CN112772297A - 一种臧氏牛肝菌菌种分离培养基 - Google Patents
一种臧氏牛肝菌菌种分离培养基 Download PDFInfo
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- 238000002955 isolation Methods 0.000 title claims abstract description 8
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims abstract description 36
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims abstract description 24
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims abstract description 24
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 24
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- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000004327 boric acid Substances 0.000 claims abstract description 12
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- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 claims abstract description 12
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 12
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 12
- 239000008103 glucose Substances 0.000 claims abstract description 12
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims abstract description 12
- 229960000367 inositol Drugs 0.000 claims abstract description 12
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims abstract description 12
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 12
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 12
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims abstract description 12
- 235000001968 nicotinic acid Nutrition 0.000 claims abstract description 12
- 229960003512 nicotinic acid Drugs 0.000 claims abstract description 12
- 239000011664 nicotinic acid Substances 0.000 claims abstract description 12
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000011780 sodium chloride Substances 0.000 claims abstract description 12
- RWVGQQGBQSJDQV-UHFFFAOYSA-M sodium;3-[[4-[(e)-[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]-2-methylcyclohexa-2,5-dien-1-ylidene]methyl]-n-ethyl-3-methylanilino]methyl]benzenesulfonate Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C(=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=2C(=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=C1 RWVGQQGBQSJDQV-UHFFFAOYSA-M 0.000 claims abstract description 12
