线虫草科弯颈霉属研究进展

弯颈霉属Tolypocladium是W. Gams于1971年建立的新属,该属的特点是瓶梗基部呈球形或椭圆形膨大,瓶颈细长而弯曲,分生孢子单生或黏聚成小头状,截至目前国际真菌名录数据库中记录弯颈霉属真菌有43种。其模式种为膨大弯颈霉Tolypocladium inflatum W.Gams。弯颈霉属是一类重要虫生真菌,生态分布广,寄主种类多,弯颈霉属很多种被发现是虫草属无性型,与虫草属的关系十分密切,2014年一些国际著名真菌学家基于分子生物学的新分类系统将弯颈霉属归入到线虫草科Ophiocordicipitaceae。弯颈霉属能产生多种杀虫和抗菌的有效成分,如环孢菌素(Cyclosporin)、大团囊素(Ophiocordin,蛋白酶强抑制剂)、线肽素(efrapeptin)、四聚胺酸素(Ophiosetin)、多种杀虫剂等。文中综述了弯颈霉属真菌的分类、生活习性与生态分布等,介绍了其药理药效及有效化学成分等相关研究,并展望了弯颈霉属真菌的研究发展前景。     

林火对植物根围丛枝菌根真菌多样性的影响

林火是森林生态系统的一种主要干扰因子,以青岛市三标山林火迹地为研究对象,采集荆条(Vitex negundo)、胡枝子(Lespedeza bicolor)、花木蓝(Indigofera kirilowii)、青花椒(Zanthoxylum schinifolium)和野青茅(Deyeuxia arundinacea)5种优势植物根围土壤,研究不同林火强度对丛枝菌根(AM)真菌多样性的影响。结果表明,AM真菌侵染率和孢子密度随火灾强度的加强而降低;非过火区植物根围土壤中,分离鉴定出AM真菌3属11种,轻度过火区分离鉴定出AM真菌3属10种,中度过火区分离鉴定出AM真菌3属9种,重度过火区分离鉴定出AM真菌3属8种。过火区AM真菌种丰度低于非过火区。过火区和非过火区AM真菌的重要值和优势种不同,非过火区植物根围的优势种是地球囊霉(Glomus geosporum)、台湾球囊霉(G.taiwanensis)、分支巨孢囊霉(Gigaspora ramisporophora)、极大巨孢囊霉(Gi.gigantean)、福摩萨球囊霉(G.formosanum)、悬钩子球囊霉(G.rubiforme)、柯氏无梗囊霉(Acaulospora koskei)和松蜜无梗囊霉(A.thomii);轻度过火区植物根围的优势种是地球囊霉和台湾球囊霉;中度过火区的是台湾球囊霉和地球囊霉(野青茅除外);重度过火区植物根围的优势种是地球囊霉。不同强度的过火区对AM真菌群落组成有不同程度的影响。认为林火降低植物根围土壤中AM真菌多样性。     

金沙江干热河谷(元谋段)丛枝菌根真菌多样性研究*

从金沙江干热河谷(元谋段)75种植物的根际土壤中分离鉴定了44种丛枝菌根真菌, 分属无梗囊霉属Acaulospora、古孢霉属Archaeospora、内养囊霉属Entrophospora、巨孢囊霉属Gigaspora、球囊霉属Glomus和盾巨孢囊霉属Scutellospora, 其中,球囊霉属和无梗囊霉属为金沙江干热河谷中丛枝菌根真菌的优势属。齿状无梗囊霉A. denticulata、刺状无梗囊霉A. spinosa、瘤状无梗囊A. tuberculata,近明球囊霉Glomus claroideum、明球囊霉G. clarum、根内球囊霉G. intraradices、单孢球囊霉G. monosporum、弯丝球囊霉G. sinuosa是金沙江干热河谷(元谋段)的优势种。金沙江干热河谷土壤中丛枝菌根真菌的孢子密度为5~6400个/100g土壤,平均1504;每个根际土壤中丛枝菌根真菌的物种丰富度1~18种,平均9种。     

