Pinellia ternate SSR molecular marker, primer group and application thereof in construction of pinellia ternate DNA fingerprint spectrum

Abstract

The invention belongs to the technical field of plant germplasm resource identification, and particularly relates to a pinellia ternata SSR molecular marker, a primer group and application of the pinellia ternata SSR molecular marker in construction of a pinellia ternata DNA fingerprint spectrum. Specifically, 17 pairs of SSR primers are utilized to carry out genetic diversity analysis on 255 parts of pinellia ternata germplasm resources, the usage amount of the primers is further reduced by utilizing polymorphic information content, 14 pairs of core SSR primers are screened out, and the DNA fingerprint spectrum of the 255 parts of pinellia ternata germplasm resources is successfully constructed. Theoretical and technical support is provided for identification and classification of pinellia ternate germplasm resources, germplasm innovation and new variety breeding, so that the method has good practical application value.

Description

Technical Field

[0001] The invention belongs to the technical field of plant germplasm resource identification, and specifically relates to a pinellia SSR molecular marker, a primer set and an application thereof in constructing a pinellia DNA fingerprint. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Pinellia ternata (Thunb.) Breit. is a perennial herbaceous plant of the Araceae family. It was first recorded in the Book of Rites, "The pinellia grows in May, which is the middle of summer, hence the name". Pinellia ternata is a famous local medicinal material in Hubei. Its dried tubers are used as medicine. It has a long history of medicinal use and has the effects of drying dampness and resolving phlegm, relieving adverse reactions and stopping vomiting. Pinellia ternata is widely distributed in my country and grows in areas below 2,500 meters above sea level. It is mainly produced in Hubei, Gansu, Shanxi, Henan, Hebei, Sichuan, Guizhou, Anhui, Jiangsu, Shandong, Zhejiang and other regions.

[0004] Research on Pinellia ternata mainly focuses on chemical composition, pharmacological effects, quality standards, clinical and daily applications, and there are few studies on the genetic diversity of Pinellia ternata in molecular biology. In recent years, with urbanization and economic development, the habitat of Pinellia ternata is decreasing. With excessive excavation and abuse of herbicides by humans, wild resources have also decreased sharply. In addition, the sources of Pinellia ternata seedlings used for production and cultivation in different origins are also unclear. Field observations have found that the germplasm resources of Pinellia ternata in the main planting areas and the real production areas are very chaotic. Multiple phenotypes of Pinellia ternata can be found in a small area, resulting in unstable yield and quality. Therefore, high-yield, high-resistance, and high-quality Pinellia ternata varieties are urgently needed to be cultivated. It is very important to develop a set of molecular markers with strong identification ability and good reproducibility to quickly identify and classify Pinellia ternata varieties and protect wild and existing resources.

[0005] Simple Sequence Repeats (SSR), also known as microsatellite DNA, is a type of tandem repeat sequence consisting of several nucleotides (generally 1-6) as repeating units. It is highly favored due to its high polymorphism, good stability, multiple alleles, co-dominance, abundant quantity and simple operation. In recent years, it has been widely used in germplasm resource research, hybrid identification, genetic map construction, target gene positioning, genetic diversity research, etc.

[0006] Compared with morphological identification methods, DNA fingerprinting has a high degree of individual specificity and environmental stability. DNA fingerprinting is one of the technical means that can effectively identify strains. DNA fingerprinting is based on nucleotide site variation within individuals, and identifies and distinguishes strains according to the difference in genome characteristics between different strains. Developing molecular markers is a key link in constructing fingerprints. However, markers have not yet been used to construct a large-scale DNA fingerprint of Pinellia germplasm resources, and the DNA fingerprint of Pinellia germplasm resources plays a very important role in the identification of germplasm resources. However, the inventors found that there are still few reports on the analysis and research of Pinellia germplasm resources based on SSR technology. Summary of the invention

[0007] In view of the deficiencies in the prior art, the present invention provides a Pinellia SSR molecular marker, a primer set and its application in constructing a Pinellia DNA fingerprint. Specifically, the present invention uses 17 pairs of SSR primers to carry out genetic diversity analysis of 255 Pinellia germplasm resources, further simplifies the usage of primers using polymorphic information content, screens out 14 pairs of core SSR primers, constructs a DNA fingerprint of 255 Pinellia germplasm resources, and simultaneously lays a foundation for accurate identification of Pinellia germplasm resources and breeding of superior varieties. Based on the above research results, the present invention is completed.

