Application of DNA bar code in identification of authenticity, production place or batch of Dendropanax dentiger product

Abstract

The invention discloses an application of a DNA bar code in identification of authenticity, production places or batches of Dendropanax dentiger products. The DNA bar code is selected from at least one of a gene region polymorphic fragment ycf1 and an intergene region polymorphic fragment ndhFrpl32. Researches find that ycf1 and ndhFrpl32 in chloroplast of the Dendropanax dentiger and related species of the Dendropanax dentiger have various haplotypes, and the Dendropanax dentiger and the related species of the Dendropanax dentiger respectively have at least one specific haplotype; and the haplotypes of ycf1 or ndhFrpl32 owned by the dendropanax dentiger from different producing areas are also different. Therefore, by detecting the haplotype of at least one of ycf1 and ndhFrpl32 in the chloroplast of the Dendropanax dentiger product, the authenticity of the Dendropanax dentiger product and the product can be quickly and accurately identified; and batch anti-counterfeiting of the Dendropanax dentiger product can be realized by adopting the DNA bar code.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biotechnology, and particularly relates to application of DNA barcoding in identifying authenticity, origin or batch of Dendropanax dentiger. BACKGROUND

[0002] According to Chinese Herbal Medicine, Dendropanax dentiger (Harms) Merr. in Dendropanax is used as medicine with roots or branches and leaves, which is sweet and pungent in flavor, and belongs to liver and lung channels. Dendropanax dentiger contains various chemical components of structural types, mainly polyyne, terpenoids, saponins, volatile oil, etc.; and can be used for the treatment of paralysis, migraine, brachial plexus neuritis, rheumatic and rheumatoid arthritis, sprain, furuncle, post-polio syndrome, menstrual disorders, etc., and has pharmacological activities of anti-arrhythmia, anti-cancer, anti-bacterial, anti-rheumatism, anti-atherosclerosis, inhibition of tyrosinase, anti-hepatitis virus, etc. Dendropanax plants are commonly used folk medicines, which are recorded in Guizhou Folk Medicine, Zhejiang Folk Medicine, Guangxi Herbal Selection, Fujian Pharmacopoeia, Jiangxi Herbal Medicine, and National Chinese Herbal Medicine Compilation. With the in-depth research on Panax, Sanqi, Eleutherococcus, American ginseng and other precious tonic and invigorating traditional Chinese medicines, the research on Panax relatives has also attracted great interest.

[0003] Dendropanax dentiger is one of 30 She medicines with higher frequency of use in Zhejiang and Jiangxi, and the quality thereof directly affects the clinical medication effect and patient safety. There are few market products of Dendropanax dentiger, and there is no cultivated product of Dendropanax dentiger at present. The wild Dendropanax dentiger is mainly used in clinical medication, and it is easy to confuse the close relatives when picked by farmers and plant lovers, resulting in differences in clinical medication.

[0004] There are many close relatives of Dendropanax dentiger that are easy to confuse, such as Dendropanax proteus (Champ. ex Benth.) Benth. in Araliaceae Dendropanax, Brassaiopsis fatsioides Harms in Araliaceae Brassaiopsis, Metapanax davidii (Franch.) J. Wen ex Frodin in Araliaceae Metapanax, and other similar plants in Metapanax. It is difficult to distinguish these close relatives from each other by appearance in hospitals and pharmacies at present. Through germplasm resource identification, the purity of Dendropanax dentiger can be ensured, and the confusion and substitution of products can be prevented, so as to ensure the quality of medicinal materials.

[0005] However, research on the identification of *Panax japonicus* germplasm resources in China is relatively scarce, and there are no reports of scholars conducting identification studies on *Panax japonicus* and its common, easily confused closely related species. Some scholars have proposed that the quality control of medicinal materials can be achieved through place of origin identification. However, place of origin identification has always been a major challenge in medicinal material identification. On the one hand, because the morphological and tissue differences of medicinal materials from different places are very small, relying solely on traditional Chinese medicine identification methods—empirical identification and microscopic identification—cannot fundamentally solve the problem of mixed samples in the medicinal material market. On the other hand, because the differences in effective components of medicinal materials from different places are affected by factors such as growth years and sampling time, chemical identification also makes it difficult to delineate the chemical characteristics of the same medicinal material from different places. *Panax japonicus* species are particularly difficult to identify due to the large number of species and the overlap of names and varieties during the sales process. Currently, there is limited research on how to trace the origin of *Panax japonicus* medicinal materials, distinguish different production batches, differentiate mixed products, screen high-quality varieties, and identify growth years.

[0006] Compared with traditional Chinese medicine (TCM) identification methods, DNA barcoding technology offers multiple advantages for identifying medicinal materials, including high efficiency, accuracy, versatility, objectivity, expanded detection range, promotion of standardization and automation, and facilitation of the TCM industry. These advantages make DNA barcoding technology a promising field for TCM identification. Chloroplasts are organelles unique to plant cells, containing double-stranded circular DNA. Plant chloroplast DNA evolves 3-4 times faster than mitochondrial DNA, particularly in the evolution of chloroplast gene sequences. Furthermore, chloroplast DNA is maternally inherited in most angiosperms and flows through seed flow, thus exhibiting a higher degree of geographical structure than nuclear genes, i.e., differentiation between different regions. Therefore, it can be used for the identification and analysis of TCM germplasm resources.

[0007] Currently, there are few reports, both domestically and internationally, of gene polymorphic regions that can be used for the identification of tree ginseng and its easily confused closely related species. Summary of the Invention

[0008] The present invention provides an application of DNA barcoding in identifying the authenticity, origin, or batch of Gynostemma pentaphyllum products, providing a fast and accurate method for identifying the germplasm of Gynostemma pentaphyllum and its closely related species, identifying the origin of Gynostemma pentaphyllum, and identifying different batches of Gynostemma pentaphyllum.