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- 229960000344 thiamine hydrochloride Drugs 0.000 claims abstract description 12
- 235000019190 thiamine hydrochloride Nutrition 0.000 claims abstract description 12
- 239000011747 thiamine hydrochloride Substances 0.000 claims abstract description 12
- RZLVQBNCHSJZPX-UHFFFAOYSA-L zinc sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Zn+2].[O-]S([O-])(=O)=O RZLVQBNCHSJZPX-UHFFFAOYSA-L 0.000 claims abstract description 12
- 229920001817 Agar Polymers 0.000 claims abstract description 10
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 10
- 239000008272 agar Substances 0.000 claims abstract description 10
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 235000019838 diammonium phosphate Nutrition 0.000 claims abstract description 10
- 229910000388 diammonium phosphate Inorganic materials 0.000 claims abstract description 10
- 239000003102 growth factor Substances 0.000 claims abstract description 10
- 235000010333 potassium nitrate Nutrition 0.000 claims abstract description 10
- 239000004323 potassium nitrate Substances 0.000 claims abstract description 10
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229960004172 pyridoxine hydrochloride Drugs 0.000 claims abstract description 10
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 claims abstract description 10
- 239000011764 pyridoxine hydrochloride Substances 0.000 claims abstract description 10
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 9
- 239000000843 powder Substances 0.000 claims abstract description 9
- BDOYKFSQFYNPKF-UHFFFAOYSA-N 2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid;sodium Chemical compound [Na].[Na].OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O BDOYKFSQFYNPKF-UHFFFAOYSA-N 0.000 claims abstract description 7
- 150000003839 salts Chemical class 0.000 claims abstract description 7
- 239000002609 medium Substances 0.000 claims abstract description 6
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 claims description 9
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 8
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 8
- 239000004615 ingredient Substances 0.000 claims description 6
