CN107523506B - 一种菌根真菌及其在抗肿瘤中的应用 - Google Patents
一种菌根真菌及其在抗肿瘤中的应用 Download PDFInfo
- Publication number
- CN107523506B CN107523506B CN201710771269.3A CN201710771269A CN107523506B CN 107523506 B CN107523506 B CN 107523506B CN 201710771269 A CN201710771269 A CN 201710771269A CN 107523506 B CN107523506 B CN 107523506B
- Authority
- CN
- China
- Prior art keywords
- mycorrhizal fungi
- fungi
- pleat
- purple
- present
- 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.)
- Active
Links
- 241000233866 Fungi Species 0.000 title claims abstract description 35
- 230000000259 anti-tumor effect Effects 0.000 title abstract description 8
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 4
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 4
- 241000222433 Tricholomataceae Species 0.000 claims description 10
- 241000222485 Agaricales Species 0.000 claims description 6
- 241000121219 Tricholoma Species 0.000 claims description 5
- 241000128201 Tricholosporum Species 0.000 claims description 3
- 235000001674 Agaricus brunnescens Nutrition 0.000 abstract description 15
- 238000002955 isolation Methods 0.000 abstract description 6
- 210000004881 tumor cell Anatomy 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 240000008042 Zea mays Species 0.000 description 11
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 11
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 11
- 235000005822 corn Nutrition 0.000 description 11
- 239000008367 deionised water Substances 0.000 description 11
- 229910021641 deionized water Inorganic materials 0.000 description 11
- 235000013312 flour Nutrition 0.000 description 11
- 239000000284 extract Substances 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 9
- 238000000855 fermentation Methods 0.000 description 8
- 230000004151 fermentation Effects 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 239000012138 yeast extract Substances 0.000 description 8
- 229920001817 Agar Polymers 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 7
- 239000008272 agar Substances 0.000 description 7
- 229940041514 candida albicans extract Drugs 0.000 description 7
- 239000000843 powder Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 230000001954 sterilising effect Effects 0.000 description 7
- 239000007788 liquid Substances 0.000 description 6
- 241000221198 Basidiomycota Species 0.000 description 5
- -1 Ucleosides Chemical class 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 238000004659 sterilization and disinfection Methods 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 241000139962 Tricholosporum sp. Species 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 244000178870 Lavandula angustifolia Species 0.000 description 3
