红花尔基不同龄级天然樟子松外生菌根真菌群落结构特征

红花尔基被誉为樟子松Pinus sylvestris var. mongolica的故乡,樟子松是我国北方主要的造林树种之一,也是典型的菌根依赖型树种。研究樟子松根围外生菌根真菌（ectomycorrhizal fungi,ECMF）群落结构特征对红花尔基沙地樟子松生态系统的保护具有重要意义。本研究以红花尔基天然樟子松根围土壤为研究对象,采用Illumina MiSeq测序技术研究了4个龄级（<10、11-20、21-30和31-40年,分别称作Ⅰ、Ⅱ、Ⅲ和Ⅳ龄级）天然樟子松根围土壤ECMF群落结构特征。结果表明：（1）测序共获得87 516条ECMF序列,划分为177个OTUs（operational taxonomic units,OTU）,隶属于2个门,4个纲,12个目,26个科,43个属。（2）Ⅳ龄级中Ace丰富度指数和Shannon-Wiener多样性指数均显著高于Ⅰ龄级（P<0.05）。（3）不同龄级根围ECMF优势属占比不同,Ⅰ龄级中占比最大的为Inocybe（42.55%）;Ⅱ和Ⅲ龄级中占比最大的均为Tricholoma,分别为31.64%和27.69%;Ⅳ龄级中占比最大的为Cortinarius（28.80%）。（4）冗余分析表明,土壤pH值对ECMF的影响程度最大,其次为速效钾和碱解氮;同时,不同理化因子对群落中优势属的影响也存在差异。     

基质辅助激光解析电离飞行时间质谱在医学真菌领域的应用进展

基质辅助激光解析电离飞行时间质谱（matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF MS）是近年来新兴的微生物检测技术,通过核糖体蛋白分析实现对真菌快速、准确鉴定。本文针对MALDI-TOF MS用于致病真菌鉴定、分类、体外抗真菌药物敏感性检测以及临床微生物样本直接检测等方面作一综述。     

兰科药用植物手参种子的真菌共生萌发

手参Gymnadenia conopsea是一种地生型兰科植物,是我国的传统中药,同时也是蒙药、藏药常用药,在西藏地区亦作食用。现代药理学研究表明手参具有非常好的药理活性,然而当前手参还不能进行人工栽培,市场需求完全依赖野生资源,因此资源短缺仍然是制约该种药用植物进一步开发利用的瓶颈。兰科植物种子萌发及早期的幼苗生长均需真菌参与,基于此,本文从手参根系中进行了真菌分离,并获得一株菌株,分子鉴定为角担菌属Ceratobasidium GS2。将GS2菌株与手参种子进行共培养,可明显促进手参种子的原球茎的形成并最终分化成幼苗。手参种子共生萌发的成功对于实现手参的种质保育、人工栽培和野生居群的生态恢复均具有重要的意义。     

AM真菌摩西管柄囊霉对干旱胁迫下枳抗氧化酶基因表达的影响

测定分析了接种丛枝菌根（AM）真菌摩西管柄囊霉Funneliformis mosseae对正常供水与干旱处理的盆栽枳Poncirus trifoliata实生苗生长、活性氧代谢及抗氧化酶基因表达量的影响。结果表明,7周干旱处理显著降低了根系菌根侵染率。接种摩西管柄囊霉显著促进了干旱处理的枳植株生长,增加了根系体积和叶片相对含水量,显著降低了叶片脯氨酸含量,同时也上调了干旱处理的枳叶片精氨酸脱羧酶基因（PtADC1和PtADC2）和超氧化物歧化酶基因（PtFe-SOD和PtMn-SOD）、过氧化物酶基因（PtPOD）和过氧化氢酶基因（PtCAT1）的表达,因而维持了一个相对更低的活性氧水平（如过氧化氢）,有利于增强植株的抗旱性。     

