Effect of long-term nitrogen fertilization on mycorrhizal fungi associated with a dominant grass in a semiarid grassland

We studied the diversity of arbuscular mycorrhizal fungi (AMF) in semiarid grassland and the effect of long-term nitrogen (N) fertilization on this fungal community. Root samples of Bouteloua gracilis were collected at the Sevilleta National Wildlife Refuge (New Mexico, USA) from control and N-amended plots that have been fertilized since 1995. Small subunit rDNA was amplified using AMF specific primers NS31 and AM1. The diversity of AMF was low in comparison with other ecosystems, only seven operational taxonomic units (OTU) were found in B. gracilis and all belong to the genus Glomus. The dominant OTU was closely related to the ubiquitous G. intraradices/G. fasciculatum group. N-amended plots showed a reduction in the abundance of the dominant OTU and an increase in AMF diversity. The greater AMF diversity in roots from N-amended plots may have been the result of displacement of the dominant OTU, which facilitated detection of uncommon AMF. The long-term implications of AMF responses to N enrichment for plant carbon allocation and plant community structure remain unclear.     

Flavonoids in roots of white clover: interaction of arbuscular mycorrhizal fungi and a pathogenic fungus

The effects of two arbuscular mycorrhizal fungi (AMF) (Glomus mosseae and G. claroideum) and a pathogenic fungus (Pythium ultimum) on the production of eight flavonoids in roots of two white clover (Trifolium repens L.) cultivars were evaluated. Quantification of AM and pathogenic fungi in the roots showed that the AM symbiosis significantly reduced P. ultimum biomass and in some cases prevented infection. The flavonoid productions in clover roots varied depending on the presence of beneficial and/or pathogenic fungi, fungal isolate or plant cultivar. Only plants colonized with G. claroideum showed detectable concentrations of either coumestrol or kaempferol (cultivar-dependant). In addition, inoculation with G. claroideum resulted in significantly higher concentrations of coumestrol in cv. Sonja and medicarpin in cv. Milo. A low production of coumestrol and kaempferol in mycorrhizal plants may be G. mosseae-specific. Only the concentrations of formononetin and daidzein increased in clover roots in response to infection with P. ultimum. These flavonoids are supposedly stress metabolites, synthesized or produced from glycosides in response to pathogen infection. However, the presence of one or both AMF significantly lowered the formononetin and daidzein concentrations, and overruled the inductive effect of P. ultimum. Therefore the antagonistic action of AM against the pathogen must take place through another mechanism.     

PCR with degenerate primers amplifies a subgenomic DNA fragment from the endoglucanase gene(s) of Torula thermophila, a thermophilic fungus

The aim of this study was to enable the polymerase chain reaction (PCR) amplification of DNA fragments within endoglucanase gene(s) of Torula thermophila, by using degenerate primers so that the amplified fragment(s) could be used as homologous probe(s) for cloning of full-length endoglucanase gene(s). The design of the degenerate PCR primers was mainly based on the endoglucanase sequences of other fungi. The endoglucanase gene sequence of Humicola insolens was the only sequence from a thermophilic fungus publicly available in the literature. Therefore, the endoglucanase sequences of the two Trichoderma species, Trichoderma reesei and Trichoderma longibrachiatum, were used to generalize the primers. PCR amplification of T. thermophila genomic DNA with these primers resilied in a specific amplification. The specificity of the amplified fragment was shown by Southern hybridization analysis using egl3 gene of T. reesei as probe. This result suggested that the degenerate primers used in this study may be of value for studies aimed at cloning of endoglucanase genes from a range of related fungi.     

Defence-related Enzymes in Pepper Roots During Interactions with Arbuscular Mycorrhizal Fungi and/or Verticillium dahliae

Previous studies have described that arbuscular mycorrhizal fungi (AMF) can reduce the deleterious effect of Verticillium dahliae Kleb. on pepper growth and yield. In mycorrhizal plants, the bioprotection against soil-borne pathogens can result from the preactivation of defence responses that include some structural modifications and the accumulation of Pathogenesis-Related (PR) proteins. Our first objective was to study if V. dahliae induced defence mechanisms in roots before infected pepper developed visible symptoms of disease. The second aim was to determine if AMF induced defence-related enzymatic activities in pepper roots before or after pathogen’s attack. Results showed that the colonization of pepper roots by Glomus deserticola (Trappe, Bloss and Menge) induced the appearance of new isoforms of acidic chitinases, superoxide dismutase (SOD) and, at early stages, peroxidases. In contrast, V. dahliae neither stimulated the phenylpropanoid pathway nor elicited hydrolytic activities in infected pepper roots. Only in mycorrhizal plants, the inoculation with V. dahliae slightly increased both phenylalanine ammonia-lyase (PAL) and peroxidase activities two weeks later. Mycorrhizal-specific induction of new isoforms of acidic chitinases and SOD together with enhanced peroxidase and PAL activities 2 weeks after pathogen inoculation could be involved in the biocontrol of Verticillium-induced wilt in pepper by AMF.     

Fern-associated arbuscular mycorrhizal fungi are represented by multiple Glomus spp.: do environmental factors influence partner identity?

