CN105925695A - Standard gene for molecular identification of pterocarpus macarocarpus kurz and molecular identification method - Google Patents

Standard gene for molecular identification of pterocarpus macarocarpus kurz and molecular identification method Download PDF

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Publication number
CN105925695A
CN105925695A CN201610326117.8A CN201610326117A CN105925695A CN 105925695 A CN105925695 A CN 105925695A CN 201610326117 A CN201610326117 A CN 201610326117A CN 105925695 A CN105925695 A CN 105925695A
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China
Prior art keywords
pterocarpus
macarocarpus
kurz
molecular identification
sequence
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CN201610326117.8A
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王兵益
杨时宇
廖礼彬
李体初
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Research Institute of Resource Insects of Chinese Academy of Forestry
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Research Institute of Resource Insects of Chinese Academy of Forestry
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Priority to CN201610326117.8A priority Critical patent/CN105925695A/en
Publication of CN105925695A publication Critical patent/CN105925695A/en
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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
    • 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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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Botany (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Pterocarpus macarocarpus kurz, which is hard in texture and high in density, is used for making high-grade furniture. Heartwood, which is slow in growth speed, needs a relatively long time in formation, market demand exceeds supply and market price is climbing. Due to temptation of huge benefit, counterfeits of the pterocarpus macarocarpus kurz spread everywhere, and it is the biggest problem for the transaction market of the pterocarpus macarocarpus kurz to accurately identify the pterocarpus macarocarpus kurz. The invention provides a standard gene for molecular identification of the pterocarpus macarocarpus kurz and a molecular identification method; and the method, when used for identifying the pterocarpus macarocarpus kurz, has the characteristics of being rapid, accurate, low in injury on a sample and the like.

Description

A kind of standard gene for Burma padauk Molecular Identification and method for identifying molecules
Technical field
The invention belongs to biology field, be specifically related to standard gene and method for identifying molecules that plant molecular is identified.
Background technology
Burma padauk (Pterocarpus macarocarpus Kurz), it is Papilionaceae Pterocarpus plant, mainly originates in Burma, Thailand and Laos.Burma padauk material is the heaviest, and density is high, and its heartwood is typically orange, brick red, aubergine, yellow or light yellow, often has dark striped；Outstanding feature is exactly strong fruital taste.It is generally used on building and joiner, noble furniture, timber floor, wheel, the deck of boat, implement handle and artware.
Because the heartwood speed of growth is slow, the time forming heartwood needs is longer, and supply falls short of demand in market, and its market price constantly rises.Under huge interests are ordered about, the counterfeit of Burma padauk spreads unchecked, and how precise Identification Burma padauk becomes the biggest problem on Burma padauk trade market.The most mostly using form observation to identify Burma padauk, its operating process is complicated, serious to sample damage, 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 Burma padauk Molecular Identification and method for identifying molecules, and application the method identifies Burma padauk, have quick, accurate, to features such as sample damage are little, concrete technical scheme is as follows.
(1) standard gene for Burma padauk Molecular Identification is obtained
According to DNA bar code know-why, use PCR amplification method.Sequencing result is by manual check and correction, sequence assembly, it is ensured that gained sequence is standard sequence.From the Burma padauk sample extraction genomic DNA gathered, with reference to plant bar code DNA fragmentation selection principle, utilize primer to carry out polymerase chain reaction (PCR), amplify sample correspondingpsbK-psbIGene, post analysis comparison that amplified production is checked order, preferably primer sequence amplification Burma padauk samplepsbK-psbIGene order fragment is as standard gene, and gene order is shown in sequence table (593 bases).
(2) usepsbK-psbIGene order identifies the method for Burma padauk, is divided into four steps:
The first step, samples and extracts DNA；
Second step, utilizes PCR reaction amplificationpsbK-psbIGene order；
3rd step, checks order to amplified production；
4th step, the sequence obtained is carried out sequence analysis with the psbK-psbI gene order in claim 1, when homology base ratio is less than 95%, get rid of the possibility that these species are Burma padauk, when homology base ratio more than or equal to 95% time, this sample be the probability of Burma padauk be × 100%.
Detailed description of the invention
In experiment, with the Burma padauk sample of 4 separate sources, extract sample DNA by TIANGENE company DNA extraction kit.The forward primer used: TTAGCCTTTGTTTGGCAAG, reverse primer: AGAGTTTGAGAGTAAGCAT, PCR amplification condition is 94 DEG C of denaturations 4min, following 94 DEG C of degeneration 45s, 50 DEG C of annealing 45 s, 72 DEG C extend 1.5min, carrying out altogether 35 circulations, last 72 DEG C extend 10min.Obtaining product comparison after order-checking, the homology between sample is held at more than 99%, it is thus achieved that sequence and other species are by online comparison, and its homology is all below 83%.
Sequence table:
AATTCCCCGAGCCTTACACTTAGATTTATTGGATTTGTTGCTAAAATATCGGTATTAAACCCGAAACTTCCGGCGGATGGCCAGTAACCCAAAGAAAGGAAAGAATCGGTTATATTTTTCATATGATCTCCTCTTATGGATAGACTAATTATCTTTCTACGATTCCGAATTTATTAGTATATATTACATATATATATATTATTGGTCGATTAATTGGATTGGAAGATTCCCCATAAGAATCATACTTATTAGAATTAGATACTAGTTCTTATGTCAATTGCAAAATCTCTAGTTGTTTTAAACGCTTTCCAAAAATTTTCTGAATGAAAAAAAGGGGGGGGGATTGGACTAAAAAGGGAAGGAATAAGGCAAGCAGTATGTTAATTTCTCACCTTAAATCAGTCCTTCCCCTGCCTTCCTGTACGAACGAATAAAAAATTGTAGGAGTGAAATCAAAATAATATAATTCGAAAAAGCAAGAGCAGCAAATCAAGTACACGGAAAAAGATATATGTATTTGTATTTTTCTATTTTTTTAGGGTTAAACAAAGGGATTCGCAAATAAAAGTGCTAATGCTACAACCAGCCCATAA。
Sequence table:
AATTCCCCGAGCCTTACACTTAGATTTATTGGATTTGTTGCTAAAATATCGGTATTAAACCCGAAACTTCCGGCGGATGGCCAGTAACCCAAAGAAAGGAAAGAATCGGTTATATTTTTCATATGATCTCCTCTTATGGATAGACTAATTATCTTTCTACGATTCCGAATTTATTAGTATATATTACATATATATATATTATTGGTCGATTAATTGGATTGGAAGATTCCCCATAAGAATCATACTTATTAGAATTAGATACTAGTTCTTATGTCAATTGCAAAATCTCTAGTTGTTTTAAACGCTTTCCAAAAATTTTCTGAATGAAAAAAAGGGGGGGGGATTGGACTAAAAAGGGAAGGAATAAGGCAAGCAGTATGTTAATTTCTCACCTTAAATCAGTCCTTCCCCTGCCTTCCTGTACGAACGAATAAAAAATTGTAGGAGTGAAATCAAAATAATATAATTCGAAAAAGCAAGAGCAGCAAATCAAGTACACGGAAAAAGATATATGTATTTGTATTTTTCTATTTTTTTAGGGTTAAACAAAGGGATTCGCAAATAAAAGTGCTAATGCTACAACCAGCCCATAA

