CN104911256A - Chinese angelica SCAR molecular marker, its identification method and specific primer pair - Google Patents
Chinese angelica SCAR molecular marker, its identification method and specific primer pair Download PDFInfo
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Abstract
The invention relates to a Chinese angelica SCAR molecular marker, its nucleotide sequence is shown in a SEQ ID No.1. The invention also provides a PCR amplification specific primer pair used for detecting the Chinese angelica SCAR molecular marker, The nucleotide sequence of the primer pair is characterized in that an upstream primer is 5'-ACCCATTTGTAAACATAGCC-3' and a downstream primer is 5'- CAGATTCTCCTTCCACCC-3'. By using the molecular marker, certified products Chinese angelica medicinal material can be distinguished with medicinal material with similar form characteristic of fake stamps and substitutes, Chinese angelica SCAR molecular marker identification is carried out by using the primer and the method, PCR amplification specificity is good, in practical usage, screening of a lot of primers when the Chinese angelica is distinguished is not required, the identification method is rapid and simple, and the cost is low.
Description
Technical field
The present invention relates to biological technical field, concrete, relate to a kind of Radix Angelicae Sinensis SCAR molecule marker and authentication method thereof and Auele Specific Primer pair.
Background technology
When being classified as the dry root of umbelliferae angelica Angelica sinensis (Oliv.) Diels.Have and enrich blood, invigorate blood circulation, menstruction regulating and pain relieving, moisturize the effect of laxation, before more than 2,000 years, the ancient Chinese people have used its disease therapy, so far, Radix Angelicae Sinensis still has been widely used clinical, have saying of " ten side's rules for doing division with a one-digit divisor on the abacus ", therefore its market demand is very large, market is faced with the high-quality Radix Angelicae Sinensis situation that supply falls short of demand.Radix Angelicae Sinensis due to its use background and source complicated, domestic " homonym " used with the masterpiece Radix Angelicae Sinensis certified products of " Radix Angelicae Sinensis ", " Rhizome and root of Udo ", " wild Radix Angelicae Sinensis " is more, substitutes, phenomenon of passing a fake product off as a genuine one is further frequent.Wherein, mixed with adulterant with the substitute of the similar samphire of certified products Radix Angelicae Sinensis morphological specificity as the Chang Zuowei Radix Angelicae Sinensis such as Aba Radix Angelicae Sinensis, Angelica acutiloba, Root of Garden Lovage of the species of: heracleum hemsleyanum michaux, Bupleurum species, angelica kind, use among the people is more chaotic.Chemical composition and the certified products Radix Angelicae Sinensis of these plants differ greatly, and should be cautious use of.
The existing discriminating to Radix Angelicae Sinensis mainly adopts the traditional means such as microscopical identification and physics and chemistry discriminating.And current commercially available medicinal material mostly is processed goods, the surface texture of Radix Angelicae Sinensis and its mixed adulterant, even chemical composition is all extremely similar for microscopic features, to accurately differentiate that difficulty is very big in practice, and it is large to there is subjectivity, poor universality, and to defects such as viewer's skill requirement are high, be unfavorable for stdn, the foundation of normalization method and universal.DNA molecular Genetic Markers has fast, trace, accurately and the features such as high specificity, for the discriminating problem solving medicinal material provides objective and effective means.
