SSR Molecular Markers of Codonopsis tangshen Oliv. and Their Applications, and Identification Method of Codonopsis tangshen Oliv. Medicinal Materials

Abstract

The present invention discloses an SSR molecular marker of Codonopsis pilosula, which is obtained by amplifying an upstream primer having a sequence as shown in SEQ ID NO: 1 and a downstream primer having a sequence as shown in SEQ ID NO: 2. The 5' end of the upstream primer is mounted with a fluorescent marker group, and the fluorescent marker group is selected from one of fluorescein FAM, fluorescein ROX, fluorescein HEX and fluorescein TAMARA. The present invention also discloses the application of the above-mentioned SSR molecular marker and a method for identifying Codonopsis pilosula. The present invention can effectively and specifically identify Codonopsis pilosula and Panthera pilosula medicinal materials from Codonopsis pilosula, Codonopsis pilosula, Codonopsis pilosula and Panthera pilosula medicinal materials.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine identification, in particular to an SSR molecular marker of Codonopsis tangshen Oliv., its application, and a method for identifying Codonopsis tangshen Oliv. medicinal materials. Background Art

[0002] Codonopsis pilosula is a medicinal material with multiple origins. The origins are respectively Codonopsis pilosula (Franch.) Nannf., Codonopsis pilosula Nannf. var. modesta (Nannf.) L.T. Shen, or Codonopsis tangshen Oliv. of the genus Codonopsis in the family Campanulaceae. It is mainly distributed in Lingchuan, Jincheng, Shanxi; Weiyuan, Dingxi, Gansu; Longnan, Gansu; Gangu, Tianshui, Gansu; Weining, Bijie, Guizhou and other places. Codonopsis pilosula has the effects of strengthening the spleen and benefiting the lung, nourishing blood and promoting fluid production, and is widely used and has a large dosage in clinical practice.

[0003] In recent years, with the gradual promotion of the principle of "one medicine, one name, one origin" for traditional Chinese medicine, the State Food and Drug Administration has issued the "Technical Requirements for Quality Control and Standard Setting of Traditional Chinese Medicine Formula Granules", which stipulates that "if the source is traditional Chinese medicinal materials with multiple origins, one origin should be fixed, and traditional Chinese medicinal materials with different origins cannot be mixed with each other". Judging from the appearance, the medicinal material forms of the three kinds of Codonopsis pilosula are extremely similar, and there is no other effective distinguishing method at present. In addition, in addition to different origins of Codonopsis pilosula, the roots of other plants of the same family are also used as Codonopsis pilosula for medicinal purposes and flow into the market in some local areas, but their effects are very different. Especially for Campanumoea javanica Bl. of the genus Campanumoea in the family Campanulaceae, its dried root is commonly known as "wild Codonopsis pilosula", "wild Codonopsis fruit", "wild ginseng", etc., and is used as "wild Codonopsis pilosula" in Guizhou and other regions. There are large interspecific variations and significant differences in medicinal effects between it and Codonopsis pilosula medicinal materials. Therefore, it is of great significance to establish a suitable method to distinguish Codonopsis pilosula medicinal materials with different origins and their formula granules.

[0004] Currently, the molecular identification techniques for Codonopsis pilosula mainly include DNA barcoding, SSR markers, ISSR markers, etc., which are mostly used for the study of species genetic diversity among Codonopsis species. However, due to the certain degree of confusion and impurity in the cultivation of Codonopsis pilosula, it is difficult to ensure the variety purity by appearance morphology and physical and chemical indexes. Currently, there are relatively few relevant literatures on SSR markers of Codonopsis pilosula. Relevant patents include that in the cpSSR markers of Codonopsis species, through 5 pairs of cpSSR markers and agarose electrophoresis, Codonopsis lanceolata can be distinguished from wild and cultivated Codonopsis pilosula; in a series of patents on Codonopsis lanceolata and Adenophora triphylla, SSR markers are mostly used to distinguish Codonopsis lanceolata, Codonopsis pilosula, Adenophora triphylla, Codonopsis ussuriensis and different ages of Codonopsis pilosula, etc.