[0008] Specifically, the present invention relates to the following technical solutions:

[0009] In a first aspect of the present invention, a Pinellia ternata SSR molecular marker is provided, wherein the Pinellia ternata SSR molecular marker comprises the following SSR loci: SSR45, SSR44, SSR95, SSR34, SSR46, SSR72, SSR04, SSR68, SSR14, SSR40, SSR80, SSR02, SSR42, SSR93, SSR104, SSR25, and SSR55;

[0010] The above-mentioned Pinellia SSR molecular markers were obtained by amplification of the following primer pairs: SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO.9-10, SEQ ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO.19-20, SEQ ID NO.21-22, SEQID NO.23-24, SEQ ID NO.25-26, SEQ ID NO.27-28, SEQ ID NO.29-30, SEQ ID NO.31-32 and SEQ ID NO.31-34.

[0011] Furthermore, the Pinellia SSR molecular markers are a group consisting of SSR45, SSR44, SSR95, SSR34, SSR46, SSR72, SSR04, SSR68, SSR14, SSR40, SSR02, SSR93, SSR25 and SSR55. The Pinellia SSR molecular markers are of high quality, strong representativeness, and high genetic variation, and can achieve strain differentiation for 255 Pinellia germplasm resources.

[0012] The second aspect of the present invention provides a Pinellia ternata SSR molecular marker primer set, which comprises any one or more of the following 17 pairs of specific primers, and the nucleotide sequences thereof are shown in SEQ ID NO.1-34 respectively.

[0013] Furthermore, the Pinellia ternata SSR molecular marker primer set comprises the following 14 pairs of specific primer pairs, whose nucleotide sequences are SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO.9-10, SEQ ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO.19-20, SEQ ID NO.23-24, SEQ ID NO.27-28, SEQ ID NO.31-32 and SEQ ID NO.33-34.

[0014] The third aspect of the present invention provides a product for detecting the above-mentioned Pinellia ternata SSR molecular marker, wherein the product at least comprises the above-mentioned Pinellia ternata SSR molecular marker primer set;

[0015] Furthermore, the product may be a detection kit, a gene chip, a detection device, a detection equipment, etc., which are not specifically limited here.

[0016] The fourth aspect of the present invention provides the use of the above-mentioned Pinellia SSR molecular marker, Pinellia SSR molecular marker primer set, and the product for detecting the above-mentioned Pinellia SSR molecular marker in any one or more of the following:

[0017] 1) Genetic diversity analysis of Pinellia ternata;

[0018] 2) Construction of the genetic map of Pinellia ternata;

[0019] 3) Identification of Pinellia ternata germplasm resources;

[0020] 4) Analysis of the relationship of Pinellia ternata;

[0021] 5) Mining of functional genes in Pinellia ternata;

[0022] 6) Molecular marker-assisted breeding of Pinellia ternata.

[0023] In another specific embodiment of the present invention, in said 2), constructing the genetic map of Pinellia ternata is specifically constructing a DNA fingerprint map of Pinellia ternata.

[0024] In another specific embodiment of the present invention, in said 3), the identification of Pinellia ternata germplasm at least includes the identification of Pinellia ternata varieties.

[0025] The fifth aspect of the present invention provides a method for constructing a DNA fingerprint of Pinellia ternata, the steps comprising: extracting the genomic DNA of the Pinellia ternata sample to be tested, performing PCR amplification on the genomic DNA of the Pinellia ternata to be tested using the above-mentioned Pinellia ternata SSR molecular marker primer set, performing capillary electrophoresis detection on the PCR products, and constructing a DNA fingerprint of Pinellia ternata.

[0026] Furthermore, the Pinellia sample to be tested may specifically be young and tender leaves of Pinellia.

