CN108456742B - Molecular marker of oudemansiella lodana JZB2115055 and application thereof - Google Patents
Molecular marker of oudemansiella lodana JZB2115055 and application thereof Download PDFInfo
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Abstract
The invention discloses a molecular marker of oudemansiella lodana JZB2115055 and application thereof. The invention provides a specific primer of oudemansiella lodestone JZB2115055, which can amplify DNA including DNA molecule shown in SEQ ID No.2 from genome DNA of oudemansiella lodestone JZB 2115055. Experiments prove that the DNA molecule shown in SEQ ID No.3, the DNA molecule shown in SEQ ID No.2 or the DNA molecule shown in SEQ ID No.1 is a specific molecular marker of Oudemansiella (Oudemansiella canarii) JZB2115055, and can be used for rapidly identifying Oudemansiella (Oudemansiella canarii) JZB 2115055.
Description
Technical Field
The invention relates to a molecular marker of oudemansiella lodamiana JZB2115055 in the field of biotechnology and application thereof.
Background
Oudemansiella (Oudemansiella) belongs to the Basidiomycota (Basidiomycota) family, the Sarcophythoraceae family (Physalacriceae). The genus Oudemansia includes various species such as Oudemansia punctata (Oudemansiella radiata), Oudemansia fusca (Oudemansiella brunneomarginata), Pleurotus citrinopileatus (Oudemansiella venosollamata), Oudemansia mucosae (Oudemansiella mucida), and Oudemansia ochracea (Oudemansiella canarii). Wherein, the Oudemansiella fusceolata is also called Tropical Oudemansiella, and the fruiting body is rich in protein, carbohydrate, various vitamins and mineral elements. Chinese patent application publication No. CN105769938A and application No. 201610245029.5, published on 20/7/2016, disclose Oudemansiella canarii (Oudemansiella canarii) with strain No. JZB2115055, and the flavone extract and polysaccharide extract of the strain have effects of scavenging free radicals and resisting oxidation.
The conventional edible fungus strain identification mainly comprises morphological identification, somatic cell incompatibility identification (antagonism identification), ITS identification, fruiting comparison and the like. The appearance shape is easily influenced by a cultivation formula and a cultivation environment, so that the identification of the appearance shape is difficult; antagonistic lines between strains with relatively close relationships are not obvious, and are difficult to distinguish through ITS sequences; the fruiting contrast experiment takes long time, at least 2-3 months, which is not beneficial to the rapid identification and detection of strains. With the development of molecular biology technology, molecular marking methods are more and more widely used for identifying strains of edible fungi.
Disclosure of Invention
The technical problem to be solved by the invention is how to identify Oudemansiella canarii (Oudemansiella canarii) JZB 2115055.
In order to solve the technical problems, the invention provides a specific molecular marker of Oudemansiella canarii (Oudemansiella canarii) JZB 2115055.
The specific molecular marker of Oudemansiella canarii JZB2115055 provided by the invention is DNA shown as sequence 1, sequence 2 or sequence 3 in a sequence table.
In the specific molecular marker, the size of the DNA shown in the sequence 1 in the sequence table is 753 bp; the size of DNA shown in a sequence 2 in the sequence table is 537 bp; the DNA size shown in sequence 3 in the sequence table is 431 bp.
In order to solve the technical problems, the invention provides specific primers of Oudemansiella canarii (Oudemansiella canarii) JZB 2115055.
The specific primer of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 provided by the invention can be used for amplifying DNA (deoxyribonucleic acid) including a DNA molecule shown in SEQ ID No.2 from the genomic DNA of the Oudemansiella ochracea (Oudemansiella canarii) JZB 2115055.
In the specific primer, the specific primer can be composed of P826-1-2XF and P826-1-2 XR; the P826-1-2XF is a single-stranded DNA molecule shown in SEQ ID No. 4; the P826-1-2XR is a single-stranded DNA molecule shown in SEQ ID No. 5. The P826-1-2XF and the P826-1-2XR can amplify DNA molecules shown as SEQ ID No.2 and SEQ ID No.3 from the genomic DNA of the Oudemansiella canarii (Oudemansiella canarii) JZB 2115055.
In the specific primer, the P826-1-2XF and the P826-1-2XR are packaged separately.
In the specific primer, the ratio of the amounts of the P826-1-2XF and the P826-1-2XR may be 1: 1.
