CN107400723A - The authentication method and purposes of seed plant species - Google Patents
The authentication method and purposes of seed plant species Download PDFInfo
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- CN107400723A CN107400723A CN201710833314.3A CN201710833314A CN107400723A CN 107400723 A CN107400723 A CN 107400723A CN 201710833314 A CN201710833314 A CN 201710833314A CN 107400723 A CN107400723 A CN 107400723A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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Abstract
The present invention relates to a kind of authentication method and purposes of seed plant species, comprise the following steps：Selected seed plant species, obtain the ITS2 gene orders of the seed plant species, chadogram is built based on ITS2 gene orders, multipair sibling species are chosen in chadogram, calculate the genetic distance between each pair sibling species, significance test of difference is carried out to the multigroup genetic distance for belonging to same door species, the inapparent genetic distance calculating of difference is chosen and is averagely worth to molecule dividing value；The genetic distance between species and target species to be measured is calculated, the genetic distance is determined into species classification compared with the molecule dividing value of door belonging to species to be measured.The method of the present invention only needs the ITS2 gene orders of testing sample or doubtful species and target species carrying out degree of ramification comparison, more simple and easy to do, accuracy is high, and objectivity is strong, it need not largely be sampled as reference to belonging to other species together, there is higher application value.
Description
Technical field
The present invention relates to technical field of biological, more particularly to a kind of authentication method and purposes of seed plant species.
Background technology
The main method of species identification is morphological classification method at present, however because of its poor accuracy, subjectivity is strong, efficiency
It is low, the requirement that today, people were identified species taxonomy can not have been met.With the horizontal raising of sequencing and it is sequenced the reduction of cost,
Effect of the DNA sequence dna in species identification is more and more prominent.Canadian zoologist Paul Hebert are proposed first within 2003
DNA barcoding (DNA bar code) concept.It with it is a segment standard, having enough variations, easily amplification and it is relatively short
DNA fragmentation, by establishing biological DNA bar code database, realize quick, accurate and automation species identification.DNA bar code
Technology is that current species identification uses most wide technology, however as going deep into for research, the side based on DNA bar code identification species
Method is increasingly denounced, and is primarily due to species coverage and standard that DNA bar code technology depends critically upon reference database
True property.When database is not complete, sample to be tested is often reflected because the master sample with database does not compare by mistake
It is fixed.Because species and its sibling species are in often worldwide distribution, most of species and its sibling species are difficult that sampling is complete, this into
For the bottleneck of limitation DNA bar code technology application.
Known genetic distance is the index of some character complex inheritance difference sizes between measurement kind.Required by breeding objective
Character more than one, in order to more fully reflect the hereditary difference between parental breed, it is necessary to multiple characters synthesis examine
Consider, so as to the concept for genetic distance of extending out.Because the various characters of biology have gene to express, lost in molecular biology
The difference degree that distance refers to gene between different plant species or the different colony of same species is passed, the method for estimation mainly has JC69
Method, Kimura two parameter methods（K2P methods）, distance coefficient method, pivot coacervation and principal component analysis etc., the wherein application of K2P methods the most
Widely.In the past similar research does not consider species Genetic distance numerical value when identifying species, or it is determined that species inter-species
During genetic distance, averaged only by belonging to interior species together and comparing two-by-two to obtain, its value is substantially bigger than normal.
As can be seen here, deficiency of the prior art can be based on, there is provided a kind of authentication method and use of seed plant species
On the way, can it is quick, simple and it is accurate identification seed plant species classification, turn into those skilled in the art's technology urgently to be resolved hurrily
Problem.
The content of the invention
The present invention is directed to above-mentioned technical problem, proposes a kind of authentication method and purposes of seed plant species, passes through kind
The ITS2 gene order big datas of sub- plant species, analyze the genetic distance between substantial amounts of sibling species, its heredity obtained
Distance values are relatively stable in different taxonomic category or monoid, and point of seed plant species identification is used as by calculating average value
Sub- dividing value, species to be measured can comparatively fast be identified, substantially increase efficiency and the degree of accuracy of identification.