[0027] In the construction method, when the SSR molecular marker primer set performs PCR amplification on the genomic DNA of the Pinellia ternata to be tested, the forward primer in each primer pair is connected to a universal M13 adapter sequence;

[0028] The PCR amplification procedure includes: 94°C pre-denaturation for 5 min, 94°C denaturation for 30 s, 55°C annealing for 30 s, 72°C extension for 30 s, a total of 35 cycles; and a final 72°C end extension for 5 min and storage at 4°C.

[0029] Furthermore, the PCR amplification is fluorescent PCR amplification, which can be specifically achieved by adding a fluorescent modification group into the PCR amplification system. The fluorescent modification group includes any one or more of ROX, HEX, FAM and TAMRA, which are not specifically limited here.

[0030] Furthermore, the construction method also includes statistical analysis of the capillary electrophoresis detection results, so as to perform genetic diversity analysis of Pinellia ternata, cluster analysis polymorphic information content (PIC) calculation and construction of DNA fingerprint map.

[0031] Furthermore, the method for constructing the DNA fingerprint of Pinellia ternata includes: converting all alleles into 1 / 0 format, with 1 and 0 representing the corresponding fragments detected and undetected respectively. In a specific embodiment of the present invention, the Pinellia ternata fingerprint includes 255 Pinellia ternata varieties.

[0032] Beneficial technical effects of one or more of the above technical solutions:

[0033] (1) The SSR molecular markers developed by the above technical scheme through the transcriptome data of Pinellia ternata have high polymorphism potential and are highly practical in germplasm identification. Through the experiment, 17 pairs of SSR primers were screened, and the genetic diversity of 255 Pinellia ternata germplasm resources were analyzed using SSR technology, laying the foundation for the construction of Pinellia ternata core germplasm and the breeding and utilization of germplasm resources.

[0034] (2) The above technical scheme uses SSR molecular markers and the germplasms collected in the Pinellia germplasm resource nursery as test materials. Through capillary electrophoresis detection, the usage of primers is further simplified according to the polymorphic information content of the primers, and 14 pairs of core markers are screened out. The first DNA fingerprint map containing 255 Pinellia germplasms was established. This method is fast, accurate, precise, reproducible and stable, and can identify multiple varieties at the same time. It is convenient and fast. At the same time, it provides theoretical and technical support for the identification and classification of Pinellia germplasm resources, germplasm innovation and new variety breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 This is the peak diagram of capillary fluorescence electrophoresis detection of partial amplification using primer SSR72 in the present invention.

[0036] Figure 2 This is a cluster analysis result diagram of the Pinellia ternata germplasm of the present invention. Germplasm resources are marked with two background colors: sky blue represents wild germplasm resources, and flesh color represents cultivated germplasm resources. DETAILED DESCRIPTION

[0037] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0038] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0039] The present invention is further described in conjunction with specific examples. The following examples are only for explaining the present invention and are not intended to limit the content thereof. If the specific experimental conditions are not specified in the examples, they are usually carried out under conventional conditions or under conditions recommended by the sales company; the materials and reagents used in the examples, unless otherwise specified, can be purchased through commercial channels.

[0040] The present invention is further explained by the following examples, but they are not intended to limit the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0041] Table 1: Pinellia ternata germplasm resources

[0042]

[0043]

[0044]

[0045]

[0046] 2.SSR Primer Development

[0047] A. Using the existing Pinellia ternata transcriptome sequence, the MISA script (http: / / pgrc.ipk-gatersleben.de / misa / ) was used to scan and screen SSR sites to obtain a large number of SSR sites; and Primer 3 software was used to design primers for the sites.

[0048] B. In the present invention, the repetitive motif of SSR is analyzed according to the following standards: the number of repeated dinucleotides ≥ 6; the number of repeated trinucleotides, tetranucleotides, pentanucleotides and hexanucleotides ≥ 5. Specific SSR primers were designed using Prime 3, and the parameters were set as follows: primer length 18-27bp; annealing temperature 55-65°C; PCR product length range between 100bp and 400bp; GC content between 40% and 60%, and finally 119 pairs of SSR primers were successfully designed. The amplification efficiency and polymorphism of primers were studied using 8 samples, and finally 17 pairs of primers that can stably amplify clear and polymorphic bands were screened out from 119 pairs of primers. The primer sequences are shown in Table 2 and were commissioned to be synthesized by Sangon Biotech (Shanghai) Co., Ltd.

