CN105838806A - Standard gene for identifying pterocarpus santalinus molecules and molecular identification method - Google Patents
Standard gene for identifying pterocarpus santalinus molecules and molecular identification method Download PDFInfo
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- CN105838806A CN105838806A CN201610326119.7A CN201610326119A CN105838806A CN 105838806 A CN105838806 A CN 105838806A CN 201610326119 A CN201610326119 A CN 201610326119A CN 105838806 A CN105838806 A CN 105838806A
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- red sandalwood
- pterocarpus santalinus
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- pterocarpus
- standard gene
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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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- 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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- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
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Abstract
Pterocarpus santalinus is also known as a red sandalwood and is an apapilionaceae pterocarpus plant. Because that material quality is good, the distribution area is small, a material forming cycle is long, and the market prices are soaring, pterocarpus santalinus counterfeits spread unchecked under the huge benefit drive, and how to accurately identify the pterocarpus santalinus becomes the biggest problem in the pterocarpus santalinus trading market. The invention provides a standard gene for identifying pterocarpus santalinus molecules and a molecular identification method. The method is applied to pterocarpus santalinus identification and has the advantages of being quick and accurate, bringing small sample damages and the like.
Description
Technical field
The invention belongs to biology field, be specifically related to standard gene and Molecular Identification side that plant molecular is identified
Method.
Background technology
Leaflet red sandalwood (Pterocarpus santalinus), has another name called red sandalwood, for Papilionaceae Pterocarpus plant.Little
Leaf red sandalwood is solid thick and heavy, wooden exquisiteness, and density is relatively big, and pinhole is less, and stability is excellent, and texture is beautiful, good toughness, resistance to chisels and carves, and produces
In South India and neighbouring narrow zone.Leaflet narra look is dark purple, is always the royal special timber of noble's furniture, price
Costliness, occupies first of each wood, is referred to as " wood of emperor ".18 the used furniture in th Century French imperial family Louis 18, its material is also
Being to select first-class leaflet red sandalwood, in the redwood seeds determined in " redwood GB ", leaflet red sandalwood is first of redwood.Leaflet red sandalwood
Growth is extremely slow, leaflet red sandalwood centuries to be grown the even more than one thousand years that becomes a useful person, and because volume recovery is extremely low, resource is in great shortage,
There is again saying of " very little wingceltis cun gold ".
Because material is excellent, distributed areas are little in addition, the cycle of becoming a useful person is long, and its market price constantly rises.In huge profit
Benefit is ordered about down, and the counterfeit of leaflet red sandalwood spreads unchecked, and how precise Identification leaflet red sandalwood becomes on leaflet red sandalwood trade market
Hang-up.The most mostly using microdissection and fluorescence observation method to identify leaflet red sandalwood, its operating process is complicated, damages sample
Wound is serious, and along with the progress of fraud technology, the qualification accuracy rate of these methods also can not be guaranteed.
Summary of the invention
For solving the problems referred to above, the present invention provides a kind of standard gene for leaflet red sandalwood Molecular Identification and Molecular Identification
Method, application the method identify leaflet red sandalwood, have quick, accurate, to features such as sample damage are little, concrete technical scheme is such as
Under.
(1) standard gene for leaflet red sandalwood Molecular Identification is obtained
According to DNA bar code know-why, use PCR amplification method, it is ensured that the purity of band and reliability.Sequencing result passes through
Manual check and correction, sequence assembly, and guarantee that gained sequence is standard sequence.From the leaflet red sandalwood sample extraction genomic DNA gathered,
With reference to plant bar code DNA fragmentation selection principle, utilize primer to carry out PCR (PCR), amplify sample corresponding
PsbK-psbI gene, post analysis comparison that amplified production is checked order, preferably primer sequence amplification leaflet red sandalwood sample psbK-
PsbI gene order fragment is as standard gene, and gene order is shown in sequence table (531 bases).
(2) method identifying leaflet red sandalwood by psbK-psbI gene order, is divided into four steps:
The first step, samples and extracts DNA；
Second step, utilizes PCR reaction amplification psbK-psbI gene order；
3rd step, checks order to amplified production；
4th step, carries out sequence analysis by the sequence obtained with the psbK-psbI gene order in claim 1, works as homology
Property base ratio less than 95% time, get rid of the possibility that these species are leaflet red sandalwood, when homology base ratio is more than or equal to 95%
Time, this sample be the probability of leaflet red sandalwood be × 100%.
Detailed description of the invention
In experiment, with the leaflet red sandalwood sample of 5 separate sources, extract sample by TIANGENE company DNA extraction kit
This DNA.The forward primer used: TTAGCCTTTGTTTGGCAAG, reverse primer: AGAGTTTGAGAGTAAGCAT, PCR expand
Condition is 94 DEG C of denaturations 4min, following 94 DEG C of sex change 45s, and 50 DEG C of annealing 45 s, 72 DEG C extend 1.5min, carry out altogether 35
Individual circulation, last 72 DEG C extend 10min.Obtaining product comparison after order-checking, the homology between sample is held at more than 99%,
Obtaining sequence with other species by online comparison, its homology is all below 52%.
