CN116426394A - 一种玉龙杓兰共生真菌及应用 - Google Patents
一种玉龙杓兰共生真菌及应用 Download PDFInfo
- Publication number
- CN116426394A CN116426394A CN202310611837.9A CN202310611837A CN116426394A CN 116426394 A CN116426394 A CN 116426394A CN 202310611837 A CN202310611837 A CN 202310611837A CN 116426394 A CN116426394 A CN 116426394A
- Authority
- CN
- China
- Prior art keywords
- biaolan
- symbiotic
- fungus
- culture
- jade
- 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
- 241000233866 Fungi Species 0.000 title claims abstract description 67
- 239000010977 jade Substances 0.000 title claims abstract description 41
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 26
- 241000894006 Bacteria Species 0.000 claims abstract description 25
- 238000004321 preservation Methods 0.000 claims abstract description 22
- 241001328209 Magnibotryascoma Species 0.000 claims abstract description 13
- 239000000284 extract Substances 0.000 claims abstract description 11
- 239000001963 growth medium Substances 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- 241001516485 Cypripedium Species 0.000 claims description 18
- 239000008223 sterile water Substances 0.000 claims description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 238000012258 culturing Methods 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 12
- 239000000287 crude extract Substances 0.000 claims description 11
- 239000007787 solid Substances 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- 229960000723 ampicillin Drugs 0.000 claims description 9
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 238000000605 extraction Methods 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 238000002791 soaking Methods 0.000 claims description 7
- 230000001954 sterilising effect Effects 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 6
- 241000235349 Ascomycota Species 0.000 claims description 5
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 5
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- 238000010992 reflux Methods 0.000 claims description 5
- 238000007790 scraping Methods 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 230000006037 cell lysis Effects 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 4
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- 244000061456 Solanum tuberosum Species 0.000 claims description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 229930027917 kanamycin Natural products 0.000 claims description 3
