CN1982469A - Appraisement of Tangshen variety - Google Patents

Abstract

Tangshen 5S rDNA sequence and discrimination of various Tangshen, pseudo-campanumoea javanica and radix platycodi thereby are disclosed.

Description

Identify the method for Radix Codonopsis kind

Technical field:

The present invention relates to molecular biology, genomics, biotechnology and identification and assessment of Chinese medicines field.

Background of invention:

According to Chinese Pharmacopoeia (version in 2000) Radix Codonopsis is campanulaceae plant Radix Codonopsis Codonopsispilosula (Franch.) Nannf., the dry root of radix codonpsis tangshen Codonopsis tangshen Oliv. and plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen.Its nature and flavor are sweet flat, nontoxic, and invigorating the spleen and replenishing QI is arranged, invigorating the spleen benefit lung effect.Cure mainly a little less than the spleen deficiency syndrome of the lung, the palpitaition of breathing hard, anorexia and loose stool, void is breathed with cough and is coughed, and interior heat disease such as quench one's thirst is the traditional tonic of China.But it is also common to pipe flower Radix Codonopsis Codonopsis tubelosa Kom. on market, the Codonopsis clematidea of codonopsis clematidea Clarke (Schyenk) C.B.Cl., gray wool Radix Codonopsis Codonopsiscanescens Nannf. and globe daisy Radix Codonopsis Codonopsis subglobosa W.W.Sm are sold as the puppet product as Radix Codonopsis surrogate and equal plant leopard Campanumoea javanica Blume ssp.japonica (Makino) Hong and balloonflower root Platycodon grandiflorum (Jacq.) A.DC..In Hong Kong, Radix Codonopsis is normally sold with trade(brand)name plate party, anti-party and line party.But the source, the specification difference, price difference is very big, with bad fill excellent common.In addition, the Radix Codonopsis that a kind of appearance scribbles red material is found on the Hongkong market first, and businessman is the stopping leak purpose with it.Be safe medication, we are by 5S rDNA gene sequencing technology and cooperate morphology, histology, HPLC-UV and X-Ray diffractive technology, and medicinal material itself and appearance material have been carried out deeply, studied in great detail.Found that this its plant origin of red Radix Codonopsis is plain flower Radix Codonopsis.Because geographical environment is planted method for planting, the variation of aspects such as upgrowth situation and complete processing has brought very big influence to the quality of quality, pharmaceutical intermediate and the Chinese patent medicine of codonopsis pilosula self.The traditional discrimination method of codonopsis pilosula is mainly based on form, micro-.Have no talent so far and find the compound that can be used as identifying mark in the chemistry aspect.Show that according to available data Radix Codonopsis at present more adopts control medicinal material to identify in contrast, does not obtain suitable reference substance so far.

How fast, accurately identify Radix Codonopsis source and all size, the true and false and quality, the existence of detection of a target Radix Codonopsis composition from the Radix Codonopsis product, it is one of important topic of the modernization of Chinese medicine that effective quality monitoring and quality evaluation carried out in Radix Codonopsis market.Though exploitation molecular engineering monitoring traditional Chinese medicine quality is at the early-stage, shown its broad prospect of application.In most of eukaryotes, rrna 5S rDNA gene is a series connection multiple multi-copy gene, and they independently exist, do not link to each other with main body RNA (18S-5.8S-28S rDNA), content is abundant in vivo, extensively distributes, and is called as biological intravital living fossil.In higher plant, each repeating unit of 5S rDNA gene is by transcriptional domain and transcribed spacer (Non transcribed spacer, NTS) form, the transcriptional domain high conservative, and transcribed spacer has bigger variation, the unification of this high conservative and variability, the useful mark that the molecule that makes 5S rDNA gene order become multi-copy gene in the research eukaryote forms and develops.The present invention utilizes the difference of 5S rDNA sequence to identify the Radix Codonopsis kind or distinguish its true and false just.For effectively controlling the labile factor in raw material sources and the production process, set about from genomics, with source, the true and false, the quality of identifying Radix Codonopsis kind medicinal material and the quality product of controlling Chinese patent medicine better, thereby, improve the safety and effectiveness of medicine for pharmaceutical industry provides scientific and reasonable guidance.

Summary of the invention:

A: the 5S rDNA sequence that codonopsis pilosula is provided.

B: utilize the kind of the 5S rDNA Sequence Identification stem of noble dendrobium and the method for telling truth from falsehood.

To achieve the object of the present invention, the contriver has collected 25 duplicate samples in different provinces from the whole nation, comprises 7 common Radix Codonopsis kinds in market, 3 commodity Radix Codonopsis, 3 specialty goods Radix Codonopsis and 2 pseudo-product kinds.All samples all passes through expert statement.The 5SrDNA sequence of 25 measured samples is as follows:

A. 7 kinds of Radix Codonopsis amount to 17 samples

(1) Radix Codonopsis Codonopsis pilosula (Franch.) Nannf. sample (numbering CMED-0038-04) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTGCTG GTATGCACCC

GTCAACTCCC CTCCTCTTTA TTCTTTGTAC TATCTCTAAA

CCATATAAGA AGGTCGTTTT GACTGCTCAC GCGCCCAAAA

CCGAGCAAAA CGACCTTTTC GCGCTCTTTC TTAATGTACG

CTGAGTAAAA AAAAGGAGGG AAGTTGACGG G

(2) Radix Codonopsis Codonopsis pilosula (Franch.) Nannf. sample (numbering CMED-0038-05) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTGCTG GTATGCACCC

GTCAAGTCCC CTCCTCTTTA TTGTTTGTAC TATCTCTAAA

CCATATAAGA AGGTCGTTTT GACTGCACAC GCGCCCAAAA

CCGAGCAAAA TGACCTTTTC GCGCTCTTTC TTAATGTACG

CTGAGTAAAA AAAAGGAGGG AAGTTGACGG G

(3) Radix Codonopsis Codonopsis pilosula (Franch.) Nannf. sample (numbering CMED-0038-08) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTGCTG GTATCCACCC