A kind of authentication method of seed plant species, comprises the following steps：
Determine molecule dividing value：Selected seed plant species, the ITS2 gene orders of the seed plant species are obtained, based on ITS2
Gene order builds chadogram, and multipair sibling species are chosen in chadogram, calculates the genetic distance between each pair sibling species, to category
Significance test of difference is carried out in multigroup genetic distance of same door species, the inapparent genetic distance of difference is chosen and calculates averagely
It is worth to molecule dividing value；
Determine species classification：The genetic distance between species and target species to be measured is calculated, by the genetic distance and species to be measured
The molecule dividing value of affiliated door compares, and as it is more than molecule dividing value, then species to be measured are not belonging to target species, as it is less than molecule circle
Value, then determinand kind belongs to target species.
As preferred：The molecule dividing value of Angiospermae is 0.03361 ~ 0.04589；The molecule dividing value of Gymnospermae is
0.01178~0.02715。
As preferred：The molecule dividing value of the Angiospermae is 0.03975；The molecule dividing value of the Gymnospermae is
0.01946。
As preferred：Chadogram is built using adjacent method.
As preferred：ITS2 gene order matrixes are built to obtain chadogram in units of category, fetch bit is in the same of chadogram
Two neighboring species are as a pair of sibling species in one branch.
As preferred：The genetic distance of each sibling species is calculated using the K2P models in MEGA softwares.
As preferred：The genetic distance of each pair sibling species is taken to carry out significance test of difference, selection is located at 95% confidential interval
Interior genetic distance simultaneously calculates average value.
As preferred：The method for calculating the genetic distance between species and target species to be measured comprises the following steps：Extraction is treated
Survey the ITS2 gene orders of species and be sequenced, by the ITS2 bases of target species in the ITS2 gene orders of testing sample and database
Because sequence is compared, the genetic distance between the species to be measured and target species is calculated.
As preferred：Specifically include following steps：
Determine molecule dividing value：In category more than 75% seed plant species are chosen, obtain the ITS2 genes of the seed plant species
Sequence, ITS2 gene order matrixes are built in units of category to obtain chadogram, fetch bit is in adjacent in the same branch of chadogram
Two species calculate the genetic distance of each sibling species, multiple genetic distances to belonging to same door species as a pair of sibling species
Significance test of difference is carried out, the calculating of the genetic distance in 95% confidential interval is chosen and is averagely worth to Angiospermae
Molecule dividing value is 0.03361 ~ 0.04589, and the molecule dividing value of Gymnospermae is 0.01178 ~ 0.02715；
Determine species classification：Extract the ITS2 gene orders of species to be measured and be sequenced, by the ITS2 gene orders of testing sample with
The ITS2 gene orders of target species are compared in database, calculate heredity between the species to be measured and target species away from
From by the genetic distance compared with the molecule dividing value of door belonging to species to be measured, as it is more than molecule dividing value, then species to be measured are not
Belong to target species, as it is less than molecule dividing value, then determinand kind belongs to target species.
A kind of application in medicinal material or instruction plant in authentication of authentication method of above-mentioned seed plant species.
Compared with prior art, the advantages and positive effects of the present invention are：
1st, the authentication method of a kind of seed plant species provided by the invention, the sibling species genetic distance used are closest to species
The numerical value of breeding true difference, therefore the qualification result degree of accuracy is higher.Other the method for the invention is entered using molecule dividing value
The identification of row seed plant species, its coverage to database require low, it is only necessary to by testing sample or doubtful species with
The ITS2 gene orders of target species carry out degree of ramification comparison, and method of the invention is more simple and easy to do, and accuracy is high, visitor
The property seen is strong, it is not necessary to is largely sampled as reference to belonging to other species together, has higher application value；
2nd, the present invention is after selected seed plant species, it is contemplated that genetic diversity in species be present and inbred genetic diversity is small
Strictly to be got together in inter-species, therefore to a plurality of sequence of same species in evolution tree graph, it is former according to the structure of chadogram
Reason, the sibling species that two species got together are chosen have a highest gene similitude, i.e., nearest affiliation, therefore
Hereditary difference between the sibling species that the present invention obtains is higher than inbred genetic Diversity, is lost than belonging to interior any pair of inter-species together
Biography difference is low, and the molecule dividing value finally obtained is other species more than this value closest to the boundary of species hereditary difference；
3rd, in Chinese medicine inspection technology field, Chinese medicine in discrimination process there is being looked genuine with puppet, homonym and same
The complicated confusion phenomena such as thing different name, because the present invention is that the angle based on genetic distance goes to differentiate species, therefore pass through
It is more accurate that the method for the present invention carries out Materia Medica Identification.