- 229960000318 kanamycin Drugs 0.000 claims description 3
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 claims description 3
- 229930182823 kanamycin A Natural products 0.000 claims description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 3
- 241000723418 Carya Species 0.000 claims description 2
- 229940124350 antibacterial drug Drugs 0.000 claims description 2
- 239000008188 pellet Substances 0.000 claims description 2
- 239000001103 potassium chloride Substances 0.000 claims description 2
- 235000011164 potassium chloride Nutrition 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 239000004317 sodium nitrate Substances 0.000 claims description 2
- 235000010344 sodium nitrate Nutrition 0.000 claims description 2
- JZRWCGZRTZMZEH-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 2
- 230000000813 microbial effect Effects 0.000 claims 2
- 238000007605 air drying Methods 0.000 claims 1
- 241000194103 Bacillus pumilus Species 0.000 abstract description 7
- 244000063299 Bacillus subtilis Species 0.000 abstract description 7
- 235000014469 Bacillus subtilis Nutrition 0.000 abstract description 7
- 241000191938 Micrococcus luteus Species 0.000 abstract description 7
- 241000607762 Shigella flexneri Species 0.000 abstract description 7
- 241000193985 Streptococcus agalactiae Species 0.000 abstract description 7
- 241000193422 Bacillus lentus Species 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 16
- 239000000523 sample Substances 0.000 description 16
- 239000000243 solution Substances 0.000 description 16
- 241000196324 Embryophyta Species 0.000 description 15
- 230000009089 cytolysis Effects 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 7
- 238000001514 detection method Methods 0.000 description 5
- 241000193830 Bacillus <bacterium> Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 244000052616 bacterial pathogen Species 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 3
- 230000002538 fungal effect Effects 0.000 description 3
- 238000000197 pyrolysis Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 108091023242 Internal transcribed spacer Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 241000751139 Beauveria bassiana Species 0.000 description 1
