大球盖菇原生质体再生及单核化特性的研究*

大球盖菇原生质体再生条件及单核化特性结果。原生质体再生速度极快,涂布平板３ｄ后肉眼即可见明显的再生菌落形成,在ＰＧＰＭ再生培养基上再生率为０．９７～２．０％,渗稳剂种类对再生率无明显影响,但可影响再生菌落形态,液体预培养１～２ｄ,再生率明显下降;大球盖菇原生质体单核化率高达７７．６％,且再生双核体和再生单核体在形成再生菌落时无时间差,其生长速度亦无快慢之分,液体预培养可显著减少单核化率,再生单核体中存在亲本两种交配型,但二者的比率不为１。     

基于F2群体的香菇遗传图谱构建及其在QTL定位中的应用

以171个F2双核体菌株为作图群体,通过相互配对的2个单核体的基因型推断双核体基因型,构建了第一张基于双核体群体的香菇遗传图谱。该图谱包含分布于15个连锁群的459个标记,覆盖长度为989.7cM,平均标记间隔为2.2cM。此外,以此双核体群体作为表型分离群体,定位了6个与香菇双核体菌丝生长速度相关的QTLs,位于5个连锁群上。采用全同胞单核体随机交配策略,易于构建相对大的双核体群体,用于连锁图构建和QTL定位。研究表明,在食用菌连锁图谱构建及QTL定位研究中,利用F2群体,可能为提高遗传作图效率,解决作图群体与表型分离群体间不一致问题提供新的途径。     

基于等位基因InDel快速检测黑木耳原生质体单核体

以黑木耳 Auricularia auricula-judae栽培菌株Au916三磷酸甘油醛脱氢酶（GAPDH）两个等位基因中的InDel为基础,以原生质体再生菌株为研究材料,建立了基于InDel的黑木耳原生质体单核体检测技术,并采用荧光显微镜检和配对试验对此方法进行了验证。应用InDel能清晰地区分黑木耳双核体和两种原生质体单核体,且不同的单核体之间可以进行交配,其杂交双核体能正常出耳,结果表明基于等位基因InDel标记可以快速准确地检测黑木耳原生质体单核体。在大规模测序获得真菌基因组信息的基础上,InDel标记在原生质体单核体鉴定中的应用将更加普遍。     

Development of SCAR Markers to Determine the Mating Types of Lepista nuda Protoplast Monokaryons

Lepista nuda (Bull. ex Fr.) Cooke belongs to Tricholomataceae and is an edible fungus with both economic and medical value. Mycelia were isolated from the fruiting bodies of L. nuda and were used to prepare the protoplast monokaryons. One hundred and fifteen monokaryons were obtained and their mating types were determined using somatic incompatibility tests. Protoplast monokaryons segregated into either the A₁B₁ or the A₂B₂ mating types. Inter-simple sequence repeats and sequence-related amplified polymorphism fingerprinting were used to analyse the mating types of these protoplast monokaryons and 16 sequence-characterised amplified region primers were developed to efficiently differentiate between the monokaryon mating types. Multiplex PCR analyses were also established. The data presented here outline a method for the precise and rapid identification of protoplast monokaryon mating types, which has the promise to shorten the period required for conventional crossbreeding.     

金针菇基因组测序及萜类合成关键基因分析

金针菇是我国一种重要的食药用真菌,具有较高的营养和药用价值,尤其是从金针菇中分离得到了多种麦角甾醇、倍半萜等萜类,具有抗菌、抗肿瘤和降血脂等功效。以往关于金针菇的研究多集中在营养成分、栽培和市场开发等方面,而很少报道有关生物活性成分的合成和代谢研究。从福建省金针菇主要栽培品种中获得原生质体单核化L11菌株,我们进行了金针菇基因组测序与初步分析,并利用生物信息学手段,研究了其主要的生物活性成分——萜类合成途径中的关键基因。结果显示,金针菇单孢全基因组序列长度为34.75Mb。通过分析萜类合成途径中的相关基因,确定了4个萜类合成关键基因的基因结构,并且表明其蛋白结构具有稳定性,该类基因还含有多个信号肽和跨膜结构。金针菇基因组测序的完成,为下一步功能基因的筛选、分析等研究工作奠定了基础。     

Effects of growth temperatures on plating efficiencies and stabilities of heterokaryons of Candida albicans