Brief description of the drawings
Fig. 1 classifies for angiosperm APG II in a kind of authentication method embodiment of seed plant species of the present invention
Figure.
Embodiment
The technical scheme in the embodiment of the present invention will be clearly and completely described below, it is clear that described implementation
Example only part of the embodiment of the present invention, rather than whole embodiments.It is common based on the embodiment in the present invention, this area
The every other embodiment that technical staff is obtained under the premise of creative work is not made, belong to the model that the present invention protects
Enclose.
The embodiment of the present invention provides a kind of authentication method of seed plant species, comprises the following steps：
Determine molecule dividing value：Selected seed plant species, the ITS2 gene orders of the seed plant species are obtained, based on ITS2
Gene order builds chadogram, and multipair sibling species are chosen in chadogram, calculates the genetic distance between each pair sibling species, to category
Significance test of difference is carried out in multigroup genetic distance of same door species, the inapparent genetic distance of difference is chosen and calculates averagely
It is worth to molecule dividing value；
Determine species classification：The genetic distance between species and target species to be measured is calculated, by the genetic distance and species to be measured
The molecule dividing value of affiliated door compares, and as it is more than molecule dividing value, then species to be measured are not belonging to target species, as it is less than molecule circle
Value, then determinand kind belongs to target species.
By the ITS2 gene order big datas of seed plant species, analyze heredity between substantial amounts of sibling species away from
From its genetic distance numerical value obtained is relatively stable in different taxonomic category or monoid, is used as seed by calculating average value
Species to be measured can comparatively fast be identified, substantially increase efficiency and the degree of accuracy of identification by the molecule dividing value of plant species identification.
In an alternative embodiment：The molecule dividing value of Angiospermae is 0.03361 ~ 0.04589；Point of Gymnospermae
Sub- dividing value is 0.01178 ~ 0.02715.
Wherein the molecule dividing value of Angiospermae is the Gymnospermae 0.03361 ~ 0.04589 in 95% confidential interval
Molecule dividing value for 0.01178 ~ 0.02715 in 95% confidential interval.
In a preferred embodiment：The molecule dividing value of the Angiospermae is 0.03975；Point of the Gymnospermae
Sub- dividing value is 0.01946.
In an alternative embodiment：Chadogram is built using adjacent method.
In an alternative embodiment：ITS2 gene order matrixes are built to obtain chadogram in units of category, fetch bit is in entering
Two neighboring species are as a pair of sibling species in the same branch of change tree.
Evolved it should be noted that importing in MEGA softwares to build by ITS2 gene orders matrix in units of category
Tree, fetch bit is in two neighboring species in the same branch of chadogram as a pair of sibling species.
In an alternative embodiment：The genetic distance of each sibling species is calculated using the K2P models in MEGA softwares.
Wherein described K2P models full name is Kimura 2-parameter models.
In an alternative embodiment：Take the genetic distance of each pair sibling species to carry out significance test of difference, choose and be located at 95%
Genetic distance in confidential interval simultaneously calculates average value.
Significance test of difference is carried out it should be noted that the genetic distance of each pair sibling species is imported in SPSS softwares.
In an alternative embodiment：Calculating the method for the genetic distance between species and target species to be measured includes following step
Suddenly：Extract the ITS2 gene orders of species to be measured and be sequenced, by object in the ITS2 gene orders of testing sample and database
The ITS2 gene orders of kind are compared, and calculate the genetic distance between the species to be measured and target species.
It should be noted that the something lost between the species to be measured and target species is calculated based on the K2P models in MEGA softwares
Pass distance.
