The potential saprotrophic capacity of foliar endophytic fungi from Quercus gambelii

Endophytic fungi occur in living tissues of terrestrial plants. Many of these fungi are primarily biotrophic, but the trophic range of endophytic fungi as a group may not be fully appreciated. In this study, our goals were (1) for the Class 3 foliar endophytic fungi isolated from Quercus gambelii, determine their potential saprotrophic capacity, which we define as the difference in growth rate in culture on Quercus gambelii leaf litter medium and control medium lacking leaf litter and (2) quantify sources of variation among isolates of these endophytic fungi in potential saprotrophic capacity, including variation due to microsite within host trees (leaves receiving full sun vs. shade) and variation within and among fungal genera. We found that 48 of the 49 tested endophytic fungal isolates have significant potential saprotrophic capacity. Contrary to expectation, the amount of solar radiation available to the leaf from which the fungi were isolated had no significant impact on potential saprotrophic capacity and there was more variability in potential saprotrophic capacity among isolates within a genus than among genera. Our results suggest that some Class 3 endophytic fungi may have the potential to function as saprotrophic fungi within plant litter, but this remains to be seen for these Quercus gambelii isolates under more natural circumstances.     

Community Analysis Reveals Close Affinities Between Endophytic and Endolichenic Fungi in Mosses and Lichens

Endolichenic fungi live in close association with algal photobionts inside asymptomatic lichen thalli and resemble fungal endophytes of plants in terms of taxonomy, diversity, transmission mode, and evolutionary history. This similarity has led to uncertainty regarding the distinctiveness of endolichenic fungi compared with endophytes. Here, we evaluate whether these fungi represent distinct ecological guilds or a single guild of flexible symbiotrophs capable of colonizing plants or lichens indiscriminately. Culturable fungi were sampled exhaustively from replicate sets of phylogenetically diverse plants and lichens in three microsites in a montane forest in southeastern Arizona (USA). Intensive sampling combined with a small spatial scale permitted us to decouple spatial heterogeneity from host association and to sample communities from living leaves, dead leaves, and lichen thalli to statistical completion. Characterization using data from the nuclear ribosomal internal transcribed spacer and partial large subunit (ITS-LSU rDNA) provided a first estimation of host and substrate use for 960 isolates representing five classes and approximately 16 orders, 32 families, and 65 genera of Pezizomycotina. We found that fungal communities differ at a broad taxonomic level as a function of the phylogenetic placement of their plant or lichen hosts. Endolichenic fungal assemblages differed as a function of lichen taxonomy, rather than substrate, growth form, or photobiont. In plants, fungal communities were structured more by plant lineage than by the living vs. senescent status of the leaf. We found no evidence that endolichenic fungi are saprotrophic fungi that have been “entrapped” by lichen thalli. Instead, our study reveals the distinctiveness of endolichenic communities relative to those in living and dead plant tissues, with one notable exception: we identify, for the first time, an ecologically flexible group of symbionts that occurs both as endolichenic fungi and as endophytes of mosses.     

Colonization and lignin decomposition of Camellia japonica leaf litter by endophytic fungi

Endophytic fungi occur on various types of leaf litter, but few studies have been done on their roles as saprophytes in decomposition. This study examined the succession of fungi in live, newly shed, and decomposing leaves at 2 months of decomposition of Camellia japonica and chemical changes in decomposing leaves colonized by endophytes. Coccomyces nipponicum, Lophodermium sp., Geniculosporium sp. 1, and Colletotrichum gloeosporioides were isolated from living leaves. Coccomyces nipponicum and Lophodermium sp. were also isolated frequently from newly shed and decomposing leaves. These two fungi caused a decrease of lignin content and bleaching in decomposing leaves under field and laboratory conditions. Total hyphal length in decomposing leaves was higher in bleached portions than in surrounding nonbleached portions, which probably reflected the early onset of hyphal growth of endophytes inside leaf tissue at leaf senescence or death. Incubation of newly shed leaves that were sterilized to exclude previously established endophytes resulted in no occurrence of bleached portions in decomposing leaves on the forest floor. This result indicated that these endophytes were incapable of colonizing leaves directly after litterfall and that the persistence of endophytes from live leaves was crucial for their colonization in decomposing leaves.     