[0005] However, the above-mentioned public information does not involve the identification of different origins and other counterfeit medicinal materials of medicinal Codonopsis pilosula, and it is impossible to effectively distinguish different origins of Codonopsis pilosula. Therefore, it is of great significance to identify the origin of Codonopsis tangshen Oliv. in Codonopsis pilosula by SSR molecular markers, and to specifically identify Codonopsis tangshen Oliv. medicinal materials, so as to fix the origin of Codonopsis pilosula from multiple sources and make it meet the medicinal standards. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide an SSR molecular marker for Codonopsis tangshen Oliv., which can effectively identify the authenticity of Codonopsis tangshen Oliv. medicinal materials.

[0007] Another technical problem to be solved by the present invention is to provide the application of the SSR molecular marker of Codonopsis tangshen Oliv.

[0008] Another technical problem to be solved by the present invention is to provide a method for identifying Codonopsis tangshen Oliv. medicinal materials, which has good accuracy and high specificity.

[0009] To solve the above technical problems, the present invention provides an SSR molecular marker for Codonopsis tangshen Oliv., and the SSR molecular marker for Codonopsis tangshen Oliv. is amplified by an upstream primer with a sequence as shown in SEQ ID NO: 1 and a downstream primer with a sequence as shown in SEQ ID NO: 2. Among them, a fluorescent labeling group is attached to the 5' end of the upstream primer, and the fluorescent labeling group is selected from one of fluorescein FAM, fluorescein ROX, fluorescein HEX and fluorescein TAMARA.

[0010] Table 1 shows the specific sequences of the upstream primer and the downstream primer

[0011]

[0012] To solve the above technical problems, the present invention provides the application of the SSR molecular marker of Codonopsis tangshen Oliv. in (1) or (2):

[0013] (1) Identifying the authenticity of Codonopsis tangshen Oliv.;

[0014] (2) Specifically identifying Codonopsis tangshen Oliv. and Campanumoea javanica Blume from Codonopsis pilosula (Franch.) Nannf., Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen, Codonopsis tangshen Oliv. and Campanumoea javanica Blume medicinal materials.

[0015] In one embodiment, it includes the following steps:

[0016] Extracting the genomic DNA of the sample to be tested;

[0017] Using the genomic DNA of the sample to be tested as a template, performing PCR amplification with the upstream primer and downstream primer of the SSR molecular marker of Codonopsis tangshen Oliv. as described in Table 1 to obtain a PCR amplification product;

[0018] Performing capillary electrophoresis detection on the PCR amplification product;

[0019] If characteristic double peaks of 105bp - 106bp and 106bp - 107bp fragments appear in the detection, the sample to be tested is Codonopsis tangshen Oliv. medicinal material;

[0020] If characteristic double peaks of 96bp - 97bp and 97bp - 98bp fragments appear in the detection, the sample to be tested is Campanumoea javanica Blume medicinal material.

[0021] To solve the above technical problems, the present invention provides a method for identifying Codonopsis tangshen Oliv. medicinal material, including the following steps:

[0022] Extracting the genomic DNA of the sample to be tested;

[0023] Using the genomic DNA of the sample to be tested as a template, performing PCR amplification with the upstream primer and downstream primer of the SSR molecular marker of Codonopsis tangshen Oliv. as described in Table 1 to obtain a PCR amplification product;

[0024] Performing capillary electrophoresis detection on the PCR amplification product, and judging whether the sample to be tested is Codonopsis tangshen Oliv. medicinal material according to the detection result.

[0025] In one embodiment, the program of the PCR amplification is: pre - denaturing the amplification system at a temperature of 94°C - 96°C for 2min - 4min;

[0026] Denaturing at a temperature of 94°C - 96°C for 5s - 15s, then annealing at a temperature of 50°C - 60°C for 5s - 15s, and then extending at a temperature of 70°C - 75°C for 5s - 15s, repeating the above operations 33 - 36 times;

[0027] After extension at a temperature of 70°C - 75°C for 4 min - 6 min, storage is completed at 3°C - 5°C.

[0028] In one embodiment, the reaction system for PCR amplification includes: 9 μL - 11 μL of 2×M5 Supper FastTaq PCR, 0.2 μL - 0.8 μL of the upstream primer, 0.2 μL - 0.8 μL of the downstream primer, 0.5 μL - 1.5 μL of DNA template, and sterilized double-distilled water to make up to 20 μL.