Among the specific primers, the specific primer can amplify DNA including DNA molecules shown by SEQ ID No.2 and/or DNA molecules shown by SEQ ID No.3 in a sequence table from genomic DNA of Oudemansiella canarii (Oudemansiella canarii) JZB 2115055.
The invention also provides a reagent or a kit for identifying or assisting in identifying the Oudemansiella canarii (Oudemansiella canarii) JZB2115055, wherein the reagent or the kit comprises the specific primer.
The above-mentioned reagent or kit for identifying or assisting in identifying Oudemansiella canarii (Oudemansiella canarii) JZB2115055 further includes other reagents required for performing a PCR reaction in addition to the primers.
The application of the specific primer in identifying or assisting in identifying the Oudemansiella canarii (Oudemansiella canarii) JZB2115055 or the application in preparing the product for identifying or assisting in identifying the Oudemansiella canarii (Oudemansiella) JZB2115055 also belongs to the protection scope of the invention.
The application of the reagent or the kit in identification or auxiliary identification of the Oudemansiella canarii (Oudemansiella canarii) JZB2115055 also belongs to the protection scope of the invention.
The invention also provides a method for identifying or assisting in identifying Oudemansiella canarii (Oudemansiella canarii) JZB 2115055.
The method for identifying or assisting in identifying the pleurotus eryngii (Oudemansiella canarii) JZB2115055 comprises the steps of using genome DNA of a biological sample to be detected as a template, carrying out PCR amplification by using the P826-1-2XF and the P826-1-2XR to obtain a PCR product, detecting the size of the PCR product, and if the PCR product contains DNA fragments of 537bp and 431bp (or DNA fragments of 537bp and 431 bp), enabling the biological sample to be detected to be the pleurotus eryngii (Oudemansiella canarii) JZB2115055 or contain the pleurotus eryngii (Oudemansiella canarii) JZB 5055, or enabling the biological sample to be detected to be the pleurotus eryngii (Oudemansiella canarii) JZB2115055 or be candidate containing the pleurotus eryngii (ounsiella eryngii) 2115055; if the PCR product does not contain 537bp and 431bp DNA fragments (or 537bp and 431bp DNA fragments), the biological sample to be tested is not Leuconostoc (Oudensiella canarii) JZB2115055 or does not contain Leuconostoc (Oudensiella canarii) JZB2115055, or the biological sample to be tested is not Leuconostoc (Oudensiella canarii) JZB2115055 or is not candidate of Leuconostoc (Oudensiella canarii) JZB 2115055.
The application of the DNA molecule shown in SEQ ID No.3, the DNA molecule shown in SEQ ID No.2 or the DNA molecule shown in SEQ ID No.1 in identification or auxiliary identification of the Oudemansiella (Oudemansiella canarii) JZB2115055 or the application in preparation of products for identification or auxiliary identification of the Oudemansiella (Oudemansiella) JZB2115055 also belongs to the protection scope of the invention.
As described above, the above-described identification or auxiliary identification of the product JZB2115055 of Oudemansiella canarii (Oudemansiella canarii) may be a reagent or kit for identifying or auxiliary identification of the product JZB2115055 of Oudemansiella canarii (Oudemansiella canarii).
In the above application, if the genomic DNA of the biological sample to be tested contains the DNA molecule represented by SEQ ID No.3, the DNA molecule represented by SEQ ID No.2 or the DNA molecule represented by SEQ ID No.1, the biological sample to be tested is Oudemansiella canarii (Oudemansiella canarii) JZB2115055 or contains Oudemansiella canarii (Oudemansiella canarii) JZB2115055, or the biological sample to be tested is selected to be Oudemansiella canarii (Oudemansiella canarii) JZB2115055 or is selected to contain ounsiella canarii (Oudemansiella canarii) JZB 2115055; if the genomic DNA of the biological sample to be detected does not contain the DNA molecule shown in SEQ ID No.3, the DNA molecule shown in SEQ ID No.2 or the DNA molecule shown in SEQ ID No.1, the biological sample to be detected is not pleurotus sajor-caju (Oudensiella canarii) JZB2115055 or does not contain pleurotus sajor-caju (Oudensiella sajori) JZB2115055, or the biological sample candidate to be detected is not pleurotus sajor-caju (Oudensiella sajor) JZB2115055 or the biological sample candidate to be detected does not contain pleurotus sajor-caju (Oudensiella 211211xiii) JZB2115055 5055.