[0009] To achieve the above-mentioned objectives, the technical solution of the present invention is as follows:

[0010] An application of DNA barcoding in identifying the authenticity of tree ginseng products, wherein the DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

[0011] This invention is the first to discover that the ycf1 gene region polymorphic fragment and the ndhF_rpl32 intergenetic region polymorphic fragment in chloroplasts both have at least 17 haplotypes, and that Dendropanax dentiger (Harms) Merr. and many closely related species of Dendropanax dentiger each possess at least one specific haplotype. Therefore, by detecting at least one haplotype of ycf1 and ndhF_rpl32 in the chloroplasts of Dendropanax dentiger products, the authenticity of Dendropanax dentiger products (i.e., whether the Dendropanax dentiger product is Dendropanax dentiger (Harms) Merr.) can be quickly and accurately identified.

[0012] That is, the aforementioned DNA barcode can at least be used to identify Dendropanax dentiger (Harms) Merr. and its closely related species, including at least Dendropanax proteus (Champ. ex Benth.) Benth., Metapanax davidii (Franch.) J. Wen ex Frodin., and Brassaiopsis fatsioides Harms.

[0013] This invention has revealed that not only do *Gnaphalium affine* and its closely related species possess different haplotypes of ycf1 or ndhF_rpl32, but haplotypes of ycf1 or ndhF_rpl32 also differ among *Gnaphalium affine* species from different origins. Therefore, this invention also provides the application of the aforementioned DNA barcodes in identifying the origin of *Gnaphalium affine* products.

[0014] Preferably, the aforementioned DNA barcode is used to identify the origin of Dendropanax dentiger (Harms) Merr., which includes at least Chenzhou City in Hunan Province, Yichun City in Jiangxi Province, Shaowu City in Fujian Province, Ji'an City in Jiangxi Province, Jiujiang City in Jiangxi Province, and Laibin City in Guangxi Zhuang Autonomous Region.

[0015] Since the DNA barcode can effectively identify the authenticity and origin of ginseng products, it can also be used to identify different batches of ginseng products when it is necessary to identify several batches of ginseng products to determine whether the quality of each batch is stable, so as to achieve batch anti-counterfeiting of ginseng products.

[0016] That is, the present invention also provides the application of the above-mentioned DNA barcode in identifying batches of tree ginseng products, characterized in that the DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

[0017] Similarly, the present invention also provides an application of a DNA barcode detection kit in identifying the authenticity, origin, or batch of tree ginseng products, wherein the DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

[0018] Preferably, the detection kit contains specific amplification primers for at least one of the gene region polymorphism fragment ycf1 and the intergene region polymorphism fragment ndhF_rpl32.

[0019] In this invention, either the gene region polymorphic fragment ycf1 or the intergene region polymorphic fragment ndhF_rpl32 can be used as a DNA barcode, or ycf1 and ndhF_rpl32 can be used together as a DNA barcode. The results are more accurate when analyzed together.

[0020] Compared with the prior art, the technical effects of the present invention are reflected in:

[0021] (1) This invention is the first to discover that the polymorphic fragment ycf1 in the gene region and the polymorphic fragment ndhF_rpl32 in the intergenetic region of chloroplasts both have multiple haplotypes, and that Dendropanax dentiger (Harms) Merr. and many closely related species of Dendropanax dentiger each possess at least one specific haplotype. Moreover, the haplotypes of ycf1 or ndhF_rpl32 possessed by Dendropanax dentiger from different origins also differ. Therefore, by detecting at least one haplotype of ycf1 and ndhF_rpl32 in the chloroplasts of Dendropanax dentiger products, the authenticity of Dendropanax dentiger products (i.e., whether the Dendropanax dentiger product is Dendropanax dentiger (Harms) Merr.) and the product itself can be quickly and accurately identified.

[0022] (2) Since the DNA barcode of the present invention can effectively identify the authenticity and origin of ginseng products, when it is necessary to identify several batches of ginseng products to determine whether the quality of each batch is stable, the above-mentioned DNA barcode can also be used to identify different batches of ginseng products in order to achieve batch anti-counterfeiting of ginseng products.

[0023] (3) In this invention, one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32 can be used as a DNA barcode, or ycf1 and ndhF_rpl32 can be combined as a DNA barcode. The results are more accurate when combined analysis. Attached Figure Description

[0024] Figure 1 Pi diagram of chloroplast genome nucleotide diversity for *Gynostemma pentaphyllum* and its easily confused close relatives;

[0025] Figure 2mVISTA diagram of chloroplast genome sequence homology analysis of *Panax quinquefolius* and its easily confused closely related species;

[0026] Figure 3 Phylogenetic tree constructed from haplotype sequences of ycf1 and ndhF_rpl32 after joint analysis;

[0027] Figure 4 Genetic distance constructed from haplotype sequences after joint analysis of ycf1 and ndhF_rpl32;

[0028] Figure 5 The ycf1 sequencing peak diagrams for haplotypes AHap1 (HNZJJ) and AHap9 (GXHZ);

[0029] Figure 6 The ndhF_rpl32 sequencing peak diagrams are for haplotypes BHap1 (HNZJJ) and BHap2 (FJNP). Detailed Implementation

[0030] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0031] Example 1: Screening of polymorphic regions in the chloroplast genome of tree ginseng

[0032] 1. Results of chloroplast genome gene region (interregion) alignment

[0033] First, based on the chloroplast genome sequence NC026546 from the NCBI database, we collected samples from four *Gnaphalium affine* samples from Ningbo (Zhejiang), Sanming (Fujian), Ji'an (Jiangxi), and Zhangjiajie (Hunan), one *Gnaphalium affine* sample from Wuzhou (Guangxi), one *Tetracentron sinense* sample from Bijie (Guizhou), and one *Tetracentron sinense* sample from Laibin (Guangxi). Chloroplast genomes were extracted and sequenced. Nucleotide diversity (Pi) of the chloroplast genome sequences of these seven *Gnaphalium affine* samples and their easily confused relatives was analyzed using DNA SP6. The results are as follows: Figure 1 As shown.