Heterokaryons (hets) of Candida albicans constructed by fusing protoplasts of complementing auxotrophs produce heterogeneous clones on minimal medium consisting of (i) a minority of slow-growing hets, (ii) a preponderance of non-growing, parental-type auxotrophic monokaryons, and (iii) some prototrophic monokaryons bearing hybrid nuclei. Hets grown at a given temperature within the range 25° C to 41° C replate with higher efficiencies at any lower temperature and exhibit progressively declining plating efficiencies as plate temperatures increase beyond that at which they were initially grown. Neither auxotrophic nor prototrophic monokaryons show such responses. Growth of colonies produced by hets, wild-type strains or prototrophic hybrid monokaryons is stimulated by temperatures in the order, 37° C > 30° C > 41° C > 25° C. However, the proportion of hets to auxotrophic monokaryons within individual net clones increases directly from 25° C to 41° C. Though this pattern obtains whether colonies are compared at equivalent sizes or ages, het frequencies decline as colonies age at all temperatures. Appearance of hybrid monokaryons within het clones is unaffected by growth temperature. The relationships of temperatures to plating efficiencies and stabilities of hets are independent of the natures of their complementing auxotrophies or the wild-type backgrounds of their nuclear components and are, therefore, functions of heterokaryosis per se. Modifications of these relationships by selective metabolic antagonists or by growth of hets on different preand post-plating carbon sources indicate that they reflect temperature-dependent properties of mitochondria which are peculiar to hets.     

Monokariotic fruiting body and clamp cell formation in Mycoleptodonoides aitchisonii (Bunaharitake)

The ability to produce monokaryotic fruiting bodies and clamp cells in culture was examined in monokaryotic strain isolated from several dikaryotic parental strains of the edible mushroom, Mycoleptodonoides aitchisonii (Bunaharitake). We describe a single dikaryotic M. aitchisonii strain, TUFC50005, and 20 monokaryons derived from it, which exhibited a wide spectrum of monokaryotic fruiting types. Most strains formed primordia, or young fruiting body-like structures, but only one of the monokaryons, strain TUFC50005-4, formed a fruiting body, even though it had only one nucleus and produced only two spores after meiosis. We demonstrated that dikariotization was not required for clamp cell formation, fruiting body formation, or meiosis, in this mushroom.     

“食用菌产量和品质形成的分子机理及调控”项目简介——食用菌产业发展与技术创新的科学基础

食用菌以子实体为收获物,是优质蛋白的重要来源,我国重要的食物安全战略产业,已经成为我国粮、菜、果、油之后的第五大产值农作物,2013年产量3 169.68万吨（中国食用菌协会统计）,直接产值1 707亿元,间接产值6 828亿元,从业人口逾2 000万。我国食用菌产量已占全球总产量的75%以上。但是,与食用菌产业相关的基础科学研究严重匮乏,对产量和品质形成的关键科学问题研究的空白,严重制约着我国食用菌育种和栽培等产业技术的创新,导致单产低、质量差,产业链难以延长,产业效益持续下降,产业升级艰难。科学基础研究的缺乏,已经成为食用菌产业发展的瓶颈。2014年立项的国家重点基础研究发展计划（973）项目“食用菌产量和品质形成的分子机理及调控”（2014CB138300）,紧紧抓住困扰我国食用菌产业发展的科学问题,开展食用菌高效利用木质纤维素的分子机制、食用菌子实体形成发育的调控机理、食用菌温度响应的分子机制、食用菌活性物质及其合成代谢的分子基础和食用菌优异种质性状形成的遗传基础研究,拟阐释食用菌子实体形成的营养利用与遗传调控机制、食用菌抗逆性的温度响应机制和食用菌活性物质的合成代谢调控机理。构建食用菌科学理论体系,为食用菌产业关键技术创新奠定科学基础,为优质高产提供理论指导,为延长产业链,提高产业效益提供科学路径。     

The constitution of incompatibility factor and mating characteristics of spore isolates in a bipolar mushroom, <Emphasis Type="Italic">Pholiota nameko</Emphasis>

Pholiota nameko is a wood-rotting edible mushroom that carries a bipolar A incompatibility factor gene. The linkage analysis of the multiple allelomorphic A factor gene demonstrated that sexual reproduction produced a monospore isolate carrying a new A factor gene in addition to two parental mating types of isolates. However, 10%–30% of the modified monospore isolates could not produce a dikaryon with both of the parental monokaryons by crossing. It is concluded that the bipolar A incompatibility factor gene of P. nameko is constituted of two functional subunits, Aα and Aβ, which might be successively located beside each other with an apparent genetic distance of 0.3 centi-Morgan between them on the same chromosome. Further, some monospore isolates that did not conjugate with both parental monokaryons could produce dikaryons with different monokaryotic stocks with either one of the parental mating types. This result suggests that the crossing capability of these isolates were essentially those for one of the mating types of the parental monokaryons, but that their function for mating activity was made partially by unequal crossing-over in the process of sexual recombination. Received: May 1, 2001 / Accepted: December 5, 2001     