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- 159000000014 iron salts Chemical class 0.000 claims description 2
- 241000590002 Helicobacter pylori Species 0.000 claims 1
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- 241000233866 Fungi Species 0.000 abstract description 11
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- 150000002505 iron Chemical class 0.000 abstract 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 abstract 1
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- DBPRUZCKPFOVDV-UHFFFAOYSA-N Clorprenaline hydrochloride Chemical compound O.Cl.CC(C)NCC(O)C1=CC=CC=C1Cl DBPRUZCKPFOVDV-UHFFFAOYSA-N 0.000 description 2
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明属于微生物技术领域,具体涉及一种臧氏牛肝菌菌种分离培养基,包含大量成分,微量成分,铁盐和生长因子。大量成分包含特定浓度的葡萄糖、麦芽浸粉、无水氯化钙、无水硫酸镁、磷酸二氢钾、磷酸氢二铵、硝酸铵硝酸钾、琼脂;微量成分包含特定浓度的氯化钠、硼酸、一水合硫酸锰、七水合硫酸锌、碘化钾、二水合钼酸钠、五水合硫酸铜、六水合氯化钴;铁盐包含特定浓度的氯化铁、无水硫酸亚铁、乙二胺四乙酸二钠;生长因子包含特定浓度的肌醇、烟酸、盐酸吡哆醇、盐酸硫胺素、甘氨酸。该培养基有利于臧氏牛肝菌菌株生长,菌株活力高,生长速度快,产生的气生菌丝良好,为共生真菌的分子生物学研究和外生菌根真菌的栽培提供了基础保障。
Description
技术领域
本发明属于微生物技术领域,具体涉及一种臧氏牛肝菌菌种分离培养基。
背景技术
褐盖臧氏牛肝菌属于牛肝菌科臧氏牛肝菌属的物种,能与植物形成外生菌根,是一种共生真菌,是研究植物与真菌共生关系的理想材料,分离培养臧氏牛肝菌菌种有利于开展后续分子生物学实验操作,并为外生菌根真菌的栽培提供可能的基础保障。臧氏牛肝菌菌种分离培养的难点在于菌丝易老化,不能获得足够的气生菌丝。
一直以来,共生真菌的分离培养是比较困难的,不同的共生真菌对培养基有不同的要求。通用的共生真菌培养基为MMN培养基,腐生真菌培养基为PDA培养基,但实验发现,使用这两种培养基分离培养臧氏牛肝菌菌种,长势欠佳,菌丝易老化,气生菌丝较少,不适合共生牛肝菌菌种的生长,阻碍了后续研究工作的开展。目前也没有专门针对共生臧氏牛肝菌的培养基配方。
发明内容
为解决上述现有技术的不足,本发明提供一种臧氏牛肝菌菌种分离培养基,使臧氏牛肝菌菌株较长时间保持活力,产生良好的气生菌丝。
为实现上述目的,本发明采用的技术方案为:
一种臧氏牛肝菌菌种分离培养基,包含大量成分,微量成分,铁盐和生长因子,其中,所述的大量成分包含浓度为4000-6000mg/L葡萄糖、1000-2000mg/L麦芽浸粉、180-190mg/L无水氯化钙、120-135mg/L无水硫酸镁、320-350mg/L磷酸二氢钾、115-135mg/L磷酸氢二铵、815-835mg/L硝酸铵、940-960mg/L硝酸钾、12000-16000mg/L琼脂;所述微量成分包含浓度为11.5-13.5mg/L氯化钠、3.0-3.2mg/L硼酸、8.2-8.6mg/L一水合硫酸锰、4.1-4.5mg/L七水合硫酸锌、0.400-0.430mg/L碘化钾、0.100-0.140mg/L二水合钼酸钠、0.0115-0.0135mg/L五水合硫酸铜、0.0115-0.0135mg/L六水合氯化钴;所述铁盐包含浓度为4-8mg/L氯化铁、7.0-8.0mg/L无水硫酸亚铁、18.4-18.8mg/L乙二胺四乙酸二钠;所述生长因子包含浓度为40-60mg/L肌醇、0.15-0.35mg/L烟酸、0.15-0.35mg/L盐酸吡哆醇、0.05-0.15mg/L盐酸硫胺素、0.6-1.5mg/L甘氨酸。
作为优选的,所述的大量成分包含浓度为4500-5500mg/L葡萄糖、1200-1800mg/L麦芽浸粉、182-186mg/L无水氯化钙、125-130mg/L无水硫酸镁、330-340mg/L磷酸二氢钾、120-130mg/L磷酸氢二铵、820-830mg/L硝酸铵、945-955mg/L硝酸钾、13000-15000mg/L琼脂;所述微量成分包含浓度为12.0-13.0mg/L氯化钠、3.05-3.15mg/L硼酸、8.4-8.5mg/L一水合硫酸锰、4.2-4.4mg/L七水合硫酸锌、0.410-0.420mg/L碘化钾、0.120-0.130mg/L二水合钼酸钠、0.012-0.013mg/L五水合硫酸铜、0.012-0.013mg/L六水合氯化钴;所述铁盐包含浓度为5-7mg/L氯化铁、7.4-7.8mg/L无水硫酸亚铁、18.6-18.7mg/L乙二胺四乙酸二钠;所述生长因子包含浓度为45-55mg/L肌醇、0.2-0.3mg/L烟酸、0.2-0.3mg/L盐酸吡哆醇、0.08-0.12mg/L盐酸硫胺素、0.8-1.3mg/L甘氨酸。