金沙江干热河谷(元谋段)丛枝菌根真菌多样性研究

从金沙江干热河谷(元谋段)75种植物的根际土壤中分离鉴定了44种丛枝菌根真菌, 分属无梗囊霉属Acaulospora、古孢霉属Archaeospora、内养囊霉属Entrophospora、巨孢囊霉属Gigaspora、球囊霉属Glomus和盾巨孢囊霉属Scutellospora, 其中,球囊霉属和无梗囊霉属为金沙江干热河谷中丛枝菌根真菌的优势属。齿状无梗囊霉A. denticulata、刺状无梗囊霉A. spinosa、瘤状无梗囊A. tuberculata,近明球囊霉Glomus claroideum、明球囊霉G. clarum、根内球囊霉G. intraradices、单孢球囊霉G. monosporum、弯丝球囊霉G. sinuosa是金沙江干热河谷(元谋段)的优势种。金沙江干热河谷土壤中丛枝菌根真菌的孢子密度为5~6400个/100g土壤,平均1504;每个根际土壤中丛枝菌根真菌的物种丰富度1~18种,平均9种。     

不同生境下川麦冬根围土壤丛枝菌根真菌多样性北大核心CSCD

采用高通量(Illumina Miseq)测序技术对栽培和野生2种生境下川麦冬根围的丛枝菌根(AM)真菌多样性和群落结构进行测定,并结合土壤理化因子进行相关性分析,以明确两种生境下川麦冬根围土壤AM真菌多样性和优势群落的分布特点,探讨AM真菌群落分布差异的驱动因子,为AM真菌应用于麦冬生产提供理论依据和技术支持。结果表明:(1)不同生境下川麦冬根围土壤中共鉴定出AM真菌3属10种,其中野生川麦冬根围土壤鉴定出的AM真菌3属7种,分别隶属于无梗囊霉属(Acaulospora)、多孢囊霉属(Diversispora)和球囊霉属(Glomus),而栽培环境下鉴定出AM真菌1属6种,隶属于球囊霉属。2个生境优势属均为球囊霉属。(2)不同生境下川麦冬根围AM真菌之间存在显著差异,野生生境下川麦冬根围土壤AM真菌多样性指数ACE和Shannon均显著高于人工栽培生境,而Simpson指数则相反。(3)相关性分析表明,AM真菌多样性指数及群落组成结构均与土壤理化因子存在相关性,其中全钾(TK)、全磷(TP)、全氮(TN)对AM真菌多样性指数和群落结构组成均存在显著影响。研究认为,不同生境下川麦冬根围AM真菌群落存在显著差异,球囊霉属为川麦冬互利共生的关键属,TK、TP、TN是不同生境川麦冬根围AM真菌群落差异的主要驱动因子。     

西藏高原针茅草地土壤因子对丛枝菌根真菌物种多样性的影响

在对西藏高原北部针茅草地根围土壤中的丛枝菌根(AM)真菌种类分离鉴定基础上,研究了藏北针茅草地的土壤质地、pH、有机质和有效磷含量对AM真菌孢子密度、分离频度、相对多度、重要值、物种多样性指数和均匀度的影响.结果表明: 针茅草地根围土壤中共分离鉴定出AM真菌3属15种,其中,球囊霉属9种、无梗囊霉属6种、盾巨孢囊霉属1种.球囊霉属和无梗囊霉属为藏北针茅草地AM真菌的优势属;近明球囊霉和光壁无梗囊霉为藏北高寒草原针茅属植物根围AM真菌的优势种.不同质地土壤中AM真菌孢子密度、分离频度、相对多度和重要值均表现出球囊霉属＞无梗囊霉属＞盾巨孢囊霉属的趋势;土壤pH值对AM真菌种群组成无明显影响,球囊霉属和无梗囊霉属真菌分离频度、相对多度和重要值随土壤pH升高而增加,盾巨孢囊霉属则呈现相反趋势;不同土壤有机质含量范围内,AM真菌孢子密度等各项指标均呈球囊霉属＞无梗囊霉属＞盾巨孢囊霉属,而AM真菌属的分布没有明显规律;土壤有效磷含量对AM真菌种丰度和孢子密度影响较小.研究区域内AM真菌物种多样性指数和均匀度随着土壤有效磷含量升高而增加.     

