CN110241249A - The primer and method of agaricus bisporus Wet bull pathogen in a kind of quick detection earthing - Google Patents
The primer and method of agaricus bisporus Wet bull pathogen in a kind of quick detection earthing Download PDFInfo
- Publication number
- CN110241249A CN110241249A CN201910652794.2A CN201910652794A CN110241249A CN 110241249 A CN110241249 A CN 110241249A CN 201910652794 A CN201910652794 A CN 201910652794A CN 110241249 A CN110241249 A CN 110241249A
- Authority
- CN
- China
- Prior art keywords
- agaricus bisporus
- primer
- pathogen
- soil
- dna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- 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
Abstract
The invention discloses the primer and method of agaricus bisporus Wet bull pathogen in a kind of quickly detection earthing, withGH63Gene is as specific molecular marker, design the specific primer of amplification Mycogone perniciosa, it is quickly detected before disease has not occurred by the method for PCR amplification in agaricus bisporus soil covering layer with the presence or absence of Mycogone perniciosa spore, play the role of positive for the prevention and treatment of agaricus bisporus Wet bull evil, also solves the deficiency of conventional method poor in timeliness.
Description
Technical field
The invention belongs to the pathogenic microorganism examination technical fields, and in particular to a kind of quickly detect has in agaricus bisporus soil covering
The primer and method of evil Wet bull pathogen.
Background technique
Agaricus bisporus (Agaricus bisporus) is the edible mushroom cultivated extensively in the world, but during the growth process easily
Agaricus bisporus Wet bull occurs, often results in serious economic loss, influences the economic benefit of agaricus bisporus industry.
Agaricus bisporus Wet bull is the important disease during cultivation of agaricus bisporus, often affects the production of agaricus bisporus
Industry causes serious economic loss, its pathogen Mycogone perniciosa (Hypomyces perniciosus) is common soil
Fungi, route of transmission is mainly propagated by chlamydospore, and chlamydospore is primarily present in earthing, its adaptability is very
By force, the waste around mushroom house and soil at the disease it is initial it is main infect source, the conidium of Mycogone perniciosa,
Chlamydospore and mycelium have pathogenic ability, and Spores amount also will affect the disease incidence of Wet bull in earthing.
The identification method of currently used agaricus bisporus Mycogone perniciosa pathogen mainly passes through acquisition agaricus bisporus and covers
Earth sample carries out microorganism and is separately cultured, later by constantly purifying, obtains the pure culture of pathogen, then carry out form
Observation, and extracts fungal DNA, carries out ITS amplification, sequence is compared after sequencing after identifying judge its for Mycogone perniciosa,
The method is often time-consuming and laborious, cumbersome, and program is complicated, obtains having had already passed by optimal Control stage when result,
So by extracting the fungi total DNA in soil, using the specific molecular marker of Mycogone perniciosa, with round pcr (polymerization
Enzyme chain reaction) to come to judge that the presence or absence of pathogen is most important what disease had not occurred, this method simplicity saves trouble, program letter
It is single.
Summary of the invention
In view of the above-mentioned problems, the object of the present invention is to provide in a kind of based on PCR technology qualitative detection agaricus bisporus soil covering
The primer and method of Mycogone perniciosa pathogen, the method can be quickly detected harmful wart in agaricus bisporus soil covering material
The mould spore of spore finds disease before disease generation, is prevented and treated in advance that the disease early warning for agaricus bisporus Wet bull evil mentions
For technical support.
To achieve the above object, The technical solution adopted by the invention is as follows:
According to GH63 gene design primer GH63-F (SEQ ID NO.1) and GH63-R (SEQ ID NO.2), it is used as quick
Detect the primer of agaricus bisporus Wet bull pathogen in earthing.
A kind of method of agaricus bisporus Wet bull pathogen in detection earthing, comprising the following steps:
(1) agaricus bisporus soil covering is acquired, soil total DNA is extracted；
(2) using soil total DNA as template, primer GH63-F (SEQ ID NO.1) and GH63-R (SEQ ID NO.2) are used
Carry out PCR amplification；
(3) amplified production is subjected to nucleic acid electrophoresis detection, if detecting the amplified band of 326bp in pedotheque,
Illustrate there are harmful Wet bull fungal pathogens in earthing.Preferably, agaricus bisporus soil covering exists in above-mentioned detection method step (1)
Soil total DNA is extracted again after expanding culture in PDA liquid medium.