- 235000010663 Lavandula angustifolia Nutrition 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 239000003864 humus Substances 0.000 description 3
- 239000001102 lavandula vera Substances 0.000 description 3
- 235000018219 lavender Nutrition 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 241000009794 Agaricomycetes Species 0.000 description 2
- 244000251953 Agaricus brunnescens Species 0.000 description 2
- 240000008397 Ganoderma lucidum Species 0.000 description 2
- 235000001637 Ganoderma lucidum Nutrition 0.000 description 2
- 241000414067 Inonotus obliquus Species 0.000 description 2
- 244000197580 Poria cocos Species 0.000 description 2
- 235000008599 Poria cocos Nutrition 0.000 description 2
- 241000121220 Tricholoma matsutake Species 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 231100000225 lethality Toxicity 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 241000221377 Auricularia Species 0.000 description 1
- 241001489099 Cantharellus Species 0.000 description 1
- 241001496872 Clavaria Species 0.000 description 1
- 235000002469 Clitocybe nuda Nutrition 0.000 description 1
- 241001236122 Coprinopsis atramentaria Species 0.000 description 1
- 241000222511 Coprinus Species 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 240000006499 Flammulina velutipes Species 0.000 description 1
- 235000016640 Flammulina velutipes Nutrition 0.000 description 1
- 241000222336 Ganoderma Species 0.000 description 1
- 241000222684 Grifola Species 0.000 description 1
- 240000001080 Grifola frondosa Species 0.000 description 1
- 235000007710 Grifola frondosa Nutrition 0.000 description 1
- 241000577951 Hydnum Species 0.000 description 1
- 108091023242 Internal transcribed spacer Proteins 0.000 description 1
- 208000034693 Laceration Diseases 0.000 description 1
- 241000446313 Lamella Species 0.000 description 1
- 240000000599 Lentinula edodes Species 0.000 description 1
- 235000001715 Lentinula edodes Nutrition 0.000 description 1
- 244000290281 Lepista nuda Species 0.000 description 1
- 244000028178 Marasmius oreades Species 0.000 description 1
- 235000017233 Marasmius oreades Nutrition 0.000 description 1
- 244000168667 Pholiota nameko Species 0.000 description 1
- 235000014528 Pholiota nameko Nutrition 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 241001506047 Tremella Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
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
-
- 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
-
- 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
- A61K36/07—Basidiomycota, e.g. Cryptococcus