Along with the development of molecular marking technique, the current minority bibliographical information Molecular Identification to Radix Angelicae Sinensis: Li Ming etc. (the peroxidase isozyme electrophoresis of Radix Angelicae Sinensis and mixed adulterant thereof differentiates [J]. Chinese medicinal materials, 2000,23 (4): 200.) utilize peroxidase isozyme to Radix Angelicae Sinensis and the qualification of mixed adulterant thereof, but this method is also only limitted to gather fresh plant carries out, and cannot differentiate pharmaceutical decocting piece.Feng etc. and this research group (FENG T, LIU S, HE XJ.Molecular authentication of the traditional Chinese medicinal plant Angelica sinensis based on internal transcribed spacer of nrDNA.Electronic Journal of Biotechnology, 2010,13 (1): 1-13.Zhang Chun, Wang Xiaoli, the big elder generation of tax, Zhu Ye, Zhuan Yuanchun. the rDNA ITS sequence of Chinese traditional medicine angelica and mixed adulterant thereof is analyzed and is differentiated, Sichuan Agricultural University's journal, 2011,29 (2): 218-224.) utilize ITS sequence to differentiate Radix Angelicae Sinensis and mixed adulterant thereof, but need to check order to sequence and cluster analysis can be distinguished.Discrimination process is comparatively loaded down with trivial details, and order-checking somewhat expensive, need the time longlyer to too increase testing cost.This research group utilizes ISSR technology to carry out Molecular Identification (Zhang Chun, Zhu Ye, the He Ying of Radix Angelicae Sinensis, Zhuan Yuanchun, the Radix Angelicae Sinensis Molecular Identification of series _ ISSR_ analysis is simply repeated based on inside. Chinese Pharmaceutical Journal, 2014,49 (10): 812-816).The method needs to increase to many primers, and carries out data analysis to electrophoretic band and just can draw reliable results.
To sum up, above-mentioned traditional discrimination method to Radix Angelicae Sinensis, accurately will differentiate that difficulty is very big, and it is large to there is subjectivity, poor universality in practice, and to defects such as viewer's skill requirement are high.And molecular identificalion means more accurately and reliably, but all need order-checking or a large amount of PCR and electrophoresis and later data process, be also unfavorable for that scale standardization is set up.
Therefore be necessary to develop a kind of result accurately and the simple and rapid molecular assay method of qualification process.
Summary of the invention
The object of the invention is to overcome the above-mentioned defect existed in prior art, provide a kind of result accurately sensitive, the simple and rapid Radix Angelicae Sinensis authentication method of qualification process.
For reaching above object, the present inventor, in a large amount of ISSR primer screening processes, finds out the specific amplification band of certified products Radix Angelicae Sinensis and is converted into SCAR mark, provides and differentiates comparatively fast to Radix Angelicae Sinensis, easy, cheap method.
The invention provides a kind of Radix Angelicae Sinensis SCAR molecule marker, its nucleotide sequence is as shown in SEQ ID No:1.
Present invention also offers a kind of pcr amplification Auele Specific Primer pair for detecting Radix Angelicae Sinensis SCAR molecule marker of the present invention, the nucleotides sequence of described primer pair is classified as:
Upstream primer: 5 '-ACCCATTTGTAAACATAGCC-3 ' (shown in SEQ ID No:2 sequence)
Downstream primer: 5 '-CAGATTCTCCTTCCACCC-3 ' (shown in SEQ ID No:3 sequence).
In the design of primers stage, contriver considers in actual application, and amplification is easy to carry out and specificity is better, and amplification section is more preferably greater than 500bp.In order to reduce primer synthesis cost, primer length is limited in about 20bp, and end can not contain more than 3 continuous bases, particularly AAA and TTT.Can not form dimer between primer, self can not form hairpin structure, and annealing temperature can not lower than 50 degree.Inventor has devised 10 pairs of primers according to above principle, and respectively the object fragment amplification effect of these 10 pairs of primers is verified, find to achieve best expanding effect compared with pcr amplification Auele Specific Primer provided by the present invention and other 9 pairs of primers.The wherein sequence of a pair of 10 pairs of such as involved primers is:
826L:5’-GTAAACATAGCCGCACAAA-3’，
826R:5’-CTGCCAAGCCTCACAAAC-3’。
But to having there are 2 amplified bands in this, specificity is good not, is therefore eliminated after primer amplification.
Another aspect of the present invention provides a kind of authentication method of Radix Angelicae Sinensis SCAR molecule marker, and described method comprises: with the STb gene of testing sample for template, utilizes primer pair provided by the present invention to carry out pcr amplification reaction, and carries out electrophoresis detection to amplified production.
