CN107164525A - A kind of DNA for differentiating 6 kinds of Pterocarpus timber combines bar code and its discrimination method and application - Google Patents
A kind of DNA for differentiating 6 kinds of Pterocarpus timber combines bar code and its discrimination method and application Download PDFInfo
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- CN107164525A CN107164525A CN201710511802.2A CN201710511802A CN107164525A CN 107164525 A CN107164525 A CN 107164525A CN 201710511802 A CN201710511802 A CN 201710511802A CN 107164525 A CN107164525 A CN 107164525A
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Abstract
Bar code is combined the invention discloses a kind of DNA for differentiating 6 kinds of Pterocarpus timber, and the DNA combinations bar code is any two or more or DNA combination bar codes that all combination is formed in bar code sequence matK, ndhF rpl32, rbcL or ITS2.The invention also discloses a kind of method and its application for differentiating 6 kinds of Pterocarpus timber.The DNA combination bar codes and its method that the present invention is provided can realize the accurate discriminating of 6 kinds of Pterocarpus timber, solving traditional timber recognition methods can not differentiate Pterocarpus timber to the problem of " kind " level, it is that the timber-trade law enforcement agencies such as customs, quality inspection quarantine and wood management trade personnel provide a kind of discrimination method of high efficient and reliable, with higher practical value.
Description
Technical field
The invention belongs to the molecular Biological Detection field of wood material species identification, and in particular to one kind differentiates 6 kinds of Pterocarpuses
The DNA combination bar codes of timber and its discrimination method and application.
Background technology
Pterocarpus (Pterocarpus Jacq.) is subordinate to pulse family (Leguminosae), about 70 kinds, is distributed mainly on the torrid zone
And subtropical zone.Pterocarpus timber is well-known throughout the world with its wood quality and medical value.Due to depositing for tremendous economic interests chain
Its demand is continuously increased with society, illegal and exhaustive exploitation makes Pterocarpus timber resources increasingly deficient.Therefore, red sandalwood
Category timber resources has gradually obtained the concern of International Lawful And Legal.At present, 2 seeds of red sandalwood and hedgehog red sandalwood, which are put into, is on the point of
Wild animals and plants kind of endangering international trade protection pact (CITES) annex II；Several Pterocarpus are also put into international guarantor naturally
Protect alliance (IUCN) Red List；Meanwhile, India red sandalwood was included in national key protected wild plants in 1999 by Chinese Government
Register.
Pterocarpus timber varieties of trees is identified as protecting the implementation of timber resources and laws and regulations to provide technical support.However,
Conventional identification techniques based on the anatomy of wood can only differentiate Pterocarpus timber to " category " or " class ", it is impossible to realize " kind " level
Discriminating.And emerging DNA bar code technology provides possibility for the identification of Pterocarpus timber " kind " level.
DNA bar code be a segment standard, with enough variations, easily amplification and relatively short DNA fragmentation.In recent years
Come, DNA bar code has been developed as the important tool of species identification.But unique DNA bar code is for species especially for entering
The recognition capability of change relation complexity species is relatively limited.And DNA combination bar codes have more by the combination of a plurality of unique sequence
Abundant sequence information site, and then species identification ability is improved, it is a kind of effective species discrimination method.
On the whole, Pterocarpus wood species are enriched, and the conventional identification techniques based on the anatomy of wood are difficult to timber
The identification of " kind " level, and emerging DNA bar code technology, particularly DNA combination bar code progressively develop into timber identification
Effective tool.
The content of the invention
The technical problem to be solved in the present invention is can not accurately to differentiate 6 kinds of Pterocarpus wood for traditional timber identification technology
Material.
So, the first object of the present invention is to provide a kind of DNA combination bar codes for differentiating 6 kinds of Pterocarpus timber.
The second object of the present invention is to provide a kind of method for differentiating 6 kinds of Pterocarpus timber.