- 241001176652 Cypripedium forrestii Species 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000223250 Metarhizium anisopliae Species 0.000 description 1
- 244000262613 Michelia alba Species 0.000 description 1
- 235000003415 Michelia alba Nutrition 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 241001092500 Photinia x fraseri Species 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 229930003270 Vitamin B Natural products 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- BKHZIBWEHPHYAI-UHFFFAOYSA-N chloroform;3-methylbutan-1-ol Chemical compound ClC(Cl)Cl.CC(C)CCO BKHZIBWEHPHYAI-UHFFFAOYSA-N 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 239000002274 desiccant Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000003517 fume Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000013595 supernatant sample Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Botany (AREA)
- Animal Behavior & Ethology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Alternative & Traditional Medicine (AREA)
- Medical Informatics (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
本发明属于微生物技术领域,特别涉及一种玉龙杓兰共生真菌及应用,该真菌的保藏名称为昆明巨丛子囊(Magnibotryascoma kunmingense)YAFEF024,保藏于中国典型培养物保藏中心,保藏地址为中国.武汉.武汉大学,保藏编号为CCTCC NO:M 2022613,保藏时间为2022年05月12日。本发明提供一种玉龙杓兰共生真菌及应用,其菌丝体提取物对缓慢芽孢杆菌、无乳链球菌、短小芽孢杆菌、福氏志贺氏菌、枯草芽孢杆菌、藤黄微球菌等细菌具有较好的抗细菌活性。
Description
技术领域
本发明属于微生物技术领域,特别涉及一种玉龙杓兰共生真菌及应用。
背景技术
共生真菌(Plant symbiotic fungi)是植物生活史的特定阶段或全部阶段寄生在植物组织内,而不引起植物产生有害症状的微生物,包括菌根真菌以及生活在植物表面的内生真菌和潜伏性病原菌。植物共生真菌能产生一些次级代谢物质,具有抗菌、抗氧化、抗肿瘤、促进作物生长以及提高作物抗病性等特性。
通常将能与植物建立互惠共生关系的真菌统称为植物互惠共生真菌,主要包括外生菌根真菌、丛枝菌根真菌、兰科菌根真菌和欧石楠菌根真菌、暗隔内生真菌、印度梨形孢、木霉、白僵菌和绿僵菌等。这些共生真菌可以在健康植物组织内定殖生活却不会对植物产生明显的伤害,还能促进植物生长,提高植物对抗胁迫能力。因此从植物共生真菌中寻找潜在的生防菌是一个非常好的选择,该领域已受到越来越多生物学家及生态学家们的关注。
玉龙杓兰(Cypripedium forrestii)是杓兰属植物,由植物学家George Forrest于1913年在云南省丽江地区采集并命名,为中国特有濒危植物。分布于中国云南西北部(丽江、中甸),生于海拔3500米的松林下、灌木丛生的坡地或开旷林地上。而目前关于玉龙杓兰的相关报道较少。
发明内容
本发明提供一种玉龙杓兰共生真菌及应用,其菌丝体提取物对缓慢芽孢杆菌、无乳链球菌、短小芽孢杆菌、福氏志贺氏菌、枯草芽孢杆菌、藤黄微球菌等细菌具有较好的抗细菌活性。
本发明通过下述技术方案实现:
一方面,本申请提供一种玉龙杓兰共生真菌,该真菌的保藏名称为昆明巨丛子囊菌(Magnibotryascoma kunmingense) YAFEF024,保藏于中国典型培养物保藏中心,保藏地址为中国.武汉.武汉大学,保藏编号为CCTCC NO:M 2022613,保藏时间为2022年05月12日。
可选地,所述玉龙杓兰共生真菌包括如SEQ ID No.1与SEQ ID No.2所示的引物,所述玉龙杓兰共生真菌的ITS基因序列如SEQ ID No.3所示。
一方面,本申请还提供一种含有上述玉龙杓兰共生真菌在制备抗细菌药物中的应用。
另一方面,提供一种菌剂,包括上述的玉龙杓兰共生真菌的菌丝体粗提取物和/或细胞裂解上清液和/或细胞裂解沉淀。
可选地,所述菌丝体粗提取物的提取方法,包括以下步骤:
(1)将玉龙杓兰根部共生菌的菌丝体刮取规格为60mm培养皿中四分之一的菌丝放入2mL灭菌离心管中,再加入1mL无菌水,破碎2min,然后将破碎样品液取500µL分别接种到每个装有培养基的350mL的组培瓶中,置于26℃恒温培养箱中,培养15d;
