KR101974175B1 - SNP Marker derived from complete sequencing of chloroplast genome of seven Panax species, primer set for discrimination of Panax species and uses thereof - Google Patents

Abstract

The present invention relates to a primer set and an SNP marker for identification of species based on complete sequencing of chloroplast genome sequences of seven species of Panax sp., and uses thereof. The present invention enables highly reliable verification of Panax species and enables identification thereof at a lower cost and higher efficiency than conventional methods. It is also possible to maintain purity of Panax species, protect varieties, and solve conflicts related to seeds.

Description

SNP markers and primer sets for distinguishing chloroplast genomes from seven kinds of genomes in ginseng genus and their uses {SNP Marker derived from complete sequencing of chloroplast genome of seven Panax species, primer set for discrimination of Panax species and uses thereof

The present invention relates to a complete detoxification-based interspecific SNP marker and primer set of seven kinds of chloroplast genomes of ginseng genus and its use.

In addition to Korean ginseng, American ginseng, Chinese ginseng, Chinese ginseng, Vietnamese ginseng, ginseng ginseng, and lesional ginseng have been widely used for thousands of years as medicinal crops in Asia and North America. Because of the high value of pharmacological action and expensive value, ginseng plants are often mixed with other plants in ginseng or they distribute ginseng, but it is difficult to distinguish them because various morphological characteristics may occur depending on the environment in which the plant grows . In addition, since they are distributed in the form of root slices, powder, or product, existing methods have limitations in discriminating whether or not each species is stompered, modulated, or mixed. Therefore, in order to discriminate them more precisely, economical, highly accurate, and convenient methods are required, and molecular markers based on DNA sequence are considered suitable.

Previously used nuclear genome-based DNA molecular markers have been used to analyze intraspecific diversity rather than interspecific diversity due to the inherent limitations of nuclear variants. On the other hand, the chloroplast genome is considered to be more suitable for species discrimination because there are relatively few varieties in the species and a large number of copies in the cytoplasm. Plant DNA bar coding markers in the upstream region of the chloroplast that were previously used frequently have mutations that can occur in the species due to frequent occurrence in the species. Therefore, it is necessary to develop new markers without mutation in the species .

Therefore, in the present invention, it was aimed to search SNP regions having high polymorphism between plants by comparing complete chloroplast sequences of seven species of ginseng genus, and to develop a molecular marker in this region to effectively distinguish each species. In addition to the conventional method of discriminating based on agarose gel, a molecular marker using a fluorescent label (Kompetitive Allele Specific PCR) was also developed so that time and labor for analysis can be reduced and applied to a mass analysis system.

On the other hand, Korean Patent No. 1426466 discloses " Koryo ginseng chloroplast genome based complete variety-based varieties, species discrimination markers and primer sets and kits containing them ", Korean Patent Publication No. 2016-0057258 discloses' A complete detoxification-based species discrimination marker and a primer set and a use thereof of a genome and a complete rDNA sequence in the nucleus have been disclosed, Set " and " use thereof " have not been described.

The present invention is derived by the request as described above, the present inventors have found that ginseng (Panax) 7 plant species (Korean ginseng (Panax ginseng), American ginseng (P. quinquifolius), jeonchilsam (P. notoginseng), jukjeol three ( P. japonicus , P. vietnamensis , P. trifolius , and Panax stipuleanatus ) were completely decoded, and based on mutation regions discovered through comparative genomic analysis, We identified 19 SNPs that can distinguish each species. In addition, using the dCAPS primer set and the KASP primer set composed of 18 primer sets capable of analyzing 18 SNPs among the 19 SNPs developed in the present invention, the ginseng saplings such as Korean ginseng, American Sampler, The present invention has been accomplished by confirming the fact that seven kinds of ginseng species can be identified at low cost and high efficiency than those of the conventional methods.

In order to solve the above-described problems, the present invention provides a method for detecting SNP (single nucleotide polymorphism) located at the 249th base in the nucleotide sequence of rpl20 ( ribosomal protein L20 ) gene of SEQ ID NO: 1 of Panax ginseng. A SNP located at the 69th base, the 187th base, or the 213 base of the nucleotide sequence of the ndhK ( NADH - plastoquinone oxidoreductase subunit K ) gene of SEQ ID NO: 2; A SNP located at the 56th base of the nucleotide sequence of the rps15 ( ribosomal protein S15 ) gene of SEQ ID NO: 3; A SNP located at the 310th or 831st base in the nucleotide sequence of the ndhA ( NADH - plastoquinone oxidoreductase subunit 1 ) gene of SEQ ID NO: 4; SNPs located at the 996th , 1044th , 1362th , 1658th , or 1815th bases of the nucleotide sequence of the rpoC1 ( RNA polymerase beta subunit ) of SEQ ID NO: 5; Of the base sequence of rpoC2 (RNA polymerase beta subunit) gene of SEQ ID NO: 6 in the 1263rd, 1320th or 2145th base SNP; A SNP located at the 1158th or 3186th base in the nucleotide sequence of the rpoB ( RNA polymerase beta subunit ) gene of SEQ ID NO: 7; The ndhH of SEQ ID NO: 8 ( NADH - plastoquinone oxidoreductase subunit 7 ) SNP located in nucleotide 171 of the nucleotide sequence; Or a SNP located in the 414 base sequence of the nucleotide sequence of psbB ( photosystem II CP47 chlorophyll apoprotein ) gene of SEQ ID NO: 9, or a complementary polynucleotide of the polynucleotide consisting of 8 or more consecutive nucleotides thereof. Panax ginseng , P. quinquifolius , P. notoginseng , P. japonicus , P. vietnamensis , P. trifolius and Panax stipuleanatus , The present invention provides a single nucleotide polymorphism (SNP) marker composition for distinguishing one or more species from a genus of a genus including ginseng.

The present invention also provides a kit comprising at least one primer set selected from the group consisting of SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, and oligonucleotide primer sets of SEQ ID NOs: 14 and 15; At least one primer set selected from the group consisting of SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, and oligonucleotide primer sets of SEQ ID NOs: 20 and 21; At least one primer set selected from the group consisting of SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, and oligonucleotide primer sets of SEQ ID NOs: 26 and 27; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NOS: 28 and 29 and SEQ ID NOS: 30 and 31; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NOS: 32 and 33 and SEQ ID NOS: 34 and 35; At least one primer set selected from the group consisting of SEQ ID NOs: 36 and 37, SEQ ID NOs: 38 and 39, and oligonucleotide primer sets of SEQ ID NOs: 40 and 41; And an oligonucleotide primer set of SEQ ID NOs: 42 and 43 and SEQ ID NOs: 44 and 45. The primer set of Korean Ginseng, American Ginseng, American Chrysanthemum, Chrysanthemum morifolium, A set of derived cleaved amplified polymorphic sequence (dCAPS) primers for distinguishing one or more species from species of ginseng.

Also, the present invention provides a kit comprising at least one primer set selected from the group consisting of SEQ ID NOs: 46, 47 and 48, SEQ ID NOs: 49, 50 and 51, oligonucleotide primer sets of SEQ ID NOs: 52, 53 and 54; At least one primer set selected from the group consisting of SEQ ID NOs: 55, 56 and 57, SEQ ID NOs: 58, 59 and 60, and oligonucleotide primer sets of SEQ ID NOs: 61, 62 and 63; At least one primer set selected from the group consisting of SEQ ID NOs: 64, 65 and 66, SEQ ID NOs: 67, 68 and 69 and oligonucleotide primer sets of SEQ ID NOs: 70, 71 and 72; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NOs: 73, 74 and 75 and SEQ ID NOs: 76, 77 and 78; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NO: 79, 80 and 81 and SEQ ID NO: 82, 83 and 84; At least one primer set selected from the group consisting of SEQ ID NOs: 85, 86 and 87, SEQ ID NOs: 88, 89 and 90, and oligonucleotide primer sets of SEQ ID NOs: 91, 92 and 93; And a set of oligonucleotide primers of SEQ ID NOS: 94, 95, and 96 and SEQ ID NOS: 97, 98, and 99. The primers set forth in SEQ ID NO: A set of KASP (Kompetitive Allele Specific PCR) primers is provided for distinguishing one or more species from a species of the genus Ginseng containing three foliar ginseng and three lesions of lesion.

The present invention also relates to a method for distinguishing one or more species from a species of ginseng including Korean ginseng, American ginseng, Korean white ginseng, white ginseng, Vietnamese ginseng, ginseng ginseng and lesion ginseng using the dCAPS primer set or KASP primer set ≪ / RTI >

The present invention also relates to the aforementioned dCAPS primer set or KASP primer set; And a reagent for carrying out an amplification reaction, wherein the kit comprises at least one species selected from the group consisting of Korean ginseng, American Samoan, Chinese medicinal herb, Korean white ginseng, Vietnamese ginseng, Korean ginseng and lesional ginseng .

