The Prevention and Treatment of Polysaccharide from the Rhizomorph of Armillaria mellea on Diabetic Cataract in Rat

[Objective] To investigate the prevention and control effect of polysaccharide from the rhizomorph of Armillaria mellea by distilled water elution （AMP-1） on diabetic cataract in rats.[Methods] Streptozotocin （STZ）,combined with freund＇s adjuvant （CFA）,was used to induce diabetic cataract in rat,to observe the onset of lenticular turbidity by slit lamp and compare the turbid degree of eyes lens among different groups.The super oxide dismutase （SOD）,glutathione peroxidase （GSH-px） and malondialdehyde （MDA） in serum and eyes lens were measured by colorimetric method.[Results] The slit lamp examination results showed that AMP-1 obviously delayed the onset of diabetic cataract and reduced turbid degree of eyes lens.Compared with model group,AMP-1 at various doses should significantly reduce the level of MDA ascended in serum of rats.Medium-and high-dose groups could increase the levels of SOD and GSH-Px in serum,and the dose of AMP-1 was closely related to the effect.[Conclusion] AMP-1 played a prominent role in prevention and treatment of diabetic cataract in rats.

Carbon-13 Nuclear Magnetic Resonance Spectra of Some Protoilludane Sesquiterpenoid Aromatic Esters from Armillaria mellea (Vahl.ex Fr.) Quel.

The carbon-13 nuclear magnetic resonance spectra of seventeen protoilludane ses- quiterpenoid aromatic esters from the artificially cultured mycelium of Armillaria mellea (Vahl.ex Fr.) Quel.have been analysed and assigned.All the ^(11)CNMR signals of the model compounds melleolide 5 and compounds 8,10,11,13,14,15,and 16 can be assigned on the basis of the multiplicity of the sig- nals in the off-resonance decoupled spectra or INEPT spectra.^(11)C-~1H COSY and long-range ^(13)C-~1H COSY.The assignments of the ^(13)CNMR spectra of other protoilludane sesquiterpenoid aromatic esters were completed by comparison with the model compounds as well as the electronic effects of substi- tuted group on the molecules.The present results indicate that ^(11)CNMR spectroscopy provides an ef- fective method for characterizing the protoilludane sesquiterpenoid aromatic esters.

野生蜜环菌(Armillaria Mellea)的分离纯化及抑菌作用

分离纯化野生蜜环菌菌丝培养物,研究菌丝体代谢物质对大肠杆菌、金黄葡萄球菌、志贺氏菌和副溶血性弧菌的抑制作用.采集野生蜜环菌子实体,获得遗传同一性菌丝体纯培养物,振荡培养获得菌丝体代谢液,观察其对致病菌的抑菌效果.实验结果表明,分离纯化得到的菌丝体纯培养物与蜜环菌遗传同一,菌丝体代谢液对大肠杆菌、金黄葡萄球菌、志贺氏菌和副溶血性弧菌等食品中常见的致病菌具有较好的抑制作用。

A New Ceramide from the Basidiomycetes Armillaria mellea

A new phytosphingosine-type ceramide, armillaramide 1, has been isolated from the fruiting bodies of Basidiomycetes Armillaria mellea (Vahl ex Fr.) Quel. Its structure was established as (2S, 3S. 4R)-2-N-(palmitoyl)-phytosphingosine by spectroscopic and chemical methods.

THE STRUCTURE OF ARMILLARIZIN,A NEW PROTOILLUDANE SESQUITERPENOID AROMATIC ESTER FROM ARMILLARIA MELLEA(VAHL.EX FR)QUEL.

A new protoilludane sesquiterpenoid aromatic ester,named armillarizin (1)has been isolated from the artificially cultured mycelium of Armillaria me- llea(Vahl.ex Fr)Quel.(Tricholomataceae).Its structure was deduced on the ba- sis of spectral analyses.The relative configuration was established from the nuclear Overhauser effects and from the values of ~1H-~1H coupling constants.