Preferably, in above-mentioned detection method PCR program be 95 DEG C of initial denaturations 5min, 95 DEG C of denaturation 30s, 57 DEG C of annealing 30s,
72 DEG C of extension 1min, 35 circulations, 72 DEG C, 10min, 4 DEG C preservations.
Compared with the prior art, the device have the advantages that it is as follows:
1, the present invention finds GH63 gene order according to the annotation information of Mycogone perniciosa genome, closer edge species
GH63 homologous gene designs the specific primer of amplification Mycogone perniciosa, using the primer using agaricus bisporus soil covering DNA as mould
Plate obtains the Mycogone perniciosa specific product that length is 326bp.
2, quickly being detected by the method for PCR amplification whether there is Mycogone perniciosa spore in agaricus bisporus soil covering layer, into
Row is prevented and treated in advance, plays the role of positive for the prevention and treatment of agaricus bisporus Wet bull evil, also solves conventional method timeliness
The deficiency of difference.
Detailed description of the invention
Fig. 1 is the electrophoretogram in 1 step of the embodiment of the present invention (1) after GH63 primer amplification.M:BM2000Marker, swimming lane
1-9 is followed successively by Trichoderma atroviride, Trichoderma harzianum, Trichoderma viride, square spore trichoderma, long shoot trichoderma, Cladobotryum cubitense,
Cladobotryum protrusum, Cladobotryum sp., Mycogone perniciosa, swimming lane 10 are the negative control without DNA.
Fig. 2 is primer sensitivity test figure in 1 step of the embodiment of the present invention (2).M:BM2000Marker, swimming lane 1-12 are
In PCR reaction system Mycogone perniciosa total DNA content be followed successively by 100,50,25,10,5,2.5,1,0.5,0.25,0.1,0.05,
0ng。
Fig. 3 is the electrophoretogram in 1 step of the embodiment of the present invention (4) after GH63 primer amplification soil DNA.M:
BM2000Marker, swimming lane 1-3: being soil DNA sample, Mycogone perniciosa DNA, negative control respectively.
Fig. 4 is the ITS primer amplification figure of 2 step of the embodiment of the present invention (1).M:BM2000Marker, swimming lane 1-12 difference
It is the soil DNA sample of random acquisition after being vaccinated with Mycogone perniciosa spore suspension, swimming lane 13 is Mycogone perniciosa DNA, swimming lane
14 be negative control.
Fig. 5 is the electrophoretogram after the GH63 primer amplification of 2 step of the embodiment of the present invention (2).M:BM2000Marker, swimming lane
1-12 is the soil DNA sample of random acquisition after being vaccinated with various concentration Mycogone perniciosa spore suspension respectively, and swimming lane 13 is to have
The evil mould DNA of wart spore, swimming lane 14 are negative control.
Specific embodiment
Embodiment 1
Specific primer design and detection method:
(1) according to the fungal gene group annotation information of pathogen Mycogone perniciosa, some species identifications that are usually used in are found
With the conservative gene of species systematic growth research, these gene orders are made comparisons with the orthologous gene sequence of nearly edge species
Analysis, using the sequence difference between homologous gene, using 5.0 software Design primers of Primer Primier, with designed
Primer pair Mycogone perniciosa bacterial strain, 5 trichoderma strains, 3 spider web disease cause of disease bacteria strains DNA make PCR amplification, only basis
The primer GH63-F (SEQ ID NO.1) and GH63-R (SEQ ID NO.2) of GH63 gene design are expanded in Mycogone perniciosa
Specific band out, and in trichoderma and spider web disease pathogen without amplified production.
PCR reaction system is totally 20 μ L, wherein (it is limited that biotechnology is only praised in Nanjing promise to 2 × Taq Plus Master Mix
Company) 10 μ L, upstream and downstream primer (10 μm of ol/L) each 0.5 μ L, 1 μ L of DNA profiling add ddH2O supplies total volume to 20 μ L；
PCR program is 95 DEG C of initial denaturation, 5min, denaturation: 95 DEG C, 30s, annealing: 57 DEG C, 30s, extend: 72 DEG C, 1min,
35 circulations, 72 DEG C, 10min, 4 DEG C preservations.
(2) sensitivity is tested: add 100 in each PCR reaction system respectively, 50,25,10,5,2.5,1,0.5,0.25,
0.1, the Mycogone perniciosa bacterial strain of 0.05,0ng DNA carry out primer sensitivity test, the results showed that, GH63 primer it is sensitive
Property is very high, and only the DNA profiling of 0.05ng can amplify purpose band.
