福廷瓶头霉与马尾松的共生特征

深色有隔内生真菌（dark septate endophyte,DSE）广泛定殖于植物根系,对促进植物生长、提高植物抗逆能力具重要作用。本研究从马尾松Pinus massoniana根系分离到一株DSE,于无菌条件下研究了此菌与马尾松的共生特征,研究结果表明：基于形态学和分子生物学分析,此菌被鉴定为福廷瓶头霉Phialocephala fortinii,在PDA培养基25℃培养条件下,此菌不产孢,菌丝深色、具隔,最适培养基为PDA。此菌与马尾松根系共生体的形成过程可划分为3个时期,即侵入前期：接种后2d,新生菌丝向马尾松根系定向生长并开始接触根系,但未侵入根系内部;侵入期：接种后4d,菌丝与马尾松根系接触侵入根内并在根系皮层细胞间扩展延伸;形成期：接种后6d,菌丝继续在马尾松根系内扩展并形成富含脂类物质的微菌核。接种福廷瓶头霉显著增加了马尾松生物量的积累（P<0.05）,并影响了其根系发育。接种后,马尾松主根生长受限,侧根生长显著提高（P<0.05）,侧根长度较未接种处理增加了112.87%。被侵染根系的根毛数量减少,并由大量根外菌丝包裹。以上结果对进一步揭示DSE与宿主的共生机理有一定指导意义。     

New cytotoxic metabolites from a deep-sea-derived fungus, Phialocephala sp., strain FL30r

Two new sorbicillinoids, 1 and 2 , together with a novel benzofuranone derivative named phialofurone ( 3 ), were isolated from a deep‐sea sediment‐derived fungus, Phialocephala sp. Their structures were established on the basis of spectroscopic data. All compounds displayed cytotoxic effects against P388 (IC₅₀ values of 11.5±1.4, 0.1±0.1, and 0.2±0.01 μM , resp.) and K562 (IC₅₀ values of 22.9±0.8, 4.8±0.3 and 22.4±0.9 μM , resp.) cell lines.     

Effects of isolates of ectomycorrhizal fungi and endophytic Mycelium radicis atrovirens that were dominant in soil from disturbed sites on growth of Betula platyphylla var. japonica seedlings

Effects of ectomycorrhizal fungi and endophytic Mycelium radicis atrovirens Melin (MRA) on growth of Betula platyphylla var. japonica seedlings were investigated under aseptic culture conditions. Three isolates of ectomycorrhizal fungi and two isolates of MRA were used. One MRA isolate was Phialocephala fortinii. Previous field work revealed that these isolates were dominant on the roots of B. platyphylla var. japonica seedlings grown in a mineral subsoil that had been exposed by the removal of surface soil. After a 100-day incubation, the growth of the seedlings was significantly enhanced by the colonization of these ectomycorrhizal fungal isolates as compared with uninoculated seedlings. In contrast, the growth of seedlings was retarded by the colonization of the MRA isolates. The growth of seedlings that were co-inoculated with ectomycorrhizal fungi and MRA was similar to that of uninoculated seedlings in most cases. These results suggest that ectomycorrhizal fungi have a beneficial effect on the growth of B. platyphylla var. japonica seedlings and that they suppress the deleterious effect of MRA. Thus, these ectomycorrhizal fungi probably have an important role in establishing B. platyphylla var. japonica seedlings during the initial stage of re-vegetation following site disturbance by the removal of surface soil.     