[0029] In one embodiment, the concentration of the DNA template is 25 ng / μl - 35 ng / μl;

[0030] The concentration of the upstream primer is 8 μmol / L - 12 μmol / L;

[0031] The concentration of the downstream primer is 8 μmol / L - 12 μmol / L.

[0032] In one embodiment, the electrophoresis detection includes: detecting the amplified sequence by capillary electrophoresis.

[0033] In one embodiment, if characteristic double peaks of 105 bp - 106 bp and 106 bp - 107 bp fragments appear in the detection result, the test sample is the medicinal material of Codonopsis tangshen Oliv.; otherwise, the test sample is a fake or inferior product.

[0034] In one embodiment, if characteristic double peaks of 96 bp - 97 bp and 97 bp - 98 bp fragments appear in the detection result, the test sample is the medicinal material of Campanumoea javanica Blume.

[0035] Implementing the present invention has the following beneficial effects:

[0036] The SSR molecular marker of Codonopsis tangshen Oliv. provided by the present invention can specifically identify the medicinal materials of Codonopsis tangshen Oliv. and Campanumoea javanica Blume from Codonopsis pilosula (Franch.) Nannf., Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen, Codonopsis tangshen Oliv. and Campanumoea javanica Blume.

[0037] The identification method of the medicinal material of Codonopsis tangshen Oliv. provided by the present invention can be used for specific identification of Codonopsis tangshen Oliv. It includes extracting genomic DNA, using the above genomic DNA as a template, performing PCR amplification with the designed primers, and detecting the amplified sequence by capillary electrophoresis. The allele size is determined with reference to the GS-LIZ500 molecular weight internal standard using GeneMarker. If characteristic double peaks of 105 bp - 106 bp and 106 bp - 107 bp fragments appear in the detection, the species is the medicinal material of Codonopsis tangshen Oliv.; if characteristic double peaks of 96 bp - 97 bp and 97 bp - 98 bp fragments appear, the species is the medicinal material of Campanumoea javanica Blume. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 Results of primer screening in Example 1 of the present invention; where M: 20 DNA Ladder; 1: Codonopsis tangshen Oliv.; 2: Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen; 3: Codonopsis pilosula (Franch.) Nannf.

[0039] Figure 2 Results of sample detection in Example 1 of the present invention. Detailed implementation manners

[0040] To make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below.

[0041] Unless otherwise specified or there are contradictions, the terms or phrases used herein have the following meanings:

[0042] In the present invention, "preferred" only describes the implementation manners or embodiments with better effects, and it should be understood that it does not constitute a limitation on the protection scope of the present invention.

[0043] In the present invention, among the technical features described in an open-ended manner, it includes a closed technical solution composed of the listed features, and also includes an open technical solution containing the listed features.

[0044] In the present invention, regarding the numerical range, unless otherwise specified, it includes both endpoints of the numerical range.

[0045] To make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and specific implementation manners.

[0046] To solve the above technical problems, the present invention provides an SSR molecular marker for Codonopsis tangshen Oliv., and the SSR molecular marker for Codonopsis tangshen Oliv. is amplified by an upstream primer with a sequence as shown in SEQ ID NO: 1 and a downstream primer with a sequence as shown in SEQ ID NO: 2. Among them, a fluorescent labeling group is attached to the 5' end of the upstream primer, and the fluorescent labeling group is selected from one of fluorescein Fam, fluorescein ROX, fluorescein HEX, and fluorescein TAMARA.

[0047] Table 1 shows the specific sequences of the upstream primer and the downstream primer

[0048]

[0049] Correspondingly, the present invention provides the application of the SSR molecular marker for Codonopsis tangshen Oliv. in (1) or (2):

[0050] (1) Identifying the authenticity of Codonopsis tangshen Oliv.;

[0051] (2) Specifically identifying Codonopsis tangshen Oliv. and Campanumoea javanica Blume from Codonopsis pilosula (Franch.) Nannf., Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen, Codonopsis tangshen Oliv., and Campanumoea javanica Blume.