Fungal diversity in galls of baldcypress trees

The baldcypress midge (Taxodiomyia cupressi and Taxodiomyia cupressiananassa) forms a gall that originates from leaf tissue. Female insects may inoculate galls with fungi during oviposition, or endophytes from the leaf tissue may grow into the gall interior. We investigated fungal diversity inside of baldcypress galls, comparing the gall communities to leaves and comparing fungal communities in galls that had successful emergence versus no emergence of midges or parasitoids. Galls of midges that successfully emerged were associated with diverse gall fungal communities, some of which were the same as the fungi found in surrounding leaves. Galls with no insect emergence were characterized by relatively low fungal diversity.     

A Phylogenetic Evaluation of Whether Endophytes Become Saprotrophs at Host Senescence

Fungal endophytes and saprotrophs generally play an important ecological role within plant tissues and dead plant material. Several reports based solely on morphological observations have postulated that there is an intimate link between endophytes and saprotrophs. This study aims to provide valuable insight as to whether some endophytic fungi manifest themselves as saprotrophs upon host decay. Ribosomal DNA-based sequence comparison and phylogenetic relationships from 99 fungal isolates (endophytes, mycelia sterilia, and saprotrophs) recovered from leaves and twigs of Magnolia liliifera were investigated in this study. Molecular data suggest there are fungal taxa that possibly exist as endophytes and saprotrophs. Isolates of Colletotrichum, Fusarium, Guignardia, and Phomopsis, which are common plant endophytes, have high sequence similarity and are phylogenetically related to their saprotrophic counterparts. This provides evidence to suggest that some endophytic species change their ecological strategies and adopt a saprotrophic lifestyle. The implication of these findings on fungal biodiversity and host specificity is also discussed.     

Phyllosphere fungi on living and decomposing leaves of giant dogwood

Phyllosphere fungi on living leaves and their succession on decomposing leaves were studied on giant dogwood (Swida controversa). A total of 12 and 34 fungal species were isolated from the interior and surface, respectively, of living leaves, and 15 frequent species were considered as phyllosphere fungi. Six of these 15 species were also frequent on decomposing litter. Characteristic successional trends were observed in the 6 phyllosphere fungi during decomposition. The sum of frequencies of endophytes decreased as decomposition progressed, and no endophytes were isolated from the litter at the 11th month of decomposition. The sum of frequencies of epiphytes increased as decomposition progressed. Endophytes and epiphytes showed different responses to litter mass loss and concentrations of nitrogen, lignin, and total carbohydrates during the decomposition process. These results suggested that epiphytes may survive on decomposing leaves as primary decomposers on the ground, thereby excluding endophytes by competition for available energy sources, and that epiphytes may have a greater contribution to decomposition than endophytes in dogwood leaves.     

Plant identity strongly structures the root-associated fungal community in a diverse subtropical forest

Revealing the relationship between plants and root-associated fungi is very important in understanding diversity maintenance and community assembly in ecosystems. However, the community assembly of root-associated fungi of focal plant species along a subtropical plant species diversity gradient is less documented. Here, we examined root-associated fungal communities associated with five ectomycorrhizal (EM) plant species (Betula luminifera, Castanea henryi, Castanopsis fargesii, C. sclerophylla, and Quercus serrate) in a Chinese subtropical woody plant species diversity (1, 2, 4, 8, 16 and 24 species) experiment, using paired-end Illumina MiSeq sequencing of the ITS2 region. In total, we detected 1933 root-associated fungal operational taxonomic units (OTUs) at a 97% sequence similarity level. Plant identity had a significant effect on total and saprotrophic fungal OTU richness, but plant species diversity level had a significant effect on saprotrophic and pathogenic fungal OTU richness. The community composition of total, saprotrophic and EM fungi was structured by plant identity and plant species diversity level. However, the community composition of pathogenic fungi was only shaped by plant identity. This study highlights that plant identity has a stronger effect on the root-associated fungal community than plant species diversity level in a diverse subtropical forest ecosystem.     

Fungal endophyte communities in two tropical forests of southern India: diversity and host affiliation

Fifteen tree species from a tropical dry thorn forest and fifteen tree species from a tropical dry deciduous forest in the Mudumalai Wildlife Sanctuary, Nilgiri Biosphere Reserve, southern India, were surveyed for their foliar endophyte communities during the dry and wet seasons. Surface sterilized leaf segments of uniform dimension were plated on nutrient agar and culturable endophytes growing from the segments were identified. Endophyte diversity was greater in the dry thorn forest than in the dry deciduous forest in the dry season. Although the isolation frequency of culturable endophytes increased for both forests during the wet season, the assemblages were represented not by any unique fungal species but by the commonly occurring ones. Furthermore, although individual leaves were densely colonized by endophytes, only a few species of endophytes colonized the whole leaves; and, only a few fungal species dominated the foliar endophytic communities and were common for both forests during both dry and wet seasons. Thus, even under wet conditions that favour dispersal and infection by fungi, the endophyte diversity increased only marginally, an indication that certain tropical forests are not hyperdiverse with reference to fungal endophytes. This should be considered when using culturable endophyte diversity as a surrogate for estimating global fungal diversity.     