In a preferred embodiment：Specifically include following steps：
Determine molecule dividing value：In category more than 75% seed plant species are chosen, obtain the ITS2 genes of the seed plant species
Sequence, ITS2 gene order matrixes are built in units of category to obtain chadogram, fetch bit is in adjacent in the same branch of chadogram
Two species calculate the genetic distance of each sibling species, multiple genetic distances to belonging to same door species as a pair of sibling species
Significance test of difference is carried out, the calculating of the genetic distance in 95% confidential interval is chosen and is averagely worth to Angiospermae
Molecule dividing value is 0.03361 ~ 0.04589, and the molecule dividing value of Gymnospermae is 0.01178 ~ 0.02715；
Determine species classification：Extract the ITS2 gene orders of species to be measured and be sequenced, by the ITS2 gene orders of testing sample with
The ITS2 gene orders of target species are compared in database, calculate heredity between the species to be measured and target species away from
From by the genetic distance compared with the molecule dividing value of door belonging to species to be measured, as it is more than molecule dividing value, then species to be measured are not
Belong to target species, as it is less than molecule dividing value, then determinand kind belongs to target species.
It should be noted that the heredity between the species to be measured and target species is calculated based on the K2P models in MEGA softwares
Distance.
Another embodiment of the present invention provides a kind of authentication method of above-mentioned seed plant species and medicinal material or entered in authentication
The application invaded in plant.
In order to become apparent from introducing in detail a kind of authentication method for seed plant species that the embodiment of the present invention is provided and
Its purposes, is described below in conjunction with specific embodiment.
Embodiment 1
A kind of authentication method of seed plant species, comprises the following steps：
Determine molecule dividing value：According to as shown in Figure 1《Angiosperm APG II categorizing systems（Revised edition）》Shown monoid closes
It is and except monotype guiding principle（Ginkgopsida）All representative guiding principles of gymnosperm in addition, have chosen 7866 species, specifically comprising quilt
99 sections of plant, 179 category, 99 sections, make it substantially covers the diversity of angiosperm, the category where selected species
Meet more than 75% species coverage；The ITS2 gene orders of monoid are downloaded from GenBank, or download the ITS sequence of monoid
Interception is annotated according to GenBank and obtains ITS2 gene orders；By the ITS2 gene order matrixes of selected species in units of category
Import in MEGA softwares and build chadogram, a plurality of sequence of same species wants adjacent input, according to the principle of structure chadogram, choosing
Go out in chadogram has highest gene similar positioned at two adjacent species of same branch as sibling species, each pair of sisters' inter-species
Property, as shown in table 1, final 281 pairs of sibling species for obtaining 113 category.The genetic distance of 281 pairs of sibling species is imported into SPSS softwares
Middle carry out significance test of difference, calculate genetic distance greatest genetic distance, minimum genetic distance, genetic distance average value and something lost
Pass distance maximum and minimum ratio.Fetch bit is averagely worth to angiosperm in the genetic distance in 95% confidential interval and calculating
Molecule dividing value：0.03975,95% confidential interval is [0.03361,0.04589]；Molecule dividing value gymnospermous：0.01946,
95% confidential interval is [0.01178,0.02715].
The Genetic distance of 281 pairs of sibling species of 1 113 category of table
Determine species classification：A kind of authentication method of doubtful medicinal plant artemisia annua, specific implementation step include：
Step 1: species sequencing to be measured：DNA, PCR amplification of species to be measured are extracted, uses ITS universal primers ITS4 and ITS5B
As pcr amplification primer thing, PCR primer is subjected to DNA sequencing, the sequence results such as SEQ ID No of species to be measured：Shown in 1：
ITS4 sequence such as SEQ ID No：Shown in 5；ITS4 sequence such as SEQ ID No：Shown in 6；
Step 2: sequence alignment：Obtain target species artemisia annua respectively from GenBank（Artemisia annua）'s
GenBank Serial No. JQ230972, FJ528301, AY548199, KX581796 and GU724282 sequence, above-mentioned sequence number
Sequence such as SEQ ID No：Shown in 1, using CLUSTAL X softwares by the ITS2 gene orders of species to be measured and above-mentioned whole
Target species artemisia annua（Artemisia annua）ITS2 gene orders be compared；
Step 3: calculate genetic distance：The sequence matrix that will have been compared, MEGA softwares are imported, based on K2P models（Kimura 2-
parameter model）Genetic distance between the two is calculated, genetic distance value is 0.00446；
Step 4: identification：Genetic distance 0.00446 between doubtful medicinal plant artemisia annua and artemisia annua is less than angiosperm
Molecule dividing value 0.03975（95% confidential interval：0.03361-0.04589）, it is possible to identify it is artemisia annua.
