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Method for identifying red bean varieties by utilizing genome SSR and EST-SSR fingerprints and application

A technology of adzuki bean varieties, applied in the fields of molecular biology and plant variety identification, can solve the problems of homonym and synonym of adzuki bean varieties and high genetic similarity of varieties

Inactive Publication Date: 2016-12-07
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

[0003] Due to the introduction and utilization of different regions, the homonym phenomenon of adzuki bean varieties is very serious, and the genetic similarity of local varieties is high. It is difficult for traditional morphological and cytological identification methods to accurately and truly identify the differences in adzuki bean varieties. Analysis and identification at the molecular level

Method used

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  • Method for identifying red bean varieties by utilizing genome SSR and EST-SSR fingerprints and application
  • Method for identifying red bean varieties by utilizing genome SSR and EST-SSR fingerprints and application
  • Method for identifying red bean varieties by utilizing genome SSR and EST-SSR fingerprints and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 is used to identify the screening of SSR and EST-SSR primers of Adzuki bean varieties

[0034] In this example, the genome DNA of 32 adzuki bean varieties (from the National Crop Germplasm Resources Preservation Center) representatively popularized in the main production areas of adzuki bean in my country was used as a template, and the core genome SSR and EST-SSR were synthesized by analyzing the adzuki bean genome sequence and EST sequence primers. After screening, the molecular marker combinations composed of 5 pairs of genomic SSR and 5 pairs of EST-SSR primers can completely identify the differences of 32 adzuki bean varieties.

[0035] The 32 representative adzuki bean varieties are: Nenlu 1, Bao 942-40-2, An 9910, Xinglv 1, An 05-4, Bao 9815-36, Wei 8901-32, Zhenglv 8, Yulu No. 1, No. 1 Elv, No. 4 Yulv, No. 3 Zhonglv, Bao 861-10-6, No. 1 Jilv, No. 5 Zhonglv, No. 6 Bailv, No. 1 Anheilu, Bao 865-18 -9, Jilv 4, Jilv 9346, Jilv 9239, Jinxiaodou 2, Liaolv 6...

Embodiment 2

[0043] Example 2 Construction of Adzuki Bean Variety Characteristic Fingerprint and Establishment of Adzuki Bean Variety Identification Method

[0044] 1. Total DNA extraction of adzuki bean varieties

[0045] The adzuki bean was planted in the soil and cultured at room temperature. After two weeks of growth, the young adzuki bean leaves were frozen in liquid nitrogen and stored at -80°C for later use. Take 0.2 g of tender young leaves of adzuki bean of each variety in a 1.5 mL centrifuge tube, add liquid nitrogen and 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 quality 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.

[0046] 2. Taking 32 parts of adzuki bean varieties described in Example 1 as materials, use the 5 pairs of genome SSR molecular m...

Embodiment 3

[0073] Embodiment 3 The application of the present invention's identification method for adzuki bean varieties

[0074] DNA was extracted from leaves of Jingnong 2 and Jihongxiaodou 3. The 10 pairs of primers in the molecular marker combination screened in Example 1 of the present invention are used to perform PCR amplification on the DNA of the adzuki bean species to be checked respectively; The method of electrophoresis and strip recording, compared the obtained fingerprint codes with the characteristic fingerprints of the above-mentioned adzuki bean varieties, and found that the fingerprint code of Jingnong No. 2 was 0110101010011010101, and the fingerprint code of Jihong Xiaodou No. 2 The fingerprint codes of Jingnong No. 2 and Jihongxiaodou No. 3 are the same, indicating that the method for identifying adzuki bean varieties established in Example 2 of the present invention is more accurate in verifying adzuki bean varieties and is suitable for popularization.

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Abstract

The invention provides a method for identifying red bean varieties by utilizing genome SSR and EST-SSR fingerprints and application and belongs to the technical field of plant variety identification. A mark combination composed of 5 pairs of genome SSRs and 5 pairs of EST-SSR primers is provided, nucleotide sequences of the 5 pairs of genome SSRs and the 5 pairs of EST-SSR primers are respectively as shown in SEQ ID No. 1-20, and fingerprints of 32 kinds of red beans are constructed; according to the fingerprints, red bean seed variety identification and genetic diversity evaluation work can be finished within 4 hours by adopting the mark combination, and the method has the advantages of being efficient, accurate, low in cost and simple and convenient to operate. According to the method, red bean seed authenticity is effectively monitored, variety heritable variation and genetic relationships of varieties are disclosed from the DNA level, crop varieties are protected, forged and fake commodity varieties are prevented from entering the market, technical support is provided for reasonable utilization of excellent germ plasm in the red bean variety breeding process, and the method has good application prospect.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and plant variety identification, in particular to a method and application for identifying adzuki bean varieties by using genome SSR and EST-SSR fingerprints. Background technique [0002] Adzuki bean (Vigna angularis), belonging to the leguminous family, has been cultivated in China for two thousand years. It is an important food crop in our country. Because of its important nutritional value and medical value, it has been developed as an important functional food. In recent years, with the adjustment of the planting industry structure and the change of people's dietary structure, people's demand for adzuki beans is increasing, and the planting area is expanding year by year. Studies have shown that pests and diseases seriously affect the germination and yield of adzuki bean seeds. In view of this, in-depth study of the genes involved in insect resistance and the mechanism of insec...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 陈红霖程须珍王丽侠王素华
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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