Screening of Semi-solid Culture Conditions of Armillaria mellea A9

[Objectives]This study was conducted to optimize the germplasm resources of Gastrodia elata and improve the condition of declining G.elata production.[Methods]A scientific and reasonable method was used to screen the optimum nutritional conditions(carbon sources,nitrogen sources,microelements,vitamins)and environmental conditions(pH,temperature,light)of Armillaria mellea A9 growth,so as to screen the medium suitable for the growth of A.mellea.[Results]Under the semi-solid culture condition,the suitable pH range of A.mellea mycelia was 4.0-9.0;the temperature was about 25℃;and the light condition was dark culture.Under the experimental conditions,the optimal carbon source was 25 g/L ethanol;the optimal nitrogen source was 2.5 g/L soy peptone;and the optimum microelement and vitamin were 0.5 g/L MnSO 4,0.5 g/L MgSO 4 and 20 mg/L vitamin B6,respectively.After screening,the growth rate of A.mellea rhizomorph was as high as 13.52 mm/d,and the dry weight of mycelium also reached 0.296 7 g.[Conclusions]This study not only omitted a lot of tedious experiments,but also obtained reliable and scientific experimental results,and achieved the purpose of rejuvenating A.mellea,which is conducive to the further development and utilization of G.elata and its symbiotic fungi.

Chemical Constituent of Natural Body of Armillaria mellea and Structure of 5,6-Epoxy-22-en-3-ol-ergosterol

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Chemical Compositions of Volatile Oil from Fruiting Body of Armillaria luteo-virens

The chemical compositions of volatile oil from fruiting body of Armillaria luteo-virens in Qinghai Province were firstly analyzed with GC-MS and its relevant compositions were detected by calculating chromatographic peak area with normalized method. 21 peaks were separated and 13 compositions were identified which were mainly unsaturated fatty acids, taking 97.1% of the total volatile oil.

Simple Culture Techniques for Germinationpromoting Fungi and Armillariella mellea in Artificial Planting of Gastrodia elata

This study aimed to explore simple culture techniques for symbiotic germination-promoting fungi and Armillariella mellea in artificial planting of Gastrodia elata, i.e., how to use a corner of ordinary house as inoculation room and cultivating site, how to use a pressure cooker for cooking food as sterilization tool, and how to use ordinary household heating as heating measures.

Screening of Strains and Nutrition Sources for Culture of Armillariella mellea Rhizomoph Branch in Ganzi

Based on the natural resources in Ganzi Tibetan Autonomous Prefecture, screening experiment of strains and nutrient sources for culture of Armillariella mellea rhizomoph branch was carried out with 3 strains of A. mellea, 2 types of main nutrient sources and 3 kinds of auxiliary nutrient sources. Total 18 treatments were designed. The results showed that with the A. mellea strain separated from the tubers of wild Gastrodia elata BI. by tissue isolation as mother strain, the shrubs such as Prunus tatsienensis Batal and Quercus spinosa David ex Franch as the main nutrient sources, and the agricultural products such as corn and potato as the supplementary nutrient sources, the growth of A. mellea rhizomoph branch was best in bottle.

Structures and cytotoxicities of three new sesquiterpenes from cultures of Armillaria sp.

Three new sesquiterpene aryl esters,named 10-dehydroxy-melleoliede B(1),1-O-formyl-10-dehydroxy-melleoliede B(2)and 10-oxo-melleoliede B(3)together with six known ones(4-9),were isolated from the cultures of Armillaria sp.The structures of the new compounds were elucidated based on the extensive spectroscopic methods.Compounds 1,2,and 5-9 exhibited moderate cytotoxicities.

ARMILLARIA TRAPPING IN A PINUS KORAIENSIS PLANTATION IN NORTHEAST CHINA

Trap bags were used to assess the presence of an Armillaria species in a Pinus koraiensis plantation in Heilongjiang, China. 216 trap bags containing bark were placed in three plots at 2×2m spacing. After 10 weeks, the presence of Armillaria was determined from the growth of rhizomorphs in the trap bags. The trap bag method proved successful in determining the presence of viable Armillaria rhizomorphs where only a few dead trees showed signs of the disease. The distribution of positive traps corresponded generally with the presence of rhizomorphs in the soil taken from the trap holes. However, the trap bags were superior to soil samples for evaluating the presence of viable rhizomorphs. The fresh, abundant rhizomorphs were easily identified by persons inexperienced in the assessment of Armillaria root rot. Several practical problems need to be solved before the method is applicable in general forest management in China.

Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(银杏蜜环口服溶液) attenuates inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis in post stroke depression

OBJECTIVE The Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(Yinxingmihuan Koufu Rongye,YXMH),a representative drug for"Treating both Brain and Heart",showed considerable clinical effects in isch⁃emic cardiovascular and cerebral vascular diseases.Recently,it is reported that YXMH has the potential for treating myocardial and cerebral ischemia related mental disorders,such as post stroke depression(PSD)and chronic heart disease(CHD)associated anxiety disorder.However,its mechanism has not been clearly elucidated.Meanwhile,increasing evidence revealed that there are close functional links between depression and habenular nucleus.The present study investigates the underlying mechanism of YXMH on attenuating the inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis in in a rat model of PSD.METHODS Rats were randomly devided into sham group,model group,Ginaton group(18 mg·kg^-1),Armillariella Mellea group(600 mg·kg-1),Fluoxetine group(10 mg·kg^-1),YXMH high-dose group(618 mg·kg^-1)and YXMH low-dose group(309 mg·kg^-1).The PSD model was induced by transarterial microembolization combined with sleep deprivation(2-Chloro-D-phenylalanine,PCPA,IH,200 mg·kg^-1,for 3 times,before the behavior test)in SD male rats.Then rats were treated with corresponding medicaments through gavage once a day until 3 weeks later,followed by body mass measurement,neurological deficit score evaluation,gripping strength and thermal withdrawl latency measurement,as well as depression related behavioral indicators,the open field test(OFT)and sucrose preference test.The pathological morphological changes of habenular nucleus was observed by HE staining,the expression of IBA-1 was measured and analyzed by immunohistochemistry staining,and alterations of proteins and genes related to the CX3CL1-CX3CR1 axis were analyzed using Western blotting(CX3CL1,CX3CR1)and real-time polymerase chain reaction(PCR)(CX3CL1,CX3CR1).RESULTS Compared with the sham group,rats in the model group manifested as decreased body mass,deficient neurological behavior and gripping strength,reduced loco⁃motor activity and sugar water consumption,as well as elevated thermal withdrawl latency(P<0.05,P<0.01).Mean⁃while,the pathological morphology of the habenular nucleus on the ischemic hemisphere showed significant neuronal degeneration,microglial proliferation,inflammatory cells and glia cells infiltration,together with up-regualted expression of IBA-1,CX3CL1,CX3CR1 protein and CX3CL1,CX3CR1 mRNA.YXMH attenuated inflammation of microglia in habenular nucleus through improving pathological morphology,inhibiting IBA-1 activation,down-regulating the expres⁃sion of CX3CL1 and CX3CR1 proteins and genes,and thus improved the behavior performance of ischemic injury and depression.CONCLUSION YXMH ameliorates neurological deficit and depressive behavior in rat model of PSD induced by transarterial microembolization combined with sleep deprivation,and the mechanism is probably related to attenu⁃ating inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis.

蜜环菌人工菌棒基质碳氮比对乌天麻产量及营养品质的影响

目的分析蜜环菌人工菌棒基质碳氮比对乌天麻产量及基本营养成分含量的影响,并对乌天麻氨基酸品质进行评价。方法通过大棚内随机区组实验,测定不同种植模式下乌天麻的基本营养成分含量,对其进行单因素方差分析和多重比较,通过氨基酸评分法对乌天麻中氨基酸营养价值进行评价。结果蜜环菌人工菌棒基质碳氮比对乌天麻的产量、基本营养成分含量及氨基酸的营养品质均有显著影响;蜜环菌人工菌棒基质碳氮比为75.94时种植的乌天麻,其各项营养指标接近对照种植的乌天麻的各项营养指标,显著大于蜜环菌人工菌棒基质碳氮比为60.69、126.39时种植的乌天麻的各项营养指标。蜜环菌人工菌棒基质碳氮比为75.94时种植的乌天麻,其必需氨基酸含量与总氨基酸含量的比值为37.03%,接近40%;必需氨基酸含量与非必需氨基酸含量的比值为58.79%,接近60%;必需氨基酸的氨基酸比值、必需氨基酸的比值系数分、必需氨基酸比值系数与联合国粮食及农业组织/世界卫生组织提出的氨基酸理想模式较接近。结论利用蜜环菌人工菌棒种植乌天麻可以获得基本营养成分含量高、氨基酸丰富、必需氨基酸的组成比例相对合理、符合人体吸收利用需求、具有较高营养价值的乌天麻。