所述培养基包括如下组分:山核桃渣8g/瓶与液体培养基15mL/瓶,所述液体培养基包括:硝酸钠6g/L、氯化钾0.52g/L、七水硫酸镁0.52g/L与磷酸二氢钾1.52g/L;
(2)将组培瓶培养的菌丝体置于60℃烘箱中干燥7h,去除培养基中的水分后,加入100mL甲醇超声震荡40min后静置48h,然后将菌液用漏斗进行分离,萃取溶液用旋转蒸发仪冷凝回流干燥,制得玉龙杓兰根部共生菌培养的菌丝体粗提物。
又一方面,本申请还提供一种玉龙杓兰共生真菌的分离方法,包括以下步骤:
(1)将采集的玉龙杓兰根部样品清洗干净后,用流动水冲洗24h,然后转入无菌工作台进行消毒处理。
(2)在无菌工作台中先用1L无菌水冲洗样品,放在培养皿中用接种刀切成小段放于50mL无菌离心管中;然后用体积分数为75%的乙醇浸泡1min期间不断震荡,倒掉乙醇后,用无菌水冲洗3次;无菌水冲洗后,倒入20mL配置好的体积分数为5%H2O2,浸泡3min,期间不断震荡,倒掉H2O2后,再用无菌水冲洗3次;将消毒后的样品用无菌滤纸吸干表面多余的水分,晾干备用。将玉龙杓兰根部切成长度与宽度均为0.5cm的组织块,在酒精灯火焰表面灭菌15s,把组织块等距离放在抗细菌培养基上培养,每个培养皿放置8个组织块,把培养皿做好标记,将培养皿倒置放于26℃培养箱中培养,培养8d,获取菌丝;所述抗细菌培养基包括如下组分:固体PDA培养基+氨苄青霉素50μg/mL+卡那霉素50μg/mL;
(3)将菌丝转接的到固体PDA培养基,培养8d后,对菌株进行分子鉴定,得到玉龙杓兰共生真菌。
可选地,在步骤(3)中,所述PDA固体培养基包括以下组分:马铃薯浸粉5g/L、酵母粉5g/L、葡萄糖20g/L、七水硫酸镁0.5g/L、磷酸二氢钾1g/L、维生素B10.1g/L与琼脂16g/L。
本发明的有益效果是:本发明通过分离筛选玉龙杓兰共生真菌,其菌丝体提取物对缓慢芽孢杆菌、无乳链球菌、短小芽孢杆菌、福氏志贺氏菌、枯草芽孢杆菌、藤黄微球菌等细菌具有较好的抗细菌活性。
本发明的菌株保藏信息如下:
菌株名称:昆明巨丛子囊菌(Magnibotryascoma kunmingense) YAFEF024;
保藏编号:CCTCC NO:M 2022613;
保藏单位名称:中国典型培养物保藏中心;
保藏地址:中国.武汉.武汉大学;
保藏时间:2022年05月12日。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为本发明玉龙杓兰共生真菌的菌丝生长情况图;
图2为本发明玉龙杓兰共生真菌的抗菌活性检测结果图,a表示显著性差异分析结果,都是a表示数据可信度高;
图3为本发明玉龙杓兰共生真菌的荧光显微镜下菌丝体图;
图4为本发明玉龙杓兰共生真菌对缓慢芽孢杆菌的抑制作用图,a、b、d表示对照组,c表示实验组;
图5为本发明玉龙杓兰共生真菌对无乳链球菌的抑制作用图,a、b、d表示对照组,c表示实验组;
图6为本发明玉龙杓兰共生真菌对短小芽孢杆菌的抑制作用图,a、b、d表示对照组,c表示实验组;
图7为本发明玉龙杓兰共生真菌对福氏志贺氏菌的抑制作用图,a、b、d表示对照组,c表示实验组;
图8为本发明玉龙杓兰共生真菌对枯草芽孢杆菌的抑制作用图,a、b、d表示对照组,c表示实验组;
图9为本发明玉龙杓兰共生真菌对藤黄微球菌的抑制作用图,a、b、d表示对照组,c表示实验组;
图10本发明玉龙杓兰共生真菌细胞的超声裂解上清和细胞的超声裂解沉淀样品对缓慢芽孢杆菌的抑制作用图,a表示对照组,b、c表示实验组;
图11本发明基于ITS构建了玉龙杓兰共生真菌的系统发育树显示图。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚,下面将对本发明的技术方案进行详细的描述。显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动的前提下所得到的所有其它实施方式,都属于本发明所保护的范围。
本发明的菌株保藏信息如下:
菌株名称:昆明巨丛子囊菌(Magnibotryascoma kunmingense) YAFEF024;
保藏编号:CCTCC NO:M 2022613;
保藏单位名称:中国典型培养物保藏中心;
保藏地址:中国.武汉.武汉大学;
保藏时间:2022年05月12日。
实施例
玉龙杓兰共生真菌YAFEF024菌株的分离和鉴定
1、玉龙杓兰共生菌的分离
将采集的玉龙杓兰根部样品用自来水冲洗24h,清除样品表面的真菌或者其他微生物,然后转入无菌工作台进行消毒处理,先用1L无菌水冲洗样品,放在培养皿中用接种刀切成小段放于50mL无菌离心管中;然后用体积分数为75%的乙醇浸泡1min(期间不断震荡),倒掉乙醇后,用无菌水冲洗3次;无菌水冲洗后,倒入20mL配置好的体积分数为5%H2O2,浸泡3min(期间不断震荡),倒掉H2O2后,再用无菌水冲洗3次;将消毒后的样品用无菌滤纸吸干表面多余的水分,晾干备用。
然后用灭菌接种刀将玉龙杓兰根部样品切成长宽0.5cm大小的组织块,接种在PDAKAS抗细菌培养基上(固体PDA培养基+氨苄青霉素50μg/mL+卡那霉素50μg/mL)培养,将培养皿倒置放于26℃培养箱中培养,期间不定期观察。将长出的菌丝转接到固体PDA培养基(马铃薯浸粉5g/L、酵母粉5g/L、葡萄糖20g/L、七水硫酸镁0.5g/L、磷酸二氢钾1g/L、维生素B10.1g/L与琼脂16g/L)培养8d后,对菌株进行分子鉴定,得到玉龙杓兰根部共生真菌。