[0049] 3. Genomic DNA Extraction

[0050] 255 healthy young leaves of materials were selected, and their DNA was extracted using the improved CTAB method. The DNA concentration and purity of the test materials were detected using Nanodrop nucleic acid analyzer and 1% agarose gel electrophoresis. The qualified DNA samples were stored at -20℃ for later use.

[0051] 4. Capillary electrophoresis detection

[0052] A. Add the linker sequence M13"GTAAAA to the 5' end of the forward primer of 17 pairs of SSR primers.

[0053] CGACGGCCAGT", and purified using PAGE to obtain the M13 linker forward primer; modified M13 linkers with four different fluorescent labels (FAM / HEX / ROX / TAMRA) were synthesized.

[0054] B. Using the genomic DNA of the sample to be tested extracted in step 3 as a template, PCR amplification was performed with a reverse primer and an M13 adapter forward primer to obtain a fluorescent PCR amplification product: the PCR amplification system is as follows: 2 μl of a DNA template of 50 ng / μl, 5 μl of 2×Es Taq MsaterMix, 0.04 μl of an M13 adapter forward primer at a concentration of 10 pmol / μl, 0.25 μl of a reverse primer at a concentration of 10 pmol / μl, 0.15 μl of an M13 fluorescent modification group (ROX or HEX or FAM or TAMRA), and the rest was filled to 10 μl with sterile water. The reaction procedure for PCR amplification is: 94°C pre-denaturation for 5 min, 94°C denaturation for 30 s, 55°C annealing for 30 s, 72°C extension for 30 s, a total of 35 cycles; final 72°C end extension for 5 min, and storage at 4°C.

[0055] C. The fluorescent PCR amplification product obtained in step B was detected by capillary fluorescence electrophoresis using a DNA sequencer ABI3730XL, and the sample fragment size and the number of alleles were read using GeneMapper 4.0 to count the band detection results.

[0056] 5. Data collation

[0057] The size of the amplified fragments obtained by comprehensive sorting was used to calculate the following genetic diversity parameters of each primer pair, such as the number of effective alleles (Ne), Shannon information index (I) and Nei's genetic diversity index (H). After calculating the alleles of different primers with Popgen32 software, the polymorphic information content (PIC) of the SSR loci was calculated with PIC_CALC software. According to the size of the amplified fragments obtained by comprehensive sorting, the amplified fragments were numbered and read from large to small, and the presence of amplified bands was recorded as "1" and the absence of bands was recorded as "0". The original matrix was established and imported into MEGA11 software to draw the NJ tree, and cluster analysis was performed on the 255 Pinellia samples. The markers were further simplified according to the polymorphic information content (PIC) of the markers to construct the standard fingerprint map of each variety.

[0058] Example 1: Design of SSR primers

[0059] According to the above-mentioned SSR primer development method, SSR markers were screened, and a total of 119 pairs were screened; using capillary electrophoresis detection, 17 pairs of primers with high polymorphism were screened, as shown in Table 2, corresponding to sequences SEQ NO ID.1~34.

[0060] Table 2: Information of 17 pairs of SSR primers

[0061]

[0062]

[0063] Capillary electrophoresis was used to detect the amplification of 17 pairs of SSR primers in 255 germplasm resources. Some of the results are shown in Table 3. A total of 125 polymorphic bands were amplified, with 3 to 13 alleles for each primer pair, and the gene fragments were between 150 and 400 bp; the effective allele number (Ne) ranged from 1.1183 to 1.7295, with an average of 1.3864 effective alleles; Nei's genetic diversity index (H) ranged from 0.1012 to 0.4114, with an average of 0.2373; Shannon information diversity index (I) ranged from 0.1981 to 0.5988, with an average of 0.3719; and polymorphic information content (PIC) ranged from 0.3516 to 0.8390, with an average of 0.6482. These primers showed good polymorphism and were suitable for genetic diversity analysis and germplasm resource identification in Pinellia ternata.