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medical Informatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
本发明公开一种菌根真菌，该菌根真菌从紫褶口蘑采用组织分离法得到的纯培养菌种，保藏于中国典型培养物保藏中心，保藏编号为：CCTCC NO:M 2017185，保藏菌株号为HMGIM‑D160241。本发明的菌根真菌，能够有效抑制肿瘤细胞，可用于制备抗肿瘤药物等，具有较好的抗肿瘤效果。
Description
技术领域
本发明涉及一种真菌及其应用，尤其是一种从紫褶口蘑进行纯培养得到的菌根真菌及其在抗肿瘤中的应用。
背景技术
恶性肿瘤是一类严重危害人类健康和生命的疾病，在所有死亡原因中已经超过心脑血管疾病、成为排名第一的死亡原因。随着自然生态的失衡和环境污染的加剧，恶性肿瘤的发病率和死亡率在世界范围内呈现着上升趋势。
大型真菌代谢产物含有丰富的活性物质，如具有抗肿瘤功效的真菌多糖、甾醇类、核苷类、萜类化合物、真菌蛋白等，现有报道具有抗肿瘤活性的大型真菌有香菇(Lentinusedodes)、灵芝(Ganoderma spp)、茯苓(Poria cocos)、金针菇(Flammulina velutipes)、黑木耳(Auricularia auricular)、鸡油菌(Cantharellus spp)、珊瑚菌(Clavaria spp)、齿菌(Hydnum spp)、香蘑(Lepista nuda)、松茸(Tricholoma matsutake)、双孢蘑菇(Agaricus bisporus)、墨汁鬼伞(Coprinus atramentarius)、滑菇(Pholiota nameko)、栓菌(Tremella spp)、灰树花(Grifolafrondosa)、桦褐孔菌(Inonotus obliquus)等多孔菌，这些大型真菌子实体或发酵液的提取物所具有的活性成分，已经开始被开发，特别是香菇多糖、灵芝多糖、灵芝三萜等功效显著的化合物已经被明晰，因而从大型真菌中继续寻找有新的作用机制和特殊化学结构的细胞药物具有重大潜力，成为癌症防治研究的一个新的热点。
发明内容
本发明的目的在于结合上述背景技术寻找一种新的具有抗肿瘤效果的菌根真菌。
为实现上述目的，本发明采取的技术方案为：一种菌根真菌，所述菌根真菌保藏于中国典型培养物保藏中心，保藏编号为：CCTCC M 2017185，保藏菌株号为HMGIM-D160241，拉丁文分类命名为：Tricholosporum sp.。
本发明的菌根真菌从紫褶口蘑采用组织分离法得到的纯培养菌种，所述紫褶口蘑来自真菌界(Fungi)担子菌门(Basidiomycota)伞菌纲(Agaricomycetes)伞菌目(Agaricales)口蘑科(Tricholomataceae)紫褶口蘑属。紫褶口蘑(Tricholosporum sp.)为口蘑科(Tricholomataceae)、紫褶口蘑属(Tricholosporum)的一个新物种，采自辽宁阜新三塔沟风景区内玉米地旁的腐殖层，采用组织分离法得到纯培养菌种，并于2017年4月10日保藏于中国典型培养物保藏中心(地址：中国，武汉，武汉大学，邮编430072)，保藏编号：CCTCC M 2017185，保藏菌株号：HMGIM-D160241，拉丁文分类命名为：Tricholosporum sp.，子实体及菌种的rDNA-ITS序列已提交至NCBI，登记号分别为：MF538720，MF538721。
另外，本发明提供了如上所述菌根真菌的制备方法，所述菌根真菌为紫褶口蘑采用组织分离法得到的纯培养菌种。
作为本发明所述菌根真菌的制备方法的优选实施方式，采用组织分离法进行培养时，采用的培养基包含：玉米粉水提物、麦芽提取物、酵母提取物、琼脂粉和去离子水。采用组织分离方法在MMN、PDA等通用培养基上接种组织块无法萌发获得纯培养，而在改良的复合培养基上生长情况较好，其配方包含：玉米粉水提物、麦芽提取物、酵母提取物、琼脂粉和去离子水。
作为本发明所述菌根真菌的制备方法的优选实施方式，所述培养基中，各组分的质量浓度为：玉米粉水提物50g/L、麦芽提取物20g/L、酵母提取物3g/L、琼脂粉20g/L。作为本发明所述菌根真菌的制备方法的更优选实施方式，所述培养基中，含玉米粉水提物50g、麦芽提取物20.0g、酵母提取物3.0g、琼脂粉20.0g、去离子水1000ml，121℃高压灭菌15分钟。在该培养基上菌丝呈现淡紫色、有气生菌丝，基内菌丝较弱，且生长一段时间(10d)后形成拧结状，质地绒状。
作为本发明所述菌根真菌的制备方法的优选实施方式，所述培养基采用如下方法配置而成：称取玉米粉，溶解至一部分去离子水，煮沸，15min后使用3层纱布过滤，保存滤液，然后称取麦芽提取物、酵母提取物，溶解至玉米粉提取液，继续添加一部分去离子水，加热并完全溶解后，称取琼脂粉并不断搅拌至完全溶解，然后添加剩余去离子水定容，分装至三角瓶或试管中，加胶塞密封，121℃高压灭菌20分钟，灭菌完成后待温度降至60℃后取出，洁净条件下倒平板或摆斜面，制成平板或斜面试管培养基。
最后，本发明还提供了所述菌根真菌在制备抗肿瘤的药物中的用途。本发明中所述菌根真菌的菌丝体发酵液，能够有效抑制肿瘤细胞。
附图说明
图1为本发明菌根真菌培养10天的生长状况图。
图2为本发明菌根真菌的子实体图。
图3为本发明菌根真菌子实体的担孢子扫描电镜图。
具体实施方式
为更好的说明本发明的目的、技术方案和有益效果，下面将结合附图和具体实施例对本发明作进一步说明。
实施例1
本发明菌根真菌的一种实施例，本实施例所述菌根真菌从紫褶口蘑采用组织分离法得到的纯培养菌种，所述紫褶口蘑来自真菌界(Fungi)担子菌门(Basidiomycota)伞菌纲(Agaricomycetes)伞菌目(Agaricales)口蘑科(Tricholomataceae)紫褶口蘑属，是口蘑科(Tricholomataceae)、紫褶口蘑属(Tricholosporum)的一个新物种，采自辽宁阜新三塔沟风景区内玉米地旁的腐殖层。