地生弯颈霉的培养与生长条件

从长白山北坡森林土壤中分离纯化得到1株地生弯颈霉(Tolypocladium geodes),对其培养及生长条件(如培养基、光照、温度、碳源、氮源和pH等)进行了试验。结果表明,地生弯颈霉在麦芽浸膏琼脂培养基(MEA)上菌落生长直径最大,马铃薯葡萄糖琼脂培养基(PDA)上产孢量最高;5~30℃温度范围内均能生长,25℃时生长和产孢量最佳;最适碳源和氮源分别为葡萄糖和柠檬酸铵;光暗交替条件下,最利于其生长和产生孢子;pH4~9范围内均能生长和产孢,pH=8时菌落直径最大,pH=6时产孢量最高。     

中国弯颈霉培养物的化学成份分析

中国颈霉T.sinense是与冬虫夏草无性型密切相关的真菌。对中国弯颈霉ZN923菌株培养物进行测试,证明其所含化学成分如甘露醇、氨基酸、生物碱、有机酸及甾醇类等与天然冬虫夏草基本相同。红外、紫外吸收图谱也表明两者所含的各类化学成份极为相似。     

辽宁木霉属(Trichoderma)真菌的形态分类研究

对采自辽宁省内14个地方的173份土样和植物组织材料进行分离,获得了54株Trichoderma菌株,采用形态学分类方法鉴定出12种木霉菌,分别是拟康氏木霉(Trichoderma pseudokoningii)、长枝木霉(T.longi-brachiatum)、粘绿木霉(T.virens)、卷曲木霉(T.spirale)、顶孢木霉(T.fertile)、粗壮木霉(T.strigosum)、长孢木霉(T.longipile)、钩状木霉(T.hamatum)、绿色木霉(T.viride)、康氏木霉(T.koningii)、深绿木霉(T.atroviri-de)和哈茨木霉(T.harzianum)。其中长孢木霉为中国新记录种,粗壮木霉和卷曲木霉为东北地区首次报道。文中列有辽宁省木霉属真菌的分种检索表,并附有各木霉菌的生境和分布。     

广西土壤中的暗色丝孢菌Ⅰ(英文)

从广西壮族自治区不同生态环境中采集土壤样品53份,采用土壤平板法和稀释法,从中分离出78个暗色丝孢菌分离物,经鉴定分属20属,33种。其中,弯孢耳孢霉Codinaea curvispora、广西腐质霉Humicola guangxiensis为新种。模式及其他研究过的标本(干制培养物)及活菌种保存在山东农业大学植物病理学标本室(HSAUP),等模式标本(干制培养物)存放在中国科学院真菌标本馆(HMAS)。     

Controlled mycorrhizal initiation as a means to improve root development in somatic embryo plantlets of hybrid larch (Larix × eurolepis)

This study represents the first report of in vitro mycorrhization of somatic embryo-derived plantlets. Effects of fungi on the rooting of plantlets as well as on their ultra-structure were studied. Embryo-derived plantlets of a hybrid larch ( Larix × eurolepis Henry) were grown aseptically in the presence of four ectomycorrhizal fungi. One month after inoculation with Laccaria laccata, Hebeloma cylindrosporum and Pisolithus tinctorius , the number of plantlets which had developed a well-growing root system was significantly improved compared with controls. Furthermore, root branching was strongly stimulated by L. laccata and H. cylindrosporum , which were more efficient species than P. tinctorius and Suillus grevillei . Development of the root system was concomitantly accompanied by an enhancement of shoot growth, except for plantlets inoculated with S. grevillei , which died after three to six months. At the ultrastructural level, cross-sections of lateral roots made six months after inoculation, showed the presence of a Hartig net in the case of L. laccata and H. cylindrosporum , whereas no such net was observed with the two other fungi. With S. grevillei , some perforating hyphae penetrated the cortical cells and resulted in cell death. Evidence of a relationship between structure and efficiency of the association was found.     