GTCAACTCCC CTCCTCTTTA TTCTTGGTAC TATCTCTAAA

CCATATAAGA AGGTCGTTAT GACTGCTGAC GCGCCCAAAA

GCGAGCAAAA CGTGCTTTTC GCGCTCTTAA TTATTGTACG

GTGAGCAAAA AAAAGGAGGG AAGTTGACGG G

(4) Radix Codonopsis Codonopsis pilsula (Franch.) Nannf. sample (numbering CMED-0038-12) 5S rDNA sequence:

CTAGGATGGG TGACCCCCAG GGAAGTGCTG GTATGCACCG

GTCAAGTCCC CTCCTCTTTA TTCTTTGTAA CATCTCTAAA

CCATATAAGA AGGTCGTTTT GACTGCTCAC GCGCCCAAAA

CCGAGCAAAA CGACCTTTTC GCGCTCTTGT TTAATGCGCG

CTGAGTAAAA AAAAGGAGGG AAGTTGACGG G

(5) plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-01) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCACC

CCCCTTTTGC TGTTTTTTTT ACTCAGCGTA CATTAAGAAA

CCATATAAAA GAAAGAAAAT GCTCGGTTTT GGGCGCGTGA

GCAGTCAAAA CGACCTTCTC GCGCTCTTTA GAGATAGTAC

AAAAGAATAA AGAGGAGGGA AGTTGACGGG

(6) plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-03) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCAGC

CCCCTTTTGC TCTTTTTTTT AATCAGCGTA CATTAACAAA

CCATATAAAA GAAAGAAAAT GCTCGGTTTT ACGCGCGTCA

GCAGTCAAAA CGACCTTCTC GCGCTCTTTA CACATAGTAC

AAAAGAATAA AGAGGTGGGA AGTTGACGGG

(7) plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-04) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCACC

CCCCTTTTGC TGTTTTTTT TACTCAGCCT ACTTTAAGAA

ACCATATAAA AGAAAGAAAA TCCTCGGTTT TGGGCGCGT

GAGCAGTCAA AACGACCTTC TCGCGCTCTT TAGAGATAGT

TCAAAAGAAT AAAGAGGAGG GAAGTTGACG GG

(8) plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-05) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCACC

CCCCTTTTGC TATTTTTTTT ACTCTGCCTA CATTAAGAAA

CGATATAAAA GAAAGAAAAT GCTCGGTTTT GGGCGAGTCA

GCAGTCAAAA GGACCTTCTC GCGGTCTTTA GAGATTGTAC

AAAAGAATAA AGCGGAGGGA AGTTGACGGG

(9) radix codonpsis tangshen Codonopsis tanghshen Oliv. sample (numbering CMED-0101-01) 5SrDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTCAACTCCC

CCCCCTTTTG CTGTTTTGTT TTACTCAGCG TACATTAAGA

AACCATATAA ACTGCTCACG CGCTCAAAAC CGGGCATTTT

CTTTCTCATA AACGACCTTC TAGTACGCTC TTTAGAGATA

GTACAAAAGA ATAAAGAGGA GGGAAGTTGA CGGG

(10) radix codonpsis tangshen Codonopsis tangshen Oliv. sample (numbering CMED-0101-02) 5SrDNA sequence:

CTAGGATGGG TGACCCCCTG GGAACCCCTC GTCAAGTCCC

CCCCCTTTTG GTGTTTTGTT TTACTCACCG TACATTAAGA

AACCTTATAA AGTGCTCACG CGGTCAAAAC CGGCCATTTT

CTTTCTCAAA AACGACCTTC TAGTACGGGC TTTAGAGATA

CTACAAAAGA ATAAAGAGCA GGGAAGTTGA CGGG

(11) radix codonpsis tangshen Codonopsis tangshen Oliv. sample (numbering CMED-0101-05) 5SrDNA sequence:

CTAGGATGGG TGGCCCCCTG GCAAGTCCTC GTCAAGTCCC

CCCCCTTTTG GTGTTTTGTT TTTCTCAGCG TACGTTAAGA

AACCATATAA AGGGCTCACG CGGTCAAAAC CGGGCGTTTT

CTTTCTAATA AACTGCCTTC TAGTAGGCTC TTTAGACATA

GTAGAAAAGA ATAAAGAGCA GGGAAGTTGA CGGG

(12) pipe flower Radix Codonopsis Codonopsis tubelosa Kom. sample (numbering CMED-0102-01) 5S rDNA sequence:

CTAGGACTGC TCACGGCCCG AAAAGTCCGA GCATTTTTCT

TTCTTTTATA CTGGTTTCTT AATGTACGGT GAGTAAAAAA

ACAGCAAAAG GGGGGGTGCA ACACGAGGTT CCCAGGGGGT

CACCCATCCT AATACTACTC TCGCCCTCAT TTGAGTTCGT

ACAAAACGAC CATATGGGAG AGGAGTTGAC GGG

(13) pipe flower Radix Codonopsis Codonopsis tubelosa Kom. sample (numbering CMED-0102-02) 5S rDNA sequence:

CTAGGACTGC TCACGGCCCG AAAAGTCCTA GCATGTTTCT

TTCTTTTATA CTGTTTTCTT ACTGTACGGT GACTAAAAAA

ACAGCTAAAG GGGGGGTGCT ACACGAGGTC GCCAGGGGGA

CACGCATCCT AATACGACTC TCTCCCTCAT TTTAGTTCGT

ACAAAACTAC AAAAAGGGAG AGAAGTTGAC GGG

(14) Codonopsis of codonopsis clematidea Clarke clematidea (Schyenk) C.B.Cl. sample (numbering CMED-0109-01) 5S rDNA sequence:

CTAGGATGGT CGACCATATG GGAAAGCTTC GCGTTTTTCT

CCCTTTTTGA GTTTTTTTTA GTGTCACCTA AATAAGGAAA

CCATTAAAAG AAAGAAACTA CACGAGGTTT TGGGGCCGTT

ATCCGCTCAC AAACTATTCT GGCATTCGTT TGGGATAGCA

TAAAAGACTA AAGCCGGGGG GAGTTGACGG G

(15) gray wool Radix Codonopsis Codonopsis canescens Nannf. sample (numbering CMED-0110-01) 5S rDNA sequence:

CTAGGACGCG GGAATTCGAG AAGAGTGCTG GTATGATCGC

ACCCGCTGTT TTTTTTCCTC TTTATTCTTT TGTACTATCT

CTAAAGAGCG CGAGAAGGTC CGGTTTTGAC TTCTCACGCG

CCCAAAACAA AACGAGCTTT TTGCCTTTCG TTTGAGACGG

TATAAAAGTA CGCTGAGGAG GGGAGTTGAC GGG

(16) globe daisy Radix Codonopsis Codonopsis subglobosa W.W.Sm sample (numbering CMED-0107-01) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTATGCACCC

GTTAACCCCC CCTTTTTTTG TTTTTTTTAC TATCTCTACG

TTAAGAAAAT GCTCGGTTTG ACTGCTCATG GGCGCGTGAG

CAGTCAAAAC GACCTTCTCG CGCTCTTTAG AGATATGTAC

AAAAGAATAA AGAGGAGGGG AGTTGACGGG

(17) globe daisy Radix Codonopsis Codonopsis subglobosa W.W.Sm sample (numbering CMED-0107-02) 5S rDNA sequence:

CTAGGATGGG TGTCCCCCTG GGAAGACCTC GTATGCACCC

CCTAACCCCC CCTTTTTTTG TTTTTTTTAC TATCGCAACG

TTAAGAAAAT GCTTCCTTTG ACTGCTGATG GGCGCGTCAG

CAATCAAAAC GTCCTTCTCG CGCACTTTAG AGATATGGAC

AAAAGAATAA AGAGGACGGG TGTTGACGGG

B. 3 samples of commodity Radix Codonopsis

(18) the Radix Codonopsis of plate party sample (numbering CMED-DS-8-DFH) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTTTTA GTCAACTATG

CCCGCTTTTG CTGTTTTGTT TTCCTCAGCG TTCTTTAAGA

ATCCATAAAG ACTGCTCACG GGCTCAAAAC CGGGCATTTT

CTTTCCCAAA ACCGAGCTTC TAGTTCGCTC TTTAGAGATA

GTACAAAAGA ATAAAGAGGA GGGAAGTTGA CGGG

(19) the anti-Radix Codonopsis of party sample (numbering CMED-DS-9-YH) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAATCTTC GTGTTGCACC

CCCCTTTTGC TGTTTTTTTT ACTAAACGGA CATTAAGAAG

CGAATAAAAG AAAGAAAATG CTCGGTTTTG AGCGCGAGAG

CAGTCAAAAA GAACTTCTCG CGGTTGTTTG AGATAGTATA

AAAGAATAAA GAGGAGGGAA GTTGACGGG

(20) the Radix Codonopsis of line party sample (numbering CMED-DS-10-YRS) 5SrDNA sequence:

CTAGGATGGG TGACTCCCTG GGAAGTCCTC GTGTTGCACC

CCCCTTTTGC TGTTTTTTTT ACTCACCGTA CATTAAGAAA

CCATATAAAA GAAAGAAAAA TGCTCGGTTT TGGGGCCGTG

AGCAGTCAAA ACGACCTTCT GGCGCTCGTT TGAGATAGTA

CAAAAGAATA AAGAGGAGGG GAGTTGACGG G

C. 3 samples are joined in the specialty goods Colorado Party

(21) Colorado Party ginseng Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-DS-7-XH) 5S rDNA sequence:

CTAGGATGGA TGACCCCCTG GGAAGTCCTC GTGTTGCACC

CCCCCTTTTG GTGTTTTTTT TACTCTGCGT ACATTAAGAA

ACGATATAAA AGAAAGAAAA TGCTGGGTTT TGGGCGCGTG

AGCAGTCAGA ACGACCTTGT CGCGCTCTTT AGAGATAGTA

CAAAAGAATA AAGAGGAGGG GAGTTGACGG G

(22) Colorado Party ginseng Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-DS-11-WYT) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCGTC GTGTTGCACC

CCCCCTTTTG CTGTTTTTTT TACTGAGCGT ACATTAAGAA

ACCATGTAAA AGAAAGAAAA TGCTCGGTTT TGGGCCCGTG

AGCAGTCAAA ACGACGTTCT CGCGCTCTTT GAGATAGTAC

AAAAGAATAA AGAGGAGGGG AGTTGACGGG

(23) Colorado Party ginseng Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-DS-12-YRS) 5S rDNA sequence:

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTACACC

CCCCTTTTGC TGTTTTTTTT GCTCAGCGTA CATTAAGAAA

TCATGTAAAA GAAAGAAAAT GCTCGGTTTT GGGCGCCTGA

GCAGTCAAAA CGACCTTCTC GCGGTCTTTA GAGATAGTAC

AAAAGAATAA AGAGGAGGGG AGTTGACGGG

D. the pseudo-product leopard of Radix Codonopsis and 2 kinds of balloonflower root

(24) leopard Campanumoea javanica Blume ssp.japonica (Makino) Hong sample (numbering CMED-0108-01) 5S rDNA sequence:

CTAGGATGGG AGACACCCTG GTAAATCTTC GTGTTGCAAT

TCCCCTTTTG CTGTTTTTTT TACTAAACGG AGGTCTAGAA

GCGAATAAAA GAAAGAAAGA ATAACCATTT TTGAGGCTGA

GGTAAGGTCT GAAGAACTTC TGAAGGTTGT TTGAGGCCGT

ATAAAAATGT TCGAACAAGG GGAGTTGACG GG

(25) balloonflower root Platycodon grandiflorum (Jacq.) A.DC. sample (numbering CMED-0111-01) 5S rDNA sequence:

CTAGGATGGG TGACCTCCTG GGAAGGCCTC CTGTTGCACC

CCCCATTTTT TTTATTTAAT TTTTGATTTA TTTATGTTAT

CGGTAGGTCG TTTTTTTTGT AGCTGTTGGA ATCATATTGC

GTTTTGTGAG TGCGTTTATC CGTAATAATA GGGAGCGTAA

CGGTAAGGTT TCGGGTGTCG GGTCCGTAAC GGTTGAAACG

GGGAATCGTA ATGAGGGAAA ACGGTGGAAT TTCGGTTAAA

ATGGTAAAAG GATGTTGAAT AAAATGCAAT TTATGGTGAG

AGTATATGGG

Embodiment:

The measuring method of the 5S rDNA sequence of above-mentioned 25 samples comprises the following steps:

(1) collects plant sample, carry out cultivar identification work;

(2) from the plant that step (1) is selected, extract DNA;

(3) DNA that extracts with step (2) is a template, with the primer of oligonucleotide as polymerase chain reaction (PCR), and amplification 5S rDNA section, used primer picks up from the conservative section at plant 5SrDNA two ends respectively;

(4) using the cloning and sequencing technology checks order to the PCR product that is obtained in the step (3).

Each sample repeat at least to check order 3 times and the sequence of sample also internet page National Center for Biotechnology Information (NCBI) and EuropeanMolecular Biology Laboratory (EMBL) and other plant sequence make comparisons and verify to guarantee 25 sequences of the present invention accurate.

Mean difference at dna level between the sample of above-mentioned measured same kind is 6.2% to 9.9%, and the difference of 5S rDNA sequence is 26% to 60% between described Radix Codonopsis kind.And the difference of the 5S rDNA sequence of Radix Codonopsis and Campanumoea javanica is 37% to 57%.The difference of the 5S rDNA sequence of Radix Codonopsis and balloonflower root is 63% to 72%.

The present invention utilizes in the difference of 5S rDNA sequence plant in the Radix Codonopsis class plant and the comparison between kind, identifies different Radix Codonopsis kinds whereby and distinguishes its true and false.The method that is adopted comprises the following steps:

1. from plant sample, extract DNA;

2. the DNA that extracts with step 1 is a template, with oligonucleotide as polymerase chain reaction (PCR) primer, amplification 5S rDNA section, used primer picks up from the conservative section at plant 5S rDNA two ends respectively;

3. using the cloning and sequencing technology checks order to the PCR product that step 2 obtained;

4. the 5S rDNA sequence comparison of the dna sequence dna that step 3 is recorded and above-mentioned 18 Radix Codonopsis and Campanumoea javanica is with the Radix Codonopsis kind of identifying institute's measuring plants and tell truth from falsehood.

Describe described method in detail:

(1) extraction of DNA

DNA extraction method of plant and Draper and Scott (Draper and Scott, 1988) are described roughly the same, are summarized as follows: the exsiccant vegetable material is washed with 70% alcoholic acid distilled water solution, then, mix and grind into powder with liquid nitrogen.The powdered sample of gained and the CTAB of 6 times of volumes are extracted the damping fluid mixing, and insulation added chloroform-primary isoamyl alcohol (24: 1) mixed solution extracting of equivalent after 30 minutes in 56 ℃ of water-baths, and is centrifugal, gets supernatant liquor.The 10%CTAB solution that adds 0.1 volume in resistates, room temperature is placed after 10 minutes and extracted once with chloroform-primary isoamyl alcohol, and is centrifugal, gets supernatant liquor and mixes with last time supernatant liquor.The CTAB precipitation buffering liquid that adds equivalent in the gained supernatant liquor produces precipitation, and room temperature was placed after 1 hour with 13000g centrifugal 15 minutes.Be dissolved in the TE damping fluid (10mMTris-HCl, pH 7.5,1mM EDTA) after with ethanol purification the DNA throw out standby.

(2) pcr amplification reaction

The DNA of 5S rDNA section is obtained by pcr amplification reaction.The primer that reacts used is 5S2F and 5S2R:

The 5S2F sequence is: 5 ' GTG CTT GGG CGA GAG TAG TA

The 5S2R sequence is: 5 ' TTA GTG CTG GTA TGA TCG CA

Reaction soln comprises: 10ng DNA of plants, 1XTag buffered soln (10mMTris-HCl, pH8.3,0.001% gelatin), 0.2mM dNTPs, 1 μ M primer, and the Taq DNA polymerase of 1 unit.Reaction conditions is: in 94 ℃ of pre-treatment 5 minutes, carry out 35 circulations then, cycling condition is: 94 ℃, and 1 minute, 60 ℃, 1 minute, 72 ℃, 2 minutes; Last 72 ℃, 10 minutes.

(3) cloning and sequencing technology

(Bio 101, USA) carry out purifying with DNA purification kit GENCLEAN IIIKit for the PCR product that step (2) is obtained.Take out the PCR fragment of 1 μ l (about 50ng) purifying, with clone test kit TA cloning Kit (pGEM-T Easy Vector Systems, Promega, USA) with this PCR fragment cloning to carrier.After transformant is cultivated, with plasmid purification test kit Rapid Plasmid Miniprep System (Marligen Bioscience, Germany) plasmid purification.Order-checking is by reaction kit ABI Prism BigDye Terminator CycleSequencing Ready Reaction Kit (PE Biosystems, USA) and genetic analyzer Genetic analyzer (ABI Prism 3100, Applied Biosystems USA) carries out.(Perkin Elmer is USA) with DNAsis version 7.00 (Hitachi, Japan) dna sequence dna of analysis and definite sample to use sequence analysis software Sequencing Analysis version 3.0 at last.