18 molecular markers based on the SNP of the chloroplast genome sequence developed in the present invention are useful for distinguishing seven species of ginseng, Ginseng, Korean Ginseng, American Samoa, Jeollabyte, Janseum, Vietnamese Samoa, It will be possible. Since there are two or more SNP-based species-specific markers in each species, it is possible to clearly distinguish seven species of ginseng from the markers presented. Each marker has been developed in the presence of a large number of intracellular chloroplast genomic sequences to confirm stable amplification and a more accurate marker can be completed since the marker is constructed except for some sequences that are presumed to have been transferred from the chloroplastic genome to the mitochondrial genome. In addition, since it is a dCAPS primer based on interspecies SNPs, it is possible to amplify all of the corresponding sequences and confirm the band length difference on the agarose gel through restriction enzyme treatment. Alternatively, a molecular marker using a fluorescent label ), And it is possible to discriminate correctly compared to the conventional chloroplast marker results which have been used in the past.

Therefore, the present invention enables highly reliable verification of ginseng species, and can identify ginseng species at a lower cost and higher efficiency than the conventional methods. It can also be used to maintain the purity of ginseng species, to protect the variety, and to solve seed conflicts.

Figure 1 shows a complete chloroplast genome of seven ginseng species. Colored boxes represent conserved chloroplast genes classified based on the function of the gene product. The genes located inside the circle are genes that are transcribed in a clockwise direction and the genes located outside the circle represent genes that are transcribed in a counterclockwise direction. The genes belonging to different functional groups are shown in color, and the dotted area of the inner circle indicates the GC content of the chloroplast genome.
Figure 2 shows SNP sites in 79 protein coding genes from seven ginseng species. The inner tracks are 79 chloroplast CDS genes. Track A represents the total SNP of a total of seven ginseng species. BG track is ginseng (Panax ginseng) and American ginseng (P. quinquifolius), jeonchilsam (P. notoginseng), jukjeol three (P. japonicus), Vietnam, three (P. vietnamensis), lesions thirty-seven (Panax stipuleanatus) and compared SNPs in P. trifolius are shown. The red, green, blue, and black lines of each track represent four SNPs (T, A, C, and G nucleotides), and yellow indicates InDel.
Fig. 3 shows a schematic diagram of gene transfer between chloroplast and mitochondrial genome from seven ginseng species. Each gray line of the circle represents the chloroplast genome region, which was inserted into the mitochondrial genome of the indicated region. The color box represents the conserved chloroplast genome and is classified according to the function of the product. The genes located inside the circle are genes that are transcribed in a clockwise direction and the genes located outside the circle represent genes that are transcribed in a counterclockwise direction.
Fig. 4 shows the results of analysis of 18 dCAPS markers derived from the CDS SNP region of the chloroplast genome of seven ginseng species. The 18 dCAPS markers are as follows. : Pgdm1-3, Pqdm4-6, Pndm7-9, Pjdm10 and 11, Pvdm12 and 13, Psdm14-16 and Ptdm17 and 18 are Panax ginseng , P. quinquifolius , P. notoginseng ), P. japonicus , P. vietnamensis , Panax stipuleanatus and P. trifolius , respectively; Pg, Panax ginseng ; Pq, US quinquifolius ; Pn, P. notoginseng ; Pj, P. japonicus ; Pv, Viet Nam ( P. vietnamensis ); Ps, Panax stipuleanatus ; Pt, Trifolius ( P. trifolius ). M, 100-bp DNA leather.
Figures 5a and b show the endpoint fluorescence scatter plot of a Kompetitive Allele Specific PCR (KASP) assay. Seven ginseng species were analyzed by 18 KASP markers. Allele-specific primer 1 was identified as FAM (blue), allele-specific primer 2 as HEX (green), and black data points identified as template-free control. FAM, X-axis fluorescence at 523 nm to 568 nm; HEX, Y axis-fluorescence at 483 nm to 533 nm.

In order to accomplish the object of the present invention, the present invention provides a polynucleotide encoding a single nucleotide polymorphism (SNP) located at the 249th base in the nucleotide sequence of the rpl20 ( ribosomal protein L20 ) gene of SEQ ID NO: 1 of Panax ginseng. A SNP located at the 69th base, the 187th or the 213th base among the nucleotide sequences of the ndhK ( NADH-plastoquinone oxidoreductase subunit K ) gene of SEQ ID NO: 2; A SNP located at the 56th base of the nucleotide sequence of the rps15 ( ribosomal protein S15 ) gene of SEQ ID NO: 3; A SNP located at the 310th or 831st base in the nucleotide sequence of the ndhA ( NADH-plastoquinone oxidoreductase subunit 1 ) gene of SEQ ID NO: 4; SNPs located at the 996th , 1044th , 1362th , 1658th , or 1815th bases of the nucleotide sequence of the rpoC1 ( RNA polymerase beta subunit ) of SEQ ID NO: 5; A SNP located at the 1263rd , 1320th or 2145th base in the nucleotide sequence of rpoC2 ( RNA polymerase beta su Primer_AlleleFAM bunit ) of SEQ ID NO: 6; A SNP located at the 1158th or 3186th base in the nucleotide sequence of the rpoB ( RNA polymerase beta subunit ) gene of SEQ ID NO: 7; A SNP located at nucleotide 171 in the nucleotide sequence of ndhH ( NADH-plastoquinone oxidoreductase subunit 7 ) of SEQ ID NO: 8; Or a SNP located in the 414 base sequence of the nucleotide sequence of psbB ( photosystem II CP47 chlorophyll apoprotein ) gene of SEQ ID NO: 9, or a complementary polynucleotide of the polynucleotide consisting of 8 or more consecutive nucleotides thereof. Panax ginseng , P. quinquifolius , P. notoginseng , P. japonicus , P. vietnamensis , P. trifolius and Panax stipuleanatus , The present invention provides a single nucleotide polymorphism (SNP) marker composition for distinguishing one or more species from a genus of a genus including ginseng.

In one embodiment of the invention, the contiguous nucleotides may be 8 to 100 contiguous nucleotides, but are not limited thereto

As used herein, the term " nucleotide " is a deoxyribonucleotide or ribonucleotide present in single-stranded or double-stranded form and includes analogs of natural nucleotides unless specifically stated otherwise.

The SNP of the present invention is a very stable gene marker that directly affects the species discrimination for distinguishing one or more species from seven ginseng species.

The 19 SNP markers of the present invention can be used for species identification of seven ginseng species. The 249th base, which is the SNP mutation position in the nucleotide sequence of the rpl20 gene of SEQ ID NO: 1 in ginseng; The 69th nucleotide, the 187th nucleotide or the 213th nucleotide, which is the SNP mutation position in the nucleotide sequence of the ndhK gene of SEQ ID NO: 2; The 56th nucleotide at the SNP mutation position in the nucleotide sequence of the rps15 gene of SEQ ID NO: 3; A nucleotide sequence at position 310 or 831 in the nucleotide sequence of the ndhA gene of SEQ ID NO: 4; The 996th , 1044th , 1362th , 1658th or 1815th nucleotide of the nucleotide sequence of the rpoC1 gene of SEQ ID NO: 5, which is the SNP mutation site; The 1263rd , 1320th , or 2145th nucleotide in the nucleotide sequence of the rpoC2 gene of SEQ ID NO: 6; A 1158-th or 3186-th base which is a SNP mutation site in the base sequence of the rpoB gene of SEQ ID NO: 7; The 171st nucleotide at the SNP mutation position in the nucleotide sequence of the ndhH gene of SEQ ID NO: 8; Or the 414th nucleotide position of the SNP mutation position in the nucleotide sequence of the psbB gene of SEQ ID NO: 9.

Using the above-mentioned 19 SNPs, it is possible to distinguish seven kinds of ginseng plants from Korea Ginseng, American Sampler, Jeollabuk-do, Jamsil, Gt; dCAPS < / RTI > or KASP primer sets (see Examples 3 and 4).

The present invention relates to a base mutant at each SNP position in the sequence of SEQ ID NOS: 1 to 9, but when such SNP base mutation is found in double stranded gDNA (genomic DNA), the polynucleotide sequence complementary to the nucleotide sequence And the like. Thus, the base at the SNP position in the complementary polynucleotide sequence is a complementary base. In this regard, all sequences provided herein are based on sequences in the sense strand of the genomic DNA unless otherwise noted.

The present invention also provides a kit comprising at least one primer set selected from the group consisting of SEQ ID NOs: 10 and 11, SEQ ID NOs: 12 and 13, and oligonucleotide primer sets of SEQ ID NOs: 14 and 15; At least one primer set selected from the group consisting of SEQ ID NOs: 16 and 17, SEQ ID NOs: 18 and 19, and oligonucleotide primer sets of SEQ ID NOs: 20 and 21; At least one primer set selected from the group consisting of SEQ ID NOs: 22 and 23, SEQ ID NOs: 24 and 25, and oligonucleotide primer sets of SEQ ID NOs: 26 and 27; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NOS: 28 and 29 and SEQ ID NOS: 30 and 31; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NOS: 32 and 33 and SEQ ID NOS: 34 and 35; At least one primer set selected from the group consisting of SEQ ID NOs: 36 and 37, SEQ ID NOs: 38 and 39, and oligonucleotide primer sets of SEQ ID NOs: 40 and 41; And an oligonucleotide primer set of SEQ ID NOs: 42 and 43 and SEQ ID NOs: 44 and 45. The primer set of Korean Ginseng, American Ginseng, American Chrysanthemum, Chrysanthemum morifolium, A set of derived cleaved amplified polymorphic sequence (dCAPS) primers for distinguishing one or more species from species of ginseng.