The third object of the present invention is to provide the DNA combination bar codes and molecular biology method is differentiating 6 kinds of red sandalwoods
Belong to the application in timber.
Based on this, technical scheme is as follows：
Bar code is combined present invention firstly provides a kind of DNA for differentiating 6 kinds of Pterocarpus timber, the DNA combines bar shaped
Code is any two or more or DNA groups that all combination is formed in bar code sequence matK, ndhF-rpl32, rbcL or ITS2
Close bar code.
It is preferred that, the DNA combinations bar code is matK+ndhF-rpl32+ITS2 combination bar codes.The DNA combobars
Shape code is that cell nucleus gene ITS2 sequences are combined with plastogene (matK and ndhF-rpl32).The matK+ndhF-rpl32+
ITS2 sequences are as shown in SEQ ID No.9-14.
It is preferred that, 6 kinds of Pterocarpus timber is respectively kiaat Pterocarpus angolensis DC., print
Spend red sandalwood Pterocarpus indicus Willd., Burma padauk Pterocarpus macrocarpus Kurz, santal purple
Wingceltis Pterocarpus santalinus L.f., African padank Pterocarpus soyauxii Taub. and dyestuff red sandalwood
Pterocarpus tinctorius Welw.。
Further, the invention provides a kind of method for differentiating 6 kinds of Pterocarpus timber, this method utilizes DNA combobars
Shape code, and differentiate by molecular biology method 6 kinds of Pterocarpus timber；The DNA combinations bar code is bar code sequence
In matK, ndhF-rpl32, rbcL or ITS2 it is any two or more or all combination formed DNA combination bar code.
Specifically, it the described method comprises the following steps：
(1) by wood sample milled processed into wood powder；
(2) DNA of the wood powder sample obtained by extraction step (1)；
(3) obtained DNA is extracted as template using step (2), PCR expands matK, ndhF-rpl32 and ITS2 bar shaped
Code sequence；
(4) pcr amplification product for obtaining step (3) is sequenced, and obtains matK, ndhF-rpl32 and ITS2 sequence,
And it is combined；
(5) TaxonDNA methods and system are based on the matK+ndhF-rpl32+ITS2 bar codes combination obtained by step (4)
Develop chadogram method and differentiate 6 kinds of Pterocarpus timber.
It is preferred that, the wood powder described in step (1) be ground to 200 mesh and more than.
It is preferred that, DNA need to be purified described in step (2), the final concentration of 1-200ng/ μ L of DNA.
It is preferred that, the matK primer sequences of step (3) the PCR amplifications are as shown in SEQ ID No.1-2, ndhF-rpl32
Primer sequence is as shown in SEQ ID No.3-4, and ITS2 primer sequences are as shown in SEQ ID No.5-6.
It is preferred that, step (3) the pcr amplification reaction system is 0.5-3U archaeal dna polymerases, 1.0-2.0mM MgCl2、
200 μM of single dNTP, 0.1-5.0 μM of single primers, pH7.0-8.0 0.1-1.5mg/mL bovine serum albumin(BSA)s and 10-
1000ng template DNAs；PCR reaction conditions are 94-96 DEG C of pre-degeneration 0.5-10min；94-96 DEG C of denaturation 0.05-2min, 40-65
DEG C annealing 0.5-2min, 72 DEG C extension 0.3-2min, circulate 20-50 times；72 DEG C extend 2-15min eventually.
It is preferred that, TaxonDNA methods are based on " best match " and " 2 indexs of best close match " in step (5)
Differentiate；Phylogenetic evolution tree method is differentiated based on adjacent method.
Further, the invention provides the side of above-mentioned DNA combination bar codes, or 6 kinds of Pterocarpus timber of above-mentioned discriminating
Application of the method in the discriminating of Pterocarpus timber.