Optionally, the system of described pcr amplification reaction is:
Optionally, the program of described pcr amplification reaction is:
95 DEG C of denaturation 4min; 94 DEG C of sex change 40s, 60 DEG C of annealing 40s, 72 DEG C extend 1min, totally 35 circulations; 8min is extended after 72 DEG C.
The present invention can obtain the best identification result to Radix Angelicae Sinensis SCAR molecule marker by above-mentioned primer, reaction system, response procedures, can Radix Angelicae Sinensis and other samples be separated more quickly and accurately.
Optionally, described testing sample is selected from one or more in Radix Angelicae Sinensis, Aba Radix Angelicae Sinensis, Angelica acutiloba, Root of Garden Lovage, RADIX PEUCEDANI, the root of Dahurain angelica, Radix Bupleuri, white bright levisticum, levisticum, Jehol Ligusticum Rhizome, Ligusticum wallichii, Radix Chuanminshen, notopterygium root, omei angelica root, sweet fennel, windproof, Stem and leaf of Japanese Cryptotaenia Cryptotaenia japonica Hassk. and Daucus carota L..
Present invention also offers a kind of test kit for the identification of Radix Angelicae Sinensis, comprise Auele Specific Primer pair provided by the present invention.
Utilize molecule marker provided by the present invention, accurately floral region similar to morphological specificity for certified products Radix Angelicae Sinensis can be separated, primer provided by the present invention and method is utilized to carry out Radix Angelicae Sinensis SCAR molecular markers for identification, the specificity of pcr amplification is good, in actual applications, without the need to screening a large amount of primers when Radix Angelicae Sinensis is differentiated, also without the need to genetic similarity and the structure genetic cluster collection of illustrative plates of the data estimation sample room according to acquisition, have fast, the superiority of easy and low cost.
Accompanying drawing explanation
Fig. 1 is for using primer UBC826 to Radix Angelicae Sinensis and mixed adulterant 29 samples amplification electrophorograms thereof
M:DL2000DNA marker, numeral number is with table 1, and arrow indication is Radix Angelicae Sinensis specific amplified band.
Fig. 2 Auele Specific Primer provided by the present invention is to the amplification electrophorogram to Radix Angelicae Sinensis and mixed adulterant 29 samples thereof
M:DL2000DNA marker, numeral number is with table 1.
Embodiment
Below will the present invention is described in detail by embodiment.
Embodiment 1
The extraction of 1.1 DNA and detection
Get 10mg and in liquid nitrogen, grind to form fine powder through the quick-drying detected materials of silica gel, use DNA extraction kit (sky, Beijing root DP305) to extract STb gene.Cross 1% gel electrophoresis, the integrity of full automatic gel imaging system analyzing DNA and purity.Its concentration, OD is measured with micro-ultraviolet-visible spectrophotometer (NanoDrop ND1000)
260/ OD
280value.
1.2 pcr amplifications and electrophoretic analysis detect
Factor on primer sharpness and polymorphism may be affected: the content of DNA profiling, MgCl
2concentration, dNTP and Taq enzyme content, annealing temperature etc., utilize primer UBC826, UBC825 and UBC 857 is the representative (object of this step: because ISSR primer has 100, contriver have selected Radix Angelicae Sinensis before this, Aba Radix Angelicae Sinensis and Root of Garden Lovage three parts of materials screen these 100 pairs of primers, selection amplified band is many, clear, bright primer carries out subsequent analysis, refer to document (Zhang Chun, Zhu Ye, He Ying, Zhuan Yuanchun, the Radix Angelicae Sinensis Molecular Identification of series _ ISSR_ analysis is simply repeated based on inside. Chinese Pharmaceutical Journal, 2014, 49 (10): 812-816).Primer UBC826, UBC825 and UBC 857 is band number comparatively horn of plenties after amplification, and more clear bright primer, therefore select these primers as the optimization of PCR condition, as adjustment template, dNTP, MgCl
2and primer concentration and Taq enzyme content etc., more reliable and stable to reach amplification, and band number is many and clear.The optimum primer screened and optimum PCR condition is utilized to carry out amplification and the analysis of 29 parts of materials afterwards.), PCR reaction system is optimized.Finally determine that the reaction system after PCR optimization is 20 μ L, comprising 10 × reaction buffer 2.0 μ L, dNTP 200 μm of olL
-1, MgCl
21.5mmolL
-1, primer 1.0 μm of olL
-1, 30ng DNA masterplate, Ex Taq enzyme 1.25U (TaKaRa).PCR response procedures is: 95 DEG C of denaturation 4min; 94 DEG C of sex change 40s, 48-55 DEG C of annealing 40s, 72 DEG C extend 1min, totally 35 circulations; 8min is extended, 4 DEG C of preservations after 72 DEG C.PCR primer is electrophoresis (1 × TBE) on the sepharose of 1.5%, and band utilizes gel imaging system to observe, take pictures after being separated.
The acquisition of 1.3 ISSR primer screenings and Radix Angelicae Sinensis specific band
Article 100, ISSR primer adopts Canadian Columbia University (Biotechnology Laboratory, University of British Columbia, Vancouver, Canada, http://www.ubc.ca/) sequence that provides, (Zhao Qian, Du Hong, the village red .ISSR molecule marker in east and the application in plant research thereof. Molecular Plant Breeding, 2007,5 (6): 123-129.) to amplify clearly according to optimum PCR screening, repeatably and the amplification of primer for all samples of 3-15 pleomorphism site can be produced.(UBC 826 is Canadian Columbia University (Biotechnology Laboratory to primer UBC 826, University of British Columbia, Vancouver, Canada, http://www.ubc.ca/) in 100 ISSR primer sequences announcing one.) obtain the peculiar band (see Fig. 1 arrow Suo Shi) of certified products Radix Angelicae Sinensis after amplification 29 parts of materials.This band is cut under ultraviolet lamp, after fragment reclaims, utilizes pGM-T carrier to connect.Product conversion escherichia coli DH5a after connection.Selecting 3-5 positive bacterium colony send biotech firm to check order.Order-checking obtains consistent results, and its sequence length is 899bp.Utilize upstream primer: 5 '-ACCCATTTGTAAACATAGCC-3 ' (shown in SEQ ID No:2 sequence), downstream primer: 5 '-CAGATTCTCCTTCCACCC-3 ' (shown in SEQ ID No:3 sequence), increase this sequence, and amplification length is that its sequence of 749bp is in table 1.
Embodiment 2
29 parts of materials that the primer pair utilizing specific fragment provided by the present invention to design is collected carry out the specificity that pcr amplification detects this fragment.Comprising 6 parts from the certified products Radix Angelicae Sinensis of Different sources and the mixed adulterant of 23 parts of Radix Angelicae Sinensis and relative genus plant (see table 1).Pcr amplification condition is: cumulative volume 20 μ L, comprising 10 × reaction buffer 2.0 μ L, dNTP 200 μm of olL
-1, MgCl
21.5mmolL
-1, primer 1.0 μm of olL
-1, 30ng DNA masterplate, Ex Taq enzyme 1.25U (TaKaRa).PCR response procedures is: 95 DEG C of denaturation 4min; 94 DEG C of sex change 40s, 60 DEG C of annealing 40s, 72 DEG C extend 1min, totally 40 circulations; 8min is extended, 4 DEG C of preservations after 72 DEG C.Detected result display (see Fig. 2), only have certified products Radix Angelicae Sinensis can amplify the band consistent with object clip size, other species all can not amplify band.Show after this primer amplification, its dimer is less, without non-specific amplification band.Therefore this mark is easy to amplification, and its specificity is better, can as the special SCAR mark of Radix Angelicae Sinensis real and fake discrimination.In actual applications, without the need to screening a large amount of primers when Radix Angelicae Sinensis is differentiated, also without the need to according to the genetic similarity of the data estimation sample room obtained with build genetic cluster collection of illustrative plates, have fast, the superiority of easy and low cost.