The primers of the dCAPS (derived cleaved amplified polymorhic sequence) primer set of the present invention were designed so that the sequence of the amplification product according to SNPs of each species has a restriction enzyme site or not. For example, in the case of SEQ ID NOS: 10 and 11 of the present invention, Korean ginseng has nucleotide A at the corresponding position and G at the other position. According to this single nucleotide polymorphism, the amplification product sequence of Korean ginseng by the corresponding primer has a restriction enzyme site which can be cleaved by Cla I, but the amplification product of the remaining species does not have a restriction enzyme site. Therefore, when the restriction enzyme is treated in the PCR amplification product using the corresponding primer, the amplification product of the ginseng is cut by the restriction enzyme and the amplification product of the remaining species is not truncated, and the product of each species is confirmed by electrophoresis The base types of single nucleotide polymorphisms in each species can be analyzed. The nucleotide sequences of other dCAPS primer sets and restriction enzymes for cleavage of the PCR products are as shown in Table 4.

Also, the present invention provides a kit comprising at least one primer set selected from the group consisting of SEQ ID NOs: 46, 47 and 48, SEQ ID NOs: 49, 50 and 51, oligonucleotide primer sets of SEQ ID NOs: 52, 53 and 54; At least one primer set selected from the group consisting of SEQ ID NOs: 55, 56 and 57, SEQ ID NOs: 58, 59 and 60, and oligonucleotide primer sets of SEQ ID NOs: 61, 62 and 63; At least one primer set selected from the group consisting of SEQ ID NOs: 64, 65 and 66, SEQ ID NOs: 67, 68 and 69 and oligonucleotide primer sets of SEQ ID NOs: 70, 71 and 72; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NOs: 73, 74 and 75 and SEQ ID NOs: 76, 77 and 78; At least one primer set selected from the group consisting of oligonucleotide primer sets of SEQ ID NO: 79, 80 and 81 and SEQ ID NO: 82, 83 and 84; At least one primer set selected from the group consisting of SEQ ID NOs: 85, 86 and 87, SEQ ID NOs: 88, 89 and 90, and oligonucleotide primer sets of SEQ ID NOs: 91, 92 and 93; And a set of oligonucleotide primers of SEQ ID NOS: 94, 95, and 96 and SEQ ID NOS: 97, 98, and 99. The primers set forth in SEQ ID NO: A set of KASP (Kompetitive Allele Specific PCR) primers is provided for distinguishing one or more species from a species of the genus Ginseng containing three foliar ginseng and three lesions of lesion.

The term " competent allele specific PCR (KASP) " of the present invention is one of PCR-based analysis methods, homogenous and fluorescence-based genetic analysis techniques. KASP is a technique based on allele-specific oligo extension and fluorescence resonance energy transfer for signal generation.

The KASP primer set of the present invention is a primer set for detecting seven types of ginseng base types according to single nucleotide polymorphism (SNP) that is differentiated from left to right for ginseng species identification.

The primer set for KASP of the present invention is composed of two forward primers having a specific sequence corresponding to each SNP of a plant of the genus Ginseng in the same reverse primer (Primer_Common in Table 5) and thus having different fluorescent samples (Table 5 Primer_AlleleFAM / Primer_AlleleHEX) can be combined to identify ginseng species. For example, using SEQ ID NO: 48 (Primer_AlleleFAM) and SEQ ID NO: 47 (Primer_AlleleHEX) in the Commom primer, the product by the primer set of SEQ ID NO: 48 and the primer set of SEQ ID NO: (Blue) by SEQ ID NO: 48 and SEQ ID NO: 46 primer set because it is distinguished by HEX (green), SEQ ID NO: 48 and allele-specific primer SEQ ID NO: You can select Korean ginseng. Information on the other primer sets for KASP is shown in Table 5. The products of Commom primer and Primer_AlleleFAM among the respective primer sets represent FAM (blue), and each of the ginseng plants can be identified.

The dCAPS primer or the KASP primer according to the present invention may comprise 15 or more, 16 or more, 17 or more, 18 or more, 19 or more, 20 or more, An oligonucleotide consisting of at least 21, at least 22, at least 23, at least 24, at least 25, at least 26 consecutive nucleotides. For example, a primer (25 oligonucleotides) of SEQ ID NO: 10 may comprise at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, An oligonucleotide consisting of at least 22, at least 23, and at least 24 nucleotide fragments. The primers may also include additional, deleted, or substituted sequences of the nucleotide sequences of SEQ ID NO: 10 to SEQ ID NO: 99.

In the present invention, the term " primer " refers to a single strand of oligonucleotide sequence complementary to a nucleic acid strand to be duplicated, and may serve as a starting point for synthesis of a primer extension product. The length and sequence of the primer should allow the synthesis of the extension product to begin. The specific length and sequence of the primer will depend on the primer usage conditions such as temperature and ionic strength, as well as the complexity of the desired DNA or RNA target.

As used herein, an oligonucleotide used as a primer may also include a nucleotide analogue, such as phosphorothioate, alkylphosphorothioate, or peptide nucleic acid, or alternatively, And may include an intercalating agent.

In addition,

Isolating the genomic DNA from the ginseng sample;

Amplifying the target sequence by using the separated genomic DNA as a template and performing amplification reaction using the dCAPS primer set or the KASP primer set; And

And a step of analyzing the product of the amplification step to distinguish one or more species from the species of ginseng including Korean ginseng, American Samoan, Chinese ginseng, Korean ginseng, Vietnamese ginseng, Korean ginseng and lesion ginseng .

The method of the present invention comprises isolating genomic DNA from a ginseng sample. Methods for isolating genomic DNA from the ginseng sample can be performed by a method known in the art, and ethanol precipitation method following phenol: chloroform extraction may be used. Using the separated genomic DNA as a template, an oligonucleotide primer set according to an embodiment of the present invention may be used as a primer to amplify a target sequence by adding a DNA polymerase to perform an amplification reaction. Methods for amplifying a target nucleic acid include polymerase chain reaction (PCR), ligase chain reaction, nucleic acid sequence-based amplification, transcription-based amplification system, Strand displacement amplification or amplification with Q [beta] replicase, or any other suitable method for amplifying nucleic acid molecules known in the art. In the above, PCR is a method of amplifying a target nucleic acid from a pair of primers that specifically bind to a target nucleic acid using a polymerase. Such PCR methods are well known in the art, and commercially available kits may be used.

In the method of the present invention, the amplified target sequence may be labeled with a detectable labeling substance. In one embodiment, the labeling material can be, but is not limited to, a fluorescent, phosphorescent or radioactive substance. Preferably, the labeling substance may be FAM, HEX, VIC, JOE, ROX, TAMRA, Cy3 or Cy5. When the target substance is amplified, the 5'-end of the primer is labeled with the labeling substance and PCR is carried out, whereby the target sequence can be labeled with a detectable fluorescent substance. The set of oligonucleotide primers used to amplify the target sequence are as described above.

In the method according to an embodiment of the present invention, the analysis of the product of the amplification step may be performed by restriction enzyme cleavage, fluorescence measurement or phosphorescence measurement, but is not limited thereto.

The present invention also relates to the aforementioned dCAPS primer set or KASP primer set; And a reagent for carrying out an amplification reaction, wherein the kit comprises at least one species selected from the group consisting of Korean ginseng, American Samoan, Chinese medicinal herb, Korean white ginseng, Vietnamese ginseng, Korean ginseng and lesional ginseng .

In the kit of the present invention, the reagent for performing the amplification reaction may include a DNA polymerase, a DNA polymerase joinder, dNTPs, a buffer, and the like. In addition, the kit of the present invention may further include an internal control primer pair in addition to the primer set. In addition, the kit of the present invention may further include a user guide describing optimal reaction performing conditions. The manual is a printed document that explains how to use the kit, for example, how to prepare PCR buffer, the reaction conditions presented, and so on. The brochure includes instructions on the surface of the package including the brochure or leaflet in the form of a brochure, a label attached to the kit, and a kit. In addition, the brochure includes information that is disclosed or provided through an electronic medium such as the Internet.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example 1. Genomic sequence analysis and mutation analysis of chloroplasts of seven ginseng species to develop SNP markers

Sequences based on previous studies including the dnaLCW method were completed in order to compare and analyze the 7 chloroplast genomes of Korean ginseng, American ginseng, Chinese ginseng, Janseum japonica, Vietnamese ginseng, . Each chloroplast genome consists of four compartments, structurally including LSC, SSC, and two IRs. These chloroplast genomes commonly share 79 genes (CDS), 30 tRNAs, and 4 rRNAs Consists of. Each of the chloroplast genome sequences was multiplexed to search for and visualize the SNP mutation region, as shown in FIG.

A total of 1,783 SNPs were found among the seven chloroplast sequences of the genus Ginseng, of which 1,128 were located in the CDS region, and more SNPs were found in the LSC and SSC regions than in the IR region. The distribution pattern of SNPs was found in 72 of 79 CDSs based on the gene region. As a result , psaJ , psbN , rpl23 , psbF , psbL , rps18 and rps7 gene regions except these regions showed high retention rate It looked.

- Korean ginseng ( Panax ginseng ); American swan ( P. quinquifolius ); P. notoginseng ; P. japonicus ; Vietnam ( P. vietnamensis ); Panax stipuleanatus ; Trifolians ( P. trifolius ).