The invention has the advantages that：
1st, the present invention combines bar code based on a kind of DNA, it is possible to achieve the accurate discriminating of 6 kinds of Pterocarpus timber, breaches
Limitation of the traditional timber identification technology None- identified timber to " kind "；
2nd, the diagnostic primerses specificity that the present invention is screened is good, and amplification and sequencing success rate are higher；
3rd, currently preferred matK+ndhF-rpl32+ITS2 combined sequences DNA bar code is in 6 kinds of timber of Pterocarpus
In the presence of obvious difference site, with very strong distinguishing ability；
4th, currently preferred TaxonDNA methods and phylogenetic evolution tree method are simple to operate, and comprehensive based on both approaches
Close and distinguish seeds, recognition accuracy is high；
5th, sampling amount of the present invention is few, and not by materials position (sapwood, heart sapwood transition region and heartwood), the original shape of sample
State (wooden unit, wood powder) factor influences；
6th, experiment condition of the present invention relies on few, and general PCR Lab can be met；
7th, the present invention accurately identifying there is provided a kind of new approach and thinking for Pterocarpus timber.
In a word, the DNA combination bar codes and its method that the present invention is provided can realize the accurate mirror of 6 kinds of Pterocarpus timber
Not, Pterocarpus timber can not be differentiated to the problem of " kind " level by solving traditional timber recognition methods, be customs, quality inspection inspection
Yi Deng timber-trades law enforcement agency and wood management trade personnel provide a kind of discrimination method of high efficient and reliable, with higher reality
With value.
Brief description of the drawings
" best match " (A) and " best close match " (B) single and combination of the Fig. 1 based on TaxonDNA methods
The species identification success rate of bar code；
Fig. 2 sets (A) matK+ndhF-rpl32+ITS2, (B) based on the DNA systematic growth adjoinings for combining bar code structure
matK+rbcL+ITS2；
Fig. 3 combines bar code matK+ndhF-rpl32+ITS2 based on DNA and differentiates that the systematic growth of unknown sample abuts tree.
Embodiment
Following examples are merely to illustrate the present invention, but are not limited to the invention scope of the present invention.The technical field
Technician can make the modifications and adaptations of some non-intrinsically safes according to the content of foregoing invention.
Embodiment 1：Pterocarpus timber specific primer design
(1) plastid DNA sequence matK, ndhF-rpl32, rbcL and nucleus DNA of Pterocarpus are downloaded from GenBank
Sequence ITS2 (table 1)；
The Pterocarpus DNA bar code sequence downloaded in the GenBank of table 1
(2) Clustal X1.81 software aligned sequences are applied, and searches and determines 4 kinds of DNA bar code sequences between Pterocarpus
Difference site；
(3) softwares of application Primer Premier 5 are to tetra- barcode designs of matK, ndhF-rpl32, rbcL and ITS2
Primer.The amplimer is as shown in table 2.
2 four kinds of DNA bar code primer information of table
Embodiment 2：The DNA combinations bar code of 6 kinds of Pterocarpus standard of wood samples preferably with determination
(1) standard sample collection.
6 kinds of Pterocarpus standard samples (are respectively kiaat, India red sandalwood, Burma padauk, red sandalwood, Africa purple
Wingceltis and dyestuff red sandalwood) it is total No. 39, it is taken from China Forestry Science Research Institute's wood herbarium.
(2) sample preparation.
Wood sample is chosen, is cut off wood sample outer surface using the scalpel blade after 70% alcohol disinfecting, to avoid
External source pollutes；Wood sample is cut into some wood chips, low temperature precooling 3min in cryogenic freezing beveller is placed in, 3min is ground,
Running frequency 10cps；After grinding terminates, cross in 200 eye mesh screens, the microcentrifugal tube that thin wood powder is dispensed into some 50mL, often
Pipe wood powder amount 500mg, is placed in -80 DEG C of low temperature refrigerators and saves backup.
(3) DNA is extracted.