(4) dna sequence dna relatively

Sequence is stored with the Fasta form, use sequence analysis software BioEdit version7.0.1 (Isis Pharmaceuticals then, USA) sort and calculated difference per-cent, the result imports Excel 2002 (Microsoft Corporation, USA) analysis that takes statistics again.

(5) evaluation of tested plant sample

The 5S rDNA sequence of institute's test sample product and the DNA standard of all Radix Codonopsis kinds disclosed by the invention are compared as stated above one by one.As find that the sequence of this plant sample and certain Radix Codonopsis 5S rDNA sequence calibration are complementary, that is to say that its sequence difference is less than 9.9%, i.e. tested plant sample of susceptible of proof and Radix Codonopsis, and with calibration (Radix Codonopsis) be same kind.On the contrary, as all not being complementary by the 5S rDNA sequence of measuring plants and any one Radix Codonopsis standard DNA sequence, then this tested plant variety of explanation is not a Radix Codonopsis.

If which kind of pseudo-product or surrogate thinks further to understand this is by the measuring plants product, then the sequence of its sequence and several pseudo-product disclosed by the invention and surrogate can be compared, as finding coupling, illustrate promptly this sample is to belong to which kind of pseudo-product or surrogate.

Therefore, adopt dna sequence dna and the authentication method of the 5S rDNA of Radix Codonopsis provided by the invention, the comparison of the dna sequence dna by its 5S rDNA makes convenient and reliable to the evaluation of Chinese medicine Radix Codonopsis.

Sequence table

<210>SEQ?ID?NO：1

<211>LENGTH：210

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?(Franch.)?Nannf?1

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：1

CTAGGATGGG TGACCCCCTG GGAAGTGCTG GTATGCACCC GTCAACTCCC CTCCTCTTTA

60

TTCTTTGTAC TATCTCTAAA CCATATAAGA AGGTCGTTTT GACTGCTCAC GCGCCCAAAA

120

CCGAGCAAAA CGACCTTTTC GCGCTCTTTC TTAATGTACG CTGAGTAAAA AAAAGGAGGG

180

AAGTTGACGG G

191

<210>SEQ?ID?NO：2

<211>LENGTH：211

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?(Franch.)?Nannf?2

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：2

CTAGGATGGG TGACCCCCTG GGAAGTGCTG GTATGCACCC GTCAAGTCCC CTCCTCTTTA

60

TTGTTTGTAC TATCTCTAAA?CCATATAAGA AGGTCGTTTT GACTGCACAC GCGCCCAAAA

120

CCGAGCAAAA?TGACCTTTTC GCGCTCTTTC TTAATGTACG CTGAGTAAAA AAAAGGAGGG

180

AAGTTGACGG G

191

<210>SEQ?ID?NO：3

<211>LENGTH：210

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?(Franch.)?Nannf?3

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：3

CTAGGATGGG TGACCCCCTG GGAAGTGCTG GTATCCACCC GTCAACTCCC CTCCTCTTTA

60

TTCTTGGTAC TATCTCTAAA CCATATAAGA AGGTCGTTAT GACTGCTGAC GCGCCCAAAA

120

GCGAGCAAAA CGTGCTTTTC GCGCTCTTAA TTATTGTACG GTGAGCAAAA AAAAGGAGGG

180

AAGTTGACGG G

191

<210>SEQ?ID?NO：4

<211>LENGTH：210

<212>TYpE：DNA

<213>ORGANISM：Codonopsis?pilosula?(Franch.)?Nannf?4

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：4

CTAGGATGGG TGACCCCCAG GGAAGTGCTG GTATGCACCG GTCAAGTCCC CTCCTCTTTA

60

TTCTTTGTAA CATCTCTAAA CCATATAAGA AGGTCGTTTT GACTGCTCAC GCGCCCAAAA

120

CCGAGCAAAA CGACCTTTTC GCGCTCTTGT TTAATGCGCG CTGAGTAAAA AAAAGGAGGG

180

AAGTTGACGG G

191

<210>SEQID?NO：5

<211>LENGTH：208

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?var.?Modesta?(Nannf.)?L.T.Shen?5

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：5

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCACC CCCCTTTTGC TGTTTTTTTT

60

ACTCAGCGTA CATTAAGAAA CCATATAAAA GAAAGAAAAT GCTCGGTTTT GGGCGCGTGA

120

GCAGTCAAAA CGACCTTCTC GCGCTCTTTA GAGATAGTAC AAAAGAATAA AGAGGAGGGA

180

AGTTGACGG G

190

<210>SEQ?ID?NO：6

<211>LENGTH：208

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?var.?Modesta?(Nannf.)?L.T.Shen?6

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：6

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCAGC CCCCTTTTGC TCTTTTTTTT

60

AATCAGCGTA CATTAACAAA CCATATAAA GAAAGAAAT GCTCGGTTTT ACGCGCGTCA

120

GCAGTCAAAA CGACCTTCTC GCGCTCTTTA CACATAGTAC AAAAGAATAA AGAGGTGGGA

180

AGTTGACGGG

190

<210>SEQID?NO：7

<211>LENGTH：209

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?var.?Modesta?(Nannf.)?L.T.Shen?7

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：7

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCACC CCCCTTTTGC TGTTTTTTTT

60

ACTCAGCCTA CTTTAAGAAA CCATATAAAA GAAAGAAAAT CCTCGGTTTT GGGCGCGTGA

120

GCAGTCAAAA CGACCTTCTC GCGCTCTTTA GAGATAGTTC AAAAGAATAA AGAGGAGGGA

180

AGTTGACGGG

190

<210>SEQ?ID?NO：8

<211>LENGTH：208

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?var.?Modesta?(Nannf.)?L.T.Shen?8