Experiments prove that the DNA molecule shown in SEQ ID No.3, the DNA molecule shown in SEQ ID No.2 or the DNA molecule shown in SEQ ID No.1 is a specific molecular marker of Oudemansiella (Oudemansiella canarii) JZB2115055, and can be used for rapidly identifying Oudemansiella (Oudemansiella canarii) JZB 2115055. The above-mentioned P826-1-2XF and P826-1-2XR are specific PCR primer pairs of Oudemansiella ochracea (Oudemansiella canarii) JZB2115055, and can be used for rapidly identifying Oudemansiella ochracea (Oudemansiella canarii) JZB 2115055.
Deposit description
The strain name is as follows: oudemansiella canarii (Oudemansiella)
The strain number is as follows: JZB2115055
The preservation organization: china general microbiological culture Collection center
The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)
Address: xilu No.1 Hospital No.3 of Beijing market facing Yang district
The preservation date is as follows: 2012, 5 and 28 days
Registration number of the preservation center: CGMCC No.6171
Drawings
FIG. 1 shows the electropherogram of the PCR product of primer P826 against 19 strains of bacteria. Wherein M is 1kb plus DNA ladder, 1 to 19 are strains numbered 1 to 19 in the following embodiments, respectively, and the band in the box is a specific band of Oudemansiella ochracea (Oudemansiella canarii) JZB 2115055.
FIG. 2 is an electrophoretogram of PCR amplification products of primers P826-1-2XR and P826-1-2 XF. Wherein M is 1kb plus DNA ladder, and 1 to 19 are strains numbered 1 to 19 in the following embodiments, respectively.
FIG. 3 is an electropherogram of PCR amplification products of ITS primers. Wherein M is 1kb plus DNA ladder, and 1 to 19 are strains numbered 1 to 19 in the following embodiments, respectively.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The experimental procedures in the following examples are conventional unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The 19 strains referred to in the following examples are as follows:
1. oudemansiella ochroleuca (Oudemansiella canarii) JZB2115055 has been deposited in the China general microbiological culture Collection center (CGMCC) at 28/5/2012, with the collection number of CGMCC No.6171, and has been published in the Chinese patent application with the publication number CN105769938A at 20/7/2016.
2. Oudemansiella canarii JZB2115081 is Oudemansia tropical Oudemansonia (Jun. Nature Culture Collection Center, CFCC for short) in China Center for Culture Collection of microorganisms, which was collected in CFCC at 9.29.2013 and deposited at CFCC as accession number CFCC 52388, and is publicly available from CFCC from the Collection date.
3. The long root mushroom (Oudemansiella radica (Relhan) Singer) with CFCC of Oudemansiella radica (Relhan) Singer has been collected in CFCC at 26.4.2013 with a collection number of CFCC 89567, and is publicly available from CFCC since the collection date.
4. The brown ruffle fimbriatus (Oudemansiella brunneomarginata) is CFCC's brown ruffle fimbriatus (Oudemansiella brunneomarginata lj.n. vasssiljeva), which has been collected in CFCC at 26 months 12 and 2014, with a collection number of CFCC 52389, and is publicly available from CFCC since the collection date.
5. The brown ruffle fimbriatus (Oudemansiella brunneomarginata) is CFCC's brown ruffle fimbriatus (Oudemansiella brunneomarginata lj.n. vasssiljeva), which has been collected in CFCC at 12/27 th 2014 under the collection number CFCC 52390, and is publicly available from CFCC since the collection date.
6. The brown ruffle fimbriatus (Oudemansiella brunneomarginata) is CFCC's brown ruffle fimbriatus (Oudemansiella brunneomarginata lj.n. vasssiljeva), which has been collected in CFCC at 24.12.2014, with a collection number of CFCC 52391, and is publicly available from CFCC since the collection date.
7. Oudemansiella brunneomarginata, an agricultural microorganism center of China Committee for culture Collection of microorganisms (also called China agricultural microorganism culture Collection management center, abbreviated as ACCC, address: No. 12 south street of Guancun in Haishen district, Beijing, institute of agricultural resources and agricultural regionalization, China agricultural academy of sciences, postal code 100081), was collected in ACCC before 2018, and the strain was deposited as ACCC 51130, and was publicly available from ACCC from the date of collection.