[0052] In one embodiment, the application of the SSR molecular markers of Codonopsis tangshen Oliv. includes the following steps:

[0053] Extract the genomic DNA of the sample to be tested;

[0054] Using the genomic DNA of the sample to be tested as a template, perform PCR amplification with the upstream primer and downstream primer of the SSR molecular markers of Codonopsis tangshen Oliv. as described in Table 1 to obtain a PCR amplification product;

[0055] Perform electrophoresis detection on the PCR amplification product;

[0056] If characteristic double peaks of fragments of 105bp - 106bp and 106bp - 107bp appear in the detection, the sample to be tested is Codonopsis tangshen Oliv. medicinal material;

[0057] If characteristic double peaks of fragments of 96bp - 97bp and 97bp - 98bp appear in the detection, the sample to be tested is Campanumoea javanica Blume medicinal material.

[0058] Correspondingly, the present invention provides a method for identifying Codonopsis tangshen Oliv. medicinal material, including the following steps:

[0059] S1. Extract the genomic DNA of the sample to be tested;

[0060] In one embodiment, use the Wizard Genomic DNA Purification System extraction kit (A2361) (Promega, USA) to extract genomic DNA, use a Drop-nano micro ultraviolet spectrophotometer to measure the DNA concentration, record A260 / A280 and A260 / A230 at the same time, and adjust the concentration to 30 ng / μl. The adjusted sample is stored in a refrigerator at 4°C.

[0061] S2. Using the genomic DNA of the sample to be tested as a template, perform PCR amplification with the upstream primer and downstream primer of the SSR molecular markers of Codonopsis tangshen Oliv. as described in Table 1 to obtain a PCR amplification product;

[0062] In one embodiment, the procedure for the PCR amplification is as follows:

[0063] A. Pre-denature the amplification system at a temperature of 94°C - 96°C for 2 min - 4 min;

[0064] B. Denature at a temperature of 94°C - 96°C for 5 s - 15 s, then anneal at a temperature of 50°C - 60°C for 5 s - 15 s, and then extend at a temperature of 70°C - 75°C for 5 s - 15 s. Repeat the above operations 33 - 36 times;

[0065] C. Extend at a temperature of 70°C - 75°C for 4 min - 6 min and then complete the preservation at 3°C - 5°C.

[0066] In one embodiment, the reaction system for PCR amplification includes: 9 μL - 11 μL of 2×M5 Supper FastTaq PCR, 0.2 μL - 0.8 μL of the upstream primer, 0.2 μL - 0.8 μL of the downstream primer, 0.5 μL - 1.5 μL of DNA template, and sterilized double-distilled water to make up to 20 μL.

[0067] Preferably, the procedure for PCR amplification is as follows:

[0068] A. Pre-denature the amplification system at a temperature of 95°C for 3 min;

[0069] B. Denature at a temperature of 95°C for 10 s, then anneal at a temperature of 55°C for 10 s, and then extend at a temperature of 72°C for 10 s, and repeat the above operations 35 times;

[0070] C. Extend at a temperature of 72°C for 5 min and then complete the preservation at 4°C.

[0071] The reaction system for PCR amplification includes: 10 μL of 2×M5 Supper FastTaq PCR, 0.5 μL of the upstream primer, 0.5 μL of the downstream primer, 1 μL of DNA template, and sterilized double-distilled water to make up to 20 μL.

[0072] S3. Perform capillary electrophoresis detection on the PCR amplification product, and judge whether the test sample is Codonopsis tangshen Oliv. according to the detection result.

[0073] If characteristic double peaks of 105 bp - 106 bp and 106 bp - 107 bp fragments appear in the detection, the test sample is Codonopsis tangshen Oliv.;

[0074] If characteristic double peaks of 96 bp - 97 bp and 97 bp - 98 bp fragments appear in the detection, the test sample is Campanumoea javanica Blume.

[0075] The present invention will be described below with specific examples.

[0076] Example 1

[0077] The method for identifying Codonopsis tangshen Oliv. specifically includes the following steps:

[0078] S1. Extract the genomic DNA of the test sample;

[0079] Prepare the samples according to Table 2 and extract the genomic DNA of the samples according to the following method.

[0080] Table 2 is the sample list

[0081]

[0082]

[0083] Then, use the Wizard Genomic DNA Purification System extraction kit (A2361) (Promega, USA) to extract the genomic DNA of the above samples. Measure the DNA concentration using a Drop-nano micro ultraviolet spectrophotometer, and record A260 / A280 and A260 / A230 simultaneously, and adjust the concentration to 30 ng / μl. The adjusted samples are stored in a refrigerator at 4°C.