Evolutionary histories and mycorrhizal associations of mycoheterotrophic plants dependent on saprotrophic fungi

Mycoheterotrophic plants (MHPs) are leafless, achlorophyllous, and completely dependent on mycorrhizal fungi for their carbon supply. Mycorrhizal symbiosis is a mutualistic association with fungi that is undertaken by the majority of land plants, but mycoheterotrophy represents a breakdown of this mutualism in that plants parasitize fungi. Most MHPs are associated with fungi that are mycorrhizal with autotrophic plants, such as arbuscular mycorrhizal (AM) or ectomycorrhizal (ECM) fungi. Although these MHPs gain carbon via the common mycorrhizal network that links the surrounding autotrophic plants, some mycoheterotrophic lineages are associated with saprotrophic (SAP) fungi, which are free-living and decompose leaf litter and wood materials. Such MHPs are dependent on the forest carbon cycle, which involves the decomposition of wood debris and leaf litter, and have a unique biology and evolutionary history. MHPs associated with SAP fungi (SAP-MHPs) have to date been found only in the Orchidaceae and likely evolved independently at least nine times within that family. Phylogenetically divergent SAP Basidiomycota, mostly Agaricales but also Hymenochaetales, Polyporales, and others, are involved in mycoheterotrophy. The fungal specificity of SAP-MHPs varies from a highly specific association with a single fungal species to a broad range of interactions with multiple fungal orders. Establishment of symbiotic culture systems is indispensable for understanding the mechanisms underlying plant–fungus interactions and the conservation of MHPs. Symbiotic culture systems have been established for many SAP-MHP species as a pure culture of free-living SAP fungi is easier than that of biotrophic AM or ECM fungi. Culturable SAP-MHPs are useful research materials and will contribute to the advancement of plant science.

     

Endophytic fungi increase the processing rate of leaves by leaf‐cutting ants (Atta)

1. Fungal endophytes are microfungi that reside asymptomatically inside of leaf tissues, increasing in density and diversity through time after leaves flush. Previous studies have suggested that the presence of fungal endophytes in the harvest material of leaf‐cutting ants (Atta colombica, Guérin‐Méneville) may negatively affect the ants and their fungal cultivar. 2. In the present study, it was tested whether the presence and diversity of fungal endophytes affected the amount of time necessary for leaf‐cutter ants to cut, process, and plant leaf material in their fungal garden. It was found that ants took 30–43% longer to cut, carry, clean, and plant leaf tissue with high relative to low endophyte abundance, and that the ants responded similarly to leaf tissue with high or low endophyte diversity. 3. It was further investigated whether the fungal cultivars' colonisation rate was greater on leaf material without fungal endophytes. No difference in the ants' cultivar colonisation rate on leaf tissue with high or low endophyte abundance was observed.     

Taxa-area relationship and neutral dynamics influence the diversity of fungal communities on senesced tree leaves

This study utilized individual senesced sugar maple and beech leaves as natural sampling units within which to quantify saprotrophic fungal diversity. Quantifying communities in individual leaves allowed us to determine if fungi display a classic taxa–area relationship (species richness increasing with area). We found a significant taxa–area relationship for sugar maple leaves, but not beech leaves, consistent with Wright's species‐energy theory. This suggests that energy availability as affected plant biochemistry is a key factor regulating the scaling relationships of fungal diversity. We also compared taxa rank abundance distributions to models associated with niche or neutral theories of community assembly, and tested the influence of leaf type as an environmental niche factor controlling fungal community composition. Among rank abundance distribution models, the zero‐sum model derived from neutral theory showed the best fit to our data. Leaf type explained only 5% of the variability in community composition. Habitat (vernal pool, upland or riparian forest floor) and site of collection explained > 40%, but could be attributed to either niche or neutral processes. Hence, although niche dynamics may regulate fungal communities at the habitat scale, our evidence points towards neutral assembly of saprotrophic fungi on individual leaves, with energy availability constraining the taxa–area relationship.     