[0034] Figure 1 The results showed that the Pi value ranged from 0.00000 to 0.02833, and genes such as trnK-rps16, atpF, trnC-petN, trnT-psbD, trnT-trnL, psbE-petL, petD, ndhF-rpl32, and ycf1 could serve as potential origin-specific DNA barcodes for *Gnaphalium affine* and its easily confused closely related species.

[0035] To detect the overall variation of intraspecific and extraspecific chloroplast genomes in *Gynostemma pentaphyllum*, using the annotated *Gynostemma pentaphyllum* chloroplast genome sequence (NC_026546) as a reference, the mVISTA online software was used to perform a global comparison of the chloroplast genomes of seven *Gynostemma pentaphyllum* species and their easily confused close relatives. The results are as follows: Figure 2 As shown.

[0036] The chloroplast genome of *Gnaphalium affine* obtained by sequencing was globally compared with the whole chloroplast genome sequence of *Gnaphalium affine* in GenBank using the mVISTA online software. Genes such as trnQ-UUG, trnG-UCC, rpoC2, psbE, rps19, ndhF-rpl32, and ycf1 can be used as potential origin-specific DNA barcodes for *Gnaphalium affine* and its easily confused closely related species.

[0037] 2. Screening for polymorphic regions of chloroplast genes and intergenetic regions in tree ginseng and its easily confused closely related species.

[0038] For an efficient barcode sequence to be effective, in addition to meeting the requirement of sufficient variability to distinguish different species, it also needs to meet the experimental requirements of PCR amplification and PCR product sequencing. Therefore, sequence length is also a selection factor. Experiments have shown that gene fragments of *Gnaphalium affine* and its easily confused closely related species with sequence lengths between 600 and 1600 bp have higher amplification efficiency and better sequencing results.

[0039] To enrich as many haplotypes as possible to distinguish between different populations of *Gynostemma pentaphyllum* and its easily confused closely related species, we focused on gene polymorphism abundance, combined with nucleotide diversity and mVISTA results, and screened out the ycf1 and ndhF_rpl32 gene fragments as DNA barcodes for the specific identification of the authentic core germplasm of *Gynostemma pentaphyllum* and its easily confused closely related species.

[0040] Example 2: Identification of different authentic core germplasm of Gynostemma pentaphyllum and identification of Gynostemma pentaphyllum and its easily confused closely related species using DNA barcoding.

[0041] 1. Haplotype sequencing of ycf1 and ndhF_rpl32 gene fragments

[0042] Multiple samples of Gynostemma pentaphyllum from different regions across the country were collected (see Table 5) as well as multiple samples of Gynostemma pentaphyllum from easily confused closely related species (see Table 6). (Each numbered sample contained at least 10 plants. During PCR amplification and sequencing, each plant was tested separately, and the sequencing results of different plants could be mutually verified.)

[0043] DNA was extracted from each sample, and the ycf1 and ndhF_rpl32 fragments were amplified by PCR using the two pairs of specific primers shown in Table 1 below. After gel recovery, the fragments were sequenced.

[0044] Table 1. Specific primers for ycf1 and ndhF_rpl32 fragments

[0045] Primer name Primer sequence (5'-3') ycfl-F CTATTATCAATGAAGGTA (SEQ ID No. 1) ycfl-R GAATCAGGATCGAGTTTA (SEQ ID No. 2) ndhF_rpl32-F TCAAATGAGATGACCAAC (SEQ ID No. 3) ndhF_rpl32-R AGAATCCAAAAAAAGACC (SEQ ID No. 4)

[0046] 2. Using ycf1 to analyze the haplotype sequence characteristics of *Panax notoginseng* from different regions and its easily confused closely related species.

[0047] Analysis of the sequencing results revealed that the ycf1 fragment has a total of 17 haplotypes. The sequences of various ycf1 gene haplotypes are shown in SEQ ID NO.5-21, namely ycf1AHap1-AHap17 (the primers in Table 1 are designed based on ycf1AHap1, and the same applies to ndhF_rpl32).

[0048] The characteristics of each ycf1 haplotype sequence were analyzed, and the results are shown in Table 2.

[0049] Table 2. Results of sequence feature analysis for each ycf1 haplotype.

[0050]

[0051]

[0052] Based on the analysis results, the 17 haplotypes of the ycf1 gene can be divided into two main categories:

[0053] The first type of mutation sites (mutations relative to AHap1) are mainly distributed between 63 and 494 bp, and there are 10 main haplotypes (AHap1, AHap2, AHap3, AHap4, AHap6, AHap7, AHap9, AHap10, AHap11, AHap17). The specific bases at positions 63, 104, 114, 116, 173, 202, 221, 265, 285, 290, 306, 346, 352, 353, 417, and 494 are T, A, T, A, C, T, C, T, G, G, A, C, A, T, G, T, respectively; bases at positions 139-144... Deletions exist; an insertion sequence T exists between positions 127-128, and an insertion sequence TTTTTC exists between positions 152-153; the specific base at position 444 of haplotypes AHap3 and AHap4 is C; the specific base at position 132 of haplotype AHap3 is C; the specific base at position 258 of haplotype AHap7 is A; and the specific bases at positions 152, 182, 248, 259, 272, 284, 421, 437, 439, 442, 446, 451, 463, 481, and 488 of haplotype AHap17 are G, C, T, T, A, G, A, A, A, G, C, C, G, A, T, G, respectively.