[0064] Table 3: Statistics of amplification information of 17 pairs of SSR primers in 255 Pinellia ternata materials

[0065]

[0066]

[0067] According to the cluster dendrogram ( Figure 2), 255 Pinellia accessions were divided into two clusters (Cluster 1 and Cluster 2). Cluster 1 contained 157 accessions, including 44 cultivated accessions from Qianjiang, 3 cultivated accessions from Tianmen, and 27 wild accessions from Hubei (Shiyan (6), Enshi (4), Yichang (14), Huangshi (1), Huanggang (1), and Suizhou (1)). The other 83 accessions came from other provinces, including Hunan (26), Yunnan (9), Sichuan (3), Henan (6), Zhejiang (6), Gansu (3), Jiangxi (5), Hebei (4), Henan (6), Zhejiang (6), Gansu (3), Jiangxi (5), Hebei (4), Shandong (2), Guizhou (5), Shaanxi (1), Beijing (3), Anhui (3), Jiangsu (3), Fujian (2), Chongqing (1), and Shanxi (1). Group 2 contained 98 accessions, of which 78 were from Hubei, including 71 cultivated accessions from Qianjiang, 4 cultivated accessions from Tianmen, and 3 wild accessions from other areas of Hubei (Enshi (2), Shennongjia (1). The others were from Hunan (2), Yunnan (2), Sichuan (5), Zhejiang (1), Gansu (3), Shandong (3), Jiangxi (2), and Shaanxi (2). Most of the wild accessions of Pinellia ternata from different origins were clustered into Group 1, and wild accessions from different origins tended to cluster into one category. For example, some wild accessions from Hunan were clustered into one category, and wild accessions from Beijing, Yichang, Enshi, Sichuan, Zhejiang, and Jiangsu were clustered into one category respectively. However, The nine germplasms from Yunnan were more dispersed. The proportion of cultivated germplasm in cluster 2 was higher, specifically 79.59%. In addition, wild germplasm from Sichuan and Shanxi also appeared in this cluster, and each clustered into a branch. The results of cluster analysis showed that most germplasm resources were clustered according to whether the resources were wild or cultivated. Wild resources tended to be concentrated in cluster 1. However, the dendrogram showed a more continuous pattern without obvious branches. This result suggests that there is already a genetic distance between cultivated and wild germplasm, but there is no obvious difference. This may be due to various human interventions or other abiotic factors that cause genotypes to migrate from one place to another.

[0068] Example 2: Fingerprints and identification methods of 255 Pinellia ternata germplasm resources

[0069] According to the principle of the minimum markers for distinguishing 255 germplasms and the polymorphic information content value of the markers, 14 pairs of core markers (SSR45, SSR44, SSR95, SSR34, SSR46, SSR72, SSR04, SSR68, SSR14, SSR40, SSR02, SSR93, SSR25 and SSR55) were further screened from the 17 pairs of SSR markers to construct a molecular fingerprint library of the 255 germplasms. Statistics were performed based on whether there were obvious bands at the migration position of the amplified bands, where 1 indicated the presence of electrophoresis bands and 0 indicated the absence of electrophoresis bands, i.e., all alleles were converted into 1 / 0 format. The DNA fingerprints of the 255 Pinellia germplasms are shown in Table 4.

[0070] Table 4: DNA fingerprints of 255 Pinellia accessions

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081] The method of using the above fingerprint map to distinguish the strains of Pinellia germplasm resources is to count whether there are obvious bands at the migration position of SSR45, SSR44, SSR95, SSR34, SSR46, SSR72, SSR04, SSR68, SSR14, SSR40, SSR02, SSR93, SSR25, and SSR55 amplified bands, where 1 indicates the presence of an electrophoresis band and 0 indicates the absence of an electrophoresis band, that is, all alleles are converted into a 1 / 0 format, with 1 and 0 representing the corresponding fragments detected and not detected, respectively, to obtain a 113-bit 1 / 0 format string, which can be compared with the fingerprint map shown in Table 4.

[0082] The above fingerprint map can be used to distinguish the strains of 255 Pinellia germplasm resources. Each variety has a unique and definite fingerprint map with high accuracy and specificity.

[0083] It is worth noting that the above 14 pairs of core markers have established the first DNA fingerprint information database containing 255 Pinellia germplasms, providing an effective way to accurately identify Pinellia germplasm resources; and any removal of any pair of the 14 pairs of core markers will cause the fingerprint map to become invalid, and it will be impossible to accurately identify Pinellia germplasm resources. Adding any pair of the 14 pairs of core markers will not have a beneficial effect on the identification.