2、玉龙杓兰共生真菌的鉴定
(1)形态的鉴定
如图1所示,玉龙杓兰共生真菌在固体PDA培养基上生长良好,菌丝颜色呈现白色,菌落表面平整呈放射状向周围生长,且菌层较厚,菌落呈不规则形状。
(2)DNA提取
①实验前先将CTAB(十六烷基三甲基溴化铵)放在65℃水浴锅中加热30min;②取50mg烘干的玉龙杓兰共生真菌菌丝体于2mL的离心管中,加入3颗小钢珠,将该离心管放入液氮中浸泡6min,并立即用破碎机破碎1.5min打碎,加入1mL预热的CTAB溶液,用移液枪吸打混匀后全部移入装有200μLPVP(聚乙烯基吡咯烷酮)的离心管中,在通风橱中悬空添加20μLβ-巯基乙醇,震荡15s以充分研磨,然后放到65oC水浴锅水浴1.5h,每10min上下翻转5-6次,水浴结束后12000r/min,4℃离心10min;③取1mL上清液到新的离心管中,加入DNA酚试剂、氯仿-异戊醇混合液各500μL,上下翻转10min,离心(4℃·12000r/min)10min(③步骤重复两次);④取上清液900μL放入一个新的离心管中,加入50μL 3mol醋酸钠溶液和900μL95%无水冰乙醇(-20℃),摇匀放入-20℃冰箱沉淀3h;⑤沉淀完离心(4℃·12000r/min)10min,弃上清液,加入500μL75%酒精上下翻转2-3次,静置3min,弃上清液;⑥加入500μL95%酒精上下翻转2-3次,静置3min,常温离心(13000r)3min,弃乙醇,放置干;⑦加入40μL洗脱缓冲液EB,常温离心(13000/min)1.5min,得到玉龙杓兰共生真菌基因组DNA。
(3)ITS分析鉴定:用真菌SEQ ID No.1所示的引物ITS1(5’-CTTGGTCATTTAGAGGAAGTAA-3’)和SEQ ID No.2所示的引物ITS4(5’-TCCTCCGCTTATTGATATGC-3’)扩增真菌rDNA的间隔序列(含ITS1区、5.8S区、ITS4区)序列,所得得ITS测序序列如SEQ ID No.3所示:
ACGGCATCTACCTGATTCGAGGTCAACAAGTGAAATAGGTTCGCTTCTGGGAGCCAGGCCGCGGGCGAGGAGCGCAATTCTGCTGCACTCCAGGCCGTGGCACCAGCCGCCAATCGCTTTGAGGCGAGTCCGCGCCCGAAGGCGGGACAGACGCCCAACACCAAGCAGTGCTTGAGGGTGTAAATGACGCTCGAACAGGCATGCCCCACGGAATACCGAGGGGCGCAATGTGCGTTCAAAGATTCGATGATTCACGGAATTCTGCAATTCACACTACTTATCGCATTTCGCTGCGTTCTTCATCGATGCCAGAGCCAAGAGATCCATTGTTGAAAGTTGTGATTAATTTTTTTATGATACTGTTACTTCTCATTACAAAAGGTTTTTGAGGGTCCCCAGTGGCGGGCAAGCCCGCCGAGGAAACGAAATGGTGCTCAAAAGGCAGGGTGGCCCGGCGATATGCTCGGGGGGTTGCTGGCGCACCTTACCCCACCTAATTTTCGCTAGGTGGGGACTCGGCTGCCAGTCCCTATGCGTAATGATCCTTCCGCAGGTTCACCTACGGAAACCTTGTTACGACTTTTACTTCCTCAA。
(4)构建发育树
基于ITS利用MEGA软件构建如图11所示的玉龙杓兰共生真菌的系统发育树,综合菌株形态学鉴定和分子生物学鉴定结果,该真菌与昆明巨丛子囊菌(Magnibotryascomakunmingense)亲缘关系最近,所以将YAFEF024菌株鉴定为昆明巨丛子囊菌(Magnibotryascoma kunmingense)。
玉龙杓兰共生菌YAFEF024活性物质的提取
1、固体发酵培养
将已长成型的玉龙杓兰根部共生菌YAFEF024菌丝体刮取60mm培养皿中四分之一的菌丝放入2mL灭菌离心管中,再加入1mL无菌水,破碎2min,然后将破碎样品液取500µL分别接种到每个装有CCMM培养基的组培瓶中,置于恒温培养箱26℃,培养15d。
2、培养物菌丝体提取
将组培瓶培养的菌丝体置于60℃烘箱中干燥7h,去除培养基中的水分后,加入100mL甲醇超声震荡40min后静置48h,然后将菌液用漏斗进行分离,萃取溶液用旋转蒸发仪冷凝回流干燥,制得玉龙杓兰根部共生菌YAFEF024培养的菌丝体粗提物。
试验例1
玉龙杓兰共生菌YAFEF024培养物的菌丝体粗提物的抗菌活性检测
1、致病菌的活化
将购于广东省细菌耐药性监测和质量控制中心的缓慢芽孢杆菌、无乳链球菌、短小芽孢杆菌、福氏志贺氏菌、枯草芽孢杆菌、藤黄微球菌等细菌,分别取5μL加入2mL离心管中,再加入750μL的LB液体培养基(胰蛋白胨10g/L+氯化钠10g/L+酵母提取物5g/L),然后将离心管放入37℃,转速为180 r·min-1恒温摇床中培养12h,得到致病菌菌液,置于4℃冰箱备用,取出活化好的细菌进行稀释1/10倍。
2、对照所用抗生素及配制
氨苄青霉素(Ampicillin)(批号:0339,纯度:95%),购于昆明硕阳科技有限公司。精密称取氨苄青霉素50mg,分别置于离心管中,加入1mL DMSO溶液配置成50mg/ml的抗生素DMSO溶液,备用。
3、滤纸片法检测抗菌活性