In the ultra-clean working environment disinfected, with reference to article (Jiao L, Yin Y, Cheng such as bibliography Jiao
Y,Jiang X.DNA barcoding for identification of the endangered species
Aquilaria sinensis:comparison of data from heated or aged wood
samples.Holzforschung.2014；68(4):DNA extractions 487-494.) are carried out to 6 kinds of Pterocarpus timber.
(4) pcr amplification reaction and sequencing.
Primer pair is four kinds of DNA bar code aligning primers pair in embodiment 1, and performing PCR expansion is entered by template of DNA extracts
Increase, reaction system is 30 μ L：The wherein μ L of Premix Ex Taq 15 (including l.25U Ex Taq archaeal dna polymerases, 2mM
MgCl2, 200 μM of single dNTP), 0.2 μM of single primer and about 20ng template DNAs.All PCR reactions are in PCR amplification instrument
Carry out.
Response procedures are：94 DEG C of pre-degeneration 2min；94 DEG C of denaturation 15s, 52 DEG C (matK), 47 DEG C of (ndhF-rpl32), 50
DEG C (rbcL) and 51 DEG C (ITS2) annealing 30s, 72 DEG C of extension 20s, are circulated 40 times；72 DEG C extend 7min eventually, you can obtain efficient
The DNA purpose fragments of amplification.Amplified production is subjected to two-way direct Sequencing after purification.
(5) sequence alignment analysis and combined sequence.
Sequencing quality assessment is carried out to sequencing result using ContigExpress softwares, two ends low quality part is removed, and
Quality evaluation is carried out to remainder, if meeting quality requirement, sequence assembly and check and correction can be used for.6 kinds of Pterocarpus timber
Tetra- kinds of bar code sequence GenBank accession number of matK, ndhF-rpl32, rbcL and ITS2 and sequence signature information respectively such as table
3 and table 4 shown in.
Four kinds of bar codes of 6 kinds of Pterocarpus wood species are combined respectively using Editseq softwares, matK+ is obtained
ITS2、matK+ndhF-rpl32、matK+rbcL、ndhF-rpl32+ITS2、ndhF-rpl32+rbcL、rbcL+ITS2、matK
+ndhF-rpl32+ITS2、matK+ndhF-rpl32+rbcL、matK+rbcL+ITS2、ndhF-rpl32+rbcL+ITS2、
11 DNA bar code combinations such as matK+ndhF-rpl32+rbcL+ITS2.
Four kinds of bar code sequence GenBank accession number of 6 kinds of Pterocarpus timber that 3 research sequencings of table are obtained
4 four kinds of DNA bar code sequence signature information of table
(6) combined sequence DNA bar code preferably with determination.
Distance and inter-species distance in the kind of bar code are combined to single and DNA by K2P distance models using MEGA softwares
Calculating analysis is carried out, it is specific as shown in table 5.
Utilize " best match " and " 2 indexs discriminating seeds of best close match ", knot in TaxonDNA softwares
Fruit shows that matK+ITS2 and matK+ndhF-rpl32+ITS2 can recognize 6 kinds of Pterocarpus timber varieties of trees (Fig. 1) of the above；Using
MEGA softwares are based on adjacent method phylogenetic tree construction, and matK+ndhF-rpl32+ITS2 and matK+rbcL+ITS2 are to 6 kinds of purples
Wingceltis belongs to the species identification success rate (Fig. 2) that timber has 100%.
Two methods of TaxonDNA methods and systematic evolution tree method are comprehensively utilized, single bar code and 11 bar codes are combined
Screened, it is determined that the optimal DNA combination bar codes of 6 kinds of Pterocarpus timber of identification are matK+ndhF-rpl32+ITS2 (SEQ
ID No.9-14)。
Table 5 is single and DNA is combined in the kind of bar code and inter-species distance
Embodiment 3：Unknown wood sample DNA differentiates
(1) 1 piece of wood sample is derived from lumber market, by carrying out anatomic construction identification to it, is accredited as narra class wood
Material, but red sandalwood seeds can not be judged.Therefore timber is differentiated based on DNA methods.