作为优选的,所述的大量成分包含浓度为5000mg/L葡萄糖、1500mg/L麦芽浸粉、184mg/L无水氯化钙、127.5mg/L无水硫酸镁、335mg/L磷酸二氢钾、125mg/L磷酸氢二铵、825mg/L硝酸铵、950mg/L硝酸钾、14000mg/L琼脂;所述微量成分包含浓度为12.5mg/L氯化钠、3.1mg/L硼酸、8.45mg/L一水合硫酸锰、4.3mg/L七水合硫酸锌、0.415mg/L碘化钾、0.125mg/L二水合钼酸钠、0.0125mg/L五水合硫酸铜、0.0125mg/L六水合氯化钴;所述铁盐包含浓度为6mg/L氯化铁、7.6mg/L无水硫酸亚铁、18.63mg/L乙二胺四乙酸二钠;所述生长因子包含浓度为50mg/L肌醇、0.25mg/L烟酸、0.25mg/L盐酸吡哆醇、0.1mg/L盐酸硫胺素、1mg/L甘氨酸。
本发明的培养基进行臧氏牛肝菌菌种分离培养,菌株活力高,生长速度较快,菌株菌团直径大,产生的气生菌丝良好,为共生真菌的分子生物学研究和外生菌根真菌的栽培提供了基础保障。
具体实施方式
实施例1
称取氯化钠2.3g、硼酸0.6g、一水合硫酸锰1.64g、七水合硫酸锌0.82g、碘化钾80mg、二水合钼酸钠20mg、五水合硫酸铜2.3mg、六水合氯化钴2.3mg,用少量蒸馏水溶解后定容至1L,保存于棕色玻璃瓶中,标记为母液;
称取葡萄糖4g、麦芽浸粉1g、无水氯化钙0.18g、无水硫酸镁0.12g、磷酸二氢钾0.32g、磷酸氢二铵0.115g、硝酸铵0.815g、硝酸钾0.940g、琼脂12g,置于1L的三角瓶中,分别从母液、、中吸取5ml溶液置于上述三角瓶中,用少量蒸馏水溶解后定容至1L,再平均分装到两个1L的三角瓶中,密封后置于灭菌锅(103.4KPa,121℃)中进行灭菌20分钟,取出待温度降至40-50℃时,将培养基倒入培养皿中,凝固后灭菌(压力0.105MPa , 121℃,灭菌15-30mⅠn)即可进行菌种分离培养操作。
实施例2
称取氯化钠2.7g、硼酸0.64g、一水合硫酸锰1.72g、七水合硫酸锌0.9g、碘化钾80mg、二水合钼酸钠20mg、五水合硫酸铜2.3mg、六水合氯化钴2.3mg,用少量蒸馏水溶解后定容至1L,保存于棕色玻璃瓶中,标记为母液;
称取葡萄糖6g、麦芽浸粉2g、无水氯化钙0.19g、无水硫酸镁0.135g、磷酸二氢钾0.35g、磷酸氢二铵0.135g、硝酸铵0.835g、硝酸钾0.960g、琼脂16g,置于1L的三角瓶中,分别从母液、、中吸取5ml溶液置于上述三角瓶中,用少量蒸馏水溶解后定容至1L,再平均分装到两个1L的三角瓶中,密封后置于灭菌锅(103.4KPa,121℃)中进行灭菌20分钟,取出待温度降至40-50℃时,将培养基倒入培养皿中,凝固后灭菌(压力0.105MPa , 121℃,灭菌15-30mⅠn)即可进行菌种分离培养操作。
实施例3
称取氯化钠2.4g、硼酸0.61g、一水合硫酸锰1.68g、七水合硫酸锌0.84g、碘化钾82mg、二水合钼酸钠24mg、五水合硫酸铜2.4mg、六水合氯化钴2.4mg,用少量蒸馏水溶解后定容至1L,保存于棕色玻璃瓶中,标记为母液;
称取葡萄糖4.5g、麦芽浸粉1.2g、无水氯化钙0.182g、无水硫酸镁0.125g、磷酸二氢钾0.33g、磷酸氢二铵0.120g、硝酸铵0.820g、硝酸钾0.945g、琼脂13g,置于1L的三角瓶中,分别从母液、、中吸取5ml溶液置于上述三角瓶中,用少量蒸馏水溶解后定容至1L,再平均分装到两个1L的三角瓶中,密封后置于灭菌锅(103.4KPa,121℃)中进行灭菌20分钟,取出待温度降至40-50℃时,将培养基倒入培养皿中,凝固后灭菌(压力0.105MPa ,121℃,灭菌15-30mⅠn)即可进行菌种分离培养操作。
实施例4
称取氯化钠2.6g、硼酸0.63g、一水合硫酸锰1.70g、七水合硫酸锌0.88g、碘化钾84mg、二水合钼酸钠26mg、五水合硫酸铜2.6mg、六水合氯化钴2.6mg,用少量蒸馏水溶解后定容至1L,保存于棕色玻璃瓶中,标记为母液;
称取葡萄糖5.5g、麦芽浸粉1.8g、无水氯化钙0.186g、无水硫酸镁0.13g、磷酸二氢钾0.34g、磷酸氢二铵0.130g、硝酸铵0.830g、硝酸钾0.955g、琼脂15g,置于1L的三角瓶中,分别从母液、、中吸取5ml溶液置于上述三角瓶中,用少量蒸馏水溶解后定容至1L,再平均分装到两个1L的三角瓶中,密封后置于灭菌锅(103.4KPa,121℃)中进行灭菌20分钟,取出待温度降至40-50℃时,将培养基倒入培养皿中,凝固后灭菌(压力0.105MPa , 121℃,灭菌15-30mⅠn)即可进行菌种分离培养操作。
实施例5