[0054] The second type of mutation sites (mutations relative to AHap1) are distributed in the range of 495–759 bp, mainly consisting of 7 haplotypes (AHap5, AHap8, AHap12, AHap13, AHap14, AHap15, and AHap16). The specific bases at positions 502, 545, 570, and 668 are A, C, G, and T, respectively. There are deletions at positions 699, 736, 738, and 739, and an insertion sequence T exists between positions 736 and 737. The specific bases at positions 736, 737, and 740 of haplotypes AHap8, AHap12, and AHap14 are C, T, and G, respectively.

[0055] Association analysis of the ycf1 gene haplotypes with various *Panax japonicus* and its easily confused closely related samples revealed (see Tables 5 and 6) that some *Panax japonicus* samples and closely related samples from certain producing areas possess their own unique haplotypes. Specifically, Rucheng County, Chenzhou City, Hunan Province possesses the unique haplotype AHap5 (i.e., haplotype AHap5 was only found in *Panax japonicus* samples from Chenzhou City, Hunan Province; it was not found in *Panax japonicus* samples from other producing areas or closely related species, the same below); Laozhushan, Yuanzhou District, Yichun City, Jiangxi Province possesses the unique haplotype AHap7; and Nanping City, Fujian Province possesses the unique haplotype AHap8. *Panax variegata* possesses unique haplotypes AHap9, AHa10, AHap11, AHap12, AHa13, AHap14, AHap15, and AHap16. *Panax variegata* possesses the unique haplotype AHap17.

[0056] The above demonstrates that the ycf1 gene can be used for intraspecific identification of ginseng (for example, if the ycf1 gene of a ginseng sample is detected to be of the AHap5 haplotype, then the origin of the ginseng sample can be confirmed as Binzhou City, Hunan Province) as well as for interspecific identification of ginseng and its easily confused close relatives.

[0057] 3. Analyze the haplotype sequence characteristics of *Panax notoginseng* and its easily confused closely related species from different regions using ndhF_rpl32.

[0058] Analysis of the sequencing results showed that the ndhF_rpl32 fragment has 17 haplotypes ( Figure 2 The sequences of various ndhF_rpl32 gene haplotypes, namely ndhF_rpl32BHap1-BHap22, are shown in SEQ ID NO.:22-43. The sequence characteristics of the ndhF_rpl32 haplotypes were analyzed, and the results are shown in Table 3.

[0059] Table 3. Results of sequence feature analysis for each ndhF_rpl32 haplotype.

[0060]

[0061]

[0062] Based on the analysis results, the 17 haplotypes of the ndhF_rpl32 fragment can be divided into two main categories:

[0063] The first type of mutation sites are mainly distributed between 208 and 591 bp, and there are 13 haplotypes (BHap1, BHap2, BHap3, BHap5, BHap6, BHap7, BHap8, BHap10, BHap11, BHap12, BHap13, BHap15, BHap16, BHap17). Among them, the specific bases at positions 232, 256, 279, 320, 321, 334, 387, 408, 442, 468, 469, 471, 488, 523, 561, 585, and 591 are A, T, T, respectively. G, G, T, G, C, T, A, C, A, T, T, G, A, A; a deletion exists at position 321; a deletion exists at positions 473-478; and an insertion sequence T exists between positions 208-209. Haplotypes BHap2, BHap7, BHap11, BHap12, and BHap13 have specific bases A, G, G, C, T, T, G, A at positions 232, 321, 387, 408, 442, 523, 561, and 585. Haplotype BHap7 has a specific base T at position 529, and haplotype BHap13 has a specific base T at position 547.

[0064] The second type of mutation sites are mainly distributed in the 656-777bp range, with four main haplotypes (BHap4, BHap6, BHap9, and BHap14). The specific bases at positions 686, 708, and 777 are T, G, and G, respectively. There are deletions at positions 731-771, and an insertion sequence AATTACTCATTCAAA at positions 656-657. The specific bases at positions 708 and 768 of the exophytic haplotype BHap14 are G and G, respectively.

[0065] Association analysis of haplotypes of the ndhF_rpl32 gene with various *Panax notoginseng* and its easily confused closely related species revealed (Tables 5 and 6) that *Panax notoginseng* samples and *Panax notoginseng* closely related species from certain producing areas possess their own unique haplotypes. For example, Wan'an County, Ji'an City, Jiangxi Province, possesses the unique haplotype BHap6; Rucheng County, Chenzhou City, Hunan Province, possesses the unique haplotypes BHap4 and BHap9; and Xiushui County, Jiujiang City, Jiangxi Province, possesses a unique haplotype...

[0066] BHap8. *Gynostemma pentaphyllum* possesses exclusive haplotypes BHap10, BHap11, BHap12, and BHap13. *Gynostemma discoidum* possesses an exclusive haplotype BHap14. *Gynostemma heterophyllum* possesses exclusive haplotypes BHap15, BHap16, and BHap17. This demonstrates that the ndhF_rpl32 gene can be used for intraspecific identification of origin in *Gynostemma pentaphyllum* and for interspecific identification of *Gynostemma pentaphyllum* and its easily confused closely related species.

[0067] 4. Perform joint analysis using ycf1 and ndhF_rpl32 fragments.

[0068] Joint analysis of the ycf1 and ndhF_rpl32 fragments yielded 30 haplotypes (see Table 4), namely Hap1-Hap30.

[0069] Table 4 shows the 30 haplotypes identified through joint analysis.