(3) applicant samples from the Hubei garden Ya Jun Biotechnology Co., Ltd base, obtains agaricus bisporus soil covering, weighs 8g
Fresh soil cultivates 2d into PDA liquid medium, carries out the numerous culture of expansion of pathogen.
(4) E.Z.N.A. is utilizedSoil DNA Kit (is purchased from Omega Bio-tek company), and it is total to extract the numerous rear soil of expansion
DNA, and total DNA is dissolved in ddH2In O；With GH63-F and GH63-R respectively as upstream and downstream primer, using soil total DNA as mould
Plate carries out purpose band amplification；
PCR reacts laggard row agarose gel electrophoresis, the single PCR product of 326bp is detected in pedotheque, by PCR
Product is sent to sequencing company sequencing, and nucleotide sequence such as SEQ ID NO.3 determines that GH63-F and GH63-R amplification pedotheque obtains
The product obtained is the single product of specificity, and identical with the sequence of Mycogone perniciosa GH63 gene.
Embodiment 2
It is 10 that Mycogone perniciosa spore suspension concentration gradient, which is arranged,2、104、106A/mL is with spore dilution (water)
Control respectively handles 3 repetitions, is inoculated into the earthing surface of rigid earthing bedstead, each cell 30mL, uses plastics between each processing
Plate separates, totally 12 cells, area equation (1.4 × 1.5m of each cell2)。
(1) in test area grab sample, after cultivating 2d in PDA liquid medium, the numerous culture of expansion of pathogen is carried out,
Soil DNA is extracted, ITS amplification is carried out, each sample can amplify a plurality of band, illustrate that the soil DNA extracted can be used for
PCR amplification.
(2) soil DNA sample is subjected to PCR with primer GH63-F (SEQ ID NO.1) and GH63-R (SEQ ID NO.2)
Amplification, all samples all obtain the single amplified production with Mycogone perniciosa DNA same size, and with ddH2O is negative right
According to when there is no product.
Pedotheque is extracted into total DNA again after cultivation, carries out PCR with the GH63 gene-specific primer of Mycogone perniciosa
Amplification, can obtain the single amplified production with same size in Mycogone perniciosa DNA, illustrate that this method can be examined specifically
It measures in agaricus bisporus soil covering with the presence or absence of disease fungus Mycogone perniciosa.
Sequence table
<110>Hua Zhong Agriculture University
<120>a kind of primer and method for quickly detecting agaricus bisporus Wet bull pathogen in earthing
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 23
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
tctgcatagg atcggtcaat aag 23
<210> 2
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
ccaccaccgt ggatgatata c 21
<210> 3
<211> 326
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
tctgcatagg atcggtcaat aagctggtga ccgtggaagt atcccacgcc ccctagcaag 60
ttggagaaca tgctcttgcc aaacttctgg tacttttccg ccgtataagg tgccttgaga 120
tcaaagacgc tggaaaaccg ctttccaaat gcttcagata cttccttcac ctctcgagtc 180
acgtcctcgg tcgtcagctc cctacctgca gatgccgacg agaagatgac gtcgaactcg 240
aacgacccct caaacacttt ctggatgacg tgggcatttc cacccccagg cttgggatct 300
attcggtata tcatccacgg tggtgg 326
Claims (4)
1. quickly detecting the primer of agaricus bisporus Wet bull pathogen in earthing, feature exists, primer sequence such as SEQ ID
Shown in NO.1 and SEQ ID NO.2.
2. using the method for agaricus bisporus Wet bull pathogen in primer detection earthing described in claim 1, feature exists
In, comprising the following steps:
(1) agaricus bisporus soil covering is acquired, soil total DNA is extracted；
(2) using soil total DNA as template, PCR amplification is carried out using primer described in claim 1；
(3) amplified production is subjected to nucleic acid electrophoresis detection.
3. according to the method described in claim 2, it is characterized in that, agaricus bisporus soil covering is in PDA liquid medium in step (1)
Soil total DNA is extracted again after middle expansion culture.