西双版纳5种附生兰非菌根内生真菌多样性研究

从西双版纳纳板河流域国家级自然保护区的盆距兰Gastrochilus、隔距兰Cleisostoma、毛兰Eria、贝母兰Coelogyne和万代兰Vanda的根、茎、叶分离得到172株非菌根内生真菌,根据ITS1-5.8S-ITS2 rDNA系统发育分析鉴定为盘菌亚门的4纲11目29属,其中炭角菌目、肉座菌目和格孢腔菌目真菌的相对频率最高;Xylaria、Fusarium和Phoma为主要优势属,相对频率分别为23.3%、14.5%和10.5%,它们的组织专化性不同,其中Xylaria在根、茎和叶均有分布,而Fusarium和Phoma都分离自根部。不同组织的内生真菌多样性和种类组成差异明显,根、茎和叶的内生真菌Shannon多样性指数分别为2.4083、1.0312和0.6557,表明根部内生真菌多样性显著高于茎、叶内生真菌多样性,其中根的主要优势内生真菌类群有Fusarium、Xylaria和Phoma,茎的主要优势内生真菌类群有Xylaria、Pestalotiopsis和Colletotrichum,叶的主要优势内生真菌类群有Colletotrichum、Pallidocercospora、Pantospora和Phyllosticta。从附生兰种类来看,盆距兰、隔距兰、毛兰、贝母兰和万代兰内生真菌的Shannon多样性指数分别为2.2689、2.2635、2.0115、1.9197、1.7139,表明盆距兰和隔距兰内生真菌多样性高于毛兰、贝母兰和万代兰。     

毛筒壳科真菌次级代谢产物生物活性的评价

毛筒壳科Tubeufiaceae真菌具有产新结构、新活性次级代谢产物的潜力,目前对该科真菌次级代谢产物的研究较少。为了寻找具有生物活性的新化合物,有必要对毛筒壳科真菌次级代谢产物及其活性进行系统深入的研究。本文采用平板对峙法、生长速率法和MTT法,分别测定已分离得到的19株该科真菌活体菌株抑菌活性、发酵物抑菌活性以及发酵物粗提物对不同人体肿瘤细胞株增殖的抑制作用。通过平板对峙法,试验共筛选获得13株活性菌株,其中,红棕毛筒腔菌菌株Tubeufia rubra PF02-2对7种植物病原真菌有明显的抑菌效果,抑制率均高于60%且抑菌谱广。采用生长速率法,发现红棕毛筒腔菌菌株PF02-2经液体发酵后,发酵液对其中4种植物病原真菌仍有一定的抑制作用,且菌丝体部分的乙酸乙酯提取物对马铃薯早疫病病菌Alternaria solani（ZYB）的抑制效果最好。通过MTT法,发现发酵物粗提物对3种肿瘤细胞均具有不同程度的细胞毒活性,其中在300μg/mL时,剑叶莎毛筒腔菌菌株Tubeufia machaerinae ML03-2发酵液部分的乙酸乙酯提取物对人宫颈癌细胞株HeLa和人前列腺癌细胞株PC-3的抑制率（%）分别达到了98.92±0.15和97.86±0.18,在400μg/mL时,对人肝癌细胞株HEPG2的抑制率（%）达到了98.88±0.04;在500μg/mL时,明孢新旋卷孢菌菌株Neohelicosporium hyalosporum ML05-1菌丝体部分的乙酸乙酯提取物对人宫颈癌细胞株HeLa细胞的抑制率（%）为98.32±0.02,在600μg/mL时,对人肝癌细胞株HEPG2的抑制率（%）达到了97.62±0.20,在300μg/mL时,对人前列腺癌细胞株PC-3的抑制率（%）达到了98.91±0.02。该研究结果为开发利用毛筒壳科真菌提供了科学依据。     

青岛胶州湾四种类型湿地AM真菌多样性

对青岛胶州湾4种类型湿地（盐田、滩涂、湖泊和河口）中芦苇Phragmites communis、香蒲Typha orientalis和碱蓬Suaeda glauca 根围土壤中丛枝菌根（arbuscular mycorrhizal,AM）真菌进行孢子分离与鉴定,分析湿地生态系统中植物根围AM真菌群落特征。共分离到AM真菌5属10种,包括斗管囊霉属Funneliformis 2种、无梗囊霉属Acaulospora 3种、近明球囊霉属Claroideoglomus 2种、球囊霉属Glomus 1种、巨孢囊霉属Gigaspora 2种,其中,斗管囊霉属Funneliformis及地斗管囊霉Funneliformis geosporum的分离频度和相对多度最高,分别为湿地中AM真菌优势属和优势种。滩涂和河口湿地中植物AM真菌侵染率显著高于湖泊和盐田湿地植物,AM真菌孢子密度以滩涂湿地最高（572个/20mL）,湖泊湿地最低（220个/20mL）;滩涂湿地的种丰度和Shannon-Wiener指数最高,分别为3.8和1.2。从植物种类来看,AM真菌侵染率总体呈现出香蒲>碱蓬>芦苇,AM真菌孢子密度以香蒲最高,芦苇最低,植物种类对AM真菌种丰度和Shannon-Wiener指数影响不显著（P>0.05）。二因素方差分析和典型RDA相关分析表明,寄主植物种类对AM真菌孢子密度有一定影响,但湿地类型对AM真菌多样性的影响更为显著（P<0.05）,胶州湾湿地土壤因子Ca ²⁺、速效P含量与AM真菌孢子密度、物种丰度和多样性指数显著负相关,而速效K、Na ⁺、pH与其显著正相关。结果表明植物种类主要影响AM真菌孢子密度,AM真菌多样性受植物种类和湿地类型综合影响,滩涂湿地较丰富的AM真菌多样性可能与该湿地较良好的理化性质有关。     