- The upper triangle represents the total number of SNPs observed when the chloroplast genomes of seven species of ginseng are compared one to one, and the lower triangle represents the number of SNPs in 79 CDS of each species.

There are few SNPs between Korean ginseng and American Sampler, which are diploid species compared to other species, and the variation of SNP between Samgwibae and Vietnam is the most common, and Samwipan has the most SNP compared to the other six species. The chloroplast genomes of ginseng plants have very high homology (≥ 97.6%), indicating that the chloroplast sequences are well preserved in ginseng and are consistent with the results for Araliaceae plants performed in previous studies can see. As mentioned above, the interspecific diversity among the chloroplast genomes was small and the interspecific diversity was high.

In addition, some genomes from the chloroplast to the mitochondria revealed transient sequences (Fig. 3). Twelve chloroplast genome fragments were transferred to mitochondria. These fragments varied from 2,297 bp to 8,250 bp, and more than 90% of each fragment sequence was identical. A marker was constructed with the exception of the 60,331 bp genomic region thus transformed. Using these results, a more accurate marker could be developed.

Therefore, we selected SNPs which are located in the genomic region and belong to the genomic region not transferred to mitochondria, and show specific nucleotide polymorphism in each of the seven ginseng genus as candidate markers. Whether a sequence similar to the surrounding sequences including the SNP is not present in the chloroplast genome, whether the GC content of the region to be primered is 20 to 80%, and whether the length of the product produced by the PCR reaction is not more than 250 bp And at least two species-specific SNP regions were selected for each species. Table 2 shows the SNP positions of the respective genes selected by the present invention and the names of the corresponding primers.

Gene
(SEQ ID NO) SNP location in gene dCAP  marker ID KASP  marker ID rpl20 (1) 249 Pgdm1 PgKm1 ndhK (2)

69 Pgdm2 PgKm2 187 Pndm9 PnKm9 213 Pqdm6 PqKm6 rps15 (3) 56 Pgdm3 PgKm3 ndhA (4) 310 Pqdm5 Pqkm5 rpoC1 (5)



996 X PjKm10 1044 Pqdm4 PqKm4 1362 Ptdm18 PtKm21 1658 Pndm7 PnKm7 1815 Psdm15 PsKm17 rPoC2 (6)

1263 Pvdm12 PvKm13 1320 Pndm8 PnKM8 2145 Pjdm10 X rPoB (7)
1158 Pjdm11 PjKm11 3186 Psdm16 PsKm18 ndhH (8) 171 Pvdm13 PvKm14 psbB (9) 414 Psdm14 PsKm16 ndhA (4) 831 Ptdm17 PtKm19

Example  2. Selected SNPs Marker  Based 18 oligonucleotides primer  Selection of dCAPS and KASP primer sets containing sets

Table 3 shows the names of the dCAPS primers and the KASP primers including the set of 18 oligonucleotide primers based on the SNP markers selected in the present invention and the SNP positions in these genes. Also, the respective sequences of the dCAPS primer and the KASP primer set are shown in Tables 4 and 5.

dCAPS  marker ID KASP  marker ID Gene SNP location in gene Pgdm1 PgKm1 rpl20 249 Pgdm2 PgKm2 ndhK 69 Pgdm3 PgKm3 rps15 56 Pqdm4 PqKm4 rpoC1 1044 Pqdm5 PqKm5 ndhA 310 Pqdm6 PqKm6 ndhK 213 Pndm7 PnKm7 rpoC1 1658 Pndm8 PnKm8 rpoC2 1320 Pndm9 PnKm9 ndhK 187 Pjdm10 x rpoC2 2145 x PjKm10 rpoC1 996 Pjdm11 PjKm11 rPoB 1158 Pvdm12 PvKm13 rpoC2 1263 Pvdm13 PvKm14 ndhH 171 Psdm14 PsKm16 psbB 414 Psdm15 PsKm17 rpoC1 1815 Psdm16 PsKm18 rPoB 3186 Ptdm17 PtKm19 ndhA 831 Ptdm18 PtKm21 rpoC1 1362

Abbreviation: Pg, Korean ginseng ( Panax ginseng ); Pq, American ginseng (P. quinquifolius); Pn, P. notoginseng ; Pj, P. japonicus ; Pv, Viet Nam ( P. vietnamensis ); Ps, Panax stipuleanatus ; Pt, Trifolius ( P. trifolius ).

Example 3. Screening of ginseng plants using a set of dCAPS primers comprising a set of 18 oligonucleotide primers based on the selected SNP markers

Each marker is a product separated into a set of dCAPS primers, and was designed to be divided into a restriction enzyme-cleaved sequence and a non-cleavable sequence according to the SNP upon band amplification (FIG. 4). Marker Pgdm 1 - 3 gene is located in the rpl20, ndhK and rps15 zone and is designed so that you can see the difference between the band length of Ginseng and other species are Ginseng specific marker. The marker Pqdm 4 - 6 is located in rpoC1 , ndhA , ndhK and is designed to have a band of specific length in the USA. Marker Pndm 7 - 9 is present in the rpoC1, rpoC2, and ndhK regions, and shows a different band from the other ginseng plants in the liver . Markers Pjdm 10 and 11 are located in rpoC2 and rpoB , respectively, and the markers Pvdm 12 and 13 have a specific band length from the rpoC2 and ndhH gene regions in Vietnam. Markers Psdm 14-16 are located in psbB, rpoC1 , and rpoB , and specifically act on the lesion three days. Markers Ptdm 17 and 18 are present in ndhA and rpoC1 , respectively. The band is amplified do. All 18 markers were successfully applied to distinguish seven ginseng plants and were therefore designed to identify seven species of ginseng.

This led to the development of 18 species-specific dCAPS markers in the SNP region between seven species of Panax ginseng. The SNPs of these markers are all located in the region of the protein-making gene, and they are specific to the genomes of all species among the chloroplasts of seven ginseng plant species. In addition, SNPs in regions where chloroplast sequences are similar to those of mitochondria are excluded due to genomic transfer to mitochondria.

Example 4. Selection of ginseng plants using a KASP primer set comprising 18 oligonucleotide primer sets based on the selected SNP markers

A fluorescent marker molecule marker was constructed in the SNP region of each marker except Pjdm10, and KASP (Kompetitive Allele Specific PCR) was performed using these markers (FIG. 5). The SNP region of the marker Pjdm10 was not suitable for producing a fluorescent marker molecule because allele in the genome was composed of three types. Therefore, the marker PjKm10 was further constructed by searching the SNP region located in rpoC2 in order to produce a fluorescent marker molecule capable of distinguishing the three types of pseudomonas.

Markers PgKm1 ~ PgKm3 are classified into different fluorescent markers according to the SNP between Korean ginseng and other species. The PqKm4 to PqKm6 make it possible to distinguish American watermelons as specific fluorescent markers by SNPs. The PnKm7 to PnKm9 markers target SNPs from the third and the last species, allowing them to be distinguished by specific fluorescent labels. Likewise, the PjKm10 to PjKm11 markers are distinguishable from other species through the fluorescent markers specific for the trichomes, and the PvKm13 to PvKm14 markers can distinguish them from other species through the specific fluorescent markers for the trichomes. In addition, the PsKm16 to PsKm18 markers can distinguish different species from other lesions with a specific fluorescence label of three days. The PtKm19 and PtKm21 markers can be amplified with specific fluorescent markers to differentiate them from other species. Using KASP markers produced from 18 SNPs, we constructed a classification system that has good visibility and reproducibility and is easy to use for analysis.