Embodiment 2
Determine that molecule dividing value and the step described in embodiment 1 are consistent, therefore repeat no more in the present embodiment.
Determine species classification：A kind of authentication method of doubtful medicinal plant Chinese yew, specific implementation step include：
Step 1: species sequencing to be measured：DNA, PCR amplification of species to be measured are extracted, uses ITS universal primers ITS4 and ITS5B
As pcr amplification primer thing, PCR primer is subjected to DNA sequencing, the sequence results such as SEQ ID No of species to be measured：Shown in 4：
ITS4 sequence such as SEQ ID No：Shown in 5；ITS4 sequence such as SEQ ID No：Shown in 6；
Step 2: sequence alignment：Obtain target species Chinese yew respectively from GenBank（Taxus chinensis）'s
GenBank Serial No. AF259300, AF259292 and AF259291 sequence, wherein GenBank Serial No. AF259300
Sequence such as SEQ ID No：Shown in 2, GenBank Serial No. AF259292 and AF259291 sequence are such as SEQ ID No：
Shown in 3；Using CLUSTAL X softwares by the ITS2 gene orders of species to be measured and above-mentioned target complete species Chinese yew（Taxus chinensis）ITS2 gene orders be compared；
Step 3: calculate genetic distance：The sequence matrix that will have been compared, MEGA softwares are imported, based on K2P models（Kimura 2-
parameter model）Genetic distance between the two is calculated, genetic distance value is 0.00446；
Step 4: identification：Genetic distance 0.06012 between doubtful medicinal plant artemisia annua and artemisia annua is more than gymnospermous
Molecule dividing value 0.01946（95% confidential interval：0.01178-0.02715）, exclude the possibility that testing sample is Chinese yew.
Sequence table
<110>Shandong University
<120>The authentication method and purposes of seed plant species
<141> 2017-09-14
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 225
<212> DNA
<213>Artemisia annua (Artemisia annua)
<400> 1
cgcatcgcgt cgccccccac aattctctgt aaagggaact cgtgttttgg gggcggataa 60
tggtctcccg tgctcatggc gtggttggcc gaaataggag tcccttcgat ggacgcacga 120
actagtggtg gtcgtaaaaa ccctcgtctt ttgtttcgtg ccgttagtcg caagggaaac 180
tctaagaaaa ccccaacgtg tcgtctcttg acgacgcttc gaccg 225
<210> 2
<211> 233
<212> DNA
<213>Chinese yew (Taxus chinensis)
<400> 2
cactctaaaa tcgacctccc ccgtgctatg ggggaggatc ggagatggcc gtccgtgccc 60
accagcggtg cggttggctt aaacgagcac gggacttgtg gccaatgtca cgatgagcgg 120
tggcctgtgt gggtcggcgt tggattgtgg catgaggttg cggctttgtg tggaacttta 180
tttgtggttg gtcggagcct tccttcggga gggtccgtct ataactcttg cgg 233
<210> 3
<211> 233
<212> DNA
<213>Chinese yew (Taxus chinensis)
<400> 3
cactctaaaa tcgacttccc ccgttcctcg gcgggaggat cggagatggc tgtccgtgcc 60
caccagcggc gcggtcggct taaatgggca cgagatttgt ggccgatgtc acgatgagcg 120
gtggccaatg ttggtcggcg ttggattgtg gcatgaggcc acggatttgt gcggaacttt 180
actcgttgtt agtcgcgact cttccgcggg agggcgcgtc ccaactttcg cgg 233
<210> 4
<211> 233
<212> DNA
<213>Unknown (Unknown)
<400> 4
cactctaaaa tcgacttccc ccgttcctcg gcgggaggat cggagatggc tgtccgtgcc 60
caccagcggc gcggtcggct taaatgggca cgagatttgt ggccgatgtc acgatgagcg 120
gtggccaatg ttggtcggcg ttggattgtg gcatgaggcc acggatttgt gcggaacttt 180
actcgttgtt agtcgcgact cttccgcggg agggcgcgtc ccaactttcg cgg 233
<210> 5
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
tcctccgctt attgatatgc 20
<210> 6
<211> 22
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
ggaaggagaa gtcgtaacaa gg 22
Claims (9)
- A kind of 1. authentication method of seed plant species, it is characterised in that：Comprise the following steps：Determine molecule dividing value：Selected seed plant species, the ITS2 gene orders of the seed plant species are obtained, based on ITS2 Gene order builds chadogram, and multipair sibling species are chosen in chadogram, calculates the genetic distance between each pair sibling species, to category Significance test of difference is carried out in multigroup genetic distance of same door species, the inapparent genetic distance of difference is chosen and calculates averagely It is worth to molecule dividing value；Determine species classification：The genetic distance between species and target species to be measured is calculated, by the genetic distance and species to be measured The molecule dividing value of affiliated door compares, and as it is more than molecule dividing value, then species to be measured are not belonging to target species, as it is less than molecule circle Value, then determinand kind belongs to target species.