Claims (2)

1. the standard gene for Burma padauk Molecular Identification, it is characterised in that its aminoacid sequence is the sequence in sequence table.
2. the method identifying Burma padauk 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, the sequence obtained is carried out sequence analysis with the psbK-psbI gene order in claim 1, when homology base ratio is less than 95%, get rid of the possibility that these species are Burma padauk, when homology base ratio more than or equal to 95% time, this sample be the probability of Burma padauk be × 100%.
CN201610326117.8A 2016-05-17 2016-05-17 Standard gene for molecular identification of pterocarpus macarocarpus kurz and molecular identification method Pending CN105925695A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110241243A (en) * 2019-06-14 2019-09-17 浙江省检验检疫科学技术研究院 The method for identifying molecules and its primer and probe of Ovshinsky yellow wingceltis

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CN103820569A (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 dalbergioides Benth.
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.
CN103834742A (en) * 2014-03-20 2014-06-04 中国科学院上海有机化学研究所 Special probe, primers, gene chip and method for detecting pterocarpus erinaceus poir.

Patent Citations (3)

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CN103820569A (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 dalbergioides Benth.
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.
CN103834742A (en) * 2014-03-20 2014-06-04 中国科学院上海有机化学研究所 Special probe, primers, gene chip and method for detecting pterocarpus erinaceus poir.

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Title
BENTE B. KLITGÅRD ET AL.: "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 (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110241243A (en) * 2019-06-14 2019-09-17 浙江省检验检疫科学技术研究院 The method for identifying molecules and its primer and probe of Ovshinsky yellow wingceltis
CN110241243B (en) * 2019-06-14 2024-02-27 浙江省检验检疫科学技术研究院 Molecular identification method of Pterocarpus of Orthosiphon and primer and probe thereof

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