QUANTITATIVE ANALYSIS OF RELATIONSHIP BETWEEN ARMILLARIA ROOT ROT AND SITE FACTORS IN PINUS RADIATA PLANTATIONS

Incidence of Armillaria infection was quantified based on site factors in New Zealand Pinus radiata plantations.A linear multiple regression model was derived to predict infection levels of Armillaria root rot.Factors positivily associated with the infection were:previous vegetation（native bush,pine）;soil type（pumice）;landform （valley,gully,flat）and the interaction between them.This model could assist in management planning with regard to the predisposition of particular stand to Armillaria infection.Keywods:Armillaria root rot,Disease incidence,Site factors,Quantification,Pinus radiata.

土壤相对含水量对乌天麻生物学效率及营养品质的影响

以乌天麻为试材,采用氨基酸评分法,研究了土壤相对含水量(RWC)对蜜环菌人工菌棒种植乌天麻的生物学效率(BE)及营养品质的影响,以期为天麻产业健康发展提供依据。结果表明:1)采用蜜环菌人工菌棒种植乌天麻,土壤RWC对乌天麻的BE、基本营养成分含量及氨基酸的营养品质均有显著影响,土壤RWC为75%时,乌天麻的BE为28.74%,显著高于对照(CK)及土壤RWC为55%、65%、85%时;土壤RWC为75%时,乌天麻的基本营养成分含量与CK接近,显著大于土壤RWC为55%、65%、85%时(P<0.05);2)在乌天麻中共检测到15种氨基酸,包括6种人体必需氨基酸(EAA)和9种人体非必需氨基酸(NEAA),当土壤RWC为75%时,利用蜜环菌人工菌棒种植的乌天麻和CK的乌天麻其EAA/TAA比值、EAA/NEAA比值、必需氨基酸的氨基酸比值(AAS)、必需氨基酸比值系数(RC)、必需氨基酸的比值系数分(SRC)与联合国粮食及农业组织/世界卫生组织(FAO/WHO)提出的氨基酸理想模式较接近;3)利用蜜环菌人工菌棒种植乌天麻,通过控制土壤RWC,可以获得必需氨基酸的组成比例相对合理,营养价值均衡,具有较高营养价值的乌天麻。

蜜环菌发酵玉米醇溶蛋白中蛋白酶学性质研究

为探究大型食药用真菌胞外蛋白酶的相关特性,从蜜环菌发酵的玉米醇溶蛋白产物中分离、纯化蛋白酶,通过饱和硫酸铵沉淀粗酶液、DEAE Sepharose Fast Flow阴离子层析柱及Sephadex G-50层析柱纯化,得到电泳纯的蛋白酶。结果表明,该蛋白酶的分子质量为18.4 ku;最适温度为60℃,在40~60℃具有良好稳定性;pH值为8时酶活力最高,pH值稳定性范围为6~8;Mn2+对该酶具有明显的促进作用,乙二胺四乙酸(EDTA)对中性蛋白酶有明显的抑制作用;米氏常数Km为3.786 mg/mL。研究结果为大型食药用真菌来源蛋白酶的研究与应用提供理论依据,为大型真菌源酶制剂的研发提出新思路。