Sapwood and heartwood affect differentially bacterial and fungal community structure and successional dynamics during Quercus petraea decomposition

In forests, bacteria and fungi are key players in wood degradation. Still, studies focusing on bacterial and fungal successions during the decomposition process depending on the wood types (i.e. sapwood and heartwood) remain scarce. This study aimed to understand the effect of wood type on the dynamics of microbial ecological guilds in wood decomposition. Using Illumina metabarcoding, bacterial and fungal communities were monitored every 3 months for 3 years from Quercus petraea wood discs placed on forest soil. Wood density and microbial enzymes involved in biopolymer degradation were measured. We observed rapid changes in the bacterial and fungal communities and microbial ecological guilds associated with wood decomposition throughout the experiment. Bacterial and fungal succession dynamics were very contrasted between sapwood and heartwood. The initial microbial communities were quickly replaced by new bacterial and fungal assemblages in the sapwood. Conversely, some initial functional guilds (i.e. endophytes and yeasts) persisted all along the experiment in heartwood and finally became dominant, possibly limiting the development of saprotrophic fungi. Our data also suggested a significant role of bacteria in nitrogen cycle during wood decomposition.     

Diversity and decomposition potential of endophytes in leaves of a Cinnamomum camphora plantation in China

This study was carried out to improve our understanding of the diversity and decomposition potential of endophytes in the leaves of Cinnamomum camphora trees grown in a subtropical region of China. We isolated and identified endophytic fungi from senescent leaves of C. camphora and tested their role in decomposition through pure-culture and pre-colonization. A total of 2,861 endophytic fungi isolated from 69 leaves of C. camphora were grouped into 39 taxa comprising 36 Ascomycetes and 3 Basidiomycetes based on sporulation and ITS sequence analysis. Of these, Colletotrichum gloeosporioides was the most common species (69% relative abundance and 96% colonization frequency), followed by Cladosporium sp.1, Colletotrichum sp. and Chaetomium sp. All 39 endophytes had the ability to decompose C. camphora leaf litter in pure culture, and a few exhibited >20% litter mass loss in 2 months. In most cases, single endopyhytic species showed lower mass loss than mixed microbial groups from active soil after 60 or 120 days. In pre-inoculation, endophytic fungi like Chaetomium sp., Cladosporium sp.1, C. gloeosporioides, Colletotrichum sp. and Guignardia sp. exhibited higher abundance and caused greater mass loss, indicating the potential of these groups to enter and significantly accelerate the process of decomposition. This study concludes that, after entering the decomposition process, selected endophytic fungi with high abundance could influence significantly the decomposition process and thus probably affect carbon and nutrient cycling in C. camphora plantations.     

Fungal endophytes and phytochemistry of oak foliage: determinants of oviposition preference of leafminers?

Sedentary insect herbivores, such as gallformers and leafminers, are usually non-randomly distributed among and within host plants. Dispersion of these insects is largely a function of female oviposition choice. In field experiments and observations spanning two growing seasons, we tested the hypothesis that selective oviposition on individual leaves within trees by the dominant herbivore of Emory oak, the monophagous leaf-miner Cameraria sp. nov., is determined by the probability of colonization by endophytic fungi. These fungi are alleged to act as plant mutualists by deterring, killing, or inhibiting the growth of insect herbivores. We found that leaves selected by females for oviposition and paired, unmined leaves were equally likely to be colonized by fungal endophytes. Furthermore, condensed and hydrolyzable tannin levels, purported inhibitors of fungal infection, and protein content did not vary between leaves selected by females and unmined leaves, or between leaves with and without endophyte infections. We conclude that female Cameraria do not choose leaves within trees for oviposition on the basis of propensity for endophytic fungal infection or on phytochemical parameters that might indicate probability of future infections. At this spatial scale at least, fungal endophytes do not explain the highly aggregated distribution of Cameraria among leaves and associated costs in terms of increased larval mortality. Fungal endophytes may, nevertheless, affect leafminer dispersion and abundance at larger spatial scales, such as host plant populations or species. We did find, however, that the amount of mining activity on leaves is positively associated with increased colonization by fungal endophytes. We suggest that mining activity increases endophyte fungal infections by facilitating spore germination and hyphal penetration into the leaf or by altering leaf phytochemistry. The facilitation of endophyte colonization by leafmining activity coupled with the lack of predictability of endophyte infections based on leaf phytochemistry and almost 100% infectivity of all oak leaves during sporadic wet years may prevent female leafminers from discriminating leaves for oviposition on the basis of current or future levels of endophytes in leaves.     