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：8

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTGCACC CCCCTTTTGC TATTTTTTTT

60

ACTCTGCCTA CATTAAGAAA CGATATAAAA GAAAGAAAAT GCTCGGTTTT GGGCGAGTCA

120

GCAGTCAAAA GGACCTTCTC GCGGTCTTTA GAGATTGTAC AAAAGAATAA AGCGGAGGGA

180

AGTTGACGGG

190

<210>SEQ?ID?NO：9

<211>LENGTH：213

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?tangshen?Oliv.9

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：9

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTCAACTCCC CCCCCTTTTG CTGTTTTGTT

60

TTACTCAGCG TACATTAAGA AACCATATAA ACTGCTCACG CGCTCAAAAC CGGGCATTTT

120

CTTTCTCATA AACGACCTTC TAGTACGCTC TTTAGAGATA GTACAAAAGA ATAAAGAGGA

180

GGGAAGTTGA CGGG

194

<210>SEQID?NO：10

<211>LENGTH：213

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?tangshen?Oliv.10

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：10

CTAGGATGGG TGACCCCCTG GGAACCCCTC GTCAAGTCCC CCCCCTTTTG GTGTTTTGTT

60

TTACTCACCG TACATTAAGA AACCTTATAA AGTGCTCACG CGGTCAAAAC CGGCCATTTT

120

CTTTCTCAAA AACGACCTTC TAGTACGGGC TTTAGAGATA CTACAAAAGA ATAAAGAGCA

180

GGGAAGTTGA CGGG

194

<210>SEQID?NO：11

<211>LENGTH：213

<212>TYPE：DNA

<213>ORGANISM：Codonopsis tangshen?Oiv.11

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：11

CTAGGATGGG TGGCCCCCTG GCAAGTCCTC GTCAAGTCCC CCCCCTTTTG GTGTTTTGTT

60

TTTCTCAGCG TACGTTAAGA AACCATATAA AGGGCTCACG CGGTCAAAAC CGGGCGTTTT

120

CTTTCTAATA AACTGCCTTC TAGTAGGCTC TTTAGACATA GTAGAAAAGA ATAAAGAGCA

180

GGGAAGTTGA CGGG

194

<210>SEQID?NO：12

<211>LENGTH：212

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?tubelosa?Kom.12

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：12

CTAGGACTGC TCACGGCCCG AAAAGTCCGA GCATTTTTCT TTCTTTTATA CTGGTTTCTT

60

AATGTACGGT GAGTAAAAAA ACAGCAAAAG GGGGGGTGCA ACACGAGGTT CCCAGGGGGT

120

CACCCATCCT AATACTACTC TCGCCCTCAT TTGAGTTCGT ACAAAACGAC CATATGGGAG

180

AGGAGTTGAC GGG

193

<210>SEQ?ID?NO：13

<211>LENGTH：212

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?tubelosa?Kom.13

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：13

CTAGGACTGC TCACGGCCCG AAAAGTCCTA GCATGTTTCT TTCTTTTATA CTGTTTTCTT

60

ACTGTACGGT GACTAAAAAA ACAGCTAAAG GGGGGGTGCT ACACGAGGTC GCCAGGGGGA

120

CACGCATCCT AATACGACTC TCTCCCTCAT TTTAGTTCGT ACAAAACTAC AAAAAGGGAG

180

AGAAGTTGAC GGG

193

<210>SEQID?NO：14

<211>LENGTH：210

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?clematidea?(Schyenk)?C.B.Cl.14

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：14

CTAGGATGGT CGACCATATG GGAAAGCTTC GCGTTTTTCT CCCTTTTTGA GTTTTTTTTA

60

GTGTCACCTA AATAAGGAAA CCATTAAAAG AAAGAAACTA CACGAGGTTT TGGGGCCGTT

120

ATCCGCTCAC AAACTATTCT GGCATTCGTT TGGGATAGCA TAAAAGACTA AAGCCGGGGG

180

GAGTTGACGG G

191

<210>SEQID?NO：15

<211>LENGTH：212

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?canescens?Nannf.15

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：15

CTAGGACGCG GGAATTCGAG AAGAGTGCTG GTATGATCGC ACCCGCTGTT TTTTTTCCTC

60

TTTATTCTTT TGTACTATCT CTAAAGAGCG CGAGAAGGTC CGGTTTTGAC TTCTCACGCG

120

CCCAAAACAA AACGAGCTTT TTGCCTTTCG TTTGAGACGG TATAAAAGTA CGCTGAGGAG

180

GGGAGTTGAC GGG

193

<210>SEQID?NO：16

<211>LENGTH：208

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?subglobosa?W.W.Sm?16

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：16

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTATGCACCC GTTAACCCCC CCTTTTTTTG

