Temporal variation in temperature and rainfall differentially affects ectomycorrhizal colonization at two contrasting sites

We examined the roles that seasonal shifts in precipitation and temperature played in the ectomycorrhizal (ECM) colonization of pinyon pine ( Pinus edulis Engelm.) at two contrasting sites in northern Arizona. Pinyons growing in ash and cinder soils experienced much greater water and nutrient stress than pinyons growing nearby in sandy-loam soils. Over a one year period, we obtained monthly measurements of ECM colonization, root zone soil moisture and temperature, and air temperature and precipitation. Four major patterns emerged. Firstly, although climate as measured by ambient temperature and precipitation did not vary between the two sites, soil temperature was significantly higher and soil moisture significantly lower at the cinder site than at the sandy-loam site. Secondly, ECM colonization was significantly higher at the cinder site for 5 of 12 months. Thirdly, although nearly 70% of the variation in ECM colonization of pinyons growing in cinder soil was predicted by a combination of soil moisture and soil temperature, these same variables had little predictive power for pinyons growing in sandy-loam soils. Air temperature and precipitation were also significantly correlated with ECM colonization at the cinder site but not the sandy-loam site. Fourthly, a watering experiment showed that ECM colonization significantly increased with supplemental water at the cinder site, but not at the sandy-loam site. Thus, in two sites that did not differ in plant community or climate, ectomycorrhizas in cinder soils were far more sensitive to changes in moisture and temperature than ectomycorrhizas in sandy-loam soils.     

Ostryopsis davidiana seedlings inoculated with ectomycorrhizal fungi facilitate formation of mycorrhizae on Pinus tabulaeformis seedlings

Reforestation in China is important for reversing anthropogenic activities that degrade the environment. Pinus tabulaeformis is desired for these activities, but survival and growth of seedlings can be hampered by lack of ectomycorrhizae. When outplanted in association with Ostryopsis davidiana plants on reforestation sites, P. tabulaeformis seedlings become mycorrhizal and survival and growth are enhanced; without O. davidiana, pines often remain without mycorrhizae and performance is poorer. To better understand this relationship, we initiated an experiment using rhizoboxes that restricted root and tested the hypothesis that O. davidiana seedlings facilitated ectomycorrhizae formation on P. tabulaeformis seedlings through hyphal contact. We found that without O. davidiana seedlings, inocula of five indigenous ectomycorrhizal fungi were unable to grow and associate with P. tabulaeformis seedlings. Inocula placed alongside O. davidiana seedlings, however, resulted in enhanced growth and nutritional status of O. davidiana and P. tabulaeformis seedlings, and also altered rhizosphere pH and phosphatase activity. We speculate that these species form a common mycorrhizal network and this association enhances outplanting performance of P. tabulaeformis seedlings used for forest restoration.     

丛枝菌根观察与侵染率测定方法的比较

菌根生长状况观察与侵染率测定是菌根学研究中一项重要的基础性工作。综述了丛枝菌根（AM）染色观察与侵染率测定方法研究概况,并对其进行比较和评价。认为采用醋酸墨水染色观察AM生长状况与采用根段侵染率加权法和放大交叉法测定AM真菌侵染率是目前较为科学、准确、易行的方法。根据不同需要也可选择其他适宜的方法,如要了解丛枝发育状况,可采用放大交叉法;如要了解泡囊和侵入点数量,可采用直接计数法,从而使其研究结果具有可比性。有必要建立基于分子生物学技术和脂肪酸定量分析技术测定一种或数种AM真菌侵染状况,这将有力推动AM真菌生理、生态功能研究的发展。     

自然入侵条件下黄顶菊丛枝菌根定殖及发育的研究

为探讨丛枝菌根(arbuscular mycorrhiza,AM)真菌在黄顶菊Flaveria bidentis入侵过程中可能发挥的作用,首先调查和研究了黄顶菊AM的侵入和生长表现。在黄顶菊入侵严重的河北省选择采样地点,分别从衡水地区的滨湖新区小北田村、冀州市漳下村、枣强县北王庄村、桃城区八里庄村、滨湖新区顺民庄村和滨湖新区刘家台村6个地点按黄顶菊重、中、轻3种盖度和不同生育期(苗期、花期和结籽期)采集根系和根区土样。分离根区土壤中AM真菌孢子、观察AM发育特征、测定孢子密度、AM真菌侵染率、丛枝着生率等,并与土壤理化性质进行了相关性分析。结果在各采样地点均观察到AM典型结构,不同样地菌丝侵染率、I型丛枝密度以及孢子密度的最大值均出现在八里庄村,泡囊密度和A型丛枝密度的最大值分别出现在小北田村和刘家台村;不同盖度下AM真菌侵染率最高值均出现在重度侵染区;不同生长时期黄顶菊的AM也有差异,除了丛枝密度最高值出现在花期和菌丝侵染率差异不显著外,菌丝侵染率、泡囊密度和孢子密度均在结籽期出现最大值。土壤理化特性也显著影响黄顶菊AM的发育,菌丝侵染率与土壤有机质呈显著正相关,与p H值呈显著负相关,与速效P含量呈极显著负相关;A型丛枝密度与p H含量呈显著负相关;I型丛枝密度与土壤全N含量呈极显著正相关;孢子密度与有机质含量和全N含量均呈显著正相关。因此,AM真菌侵染可能会促进黄顶菊的入侵。     

Receptiveness of forest soils to ectomycorrhizal association:

 Soil receptiveness to a mycorrhizal association can be estimated by standard bioassay from a dose-response relationship. The method was developed using the association Pinus pinaster or Pseudotsuga menziesii with Laccaria bicolor as a model and was successfully used to characterize the receptiveness of two forest soils. From a physical and chemical point of view, both soils were receptive to the Laccaria bicolor association. Our results show that microbial factors are very important in the receptiveness of soil to ectomycorrhizal association. Ectomycorrhizal development on seedlings at outplanting sites is discussed in relation to soil receptiveness and the ecological competence of selected strains. Accepted: 11 October 1996     

Response of ectomycorrhizal fungi to benomyl and nocodazole: growth inhibition and microtubule depolymerization

The growth of seven ectomycorrhizal fungi was tested in the presence of the antimicrotubule drugs benomyl and nocodazole. The polymerization stage of the cytoplasmic microtubules in the hyphal cells was visualized by indirect immunofluorescence microscopy after a 3-h drug treatment. Nocodazole reduced the growth of all the fungi tested at concentrations of 2 and 4 g ml^-1 and caused strong depolymerization of microtubules in all other species except Hebeloma cylindrosporum. Benomyl inhibited the growth and depolymerized the microtubules in the ascomycete Cenococcum geophilum, while in the basidiomycetes it reduced the growth and depolymerized the microtubules only in H. cylindrosporum. The role of the microtubule cytoskeleton and the target of the benzimidazole-derived drugs in fungal cells are discussed.     

Arbuscular mycorrhizal fungi in the tree seedlings of two Australian rain forests: occurrence, colonization, and relationships with plant performance

The roots of rain forest plants are frequently colonized by arbuscular mycorrhizal fungi (AMF) that can promote plant growth in the nutrient poor soils characteristic of these forests. However, recent studies suggest that both the occurrence of AMF on rain forest plants and the dependence of rain forest plants on AMF can be highly variable. We examined the occurrence and levels of AMF colonization of some common seedling species in a tropical and a subtropical rain forest site in Queensland, Australia. We also used a long-term database to compare the growth and mortality rates of seedling species that rarely formed AMF with those that regularly formed AMF. In both forests, more than one-third of the seedling species rarely formed AMF associations, while 40% of species consistently formed AMF in the tropical site compared to 27% in the subtropical site. Consistent patterns of AMF occurrence were observed among plant families at the two sites. Variation among seedling species in AMF occurrence or colonization was not associated with differences in seed mass among species, variation in seedling size and putative age within a species, or lack of AMF inoculum in the soil. Comparisons of four seedling species growing both in the shaded understory and in small canopy gaps revealed an increase in AMF colonization in two of the four species in gaps, suggesting that light limitation partially explains the low occurrence of AMF. Seedling survival was significantly positively associated with seed biomass but not with AMF colonization. Furthermore, seedling species that regularly formed AMF and those that did not had similar rates of growth and survival, suggesting that mycorrhizal and nonmycorrhizal strategies were equivalent in these forests. Furthermore, the high numbers of seedlings that lacked AMF and the overall low rate of seedling growth (the average seedling required 6 years to double its height) suggest that most seedlings did not receive significant indirect benefits from AMF through connection to canopy trees via a common mycorrhizal network.     

In situ and in vitro colonization of Cathaya argyrophylla (Pinaceae) by ectomycorrhizal fungi

Cathaya argyrophylla, a critically endangered conifer, is found to grow at four isolated areas located in subtropical mountains of China. To examine the involvement and usefulness of mycorrhizas for sustaining the population of this tree, we compared the root system, morphology, and structure of mycorrhizal roots of C. argyrophylla, which were collected from a natural stand and an artificial stand, each grown at a different location. More mycorrhizal roots were found for trees from an artificial stand. The presence of extramatrical mycelium, mantle, and Hartig net revealed that C. argyrophylla formed an ectomycorrhizal association in both sampling sites. Starch granules were found in mycorrhizal roots collected only from a natural stand. The aseptic synthesis of C. argyrophylla and Cenococcum geophilum was established for the first time in vitro. Typical ectomycorrhizas formed on seedlings on RM medium containing 0.1 g/l glucose, 5 weeks after inoculation. By light microscopy, the synthesized mycorrhizas showed a thin mantle from which emanated extramatrical hyphae and highly branched Hartig net. A simple, rapid, and convenient mycorrhiza synthesis system was developed, which facilitates further studies on ectomycorrhizal development of C. argyrophylla.     

Effects of imidazole-4-carboxamide and 2-azahypoxanthine on the growth and ectomycorrhizal colonization of Pinus densiflora seedlings inoculated with Tricholoma matsutake

Imidazole-4-carboxamide (ICA) and 2-azahypoxanthine (AHX) obtained from Lepista sordida inhibit and promote the growth of herbaceous plants, respectively. In this study, we examined the effects of these compounds on the growth and ectomycorrhizal (EM) colonization of Pinus densiflora seedlings inoculated with Tricholoma matsutake that forms EM associations with pines. The EM colonization by T. matsutake was observed on the root systems of P. densiflora seedlings treated with and without ICA and AHX. The growth of both non-EM and EM P. densiflora seedlings was inhibited by ICA, regardless of the EM colonization. In contrast, AHX promoted the growth of non-EM P. densiflora seedlings, but not of EM seedlings, suggesting that EM colonization interferes with the effect of AHX on P. densiflora growth.     

Seasonal changes of arbuscular mycorrhizal fungi as affected by tillage practices and fertilization : Hyphal density and mycorrhizal root colonization

The influence of tillage practices on native arbuscular mycorrhizal fungi (AMF) was studied in two, consecutive years in eastern Canada, in two 11 year-old long-term tillage-fertilizer experimental field soils, a sandy loam and a clay, growing corn in monoculture. The three tillage practices were: 1) conventional tillage (CT; fall plowing plus spring disking), reduced tillage (RT; spring disking) and no-till (NT). The corn crop received either inorganic (N and K) or organic (liquid dairy manure) fertilizers. Mycorrhizal hyphal density was estimated from soil samples obtained in early spring (before disking), at the 12–14 leaf stage, at silking, and at harvest. The percentage of corn root colonization by AMF at the 12–14 leaf stage, at silking and at harvest was also determined. The sandy loam was sampled over two consecutive seasons and the clay soil over one season.Densities of total and metabolically active soil hyphae, and mycorrhizal root colonization were significantly lower in CT soil than in RT and NT soil. Lowest soil hyphal densities were observed in early spring. The levels of intra- and extraradical fungal colonization always increased from spring to silking and decreased thereafter. Spring disking had only a small and transient negative effect on hyphal abundance in soil. Fertilization did not influence mycorrhizal colonization of corn or abundance of soil hyphae in the sandy loam soil, but in the clay soil metabolically active hyphae were more abundant with manure application than with mineral fertilization. In 1992, in both soils different tillage systems had same grain yield, however, in 1993, corn yield was higher in NT compared to CT system.     

Community structure of ectomycorrhizal fungi in a Pinus muricata forest: minimal overlap between the mature forest and resistant propagule communities

We have investigated colonization strategies by comparing the abundance and frequency of ectomycorrhizal fungal species on roots in a mature Pinus muricata forest with those present as resistant propagules colonizing potted seedlings grown in the same soil samples. Thirty-seven fungal species were distinguished by internal transcribed spacer (ITS) restriction fragment length polymorphisms (RFLPs); most were identified to species level by sporocarp RFLP matches or to genus/family level by using sequence databases for the mitochondrial and nuclear large-subunit rRNA genes. The below-ground fungal community found in the mature forest contrasted markedly with the resistant propagule community, as only four species were found in both communities. The dominant species in the mature forest were members of the Russulaceae, Thelephorales and Amanitaceae. In contrast, the resistant propagule community was dominated by Rhizopogon species and by species of the Ascomycota. Only one species, Tomentella sublilacina (Thelephorales), was common in both communities. The spatial distribution of mycorrhizae on mature roots and propagules in the soil differed among the dominant species. For example, T. sublilacina mycorrhizae exhibited a unique bias toward the organic horizons, Russula brevipes mycorrhizae were denser and more clumped than those of other species and Cenococcum propagules were localized, whereas R. subcaerulescens propagules were evenly distributed. We suggest that species differences in resource preferences and colonization strategies, such as those documented here, contribute to the maintenance of species richness in the ectomycorrhizal community.     

Rapid nitrogen transfer from ectomycorrhizal pines to adjacent ectomycorrhizal and arbuscular mycorrhizal plants in a California oak woodland

Nitrogen transfer among plants in a California oak woodland was examined in a pulse-labeling study using 15N. The study was designed to examine N movement among plants that were mycorrhizal with ectomycorrhizas (EM), arbuscular mycorrhizas (AM), or both. Isotopically enriched N (K15NO3-) was applied to gray pine (Pinus sabiniana) foliage (donor) and traced to neighboring gray pine, blue oak (Quercus douglasii), buckbrush (Ceanothus cuneatus) and herbaceous annuals (Cynosurus echinatus, Torilis arvensis and Trifolium hirtum). After 2 wk, needles of 15N-treated pines and foliage from nearby annuals were similarly enriched, but little 15N had appeared in nontreated (receiver) pine needles, oak leaves or buckbrush foliage. After 4 wk foliar and root samples from pine, oak, buckbrush and annuals were significantly 15N-enriched, regardless of the type of mycorrhizal association. The rate of transfer during the first and second 2-wk periods was similar, and suggests that 15N could continue to be mobilized over longer times.     

Effects of manipulation of litter and humus layers on ectomycorrhizal colonization potential in Scots pine stands of different age

Effects of manipulation of litter and humus layers (removal, doubling and control treatments) on the colonization potential of ectomycorrhizal fungi were studied in two secondary stands of Pinus sylvestris (5 and 18 years old) in The Netherlands. Five-mont-hold, sterile-grown Scots pine seedlings, inoculated with Laccaria bicolor, Paxillus involutus or Rhizopogon luteolus and noninoculated seedlings were used as baits. The seedlings were harvested after one growing season. For comparison, sporocarps of ectomycorrhizal fungi were also investigated. Genus composition on the seedlings was independent of initial inoculum, but determined by both treatment and age of the stands. In both stands, removal of litter and humus layers increased, and addition of organic material decreased the number of ectomycorrhizal types on the seedlings. Not all indigenous genera were observed by either outplanting seedlings or sporocarp surveys.     

Application of network theory to potential mycorrhizal networks

The concept of a common mycorrhizal network implies that the arrangement of plants and mycorrhizal fungi in a community shares properties with other networks. A network is a system of nodes connected by links. Here we apply network theory to mycorrhizas to determine whether the architecture of a potential common mycorrhizal network is random or scale-free. We analyzed mycorrhizal data from an oak woodland from two perspectives: the phytocentric view using trees as nodes and fungi as links and the mycocentric view using fungi as nodes and trees as links. From the phytocentric perspective, the distribution of potential mycorrhizal links, as measured by the number of ectomycorrhizal morphotypes on trees of Quercus garryana, was random with a short tail, implying that all the individuals of this species are more or less equal in linking to fungi in a potential network. From the mycocentric perspective, however, the distribution of plant links to fungi was scale-free, suggesting that certain fungus species may act as hubs with frequent connections to the network. Parallels exist between social networks and mycorrhizas that suggest future lines of study on mycorrhizal networks.     

Niche breadth,environmental landscape,and physical barriers: their importance as determinants of species distributions

Species distributions and their patterns in geographical space have been studied for several decades and explained by theories such as Janzen's, with respect to the nature of dispersal barriers in the Tropics, and Rapoport's, with respect to range size. However, the roles of specific environmental and geographical factors (e.g. ecological niche breadth, geographical barriers, etc.) in shaping species ranges and distributional patterns remain largely unexplored. The present study analyzed predictions from these two theories via analysis of virtual species with respect to biogeographical patterns: virtual species were created across South America, covering all major environments on the continent, and were used to compare effects of niche breadth, environmental availability, connectivity, seasonality, and the presence of known biogeographical barriers (rivers) in shaping species distributions and biodiversity patterns. Geographical ranges varied from narrow to broad, depending on the location of the seed point when comparing species produced with the same niche breadth. Analysis without consideration of seasonality and barriers produced species with broader distributions in the Tropics and narrower distributions in montane and temperate regions of the continent. When seasonality was included, however, broader ranges were concentrated in temperate regions, thus supporting Janzen's idea. Rapoport's rule of broader geographical ranges at higher latitudes was supported only when seasonality and physical barriers were included but not in species with very narrow or very broad niches, suggesting that this ‘rule’ results from interactions among niche breadth, dispersal capabilities, and dispersal barriers. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 108 , 241–250.