取0.01g玉龙杓兰共生菌YAFEF024菌丝体粗提物于2mL离心管,加入500μL DMSO(二甲基亚砜)溶液,稀释到50mg/mL。将活化得到的致病菌液分别均匀涂抹在LB固体培养基(胰蛋白胨10g/L+氯化钠10g/L+琼脂15g/L+酵母提取物5g/L)上,晾干后,将吸取了已溶解的玉龙杓兰共生菌YAFEF024菌丝体粗提物的5mm的滤纸圆片放在培养基的对应标记处,并用5mm滤纸圆片蘸取纯培养基(操作步骤和实验组一样,只是做空白对照)、DMSO溶液和氨苄青霉素作为对照,将培养基正放置于37℃的恒温培养箱中培养12h,观察是否出现抑菌圈并用十字交叉法量出抑菌圈的直径大小。玉龙杓兰共生菌YAFEF024培养物的菌丝体粗提物(50mg/mL)的抗细菌活性结果如图2所示。图2是统计该菌对6种细菌的抑菌效果,用柱形图展示,缓慢芽孢杆菌、无乳链球菌、短小芽孢杆菌、福氏志贺氏菌、枯草芽孢杆菌、藤黄微球菌,抑菌直径分别为1.4cm、1.4cm、1.3cm、1.3cm、1.2cm与1.4cm。
本发明筛选得到一种玉龙杓兰共生真菌YAFEF024(昆明巨丛子囊菌Magnibotryascoma kunmingense),其菌株培养物的菌丝体粗提物对缓慢芽孢杆菌、无乳链球菌、短小芽孢杆菌、福氏志贺氏菌、枯草芽孢杆菌、藤黄微球菌等细菌具有较好的抗细菌活性。
玉龙杓兰共生真菌YAFEF024细胞的超声裂解上清和细胞的超声裂解沉淀操作步骤及抗菌活性检测
将已长成型的玉龙杓兰共生真菌YAFEF024菌丝体刮取3个60mm培养皿中的菌丝放入离心管中,再用真空冷冻干燥剂冷冻,后加入100mL甲醇浸提,超声震荡40min后静置48h,分别获取上清液和沉淀液,再用旋转蒸发仪冷凝回流干燥,制得玉龙杓兰共生真菌YAFEF024细胞的超声裂解上清及细胞的超声裂解沉淀样品,将这些样品用于做抗菌活性检测。其对照是将已长成型的玉龙杓兰共生真菌YAFEF024菌丝体刮取60mm培养皿中四分之一的菌丝放入2mL灭菌离心管中,再加入1mL无菌水,破碎2min,然后将破碎样品液取500µL接种到装有CCMM培养基的组培瓶中,置于恒温培养箱26℃,培养10d。将组培瓶培养的菌丝体置于60℃烘箱中干燥7h,去除培养基中的水分后,加入100mL甲醇浸提,超声震荡40min后静置48h,然后将菌液用漏斗进行分离,萃取溶液用旋转蒸发仪冷凝回流干燥,制得玉龙杓兰共生真菌YAFEF024培养的菌丝体粗提物,后做抗菌活性检测。
除了通过培养菌株进行菌丝体提取,其他培养或者加工方法,如玉龙杓兰共生真菌YAFEF024细胞的超声裂解上清;玉龙杓兰共生真菌YAFEF024细胞的超声裂解沉淀,基于真菌YAFEF024的抗菌效果下,其加工物或者培养物均具备相应的抗菌活性,使用本菌制备相关抗菌活性药物、菌剂等,均在本发明的保护范围内。
综上,图4-图9,a代表纯培养基(CCMM培养基)就是未接种该菌空白培养基的粗提物;b代表DMSO溶液;c代表昆明巨丛子囊菌Magnibotryascoma kunmingense YAFEF024在CCMM培养基培养获得的粗提物;d代表氨苄青霉素溶液。在试验例1中已经提到过。得出的结论是确实是该菌对细菌有抑菌效果,a,b排除了培养基、DMSO有抑菌效果的可能性,d说明常用广谱抗生素(氨苄青霉素)对本专利中所使用细菌无抗菌效果,证明昆明巨丛子囊菌Magnibotryascoma kunmingense YAFEF024抗菌活性开发的意义。
如图10所示,图中a代表昆明巨丛子囊菌Magnibotryascoma kunmingenseYAFEF024在CCMM培养基培养获得的粗提物;b.代表昆明巨丛子囊菌Magnibotryascomakunmingense YAFEF024细胞的超声裂解上清样品;c. 昆明巨丛子囊菌Magnibotryascomakunmingense YAFEF024细胞的超声裂解沉淀样品;得出的结论是细胞的超声裂解有助于该菌中抑菌活性成分的释放。
以上所述,仅为本发明的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明记载的技术范围内,可轻易想到变化或替换,都应涵盖在本发明的保护范围之内。因此,本发明的保护范围应以所述权利要求的保护范围为准。
Claims (7)
1.一种玉龙杓兰共生真菌,其特征在于,该真菌的保藏名称为昆明巨丛子囊菌(Magnibotryascoma kunmingense)YAFEF024,保藏于中国典型培养物保藏中心,保藏地址为中国.武汉.武汉大学,保藏编号为CCTCC NO:M 2022613,保藏时间为2022年05月12日。
2.根据权利要求1所述的一种玉龙杓兰共生真菌,其特征在于,所述玉龙杓兰共生真菌包括如SEQ ID No.1与SEQ ID No.2所示的引物,所述玉龙杓兰共生真菌的ITS基因序列如SEQ ID No.3所示。
3.一种如权利要求1或2所述的玉龙杓兰共生真菌在制备抗细菌药物中的应用。
4.一种菌剂,其特征在于,包括权利要求1或2所述的玉龙杓兰共生真菌的菌丝体粗提取物和/或细胞裂解上清液和/或细胞裂解沉淀。
5.根据权利要求4所述的菌剂,其特征在于,所述菌丝体粗提取物的提取方法,包括以下步骤:
(1)将玉龙杓兰根部共生菌的菌丝体刮取规格为60mm培养皿中四分之一的菌丝放入2mL灭菌离心管中,再加入1mL无菌水,破碎2min,然后将破碎样品液取500µL分别接种到每个装有培养基的350mL的组培瓶中,置于26℃恒温培养箱中,培养15d;
所述培养基包括如下组分:山核桃渣8g/瓶与液体培养基15mL/瓶,所述液体培养基包括:硝酸钠6g/L、氯化钾0.52g/L、七水硫酸镁0.52g/L与磷酸二氢钾1.52g/L;