[0070]

[0071]

[0072] Joint analysis of the ycf1 and ndhF_rpl32 genes revealed (see Tables 5 and 6) that each production area possesses its own unique haplotype, resulting in more accurate results and the ability to identify more production areas. For example, Rucheng County in Chenzhou City, Hunan Province, has unique haplotypes Hap10 and Hap13; Laozhushan in Yuanzhou District, Yichun City, Jiangxi Province, has unique haplotypes Hap14 and Hap15; Shaowu City in Nanping City, Fujian Province, has unique haplotypes Hap3 and Hap16; Xiushui County in Jiujiang City, Jiangxi Province, has unique haplotype Hap9; Jinxiu Yao Autonomous County in Laibin City, Guangxi Zhuang Autonomous Region, has unique haplotype Hap11; Wan'an County in Ji'an City, Jiangxi Province, has unique haplotype Hap12; and Wuning County in Jiujiang City, Jiangxi Province, has unique haplotype Hap6. *Gynostemma pentaphyllum* possesses exclusive haplotypes Hap17, Hap18, Hap19, Hap20, Hap21, Hap22, Hap23, Hap24, Hap25, and Hap26; *Gynostemma variegata* possesses the exclusive haplotype Hap27; and *Gynostemma heterophyllum* possesses exclusive haplotypes Hap28, Hap29, and Hap30. This indicates that combined analysis of the ycf1 and ndhF_rpl32 genes can be used for intraspecific identification of *Gynostemma pentaphyllum* and interspecific identification of *Gynostemma pentaphyllum* with easily confused closely related species.

[0073] Table 5. Collection table of *Panax ginseng* medicinal materials from different regions and corresponding ycf1 fragment haplotypes.

[0074]

[0075]

[0076] Table 6. Collection of Samples of Easily Confused Closely Related Species of Ginseng.

[0077]

[0078] 4. Using ycf1 and ndhF_rpl32 combined analysis, we analyzed the phylogenetic evolution and genetic distances of *Gnaphalium affine* from different regions and its easily confused closely related species.

[0079] Phylogenetic trees and genetic distances were constructed using MEGA 11 software for haplotype sequences of *Panax japonicus* and its easily confused closely related species. Figure 3 As can be seen, the phylogenetic tree constructed from haplotype sequences can be divided into two major branches, similar to the sequencing results analysis above; Hap12 and Hap20 form one major branch with 100% support; Hap1-Hap11, Hap13-Hap19, and Hap21-Hap30 are distributed on the other branch with 59% support.

[0080] from Figure 4 It can be seen that the genetic distances of haplotype sequences after joint analysis of ycf1 and ndhF_rpl32 in different species of *Gnaphalium affine* and its easily confused closely related species range from 0 to 0.34936, with an intraspecific average genetic distance of 0.02166. The genetic distance between haplotypes Hap28 and Hap29 is the smallest, at 0.00573, while the genetic distance between Hap6 and Hap20 is the largest, at 0.34936. The genetic distance analysis results are consistent with the phylogenetic tree results.

[0081] Example 3: Using DNA barcoding for batch anti-counterfeiting of tree ginseng products

[0082] Taking the selected sequences ycf1 and ndhF_rpl32 as examples, a batch anti-counterfeiting study was conducted on *Gnaphalium affine* and its easily confused closely related species.

[0083] 3.1 Using the ycf1 intergenic region sequence for batch anti-counterfeiting verification of *Gynostemma pentaphyllum* and its easily confused closely related species.

[0084] The experiment used chloroplast gene region fragments as the main means to establish batch barcodes for Chinese medicinal materials. The batch barcodes used in the experiment included, but were not limited to, chloroplast gene region fragments such as ycf1 and ndhF_rpl32.

[0085] Leaves from two different haplotypes, HNZJJ haplotype AHap1 in Table 1 and GXHZ haplotype AHap9 in Table 2, along with their easily confused closely related species, were selected and mixed for DNA extraction. The ycf1 fragment was amplified using PCR, recovered from the gel, and sequenced. The sequencing peak diagram is shown below. Figure 5 As shown.

[0086] Depend on Figure 5The following ycf1 fragment mutation sites are clearly visible: at 202bp, AHap1 is G and AHap9 is T, with a GT peak structure in the sequencing peak spectrum; at 221bp, AHap1 is T and AHap9 is C, with a TC peak structure in the sequencing peak spectrum; at 285bp, AHap1 is A and AHap9 is G, with an AG peak structure in the sequencing peak spectrum; at 290bp, AHap1 is A and AHap9 is G, with an AG peak structure in the sequencing peak spectrum; at 346bp, AHap1 is A and AHap9 is C, with an AC peak structure in the sequencing peak spectrum; and at 495bp, AHap1 is C and AHap9 is T, with a CT peak structure in the sequencing peak spectrum.

[0087] Sequencing results show that the ycf1 fragment peak diagrams of *Gnaphalium affine* and its easily confused closely related species can identify their haplotypes, with obvious variation sites and overlapping peak structures. This fragment can be used as a DNA barcode for batch anti-counterfeiting.

[0088] 3.2 Batch anti-counterfeiting verification of *Gynostemma pentaphyllum* and its easily confused closely related species was performed using the intergenic region sequence of ndhF_rpl32.

[0089] Leaves from two different haplotypes of *Pterocarya stenoptera* (HNZJJ haplotype BHap1 and FJNP haplotype BHap2, as shown in Table 1) were selected and mixed for DNA extraction. The ndhF_rpl32 fragment was amplified using a PCR instrument, recovered from the gel, and then sequenced. The sequencing peak diagram is shown below. Figure 6 As shown.