<110> Seoul National University R & DB Foundation <120> SNP Marker derived from complete sequencing of chloroplast genome          of seven Panax species, primer set for discrimination of Panax          species and uses thereof <130> PN18076 <160> 99 <170> Kopatentin 2.0 <210> 1 <211> 387 <212> DNA <213> Panax spp. <400> 1 atgaccagaa ttagacgggg atatatagct cggagacgta gaacaaaaat tcgtttattt 60 gcatcaagct ttcgaggggc tcattcaagg cttactcgaa caattactca acagaaaata 120 agagctttgg tttcgtctca tcgggatagg gatcggcaaa agagaaattt tcgtcgtttg 180 tggatcactc gaataaacgc agtaattcgt gaaggggggg tatcctatag ttatagtaga 240 ttaatacatg atctgtataa gaaacagtta cttcttaatc gtaaaatact tgcacaaata 300 gctatatcaa ataaaaattg tctttatatg atttccaatg agatcataaa agaagtagat 360 tggaaagaat ccaccggaat aatttaa 387 <210> 2 <211> 678 <212> DNA <213> Panax spp. <400> 2 atgaattcca tcgagtttcc cttacttgat cgaacaaccc aaaattcagt tatttcaact 60 acatcaaatg atctttcaaa ttggtcaaga ctctctagtt tatggccgct tctctatggt 120 actagttgtt gcttcattga atttgcttca ctaataggct cacgattcga ctttgatcgt 180 tatggactag tcccaagatc gagtcctaga caagcggacc taattttaac agccggaaca 240 gtaacaatga aaatggctcc ttctttagtg agattatatg agcaaatgcc tgaaccaaaa 300 tatgttattg ctatgggagc atgtacaatt tcagggggga tgttcagtac tgattcttat 360 agtactgttc ggggagtcga taagctaatt cctgtcgatg tctatttgcc aggctgtcca 420 cctaaacccg aagcagttat agatgctata acaaaacttc gtaaaaaaat atctcgagaa 480 atctatgaag atagaattaa gtctcaacgg gagaatcggt gttttacaac caatcacaag 540 tttcaggttg ggcgtagtat tcatactgga aattatgatc gagggttcct ttatcaaccg 600 ccatttactt cagagatccc ttccgaaaca tttttcaaat acaaaagttc agtctcgtcc 660 cacgaattag tgaattag 678 <210> 3 <211> 273 <212> DNA <213> Panax spp. <400> 3 atgataaaaa atccattcag tggaattatt ttgaaagagg aaaacaaaga caacaggggg 60 tctgctgaat ttcaagtagt cagtttcacc aataggatac gaagacttac ttcacatttg 120 gaattgcaca aaaaagacta tttatctcag agaggtctgc gtaaaattct aggaaagcgt 180 caacggctac tggcttattt gtcaaaaaaa aatagagtac gttataaaga attaattggt 240 cggttggata ttcgagaaac aaaaattcgt taa 273 <210> 4 <211> 1092 <212> DNA <213> Panax spp. <400> 4 atgataatag atacaacaga agtacaagct atcaattctt tttttagatt ggaatcctta 60 aaagaggtat atgggatcat atggatgctt atccctattt ttactcctgt attaggaatc 120 acaataggtg tactagtaat tgtttggtta gaaagagaaa tatctgcagg gatacaacaa 180 cgtattggac ctgaatacgc cgggcctttg ggaattcttc aagctttagc agatggtaca 240 aaattacttt tcaaagagaa tcttcttcca tcgagaggag atactcgttt attcagtatc 300 ggaccatccg tagcagttac atcaattcta ctaagttatt tagtaattcc ttttggctat 360 caccttgttc tagccgatct cagtatcggt gtttttttat ggattgccat ttcaagtatt 420 gctcccgttg gccttcttat gtcaggctat ggatcaaata ataaatattc ctttttaggt 480 ggtttacgag ctgctgctca atcaattagt tatgaaatac cattaactct atgtgtgtta 540 tcaatatctc tattatctaa cagttcaagt acagttgata tagttgaagc gcagtcaaaa 600 tatggttttt gggggtggaa tttgtggcgt caacctatag ggtttatcat ttttctaatt 660 tcgtccctag ccgagtgtga gagattacct tttgatttac cagaagcaga agaagaattg 720 gtagcaggtt atcaaaccga atattcaggt ataaaatttg gtttatttta cgttgcttcg 780 tatctgaatc tactagtttc ttcattattt gtaacggttc tttacttggg gggttggaat 840 ctttctattc cgtacatacc cgttcctgat atttttgaaa taaataaagc gagtagagta 900 tttggaacaa taactggtat ctttattaca ttagctaaaa cttatttgtt cttattcatt 960 cctatcacaa caagatggac tttaccaagg ctaagaatgg accaactatt aaatcttgga 1020 tggaaatttc ttttacccat ttctctcggc aatctattat taacaacctc ttcccaactt 1080 ctttcactgt aa 1092 <210> 5 <211> 2070 <212> DNA <213> Panax spp. <400> 5 atgaatcaga atttttcttc tatgatcgat cggtataaac atcaacaact ccgaattgga 60 tcagtttctc ctcaacaaat aagtacttgg gccaataaaa tcctgcccaa tggagagata 120 gttggagagg tgacaaaacc ttatactttt cattacaaaa ccaataaacc ggaaaaagat 180 ggattatttt gtgaaagaat ttttgggcct atcaaaagcg gaatttgtgc ttgtggaaat 240 tatcgagtaa tcgggaatga aaaagaagac ccgaaatttt gtgaacaatg cggagtcgaa 300 tttgttgatt ctcggatacg aagatatcaa atgggctaca tcaaactcgc atgcccagta 360 acccatgtgt ggtatttgaa acgtcttcct agttatattg cgaatctttt agataaacct 420 cttaaagaat tagaaggcct agtatactgc gatttttctt ttgcgaggcc catagctaaa 480 aaacctactt tcttacgatt acgaggttcg ttcgaatatg aaatccaatc ctggaaatac 540 agcatcccgc ttttttttac tacccaaggc ttcgatacat ttcgaaatcg agaaatttct 600 actggagcag gtgctatccg agaacaatta gccgatctcg atttacggat tattatagat 660 tcttcgttgg tagaatggaa agagttgggg gaagacgggc ctacagggaa tgaatgggaa 720 gatcgaaagg ttggaagacg aaaggatttt ttggttagac gtatggaatt agctaagcat 780 tttattcgaa caaatataga accagaatgg atggttttgt gtctattacc tgttcttcct 840 cctgagttga gaccgatcat tcagatagat gggggtaaac taatgagctc agatatta 900 gaactctata gaagagttat ctatcgaaac aatactctta ccgacctatt aacaacaagt 960 agatcaacgc caggagaatt agtaatgtgt caggagaaat tggtacaaga agccgtggat 1020 acacttcttg ataatggaat ccgcggacaa ccgatgaggg acggtcataa taaagtttac 1080 aagtcatttt cagatgtaat tgaaggcaaa gagggaagat ttcgtgagac tctgcttggc 1140 aaacgggtcg attattcagg gcgttccgtc attgtcgtgg gtccgtcact ttcattacat 1200 cgatgtggat tgccccgcga aatagcaata gagcttttcc agacatttgt aattcgtggt 1260 ctaattagac aacatcttgc ttcgaacata ggagttgcta agagtaaaat tcgggaaaaa 1320 gaaccgattg tatgggaaat acttcaggaa gttatgcagg ggcatcccgt attgctaaat 1380 agagcgccga ctctgcatag attaggcata caggcattcc agcctgtttt agtggaaggt 1440 cgtgctattt gtttacatcc attagttcgt aaaggattca atgcagactt tgacggggat 1500 caaatggctg ttcatgtgcc tttatctttg gaggctcaag cagaagcacg tttacttatg 1560 ttttctcata tgaacctttt gtctccagct attggggatc ctatttccgt accaactcaa 1620 gatatgctta ttggactcta tgtcttaacg agcgggaatc gtcggggtat ttgtgtaaat 1680 aggtataatc catgtaatcg cagaaactat caaaatgaaa gaatttacga taataactat 1740 aagtatacga aaaaaaaaga accctttttt tgtaattcct atgatgcaat tggagcttat 1800 cggcagaaac gaatcaattt agatagtcct ttgtggcttc ggtggcgact agatcaacgc 1860 gttatttctg caaaagaagc ccccctcgaa gttcactatg aatctttggg tacctattat 1920 gaaatttatg ggcaatatct aatagtaaga agtataaaaa aagaaattct atatatatat 1980 attcgaacta ctgttggtca tatttctctt tatcgagaaa tcgaagaagc tatacagggg 2040 ttttgtcagg cctgttcata tggtccctaa 2070 <210> 6 <211> 4164 <212> DNA <213> Panax spp. <400> 6 atggaggtac ttatggcaga acgggccaat ctggtctttc acaataaagt gatagacgga 60 actgccatga aacgacttat tagtagatta atagatcact tcggaatggc atatacatca 120 cacatcctgg atcaagtaaa gactttgggt tttcaacaag ctactgctac ctccatttca 180 ttaggaattg acgatctttt aacaatccct tctaagagat ggctagttca agatgctgaa 240 caacaaagtt taattttgga aaaacaacac cattatggga atgtacacgc ggtagaaaaa 300 ttacgtcaat ccattgagat atggtatgct acaagtgaat ttttgcgaca agaaatgaat 360 cctaatttta ggatgactga cccctttaat ccagttcata taatgtcttt ttcgggagct 420 agaggaaatg catctcaggt acatcaatta gtaggtatga gaggattaat ggcggatccc 480 caaggacaaa tgattgattt acccattcaa agcaatttac gcgaaggact ctctttaaca 540 tgtacgaacc 600 tcagatgctg gatatctcac gcgcagactt gttgaagtag ttcaacacat tgttgtacgt 660 cgaaaagatt gtggcaccgt ccgaggtatt tctgtgagtc cccgaaatgg gatgatgccg 720 gaaagggttt ttatccaaac attaattggt cgtgtattag cggatgatat atatctgggt 780 ccacgatgta ttgccaccag aaatcaagat attggtagtg gacttgtcaa tcgattcata 840 acctttcgag cacaaccaat atatattcga actcccttta cttgtaggag tacatcttgg 900 atctgtcgat tatgttatgg ccggagtcct actcacggcg atctggtcga attgggagaa 960 gctgtgggta ttattgcagg tcaatctatt ggagaacctg gtactcaatt aacattaaga 1020 acctttcata ccggcggagt attcacaggg ggtactgcag aacatgtgcg agccccttct 1080 aatggaaaaa taaaattcaa tgaggatttg gttcatccga cacgtacacg tcatggacat 1140 cctgcttttc tatgttctat agacttgtat gtaactattg agagtgaaga tattatacat 1200 aatgtgaata ttccacctaa aagttttctt ttagttcaaa atgatcaata tgtagaatca 1260 gaacaagtga ttgctgagat tcgcgcagga acatccactt tgaattttaa agagaaggtt 1320 cgaaaacata tttattctga ctcagaggga gaaatgcact ggaataccga tgtgtatcat 1380 gcacctgcat ttacttatgg gaatgttcat ctcttaccaa aaacaagtca tttatggata 1440 ttattaggag agccatgcag atccgatcta gtcgccctct cgatccacaa ggatcaagat 1500 caaatgaacg cccattctct ttctgtcaag agaagatata tttctaacct ctcagtaact 1560 aatgatcaag cgagacacaa attctttagt tcggattttt ctggtaaaaa agaagatagg 1620 attactgatt attcagaact taatcgaatt ggtcattgta atctcccata tctggccatt 1680 ctccacgcaa attctgattt attggcaaag agacgaagaa atagattcat cattccactc 1740 caatcgattc aagaacacga gaacgaacta atgccctctt cgggtatctc gattgaaata 1800 cccatacatg gtattttccg taaaaagagt attattgctt atttcgatga tcctcgatac 1860 agaagaaaga gttcgggaat tactaaatat ggtactatag aagtgcattc aatcgtcaaa 1920 aaagaggatt tgattgagta tcgaggagtc aaggaattta ggccaaaata ccaaatgaaa 1980 gtggatcgat tctttttcat tcccgaggaa gtgcatatct tacccggatc ttcttccata 2040 atggtacgga acaatagtat tattggggta gatacacaaa tcactttaac tacaagaagc 2100 cgagtgggcg gattggtccg agtggagaga aaaaaaaaaa gaattgaact taaaatattt 2160 tctggagata tccattttcc tggagagacg gataagatat cccgacatag tggcgtcttg 2220 ataccaccag gaaccggaaa aacaaattcc aaggaatcaa aaaaatggaa aaattggatc 2280 tatgtccaac ggatcacacc taccaagaaa aagtattttg ttttggttcg acctgtagtc 2340 acatatgaaa taacggacgg tataaattta gcaacacttt tcttcccgga tctgttgcag 2400 gaaagggata atgtgcaact tcgagttgtc aattatatcc tttatggaac tggcaaacca 2460 attcggggtt tttatgacac aagtattcaa ttagttagga cttgtttagt attaaattgg 2520 gtccaagaca aaaaaagttc ttctatcgaa gaggcccgta cttcctttgt tgaaataagg 2580 ataaatagtt tgattcgaga ttttctaaaa atcgacttag caaaatcccc