Sequence table:
CTGCTGTAAGTTTTCGATGAGATCTTGAATTTGAATAAATGTCCTAAAAAAATTCGGAATTTATTCGAAAACA
AAAATTCGAACATTTTAACAATTTAAAAGATCAGATAAGTCTTACAGTGTGAATCCTCGATTCAAATATTGAAATTT
TTGGATAGACAAAAAAAATCTGGACCATCTCATTATTCCCTCATTCTTACGTTTTTTCCACTGAGAAATCCTTCTAT
TTTTAGATTTGTATTAGATTTGAGTCCACCACCAGTACCTGGGTTGTGGATATGAAAGAAAATCTTTTGTAATACAA
ATATTGGTAATAGTATATTGGTATTGGTAATACTATATTGGTAATAGTAATAAAGGGCTCGGCTCTTCCTACAAAGA
AAGAAATTTCCGAATTTTTTTTTCTATTTCCTTGAAATCACTTCATTTCTTAGAAACTTTGTATCAAAAGAAACATA
ATGTGTTATATAAGAGAATCTATTCTCTTTTCTCATTCTCTGTCGTTTTTTTTTTAATCTTGGAGATTTTGTA。
Sequence table:
CTGCTGTAAGTTTTCGATGAGATCTTGAATTTGAATAAATGTCCTAAAAAAATTCGGAATTTATTCGAAAACA
AAAATTCGAACATTTTAACAATTTAAAAGATCAGATAAGTCTTACAGTGTGAATCCTCGATTCAAATATTGAAATTT
TTGGATAGACAAAAAAAATCTGGACCATCTCATTATTCCCTCATTCTTACGTTTTTTCCACTGAGAAATCCTTCTAT
TTTTAGATTTGTATTAGATTTGAGTCCACCACCAGTACCTGGGTTGTGGATATGAAAGAAAATCTTTTGTAATACAA
ATATTGGTAATAGTATATTGGTATTGGTAATACTATATTGGTAATAGTAATAAAGGGCTCGGCTCTTCCTACAAAGA
AAGAAATTTCCGAATTTTTTTTTCTATTTCCTTGAAATCACTTCATTTCTTAGAAACTTTGTATCAAAAGAAACATA
ATGTGTTATATAAGAGAATCTATTCTCTTTTCTCATTCTCTGTCGTTTTTTTTTTAATCTTGGAGATTTTGTA
Claims (2)
1. the standard gene for leaflet red sandalwood Molecular Identification, it is characterised in that its amino acid sequence is the sequence in sequence table
Row.
2. the method identifying leaflet red sandalwood with the standard gene in claim 1, is divided into four steps:
The first step, samples and extracts DNA；
Second step, utilizes PCR reaction amplification psbK-psbI gene order, and front and back primer sequence is respectively as follows:
TTAGCCTTTGTTTGGCAAG、AGAGTTTGAGAGTAAGCAT；
3rd step, checks order to amplified production；
4th step, carries out sequence analysis by the sequence obtained with the psbK-psbI gene order in claim 1, works as homology
Property base ratio less than 95% time, get rid of the possibility that these species are leaflet red sandalwood, when homology base ratio is more than or equal to 95%
Time, this sample be the probability of leaflet red sandalwood be × 100%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610326119.7A CN105838806A (en) | 2016-05-17 | 2016-05-17 | Standard gene for identifying pterocarpus santalinus molecules and molecular identification method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610326119.7A CN105838806A (en) | 2016-05-17 | 2016-05-17 | Standard gene for identifying pterocarpus santalinus molecules and molecular identification method |
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| CN105838806A true CN105838806A (en) | 2016-08-10 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610326119.7A Pending CN105838806A (en) | 2016-05-17 | 2016-05-17 | Standard gene for identifying pterocarpus santalinus molecules and molecular identification method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106929575A (en) * | 2017-02-24 | 2017-07-07 | 中国林业科学研究院木材工业研究所 | A kind of miniature DNA bar code and its discrimination method and application for differentiating red sandalwood and dyestuff red sandalwood |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103820568A (en) * | 2014-03-20 | 2014-05-28 | 中国科学院上海有机化学研究所 | Special probe, primer, gene chip and method for detecting Pterocarpus indicus Wiild. |
-
2016
- 2016-05-17 CN CN201610326119.7A patent/CN105838806A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103820568A (en) * | 2014-03-20 | 2014-05-28 | 中国科学院上海有机化学研究所 | Special probe, primer, gene chip and method for detecting Pterocarpus indicus Wiild. |
Non-Patent Citations (2)
| Title |
|---|
| BENTE B. KLITGÅRD等: "A detailed investigation of the Pterocarpus clade (Leguminosae:Dalbergieae): Etaballia with radially symmetrical flowers is nested within the papilionoid-flowered Pterocarpus", 《SOUTH AFRICAN JOURNAL OF BOTANY》 * |
| 宁淑萍等: "植物DNA条形码研究进展", 《生物多样性》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106929575A (en) * | 2017-02-24 | 2017-07-07 | 中国林业科学研究院木材工业研究所 | A kind of miniature DNA bar code and its discrimination method and application for differentiating red sandalwood and dyestuff red sandalwood |
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