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A srap molecular marker primer combination and analysis method for genetic diversity analysis of P.

A technology of genetic diversity and molecular markers, applied in the field of SRAP molecular marker primer combination and analysis, to achieve high polymorphism, strong specificity, and uniform distribution

Active Publication Date: 2020-10-30
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The marker has the characteristics of economy, easy operation, high polymorphism, and good stability. It has been applied in many plants such as wheat, rice, Arabidopsis, cotton, and potato, but it has not been reported in the genus Papyrus.

Method used

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  • A srap molecular marker primer combination and analysis method for genetic diversity analysis of P.
  • A srap molecular marker primer combination and analysis method for genetic diversity analysis of P.
  • A srap molecular marker primer combination and analysis method for genetic diversity analysis of P.

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Of the 33 samples of Chonglou used in the present invention, 32 were collected from 6 areas in Jinzhai County, Anhui Province, and 1 was collected from Mount Huangshan. See Table 1 for details.

[0084] Table 1 Test material number and sampling point information

[0085]

[0086]

Embodiment 2

[0088] Extraction of Genomic DNA of Chonglou

[0089] The present invention collects the young leaves of Chonglou in Example 1 for the extraction of whole genome DNA, uses the CTAB method to extract the total genomic DNA of Chonglou, and dissolves the extracted DNA sample in TE buffer and places it at -20°C for later use. Before amplification, the DNA sample was diluted to 10 ng / μl with double distilled water, which was used as a template for PCR amplification reaction.

[0090] PCR reaction system and primers for SRAP marker

[0091] After seven pairs of primers, me2-em10, me3-em5, me3-em7, me3-em9, me3-em10, me4-em1 and me4-em4, each pair of primers was tested three times, the SRAP-PCR 25μl reaction system 10x PCR buffer (without Mg2+) 3.0μl, Mg2+1.5mmol / L, template DNA 40ng, dNTPs 0.25mmol / L, primer 0.4μmol / L, Taq enzyme 1U, the comprehensive amplification effect is the best. The PCR amplification reaction program was: 94°C pre-denaturation for 5 min, 94°C denaturation fo...

Embodiment 3

[0100] results and analysis

[0101] (1) Polymorphism analysis

[0102] Figure 1-7 It is the electrophoresis diagram of the amplification products of 33 samples of double building with 7 pairs of primer combinations. It can be seen from the figure that the obtained bands are clear and abundant. The primer amplification results are shown in Table 3. A total of 101 bands were amplified by 7 pairs of primer combinations, including 97 polymorphic bands, and the polymorphic percentage (PPB) was 96.04%. The bands amplified by different primers are different, and the bands amplified by the same primers and different regions are also different, indicating that there are abundant polymorphisms and complex genetic backgrounds among the 33 samples. Relationships offer possibilities.

[0103] Table 3 Primer amplification results

[0104]

[0105]

[0106] Note: TNP: total number of amplified bands; NPB: number of polymorphic bands; PPB: percentage of polymorphism; PIC: informat...

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Abstract

The invention provides an SRAP molecular marker primer composition for rhizoma paridis resource genetic diversity analysis and an analysis method and relates to the field of molecular biology DNA marking techniques and application techniques. The SRAP molecular marker primer composition for rhizoma paridis resource genetic diversity analysis, which is provided by the invention, comprises the following seven pairs of primers: me2-em10, me3-em5, me3-em7, me3-em9, me3-em10, me4-em1 and me4-em4. According to the method provided by the invention, the identification speed of rhizoma paridis resources can be increased, the testing time can be shortened, results are stable and reliable, and the defect that the conventional morphology identification method has large experiment errors can be avoided.

Description

technical field [0001] The invention relates to the field of molecular biology DNA marker technology and application technology, in particular to a combination of SRAP molecular marker primers and an analysis method for genetic diversity analysis of S. sinensis resources. Background technique [0002] Paris polyphylla is a perennial herbaceous plant of the genus Paris L. in the family Liliaceae. It mostly grows in damp places under forests, valleys or grass at an altitude of 600-3600 meters. It is slightly cold in nature, bitter in taste, and slightly poisonous. The rhizome is used as medicine, which has the effects of reducing swelling and pain, clearing heat and detoxification, calming shock and cooling the liver. Its main chemical components are steroidal saponins, which have biological activities such as antibacterial, anti-inflammatory, anti-tumor, hemostatic, sedative, and analgesic. In addition, Chonglou is also the main raw material of famous Chinese patent medicine...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 何金铃赵小培邹高芬赵洁胡林翼兰跃峰
Owner ANHUI AGRICULTURAL UNIVERSITY
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