8. Glue stickOudemansiella mucida (Oudemansiella mucida) is a CFCC myxoOudemansiella mucida (Schrad.)
) It has been collected in CFCC at 11/2.2013 under the accession number CFCC 52392, and since this collection date the strain was publicly available from CFCC.
9. Oudemansiella radicata (Oudemansiella radica) which is a long root mushroom of CFCC (Oudemansiella radica (Relhan) Singer) has been collected in CFCC at 29.12.2011, with a deposit number of CFCC52393, from which the strain is publicly available from CFCC.
10. Oudemansiella radicata (Oudemansiella radica) which is a long root mushroom of CFCC (Oudemansiella radica (Relhan) Singer) has been collected in CFCC on 27.11.2013 and has a deposit number of CFCC52394, and the strain is publicly available from CFCC on the date of collection.
11. Oudemansiella radicata (Oudemansiella radica) which is a long root mushroom of CFCC (Oudemansiella radica (Relhan) Singer) has been collected in CFCC on 27.11.2013 and has a deposit number of CFCC52395, and the strain is publicly available from CFCC on the date of collection.
12. Oudemansiella radicata (Oudemansiella radica) which is a long root mushroom of CFCC (Oudemansiella radica (Relhan) Singer) has been collected in CFCC at 28.12.2014 under the accession number CFCC52396, and is publicly available from CFCC since the collection date.
13. Oudemansia mucosae (Oudemansiella mucida) is an Asian variant of Oudemansiella mucosae (Oudemansiella mucida (Schrad.) History (n.)) of CFCC and has been collected at CFCC on 12/1 month 2014 under the collection number CFCC 52397, and is publicly available from CFCC on the day of collection.
14. Oudemansia mucosae (Oudemansiella mucida) is an Asian variant of Oudemansiella mucosae (Oudemansiella mucida (Schrad.) History (n.)) of CFCC and has been collected at CFCC on 12/1 month 2014 under the collection number CFCC 52398, and is publicly available from CFCC on the day of collection.
15. Pleurotus citrinopileatus (Oudensiella venosollamata) is CFCC Leontopodium citrinopileatus (Oudensiella venosollamata (Imazeki & Toki) Imazeki & Hongo) which has been collected in CFCC at 27.12.2014 under accession number CFCC 52399, and is publicly available from CFCC since the collection date.
16. Pleurotus citrinopileatus (Oudensiella venosollamata) is CFCC Leontopodium citrinopileatus (Oudensiella venosollamata (Imazeki & Toki) Imazeki & Hongo) which has been collected in CFCC at 26.12.2014 under accession number CFCC 52400, and is publicly available from CFCC since this collection date.
17. Pleurotus citrinopileatus (Oudensiella venosollamata) is CFCC Leontopodium citrinopileatus (Oudensiella venosollamata (Imazeki & Toki) Imazeki & Hongo) which has been collected in CFCC at 12/25 days 2014 under the collection number CFCC 52401, and is publicly available from CFCC since the collection date.
18. Pleurotus citrinopileatus (Oudensiella venosollamata) is CFCC Leontopodium citrinopileatus (Oudensiella venosollamata (Imazeki & Toki) Imazeki & Hongo) which has been collected in CFCC at 27.12.2014 under accession number CFCC 52402, and is publicly available from CFCC since this collection date.
19. Pleurotus citrinopileatus (Oudensiella venosollamata) is CFCC Leontopodium citrinopileatus (Oudensiella venosollamata (Imazeki & Toki) Imazeki & Hongo) which has been collected in CFCC at 27.12.2014 under accession number CFCC 52403, and is publicly available from CFCC since this collection date.
The 19 strains are cultured by adopting a PDA culture medium: peeling 200g of potato, slicing, boiling with boiled water until the potato is not rotten, filtering with gauze, adding 20g of agar into liquid, heating to completely dissolve the agar, adding 20g of glucose, 5g of peptone, 3g of monopotassium phosphate, 1.5g of magnesium sulfate, 110mg of vitamin B and 1L of deionized water, heating the solution, completely dissolving the substances, and then putting the dissolved substances into a conical flask for autoclaving at the sterilization condition of 121 ℃ for 30 min.