60

TTTTTTTTAC TATCTCTACG TTAAGAAAAT GCTCGGTTTG ACTGCTCATG GGCGCGTGAG

120

CAGTCAAAAC GACCTTCTCG CGCTCTTTAG AGATATGTAC AAAAGAATAA AGAGGAGGGG

180

AGTTGACGGG

190

<210>SEQID?NO：17

<211>LENGTH：208

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?subglobosa?W.W.Sm?17

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：17

CTAGGATGGG TGTCCCCCTG GGAAGACCTC GTATGCACCC CCTAACCCCC CCTTTTTTTG

60

TTTTTTTTAC TATCGCAACG TTAAGAAAAT GCTTCCTTTG ACTGCTGATG GGCGCGTCAG

120

CAATCAAAAC GTCCTTCTCG CGCACTTTAG AGATATGGAC AAAAGAATAA AGAGGACGGG

180

TGTTGACGGG

190

<210>SEQID?NO：18

<211>LENGTH：213

<212>TYPE：DNA

<213>ORGANISM：Radix?Codonopsis?18

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：18

CTAGGATGGG TGACCCCCTG GGAAGTTTTA GTCAACTATG CCCGCTTTTG CTGTTTTGTT

60

TTCCTCAGCG TTCTTTAAGA ATCCATAAAG ACTGCTCACG GGCTCAAAAC CGGGCATTTT

120

CTTTCCCAAA ACCGAGCTTC TAGTTCGCTC TTTAGAGATA GTACAAAAGA ATAAAGAGGA

180

GGGAAGTTGA CGGG

194

<210>SEQ?ID?NO：19

<211>LENGTH：207

<212>TYPE：DNA

<213>ORGANISM：Radix?Codonopsis?19

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：19

CTAGGATGGG TGACCCCCTG GGAAATCTTC GTGTTGCACC CCCCTTTTGC TGTTTTTTTT

60

ACTAAACGGA CATTAAGAAG CGAATAAAAG AAAGAAAATG CTCGGTTTTG AGCGCGAGAG

120

CAGTCAAAAA GAACTTCTCG CGGTTGTTTG AGATAGTATA AAAGAATAAA GAGGAGGGAA

180

GTTGACGGG

189

<210>SEQID?NO：20

<211>LENGTH：210

<212>TYPE：DNA

<213>ORGANISM：Radix?Codonopsis?20

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：20

CTAGGATGGG TGACTCCCTG GGAAGTCCTC GTGTTGCACC CCCCTTTTGC TGTTTTTTTT

60

ACTCACCGTA CATTAAGAAA CCATATAAAA GAAAGAAAAA TGCTCGGTTT TGGGGCCGTG

120

AGCAGTCAAA ACGACCTTCT GGCGCTCGTT TGAGATAGTA CAAAAGAATA AAGAGGAGGG

180

GAGTTGACGG G

191

<210>SEQ?ID?NO：21

<211>LENGTH：210

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?var.?Modesta?(Nannf.)?L.T.Shen?21

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：21

CTAGGATGGA TGACCCCCTG GGAAGTCCTC GTGTTGCACC CCCCCTTTTG GTGTTTTTTT

60

TACTCTGCGT ACATTAAGAA ACGATATAAA AGAAAGAAAA TGCTGGGTTT TGGGCGCGTG

120

AGCAGTCAGA ACGACCTTGT CGCGCTCTTT AGAGATAGTA CAAAAGAATA AAGAGGAGGG

180

GAGTTGACGG G

191

<210>SEQ?ID?NO：22

<211>LENGTH：208

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?var.?Modesta?(Nannf.)?L.T.Shen?22

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：22

CTAGGATGGG TGACCCCCTG GGAAGTCGTC GTGTTGCACC CCCCCTTTTG CTGTTTTTTT

60

TACTGAGCGT ACATTAAGAA ACCATGTAAA AGAAAGAAAA TGCTCGGTTT TGGGCCCGTG

120

AGCAGTCAAA ACGACGTTCT CGCGCTCTTT GAGATAGTAC AAAAGAATAA AGAGGAGGGG

180

AGTTGACGGG

190

<210>SEQ?ID?NO：23

<211>LENGTH：208

<212>TYPE：DNA

<213>ORGANISM：Codonopsis?pilosula?var.?Modesta?(Nannf.)?L.T.Shen?23

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：23

CTAGGATGGG TGACCCCCTG GGAAGTCCTC GTGTTACACC CCCCTTTTGC TGTTTTTTTT

60

GCTCAGCGTA CATTAAGAAA TCATGTAAAA GAAAGAAAAT GCTCGGTTTT GGGCGCCTGA

120

GCAGTCAAAA CGACCTTCTC GCGGTCTTTA GAGATAGTAC AAAAGAATAA AGAGGAGGGG

180

AGTTGACGGG

190

<210>SEQ?ID?NO：24

<211>LENGTH：211

<212>TYPE：DNA

<213>ORGANISM：Campanumoea?javanica?Blume?ssp.?japonica?(Makino)?Hong?24

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：24

CTAGGATGGG AGACACCCTG GTAAATCTTC GTGTTGCAAT TCCCCTTTTG CTGTTTTTTT

60

TACTAAACGG AGGTCTAGAA GCGAATAAAA GAAAGAAAGA ATAACCATTT TTGAGGCTGA

120

GGTAAGGTCT GAAGAACTTC TGAAGGTTGT TTGAGGCCGT ATAAAAATGT TCGAACAAGG

180

GGAGTTGACG GG

192

<210>SEQ?ID?NO：25

<211>LENGTH：319

<212>TYPE：DNA

<213>ORGANISM：Platycodon?grandiflorum?(Jacq.)?A.DC.25

<223>OTHER?INFORMATION：5S?rDNA?Spacer

<400>SEQUENCE：25

CTAGGATGGG TGACCTCCTG GGAAGGCCTC CTGTTGCACC CCCCATTTTT TTTATTTAAT

60

TTTTGATTTA TTTATGTTAT CGGTAGGTCG TTTTTTTTGT AGCTGTTGGA ATCATATTGC

120

GTTTTGTGAG TGCGTTTATC CGTAATAATA GGGAGCGTAA CGGTAAGGTT TCGGGTGTCG

180

GGTCCGTAAC GGTTGAAACG GGGAATCGTA ATGAGGGAAA ACGGTGGAAT TTCGGTTAAA

240

ATGGTAAAAG GATGTTGAAT AAAATGCAAT TTATGGTGAG AGTATATGGG

290

Claims (9)

1. method of identifying the Radix Codonopsis kind, it is characterized in that, use one or more dna sequence dnas that are selected from the calibration group, the 5S rDNA of sample to be identified is compared as calibration, thereby obtain the sequence difference per-cent of this sample and calibration, used calibration group comprises following dna sequence dna:

(1) the 5S rDNA sequence of Radix Codonopsis Codonopsis pilosula (Franch.) Nannf. sample (numbering CMED-0038-04), this sequence is shown in SEQ ID NO.1;

