松口蘑人工菌塘诱导形成的初步研究

本研究通过土壤筛选找到适合松口蘑菌丝体生长的土壤,并在其土壤中添加适当浓度的橄榄油(0.5%-2.0%),有效地促进了菌丝体在土壤中的生长,菌丝生长状况酷似野外的松口蘑菌塘,且随着添加浓度的增加,菌丝体生物量呈明显的正相关。当土壤中橄榄油浓度为0.5%和1.0%时,松口蘑共生植物赤松苗的生长不但没有受到抑制,而且还能与菌丝体在这种“菌塘”中形成菌根。本研究成功地建立了中国松口蘑人工菌塘诱导体系,同时也为其人工子实体的诱导研究提供了必要的前提条件。     

伞菌DNA简易序列分析引出的思考

目的:伞菌物种以子实体形态特征为分类依据,研究以菌丝体代替子实体进行种质资源鉴定的遗传证据。方法:以常见的食用伞菌香菇、平菇子实体的不同部位组织及其分离菌丝体为供试材料,制备了12个随机引物介导的RAPD-PCR指纹,把DNA指纹图谱转化为简易的序列数据,经生物信息处理软件比较分析。结果:香菇不同菌株子实体DNA相似性系数在0.886～0.986之间,平菇不同菌株子实体DNA相似系数在0.779～0.976之间。对于供试的单个子实体而言,香菇和平菇子实体的菌盖、菌褶、菌柄及其组织分离菌丝体,与以此为菌种栽培得到的子实体相比较,所获得的有限DNA指纹图谱全部相同。结论:揭示了香菇和平菇不同菌株的遗传多样性,初步反映了伞菌不同发育阶段在分子水平上的遗传变异和亲缘关系,争论了以菌丝体代替子实体使用RAPD手段进行种质资源鉴定与系统发育分析引出的真菌遗传问题。     

应用蛋白酶抑制剂和诱导物质提高食用菌产量

菌蕈是指担子菌的子实体,如蘑菇、香菇、金针菇、平菇、玉蕈、松口蘑以及灵芝等,食用或药用菌蕈.近年来随着人工栽培食用菌蕈的发展,其产量飞跃增加,在农产品中占有重要的地位.菌蕈是由双核化营养菌丝体在温度和光的刺激下形成子实体原基,利用营养菌丝和培养基中的养分发育成为子实体的,关于菌蕈的生化研究进展相当迟缓,因此营养菌丝体向原基转变,进而长成子实体的机理几乎不了解.搞清子实体形成机理,将     

抗真菌抗菌素414的研究I．球孢玫瑰紫链霉菌(Streptomycesgloboroseoviolaceus n. sp.)的分类学研究

从我国江西南昌土壤中分离到一株抗真菌抗菌素414的产生菌,具有松螺旋形的孢子丝,气生菌丝体玫瑰粉红色,基内菌丝体紫红褐色和无可溶性色素等特征。根据形态和在各种培养基上的培养特性以及其他生理特性等方面的研究,该菌株系属于球孢链霉菌类群,但不同于所有已知的种,定名为球孢玫瑰紫链霉菌(Streptomyces globoroseoviolaceus n. sp.)。     

大白口蘑分离菌株的DNA鉴定

以松口蘑Tricholoma matsutake子实体为外类群,对大白口蘑T.giganteum野生子实体及其组织分离菌丝进行ITS序列测序,通过DNAStar软件进行比较分析。结果表明大白口蘑ITS序列长度为589bp,松口蘑ITS序列长度为601bp,ITS1和ITS2呈现不同程度的种间多态性;ITS序列测定证实了大白口蘑野生子实体及其组织分离菌丝的同质性,并且ITS区序列在大白口蘑种内不同菌株间的变异程度很小,表明使用通用引物ITS4和ITS5,通过PCR扩增测序即可用于大白口蘑的种质鉴定。     

大白口蘑分离菌株的DNA鉴定

以松口蘑Tricholoma matsutake子实体为外类群,对大白口蘑T.giganteum野生子实体及其组织分离菌丝进行ITS序列测序,通过DNAStar软件进行比较分析。结果表明大白口蘑ITS序列长度为589bp,松口蘑ITS序列长度为601bp,ITS1和ITS2呈现不同程度的种间多态性;ITS序列测定证实了大白口蘑野生子实体及其组织分离菌丝的同质性,并且ITS区序列在大白口蘑种内不同菌株间的变异程度很小,表明使用通用引物ITS4和ITS5,通过PCR扩增测序即可用于大白口蘑的种质鉴定。     

大白口蘑分离菌株的DNA鉴定

以松口蘑Tricholoma matsutake子实体为外类群,对大白口蘑T. giganteum 野生子实体及其组织分离菌丝进行ITS序列测序,通过DNAStar软件进行比较分析。结果表明大白口蘑ITS序列长度为589bp,松口蘑ITS序列长度为601bp,ITS1和ITS2呈现不同程度的种间多态性;ITS序列测定证实了大白口蘑野生子实体及其组织分离菌丝的同质性,并且ITS区序列在大白口蘑种内不同菌株间的变异程度很小,表明使用通用引物ITS4和ITS5,通过PCR扩增测序即可用于大白口蘑的种质鉴定。     

嗜热放线菌类群分类的研究V．小单孢菌属的分类鉴定

从我国各地的土壤、堆肥及粪肥中分离到一批嗜热小单孢菌,经52℃培养,其中’524菌株在固体培养基上有发育良好的基内菌丝体,在基内菌丝体上着生茄子形孢子,细胞壁组分II型,含内消旋二氨基庚二酸、甘氨酸。此菌株为罕见的嗜热菌株,区别于已报道的小单孢菌属中的种,故认为是个新种,命名为热茄孢小单孢菌(Micromonospora thermoauberginospora n.sp.)。     

珍稀共生食用菌松口蘑的人工驯化

松口蘑是与松树共生的一种外生菌根真菌 ,被国内外誉为“蘑菇之王” ,极其珍稀昂贵。松口蘑的研究内容涉及到与近缘种的进化关系 ,与松树共生的生态环境 ,菌丝体的分离 ,纯化 ,培养方法及其真伪鉴定 ,纯培养的菌根建成 ,以及松口蘑半人工栽培试验等方面。讨论了目前人工驯化栽培松口蘑存在的问题及可能的解决途径。     

云南干巴菌子实体内可培养微生物

【背景】微生物在菌根真菌的孢子萌发、菌丝体生长、菌根形成以及子实体发育等过程中起到一定作用。【目的】对采自云南省昆明市嵩明县和楚雄彝族自治州禄丰县的8个干巴菌子实体内的微生物进行分离培养鉴定,为后期研究微生物与干巴菌之间的相互作用奠定基础。【方法】采用传统平板分离法从干巴菌子实体内分离获得微生物群落,t检验分析不同地区采集的干巴菌子实体内微生物菌落总数的差异,16S r RNA基因和ITS序列进行系统发育树构建和微生物多样性分析。【结果】采自嵩明县和禄丰县的8个干巴菌子实体内共分离获得282株可培养的细菌,两个地区的细菌菌落总数无显著差异(P=0.22)。所有细菌分属2门12属15种。其中80%的细菌属于变形菌门,且以γ-变形菌为优势菌群,假单胞菌属(Pseudomonas)为优势菌属。其余20%的细菌属于拟杆菌门。从干巴菌子实体中分离获得114株真菌,两个地区的真菌菌落总数无显著差异(P=0.65)。所有真菌分属2门10属10种。其中62%的真菌属于子囊菌门(Ascomycota),并以分离自禄丰县干巴菌子实体内的Lophiostoma为优势属。38%的真菌属于担子菌门(Basidiomycota),并以Asterotremella为优势属。【结论】两个不同地区采集的干巴菌子实体内细菌和真菌在菌落总数上无显著差异。所有细菌都以γ-变形菌为优势菌群,假单胞菌属为优势菌属。嵩明干巴菌子实体内真菌以担子菌门为优势菌群,Asterotremella为优势属。而禄丰干巴菌子实体内真菌则以子囊菌门为优势菌群,Lophiostoma为优势属。     

利用松茸ITS特异性引物对松茸分离物进行鉴定

利用一对ITS通用引物(YIS4-ITS5)和一对松茸物ITS特异性引物(TMF-TMR)对来源于云南省不同地区的6个松茸(Tricholoma matsutake)子实体及其6株分离物,3个假松茸(Tricholoma bakamatsutake)子实体及其3株分离物、侧耳(Pleurotus astreatus)、金针菇(Flammulina velutipes)和双孢蘑菇(Agaricus bisporus)的子实体进行了PCR扩增,琼脂糖凝胶电泳分析,结果表明ITS4-ITS5能将所有的样品扩增,并得到600bp左右的DNA扩增条带,TMFTMR扩增时,只有松茸子实体及其对应分离物有扩增条带,DNA片段大小在500bp左右。进一步对松茸子实体(TG25)及其分离物(TM25112．2)进行WS序列测定表明两者的DNA同源性为100％,从而证明所分离到的6个菌株确为松茸的纯培养物。     

Traceability of Asian Matsutake, specialty mushrooms produced by the ectomycorrhizal basidiomycete Tricholoma matsutake, on the basis of retroelement-based DNA markers

The ectomycorrhizal basidiomycete Tricholoma matsutake produces commercially valuable fruit bodies, matsutake, in forests. Here we report a PCR system targeting retroelement integration sites to differentiate among individual Asian isolates of T. matsutake based on their geographical origins, such as Japan, the area of South Korea through North Korea, the northeastern provinces of China, and the area of the southwestern provinces of China through Bhutan. The overall misjudgment rate of the analytical system was approximately 5% based on 95 samples of T. matsutake examined including those from cultures and from commodities. We also provide evidence that T. matsutake isolates grown throughout the Far East, including the northeastern provinces of China, are closely related to each other while distinct from those in the area of the southwestern provinces of China through Bhutan. The method allows us to trace back geographical origins of Asian matsutake, thus contributing to food safety, appropriate tariffs, and proper price setting.     

四川省雅江松茸菌的分离与系统发育

对外生菌根真菌-松茸的纯培养条件进行了探讨,并从采集自四川省雅江县的松茸子实体中获得了10株松茸菌的纯培养物;分别以NS1和NS6 ,NS1和NS8,ITS4和ITS5为引物,对分离获得的松茸菌进行了18S r DNA PCR- RFL P和ITS PCR-RFL P分析,结果显示,用Alu I,H ae III,H inf I和Msp I四种限制性内切酶,这些松茸菌株的18S r DNA、ITS片段的酶切图谱完全相同;代表菌E7的ITS序列分析结果表明,本研究分离的松茸菌与Tricholoma matsutake的菌株在系统发育上高度同源,在分类上应属于同一个种     

松口蘑复合种形态学及生物地理学研究

松口蘑复合种含有松口蘑Tricholoma matsutake (也称松茸)、北欧口蘑T. nauseosum、喜栎口蘑T. zangii和美洲口蘑T. magnivelare等4个分类单元。在对来自中国、日本、法国、西班牙、瑞典、芬兰和美国的标本进行形态学研究的基础上,认为松口蘑、北欧口蘑、喜栎口蘑形态十分接近,它们的分类地位应该重新考虑。美洲口蘑和它们的差别也不大。对松口蘑复合种的寄主和地理分布也进行了总结,发现松口蘑和美洲口蘑的寄主范围较广,寄主专化性不强,而北欧口蘑和喜栎口蘑的寄主比较专一。在地理分布上,松口蘑和美洲口蘑形成亚洲—北美地理替代种的分布格局;松口蘑和北欧口蘑形成欧亚间断分布的分布格局;喜栎口蘑的地理分布范围完全包含在松口蘑的分布范围中。松口蘑复合种种类的分布区和它们各自寄主的分布不一定完全相同。在松口蘑复合种内,北欧口蘑和松口蘑已被认为是同一个种,而其余两个分类单元的分类地位值得进一步研究。     

Tricholoma matsutake in a natural Pinus densiflora forest: correspondence between above- and below-ground genets, association with multiple host trees and alteration of existing ectomycorrhizal communities

Tricholoma matsutake (matsutake) is an ectomycorrhizal (ECM) fungus that produces economically important mushrooms in Japan. Here, we use microsatellite markers to identify genets of matsutake sporocarps and below-ground ECM tips, as well as associated host genotypes of Pinus densiflora. We also studied ECM fungal community structure inside, beneath and outside the matsutake fairy rings, using morphological and internal transcribed spacer (ITS) polymorphism analysis. Based on sporocarp samples, one to four genets were found within each fairy ring, and no genetic differentiation among six sites was detected. Matsutake ECM tips were only found beneath fairy rings and corresponded with the genotypes of the above-ground sporocarps. We detected nine below-ground matsutake genets, all of which colonized multiple pine trees (three to seven trees per genet). The ECM fungal community beneath fairy rings was species-poor and significantly differed from those inside and outside the fairy rings. We conclude that matsutake genets occasionally establish from basidiospores and expand on the root systems of multiple host trees. Although matsutake mycelia suppress other ECM fungi during expansion, most of them may recover after the passage of the fairy rings.     

Artificial shiro formation of Tricholoma matsutake

One type of soil collected from Maoer-shan in Heilongjiang Province,China was selected to induce hyphal growth of Tricholoma matsutake by a soil screening experiment.It was confirmed that hyphal growth of all the tested T.matsutake isolates was significantly stimulated in soil by supplemented with 0.5%～2.0% olive oil.The aggregation of hyphae and soil resembled natural Shiro.The biomass of hyphae in the soil increases with increasing olive oil concentrations.Moreover,seedlings of Pinus densiflora grew well in the soil containing 0.5%～1.0% olive oil and were also successfully infected by T.matsutake isolate A in the soil containing 1.0% olive oil.This study established a culture system of artificial Shiro formation and also provided a premise for formulation of culture substratum for fruit body formation of T.matsutake.