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Method for constructing fingerprint of phaseolus vulgaris according to SSR (simple sequence repeat) molecular marker and application

A technology of fingerprints and molecular markers, applied in biochemical equipment and methods, microbiological measurement/inspection, DNA preparation, etc., can solve the problems of distinguishing different bean varieties, different classification and naming standards of bean varieties, and achieve Short time consumption, accurate and reliable identification results, and effects that are not affected by the environment

Active Publication Date: 2016-03-23
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the classification and naming standards of kidney bean varieties are not uniform, and it is difficult to distinguish different kidney bean varieties from a single morphological point of view
There is no report on the establishment of DNA fingerprints of common bean cultivars to identify the cultivars

Method used

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  • Method for constructing fingerprint of phaseolus vulgaris according to SSR (simple sequence repeat) molecular marker and application
  • Method for constructing fingerprint of phaseolus vulgaris according to SSR (simple sequence repeat) molecular marker and application
  • Method for constructing fingerprint of phaseolus vulgaris according to SSR (simple sequence repeat) molecular marker and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] In this example, 36 kidney bean varieties were used as sample materials, and the fingerprint data of kidney bean varieties were constructed using the screened SSR core primers, so as to achieve the purpose of variety identification.

[0032] The information on the 36 tested kidney bean varieties is shown in Table 1 below. The germplasm numbers in Table 1 are from the China Crop Germplasm Information Network.

[0033] Table 136 information on the tested kidney bean varieties

[0034]

[0035] The construction of the standard DNA fingerprint library of kidney bean varieties is carried out according to the following steps:

[0036] 1. Extraction of total DNA from each variety of kidney bean

[0037] The bean was planted in the soil and cultured at room temperature. After 12 days of growth, the young bean leaves were taken and stored at -80°C after being frozen in liquid nitrogen. Take 0.2 g of tender young leaves of various varieties of kidney beans, put them in a 1....

Embodiment 2

[0060] Example 2 Primer polymorphism verification

[0061] 40 pairs of primers except the core primers were used to amplify 36 samples of kidney bean by PCR to construct the fingerprint of kidney bean. Statistical primer polymorphism information, the results of which 8 pairs of non-core primers are listed below are the fingerprints of 6 varieties. The information of 8 pairs of non-core primers is shown in Table 6, and the fingerprint codes of 5 tested varieties are shown in Table 7. Shown, fingerprint code primer sequence CD98, CD1, CD67, CD85, CD97, CD20, CD5, CG1

[0062] Table 5 Partial non-core primer sequences

[0063]

[0064]

[0065] Table 6 Non-core primer information

[0066] Primer name number of alleles PIC CD98 1 0 CD1 2 0.0912 CD67 2 0.121 CD85 1 0 CD97 1 0 CD20 1 0 CD5 1 0 CG1 2 0.151

[0067] Table 75 Fingerprint Codes of Varieties

[0068] breed name fingerprint co...

Embodiment 3

[0071] Take 10 kidney bean varieties (different from the place of origin in Example 1) as verification materials as shown in Table 8, use the core primers screened to amplify 10 kidney beans, and use the method in Example 1 to construct fingerprints for anonymous samples. The code is shown in Table 8.

[0072] Table 810 Verified Bean Varieties

[0073]

[0074]

[0075] Table 910 Species Fingerprint Codes

[0076] Numbering breed name fingerprint code 1 yellow kidney beans 4-3-3-2-2-2-3-1 2 Noodles 2-1-4-1-3-5-2-3 3 flower kidney beans 2-3-4-2-3-2-2-1 4 flower kidney beans 1-4-1-2-3-2-3-1 5 plum beans 1-4-3-2-2-4-3-3 6 big horseshoe 2-2-2-4-2-5-2-2 7 hard shell beans 2-3-1-1-2-4-1-1 8 Flower face beans 1-2-5-4-3-4-2-4 9 Milky Kidney Beans 1-1-2-3-1-5-1-4 10 Flower face beans 1-4-4-3-1-1-1-1

[0077] It can be seen from Table 8 and Table 3 that plum beans and eyebrow beans in Example 1...

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Abstract

The invention discloses a method for constructing a fingerprint of phaseolus vulgaris according to an SSR (simple sequence repeat) molecular marker and applications, and belongs to the technical field of finger print construction. The method comprises the following steps of extracting phaseolus vulgaris DNA (deoxyribonucleic acid), performing PCR (polymerase chain reaction) amplified reaction by utilizing the phaseolus vulgaris DNA and SSR core primers, and after electrophoresis detection, constructing the characteristic fingerprint of the phaseolus vulgaris according to an electrophoresis result; the nucleotide sequences of the SSR core primers are as shown by SEQIDNO.1-16. Compared with conventional morphological identification, the method disclosed by the invention is accurate and reliable in identification result and short in consumed time, is not influenced by environments, is simple in operation, can be used for identifying multiple varieties simultaneously, and is convenient and efficient.

Description

technical field [0001] The invention relates to a method and application for constructing a kidney bean fingerprint based on SSR molecular markers, and belongs to the technical field of fingerprint construction. Background technique [0002] Phaseolus vulgaris is one of the most important edible beans, accounting for 50% of the world's edible bean production. my country's phaseolus vulgaris is rich in germplasm and various in variety. After long-term selection and cultivation, different types of variety resources have been created. At present, the classification and naming standards of kidney bean varieties are not uniform, and it is difficult to distinguish different kidney bean varieties from a single morphological point of view. [0003] DNA fingerprinting refers to the DNA electrophoresis pattern that can identify the differences between biological individuals. It is an identification technology based on DNA molecular markers. The construction of DNA fingerprints plays ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/10
CPCC12N15/1003C12Q1/6895C12Q2525/151
Inventor 张东杰王颖杨义杰沈琰孙大庆赵雅楠
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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