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Method for identifying cymbidium varieties

A variety and sequence technology, applied in the field of plant variety identification, can solve the problems of poor specificity and small number of varieties, and achieve the effect of short time, fast speed and DNA fingerprint identification.

Inactive Publication Date: 2010-09-01
ZHEJIANG UNIV
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  • Abstract
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Problems solved by technology

[0003] There have been reports on the genetic polymorphism of Chunlan varieties in China, but the number of varieties is small, and random primers are mostly used, and the specificity is relatively poor

Method used

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  • Method for identifying cymbidium varieties
  • Method for identifying cymbidium varieties
  • Method for identifying cymbidium varieties

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Extract the genomic DNA of leaves of existing 112 Chunlan varieties; (2) carry out polymerase chain reaction (PCR reaction) respectively with the extracted Chunlan varieties genomic DNA to obtain simple repeat sequence products; (3) use 6% denaturation The polyacrylamide gel was used for electrophoresis separation of simple repetitive sequence products. The total loading volume was 5 μL, of which 4 μL was the amplification product, plus 1 μL Loading Buffer, and electrophoresed at a constant power of 60 W until the marker ran to 2 / 3 of the gel plate; After electrophoresis, take pictures with a digital camera by the Bassam silver staining method, and obtain the standard spectrum of simple repeat sequences respectively; (4) select a clear simple repeat sequence spectrum, use the "0-1" system to record the bands, and assign the bands to 1, the assignment of no band is 0; (5) read the data of the electrophoretic bands of the products amplified by different primers for eac...

Embodiment 2

[0031] The 'Laolongzi' variety ( figure 2 The Chinese code is LLZ) for identification, through the combination of 13 pairs of primers, combined with the calculation of the similarity coefficient, it was included in the standard map database for cluster analysis, and it was found that it was similar to the 'Longzi' variety ( figure 2The Chinese code is LZ) tightly combined. Statistical tests were performed on the fingerprints of LZ and LLZ, and the P value of the paired chi-square test of the contingency table was 1, indicating that the two varieties had the same fingerprints. The P value of the Kappa test is 0, so the null hypothesis is rejected and the two varieties are considered to be consistent. Combined with the Kappa index of 0.958, it can be considered that there is a very high consistency between them. Finally, it can be judged as the 'Dragon Character' variety. In conclusion, using one or more effective primers can distinguish and identify all Chunlan varieties in...

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Abstract

The invention provides a molecular identification method for cymbidium varieties, which is a simple sequence repeat genome analysis-based molecular identification method for the cymbidium varieties and analyzes the genetic relation of the cymbidium varieties by optimizing a technical system of cymbidium simple sequence repeat (SSR) analysis. By directly detecting the DNA sequence difference of the variety with genetic stability and information diversity, the method can definitely distinguish and judge different varieties of cymbidium, and realize accurate and quick identification of the variety so as to provide a reliable technique for variety identification, seed and seedling market management, variety protection and elite seed breeding and the like. By optimizing the SSR technical system of cymbidium genome DNA, the molecular identification of the cymbidium varieties has high speed, short time and multiple polymorphisms, the problems of reality inspection and the like of the cymbidium seedlings can be solved before trading, and the loss of a producer or a buyer is avoided. The method can distinguish and identify all cymbidium varieties by using one and more effective primers, and realize DNA fingerprint identification of the cymbidium varieties.

Description

technical field [0001] The invention belongs to a plant variety identification method, relates to a plant genetic marker and a variety identification method, and is a molecular identification method for Chunlan varieties based on genome SSR genetic analysis technology. Background technique [0002] Chunlan (Cymbidium goeringii) is a perennial herb plant, one of the traditional national orchids, native to China, with a long history of cultivation and breeding. Chunlan includes at least about 200 traditional varieties. It is one of the most expensive potted flowers in China, with high ornamental value and cultural value. In the process of genetic improvement of varieties, correctly dividing the variety groups and clarifying the genetic relationship between varieties has guiding significance for the selection and matching of parents, and is also one of the keys to the success of breeding work. The traditional classification of Chunlan varieties is mainly based on the petal sha...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 李方黄焱夏宜平陈昆松
Owner ZHEJIANG UNIV
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