(2)将组培瓶培养的菌丝体置于60℃烘箱中干燥7h,去除培养基中的水分后,加入100mL甲醇超声震荡40min后静置48h,然后将菌液用漏斗进行分离,萃取溶液用旋转蒸发仪冷凝回流干燥,制得玉龙杓兰根部共生菌培养的菌丝体粗提物。
6.一种玉龙杓兰共生真菌的分离方法,其特征在于,包括以下步骤:
(1)将采集的玉龙杓兰根部样品清洗干净后,用流动水冲洗24h,然后转入无菌工作台进行消毒处理;
(2)在无菌工作台中先用1L无菌水冲洗样品,放在培养皿中用接种刀切成小段放于50mL无菌离心管中;然后用体积分数为75%的乙醇浸泡1min期间不断震荡,倒掉乙醇后,用无菌水冲洗3次;无菌水冲洗后,倒入20mL配置好的体积分数为5%H2O2,浸泡3min,期间不断震荡,倒掉H2O2后,再用无菌水冲洗3次;将消毒后的样品用无菌滤纸吸干表面多余的水分,晾干备用,将玉龙杓兰根部切成长度与宽度均为0.5cm的组织块,在酒精灯火焰表面灭菌15s,把组织块等距离放在抗细菌培养基上培养,每个培养皿放置8个组织块,把培养皿做好标记,将培养皿倒置放于26℃培养箱中培养,培养8d,获取菌丝;
所述抗细菌培养基包括如下组分:固体PDA培养基+氨苄青霉素50μg/mL+卡那霉素50μg/mL;
(3)将菌丝转接的到固体PDA培养基,培养8d后,对菌株进行分子鉴定,得到玉龙杓兰共生真菌。
7.根据权利要求6所述的分离方法,其特征在于,在步骤(3)中,所述固体PDA培养基包括以下组分:马铃薯浸粉5g/L、酵母粉5g/L、葡萄糖20g/L、七水硫酸镁0.5g/L、磷酸二氢钾1g/L、维生素B10.1g/L与琼脂16g/L。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310611837.9A CN116426394B (zh) | 2023-05-29 | 2023-05-29 | 一种玉龙杓兰共生真菌及应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310611837.9A CN116426394B (zh) | 2023-05-29 | 2023-05-29 | 一种玉龙杓兰共生真菌及应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN116426394A true CN116426394A (zh) | 2023-07-14 |
CN116426394B CN116426394B (zh) | 2023-08-29 |
Family
ID=87083512
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310611837.9A Active CN116426394B (zh) | 2023-05-29 | 2023-05-29 | 一种玉龙杓兰共生真菌及应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116426394B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117737138A (zh) * | 2024-02-07 | 2024-03-22 | 云南省林业和草原科学院 | 一种诱导高黎贡牛樟芝产生抑制植物病害细菌化合物的培养方法 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB8400015D0 (en) * | 1984-01-03 | 1984-02-08 | Clements M A | Germination growth of orchid seeds |
CN102492628A (zh) * | 2011-12-06 | 2012-06-13 | 北京市植物园 | 一种促进大花杓兰种子共生萌发的真菌及其应用 |
CN106557624A (zh) * | 2016-11-14 | 2017-04-05 | 中国科学院华南植物园 | 一种兰科植物濒危状态评估的方法 |
CN115568395A (zh) * | 2022-11-21 | 2023-01-06 | 中国科学院昆明植物研究所 | 一种提高玉龙杓兰结实率及获取饱满种子的方法 |
-
2023
- 2023-05-29 CN CN202310611837.9A patent/CN116426394B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB8400015D0 (en) * | 1984-01-03 | 1984-02-08 | Clements M A | Germination growth of orchid seeds |
CN102492628A (zh) * | 2011-12-06 | 2012-06-13 | 北京市植物园 | 一种促进大花杓兰种子共生萌发的真菌及其应用 |
CN106557624A (zh) * | 2016-11-14 | 2017-04-05 | 中国科学院华南植物园 | 一种兰科植物濒危状态评估的方法 |
CN115568395A (zh) * | 2022-11-21 | 2023-01-06 | 中国科学院昆明植物研究所 | 一种提高玉龙杓兰结实率及获取饱满种子的方法 |
Non-Patent Citations (4)
Title |
---|
