CN110714094B - SCAR molecular marker for specifically identifying tremella strain and identification method and application thereof - Google Patents
SCAR molecular marker for specifically identifying tremella strain and identification method and application thereof Download PDFInfo
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Abstract
The invention provides an SCAR molecular marker for specifically identifying tremella strains, an identification method and application thereof, wherein the nucleotide sequence of the SCAR molecular marker is shown as SEQ ID NO: 1, or a sequence similar to that shown in SEQ ID NO: 1, or is consistent with the middle sequence within 30bp of the front end and the back end in the sequence shown in SEQ ID NO: the similarity of the middle sequences in 30bp at the front end and the rear end in the sequence shown in 1 reaches more than 99 percent. The invention also provides a primer designed based on the SCAR molecular marker, different tremella varieties can be identified by using the SCAR molecular marker and the primer, the tremella TWW01-AX strain can be identified quickly and effectively, and the primers have the advantages of simple operation, short time consumption, strong specificity and the like.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to an SCAR molecular marker for specifically identifying a tremella strain, and an identification method and application thereof.
Background
Tremella fuciformis (Tremella fuciformis) belongs to Tremellales (Tremellales), Tremellaceae (Tremellaceae) and Tremella (Tremella) and is a famous food and medicinal fungus in China. The tremella contains various bioactive substances, can improve the immunity of organisms and relieve the damage of nervous systems, has the effects of resisting tumors, radiation, oxidation and aging, reducing blood sugar and blood fat and the like, and has wide development prospect.
Tremella fuciformis is used as an edible fungus variety with great characteristic advantages in China, and the yield of the tremella fuciformis accounts for more than 90% of the world. At present, the main cultivated species of tremella in China is single, and the main cultivated species has degeneration with different degrees along with the increase of the number of passages, which brings certain risks to the seed production work and urgently needs the supplement of high-quality new species. Compared with other edible fungi, the folk tremella seed production is disordered, the dispute on the strain source is large, and the problems of homonymy and heteronymy, homonymy and heteronymy and the like are also outstanding. In addition, with the further expansion of the cultivation scale, the demand for high-quality tremella cultivation strains which can adapt to different altitudes and different climatic conditions is increasing, so that a simple, convenient, quick and accurate-result strain identification technology needs to be established, so as to ensure the accurate use of the strains, protect the benefits of breeders and producers and avoid the waste of scientific research resources.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide an SCAR molecular marker for specifically identifying a tremella strain TWW01-AX, wherein the nucleotide sequence of the SCAR molecular marker is shown as SEQ ID NO: 1, or a sequence similar to that shown in SEQ ID NO: 1, or is consistent with the middle sequence within 30bp of the front end and the back end in the sequence shown in SEQ ID NO: the similarity of the middle sequences in 30bp at the front end and the rear end in the sequence shown in 1 reaches more than 99 percent.
In the sequence synthesis process, the synthetic sequences are different due to the change of the synthetic system or program, but according to the invention, the sequence mainly shows that partial base substitution, insertion or deletion can occur in the sequence of about 30bp at the front end and the rear end, but only the intermediate sequence within about 30bp at the front end and the rear end, namely the sequence based on SEQ ID NO: 1, the sequence of about 30bp at the front end or about 30bp at the back end in the sequence can be changed, but the sequences of the middle fragments at the two ends are not changed, or the similarity of the middle sequences reaches more than 99 percent, which is in the protection scope of the invention.
The tremella strain TWW01-AX is preserved in China general microbiological culture Collection center in 2019, 11 months and 18 days, and the preservation number is CGMCC NO. 18809.
Another objective of the invention is to provide a primer designed based on the SCAR molecular marker, wherein the primer sequence is preferably as follows:
2047-S-1F：5'-GGCGGAGGATACAGATTG-3'；
2047-S-1R：5'-CCAGTAGGTAGCGGAAGA-3'。
the third purpose of the invention is to use the SCAR molecular marker or primer for identifying the tremella strain TWW 01-AX.
The method for identifying the tremella strain TWW01-AX by adopting the SCAR molecular marker or the primer comprises the following steps:
extracting sample DNA, carrying out SCAR-PCR amplification on the sample DNA, and if a band of 308bp is amplified, indicating that the sample strain is a tremella strain TWW 01-AX.