该新种是通过野外采集得到，依据形态学和分子生物学鉴定，菌种是分离自新鲜的子实体，经过多次纯化培养获得，并使用ITS序列验证与子实体一致，纯化培养基配置方法如下：称取玉米粉50g，溶解至500ml去离子水，煮沸，15min后使用3层纱布过滤，保存滤液，然后称取麦芽提取物20.0g，酵母提取物3.0g，溶解至玉米粉提取液，继续添加400ml去离子水，加热并完全溶解后，称取琼脂粉20.0g并不断搅拌至完全溶解，然后添加去离子水定容至1000ml，分装至250ml三角瓶或试管中，加胶塞密封，121℃高压灭菌20分钟，灭菌完成后待温度降至60℃后取出，洁净条件下倒平板或摆斜面，制成平板或斜面试管培养基。
子实体及菌丝体经试剂盒提取DNA，并以ITS1/ITS4引物扩增和测序，子实体和菌丝体共同的rDNA-ITS序列如下：
GGAAGGATCATTATTGAATGAACTTGGTCCAAGTTGTTGCTGGCTCCTTGGAGCATATATGTGTGCACGCTTGGTACTCATTCTATTTACCCACCTGTGCACTCTTTTGTAGAGCCCTGAGATATTTAGTTATTCAGGTGTTTCCTGAGTTGAAGGTTTGCTGTGCGAAAGTCAGCTTTCCTTTATTTTTTATCTCTAGGGATTCTATGTATTCTTATATACTCCTAATGAATGTAACAGAATGTTATGTTGGTTTTAAAAAAACACAATATACAACTTTCAACAATGGATCTCTTGGCTCTCGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCATCTTGCGCCCCTTGGTATTCCGAGGAGCATGCCTGTTTGAGTGTCATTAAATTTCTCAATCCTTCCTTTCGTTCTCAATGAATGAAAGAGGTTGGCATTGGAATGTGGAAGCTGCAGGCTTTTTATTATTATAATAAAGTCAGCTCTTCTAAAAAGCATTAGCTAAATGTTTTGAGAATTAAACTAGCATTGGTGTGATAATTATCTACGCCATTTGTTAAATCTCTTCATAGTTTAGCTTCTAATATAACATTGGTCTTTGGGACCTTGTTTTGATAATTTGACCTCAAATCAGGTAGGACTACCCGCTGAACTTAAGCATATCAA
采用组织分离法得到的纯培养菌种即为本实施例的菌根真菌，已于2017年4月10日保藏于中国典型培养物保藏中心(地址：中国，武汉，武汉大学，邮编430072)，保藏编号：CCTCC M 2017185，保藏菌株号：HMGIM-D160241，拉丁文分类命名为：Tricholosporum sp.，子实体及菌种的rDNA-ITS序列已提交至NCBI，登记号分别为：MF538720，MF538721。
采用组织分离方法在MMN、PDA等通用培养基上接种组织块无法萌发获得纯培养，而在改良的复合培养基上生长情况较好，其配方为：玉米粉50g水提物，麦芽提取物20.0g，酵母提取物3.0g，琼脂粉20.0g，去离子水1000ml，121℃高压灭菌15分钟。在该培养基上菌丝呈现淡紫色、有气生菌丝，基内菌丝较弱，且生长一段时间(10d)后形成拧结状，质地绒状，如附图1所示。
子实体群生于阔叶林腐殖层上，常形成蘑菇圈，菌盖平展，淡黄色至橙色，带部分紫色，中部略凸起，菌盖边缘撕裂状，有绒毛状附属物，并呈波浪状，干燥时部分内卷，菌褶淡紫色，直生至弯生，不等长，褶缘近似锯齿状，菌柄圆柱形，下部稍粗，有绒毛状附属物，中下部颜色稍深，颜色近菌盖，上部白色，菌柄实心，纤维质，菌肉白色，如附图2所示。担孢子十字型，6.1～6.5×5.5～5.9μm，担孢子扫描电镜图如附图3所示。此种的主要特点是担孢子呈十字型。
实施例2本发明菌根真菌抗肿瘤的效果试验
菌丝体发酵液的制备方法：首先准备发酵培养基：称取玉米粉50g，溶解至500ml去离子水，煮沸，15min后使用3层纱布过滤，保存滤液，然后称取麦芽提取物20.0g，酵母提取物3.0g，溶解至玉米粉提取液，继续添加400ml去离子水，加热并完全溶解后，添加去离子水定容至1000ml，分装至250ml三角瓶中，每个三角瓶定量100ml，加胶塞密封，121℃高压灭菌20分钟，灭菌完成后待温度降至50℃时取出三角瓶并转移至洁净台，待温度降至室温时接种5-8块3-5mm菌丝块，然后180rpm避光培养30d，取出后用超声提取30min，然后使用3层无菌纱布过滤，得到发酵液。
醇提物制备方法：按照1：10的比例加入无水乙醇至发酵液中，4℃冰箱存放处理12h，然后50℃、40rpm条件下旋转蒸发减压浓缩，产物在-80℃、0.008mBAR条件下使用低温冷冻干燥机干燥，冻干完成后使用无菌密封袋低温保存。
方法：
1.消化对数生长期Du145，NIH3T3细胞，种24孔板，8000个/孔，每孔500μL；
2.各样品称量30mg溶于300μL DMSO,完全溶解后加300μL DMEM(10％FBS)，设置以下浓度100，250，300，350，400μg/ml，加药；
3.培养48h，观察细胞致死率，结果如表1所示。
表1细胞致死率结果
由表1结果可看出，发酵液的醇提物的IC50在300-350μg/mL之间，在浓度为400μg/mL时，Du145细胞死亡率为90％，NIH3T3细胞死亡率为100％，由此可说明，本发明所述菌根真菌的菌丝体发酵液醇提物能够有效抑制肿瘤细胞，可用于制备抗肿瘤药物等。
最后所应当说明的是，以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制，尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。
Claims (2)
1.一种菌根真菌，其特征在于，所述菌根真菌为伞菌目(Agaricales)口蘑科(Tricholomataceae)紫褶口蘑属(Tricholosporum)新菌株HMGIM-D160241,保藏于中国典型培养物保藏中心，保藏编号为：CCTCC M 2017185，保藏菌株号为HMGIM-D160241。