Example 1 development of molecular markers for Oudemansiella canarii (Oudemansiella canarii) JZB2115055
Development of specific primer set for Oudemansiella canarii (Oudemansiella canarii) JZB2115055
Genomic DNA of each of the 19 strains was extracted using the DNeasy Plant Mini Kit from Qiagen. The genomic DNA concentration of the 19 strains was adjusted to 50 ng/. mu.l and used as a template, and PCR amplification was performed with 26 primers in Table 1 in sequence, the total volume of PCR reaction was 20. mu.l, containing 10. mu.l of 2 XTaq PCR Master Mix (purchased from the Biotech development center of Yongze Haojia, Beijing), 10. mu. mol/L of primers (1. mu.l each), 50ng of DNA, and the total volume was supplemented to 20. mu.l with sterile double distilled water. And adding the reaction solution into a 0.2ml centrifuge tube according to the volume, uniformly mixing, and putting into a PCR instrument for PCR amplification. PCR amplification conditions: pre-denaturation at 94 ℃ for 4min, cycle parameters of 94 ℃ for 30s, renaturation at a temperature of 5 ℃ lower than the Tm value of the primers for 30s, extension at 72 ℃ for 2min, 30 cycles in total, and extension at 72 ℃ for 7 min. The amplified product was subjected to agarose gel electrophoresis, and a specific fragment of Oudemansiella ochroleuca (Oudemansiella canarii) JZB2115055 was observed.
TABLE 1, 26 primers
The result shows that when the primer P826 is used for amplification, a specific band appears between 700bp and 1000bp of the Oudemansiella canarii (Oudemansiella canarii) JZB2115055, and the specific band is not existed in other 18 strains, which indicates that the P826 is the specific primer of the Oudemansiella fulva (Oudemansiella canarii) JZB2115055 and can be used for rapidly identifying the Oudemansiella fulva (Oudemansiella canarii) JZB 2115055. The specific band was recovered using QIAquick Gel Extraction Kit from Qiagen, and sequenced by the agency of Mimethai and Biotechnology (Beijing) Ltd. The sequencing result shows that the nucleotide sequence of the DNA molecule of the specific strip is sequence 1(SEQ ID No.1) in the sequence table. The DNA molecule shown in the sequence 1 in the sequence table is a specific molecular marker of Oudemansiella canarii (Oudemansiella canarii) JZB2115055, and can be used for quickly identifying Oudemansiella canarii (Oudemansiella canarii) JZB 2115055.
Primer pairs P826-1-2XF (5'-GTGAGCATGTTGAAGCCTTG-3') (SEQ ID No.4) and P826-1-2XR (5'-GCGGAATAAAGCATTTGGTGT-3') (SEQ ID No.5) were designed as SEQ ID No. 1. The genomic DNA concentration of the 19 strains was adjusted to 50 ng/. mu.l and used as a template, and PCR was carried out using primer pairs P826-1-2XF and P826-1-2 XR. The total volume of PCR reaction was 20. mu.l, containing 10. mu.l of 2 XTAQQ PCR Master Mix (purchased from Biotech development center, Yongze Haojia, Beijing), 0.4. mu. mol/L (1. mu.l each of primers), 50ng of DNA, and the total volume was made up to 20. mu.l by adding sterilized double distilled water. And adding the reaction solution into a 0.2ml centrifuge tube according to the volume, uniformly mixing, and putting into a PCR instrument for PCR amplification. Specific primer PCR amplification conditions: pre-denaturation at 95 ℃ for 4min, denaturation at 95 ℃ for 30s, renaturation at 55 ℃ for 30s, extension at 72 ℃ for 2min for 30 cycles, and extension at 72 ℃ for 7 min. The amplified product was subjected to agarose gel electrophoresis, and a specific fragment of Oudemansiella ochroleuca (Oudemansiella canarii) JZB2115055 was observed. The amplification result shows that the Oudemansia fuselacea (Oudemansiella canarii) JZB2115055 (strain with the number of 1) has two DNA bands below 1000bp, the sizes of the two DNA bands are both between 700bp and 400bp, the amplification band of the strain with the numbers of 2-19 is different from the Oudemansia fuselacea (Oudemansiella canarii) JZB2115055, and no DNA band or the amplification band is between 5000bp and 2000 bp. The two specific bands of Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 are recovered and subjected to sequence determination, and the sequencing result shows that the nucleotide sequences of the DNA molecules of the two specific bands are respectively sequence 2(SEQ ID No.2) in the sequence table and sequence 3(SEQ ID No.3) in the sequence table, and the lengths are 537bp and 431bp respectively. The DNA molecule shown in the sequence 2 in the sequence table and the DNA molecule shown in the sequence 3 in the sequence table are specific molecular markers of Oudemansiella ochracea (Oudemansiella canarii) JZB2115055, and can be used for quickly identifying Oudemansiella ochracea (Oudemansiella canarii) JZB 2115055. Specific primer pairs P826-1-2XF and P826-1-2XR are adopted to accurately and rapidly identify the Oudemansiella ochracea (Oudemansiella canarii) JZB 2115055.