4. according to the method described in claim 2, it is characterized in that, PCR program be 95 DEG C of initial denaturations 5min, 95 DEG C of denaturation 30s,
57 DEG C of annealing 30s, 72 DEG C of extension 1min, 35 circulations, 72 DEG C of extension 10min, 4 DEG C save.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910652794.2A CN110241249B (en) | 2019-07-18 | 2019-07-18 | Primer and method for rapidly detecting pathogen of agaricus bisporus wart spongiform in covering soil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910652794.2A CN110241249B (en) | 2019-07-18 | 2019-07-18 | Primer and method for rapidly detecting pathogen of agaricus bisporus wart spongiform in covering soil |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110241249A true CN110241249A (en) | 2019-09-17 |
| CN110241249B CN110241249B (en) | 2021-03-05 |
Family
ID=67892882
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910652794.2A Active CN110241249B (en) | 2019-07-18 | 2019-07-18 | Primer and method for rapidly detecting pathogen of agaricus bisporus wart spongiform in covering soil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110241249B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111100940A (en) * | 2020-01-21 | 2020-05-05 | 华中农业大学 | Method for rapidly detecting pseudomonas in needle mushroom culture material |
| CN111500760A (en) * | 2020-05-09 | 2020-08-07 | 福建省农业科学院植物保护研究所 | Molecular detection primer for rapidly identifying verruca rosea and application thereof |
| CN112522428A (en) * | 2020-12-10 | 2021-03-19 | 华中农业大学 | Method for rapidly detecting Arthrobacter attheifer in needle mushroom culture material |
| CN111549166B (en) * | 2020-05-09 | 2022-05-20 | 福建省农业科学院植物保护研究所 | Molecular detection primer for new species of verrucaria sporogenes and application |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012021883A2 (en) * | 2010-08-13 | 2012-02-16 | Dyadic International, Inc. | Novel fungal enzymes |
| CN102703362A (en) * | 2012-06-21 | 2012-10-03 | 福建农林大学 | Streptomyces fujianensis sp and DNA identification method |
| WO2013034939A2 (en) * | 2011-09-08 | 2013-03-14 | Szegedi Tudományegyetem | Active agents against pseudomonas species causing rotting diseases in mushroom production, their use and compositions containing them |
| CN105462864A (en) * | 2015-12-25 | 2016-04-06 | 青岛蔚蓝生物集团有限公司 | Aspergillus niger mutant strain capable of producing alpha-glucosaccharase |
| CN105734132A (en) * | 2016-03-18 | 2016-07-06 | 福建省农业科学院植物保护研究所 | Agaricus bisporus Monilinia fructicola molecular detection primer and quick detection method |
| CN107208098A (en) * | 2014-12-16 | 2017-09-26 | 美国陶氏益农公司 | For controlling coleopteran pest chromatin remodeling gene parental generation RNAI to suppress |
| CN107257853A (en) * | 2014-12-29 | 2017-10-17 | 尹特荣生物科技株式会社 | New enterohemorrhagic escherichia coli phage E sc CHP 1 and its purposes for suppressing enterohemorrhagic escherichia coli propagation |
-
2019
- 2019-07-18 CN CN201910652794.2A patent/CN110241249B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012021883A2 (en) * | 2010-08-13 | 2012-02-16 | Dyadic International, Inc. | Novel fungal enzymes |
| WO2013034939A2 (en) * | 2011-09-08 | 2013-03-14 | Szegedi Tudományegyetem | Active agents against pseudomonas species causing rotting diseases in mushroom production, their use and compositions containing them |
| CN102703362A (en) * | 2012-06-21 | 2012-10-03 | 福建农林大学 | Streptomyces fujianensis sp and DNA identification method |
| CN107208098A (en) * | 2014-12-16 | 2017-09-26 | 美国陶氏益农公司 | For controlling coleopteran pest chromatin remodeling gene parental generation RNAI to suppress |
| CN107257853A (en) * | 2014-12-29 | 2017-10-17 | 尹特荣生物科技株式会社 | New enterohemorrhagic escherichia coli phage E sc CHP 1 and its purposes for suppressing enterohemorrhagic escherichia coli propagation |
| CN105462864A (en) * | 2015-12-25 | 2016-04-06 | 青岛蔚蓝生物集团有限公司 | Aspergillus niger mutant strain capable of producing alpha-glucosaccharase |
| CN105734132A (en) * | 2016-03-18 | 2016-07-06 | 福建省农业科学院植物保护研究所 | Agaricus bisporus Monilinia fructicola molecular detection primer and quick detection method |
Non-Patent Citations (4)
| Title |
|---|
| DAN LI等: "Genome Analysis of Hypomyces perniciosus, the Causal Agent of Wet Bubble Disease of Button Mushroom ( Agaricus bisporus)", 《GENES》 * |
| KUBICEK,C.P.等: "Trichoderma atroviride IMI 206040 glycoside hydrolase family 63 protein mRNA", 《GENBANK》 * |