食用蕈菌硒多糖研究进展

食用蕈菌硒多糖是无机硒与蕈菌多糖结合而成的有机硒化合物,兼有二者的活性。该文综述了食用蕈菌硒多糖的组成分析、提取、分离纯化以及生理功能、应用现状等方面的研究进展。     

Mushroom diversity in sustainable shade tea forest and the effect of fire damage

A survey of the biodiversity of wild macrofungi, including edible species yields, was carried out from 1 May to 30 September 2007 at four different forest types (in mainly Miang tea forest). The plots 100 m², comprised a tea garden with a few planted canopy tree species (37.2% canopy cover), a cultivated sustainable tea forest (80.2% canopy cover), an abandoned sustainable tea forest (89.8% canopy cover), and an abandoned sustainable tea forest that had suffered fire damage (72.9% canopy cover). All visible mushrooms were collected during weekly forays. Macro-characters of the fungi were annotated and photographed by digital camera and the fungi were identified to genera and morphospecies (e.g. Agaricus sp.1, Agaricus sp.2). The fresh weight of wild edible mushrooms produced in the plots was also recorded during this period. The biodiversity of macrofungi in abandoned sustainable tea forest was highest with 115 species in 47 genera, followed by cultivated sustainable tea forest with 85 species in 42 genera and fire damaged abandoned sustainable tea forest with 48 species in 25 genera, while only 19 species belonging to 15 genera were found in the tea garden. Twenty-one species belonging to nine genera with a total of 60.9 kg of edible fungi were collected throughout this study. The fresh weight of edible mushrooms was recorded with the highest fresh weight (25.3 kg) collected from the abandoned sustainable tea forest, followed by 18.2 kg in the cultivated sustainable tea forest, while in the fire damaged sustainable tea forest 16.4 kg were collected and only 1 kg was collected in the tea garden. Abandoned sustainable tea forest contained a greater shade tree biodiversity and higher canopy cover than other plots. This area has a generally higher humidity, a greater ground litter cover, a greater number of microhabitats, a greater number of trees that can form mycorrhizal associations and this probably accounts for the higher diversity of macrofungi and production of edible mushrooms. The fire damaged sustainable tea forest had lower mushroom diversity and edible mushroom production possibly due to the loss of litter and lower tree diversity. The tea garden supported a poor diversity of mushrooms and almost no edible mushrooms. Shade tea forest (Miang tea forest) is a good method to produce tea in a sustainable way as it maintains diversity of mushrooms and other organisms and could be developed as an alternative to shade coffee.     

云南省常见野生食用菌的氨基酸含量

采用国家标准方法对云南41种常见野生食用菌鲜菌中17种氨基酸进行了定量,结果表明这些食用菌中氨基酸种类齐全、含量丰富,其中玉蕈离褶伞、灰褐牛肝菌、球形鸡枞菌和条纹鸡枞菌等总氨基酸含量较高。     

Amino Acids Contents of Common Wild Edible Mushrooms in Yunnan Province

Edible mushrooms are well known for their value of nutrition. We analysed the contents of 17 amino acids in 41 species of common wild edible mushrooms in Yunnan Province. The results indicate that the types of amino acids are rich and the contents of amino acids are high. The total amino acids contents of Lyophyllum shimeji, Boletus griseus, Termitomyces bulborhizeus, and T.striatus are rather higher than others.     

我国食用菌遗传学的发展及展望

食用菌遗传学是食用菌学科体系的重要分支之一,40年来我国食用菌遗传学研究紧密围绕为育种服务、最新分子生物学技术的应用和生产实践中的科学问题的解决等主题开展了众多科学活动.为了促进食用菌遗传学研究的系统性和全面性,本文梳理出9个方面的研究主题,主要包括食用菌种质资源调查和地方品种研究、食用菌农艺性状控制基因定位和分子辅助...     

Production of edible mushrooms in forests: trends in development of a mycosilviculture

Developing the production of ectomycorrhizal (ECM) mushrooms in forest has become a challenge. Only a few ECM species are currently cultivable. Controlled mycorrhization practices offer promising advance to produce currently uncultivable ECM mushrooms. The persistence of the production of edible species, either cultivated or wild, depends on both the tree and the ecological environment (fungal communities, climate, soil, tree development). Developing adapted forest management practices appears to be means to improve production of edible ECM mushrooms. This review summarises current knowledge on the development of a science-based mycosilviculture for the production of edible ECM mushrooms.     