2.如权利要求1所述菌根真菌在制备抗肿瘤的药物中的用途。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710771269.3A CN107523506B (zh) | 2017-08-31 | 2017-08-31 | 一种菌根真菌及其在抗肿瘤中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710771269.3A CN107523506B (zh) | 2017-08-31 | 2017-08-31 | 一种菌根真菌及其在抗肿瘤中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107523506A CN107523506A (zh) | 2017-12-29 |
| CN107523506B true CN107523506B (zh) | 2019-09-17 |
Family
ID=60683030
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710771269.3A Active CN107523506B (zh) | 2017-08-31 | 2017-08-31 | 一种菌根真菌及其在抗肿瘤中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107523506B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108293599B (zh) * | 2018-02-28 | 2020-05-19 | 中华全国供销合作总社昆明食用菌研究所 | 一种花脸香蘑及其分离扩繁方法和覆土栽培方法 |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006133708A1 (en) * | 2005-06-15 | 2006-12-21 | Medimush A/S | Bioactive agents produced by submerged cultivation of a basidiomycete cell |
| CN1961652A (zh) * | 2005-11-07 | 2007-05-16 | 中国医学科学院药用植物研究所 | 菌根真菌在石斛大面积栽培中的应用 |
| CN1961651A (zh) * | 2005-11-07 | 2007-05-16 | 中国医学科学院药用植物研究所 | 菌根真菌在金线莲大面积栽培中的应用 |
| CN102220246A (zh) * | 2011-05-18 | 2011-10-19 | 辽宁省农业科学院 | 蓝莓菌根真菌（晶粒鬼伞）及其分离方法和用途 |
| CN103667084A (zh) * | 2013-12-19 | 2014-03-26 | 云南大学 | 使用菌根菌与腐生菌共同培养黄鳞鹅膏菌丝体的方法 |
| CN107090410A (zh) * | 2017-04-25 | 2017-08-25 | 鲁东大学 | 一株产植物激素的菌根真菌及其在促进植物生长中的应用 |
-
2017
- 2017-08-31 CN CN201710771269.3A patent/CN107523506B/zh active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006133708A1 (en) * | 2005-06-15 | 2006-12-21 | Medimush A/S | Bioactive agents produced by submerged cultivation of a basidiomycete cell |
| CN1961652A (zh) * | 2005-11-07 | 2007-05-16 | 中国医学科学院药用植物研究所 | 菌根真菌在石斛大面积栽培中的应用 |
| CN1961651A (zh) * | 2005-11-07 | 2007-05-16 | 中国医学科学院药用植物研究所 | 菌根真菌在金线莲大面积栽培中的应用 |
| CN102220246A (zh) * | 2011-05-18 | 2011-10-19 | 辽宁省农业科学院 | 蓝莓菌根真菌（晶粒鬼伞）及其分离方法和用途 |
| CN103667084A (zh) * | 2013-12-19 | 2014-03-26 | 云南大学 | 使用菌根菌与腐生菌共同培养黄鳞鹅膏菌丝体的方法 |
| CN107090410A (zh) * | 2017-04-25 | 2017-08-25 | 鲁东大学 | 一株产植物激素的菌根真菌及其在促进植物生长中的应用 |
Non-Patent Citations (1)
| Title |
|---|
| 松茸子实体活性多肽提取工艺;张倩等;《东北农业科学》;20170825;第42卷(第4期);第55-58页，参见全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107523506A (zh) | 2017-12-29 |
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 | |
| Khan et al. | Communities of endophytic fungi in medicinal plant Withania somnifera | |
| Roy et al. | Artificial cultivation of Ganoderma lucidum (Reishi medicinal mushroom) using different sawdusts as substrates | |
| Sridhar et al. | Evaluation of antibacterial and antifungal activity of Ganoderma lucidum (Curtis) P. Karst fruit bodies extracts | |
| CN101502218B (zh) | 桦褐孔菌菌核及子实体的人工栽培方法 | |
| CN103918472B (zh) | 牛樟芝的椴木栽培法 | |