称取氯化钠2.5g、硼酸0.62g、一水合硫酸锰1.69g、七水合硫酸锌0.86g、碘化钾83mg、二水合钼酸钠25mg、五水合硫酸铜2.5mg、六水合氯化钴2.5mg,用少量蒸馏水溶解后定容至1L,保存于棕色玻璃瓶中,标记为母液;
称取葡萄糖5.0g、麦芽浸粉1.5g、无水氯化钙0.184g、无水硫酸镁0.1275g、磷酸二氢钾0.335g、磷酸氢二铵0.125g、硝酸铵0.825g、硝酸钾0.950g、琼脂14g,置于1L的三角瓶中,分别从母液、、中吸取5ml溶液置于上述三角瓶中,用少量蒸馏水溶解后定容至1L,再平均分装到两个1L的三角瓶中,密封后置于灭菌锅(103.4KPa,121℃)中进行灭菌20分钟,取出待温度降至40-50℃时,将培养基倒入培养皿中,凝固后灭菌(压力0.105MPa ,121℃时,灭菌15-30mⅠn)后待用。
采用PDA、MMN和本发明的培养基,按照以下步骤进行臧氏牛肝菌菌种分离培养:①采集臧式牛肝菌的新鲜子实体,去除表面杂物,于超净实验台中,用手将子实体一分为二;②将手术刀和镊子在酒精灯上灭菌冷却,用手术刀在子实体的菌盖菌肉中心部位划取约2-3mm大小的组织块,并用镊子将组织块放入盛有不同培养基的培养皿中,每个培养皿中放置3个组织块;③将上述培养皿置于22℃恒温培养箱中进行培养4-5周,再划取2-3mm大小且长有菌丝的培养基块,放至新的培养皿中,每个培养皿中放入三块,再放入22℃恒温培养箱中进行培养,观察菌丝成长情况并测量菌丝团直径。三种培养基下臧氏牛肝菌菌丝生长情况见表1。从对比结果看,本发明的培养基进行臧氏牛肝菌菌种培养,菌丝团直径,产生的气生菌菌丝均优于其他两种培养基。
表1 不同菌种分离培养基下臧氏牛肝菌菌丝生长情况(13周)
Claims (3)
1.一种臧氏牛肝菌菌种分离培养基,包含大量成分,微量成分,铁盐和生长因子,其特征在于,所述的大量成分包含浓度为4000-6000mg/L葡萄糖、1000-2000mg/L麦芽浸粉、180-190mg/L无水氯化钙、120-135mg/L无水硫酸镁、320-350mg/L磷酸二氢钾、115-135mg/L磷酸氢二铵、815-835mg/L硝酸铵、940-960mg/L硝酸钾、12000-16000mg/L琼脂;所述微量成分包含浓度为11.5-13.5mg/L氯化钠、3.0-3.2mg/L硼酸、8.2-8.6mg/L一水合硫酸锰、4.1-4.5mg/L七水合硫酸锌、0.400-0.430mg/L碘化钾、0.100-0.140mg/L二水合钼酸钠、0.0115-0.0135mg/L五水合硫酸铜、0.0115-0.0135mg/L六水合氯化钴;所述铁盐包含浓度为4-8mg/L氯化铁、7.0-8.0mg/L无水硫酸亚铁、18.4-18.8mg/L乙二胺四乙酸二钠;所述生长因子包含浓度为40-60mg/L肌醇、0.15-0.35mg/L烟酸、0.15-0.35mg/L盐酸吡哆醇、0.05-0.15mg/L盐酸硫胺素、0.6-1.5mg/L甘氨酸。
2.根据权利要求1所述的一种臧氏牛肝菌菌种分离培养基,其特征在于,所述的大量成分包含浓度为4500-5500mg/L葡萄糖、1200-1800mg/L麦芽浸粉、182-186mg/L无水氯化钙、125-130mg/L无水硫酸镁、330-340mg/L磷酸二氢钾、120-130mg/L磷酸氢二铵、820-830mg/L硝酸铵、945-955mg/L硝酸钾、13000-15000mg/L琼脂;所述微量成分包含浓度为12.0-13.0mg/L氯化钠、3.05-3.15mg/L硼酸、8.4-8.5mg/L一水合硫酸锰、4.2-4.4mg/L七水合硫酸锌、0.410-0.420mg/L碘化钾、0.120-0.130mg/L二水合钼酸钠、0.012-0.013mg/L五水合硫酸铜、0.012-0.013mg/L六水合氯化钴;所述铁盐包含浓度为5-7mg/L氯化铁、7.4-7.8mg/L无水硫酸亚铁、18.6-18.7mg/L乙二胺四乙酸二钠;所述生长因子包含浓度为45-55mg/L肌醇、0.2-0.3mg/L烟酸、0.2-0.3mg/L盐酸吡哆醇、0.08-0.12mg/L盐酸硫胺素、0.8-1.3mg/L甘氨酸。
3.根据权利要求1所述的一种臧氏牛肝菌菌种分离培养基,其特征在于,所述的大量成分包含浓度为5000mg/L葡萄糖、1500mg/L麦芽浸粉、184mg/L无水氯化钙、127.5mg/L无水硫酸镁、335mg/L磷酸二氢钾、125mg/L磷酸氢二铵、825mg/L硝酸铵、950mg/L硝酸钾、14000mg/L琼脂;所述微量成分包含浓度为12.5mg/L氯化钠、3.1mg/L硼酸、8.45mg/L一水合硫酸锰、4.3mg/L七水合硫酸锌、0.415mg/L碘化钾、0.125mg/L二水合钼酸钠、0.0125mg/L五水合硫酸铜、0.0125mg/L六水合氯化钴;所述铁盐包含浓度为6mg/L氯化铁、7.6mg/L无水硫酸亚铁、18.63mg/L乙二胺四乙酸二钠;所述生长因子包含浓度为50mg/L肌醇、0.25mg/L烟酸、0.25mg/L盐酸吡哆醇、0.1mg/L盐酸硫胺素、1mg/L甘氨酸。
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