Monitoring the spatial and temporal dynamics of a community of the tree-root endophyte Phialocephala fortinii s.l

Phialocephala fortinii sensu lato was isolated from Picea abies roots that had been collected from the same 3 x 3-m forest plot in 2001 and 2004, to examine the spatial and temporal dynamics of this fungal community. RFLP analysis was used to define the multilocus haplotype (MLH) of each isolate. Pielou's measure of association and chi(2) tests of independence were employed to examine the randomness of patterns of spatial distribution of MLH observed in 2001 and 2004. Population differentiation between the two samplings was tested using the G(ST) statistic. In 2001, 144 strains of 28 MLH were isolated; in 2004, 139 strains of 29 MLH. Abundant MLH in 2001 also prevailed in 2004, and the same two cryptic species were dominant in both collections. The probability of being isolated in both years increased with increasing frequency of an MLH. The patterns of spatial distribution of most MLH did not differ between years. The G(ST) values indicated identity of the two collections. Communities of P. fortinii sensu lato remain spatially and genetically stable for at least 3 yr.     

Phialocephala fortinii increases aluminum tolerance in Miscanthus sinensis growing in acidic mine soil

Miscanthus sinensis growing in our study mine site contained a high concentration of Al in the adventitious roots. It has a root endophyte, Phialocephala fortinii, in its adventitious roots at a high frequency. The purpose of this study was to elucidate the effects of P. fortinii on Al tolerance mechanisms of M. sinensis and reveal potential underlying mechanisms. In the absence of P. fortinii, M. sinensis produced chlorogenic, citric, and malic acids that could act to alleviate Al toxicity in acidic mine soil. Up on fungal inoculation, the levels of these compounds were reduced, although the growth of seedlings and Mg concentration in the roots were increased. IAA production by the fungus may contribute to enhanced plant growth whereas an increase of Mg uptake could reduce toxicity of reactive oxygen species under Al stress. These actions of P. fortinii could promote growth and survival of M. sinensis in mine sites.     

Microsatellite size homoplasies and null alleles do not affect species diagnosis and population genetic analysis in a fungal species complex

The suitability of 13 microsatellite loci for species diagnosis and population genetics in 11 species of the Phialocephala fortinii s.l.-Acephala applanata species complex (PAC) was assessed. Two data sets were compared to test possible biases in species typing and clone detection resulting from null alleles and size homoplasies. The first data set was based on fragment lengths derived from a multiplex polymerase chain reaction (PCR) assay and the second data set was received from singleplex PCR at lower stringency and sequencing. Most null alleles observed in the multiplex PCR assay could be amplified during singleplex PCR under less stringent conditions. Size homoplasies resulting from mutations in flanking regions and differences in microsatellite structures were observed. For example, Phialocephala uotolensis possessed a (CT)(13) in addition to the (GT)(x) motif at locus mPF_0644. Despite the occurrence of null alleles and size homoplasies, species diagnosis and population genetic analysis studies were not affected. These markers will facilitate studies on population biology, ecology and biogeography of PAC species.     

Spatial distribution of discrete RAPD phenotypes of a root endophytic fungus, Phialocephala fortinii, at a primary successional site on a glacier forefront

Phialocephala fortinii is among the few identified hyphomycetes belonging to the Mycelium radicis atrovirens complex. This 'dark-septate endophyte' has a global distribution and colonizes a wide variety of host plants. In this study, the spatial distribution of discrete genets of P. fortinii on the forefront of a receding glacier was analysed using the randomly amplified polymorphic DNA (RAPD) technique to determine plants colonized and patterns of colonization. In two consecutive years of sampling, a total of 74 isolates of P. fortinii were obtained from nine plant species, typically ectomycorrhizal, ericoid mycorrhizal or non-mycorrhizal. The isolates showed substantial variation, sharing on average approx. half the RAPD markers. In the first year, three isolates belonging to a single genet were obtained from two plants separated by a distance of nearly 1.5 m. The continuity of this genet was assessed by a sampling the following year. No isolates similar to that, or any of the genets collected the year before were observed. Consequently, the identical isolates from the previous year were concluded to represent discontinuous ramets. Two additional large genets were observed during the second year of sampling, which inhabited roots of several plants representing three different species. These data suggest that the sharing of P. fortinii genets among plant species might play a fundamental role in adaptation and interaction within the whole plant community in a system undergoing primary succession.