植菌昆虫与其共生真菌协同进化分子机制

昆虫菌业（fungiculture）是一种类似于人类种植业的昆虫种植体系,包括种植、耕作、收获和营养依赖4个过程,可分为高级的社会性昆虫如切叶蚂蚁、白蚁等和低级的非社会性昆虫如食菌小蠹虫、卷叶象甲、蜥蜴甲虫、树蜂等,它们均能种植并取食真菌。近年来随着组学及微生物组技术的发展,植菌昆虫与其共生真菌协同进化的分子机制研究方面取得了重要进展。系统发育分析阐明了植菌昆虫的起源与进化历程,并显示出与共生真菌系统发育的一致性;共生真菌细胞核数量也从双核增加到最多17个核,而染色体倍型也从单倍体增加为二倍体甚至多倍体;组学分析则揭示了植菌昆虫与其共生真菌在精氨酸、碳水化合物、木质素及几丁质合成或降解等方面显示出了高度的协同进化。本文系统综述了植菌昆虫及其共生真菌的系统进化、核进化及基因组进化进展,并探讨这种协同进化机制的生物学意义。     

Unravelling the interactions of the environmental host Acanthamoeba castellanii with fungi through the recognition by mannose‐binding proteins

Free‐living amoebae (FLAs) are major reservoirs for a variety of bacteria, viruses, and fungi. The most studied mycophagic FLA, Acanthamoeba castellanii (Ac), is a potential environmental host for endemic fungal pathogens such as Cryptococcus spp., Histoplasma capsulatum, Blastomyces dermatitides, and Sporothrix schenckii. However, the mechanisms involved in this interaction are poorly understood. The aim of this work was to characterize the molecular instances that enable Ac to interact with and ingest fungal pathogens, a process that could lead to selection and maintenance of possible virulence factors. The interaction of Ac with a variety of fungal pathogens was analysed in a multifactorial evaluation that included the role of multiplicity of infection over time. Fungal binding to Ac surface by living image consisted of a quick process, and fungal initial extrusion (vomocytosis) was detected from 15 to 80 min depending on the organism. When these fungi were cocultured with the amoeba, only Candida albicans and Cryptococcus neoformans were able to grow, whereas Paracoccidioides brasiliensis and Sporothrix brasiliensis displayed unchanged viability. Yeasts of H. capsulatum and Saccharomyces cerevisiae were rapidly killed by Ac; however, some cells remained viable after 48 hr. To evaluate changes in fungal virulence upon cocultivation with Ac, recovered yeasts were used to infect Galleria mellonella, and in all instances, they killed the larvae faster than control yeasts. Surface biotinylated extracts of Ac exhibited intense fungal binding by FACS and fluorescence microscopy. Binding was also intense to mannose, and mass spectrometry identified Ac proteins with affinity to fungal surfaces including two putative transmembrane mannose‐binding proteins (MBP, L8WXW7 and MBP1, Q6J288). Consistent with interactions with such mannose‐binding proteins, Ac–fungi interactions were inhibited by mannose. These MBPs may be involved in fungal recognition by amoeba and promotes interactions that allow the emergence and maintenance of fungal virulence for animals.     

Soil microbiome mediates positive plant diversity‐productivity relationships in late successional grassland species

Which processes drive the productivity benefits of biodiversity remain a critical, but unanswered question in ecology. We tested whether the soil microbiome mediates the diversity‐productivity relationships among late successional plant species. We found that productivity increased with plant richness in diverse soil communities, but not with low‐diversity mixtures of arbuscular mycorrhizal fungi or in pasteurised soils. Diversity‐interaction modelling revealed that pairwise interactions among species best explained the positive diversity‐productivity relationships, and that transgressive overyielding resulting from positive complementarity was only observed with the late successional soil microbiome, which was both the most diverse and exhibited the strongest community differentiation among plant species. We found evidence that both dilution/suppression from host‐specific pathogens and microbiome‐mediated resource partitioning contributed to positive diversity‐productivity relationships and overyielding. Our results suggest that re‐establishment of a diverse, late successional soil microbiome may be critical to the restoration of the functional benefits of plant diversity following anthropogenic disturbance.