Table 1
The special SCAR mark of Radix Angelicae Sinensis provided by the present invention, only need to carry out DNA extraction to sample, react with One_step PCR, namely Radix Angelicae Sinensis is differentiated fast by electrophoresis result, without the need to carrying out band statistics and data analysis, and 29 parts of materials of kindred plant are analyzed, still there is good specificity, therefore can as the method for standardization qualification Radix Angelicae Sinensis.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (7)
1. a Radix Angelicae Sinensis SCAR molecule marker, is characterized in that, its nucleotide sequence is as shown in SEQ ID No:1.
2. require the pcr amplification Auele Specific Primer pair of the Radix Angelicae Sinensis SCAR molecule marker described in 1 for test right, it is characterized in that, the nucleotides sequence of described primer pair is classified as:
Upstream primer: 5 '-ACCCATTTGTAAACATAGCC-3 '
Downstream primer: 5 '-CAGATTCTCCTTCCACCC-3 '.
3. the authentication method of Radix Angelicae Sinensis SCAR molecule marker according to claim 1, it is characterized in that, described method comprises: with the STb gene of testing sample for template, utilizes the primer pair described in claim 2 to carry out pcr amplification reaction, and carries out electrophoresis detection to amplified production.
4. authentication method according to claim 3, is characterized in that, the system of described pcr amplification reaction is:
5. the authentication method according to claim 3 or 4, is characterized in that, the program of described pcr amplification reaction is:
95 DEG C of denaturation 4min; 94 DEG C of sex change 40s, 60 DEG C of annealing 40s, 72 DEG C extend 1min, totally 35 circulations; 8min is extended after 72 DEG C.
6. authentication method according to claim 5, it is characterized in that, described testing sample be selected from Radix Angelicae Sinensis, Aba Radix Angelicae Sinensis, Angelica acutiloba, Root of Garden Lovage, RADIX PEUCEDANI, the root of Dahurain angelica, Radix Bupleuri, white bright levisticum, levisticum, Jehol Ligusticum Rhizome, Ligusticum wallichii, Radix Chuanminshen, notopterygium root, omei angelica root, sweet fennel, windproof, Stem and leaf of Japanese Cryptotaenia Cryptotaenia japonica Hassk. and Daucus carota L. one or more.
7. for the identification of a test kit for Radix Angelicae Sinensis, it is characterized in that, comprise Auele Specific Primer pair according to claim 2.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104946636A (en) * | 2015-06-30 | 2015-09-30 | 四川医科大学 | Levisticum officinale detection kit and detection method |
| CN104946635A (en) * | 2015-06-30 | 2015-09-30 | 四川医科大学 | Detection kit and detection method for ligusticum acutilobum |
| CN106434910A (en) * | 2016-09-20 | 2017-02-22 | 广东东阳光药业有限公司 | Primer and amplification system for identifying cordyceps sinensis and identification method |
| CN107130038A (en) * | 2017-06-07 | 2017-09-05 | 苏州市李良济健康产业有限公司 | A kind of primer pair and its application for Identification chinese herbs medicine Radix Angelicae Sinensis |
| CN109666758A (en) * | 2019-02-14 | 2019-04-23 | 四川省农业科学院经济作物育种栽培研究所 | Genuine rhizome of chuanxiong identification primer and identification method based on InDel label |
| CN110241247A (en) * | 2019-07-12 | 2019-09-17 | 华润三九医药股份有限公司 | A kind of molecular labeling, primer pair and discrimination method identified for tripterygium wilfordii |