Mortality of Delia floralis, Galleria mellonella and Mamestra brassicae treated with insect pathogenic hyphomycetous fungi

Abstract:  The susceptibility of Delia floralis eggs, neonates and larvae and the susceptibility of Galleria mellonella and Mamestra brassicae larvae to seven different Norwegian isolates of the insect pathogenic, hyphomycetous fungi Tolypocladium cylindrosporum , Metarhizium anisopliae and Beauveria bassiana , were investigated. Metarhizium anisopliae isolate ARSEF 5520 was highly virulent to G. mellonella larvae and caused 100% mortality when tested at a concentration of 3.6 × 10⁶ conidia/ml. The same M. anisopliae isolate was not virulent to D. floralis larvae. Isolates of T.cylindrosporum , were equally virulent to G. mellonella and D. floralis causing up to 36.0% mortality of larvae. It is suspected, however, that the use of grated rutabaga as a food source in the D. floralis bioassay reduced the fungal virulence of both M. anisopliae and T. cylindrosporum to D. floralis . Among three T. cylindrosporum isolates tested at a concentration of 1.0 × 10⁷ conidia/ml against eggs of D. floralis none of them reduced the hatching percentage. One isolate, ARSEF 5525 did, however, significantly reduce the longevity of neonates. Beauveria bassiana isolates ARSEF 5510 and ARSEF 5370 tested at a concentration of 1.0 × 10⁷ conidia/ml resulted in M. brassicae mortality levels of 70.0 and 55.0%, respectively. The B. bassiana isolate ARSEF 5557, however, was not virulent to M. brassicae . Among the three isolates tested against M. brassicae the two virulent isolates produced a red pigment, probably oosporein, when cultured in Sabouraud dextrose agar.     

Induced defence responses limit Hartig net formation in ectomycorrhizal birch roots

Roots of clonal birches ( Betula pendula ) were inoculated with the ectomycorrhizal fungi Paxillus involutus (isolates P0 and Mi) and Hebeloma cylindrosporum (strains D1 and D105). These fungi showed different rates of mycorrhiza formation in vitro . Mature mycorrhizas were obtained after only 2–4 d with H. cylindrosporum , whereas 6–8 d were necessary with P. involutus isolate P0, and P. involutus isolate Mi was not able to form mature mycorrhiza during the 10 d of the experiment. Temporal changes in PAL activity and the expression of genes encoding intracellular pathogenesis-related proteins were followed after inoculating birch roots with these fungi. Transient increase of PAL activity, and transient induction of expression of the wound-inducible Bet v1-SC1 gene, were observed in roots challenged with both H. cylindrosporum strains and the P. involutus isolate P0. These changes were found to coincide with hyphal penetration between root cells during Hartig net formation, and were never observed in roots inoculated with the poorly aggressive P. involutus isolate Mi. Examination of mycorrhizal root sections under u.v. light indicated the presence of phenolic compounds in the host cell walls at the vicinity of the Hartig net. These results strongly suggest that hyphal penetration between the root cells triggers a transient defence response which, in turn, could limit Hartig net formation to the outer layer of the root cortex.     

rRNA sequence comparison of Beauveria bassiana, Tolypocladium cylindrosporum, and Tolypocladium extinguens

Five strains of Tolypocladium cylindrosporum, one strain of Tolypocladium extinguens, and nine strains of Beauveria bassiana were analyzed using a rapid rRNA sequencing technique. The sequences of two highly variable domains (D1 and D2) located at the 5' end of the 28S-like rRNA molecule were determined. The phylogenetic tree computed from the absolute number of nucleotide differences shows the separation between the genus Beauveria and the genus Tolypocladium and points out that T. cylindrosporum and T. extinguens probably do not belong to the same genus.     