(2) the 5S rDNA sequence of Radix Codonopsis Codonopsis pilosula (Franch.) Nannf. sample (numbering CMED-0038-05), this sequence is shown in SEQ ID NO.2;

(3) the 5S rDNA sequence of Radix Codonopsis Codonopsis pilosula (Franch.) Nannf. sample (numbering CMED-0038-08), this sequence is shown in SEQ ID NO.3;

(4) the 5S rDNA sequence of Radix Codonopsis Codonopsis pilosula (Franch.) Nannf. sample (numbering CMED-0038-12), this sequence is shown in SEQ ID NO.4;

(5) the 5S rDNA sequence of plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-01), this sequence is shown in SEQ IDNO.5;

(6) the 5S rDNA sequence of plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-03), this sequence is shown in SEQ IDNO.6;

(7) the 5S rDNA sequence of plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-04), this sequence is shown in SEQ IDNO.7;

(8) the 5S rDNA sequence of plain flower Radix Codonopsis Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-0093-05), this sequence is shown in SEQ IDNO.8;

(9) the 5S rDNA sequence of radix codonpsis tangshen Codonopsis tangshen Oliv. sample (numbering CMED-0101-01), this sequence is shown in SEQ ID NO.9;

(10) the 5S rDNA sequence of radix codonpsis tangshen Codonopsis tangshen Oliv. sample (numbering CMED-0101-02), this sequence is shown in SEQ ID NO.10;

(11) the 5S rDNA sequence of radix codonpsis tangshen Codonopsis tangshen Oliv. sample (numbering CMED-0101-05), this sequence is shown in SEQ ID NO.11;

(12) the 5S rDNA sequence of pipe flower Radix Codonopsis Codonopsis tubelosa Kom. sample (numbering CMED-0102-01), this sequence is shown in SEQ ID NO.12;

(13) the 5S rDNA sequence of pipe flower Radix Codonopsis Codonopsis tubelosa Kom. sample (numbering CMED-0102-02), this sequence is shown in SEQ ID NO.13;

(14) the 5S rDNA sequence of the Codonopsis of codonopsis clematidea Clarke clematidea (Schyenk) C.B.C1. sample (numbering CMED-0109-01), this sequence is shown in SEQ ID NO.14;

(15) the 5S rDNA sequence of gray wool Radix Codonopsis Codonopsis canescens Nannf. sample (numbering CMED-0110-01), this sequence is shown in SEQ ID NO.15;

(16) the 5S rDNA sequence of globe daisy Radix Codonopsis Codonopsis subglobosa W.W.Sm sample (numbering CMED-0107-01), this sequence is shown in SEQ ID NO.16;

(17) the 5S rDNA sequence of globe daisy Radix Codonopsis Codonopsis subglobosa W.W.Sm sample (numbering CMED-0107-02), this sequence is shown in SEQ ID NO.17;

(18) the 5S rDNA sequence of the Radix Codonopsis of plate party sample (numbering CMED-DS-8-DFH), this sequence is shown in SEQ ID NO.18;

(19) the 5SrDNA sequence of the anti-Radix Codonopsis of party sample (numbering CMED-DS-9-YH), this sequence is shown in SEQ ID NO.19;

(20) the 5S rDNA sequence of the Radix Codonopsis of line party sample (numbering CMED-DS-10-YRS), this sequence is shown in SEQ ID NO.20;

(21) the 5S rDNA sequence of Colorado Party ginseng Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-DS-7-XH), this sequence is shown in SEQ ID NO.21;

(22) the 5S rDNA sequence of Colorado Party ginseng Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-DS-11-WYT), this sequence is shown in SEQ IDNO.22;

(23) the 5S rDNA sequence of Colorado Party ginseng Codonopsis pilosula var.Modesta (Nannf.) L.T.Shen sample (numbering CMED-DS-12-YRS), this sequence is shown in SEQ IDNO.23;

(24) the 5S rDNA sequence of leopard Campanumoea javanica Blume ssp.japonica (Makino) Hong sample (numbering CMED-0108-01), this sequence is shown in SEQ IDNO.24; With

(25) the 5S rDNA sequence of balloonflower root Platycodon grandiflorum (Jacq.) A.DC. sample (numbering CMED-0111-01), this sequence is shown in SEQ ID NO.25.

2. the method for claim 1, it is characterized in that, further comprise a determining step, this determining step is according to a difference thresholding (threshold), sequence difference per-cent can tentatively judge then that less than this thresholding this sample and calibration are same kinds between sample and calibration.

3. method as claimed in claim 2 is characterized in that, described difference thresholding is between 8% to 12%.

4. method as claimed in claim 3 is characterized in that, described difference thresholding is 9.9%.

5. the method for claim 1 is characterized in that, this method comprises following concrete steps:

(a) from sample to be identified, extract DNA;

(b) be template with above-mentioned DNA, and be primer, use polymerase chain reaction (PCR) amplification 5S rDNA section with suitable oligonucleotide;

(c) the 5S rDNA section to above-mentioned amplification carries out sequencing;

(d) above-mentioned institute test sample product sequence and one or more dna sequence dnas that is selected from the calibration group are compared, and calculate difference percentage between it; With

(e) according to above-mentioned difference percentage judge this identified sample whether be same sample than calibration.

6. method as claimed in claim 5 is characterized in that, described primer is 5S2F, and its sequence is 5 ' GTG CTT GGG CGA GAG TAG TA and 5S2R, and its sequence is 5 ' TTA GTG CTG GTA TGA TCG CA.

7. method as claimed in claim 6 is characterized in that, the judgement of step (e) realizes that according to a difference thresholding sequence difference per-cent can tentatively judge then that less than this thresholding this sample and calibration are same kinds between sample and calibration.

8. method as claimed in claim 7 is characterized in that, described difference thresholding is between 8% to 12%.

9. method as claimed in claim 8 is characterized in that, described difference thresholding is 9.9%.