tattttgtat 2640 accggaaaaa ggaaagatcc atcgggttca ggattgatct ctgagaatgg atcagatcgc 2700 accaatatca atccgttttc ttccagtttt ttctattcca aggcaaggat taaagaatcc 2760 cttaatcaaa accaaggaac tattcataca ttgttgaata gaaataagga atgccaatct 2820 ttgataattt tgtcatcatc caattgtttt cgaatgggtc cattcaacga tgtaaaatat 2880 cacaatgtga taaaagaatc aattcaaaaa gaccccctaa ttcaaattag gaattcgttg 2940 ggccctttag gaacggccct tcaaattgcg aatttttatt cattttccca tttaataact 3000 tacaatcaga tcctggtaac taactatttg caacttgaca atttaaaaca gagttttcaa 3060 gtacttaaat attatttaat ggatgaaaat gggaaaattt ataatcccga tcgatgctgt 3120 aacattattt tgaatccatt caatttgaat tggttttttc tccatcacaa ttattgtgaa 3180 gagacgtcta caacaatgag tcttgggcag tttatttgtg aaaatgtatg tatagccaaa 3240 aatggaccac acctaaaacc gggtcaagtt ctaattgttc aagttgactc tgtagtaata 3300 cgatccgcta agccttattt ggccacccca ggagcaactg ttcatggcca ttatggggaa 3360 atcctttacg aaggagatac attagttaca tttatatatg aaaaatcgcg atctggtgat 3420 ataacgcagg gtcttccaaa agtcgaacag gtgttagaag tgcgttcgat tgattcaata 3480 tcgatgaatc tcgaaaagag ggttgagggc tggaacgaat gtataacaag aaatcttgga 3540 attccttggg gattcttgat tggggctgag ctaactatag tgcaaagtcg tatttctttg 3600 gttaataaga tccaaaaggt ttatcgatcc cagggggtgc agatccataa taggcatata 3660 gaaattattg tacgtcaaat aacatcaaaa gtgttggttt cagaagatgg gatgtctaat 3720 gtttttttac ccggagaatt tattggattg ttgcgagcgg aacgaatggg tcgcgctttg 3780 gaagaagcga tctgttacca agccgtctta ttgggaataa caaaagcatc tctgaatact 3840 caaagtttca tatccgaagc gagttttcaa gaaactgctc gagttttagc aaaagcagct 3900 ctccggggtc gtatcgattg gttgaaaggc ctgaaagaga acgttgttct ggggggaatg 3960 atacccgttg gtaccggatt caaaggattg gtgccccctt cgaggcaaca taagaatatt 4020 cccttggaaa ccaaaaataa gaatctattc gagggggaaa tgagagatat tttgttccac 4080 cacagaaaat tttttgattc ttgcctttca aagaatttcc atgatacacc agaacaatca 4140 tttataggat ttaatgattc ctaa 4164 <210> 7 <211> 3216 <212> DNA <213> Panax spp. <400> 7 atgctccgag atggaaaaaa tgagggaatg tctacaatac ctgggtttaa tcagatacaa 60 tttgaaggat tttgtaggtt cattgatcag ggcttgacgg aagaacttta taagtttcca 120 aaaattgaag ataccgacca agaaattgaa tttcaattat ttgtggaaac atatcaatta 180 gtagaaccgt tgataaaaga aagagatgct gtgtatgaat cactcacata ttcttctgaa 240 ttatatgtct ccgcgggact catttggaaa actggtaggg atatgcaaga acaaactatt 300 ttttttggaa acattcctct aatgaattct ctgggaactt ctatagtaaa tggaatatat 360 agaattgtga tcaatcaagt attgcaaagc cccggtattt attaccggtc agaattggac 420 cataacggaa tttcggtgta taccggcacc ataatatcag attggggagg aagatcagaa 480 ttagagattg atagaaaagc aaggatatgg gctcgtgtga gtaggaaaca aaaaatatct 540 attctagttt tatcatcagc tatgggttcg aatctaagag aaattctaga gaatgtttgc 600 tatcctgaaa tatttttgtc ttttctgact gataaggaga aaaaaaaaat cgggtcaaaa 660 gaaaatgcca ttttggagtt ttaccaacaa tttgcttgtg taggcgggga tccggtattt 720 tctgaatcct tatgtaagga attacaaaag aaattctttc aacaaagatg tgaattagga 780 aggattggtc gacgaaatat gaatcggaga ctgaaccttg atatatccca gaacaataca 840 tttttgttac cgcgagatat attggcagcc gcggatcatt tgattggaat gaaatttgga 900 atgggtacac ttgacgatat gaatcatttg aaaaataaac gtattcgttc tgtagcagat 960 cttttacaag atcaattcgg attggctctg gttcgtttag aaaatgtggt tcgaggaact 1020 atatgtggag cacttaggca taaattgata ccgactcctc agaatttggt aacttcaact 1080 ccattaacaa ctacttatga atcttttttc ggtttacacc cattatctca agttttggat 1140 cgaactaatc cattgacaca aatagttcat gggagaaaat tgagttattt gggccctgga 1200 ggattgacag ggcgaactgc tagttttcga atacgagata tccatcctag tcactatggg 1260 cgtatttgcc caattgacac gtctgaagga ataaatgtcg gacttattgg atctttagca 1320 attcatgcga ggattggtcg ttggggatct ctagaaagcc cgttttatga aatttctgag 1380 agatcaaaag gggcacggat gctttattta tcaccaggta aagatgaata ctatatggta 1440 gcggcgggaa attctttggc cttgaatcag ggtattcagg aagaacaggt tgttccagct 1500 cgataccgtc aagaattcct gactattgcg tgggaacagg ttcatcttcg aagtattttt 1560 tccttccaat atttttctat tggagcttcc ctcattcctt ttatcgagca taatgatgcg 1620 aatcgggctt taatgagttc gaatatgcaa cgtcaagcag ttcctctttc tcggtccgag 1680 aagtgcattg ttggaactgg gttagaacgc caagcggcta tagattcagg ggctcttgct 1740 atagcggaac acgagggaaa gatcatttat accgatactg acaagatcct tttatcaggt 1800 aatggaaata ctctaagcat tccattagtt atatatcaac gctccaacaa aaatacttgt 1860 atgcatcaaa aaccccaggt tcagcggggt aaatgcataa aaaagggaca aattttagcc 1920 catggcgccg ctacagttgg tggcgaactc gctttgggga aaaacgtatt agtagcttat 1980 atgccatggg aaggttacaa ttttgaagat gcagtactca ttagcgagcg cttggtatat 2040 gaagatattt atacttcttt tcacatacga aaatatgaaa ttaagactca tgtgacaagc 2100 caaggccccg aaagggtcac taatgaaata ccgcatttag aagcccattt actccgcaat 2160 ttagacaaaa acggaattgt aatgctggga tcttgggtag agacgggtga tattttagta 2220 ggtaaattaa cgccccaaat ggtgaaagaa tcgtcgtatg ccccggaaga tagattgtta 2280 cgagccatac ttggcattca ggtatctact tcaaaagaaa cttgtctaaa actacctata 2340 ggtggtaggg gtcgggttat tgatgtgaga tggatccaga aaaggggagg ttctagttat 2400 aacccagaaa tgattcgtgt atatatttca cagaaacgtg aaattaaggt aggcgataaa 2460 gtagctggaa ggcatggaaa taagggtatc atttcaaaaa ttttgcctag acaagatatg 2520 ccttatttgc aagatggaag acctgttgat atggtcttca acccattagg agtaccttca 2580 cgaatgaatg tagggcagat atttgaatgt tcactcgggt tagcgggggg tctgctagac 2640 agacattatc gaatagcgcc ttttgatgag agatatgaac aagaagcttc gagaaaacta 2700 gtgttttctg aattatatga agccggtaag caaacagcga atccatgggt atttgaaccc 2760 gagtatccgg gaaaaagtag aatatttgat ggaagaacgg gggatccttt tgaacaacct 2820 gttataatag gaaagcctta tatcttgaaa ttaattcatc aagttgatga taaaatccat 2880 ggacgttcca gcggacatta tgcacttgtt acacaacaac cccttagagg aagggccaag 2940 caaggtggac aacgggtagg agagatggag gtttgggctc tagagggatt tggtgttgct 3000 catattttgc aagagatgct tacttataaa tcggatcata ttagagctcg ccaggaagta 3060 cttggtacta cgatcattgg aggaacaata cctaatcctc aggatgctcc agaatctttt 3120 cgattgctcg ttcgagaact acgatctttg gctctggaac tgaatcattt ctttgtatct 3180 gagaagaatt tccagattaa taggaaggaa gcttaa 3216 <210> 8 <211> 1182 <212> DNA <213> Panax spp. <400> 8 atgactgcat cagctacacg aaatgacctt atgatagtca atctgggtcc tcagcaccca 60 tcaatgcacg gtgttcttcg actcattgtt actctagatg gtgaagatgt tattgactgt 120 gaaccaatat tgggttattt acatagaggg atggaaaaaa ttgcggaaaa ccgaacaatt 180 atacagtatt taccttatgt aacacgttgg gattatttag ctactatgtt cacagaagca 240 ataactgtaa atgcaccaga gcagttaggc aatattcaag tacctaaaag ggccagctat 300 atcagagtca ttatgttgga gttaagtcgt atagcttcgc atttgttatg gcttggcccc 360 tttctggcag atattggcgc acagacccct ttcttctata tttttcgaga aagagaattg 420 atatatgacc tattcgaagc tgccaccggt atgcgaatga tgcataatta ttttcggatc 480 ggaggagtag ctgctgattt acctcatgga tggatagata aatgtttgga tttttgtgat 540 ttttttttaa caagggttac tgaatatcaa aaacttatta cacggaaccc tattttttta 600 gaacgggttg aaggagtagg cattattggc ggagcggagg caataaattg gggtttatca 660 ggaccaatgc tacgagcttc cggaatacaa tgggatcttc gtaaagttga tcattatgag 720 ttttacgacg aatttgattg gggggtccaa tggcaaaaag agggggattc cttagctcgt 780 tatttagtac gaatcagtga aatgacagaa tccataaaaa taattcaaca ggctttagaa 840 ggaattccgg gggggcccta tgagaattta gaaatccgac gttttgataa agtacgggat 900 cctgaatgga atgattttga ctatcgattc atcagtaaaa aaccttctcc gacttttgaa 960 ttatcaaagc aagaacttta tgtgagagtc gaagccccca agggagaatt ggggattttt 1020 ctgatagggg ataggggtgt ttttccttgg agatggaaaa tccgaccacc gggttttatc 1080 aatttgcaaa tccttcctca gttagttaaa agaatgaaat tggctgatat tatgacgata 1140 ctaggtagca tagatataat tatgggagaa gttgatcgtt aa 1182 <210> 9 <211> 1527 <212> DNA <213> Panax spp. <400> 9 atgggtttgc cttggtatcg tgttcatacc gttgtattga atgatcccgg ccggttgctt 60 tctgtccata taatgcatac agctctagtt gctggttggg cgggttcgat ggctctatat 120 gaattagcag tttttgatcc ctctgaccct gttcttgatc caatgtggag acagggtatg 180 ttcgttatac ccttcatgac tcgtttagga ataaccaatt catggggcgg ttggagtgtc 240 acaggagggg ctataccgaa tccgggtatt tggagttacg aaggtgtggc cggggcacat 300 attgtgtttt ctggcttgtg cttcttggca gctatctggc attgggttta ttgggatcta 360 gaaatttttt ctgatgaacg tacaggaaaa ccttctttgg atttgcccaa gatctttgga 420 attcatttat ttctcgcagg ggtggcttgc tttggttttg gtgcatttca tgtaacaggc 480 ttgtatggtc ctggaatatg ggtgtctgat ccttatggac taacgggaaa agtacaatct 540 gtaaatccag cgtggggtgt ggaaggtttt gatccttttg ttccgggagg aatagcctct 600 catcatattg cagcagggac attgggtata ttggccggtc tattccatct tagtgtccgc 660 ccgccccaac gtctatacaa aggattgcgt atgggcaata ttgaaaccgt cctttccagt 720 agtatcgctg ctgtcttttt tgcagctttt gttgttgccg gaactatgtg gtatggttca 780 gcaactaccc cgatcgaatt atttgggcct actcgttatc aatgggatca ggggtacttc 840 cagcaagaga tatatcgaag agttagtgct gggctagccg aaaatcaaag tttatcagaa 900 gcttggtcta aaattcctga aaaattagct ttttatgatt acatcggcaa taatccggca 960 aaagggggat tattcagagc gggttcaatg gataacgggg atggaatagc ggttggatgg 1020 ttaggacatc ctatctttag agataaagaa gggcgtgaac tttttgtacg tcgtatgcct 1080 accttttttg aaacctttcc ggtcgttttg gtagatggcg acggaattgt tagagccgat 1140 gttccttttc gaagggcaga atcgaagtat agtgtcgaac aagtaggtgt aactgttgag 1200 ttctacggcg gcgaactcaa cggagtcagt tatagtgatc ctgctactgt gaaaaaatat 1260 gctagacgtg ctcaattggg tgaaattttt gaattagatc gtgctacttt gaaatccgat 1320 ggtgtttttc gtagcagtcc gaggggttgg tttacttttg gacatgcttc ttttgctttg 1380 ctcttctttt ttggacatat ttggcatggt gctagaacct tgttcagaga tgtttttgct 1440 ggtattgatc cagatttgga tgctcaagta gaatttggag cattccaaaa