(2) prepare wood powder and be ground to 200 mesh and more than.
(3) timber DNA is extracted.
In the ultra-clean working environment disinfected, with reference to the DNA extraction method in embodiment 2, DNA is carried out to timber and carried
Take.The DNA also needs to be purified, the final concentration of 1-200ng/ μ L of DNA.
(4) pcr amplification reaction and sequencing.
Timber DNA extracts are that template enters performing PCR amplification, and amplimer is as shown in SEQ ID 1-6.Reaction system is 30 μ
L：The wherein μ L of Premix Ex Taq 15 (including l.25U Ex Taq archaeal dna polymerases, 2mM MgCl2, 200 μM of single dNTP),
0.2 μM of single primer and about 20ng template DNAs.All PCR reactions are carried out in PCR amplification instrument.Response procedures are：94 DEG C pre-
It is denatured 2min；94 DEG C of denaturation 15s, 52 DEG C (matK), 47 DEG C (ndhF-rpl32) and 51 DEG C (ITS2) anneals 30s, 72 DEG C respectively
Extend 20s, circulate 40 times；72 DEG C extend 7min eventually, you can obtain the DNA purpose fragments of efficient amplification.Amplified production is purified
After carry out two-way direct Sequencing.
(5) sequence alignment analysis differentiates with seeds.
The sequencing result of tri- sequences of matK, ndhF-rpl32 and ITS2 is combined, using composite sequence matK+
NdhF-rpl32+ITS2 constructing systems develop chadogram, as shown in Figure 3.Analysis result shows that the sample can be with red sandalwood
Sample clustering, and distinguished with other Pterocarpus.Cluster result proves that the sample is red sandalwood.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
SEQUENCE LISTING
<110>Chinese Academy Of Forestry Research Institute Of Wood Industry
<120>A kind of DNA for differentiating 6 kinds of Pterocarpus timber combines bar code and its discrimination method and application
<130> 011701703
<160> 14
<170> PatentIn version 3.5
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gggtgggagg ggtgaatgct ggcttcccgt gagcgagtgc ctcacggttg gctgaaaatc 600
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tctatttctg gcaatgtcat tttgatgttt ggtctcaacc aggaacgatc catataaacc 180
aattattatc cgagcattca tttaactttt tttggggggg ttatctttca aatgtgcggt 240
tcgtaatgag ttttgtttct tattcgtcgg ttttatctct tttgtaaagg cttcagttaa 300
taaaaaagag tgaacatata aatagaattt tctattattc tgaatctctg cacaatactt 360
ctaatattgc aaagtacaaa gttaaaatgg tccaataaac aaattcgtgt gaattgcaga 420
atcccgtgaa ccatcgagtc tttgaacgca agttgcgccc gaagccatta ggctaagggc 480
acgcctgcct gggtgtcaca aatcgttgcc ccaatccccg cgccgctcct gtgggcgccg 540
ggctgcgggg cgaatgctgg cttcccgtga gcgagtgcct cgcggttggc cgaaaatcgg 600
gttcgtggtg gagggtaagc gccatgacag acggtggttg agcgcaatct cgaggccagt 660
cgtgcgcggt cccctccgct agttacggac tccgtgaccc gtgagcggca ccgatcgccc 720
atgatgcgac ctcaggtcag gcggggcta 749
<210> 13
<211> 748
<212> DNA
<213>African padank composite sequence
<400> 13
tttcgtctac cttatccttc ttcaaggatc ctttgattca ttatgttaga tatcaaggaa 60
aatccattct ggcttcaaag aatgcgcctc ttttgatgaa taaatggaaa tactatctca 120