Mutualistic interactions on a knife-edge between saprotrophy and pathogenesis

Saprophytic, ectomycorrhizal (ECM) and pathogenic fungi play a key role in carbon and nutrient cycling in forest ecosystems. Whereas more than 50 genomes of saprotrophic and pathogenic fungi have been published, only two genomes of ECM fungi, Laccaria bicolor and Tuber melanosporum, have been released. Comparative analysis of the genomes of biotrophic species highlighted convergent evolution. Mutualistic and pathogenic biotrophic fungi share expansion of genome size through transposon proliferation and common strategies to avoid plant detection. Differences mainly rely on nutritional strategies. Such analyses also pinpointed how blurred the molecular boundaries are between saprotrophism, symbiosis and pathogenesis. Sequencing of additional ECM species, as well as soil saprotrophic fungi, will facilitate the identification of conserved traits for ECM symbiosis and those leading to the transition from white-rotting and brown-rotting to the ECM lifestyle.     

Fungal Epiphytes and Endophytes of Coffee Leaves (Coffea arabica)

Plants harbor diverse communities of fungi and other microorganisms. Fungi are known to occur both on plant surfaces (epiphytes) and inside plant tissues (endophytes), but the two communities have rarely been compared. We compared epiphytic and endophytic fungal communities associated with leaves of coffee (Coffea arabica) in Puerto Rico. We asked whether the dominant fungi are the same in both communities, whether endophyte and epiphyte communities are equally diverse, and whether epiphytes and endophytes exhibit similar patterns of spatial heterogeneity among sites. Leaves of naturalized coffee plants were collected from six sites in Puerto Rico. Epiphytic and endophytic fungi were isolated by placing leaf pieces on potato dextrose agar without and with surface sterilization, respectively. A total of 821 colonies were isolated and grouped into 131 morphospecies. The taxonomic affinities of the four most common nonsporulating fungi were determined by sequencing the nuclear ribosomal internal transcribed spacer (ITS) region: two grouped with Xylaria and one each with Botryosphaeria and Guignardia. Of the most common genera, Pestalotia and Botryosphaeria were significantly more common as epiphytes; Colletotrichum, Xylaria, and Guignardia were significantly more common as endophytes. Suprisingly, more morphospecies occurred as endophytes than as epiphytes. Differences among sites in number of fungi per plant were significant. Thus epiphytic and endophytic communities differed greatly on a single leaf, despite living only millimeters apart, and both communities differed from site to site. Significant correlations between occurrence of fungal morphospecies suggested that fungi may have positive or negative effects on their neighbors. This is the first quantitative comparison of epiphytic and endophytic fungal floras in any plant, and the first to examine endophytic fungi or epiphytic fungi in leaves of coffee, one of the world’s most valuable crops.     

Relationships between the litter colonization by saprotrophic and arbuscular mycorrhizal fungi with depth in a tropical forest

Fungal colonization of litter has been described mostly in terms of fructification succession in the decomposition process or the process of fungal ligninolysis. No studies have been conducted on litter colonization by arbuscular mycorrhizal fungi (AMF) and their relationship with the presence of saprotrophic fungi. The aim of the present study was to evaluate the relationships that exist in simultaneous leaf litter colonization by AMF and saprotrophic fungi and the relationships between rates of litter and associated root colonization by AMF at different soil depths. We selected Eugenia sp. and Syzygium sp. in a riparian tropical forest, with an abundant production of litter (O horizon), we evaluated litter and root colonization at different depths, its C:N ratios, and the edaphic physico-chemical parameters of the A horizon immediately below the litter layer. Litter colonization by saprotrophic fungi and AMF increased with depth, but the saprotrophic fungal colonization of some litter fragments decreased in the lowermost level of the litter while AMF litter colonization continued to increase. Plant roots were present only in the middle and bottom layers, but their mycorrhizal colonization did not correlate with litter colonization. The external hyphae length of AMF is abundant (ca. 20 m g(-1) sample) and, in common with sample humidity, remained constant with increasing depth. We conclude that in zones of riparian tropical forest with abundant sufficient litter accumulation and abundant AMF external hyphae, the increase in litter colonization by AMF with depth correlates to the colonization by saprotrophic fungi, but their presence in the deepest layers is independent of both litter colonization by saprotrophic fungi and root colonization by AMF.     