actgggagat 1500 ccaactacaa gaagacaagt agtctga 1527 <210> 10 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 10 gtttaaatta ttccggtgga ttctt 25 <210> 11 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 11 gtagcctata gttatagtag attaatcga 29 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 12 gtccgcttgt ctaggactcg 20 <210> 13 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 13 caaaattcag ttatttcaac tacatcaat 29 <210> 14 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 14 atccaaccga ccaattaatt cttta 25 <210> 15 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 15 ttgaaagagg aaaacaaaga caccc 25 <210> 16 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 16 tatgaccgtc cctcatcggt tgtcg 25 <210> 17 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 17 cattcagata gatgggggta aacta 25 <210> 18 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 18 ctcgtaaacc acctaaaaag gaat 24 <210> 19 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 19 tcgtttattc agtatcggac catcg 25 <210> 20 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 20 ttccggctgt taaaattagg tcagc 25 <210> 21 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 21 tctttcaaat tggtcaagac tctct 25 <210> 22 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 22 cctatttaca caaatacccc gtcga 25 <210> 23 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 23 attagttcgt aaaggattca atgcag 26 <210> 24 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 24 ttcatttgat cttgatcctt gtg 23 <210> 25 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 25 tccactttga attttaaaga gaagct 26 <210> 26 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 26 atcgacagga attagcttat cgac 24 <210> 27 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 27 acgattcgac tttgatcgtt atcga 25 <210> 28 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 28 tggatatctc cagaaaatat tttaagtac 29 <210> 29 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 29 aggatttgat tgagtatcga ggag 24 <210> 30 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 30 agtccgacat ttattccttc agac 24 <210> 31 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 31 gtttggatc gaactaatcc attggt 26 <210> 32 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 32 tgcgcgaatc tcagcaatca ctag 24 <210> 33 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 33 aaattcaatg aggatttggt tcat 24 <210> 34 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 34 cataaggtaa atactgtata attgatcg 28 <210> 35 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 35 tatgatagtc aatctgggtc ctca 24 <210> 36 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 36 aaccttcttt ggatttgccc aagct 25 <210> 37 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 37 cacgctggat ttacagattg tact 24 <210> 38 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 38 gaagccacaa aggactatct aaatg 25 <210> 39 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 39 gtcggggtat ttgtgtaaat aggt 24 <210> 40 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 40 taagcttcct tcctattaat ctgggaatt 29 <210> 41 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 41 catattagag ctcgccagga agta 24 <210> 42 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 42 tatgtacgga atagaaagat tccaagc 27 <210> 43 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 43 cgagtgtgag agattacctt ttga 24 <210> 44 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 44 cgctctattt agcaatacgg gata 24 <210> 45 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 45 gcaatagagc ttttccagac attt 24 <210> 46 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 46 gattaagaag taactgtttc ttatacagat ca 32 <210> 47 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 47 aagaagtaac tgtttcttat acagatcg 28 <210> 48 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 48 ggggggtatc ctatagttat agtagatta 29 <210> 49 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 49 gagagtcttg accaatttga aagatca 27 <210> 50 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 50 agagtcttga ccaatttgaa agatcg 26 <210> 51 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 51 tcccttactt gatcgaacaa cccaaaatt 29 <210> 52 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 52 attttgaaag aggaaaacaa agacaacag 29 <210> 53 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 53 aattattttg aaagaggaaa acaaagacaa caa 33 <210> 54 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 54 tcgtatccta ttggtgaaac tgactactt 29 <210> 55 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 55 gatacacttc ttgataatgg aatccgt 27 <210> 56 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 56 atacacttct tgataatgga atccgc 26 <210> 57 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 57 ctttattatg accgtccctc atcggtt 27 <210> 58 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 58 aaataactta gtagaattga tgtaactgct at 32 <210> 59 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 59 ataacttagt agaattgatg taactgctac 30 <210> 60 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 60 agatactcgt ttattcagta tcggaccat 29 <210> 61 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 61 cccaagatcg agtcctagac ag 22 <210> 62 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 62 cccaagatcg agtcctagac aa 22 <210> 63 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 63 cattttcatt gttactgttc cggctgtta 29 <210> 64 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 64 atttacacaa ataccccgac gac 23 <210> 65 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 65 cctatttaca caaatacccc gacgat 26 <210> 66 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 66 ccaactcaag atatgcttat tggactcta 29 <210> 67 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 67 ctgagtcaga ataaatatgt tttcgg 26 <210> 68 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 68 cctctgagtc agaataaata tgttttcga 29 <210> 69 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 69 cgcgcaggaa catccacttt gaatt 25 <210> 70 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 70 acgattcgac tttgatcgtt atggat 26 <210> 71 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 71 cgattcgact ttgatcgtta tggac 25 <210> 72 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 72 tgtctaggac tcgatcttgg gacta 25 <210> 73 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 73 caggagaatt agtaatgtgt caggaa 26 <210> 74 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 74 caggagaatt agtaatgtgt caggag 26 <210> 75 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 75 gtgtatccac ggcttcttgt accaa 25 <210> 76 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 76 caattttctc ccatgaacta tttgc 25 <210> 77 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 77 aactcaattt tctcccatga actatttgt 29 <210> 78 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 78 cccattatct caagttttgg atcgaacta 29 <210> 79 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 79 gttcaaaatg atcaatatgt agaatcagag 30 <210> 80 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 80 agttcaaaat gatcaatatg tagaatcaga a 31 <210> 81 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 81 atgttcctgc gcgaatctca gcaat 25 <210> 82 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 82 tacataaggt aaatactgta taattgttcg a 31 <210> 83 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 83 acataaggta aatactgtat aattgttcgg 30 <210> 84 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 84 catagaggga tggaaaaaat tgcggaaaa 29 <210> 85 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 85 aaaccttctt tggatttgcc caagatt 27 <210> 86 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 86 accttctttg gatttgccca agatc 25 <210> 87 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 87 acccctgcga gaaataaatg aattccaaa 29 <210> 88 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 88 ggagcttatc ggcagaaacg aatt 24 <210> 89 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 89 gagcttatcg gcagaaacga atc 23 <210> 90 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 90 caccgaagcc acaaaggact atcta 25 <210> 91 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 91 aagcttcctt cctattaatc tggaaattt 29 <210> 92 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 92 gcttccttcc tattaatctg gaaattc 27 <210> 93 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 93 ggctctggaa ctgaatcatt tctttgtat 29 <210> 94 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 94 cattatttgt aacggttctt tacttggga 29 <210> 95 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 95 atttgtaacg gttctttact tgggg 25 <210> 96 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 96 cggatatgta cggaatagaa agattccaa 29 <210> 97 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 97 cgctctattt agcaatacgg gatgt 25 <210> 98 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 98 gctctattta gcaatacggg atgc 24 <210> 99 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 99 cgattgtatg ggaaatactt caggaagtt 29