| CN105039184A (zh) | 一种对波纹杂毛虫幼虫具有致病力的白僵菌菌株及其应用 | |
| CN108865949A (zh) | 枯草芽孢杆菌tkm-1及其应用 | |
| CN110042064B (zh) | 一种香菇菌株及其应用和源于该菌株的杀虫剂及其制备方法 | |
| KR20100104408A (ko) | 현미를 이용한 밀리타리스동충하초 균사체와 그를 이용한 건강보양용 밀타리스동충하초 삼계탕 | |
| CN107523506B (zh) | 一种菌根真菌及其在抗肿瘤中的应用 | |
| CN107811279A (zh) | 一种药食用真菌发酵核桃青皮的方法及其发酵制品与应用 | |
| Semwal et al. | Mushroom: Nature’s treasure in Ethiopia | |
| CN105803000A (zh) | 白鬼笔提取液、发酵胞外液和发酵胞内液的制备方法及其应用 | |
| CN107771613B (zh) | 一种黄褶裸伞新菌种及其栽培方法和用途 | |
| CN103285043B (zh) | 具有抗肿瘤功效的食用真菌蛋白质提取物的制备方法 | |
| CN107519212A (zh) | 一种菌根真菌子实体及其在抗肿瘤中的应用 | |
| KR101631926B1 (ko) | 버섯균사체 2차 배양을 통한 고품질 버섯균사체 대량 생산방법 | |
| Rajesh et al. | Phytochemical screening and biological activity of medicinal mushroom Ganoderma species | |
| Xhensila et al. | Mushroom Cultivation Systems: Exploring Antimicrobial and Prebiotic Benefits | |
| Kaur et al. | Evaluation of Ganoderma lucidum strains for the production of bioactive components and their potential use as antimicrobial agents | |
| JP6496959B2 (ja) | Ｏｐｈｉｏｃｏｒｄｙｃｅｐｓ属子実体形成用培地及びＯｐｈｉｏｃｏｒｄｙｃｅｐｓ属子実体の形成方法 | |
| CN110583363A (zh) | 采用代料栽培灵芝的工艺 | |
| Nasim et al. | Estimation of antimicrobial potential of Ganoderma lucidum (Leyss. Ex. Fr.) Karst. extracts | |
| KR102117227B1 (ko) | 까시구지뽕을 이용한 상황버섯 재배방법 및 그에 따른 상황버섯 |
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 | ||
| CP01 | Change in the name or title of a patent holder |
Address after: No. 100 courtyard of Yuexiu District martyr Middle Road, Yuexiu District, Guangzhou, Guangdong Co-patentee after: GUANGDONG YUEWEI EDIBLE FUNGI TECHNOLOGY Co.,Ltd. Patentee after: GUANGDONG INSTITUTE OF MICROBIOLOGY (GUANGDONG DETECTION CENTER OF MICROBIOLOGY) Address before: No. 100 courtyard of Yuexiu District martyr Middle Road, Yuexiu District, Guangzhou, Guangdong Co-patentee before: GUANGDONG YUEWEI EDIBLE FUNGI TECHNOLOGY Co.,Ltd. Patentee before: Guangdong Institute of Microbiology |
|
| CP01 | Change in the name or title of a patent holder | ||
| CP03 | Change of name, title or address |
Address after: 510070 No. 58, courtyard, No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou, Guangdong Province Patentee after: Institute of Microbiology, Guangdong Academy of Sciences Patentee after: GUANGDONG YUEWEI EDIBLE FUNGI TECHNOLOGY Co.,Ltd. Address before: 510075 courtyard, No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province Patentee before: GUANGDONG INSTITUTE OF MICROBIOLOGY (GUANGDONG DETECTION CENTER OF MICROBIOLOGY) Patentee before: GUANGDONG YUEWEI EDIBLE FUNGI TECHNOLOGY Co.,Ltd. |
|
| CP03 | Change of name, title or address | ||
| OL01 | Intention to license declared | ||
| OL01 | Intention to license declared |