Meanwhile, the genomic DNA of the strains numbered 1-19 is used as a template and an ITS primer is used for amplification (figure 3), all strains have correct ITS sequences, and the extracted genomic DNA is proved to have good effect. The ITS primers are ITS4 and ITS 5. The sequence of ITS4 with sequence 5'-TCCTCCGCTTATTGATATGC-3', ITS5 is 5'-GGAAGTAAAAGTCGTAACAAGG-3'.
<110> agriculture and forestry academy of sciences of Beijing City
<120> molecular marker of oudemansiella lutescens JZB2115055 and application thereof
<160> 5
<170> PatentIn version 3.5
<210> 1
<211> 753
<212> DNA
<213> Oudemansiella canarii (Oudemansiella)
<400> 1
acacacacac acacacccat ctacatacgc agctgtattt actatcctaa tccaggttcg 60
tttgtttggg ctatgggtag ggcgtggccg gccaccagga gtattttaaa tattgtgatg 120
tgaatgcact agtgtactga ttcatgttat tgtggcgtaa ttttgttatc gatgggtgac 180
tatgaataag tgcccggggt gagcatgttg aagccttgct gtttctttac gtgtaagttc 240
gtgttggcgc tattttttat agtccgctgg tatgtatact tctttcattg gtgacctcat 300
gttttcgtta tgtagaatgt taggcatgcg acgggggcgg aaggtgaaag cactgacagg 360
tgattcttgt gtggcgccga gtgagaatat atgtgcttgc cttgggagta tggcaaggcc 420
tattagggag agggccggaa gtacttaatc agctagagca ggtctgagta aggccgcctt 480
gcaagtacta ctcaacgtct acacagatgt gcttgtttag tctaacggct caagcgtgtg 540
taccattgtc tggtaggcta acccgtactc tttttttgtt ctgaagcttc ctgatgctgg 600
atagacgctc atcaatacta atctacagag ttatagccta tgccagtgaa tgcgccacgg 660
atgagatcgc gagggctgtg tgcccagtgg gggctgaatt catcccaagt atcgacacca 720
aatgctttat tccgctggtg tgtgtgtgtg tgt 753
<210> 2
<211> 537
<212> DNA
<213> Oudemansiella canarii (Oudemansiella)
<400> 2
gtgagcatgt tgaagccttg ctgtttcttt acgtgtaagt tcgtgttggc gctatttttt 60
atagtccgct ggtatgtata cttctttcat tggtgacctc atgttttcgt tatgtagaat 120
gttaggcatg cgacgggggc ggaaggtgaa agcactgaca ggtgattctt gtgtggcgcc 180
gagtgagaat atatgtgctt gccttgggag tatggcaagg cctattaggg agagggccgg 240
aagtacttaa tcagctagag caggtctgag taaggccgcc ttgcaagtac tactcaacgt 300
ctacacagat gtgcttgttt agtctaacgg ctcaagcgtg tgtaccattg tctggtaggc 360
taacccgtac tctttttttg ttctgaagct tcctgatgct ggatagacgc tcatcaatac 420
taatctacag agttatagcc tatgccagtg aatgcgccac ggatgagatc gcgagggctg 480
tgtgcccagt gggggctgaa ttcatcccaa gtatcgacac caaatgcttt attccgc 537
<210> 3
<211> 431
<212> DNA
<213> Oudemansiella canarii (Oudemansiella)
<400> 3
gtgagcatgt tgaagccttg ctgtttcttt acgtgtaagt tcgtgttggc ggtatttttt 60
atagtccgct ggtatgtata cttctttcat tggtgacctc atgttttcgt tatgtagaat 120
gttaggcatg cgacgggggc ggaaggtgaa agcactggaa tgtgattctt gtgtggcgcc 180
gagtgaaaat atatgtgctt gcgttggtgt acgaggccga ggagagcatc gctacgcggg 240
gcatgtttaa gtaaggccta tctacgtctg ttgatgttac acggatgtgc ttgtttagag 300
tctaacggct caagtgtatg tgtgatggtc tggtggctag ctcatcagta gttgtgccag 360
tgattgcgcc acaggtgaga tcgcgagggc tgaattcatc ccaaatatcg acaccaaatg 420
ctttattccg c 431
<210> 4
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
gtgagcatgt tgaagccttg 20
<210> 5
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 5
gcggaataaa gcatttggtg t 21
Claims (10)
1. Specific primers of Oudemansiella ochracea (Oudemansiella canarii) JZB2115055, wherein the specific primers can be used for amplifying DNA molecules shown as SEQ ID No.2 and/or DNA molecules shown as SEQ ID No.3 in a sequence table from genomic DNA of Oudemansiella ochracea (Oudemansiella canarii) JZB 2115055; the preservation number of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 6171.