tctatttctg gcaatgtcat tttgatgttt ggtctcaacc aggaacgatc catataaacc 180
aattattatc cgagcattca tttaactttt tttggggggg ctatctttca aatgtgcggt 240
tcgtaatgag ttttgtttct tattcgtcgg ttttatctct tttgtaaagg cttcagttaa 300
taaaaaagag tgaacatata aatagaattt tctattattc tgaatctctg cacaatactt 360
ctaatattgc aaagtacaaa gttaaaatgg tccaataaac aaattcgtgt gaattgcaga 420
atcccgtgaa ccatcgagtc tttgaacgca agttgcgccc gaagccatta ggctaagggc 480
acgcctgcct gggtgtcaca aatcgttgcc ccaatcaatc ccggcgccta ataaaggcgc 540
tgggcggggc gaatgctggc ttcccgtgag cgagtgcctc gcggctggcc gaaaatcggg 600
tccgtggtgg agggcagcgc catgacagac ggtggttgag cgcaatctcg aggccagtcg 660
tgcgcggtcc cctctgctag ctacggactc cgtgacccgt ggagcggcac cgatcgccca 720
tgatgcgacc tcaggtcagg cggggcta 748
<210> 14
<211> 751
<212> DNA
<213>Dyestuff red sandalwood composite sequence
<400> 14
tttcgtctac cttatccttc ttcaaggatc ctttgattca ttatgttaga tatcaaggaa 60
aatccattct ggcttcaaag aatgcgcctc ttttgatgaa taaatggaaa tactatctca 120
tctatttctg gcaatgtcat tttgatgttt ggtctcaacc aggaacgatc catataaacc 180
aattattatc cgagcattca tttaactttt tttggggggg ctatctttca aatgtgcggt 240
tcgtaatgag ttttgtttct tattcgtcgg ttttatctct tttgtaaagt cttcagttaa 300
taataataaa aaagagtgaa catataaata gaattttcta ttattctgaa tctctgcaca 360
atacttctaa tattgcaaag tacaaagtaa aaatggtcca ataaacaaat tcgtgtgaat 420
tgcagaatcc cgtgaaccat cgagtctttg aacgcaagtt gcgcccgaag ccattaggct 480
gagggcacgc ctgcctgggt gtcaccaatc gttgccccaa ccccagcgcc tctactctag 540
ggcactgggc ggggtgaatg ctggcttccc gtgagcgagt gcctcgcggt tggccgaaaa 600
tcgggttcgt ggtggagggt agcgccatga cagacggtgg ttgagtgcaa tctcgaggcc 660
agtcgtgcgc gtcccctccg ccggttacgg actccgtgac ccgtgagcgg caccgatcgc 720
ccatgatgcg acctcaggtc aggcggggct a 751
Claims (10)
1. a kind of DNA combination bar codes for differentiating 6 kinds of Pterocarpus timber, it is characterised in that the DNA combinations bar code is bar shaped
Any two or more or DNA combination bar codes that all combination is formed in code sequence matK, ndhF-rpl32, rbcL or ITS2.
2. DNA as claimed in claim 1 combines bar code, it is characterised in that the DNA combinations bar code is matK+ndhF-
Rpl32+ITS2 combines bar code.
3. DNA as claimed in claim 2 combines bar code, it is characterised in that the matK+ndhF-rpl32+ITS2 sequences
As shown in SEQ ID No.9-14.
4. the DNA combination bar codes as described in claims 1 to 3 any one, it is characterised in that 6 kinds of Pterocarpus timber
Respectively kiaat Pterocarpus angolensis DC., India red sandalwood Pterocarpus indicus
Willd., Burma padauk Pterocarpus macrocarpus Kurz, red sandalwood Pterocarpus santalinus
L.f., African padank Pterocarpus soyauxii Taub. and dyestuff red sandalwood Pterocarpus tinctorius
Welw.。
5. a kind of method for differentiating 6 kinds of Pterocarpus timber, it is characterised in that this method combines bar code using DNA, and by dividing
Sub- biological means differentiate 6 kinds of Pterocarpus timber；DNA combination bar code be bar code sequence matK, ndhF-rpl32,
In rbcL or ITS2 it is any two or more or all combination formed DNA combination bar code.