[0084] S2. Using the genomic DNA of the sample to be tested as a template, perform PCR amplification using the upstream and downstream primers of the SSR molecular marker of Codonopsis tangshen as described in Table 1 to obtain PCR amplification products;

[0085] 2.1 Primer design

[0086] Mine the SSR loci, design primers through primer 5.0, and the obtained sequences are shown in Table 3. The primers are synthesized by Shanghai Sangon Biological Engineering Co., Ltd.

[0087] Table 3 is the primer information table

[0088]

[0089] 2.2 PCR amplification

[0090] The PCR reaction system is as follows: 2×M5 Supper FastTaq PCR 10 μl, 0.5 μl each of the upstream and downstream primers (10 μM), 1 μl of template DNA (30 ng / μl), and sterilized water is added to make up to a 20 μl system.

[0091] The PCR reaction program: pre-denaturation at 95°C for 3 min; 35 cycles: denaturation at 95°C for 10 s, annealing at 55°C for 10 s, extension at 72°C for 10 s; then extension at 72°C for 5 min and preservation at 4°C.

[0092] S3. Detect the differential bands of the PCR amplification products by polyacrylamide gel electrophoresis, label the target sequence with a detectable fluorescent substance, detect the amplified sequence by capillary electrophoresis, and judge whether the sample to be tested is Codonopsis tangshen medicinal materials according to the detection results.

[0093] Under the conditions of a voltage of 130 V and a current of 20 mA, in 1×TBE buffer, through 10% polyacrylamide gel electrophoresis, using 20 DNA ladder and DL1000 Marker (Beijing Baori Biotechnology Co., Ltd., TAKARA) as references, the PCR products were detected by silver staining method, and the target sequences were labeled with detectable fluorescent substances, and the amplified sequences were detected by capillary electrophoresis. The results are as Figure 1 and Figure 2 and Table 4 show. From the above results, it can be seen that the primer combination, amplification system and detection method of the SSR molecular marker provided by the present invention can effectively identify Codonopsis tangshen Oliv., Codonopsis pilosula Nannf., Codonopsis sichuanensis (Pai) Hong and Campanumoea javanica Blume from Codonopsis plants, and specifically identify Codonopsis sichuanensis (Pai) Hong and Campanumoea javanica Blume. Among them, the Codonopsis sichuanensis (Pai) Hong with characteristic double peaks of 105.2 bp - 105.8 bp and 106.2 bp - 106.8 bp fragments detected; the Campanumoea javanica Blume with characteristic double peaks of 96.2 bp - 96.8 bp and 97.2 bp - 98 bp fragments detected.

[0094] Table 4 shows the capillary electrophoresis results of different origins

[0095]

[0096]

[0097] Finally, the fluorescence PCR system was established and detected. According to the primer screening results, the primers in Table 3 were synthesized with FAM fluorescence labeling at the 5' end. The primers were synthesized by Shanghai Sangon Biotech Co., Ltd. and purified by HPLC.

[0098] Table 5 shows the fluorescence primer information table

[0099]

[0100] The PCR products were sent to Ruibo Biotechnology Co., Ltd. for detection. The allele sizes were determined with reference to the GS-LIZ500 molecular weight internal standard using GeneMarker.

[0101] In summary, the identification method of Codonopsis sichuanensis (Pai) Hong provided by the present invention can be used for the specific identification of Codonopsis sichuanensis (Pai) Hong. It includes extracting genomic DNA, using the above genomic DNA as a template, performing PCR amplification with the designed primers, and detecting the amplified sequences by capillary electrophoresis. The allele sizes are determined with reference to the GS-LIZ500 molecular weight internal standard using GeneMarker. If characteristic double peaks of 105.2 bp - 105.8 bp and 106.2 bp - 106.8 bp fragments are detected, then the species is Codonopsis sichuanensis (Pai) Hong. If characteristic double peaks of 96.2 bp - 96.8 bp and 97.2 bp - 98 bp fragments are detected, then the species is Campanumoea javanica Blume.

[0102] The above are the preferred embodiments of the invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications are also regarded as the protection scope of the present invention.