| CN112553364A (en) * | 2020-12-25 | 2021-03-26 | 湖北省农业科学院中药材研究所 | ISSR-PCR molecular marker of Angelica crenulata and application thereof |
| CN114317799A (en) * | 2021-12-21 | 2022-04-12 | 中国中医科学院中药研究所 | Specific primer pair for identifying angelica and common angelica mixed counterfeit and application thereof |
| CN114457183A (en) * | 2022-02-21 | 2022-05-10 | 广州白云山光华制药股份有限公司 | SCAR molecular marker, specific primer pair and method for identifying Xikangchui |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104004818A (en) * | 2013-02-26 | 2014-08-27 | 中国医学科学院药用植物研究所 | Method for identifying Chinese angelica and its adulterants by utilizing ITS sequence |
-
2015
- 2015-03-10 CN CN201510104132.3A patent/CN104911256B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104004818A (en) * | 2013-02-26 | 2014-08-27 | 中国医学科学院药用植物研究所 | Method for identifying Chinese angelica and its adulterants by utilizing ITS sequence |
Non-Patent Citations (3)
| Title |
|---|
| ZHIQIANG MEI等: "Efficiency of improved RAPD and ISSR markers in assessing genetic diversity and relationships in Angelica sinensis (Oliv.) Diels varieties of China", 《ELECTRONIC JOURNAL OF BIOTECHNOLOGY》 * |
| 张春等: "基于内部简单重复系列(ISSR)分析的当归分子鉴定", 《中国药学杂志》 * |
| 曹亮等: "SRAP和SCAR分子标记应用于中药", 《科技导报》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104946636A (en) * | 2015-06-30 | 2015-09-30 | 四川医科大学 | Levisticum officinale detection kit and detection method |
| CN104946635A (en) * | 2015-06-30 | 2015-09-30 | 四川医科大学 | Detection kit and detection method for ligusticum acutilobum |
| CN104946636B (en) * | 2015-06-30 | 2018-01-02 | 四川医科大学 | The detection kit and detection method of lovage |
| CN104946635B (en) * | 2015-06-30 | 2018-05-22 | 四川医科大学 | The detection kit and detection method of ligusticum acutilobum |
| CN106434910A (en) * | 2016-09-20 | 2017-02-22 | 广东东阳光药业有限公司 | Primer and amplification system for identifying cordyceps sinensis and identification method |
| CN107130038A (en) * | 2017-06-07 | 2017-09-05 | 苏州市李良济健康产业有限公司 | A kind of primer pair and its application for Identification chinese herbs medicine Radix Angelicae Sinensis |
| CN109666758A (en) * | 2019-02-14 | 2019-04-23 | 四川省农业科学院经济作物育种栽培研究所 | Genuine rhizome of chuanxiong identification primer and identification method based on InDel label |
| CN110241247A (en) * | 2019-07-12 | 2019-09-17 | 华润三九医药股份有限公司 | A kind of molecular labeling, primer pair and discrimination method identified for tripterygium wilfordii |
| CN110241247B (en) * | 2019-07-12 | 2023-06-20 | 华润三九医药股份有限公司 | Molecular marker and primer pair for identifying tripterygium wilfordii medicinal materials and identification method |
| CN112553364A (en) * | 2020-12-25 | 2021-03-26 | 湖北省农业科学院中药材研究所 | ISSR-PCR molecular marker of Angelica crenulata and application thereof |
| CN114317799A (en) * | 2021-12-21 | 2022-04-12 | 中国中医科学院中药研究所 | Specific primer pair for identifying angelica and common angelica mixed counterfeit and application thereof |
| CN114317799B (en) * | 2021-12-21 | 2023-10-13 | 中国中医科学院中药研究所 | Specific primer pair for identifying angelica and common angelica mixed and counterfeited products and application thereof |
| CN114457183A (en) * | 2022-02-21 | 2022-05-10 | 广州白云山光华制药股份有限公司 | SCAR molecular marker, specific primer pair and method for identifying Xikangchui |
| CN114457183B (en) * | 2022-02-21 | 2024-04-16 | 广州白云山光华制药股份有限公司 | SCAR molecular marker for identifying Western Kang Chaihu, specific primer pair and method |
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