6. method as claimed in claim 5, it is characterised in that comprise the following steps：
(1) by wood sample milled processed into wood powder；
(2) DNA of the wood powder sample obtained by extraction step (1)；
(3) obtained DNA is extracted as template using step (2), PCR expands matK, ndhF-rpl32 and ITS2 bar code sequence
Row；
(4) pcr amplification product for obtaining step (3) is sequenced, and obtains matK, ndhF-rpl32 and ITS2 sequence, and right
It is combined；
(5) TaxonDNA methods and systematic growth are based on the matK+ndhF-rpl32+ITS2 bar codes combination obtained by step (4)
Chadogram method differentiates 6 kinds of Pterocarpus timber.
7. method as claimed in claim 6, it is characterised in that the positive anti-primer such as SEQ ID No.1-2 of amplification matK sequences
It is shown, the positive anti-primer such as SEQ ID No.3-4 of ndhF-rpl32 sequences are expanded, the positive anti-primer such as SEQ of ITS2 sequences is expanded
Shown in ID No.5-6.
8. method as claimed in claim 6, it is characterised in that step (3) the pcr amplification reaction system is 0.5-3U DNA
Polymerase, 1.0-2.0mM MgCl2, 200 μM of single dNTP, 0.1-5.0 μM of single primer, pH7.0-8.0 0.1-1.5mg/
ML bovine serum albumin(BSA)s and 10-1000ng template DNAs；PCR reaction conditions are 94-96 DEG C of pre-degeneration 0.5-10min；94-96℃
0.05-2min, 40-65 DEG C of annealing 0.5-2min, 72 DEG C of extension 0.3-2min are denatured, is circulated 20-50 times；72 DEG C extend 2- eventually
15min。
9. method as claimed in claim 6, it is characterised in that in step (5) TaxonDNA methods be based on " best match " and
" 2 indexs of best close match " differentiate；Phylogenetic evolution tree method is differentiated based on adjacent method.
10. DNA described in Claims 1 to 4 any one combines bar code, or differentiates 6 described in claim 5~9 any one
Plant application of the method for Pterocarpus timber in the discriminating of Pterocarpus timber.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108192897A (en) * | 2018-02-11 | 2018-06-22 | 云南省烟草农业科学研究院 | One grows tobacco rbcl genetic fragments and its application |
| CN110257543A (en) * | 2019-06-14 | 2019-09-20 | 浙江省检验检疫科学技术研究院 | Identify method, the primer and probe of Burma padauk |
| CN114540532A (en) * | 2022-03-04 | 2022-05-27 | 中国林业科学研究院木材工业研究所 | DNA bar code and method for identifying multiple wood in Machilus and Machilus |
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| CN106636412A (en) * | 2016-12-28 | 2017-05-10 | 广东产品质量监督检验研究院 | Method for recognizing Dalbergia and Pterocarpus by ITS (Internal Transcribed Spacer) sequence |
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| CN106636412A (en) * | 2016-12-28 | 2017-05-10 | 广东产品质量监督检验研究院 | Method for recognizing Dalbergia and Pterocarpus by ITS (Internal Transcribed Spacer) sequence |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108192897A (en) * | 2018-02-11 | 2018-06-22 | 云南省烟草农业科学研究院 | One grows tobacco rbcl genetic fragments and its application |
| CN110257543A (en) * | 2019-06-14 | 2019-09-20 | 浙江省检验检疫科学技术研究院 | Identify method, the primer and probe of Burma padauk |
| CN110257543B (en) * | 2019-06-14 | 2024-02-27 | 浙江省检验检疫科学技术研究院 | Method, primer and probe for identifying pterocarpus macrophylla |
| CN114540532A (en) * | 2022-03-04 | 2022-05-27 | 中国林业科学研究院木材工业研究所 | DNA bar code and method for identifying multiple wood in Machilus and Machilus |
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| Publication number | Publication date |
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| CN107164525B (en) | 2018-07-06 |
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