The SCAR-PCR reaction system is as follows: 1 uL of each of the upstream and downstream primers, 1 uL of DNA template, 12.5 uL of 2 × EasyTaq PCR Supermix, ddH2O9.5 mu L; the SCAR-PCR reaction program is as follows: pre-denaturation at 94 deg.C for 5min, denaturation at 94 deg.C for 1min, annealing at 55 deg.C for 1min, extension at 72 deg.C for 1min, 30 cycles, and extension at 72 deg.C for 10 min.
The SCAR molecular marker for specifically identifying the tremella strain, the identification method and the application thereof provided by the invention have the following beneficial effects:
the molecular marker and the primers of the SCAR can be used for quickly and effectively identifying the tremella TWW01-AX strain, and have the advantages of simple operation, short consumed time, strong specificity and the like.
Drawings
FIG. 1 shows the amplification results of random polymorphic amplification of different Tremella strains with primer S2047.
FIG. 2 is the result of PCR amplification of different Tremella strains using specific SCAR primers.
Detailed Description
The technical scheme of the invention is a conventional scheme in the field if not specifically stated; the reagents or materials, if not specifically mentioned, are commercially available.
The present invention will be further described with reference to the following examples.
Example 1 obtaining of SCAR molecular marker primers for specific identification of Tremella fuciformis strains
(1) Culture of tremella strain saccharomycete spore and extraction of DNA
The name and source of the tremella strain are shown in Table 1, and the yeast-like spores of different tremella strains stored at low temperature are activated in a rotating tube, activated and cultured at 25 ℃ for one week, then inoculated into a PDA liquid culture medium, and cultured in a constant-temperature shaking incubator at 25 ℃ and 150r/min for 7 days. After the culture is finished, the thalli sediment is centrifugally collected under the aseptic condition, and the genome DNA of each tremella strain is extracted by adopting an Ezup column type fungus genome DNA extraction kit of biological engineering (Shanghai) GmbH.
TABLE 1 Tremella fuciformis strain name and source
(2) Obtaining of Tremella strain TWW01-AX specific DNA fragment
Screening Random Amplification Polymorphic DNA (RAPD) primers by adopting a PCR technology, and finally carrying out random polymorphic amplification on each tremella strain by using a primer S2047(5'-TGTGCCTGAC-3', SEQ ID NO: 4), wherein the RAPD reaction system is as follows: primer 1. mu.L, DNA template 1. mu.L, 2 × EasyTaq PCR Supermix (Beijing Quanjin Biotechnology Co., Ltd.) 12.5. mu.L, ddH2O10.5 μ L, RAPD amplification reaction program: pre-denaturation at 94 deg.C for 5min, denaturation at 94 deg.C for 1min, annealing at 35 deg.C for 1.5min, extension at 72 deg.C for 1.5min, 30 cycles, extension at 72 deg.C for 10min, and storage at 4 deg.C. The amplification products were subjected to gel electrophoresis using 1% agarose and 120V voltage, and photographed by observation using a gel image analyzer, as shown in FIG. 1 (lanes 1 to 11, T23-CAS, T26-CAS, T30-CAS, T33-CAS, T56-CAS, T74-CAS, T8225-CAS, TWW01-AX, TY01-SC, TR01, and TR21, respectively). The RAPD amplification result shows that obvious polymorphism exists between test strains, and the primer S2047 can amplify obvious specific fragments in TWW01-AX strain (shown by arrows in the figure).
(3) Recovery, cloning, sequencing of specific fragments
The specific fragment obtained in step (2) was recovered and purified using Gel Extraction Kit D2500 Gel recovery Kit from OMEGA. The recovered target DNA was ligated to pMD18-T Vector (Baozi's physician's technology (Beijing) Co., Ltd.), transformed into Trans1-T1 competent cells (Beijing Quanjin Biotechnology Co., Ltd.), plated on LB solid medium containing IPTG, X-gal, AMP, and cultured overnight at 37 ℃. And (3) selecting a white single colony, inoculating the white single colony into an LB liquid culture medium, culturing for 12-16 hours, extracting a plasmid, and performing double enzyme digestion verification by using EcoR 1 and Hind III. The successfully transformed transformants were sent to the Biotechnology engineering (Shanghai) GmbH for sequencing.