| MICHAEL A. KERTESZ等: "Compost bacteria and fungi that influence growth and development of Agaricus bisporus and other commercial mushrooms", 《APPLIED MICROBIOLOGY AND BIOTECHNOLOGY》 * |
| 张春兰: "对双孢蘑菇褐腐病致病菌有害疣孢霉的研究", 《中国优秀硕士学位论文全文数据库农业科技辑（月刊）》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111100940A (en) * | 2020-01-21 | 2020-05-05 | 华中农业大学 | Method for rapidly detecting pseudomonas in needle mushroom culture material |
| CN111500760A (en) * | 2020-05-09 | 2020-08-07 | 福建省农业科学院植物保护研究所 | Molecular detection primer for rapidly identifying verruca rosea and application thereof |
| CN111549166B (en) * | 2020-05-09 | 2022-05-20 | 福建省农业科学院植物保护研究所 | Molecular detection primer for new species of verrucaria sporogenes and application |
| CN111500760B (en) * | 2020-05-09 | 2022-05-20 | 福建省农业科学院植物保护研究所 | Molecular detection primer for rapidly identifying verruca rubra and application |
| CN112522428A (en) * | 2020-12-10 | 2021-03-19 | 华中农业大学 | Method for rapidly detecting Arthrobacter attheifer in needle mushroom culture material |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110241249B (en) | 2021-03-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110241249A (en) | The primer and method of agaricus bisporus Wet bull pathogen in a kind of quick detection earthing | |
| CN106282397A (en) | Identify mating type method and the primer pair of four kinds in black Morchella esculenta (L.) Pers monoid | |
| CN108893557A (en) | A kind of method of three kinds of wheat rhizome portion diseases of quick detection | |
| CN105256060A (en) | PCR detection primer and method for anoectochilus roxburghii colletotrichum orbiculare | |
| CN105177148A (en) | Dual PCR primer for detecting two types of pathogens of grape canker at same time and application of dual PCR primer | |
| CN104946760B (en) | A kind of method and dedicated kit for detecting Pythium inflatum | |
| CN106282396A (en) | Identify method and the special primer pair of ladder rib Morchella esculenta (L.) Pers mating type | |
| CN103184281B (en) | Method for identifying or assisting in identifying mating types of Lepista sordid protoplast monokaryons and special primer pairs IS-873 thereof | |
| CN111100940B (en) | Method for rapidly detecting pseudomonas in needle mushroom culture material | |
| CN112662804B (en) | Primer group, kit and method for detecting pathogenic variation of avirulence gene AvrPi9 of rice blast | |
| CN112176080B (en) | Nested PCR primer group, kit and detection method for specifically detecting purple sisal leaf roll disease phytoplasma | |
| JP2006238838A (en) | Method for assaying existence of bacterium | |
| CN103276070A (en) | Method for identifying mating types of lepista sordida protoplasted monokaryons and special primer pair IS-879b thereof | |
| CN105886643A (en) | PCR (polymerase chain reaction) detection primers and method for Aspergillus fumigatus | |
| CN106978505B (en) | Primer pair for detecting pedunculosis solani and application thereof | |
| Chehri | Molecular phylogeny of the Fusarium solani species complex (FSSC) isolated from soils in Iran | |
| CN106244721B (en) | A kind of capsicum stain anthrax bacteria molecular detection primer and detection method | |
| CN103305606B (en) | Method for identifying mating type of protoplasted monokaryon of lepista sordida and special primer pair SR-6*4 thereof | |
| CN103160593B (en) | Specific detection method of phytophthora sojae avirulence gene (Avrlc ) | |
| CN116144831B (en) | Primer combination for identifying mating type tremella aurantialba mononucleosome and identification method | |
| CN113897456B (en) | Real-time fluorescence PCR detection primer probe combination and detection kit for four fir anthracnose pathogens and application of kit | |
| CN103194535B (en) | Method for identification or auxiliary identification of mating type of protoplast monokaryon of Lepista sordid and special primer pair SR-5*16 used therein | |
| CN103184282B (en) | Method and special primer SR-5*13 for differentiating and auxiliarily differentiating mating types of bioplast monokaryons of lepista sordida (Fr) sings | |
| CN105463107A (en) | Primer pair AWF-AWR for identifying pantoea beijingensis | |
| CN103233081B (en) | Method for identifying mating types of Lepista sordida protoplast monokaryons and special primer pair SR-6*6 therefor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |