content values ​​and possible Repeatability and other indicators, 10 pairs of SSR primers were screened out, and the molecular marker combinations composed of these primers could completely identify the differences of 30 oat varieties.

[0032] The 30 representative oat varieties are: Pinyan No. 1, Ningyou No. 1, Longyan No. 2, Dingyou No. 1, Bayou No. 1, Bayou No. 9, Baiyan No. 2, Baiyan No. 7, Baiyan No. Yinyan No.1, Caoyou No.1, Huawan No.6, Huazao No.2, Zhangyan No.7, Keyan No.1, Pin No.5, Bayan No.4, Wuzhai Naked Oats, Lijiang Oats, Wuyan No.5 , North China No. 1, Zaoyan No. 1, Jiza No. 2, Hefeng No. 1, Jinyan No. 7, Yanhong No. 10, Tongxi...

Embodiment 2

[0037] Example 2 Construction of Oat Variety Characteristic Fingerprint and Establishment of Oat Variety Identification Method

[0038] 1. Genomic DNA extraction of oat varieties

[0039] The oats were planted in the soil and cultured at room temperature. After two weeks of growth, 0.2 g of the young leaves of each variety of oats were taken in a 1.5 mL centrifuge tube, and liquid nitrogen was added to grind them into powder. The DNA was extracted by the CTAB method, and the experimental steps were carried out according to the instructions of the kit. The concentration of the total DNA was detected by agarose gel electrophoresis and a spectrophotometer, and then the total DNA was diluted to 100 ng / μL and stored at -20°C for future use.

[0040] 2. Using the 30 oat varieties described in Example 1 as materials, use the 10 pairs of SSR primers obtained by screening to carry out PCR amplification of the DNA of each oat variety respectively. The PCR reaction system used in the PC...

Embodiment 3

[0051] Embodiment 3 Application of the method for identifying oat varieties of the present invention

[0052] Take 2 oat varieties (different from the origin in Example 1) as verification materials, use the screened core primers to amplify the 2 oats, and use the method in Example 2 to construct fingerprints for unknown samples. The obtained fingerprints are shown in the table 3.

[0053] Table 3 Fingerprints of varieties to be tested (variety 1 and variety 2)

[0054] name

[0055] Comparing the obtained fingerprint table 3 with the characteristic fingerprint table 2 of the above-mentioned known oat varieties, it is found that the fingerprint codes of the varieties to be tested are the same as those of "Bayou No. 1" and "Baiyan No. 2" in Table 2, indicating that are of the same species as each other.

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Abstract

The invention discloses an avena L. DNA (deoxyribonucleic acid) molecular fingerprint spectrum construction method and application. According to the method, a set of specific SSR (simple sequence repeats) molecular marker primers (SEQ ID NO: 1-20) are designed aiming at avena L. genomes, and can be used for performing PCR (polymerase chain reaction) amplification on avena L. genome DNAs to generate amplification products with different lengths, and the lengths of the products are analyzed and detected to construct characteristic fingerprint spectrums of different avena L. varieties to implement variety identification. Compared with conventional morphological identification, the variety identification method provided by the invention has the advantages of low time consumption, no environmental influence, convenience, rapidness, accurate and reliable result and broad application prospect.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and plant variety identification, in particular to a construction method and application of an oat DNA molecular fingerprint. Background technique [0002] Oat (Avena L.) is an annual herbaceous plant belonging to the Gramineae family. It is an important food and forage crop widely distributed all over the world. According to whether oat grains are shelled or not, oats can be divided into two categories: hulled oats and naked oats. The oats cultivated in Europe and the United States are mainly hulled oats, and the research is mainly on hulled oats; China is the origin center of large naked oats. Naked oats have a long history of cultivation in China, and so far naked oats are still mainly grown in major oat producing areas. Oats are rich in nutrients and are one of the best complete nutritional foods among grains. Compared with wheat, rice, corn and other crops, the nutritional indicat...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895
Inventor 吴斌
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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