Claims (7)

A single nucleotide polymorphism (SNP) located at the 249th base in the nucleotide sequence of the rpl20 ( ribosomal protein L20 ) gene of SEQ ID NO: 1 in Panax ginseng; SNP located at the 69th base, 187th and 213th bases of the nucleotide sequence of ndhK ( NADH-plastoquinone oxidoreductase subunit K ) of SEQ ID NO: 2; A SNP located at the 56th base of the nucleotide sequence of the rps15 ( ribosomal protein S15 ) gene of SEQ ID NO: 3; SNP located at the 310th and 831st bases of the nucleotide sequence of ndhA ( NADH-plastoquinone oxidoreductase subunit 1 ) gene of SEQ ID NO: 4; SNPs located at the 996th , 1044th , 1362th , 1658th , and 1815th bases of the nucleotide sequence of the rpoC1 ( RNA polymerase beta subunit ) gene of SEQ ID NO: 5; SNPs located at bases 1263, 1320 and 2145 of the nucleotide sequence of rpoC2 ( RNA polymerase beta subunit ) of SEQ ID NO: 6; SNPs located at the 1158th and 3186th bases of the nucleotide sequence of the rpoB ( RNA polymerase beta subunit ) gene of SEQ ID NO: 7; A SNP located at nucleotide 171 in the nucleotide sequence of ndhH ( NADH-plastoquinone oxidoreductase subunit 7 ) of SEQ ID NO: 8; And a SNP located in the 414 base sequence of the nucleotide sequence of psbB ( photosystem II CP47 chlorophyll apoprotein ) gene of SEQ ID NO: 9, or a complementary polynucleotide of the polynucleotide composed of 8 or more consecutive nucleotides, Panax ginseng , P. quinquifolius , P. notoginseng , P. japonicus , P. vietnamensis , P. trifolius and Panax stipuleanatus , A single nucleotide polymorphism (SNP) marker composition for distinguishing one or more species from a ginseng species. SEQ ID NOS: 10 and 11; SEQ ID NOS: 12 and 13; SEQ ID NOS: 14 and 15; SEQ ID NOS: 16 and 17; SEQ ID NOS: 18 and 19; SEQ ID NOS: 20 and 21; SEQ ID NOS: 22 and 23; SEQ ID NOS: 24 and 25; SEQ ID NOS: 26 and 27; SEQ ID NOS: 28 and 29; SEQ ID NOS: 30 and 31; SEQ ID NOS: 32 and 33; SEQ ID NOS: 34 and 35; SEQ ID NOS: 36 and 37; SEQ ID NOS: 38 and 39; SEQ ID NOS: 40 and 41; SEQ ID NOS: 42 and 43; And oligonucleotide primers set forth in SEQ ID NOS: 44 and 45, and a primer set of SEQ ID NOs: 44 and 45. The primers include dCAPS (derived cleaved amplified polymorphic sequence) primer set. SEQ ID NOS: 46, 47 and 48; SEQ ID NOS: 49, 50 and 51; SEQ ID NOS: 52, 53 and 54; SEQ ID NOS: 55, 56 and 57; SEQ ID NOS: 58, 59 and 60; 61, 62 and 63; SEQ ID NOS: 64, 65 and 66; SEQ ID NOS: 67, 68 and 69; SEQ ID NOS: 70, 71 and 72; SEQ ID NOS: 73, 74 and 75; SEQ ID NOS: 76, 77 and 78; SEQ ID NOS: 79, 80 and 81; SEQ ID NOS: 82, 83 and 84; SEQ ID NOS: 85, 86 and 87; SEQ ID NOS: 88, 89 and 90; SEQ ID NOS: 91, 92 and 93; SEQ ID NOS: 94, 95 and 96; And an oligonucleotide primer set of SEQ ID NOS: 97, 98, and 99, which comprises at least one species selected from the group consisting of Korean ginseng, American ginseng, Chinese ginseng, japanese ginseng, Vietnamese ginseng, A set of Kompetitive Allele Specific PCR (KASP) primers. Isolating the genomic DNA from the ginseng sample;
Amplifying the target sequence by performing an amplification reaction using the separated genomic DNA as a template and using the primer set according to claim 2 or 3; And
A method for distinguishing one or more species from a species of ginseng including Korean ginseng, American ginseng, Chinese ginseng, japanese ginseng, Vietnamese ginseng, ginseng fox ginseng, and lesion ginseng, comprising the step of analyzing the product of the amplification step. 5. The method of claim 4, wherein the analysis of the product of the amplification step is performed by restriction enzyme cleavage, fluorescence measurement or phosphorescence measurement. A primer set according to claim 2 or 3; And a reagent for carrying out an amplification reaction, wherein the kit comprises at least one species selected from the group consisting of Korean ginseng, American Samoan, Chinese medicinal herb ginseng, Korean medicinal herb, Vietnamese ginseng, 7. The kit of claim 6, wherein the reagent for performing the amplification reaction comprises DNA polymerase, dNTPs, and a buffer.

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