(4) Design of specific SCAR-PCR amplification primer pair
Designing a pair of specific primers for SCAR reaction by using Primer Premier 6 software according to the sequence of the specific fragment obtained in the step (3), wherein the sequences of the primers are as follows:
2047-S-1F：5'-GGCGGAGGATACAGATTG-3'(SEQ ID NO：2)，
2047-S-1R: 5'-CCAGTAGGTAGCGGAAGA-3' (SEQ ID NO: 3), synthesized by Biotechnology engineering (Shanghai) GmbH.
(5) SCAR-PCR amplification
And (5) carrying out SCAR-PCR amplification verification on the genome DNA of each tremella strain by using the specific amplification primer pair obtained in the step (4). The SCAR-PCR reaction system is as follows: 1. mu.L of each of the upstream and downstream primers, 1. mu.L of DNA template, 12.5. mu.L of 2 × EasyTaq PCR Supermix (Beijing Quanyujin Biotechnology Co., Ltd.), and ddH2O9.5. mu.L. The SCAR-PCR reaction program is as follows: pre-denaturation at 94 deg.C for 5min, denaturation at 94 deg.C for 1min, annealing at 55 deg.C for 1min, extension at 72 deg.C for 1min, 30 cycles, extension at 72 deg.C for 10min, and storage at 4 deg.C.
(6) Electrophoretic detection
And (5) detecting the SCAR-PCR amplification product in the step (5) by using 1% agarose gel electrophoresis at the voltage of 120V, and photographing and observing on a gel imaging analyzer after the electrophoresis is finished. As shown in FIG. 2, a 308bp specific band was detected in the TWW01-AX lane of Tremella strain, while no specific fragment was found in the same position in other Tremella species.
(7) Sequencing the specific DNA fragment of the tremella strain TWW01-AX amplified in the step (6), wherein the sequence information of the amplified specific fragment is as follows: 5'-TCCAGTAGGTTGGGAAGACGGCGTCAGTGCTTGTCGTTAGACTGATCTTTCAACATGGAGACAGCTTACAAAGGCCCCGGGCCCGTCAAATCGATCTATCTGTCAGCTACCATCCCGAGTATCATCGCTGAGCGCTCACTGCTCCAAGCGATCCTACGCCTCCGTTTTCGACTAGCTCAACGATGGTGGCGATGGTGTCCAGCAGGATCGGGTGGAACGGTTTGGACTGGATGTCAGCGCTCGATCCCATTCGCAGCCAGCCATAGCGCACCACCATGGTATACTGCCCAATCTCATCCTCCGCCCAC-3' (SEQ ID NO: 1).
Sequence listing
<110> Fujian agriculture and forestry university
<120> SCAR molecular marker for specifically identifying tremella strain, and identification method and application thereof
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 308
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
tccagtaggt tgggaagacg gcgtcagtgc ttgtcgttag actgatcttt caacatggag 60
acagcttaca aaggccccgg gcccgtcaaa tcgatctatc tgtcagctac catcccgagt 120
atcatcgctg agcgctcact gctccaagcg atcctacgcc tccgttttcg actagctcaa 180
cgatggtggc gatggtgtcc agcaggatcg ggtggaacgg tttggactgg atgtcagcgc 240
tcgatcccat tcgcagccag ccatagcgca ccaccatggt atactgccca atctcatcct 300
ccgcccac 308
<210> 2
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
ggcggaggat acagattg 18
<210> 3
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
ccagtaggta gcggaaga 18
<210> 4
<211> 10
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Claims (6)
1. A SCAR molecular marker for specifically identifying a tremella strain TWW01-AX is characterized in that the nucleotide sequence of the SCAR molecular marker is shown as SEQ ID NO: 1 is shown.
2. A primer designed based on the SCAR molecular marker of claim 1, wherein the primer is:
2047-S-1F：5'-GGCGGAGGATACAGATTG-3'；
2047-S-1R：5'-CCAGTAGGTAGCGGAAGA-3'。
3. use of the SCAR molecular marker of claim 1 or the primer of claim 2 for identifying Tremella fuciformis strain TWW 01-AX.