[0090] Depend on Figure 6 The following mutation sites are clearly visible in the ndhF_rpl32 fragment: at 523bp, BHap1 is G and BHap2 is T, and the sequencing peak pattern at this site shows a GT nested peak structure; at 561bp, BHap1 is T and BHap2 is G, and the sequencing peak pattern at this site shows a TG nested peak structure; at 585bp, BHap1 is C and BHap2 is A, and the sequencing peak pattern at this site shows a CA nested peak structure; at 686bp, BHap1 is C and BHap2 is T, and the sequencing peak pattern at this site shows a CT nested peak structure; at 708bp, BHap1 is T and BHap3 is G, and the sequencing peak pattern at this site shows a TG nested peak structure.

[0091] Sequencing results show that the peak diagrams of the ndhF_rpl32 fragment in each batch of *Panax quinquefolius* and its easily confused closely related species can identify the haplotype species, with obvious variation sites and overlapping peak structures. This fragment can be used as a DNA barcode for batch anti-counterfeiting.

[0092] The base sequences of each haplotype of ycf1 and each haplotype of ndhF_rpl32 involved in this invention are as follows:

[0093] 1.ycf1 haplotype base sequence

[0094] >AHap1

[0095] AAAAATTGGAGTTCTATTATCCAAGAAGGTAATAATATATTTTCTAAGCTTCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACGAGATCCCATATTTCCGGGATTTCTATCCGCTTTTTCCTTTATTTTGTTCTCCTCTTGTTTCTCTCTTCTTTTTGTCTGTTCTTCTGTATAATCT TTTAATTCTGTCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAAGGGGATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCAAAATAGGAGTTTTACGCCTTTGAGCACGCATAGAACCTTTGATTATGTCTCGACGAAAATCCG ATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTACATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAACTCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTT ACTGATTTCTTTTATTCCAAAAGATTTTTGTTTTATTTTGTGACCATTAGGTCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTCAAATTTAAACTCATTCCCTGATTCAAATTTTTTTTTTTTTTTGTTTAGG

[0096] >AHap2

[0097] TTTTCTCTTAATATCAAAAAAAGGTTATATAATATATTTTCTAAGTCTCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGGGTTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGACGA AAATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTAAAATTTAAACCGTCCCCGGATTAACAAA

[0098] >AHap3

[0099] TCTTTTTTCTTTCTCACCAAAAATGGTAAAATAATAATATTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGAAAATGGAACTAAATCCCATATTTCCGGCATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGTAGATATCAAAAGAAAAAAAGGGGGATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAAGAGTTTTACGCCTTTGAGCACGCATAGAACCTTTGATTATGTCTCGACGAAA ATCCGATTCTTGGGAATATTCTATCGTATATACCAGGTCCATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAAGTCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTGTTTTATTTTGTGACCATTAGGACCAGTTATAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTAAAATTTAACACCCCCCCTGCTTATAGCAATAAATG

[0100] >AHap4

[0101] TTTTCTTTTTCTCACCCAATAGGTAATATAATATATTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGAAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGTAGATATCAAAAGAAAAAAAGGGGGATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAAGAGTTTTACGCCTTTGAGCACGCATAGAACCTTTGATTATGTCTCGACG AAAATCCGATTCTTGGGAATATTCTATCGTATATACCAGGTCCATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAAGTCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTGTTTTATTTTGTGACCATTAGGACCAGTTATAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTAAAATTTAACCGAGCCCCGGTTCACA

[0102] >AHap5

[0103] TTTTCTTTTTATACCCCAGAAGGTTATATAATATATTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGAAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGTAGATATCAAAAGAAAAAAAGGGGGATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAAGAGTTTTACGCCTTTGAGCACGCATAGAACCTTTGATTATGTCTCGACGAA AATCCGATTCTTGGGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAAGTCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTGTTTTATTTTGTGACCATTAGGACCAGTTATAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTTCTGAAAATAAAGAAAGGCCCTTAAAATTTAAACTCTTATCCCTGAGATCACA

[0104] >AHap6

[0105] CCTTTTTTTTCCAAAAGAGGTAATATAATATATTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGAAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGTAGATATCAAAAGAAAAAAAGGGGGATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAAGAGTTTTACGCCTTTGAGCACGCATAGAACCTTTGATTATGTCTCGACGAAAAT CCGATTCTTGGGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAAGTCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTGTTTTATTTTGTGACCATTAGGACCAGTTATAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTAAAATTTAAACTCGAACCCTGATTCAAATCCAA

[0106] >AHap7

[0107] TTCCTTTTTCCAATAAGGTAATATAATATATTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGAAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATAAGTTCGTAGATATCAAAAGAAAAAAAGGGGGATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAAGAGTTTTACGCCTTTGAGCACGCATAGAACCTTTGATTATGTCTCGACGAAA ATCCGATTCTTGGGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAAGTCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGACCAGTTATAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTAAAATTTAAACCGACCCCTGGATTCAA

[0108] >AHap8

[0109] TTTCTCTATATTACACACAATAGGTATATAATATTTTTTTCTAAGTCTCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGGGTTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGACGA AAATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTCTGAAATAAAGAAAGGCCCTTAATTTAACATACCCTCGTCGTAGTATCACA

[0110] >AHap9

[0111] CTTTTTTTTTGCCAAAGGGTAATATAATAATATTTTCTAAGTCTCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGGGTTTTTTTCTTCTGTACAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGACGAAAA TCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGGCCCTTAAAATTTAAATCGTCCCCGGGTGAATCACAA

[0112] >AHap10

[0113] TCTCTATTATCTACAAAAGAGGTAATATAATATATTTTCTAAGTCTCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGGGTTTTTTTCTTCTGTACAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGACGAAA ATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTCTGAAAATAAAGAAAGGCCCTTAAATTATCGACTCTGCTGCTCGCGTCGTCTGC

[0114] >AHap11

[0115] TTTTTTTTTTTTATCAAAAGAGTATATAATATATTTCTAAGTCTCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGGGTTTTTTTCTTCTGTACAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGACGAA AATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTCTGAAAATAAAGAAAGGCCCTAAATTATGATACTCTCGCTAGTATCATACA