Peptide uptake in the ectomycorrhizal fungus Hebeloma cylindrosporum: characterization of two di- and tripeptide transporters (HcPTR2A and B)

Constraints on plant growth imposed by low availability of nitrogen are a characteristic feature of ecosystems dominated by ectomycorrhizal plants. Ectomycorrhizal fungi play a key role in the N nutrition of plants, allowing their host plants to access decomposition products of dead plant and animal materials. Ectomycorrhizal plants are thus able to compensate for the low availability of inorganic N in forest ecosystems. The capacity to take up peptides, as well as the transport mechanisms involved, were analysed in the ectomycorrhizal fungus Hebeloma cylindrosporum. The present study demonstrated that H. cylindrosporum mycelium was able to take up di- and tripeptides and use them as sole N source. Two peptide transporters (HcPTR2A and B) were isolated by yeast functional complementation using an H. cylindrosporum cDNA library, and were shown to mediate dipeptide uptake. Uptake capacities and expression regulation of both genes were analysed, indicating that HcPTR2A was involved in the high-efficiency peptide uptake under conditions of limited N availability, whereas HcPTR2B was expressed constitutively.     

Characterization of the ER-located zinc transporter ZnT1 and identification of a vesicular zinc storage compartment in Hebeloma cylindrosporum

Metal tolerance of filamentous fungi is a poorly understood mechanism. In order to unravel the molecular basis of zinc (Zn) tolerance in the ectomycorrhizal fungal model Hebeloma cylindrosporum, we carried out a functional screening of an H. cylindrosporum cDNA library in the zrc1Δ mutant strain of Saccharomyces cerevisiae to search for genes conferring Zn tolerance to yeast cells. This strategy allowed the isolation of HcZnT1, a gene belonging to the cation diffusion facilitator family, which induced tolerance to Zn, but not to other metals. HcZnT1 was constitutively expressed in Hebeloma cells, whatever the Zn status of the medium and the fungal cell type (mycelia, sporocarps, mycorrhizas). A HcZnT1:GFP fusion protein was expressed in yeast and the corresponding fluorescence was recorded on endoplasmic reticulum membranes. Taken together, these different findings suggest a dual role of HcZnT1 in Zn homeostasis of fungal cells, by supplying requested Zn ions for the functioning of the endoplasmic reticulum as well as by detoxifying the cytosol under Zn stress. Zn pools were also investigated by using the Zn-specific fluorophore zinquin in H. cylindrosporum cells. Zinquin labeling revealed compartmentalization in intracellular vesicles interspersed throughout the cytoplasm that do not correspond to vacuolar compartments. Altogether the present data represent the first steps into the understanding of Zn homeostasis and tolerance in Hebeloma.     

The auxin-inducible GH3 homologue Pp-GH3.16 is downregulated in Pinus pinaster root systems on ectomycorrhizal symbiosis establishment

In an attempt to determine whether auxin-regulated plant genes play a role in ectomycorrhizal symbiosis establishment, we screened a Pinus pinaster root cDNA library for auxin-upregulated genes. This allowed the identification of a cDNA, Pp-GH3.16, which encodes a polypeptide sharing extensive homologies with GH3 proteins of different plants. Pp-GH3.16 was specifically upregulated by auxins and was not affected by cytokinin, gibberellin, abscisic acid or ethylene, or by heat shock, water stress or anoxia. Pp-GH3.16 mRNAs were quantified in pine roots inoculated with two ectomycorrhizal fungi, Hebeloma cylindrosporum and Rhizopogon roseolus. Surprisingly, Pp-GH3.16 was downregulated following inoculation with both fungal species. The downregulation was most rapid on establishment of symbiosis with an indole-3-acetic acid (IAA)-overproducing mutant of H. cylindrosporum, which overproduced mycorrhizas characterized by a hypertrophic Hartig net. This indicates that, despite being auxin-inducible, Pp-GH3.16 can be downregulated on establishment of symbiosis with a fungus that releases auxin. By contrast, Pp-GH3.16 was not downregulated in pine root systems inoculated with a nonmycorrhizal mutant of H. cylindrosporum, suggesting that the downregulation we observed in mycorrhizal root systems was a component of the molecular cross-talk between symbiotic partners at the origin of differentiation of symbiotic structures.