Claims

1. A primer set, characterized in that, The primer set includes an upstream primer with a sequence as shown in SEQ ID NO: 1 and a downstream primer with a sequence as shown in SEQ ID NO: 2; Among them, a fluorescent labeling group is attached to the 5'-end of the upstream primer, and the fluorescent labeling group is selected from one of fluorescein FAM, fluorescein ROX, fluorescein HEX, and fluorescein TAMARA.

2. The application of the primer set according to claim 1 in (1) or (2): (1) Identifying the authenticity of Codonopsis tangshen Oliv; (2) Specifically identifying Codonopsis tangshen Oliv and Campanumoea javanica Bl. from Codonopsis pilosula (Franch.) Nannf., Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen, Codonopsis tangshen Oliv, and Campanumoea javanica Bl.

3. Use of the primer set according to claim 2, characterized in that, It includes the following steps: Extracting the genomic DNA of the sample to be tested; Using the genomic DNA of the sample to be tested as a template, performing PCR amplification with the upstream primer with a sequence as shown in SEQ ID NO: 1 and the downstream primer with a sequence as shown in SEQ ID NO: 2 to obtain a PCR amplification product; Performing capillary electrophoresis detection on the PCR amplification product; If characteristic double peaks of 105bp - 106bp and 106bp - 107bp fragments appear in the detection, the sample to be tested is Codonopsis tangshen Oliv; If characteristic double peaks of 96bp - 97bp and 97bp - 98bp fragments appear in the detection, the sample to be tested is Campanumoea javanica Bl.

4. A method for identifying Codonopsis tangshen Oliv., characterized in that, It includes the following steps: Extracting the genomic DNA of the sample to be tested; Using the genomic DNA of the sample to be tested as a template, performing PCR amplification with the upstream primer with a sequence as shown in SEQ ID NO: 1 and the downstream primer with a sequence as shown in SEQ ID NO: 2 to obtain a PCR amplification product; Performing capillary electrophoresis detection on the PCR amplification product, and judging whether the sample to be tested is Codonopsis tangshen Oliv according to the detection result.

5. The identification method of Codonopsis tangshen Oliv. medicinal materials according to claim 4, characterized in that The program of the PCR amplification is: pre-denaturing the amplification system at a temperature of 94°C - 96°C for 2 min - 4 min; Denaturing at a temperature of 94°C - 96°C for 5 s - 15 s, then annealing at a temperature of 50°C - 60°C for 5 s - 15 s, and then extending at a temperature of 70°C - 75°C for 5 s - 15 s, repeating the denaturing, annealing, and extending treatments 33 - 36 times each; Extending at a temperature of 70°C - 75°C for 4 min - 6 min and then completing the preservation at 3°C - 5°C.

6. The identification method of Codonopsis tangshen Oliv. medicinal materials according to claim 4, characterized in that, The reaction system for the PCR amplification includes: 9 μL - 11 μL of 2×M5 Supper FastTaq PCR, 0.2 μL - 0.8 μL of the upstream primer, 0.2 μL - 0.8 μL of the downstream primer, 0.5 μL - 1.5 μL of the DNA template, and sterile double-distilled water is added to make up to 20 μL.

7. The identification method of Codonopsis tangshen Oliv. according to claim 6, characterized in that, The concentration of the DNA template is 25 ng / μl - 35 ng / μl; The concentration of the upstream primer is 8 μmol / L - 12 μmol / L; The concentration of the downstream primer is 8 μmol / L - 12 μmol / L.

8. The identification method of Codonopsis tangshen Oliv. as described in claim 4, characterized in that, The electrophoretic detection includes: detecting the amplified sequence by capillary electrophoresis.

9. The identification method of Codonopsis tangshen Oliv. as described in claim 4, characterized in that, If characteristic double peaks of 105 bp - 106 bp and 106 bp - 107 bp fragments appear in the detection result, the sample to be tested is Codonopsis tangshen Oliv. medicinal material; otherwise, the sample to be tested is a fake or inferior product.

10. The identification method of Codonopsis tangshen Oliv. as described in claim 4, characterized in that, If characteristic double peaks of 96 bp - 97 bp and 97 bp - 98 bp fragments appear in the detection result, the sample to be tested is Campanumoea javanica Blume medicinal material.

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