2. The specific primer of claim 1, wherein: the specific primer consists of P826-1-2XF and P826-1-2 XR; the P826-1-2XF is a single-stranded DNA molecule shown in SEQ ID No. 4; the P826-1-2XR is a single-stranded DNA molecule shown in SEQ ID No. 5.
3. The specific primer of claim 1, wherein: the specific primer can amplify DNA molecules shown in SEQ ID No.1 from genomic DNA of Oudemansiella canarii (Oudemannsiella canarii) JZB 2115055.
4. A reagent or kit for identifying or assisting in identifying Oudemansiella canarii (Oudemansiella canarii) JZB2115055, comprising a specific primer according to any one of claims 1-3; the preservation number of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 6171.
5. Use of the specific primers of any one of claims 1 to 3 for identifying or assisting in identifying Oudemansiella canarii (Oudemansiella canarii) JZB 2115055; the preservation number of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 6171.
6. Use of the specific primers of any one of claims 1 to 3 for the preparation of a product for identifying or assisting in identifying the Oudemansiella canarii (Oudemansiella canarii) JZB 2115055; the preservation number of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 6171.
7. Use of the reagent or kit of claim 4 for identifying or assisting in identifying Oudemansiella canarii (Oudemansiella canarii) JZB 2115055; the preservation number of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 6171.
8. A method for identifying or assisting in identifying pleurotus sajor-caju (Oudemansiella canarii) JZB2115055, comprising performing PCR amplification using genomic DNA of a biological sample to be tested as a template by using specific primers as claimed in claim 2 to obtain a PCR product, detecting the size of the PCR product, and if the PCR product contains DNA fragments of 537bp and 431bp, determining that the biological sample to be tested is pleurotus sajor-caju (Oudemansiella sajor) JZB2115055 or contains pleurotus sajor-caju (Oudemansiella sajor) JZB2115055, or the biological sample to be tested is selected from pleurotus sajor-cajor (Oudemansiella canarii) 362115055 or is selected from among fijor-cajor-caju (ounsiella sajor-caju-cajor-cajo (ounsiella canarii) JZB 2115055; if the PCR product does not contain DNA fragments of 537bp and 431bp, the biological sample to be tested is not Oudemansiella fusca (Oudemansiella canarii) JZB2115055 or does not contain Oudemansiella fusca (Oudemansiella canarii) JZB2115055, or the biological sample to be tested is not Oudemansiella fusca (Oudemansiella canarii) JZB2115055 or is not candidate containing Oudemansiella fusca (Oudemansiella canarii) JZB 2115055; the preservation number of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 6171.
The DNA molecule shown in SEQ ID No.2, the DNA molecule shown in SEQ ID No.3 or the DNA molecule shown in SEQ ID No. 1.
The use of the DNA molecule shown in SEQ ID No.2, the DNA molecule shown in SEQ ID No.3 or the DNA molecule shown in SEQ ID No.1 for identifying or assisting in identifying the Leucocyte (Oudensiella canarii) JZB2115055 or for preparing products for identifying or assisting in identifying the Leucocyte (Oudensiella canarii) JZB 2115055; the preservation number of the Oudemansiella ochracea (Oudemansiella canarii) JZB2115055 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 6171.
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