静磁场对蜜环菌发酵产物结构和理化特性的影响

采用蜜环菌普通发酵及静磁场辅助发酵玉米醇溶蛋白,探究发酵期间静磁场对玉米醇溶蛋白发酵产物结构和理化特性的影响。傅里叶变换红外光谱分析发现静磁场处理改变了发酵产物蛋白的二级结构;紫外吸收光谱和内源荧光光谱分析表明,经静磁场处理后产物蛋白分子结构舒展、基团暴露;扫描电子显微镜结果显示静磁场处理后产物蛋白表面更加粗糙,出现孔洞,比表面积增大;热力学性质分析结果显示静磁场处理后产物蛋白的变性温度明显升高,且静磁场处理使粒径减小,Zeta电位绝对值增大,游离巯基含量呈现上升趋势,二硫键含量呈现下降趋势;此外,静磁场辅助发酵明显改变了产物蛋白的理化特性,持水性、持油性、乳化性、起泡性和泡沫稳定性均明显升高,在第7天时达到最大,分别为3.86 g/g、4.22 g/g、26.83 m^(2)/g、37.67%和58.12%,乳化稳定性明显降低,在第7天时最小为34.10%。综上,静磁场辅助蜜环菌发酵玉米醇溶蛋白能够改变产物蛋白的结构,并有效改善其理化特性,进一步提高玉米醇溶蛋白的附加值和利用率。

蜜环菌Am-07-22发酵对玉米赤霉烯酮降解效果影响及机理初探

本研究以蜜环菌Am-07-22为试验菌株,采用真菌生物发酵的方式降解玉米赤霉烯酮(ZEN),对蜜环菌降解ZEN的降解效果进行研究,包括菌株对不同浓度ZEN的降解效果以及不同培养时间、培养温度、初始pH和接种量对菌株降解ZEN的影响。然后对降解机理进行初探,分析了菌丝体、发酵上清液、细胞内容物对ZEN的降解作用,并研究了不同发酵时间、pH、金属离子对发酵上清液降解ZEN的影响,以及降解效果与菌株产漆酶活力的相关性分析。结果表明:蜜环菌Am-07-22对ZEN的降解效果良好,当ZEN浓度为5μg/mL时,最适降解条件为培养时间8 d,培养温度27℃,初始pH7.0,接种量10%,此时对ZEN的降解率为78.72%。菌丝体、发酵上清液和细胞内容物对ZEN的降解率分别为47.42%、37.05%和13.08%,蜜环菌Am-07-22分泌的胞外酶是降解ZEN的主要方式,而且菌体细胞对ZEN也有一定的吸附作用。另外,发酵上清液对ZEN的降解率与漆酶酶活的相关性较高为0.973,且Cu^(2+)对发酵上清液降解ZEN具有最佳的促进作用。

青海黄蘑菇色素提取及相关性质研究

为明确黄蘑菇色素的理化性质和抗氧化活性,本研究采用响应面设计优化了黄蘑菇色素提取工艺,研究了色素稳定性和抗氧化活性。结果表明,黄蘑菇色素最佳提取工艺条件为去离子水作为提取剂,提取温度51℃,提取时间61 min,料液比1∶113 g·mL^(-1),提取率为90.93%。黄蘑菇色素粗提物对光比较敏感,避光条件下更稳定,耐高温和耐还原性较差,但其具有一定的耐酸碱性和耐氧化性;Zn^(2+)的添加增大了黄蘑菇色素的吸光度,Al^(3+)浓度超过0.25%时,色素吸光度下降;葡萄糖对色素稳定性影响较小;苯甲酸钠和蔗糖的浓度超过1%时,色素吸光度增加,对其稳定性影响较大。色素粗提物对1,1-二苯基-2-三硝基苯肼(DPPH)自由基、羟自由基和2,2′-联氮-双-3-乙基苯并噻唑啉-6-磺酸(ABTS)自由基有一定的清除能力,其半抑制率含量(IC_(50))分别为2.30、0.01和1.00 mg·mL^(-1),对Fe^(3+)有一定还原能力。DPPH IC_(50)值和ABTS IC_(50)值与粗多糖和氮含量呈正相关,羟自由基IC_(50)值与粗多糖和氮含量呈负相关。本研究结果为黄蘑菇色素的开发利用提供了理论依据。