- 2. the authentication method of seed plant species according to claim 1, it is characterised in that：Molecule circle of Angiospermae It is worth for 0.03361 ~ 0.04589；The molecule dividing value of Gymnospermae is 0.01178 ~ 0.02715.
- 3. the authentication method of seed plant species according to claim 2, it is characterised in that：Point of the Angiospermae Sub- dividing value is 0.03975；The molecule dividing value of the Gymnospermae is 0.01946.
- 4. the authentication method of seed plant species according to claim 1, it is characterised in that：Built and evolved using adjacent method Tree.
- 5. the authentication method of seed plant species according to claim 4, it is characterised in that：In units of category by described in ITS2 gene order matrixes build to obtain chadogram, and fetch bit is in two neighboring species in the same branch of chadogram as a pair of elder sisters Younger sister's kind.
- 6. the authentication method of seed plant species according to claim 1, it is characterised in that：Take the heredity of each pair sibling species Distance carries out significance test of difference, chooses the genetic distance in 95% confidential interval and calculates average value.
- 7. the authentication method of the seed plant species according to any one of claim 1 ~ 6, it is characterised in that：Calculate determinand Kind and the method for the genetic distance between target species comprise the following steps：Extract the ITS2 gene orders of species to be measured and be sequenced, The ITS2 gene orders of testing sample are compared with the ITS2 gene orders of target species in database, calculated described to be measured Genetic distance between species and target species.
- 8. the authentication method of seed plant species according to claim 1, it is characterised in that：Specifically include following steps：Determine molecule dividing value：In category more than 75% seed plant species are chosen, obtain the ITS2 genes of the seed plant species Sequence, ITS2 gene order matrixes are built in units of category to obtain chadogram, fetch bit is in adjacent in the same branch of chadogram Two species calculate the genetic distance of each sibling species, multiple genetic distances to belonging to same door species as a pair of sibling species Significance test of difference is carried out, the calculating of the genetic distance in 95% confidential interval is chosen and is averagely worth to Angiospermae Molecule dividing value is 0.03361 ~ 0.04589, and the molecule dividing value of Gymnospermae is 0.01178 ~ 0.02715；Determine species classification：Extract the ITS2 gene orders of species to be measured and be sequenced, by the ITS2 gene orders of testing sample with The ITS2 gene orders of target species are compared in database, calculate heredity between the species to be measured and target species away from From by the genetic distance compared with the molecule dividing value of door belonging to species to be measured, as it is more than molecule dividing value, then species to be measured are not Belong to target species, as it is less than molecule dividing value, then determinand kind belongs to target species.
- 9. a kind of authentication method of the seed plant species described in any one of claim 1 ~ 8 medicinal material or instruction plant in authentication In application.
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| CN110111847A (en) * | 2019-04-24 | 2019-08-09 | 华中科技大学鄂州工业技术研究院 | Method and apparatus based on ITS2 plant identification species |
| CN110910959A (en) * | 2019-11-04 | 2020-03-24 | 中国水稻研究所 | Population genetic evolution map and construction method thereof |
| CN113160893A (en) * | 2021-06-09 | 2021-07-23 | 中国科学院昆明植物研究所 | Mining plant ITSs sequence from second generation sequencing data and using the same for identifying variety families |
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| CN113160893A (en) * | 2021-06-09 | 2021-07-23 | 中国科学院昆明植物研究所 | Mining plant ITSs sequence from second generation sequencing data and using the same for identifying variety families |
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