4. The method for identifying the tremella strain TWW01-AX by using the SCAR molecular marker of claim 1 or the primer of claim 2, which is characterized by comprising the following steps:
extracting sample DNA, carrying out SCAR-PCR amplification on the sample DNA, and if a band of 308bp is amplified, indicating that the sample strain is a tremella strain TWW 01-AX.
5. The method for identifying the tremella strain TWW01-AX as claimed in claim 4, wherein the SCAR-PCR reaction system is: mu.L of each of the upstream and downstream primers, 1. mu.L of DNA template, 12.5. mu.L of Supermix for 2 × EasyTaq PCR, and 9.5. mu.L of ddH2O 9.5.
6. The method for identifying the tremella strain TWW01-AX as claimed in claim 4, wherein the SCAR-PCR reaction procedure is: pre-denaturation at 94 deg.C for 5min, denaturation at 94 deg.C for 1min, annealing at 55 deg.C for 1min, extension at 72 deg.C for 1min, 30 cycles, and extension at 72 deg.C for 10 min.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995002691A2 (en) * | 1993-07-13 | 1995-01-26 | Bromyc B.V. | Production and application of transgenic mushroom mycelium and fruitbodies |
| CN101086017A (en) * | 2007-04-29 | 2007-12-12 | 上海市农业科学院食用菌研究所 | Mushroom 507 bacteria molecular specific mark and its obtaining method and uses |
| CN101475991A (en) * | 2009-01-19 | 2009-07-08 | 黑龙江省科学院微生物研究所 | Method for identifying black fungus bacterial strain 185 and gene sequence for identifying black fungus bacterial strain 185 |
| CN101798601A (en) * | 2010-04-28 | 2010-08-11 | 黑龙江省科学院微生物研究所 | Molecular marker identification method of black fungus strain and specific molecular marker primers of black fungus strain HW 5 |
| CN109182581A (en) * | 2018-10-10 | 2019-01-11 | 大闽食品（漳州）有限公司 | A method of the Siraitia grosvenorii male and female plant Rapid identification based on specific molecular marker |
-
2019
- 2019-11-22 CN CN201911157587.6A patent/CN110714094B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995002691A2 (en) * | 1993-07-13 | 1995-01-26 | Bromyc B.V. | Production and application of transgenic mushroom mycelium and fruitbodies |
| CN101086017A (en) * | 2007-04-29 | 2007-12-12 | 上海市农业科学院食用菌研究所 | Mushroom 507 bacteria molecular specific mark and its obtaining method and uses |
| CN101475991A (en) * | 2009-01-19 | 2009-07-08 | 黑龙江省科学院微生物研究所 | Method for identifying black fungus bacterial strain 185 and gene sequence for identifying black fungus bacterial strain 185 |
| CN101798601A (en) * | 2010-04-28 | 2010-08-11 | 黑龙江省科学院微生物研究所 | Molecular marker identification method of black fungus strain and specific molecular marker primers of black fungus strain HW 5 |
| CN109182581A (en) * | 2018-10-10 | 2019-01-11 | 大闽食品（漳州）有限公司 | A method of the Siraitia grosvenorii male and female plant Rapid identification based on specific molecular marker |
Non-Patent Citations (4)
| Title |
|---|
| Applied modern biotechnology for cultivation ofGanodermaand development of their products;Xuan-Wei Zhou，等;《Appl Microbiol Biotechnol》;20111215;第93卷(第3期);第941-963页 * |
| SRAP、ISSR和RAPD分子标记技术在银耳菌株鉴别上的应用;曲绍轩;《食用菌学报》;20070915;第14卷(第3期);第1-5页 * |
| 银耳芽孢遗传多样性分析;孙淑静，等;《热带作物学报》;20090925;第30卷(第09期);第1308-1313页 * |
| 黑龙江省野生黑木耳菌种的ISSR指纹分析及SCAR标记;王立枫;《中国优秀硕士学位论文全文数据库（硕士）农业科技辑》;20110515(第2011年05期);第D048-48页 * |
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