[0116] >AHap12

[0117] ACCCATCCTATTATCAAAGAAGGTAATATAATATATTTTCTAAGTCTCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGGGTTTTTTTCTTCTGTACAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGA CGAAAATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAGTTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTTGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTCTGAAATAAAGAAAGGCCCTTAAAATTTAAACTCGTTCCCTGGATTCA

[0118] >AHap13

[0119] CCCCCTCTCTTTCCCCCACAAGGGTAATATAATTATATTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTCTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGAATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGACG AAAATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAATTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTCGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTAAAATTTAAACCGACCCCCGCCGAGTATCACAAA

[0120] >AHap14

[0121] TTTCTCTCTTATACTCCACCATAGTATATATATTTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTCTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGAATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCG ACGAAAATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAATTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTCGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTCTGAAATAAAGAAAGGCCCTTAAATTTACTCGTTCCCCGTGTATACA

[0122] >AHap15

[0123] TTCTATTATCCAAAGAGGTAATATAATATATTTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTCCGGGATTTCTATCCGCGCTTTTTCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTCTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGAATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTAT GTCTCGACGAAAATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAATTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTCGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTTCTGAAAATAAAGAAAGGCCCTTAAATTTAAT

[0124] >AHap16

[0125] TTTTCTTTTTTCCACACATAGGTAATATAATATATTTCTAAGTATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGGAAATGGAACTAAATCCCATATTTTCCGGGATTTCTATCCGCGCTTTTTCTTTTTCCTTTATTTTGTTCTCCTCTTCTTTCTCTCTTCTTTTTGTCTGTTCTTCTTTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAAGAAGTTTCATCAGTTCGGAGATATCAAAAGAAAAAAAGGGGAATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAATAGTTTTACGCCTTTGAGCAGCATAGAACCTTTGATTATGTCTCGACGA AAATCCGATTCTTGCGAATATTCTATCGTATATACCAGGTCTATTTGATCAAAATTATCAGCATTCCATTTGTTAGGAGTAAAAATTACTATATCGTCAATTTTTCTTGAACGAATCTGATGTCCTACCGGTAAGCCTTCTCGAACTACGCCCGACTGTTGTTCCAACGCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAGGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTAATTTACCGTACTCTCGCTGCTGATATCACCA

[0126] >AHap17

[0127] TCTTAATTAAACACAATAGGTAATATAATATATTTTCTAAGCATCGATTGAATTAGTAATATAAAACTTCTTGTTGTTTGCGAAAATGGAACTAAATCCCATATTTCCGGGATTTCTGCTTTTTGCTTTATTTTGTTCTCCTCTTGTTTCTCTCCTCTTTTTGTCTGTTCTTCTGTATAATCTTTTAATTCTGCCCCTTTACCCATCCAATTTCTAAAAATAAGTTTCATCTGTTCGGAGATATAAAAAGAAAAAAGGGGGGATTTTTTTCTTCTGTCCAAAAAAAGTGGGGAATGCGCATTTGCTTGAAACAGTTTCCAAATAAGAGTTTTACGCCTTTGAGCACGCATAGAACCTTTGATTATGTCTCGACGAAAATCCGATTCTTGCGAAAATTCTATCGTATATAACGGGCCTACTTGAGCAAAATTATCAACATTCCATTTGTTAGGATTAAAAAGTACTATATCGTCAATTTTTCTTGAACGAATCTGATGTCCTACCGGCAAGCCTTCTTGAACTACGCCCGTCTGTTGTTCCAACTCGGTGATAAGTTTGTATGACCATCGAGGGACTTTTTTACTGATTTCTTTTATTCCAAAAGATTTTTTTTTTATTTTGTGACCATTAAGGCCAGTTAGAATTGCATTTAATAAAAATTTTAGAAGTTTTGCTTCATTTTCTGAACCACTTCTTCCTTGTTCTTTTTCTGAAAATAAAGAAAGGCCCTTCAAATTTAAATCAACCCCCTGATTCAAA

[0128] 2.ndhF_rpl32 haplotype base sequence

[0129] >BHap1

[0130] TTTTAAAAACAAAATCCAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAATAACTTTTAGTTAGTTTGTTCAAAATCATTAACTAGTTCATTATGGAATTGATTTATTATTTTCCATTTCAATAAAAAAAACATTTATATTATAGATTATAGAAAACGCTATAATATCTGA CATTTTATATAAAATGAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGTTAAAATAAAATTAATAAAAAAATCACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTCCTCGAGTTTGACCGGTTAGTATAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATTAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTCTTTTTTTTTTGGGATTTCTTA

[0131] >BHap2

[0132] TTTCAAAAGAGGGGACCAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAAATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGAC ATTTTATATAAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTCTTTTGGGGGATTCATCTA

[0133] >BHap3

[0134] TTTTTAAAGCGGAAAAACAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATTAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAGGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGACATT TTATATAAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTTTATATGGTGATGTACATCATACAC

[0135] >BHap4

[0136] TTTTTTAAAAAAAAGGAAAACCAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATTAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAGGAGTACTTGATTTTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAAACATTTATAGATTATATAAAACGCTATAATATCTGACATTTTATATA AAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAAATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAAAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAAAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACGGAGATAAAAAGGAAGGTCTTTTTTTGTGAGATATCTATTA

[0137] >BHap5

[0138] TTTTAAAAAACAAAATCAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATTAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAGGAGTACTTGATTTTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGACAT TTTATATAAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTCTTTTTTTGTGATCCCATAAA