[0084] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A Pinellia ternata SSR molecular marker, characterized in that: The pinellia SSR molecular markers include the following SSR loci: SSR45, SSR44, SSR95, SSR34, SSR46, SSR72, SSR04, SSR68, SSR14, SSR40, SSR80, SSR02, SSR42, SSR93, SSR104, SSR25, and SSR55; The Pinellia ternata SSR molecular markers were obtained by amplifying the following primer pairs: SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO.9-10, SEQ ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO.19-20, SEQ ID NO.21-22, SEQ ID NO.23-24, SEQ ID NO.25-26, SEQ ID NO.27-28, SEQ ID NO.29-30, SEQ ID NO.31-32 and SEQ ID NO.31-34.

2. The Pinellia ternata SSR molecular marker according to claim 1, characterized in that: The Pinellia SSR molecular markers are a group consisting of SSR45, SSR44, SSR95, SSR34, SSR46, SSR72, SSR04, SSR68, SSR14, SSR40, SSR02, SSR93, SSR25 and SSR55.

3. A Pinellia ternata SSR molecular marker primer set, characterized in that: The Pinellia ternata SSR molecular marker primer set comprises any one or more of the following 17 pairs of specific primer pairs, and the nucleotide sequences of the specific primer pairs are shown in SEQ ID NOs. 1-34 respectively; Furthermore, the Pinellia ternata SSR molecular marker primer set comprises the following 14 pairs of specific primer pairs, whose nucleotide sequences are SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO.9-10, SEQ ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO.19-20, SEQID NO.23-24, SEQ ID NO.27-28, SEQ ID NO.31-32 and SEQ ID NO.33-34.

4. A product for detecting the SSR molecular marker of Pinellia ternata according to claim 1 or 2, characterized in that: The product at least contains the Pinellia ternata SSR molecular marker primer described in claim 3.

5. The product according to claim 4, characterized in that The product is a detection kit, a gene chip, a detection device or a detection equipment.

6. Use of the Pinellia ternata SSR molecular marker according to claim 1 or 2, the Pinellia ternata SSR molecular marker primer set according to claim 3, and the product according to claim 4 in any one or more of the following: 1) Genetic diversity analysis of Pinellia ternata; 2) Construction of the genetic map of Pinellia ternata; 3) Identification of Pinellia ternata germplasm resources; 4) Analysis of the relationship of Pinellia ternata; 5) Mining of functional genes in Pinellia ternata; 6) Molecular marker-assisted breeding of Pinellia ternata.

7. The use according to claim 6, characterized in that In the above 2), constructing the genetic map of Pinellia ternata specifically comprises constructing a DNA fingerprint of Pinellia ternata; In the above 3), the identification of Pinellia ternata germplasm at least includes the identification of Pinellia ternata varieties.

8. A method for constructing a DNA fingerprint of Pinellia ternata, characterized in that: The steps include: Extract the genomic DNA of the Pinellia sample to be tested, perform PCR amplification on the genomic DNA of the Pinellia sample to be tested using the Pinellia SSR molecular marker primer set described in claim 3, perform capillary electrophoresis detection on the PCR product, and construct a Pinellia DNA fingerprint.

9. The construction method according to claim 8, characterized in that: The pinellia samples to be tested are young and tender leaves of pinellia.

10. The construction method according to claim 9, characterized in that: In the construction method, when the SSR molecular marker primer set performs PCR amplification on the genomic DNA of the Pinellia ternata to be tested, the forward primer in each primer pair is connected to a universal M13 adapter sequence; Furthermore, the PCR amplification is fluorescent PCR amplification, specifically adding a fluorescent modification group to the PCR amplification system; Furthermore, the construction method also includes statistical analysis of the capillary electrophoresis detection results, so as to perform genetic diversity analysis of Pinellia ternata, cluster analysis polymorphic information content PIC calculation and DNA fingerprint map construction; Furthermore, the method for constructing the DNA fingerprint of Pinellia ternata includes: converting all alleles into a 1 / 0 format, with 1 and 0 representing the corresponding fragments that are detected and not detected, respectively.

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