广藿香内生真菌多样性及其对青枯菌的拮抗活性

【目的】对广藿香各部位内生真菌的类群结构及多样性进行系统分析,筛选出对青枯菌具拮抗活性的菌株。【方法】采用组织块分离法从广藿香健康植株的根、茎、叶中分离内生真菌,结合形态学和基于ITS-r DNA序列分析鉴定方法分析广藿香内生真菌的类群结构及多样性,并利用双层平板拮抗法筛选对青枯菌具有抑制活性的菌株。【结果】从广藿香中共分离获得313株内生真菌,隶属于30个属,其中链格孢属(占28.75%)、拟茎点霉属(占23.00%)和炭疽菌属(11.82%)为优势类群,此外还分离到炭皮菌属、弯孢聚壳属、Gibellulopsis、新萨托菌属、葡萄座腔菌属、篮状菌属等较为少见的类群。茎、叶内生真菌的定殖率和分离率明显高于根,而根部多样性指数为2.64,要高于茎(2.00)和叶(1.97);广藿香茎与叶、根与茎、根与叶之间的内生真菌相似性指数分别为0.35、0.20、0.19,均小于0.5,显示各部位之间的内生真菌组成不相似程度高。从313株广藿香内生真菌中通过拮抗实验筛选到16株对青枯菌有拮抗活性的菌株,其中GHXR07、GHXR27和GHXR29的拮抗活性尤为显著,分别被鉴定为Talaromyces sp.、Myrothecium roridum Tode和Talaromyces wortmannii(Kl?cker)Benjamin。【结论】广藿香内生真菌具有丰富的物种多样性且其类群分布具有一定的组织特异性,其中部分菌株对青枯菌具有明显的拮抗活性。     

Diversity and host recurrence of fungi associated with the bleached leaf litter in a subtropical forest

Fungi that selectively remove lignin cause extensive bleaching of leaf litter, which results in the acceleration of litter decomposition. The taxonomic diversity and host recurrence of saprotrophic fungi associated with bleached leaf litter were investigated in a subtropical forest in southern Japan. A total of 211 fungal isolates and sporocarps were obtained from bleached portions of leaf litter of 33 plant species in 18 families and were classified into 83 operational taxonomic units (OTUs) at the 97% similarity level of the ITS rDNA sequence. These fungal OTUs belonged to Rhytismataceae and Xylariaceae in the Ascomycota, and Marasmiaceae, Mycenaceae, Lachnocladiaceae, and Physalacriaceae in the Basidiomycota. OTUs in Rhytismataceae and Marasmiaceae showed a significantly higher degree of recurrence for plant species than simulated networks where partners were associated randomly. In contrast, OTUs in Xylariaceae and Mycenaceae showed no significant recurrence for plant species. Such differing degrees of recurrence for plant species implied different roles of fungal families in leaf litter decomposition.     

Effects of Riparian Plant Diversity Loss on Aquatic Microbial Decomposers Become More Pronounced with Increasing Time

We examined the potential long-term impacts of riparian plant diversity loss on diversity and activity of aquatic microbial decomposers. Microbial assemblages were obtained in a mixed-forest stream by immersion of mesh bags containing three leaf species (alder, oak and eucalyptus), commonly found in riparian corridors of Iberian streams. Simulation of species loss was done in microcosms by including a set of all leaf species, retrieved from the stream, and non-colonized leaves of three, two or one leaf species. Leaves were renewed every month throughout six months, and microbial inoculum was ensured by a set of colonized leaves from the previous month. Microbial diversity, leaf mass loss and fungal biomass were assessed at the second and sixth months after plant species loss. Molecular diversity of fungi and bacteria, as the total number of operational taxonomic units per leaf diversity treatment, decreased with leaf diversity loss. Fungal biomass tended to decrease linearly with leaf species loss on oak and eucalyptus, suggesting more pronounced effects of leaf diversity on lower quality leaves. Decomposition of alder and eucalyptus leaves was affected by leaf species identity, mainly after longer times following diversity loss. Leaf decomposition of alder decreased when mixed with eucalyptus, while decomposition of eucalyptus decreased in mixtures with oak. Results suggest that the effects of leaf diversity on microbial decomposers depended on leaf species number and also on which species were lost from the system, especially after longer times. This may have implications for the management of riparian forests to maintain stream ecosystem functioning.