[0139] >BHap6

[0140] TTCCAAAATGAGGAGATCCAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATTAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAGGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGACATTTTATAT AAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAAAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTCTTTTTGGTGGAATTCTT

[0141] >BHap7

[0142] TTCCAATTATATAGTGAGAGTAAAAAAACTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAAATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGGAGTACTTGATTNTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTG ACATTTTATATAAAATTAACTTTATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTTTTATGGGGTAGAATCATCTA

[0143] >BHap8

[0144] TTTTCATAAAAAAAAAGAGAAAACAACTGTTCGTAAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATTAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAGGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGAC ATTTTATATAAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGTCCTTTGTGTGTGTATATCATCAAAA

[0145] >BHap9

[0146] AGACTGATCTTAGTTATTACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATTAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAGGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAAACATTTATAGATTATATAAAACGCTATAATATCTGACATTTTATATA AAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAAAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATGACAGGT

[0147] >BHap10

[0148] TTTTTAAAAAAAAAAAACAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAAATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGACA TTTTATATAAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTCATTGGTGGTGAATCTCACTA

[0149] >BHap11

[0150] CCAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAAATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGGAGTACTTGATTNTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGACA TTTTATATAAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTCTTT

[0151] >BHap12

[0152] GTCAACGCGATACTTAGTTATTAACTAAAATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTG ACATTTTATATAAAATTAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAGAACGAAGAAAATGACAGAAGTG

[0153] >BHap13

[0154] TTTTAAAAAGGGGGATACAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAAATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGGAGTACTTGATTTTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAAGAACTTTTAGTTAGTTTGTTCCAAATCATTAACTAGTTCATTATGGAATTGATTTTTTATTTTCCATTTCAATAAAAAAAAAACATTTATAGATTATAGAAAACGCTATAATATCTGAC ATTTTATATAAAATTAACTTGATTTTCTATTTATCGATTTTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATAACTAAAATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTTCTCGAGTTTGACCGGTTAGTAAAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATGAAATGTAAAACGAAAAAAATAGACAGAGATAAAAAGGAAGGCTTTTTTGGGAATTCAAAAAA

[0155] >BHap14

[0156] AACAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTCCTCGAGTTTGACCGGTTAGTAGAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCTTTATAAATTAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGTCTTTTTTTTTGGAATTCTAAA

[0157] >BHap15

[0158] AAGTCAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATGAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAATAACTTTTAGTTAGTTTGTTCAAAATCATTAACTAGTTCATTATGGAATTGATTTATTATTTTCCATTTCAATAAAAAAAACATTTATATTATAGATTATAGAAAACGCTATAATATC TGACATTTTATATAAAATGAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAAATTAATAAAAAAATCACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTCCTCGAGTTTGACCGGTTAGTATAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATTAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGCTT

[0159] >BHap16

[0160] ACAACCTGTTCGTAAAAAGCGAATACTTAGTTATTAACTAAGATTTTTATGAAGATACCGAAAATTAAAATTTCAATTATTATTACATGAATTTATATTCATAGAGTAAGGATAAAGAATTACACTAAAAGAGTACTTGATTCTGATATGAATCATAGGATTAACTAAGATCAAAAACTTGTCCTAATAAAATAATAACTTTTAGTTAGTTTGTTCAAAATCATTAACTAGTTCATTATGGAATTGATTTATTATTTTCCATTTCAATAAAAAAAACATTTATATTATAGATTATAGAAAACGCTATAATATCTGAC ATTTTATATAAAATGAACTTGATTTTCTATTTATCGATATTTATCGAAAGGGGTAAAATAAATTAATAAAAAAATCACTAAGATCTCTTTTATCAAACCACGTATCCTTTAACAGATTAATAACTTTAATTACTAGTCTCATTCAAATTTTAAAATGGAATTTAGGAGTATTCTTTCCTCGAGTTTGACCGGTTAGTATAAAAAGATTAAAGTTTTCATTAGGCAATGGGTTCTTAAACCCTTTGGTGTATTTTATTCCTTATAAATTAAATGTAGAACGAAGAAAATAGACAGAGATAAAAAGGAAGGCTTTTTTT

[0161] >BHap17

[0162] 。

Claims

1. An application of DNA barcoding in identifying the authenticity of ginseng products, characterized in that, The DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

2. The application as described in claim 1, characterized in that, This is used to identify Dendropanax dentiger (Harms) Merr. and its closely related species, which include at least Dendropanax proteus (Champ. ex Benth.) Benth., Metapanax davidii (Franch.) J. Wen ex Frodin., and Brassaiopsis fatsioides Harms.

3. An application of DNA barcoding in identifying the origin of ginseng products, characterized in that, The DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

4. The application as described in claim 3, characterized in that, This is used to identify the origin of Dendropanax dentiger (Harms) Merr., which includes at least Chenzhou City in Hunan Province, Yichun City in Jiangxi Province, Shaowu City in Fujian Province, Ji'an City in Jiangxi Province, Jiujiang City in Jiangxi Province, and Laibin City in Guangxi Zhuang Autonomous Region.

5. The application of DNA barcoding in batch identification of ginseng products, characterized in that, The DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

6. The application of a DNA barcode detection kit in identifying the authenticity of tree ginseng products, characterized in that... The DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

7. The application of a DNA barcoding detection kit in identifying the origin of ginseng products, characterized in that, The DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

8. The application of a DNA barcode detection kit in batch identification of tree ginseng products, characterized in that, The DNA barcode is selected from at least one of the gene region polymorphic fragment ycf1 and the intergene region polymorphic fragment ndhF_rpl32.

9. The application as described in any one of claims 6-8, characterized in that, The detection kit contains specific amplification primers for at least one of the gene region polymorphism fragment ycf1 and the intergene region polymorphism fragment ndhF_rpl32.

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