LIU 等: "In vitro conservation of Paphiopedilum wenshanense at Kunming Botanical Garden, China", ORYX, vol. 56, no. 6, pages 811 - 812 * |
MORTIMER 等: "Morpho-Phylo Taxonomy of Novel Dothideomycetous Fungi Associated With Dead Woody Twigs in Yunnan Province, China", FRONTIERS IN MICROBIOLOGY, vol. 12, pages 654683 * |
安曼云 等: "云南杓兰菌根真菌组成及共生关系研究", 广西植物, vol. 37, no. 6, pages 763 - 767 * |
徐玲玲 等: "三种杓兰根相关真菌多样性和生态功能", 微生物学通报, vol. 46, no. 9, pages 2134 - 2145 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117737138A (zh) * | 2024-02-07 | 2024-03-22 | 云南省林业和草原科学院 | 一种诱导高黎贡牛樟芝产生抑制植物病害细菌化合物的培养方法 |
CN117737138B (zh) * | 2024-02-07 | 2024-05-28 | 云南省林业和草原科学院 | 一种诱导高黎贡牛樟芝产生抑制植物病害细菌化合物的培养方法 |
Also Published As
Publication number | Publication date |
---|---|
CN116426394B (zh) | 2023-08-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11459593B2 (en) | Dendrobium officinale endophytic fungus strain and extracellular polysaccharide produced thereby, and extraction method and application of extracellular polysaccharide | |
CN107828665B (zh) | 一种竹叶兰内生真菌的分离纯化方法及其应用 | |
CN115044505B (zh) | 一株贝莱斯芽孢杆菌产抗菌脂肽及其在化妆品和食品的应用 | |
CN116426394B (zh) | 一种玉龙杓兰共生真菌及应用 | |
CN112175887B (zh) | 一种油短波单胞菌菌株及其应用 | |
CN112812971B (zh) | 一种青头菌共生真菌m2-1及其菌剂和发酵液提取物 | |
CN108865895A (zh) | 蝙蝠蛾拟青霉zjb18001及其应用 | |
CN113881571A (zh) | 一种快速分离、定量检测香蕉枯萎病菌的方法及应用 | |
CN116355763B (zh) | 一种油麦吊云杉共生真菌及应用 | |
CN118126845A (zh) | 一株长松萝内生真菌yafef048菌株及其分离方法和应用 | |
CN112574894B (zh) | 一种伊氏杀线虫真菌及其应用 | |
CN108004154B (zh) | 腥掷孢酵母菌17wy1、其微生物制剂及其在小麦白粉病防治中的应用 | |
CN114958616A (zh) | 一种香樟共生真菌yafef008及其分离方法 | |
CN116083243B (zh) | 一种牛樟芝与节菱孢霉复合生防菌剂的制备方法 | |
Kaushik et al. | Identification, optimization of culture conditions, and bioactive potential of Chinese caterpillar mushroom Ophiocordyceps sinensis (Ascomycetes) mycelium isolated from fruiting body | |
CN115197853A (zh) | 内生菌Epicoccum thailandicumLF-28菌株及其应用 | |
CN116262902A (zh) | 一种持续诱导沉香积累的真菌及其应用 | |
CN111004727B (zh) | 一株在高盐环境下增加木麻黄生物量的内生真菌z1 | |
CN116064242B (zh) | 兰淡领瓶霉及其分离方法和应用 | |
CN116064241B (zh) | 盾壳霉yafef037菌株及其分离方法和应用 | |
CN111500494A (zh) | 一株具有高酶活的蒂莫内马赛菌及其筛选方法和应用 | |
CN114214205B (zh) | 牛樟芝共生真菌AcEF007及其分离方法 | |
CN115786130B (zh) | 丽江杓兰共生真菌yafef040及应用 | |
CN116004392B (zh) | 一种牛樟共生真菌yafef009及其分离方法 | |
CN118109317B (zh) | 一株Chloridium gonytrichii DZW4及在制备抑制病原微生物制剂中的应用 |
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 |