Explaining Dioscorides' "double difference": why are some mushrooms poisonous, and do they signal their unprofitability?

The adaptive significance of toxins in mushrooms has received very little consideration, although it is clear that poisons have appeared (and/or disappeared) many times in mushrooms' evolutionary history. One possibility is that poisons have evolved in some mushroom species to deter their consumption by would-be fungivores before spore dispersal. If this is so, then one might expect poisonous mushrooms to signal their unprofitability in some way. In this study, we have conducted the first formal analysis of the ecological and morphological traits associated with edible and poisonous mushrooms in North America and Europe. Poisonous mushrooms do not tend to be more colorful or aggregated than edible mushrooms, but they are more likely to exhibit distinctive odors even when phylogenetic relationships are accounted for. This raises the intriguing possibility that some poisonous species of mushrooms have evolved warning odors (and perhaps tastes) to enhance avoidance learning by fungivores.     

Identification of Degenerate Nuclei and Development of a SCAR Marker for Flammulina velutipes

Flammulina velutipes is one of the major edible mushrooms in the world. Recently, abnormalities that have a negative impact on crop production have been reported in this mushroom. These symptoms include slow vegetative growth, a compact mycelial mat, and few or even no fruiting bodies. The morphologies and fruiting capabilities of monokaryons of wild-type and degenerate strains that arose through arthrospore formation were investigated through test crossing. Only one monokaryotic group of the degenerate strains and its hybrid strains showed abnormal phenotypes. Because the monokaryotic arthrospore has the same nucleus as the parent strain, these results indicated that only one aberrant nucleus of the two nuclei in the degenerate strain was responsible for the degeneracy. A sequence-characterized amplified region marker that is linked to the degenerate monokaryon was identified based on a polymorphic sequence that was generated using random primers. Comparative analyses revealed the presence of a degenerate-specific genomic region in a telomere, which arose via the transfer of a genomic fragment harboring a putative helicase gene. Our findings have narrowed down the potential molecular targets responsible for this phenotype for future studies and have provided a marker for the detection of degenerate strains.     

香菇135菌株特异SCAR标记在其原生质体单核中的分布

以香菇保藏菌种庆元135以及庆元出菇的135新鲜子实体组织分离得到的菌丝为材料制备原生质体,分别获得58和83个原生质体单核体;经交配型鉴定后,用已获得的135特异SCAR引物对单核进行PCR扩增,统计SCAR条带的分布。结果显示：原生质体单核体分为A1B1和A2B2两种交配型,而特异条带仅存在于后者中,证明135特异SCAR标记是稳定遗传的,也为鉴定以带标记核为亲本之一的杂交后代提供了依据,这就进一步拓宽了分子标记应用于菌株鉴定的适用范围。     

云南普洱地区野生食药用菌及其持续利用

2002—2009年夏,对云南省普洱市澜沧县、景东县及其周边地区野生食用菌的物种多样性、用途或特性以及濒危等级进行了调查研究,共采集、鉴定标本593份,并对采于该区的214份馆藏标本进行了统计,野生食用菌共计35科61属134种(含变种、变型)。同时对该区市场上出售的野生贸易类群也进行了调查统计,有28科39属95种(含变种、变型)。迄今普洱共计有野生食用菌38科67属155种(含变种、变型),其中多孔菌科(Polyporaceae)所含种数最多,有17种,占总种数的10.97%;红菇科(Russulaceae)和牛肝菌科(Bo-letaceae)皆为16种,共占总种数的20.65%,灵芝菌科(Ganodermataceae)有11种,占总种数7.1%。在这些类群当中,木耳属(Auricularia)、牛肝菌属(Boletus)、乳菇属(Lactarius)、枝瑚菌属(Ramaria)、红菇属(Russula)、蚁巢菌属(Termitomyces)和革菌属(Thelephora)等属的42种真菌为普洱市场上最常见的野生贸易类群,约占云南优势野生贸易真菌总种数的52.5%。近年来,随着人们对野生食用菌消费需求的日益增长,导致其商业化采收规模急剧扩大,造成生存环境的破坏甚至丧失,从而使普洱野生食用菌,尤其是野生贸易类群的产量呈逐年下降趋势。因此,亟待采取行之有效的措施保护和管理这些珍贵的自然资源,以期达到持续利用之目的。