Symbioses involving arbuscular mycorrhizal fungi (AMF) are among the most important ecological associations for many plant species. The diversity of AMF associated with ferns, however, remains poorly studied. Using recently designed Glomus-specific primers, we surveyed the AMF community associated with ferns from deciduous, broad-leaved second-growth forest habitats at the eastern edge of the piedmont region of central Virginia, USA. Results indicate that this molecular approach may be a useful tool for detecting AMF in ferns compared to traditional techniques based on morphology. Over 30 potential fungal ribotypes were identified from eight fern species using denaturing gradient gel electrophoresis. Fungal ribotypes were found to differ widely in terms of (1) the number of fern partners with which they interact and (2) their relative frequency within each fern. Sequence analysis of fungal isolates from three species of fern indicated that the primers were generally highly specific for Glomus species but some non-target DNA was also amplified. Cloned polymerase chain reaction (PCR) products from Polystichum acrostichoides and Osmunda regalis revealed several phylogenetically distinct Glomus species. A single Glomus species was identified in the cloned PCR products from Botrychium virginianum. These findings challenge the hypothesis that the extent or degree of fern–fungal symbiosis is somehow tied to root complexity. Environmental factors appear to influence the suite of AMF that form partnerships with ferns. Some species of fern from similar habitats associated with dissimilar fungal partners (e.g., P. acrostichoides and Athyrium filix-femina var. asplenioides), whereas others harbored uniform fungal communities (e.g., Asplenium platyneuron). The significance of these data in terms of ecological and evolutionary dynamics of the AMF–fern symbiosis is discussed. Brittany West, Jessica Brandt, and Kay Holstien contributed equally to this work.     

Simulated nitrogen deposition affects community structure of arbuscular mycorrhizal fungi in northern hardwood forests

Our previous investigation found elevated nitrogen deposition caused declines in abundance of arbuscular mycorrhizal fungi (AMF) associated with forest trees, but little is known about how nitrogen affects the AMF community composition and structure within forest ecosystems. We hypothesized that N deposition would lead to significant changes in the AMF community structure. We studied the diversity and community structure of AMF in northern hardwood forests after more than 12 years of simulated nitrogen deposition. We performed molecular analyses on maple (Acer spp.) roots targeting the 18S rDNA region using the fungal‐specific primers AM1 and NS31. PCR products were cloned and identified using restriction fragment length polymorphism (RFLP) and sequencing. N addition significantly altered the AMF community structure, and Glomus group A dominated the AMF community. Some Glomus operational taxonomic units (OTUs) responded negatively to N inputs, whereas other Glomus OTUs and an Acaulospora OTU responded positively to N inputs. The observed effect on community structure implies that AMF species associated with maples differ in their response to elevated nitrogen. Given that functional diversity exists among AMF species and that N deposition has been shown to select less beneficial fungi in some ecosystems, this change in community structure could have implications for the functioning of this type of ecosystem.     

Survival of inoculated Laccaria bicolor in competition with native ectomycorrhizal fungi and effects on the growth of outplanted Douglasfir seedlings

The survival, development and mycorrhizal efficiency of a selected strain of Laccaria bicolor along with naturally occurring ectomycorrhizal fungi in a young plantation of Douglas fir was examined. Symbionts were identified and their respective colonization abilities were determined. Eight species of symbiotic fungi, which may have originated in adjacent coniferous forests, were observed on the root systems. Mycorrhizal diversity differed between inoculated (5 taxa) and control (8 taxa) seedlings. Ectomycorrhizal fungi which occurred naturally in the nursery on control seedlings (Thelephora terrestris and Suillus sp.) did not survive after outplanting. Both inoculated and naturally occurring Laccaria species, as well as Cenococcum geophilum, survived on the old roots and colonized the newly formed roots, limiting the colonization by other naturally occurring fungi. Other fungi, such as Paxillus involutus, Scleroderma citrinum and Hebeloma sp. preferentially colonized the old roots near the seedling's collar. Russulaceae were found mainly in the middle section of the root system. Mycorrhizal colonization by Laccaria species on inoculated seedlings (54%) was significantly greater than on controls (13%) which were consequently dominated by the native fungi. Significant differences (up to 239%) were found in the growth of inoculated seedlings, especially in root and shoot weight, which developed mainly during the second year after outplanting. Seedling growth varied with the species of mycorrhizae and with the degree of root colonization. Competitiveness and effectiveness of the introduced strain on improving growth performances of seedlings are discussed.     

Neighboring plant influences on arbuscular mycorrhizal fungal community composition as assessed by T-RFLP analysis

Controls on root colonization by arbuscular mycorrhizal fungi (AMF) include host nutrient status, identity of symbionts and soil physico-chemical properties. Here we show, in the field, that the subset of the AMF community colonizing the roots of a common grass species, Dactylis glomerata, was strongly controlled by neighboring roots of a different plant species, Centaurea maculosa, an invasive forb, thus adding a biological spatial component to controls on root colonization. Using an AMF-specific, 18s rDNA-based terminal restriction fragment length polymorphism (T-RFLP) analysis method, significant differences were found between AMF community fingerprints of samples derived from roots of grasses with (G_Cm) and without (G₀) neighboring C. maculosa. There were also significant differences between samples derived from C. maculosa roots (C_mac) and both G_Cm and G₀ roots. Sample ordination indicated three generally distinct groups consisting of C_mac, G_Cm and G₀, with G_Cm samples being of intermediate distance between G₀and C_mac. Our results indicate that, with the presence of C. maculosa, AMF communities of D. glomerata shift to reflect community composition associated with C. maculosa roots. These results highlight the importance of complex spatial distributions of AMF communities at the scale of a root system. An additional dimension to our study is that C. maculosa is an aggressively invasive plant in the intermountain West. Viewed in this light, these results suggest that pervasive influences of this plant on AMF communities, specifically in roots of its competitors, may represent a mechanism contributing to its invasive success. However, further work is clearly required to determine the extent to which AMF genotypic alteration by neighboring plants influences competitive relationships.     

连作花魔芋软腐病株与健株根域丛枝菌根真菌群落多样性

【目的】解析不同连作年限花魔芋软腐病株、健株根域的丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)群落多样性。【方法】使用AMF 18S SSU rRNA基因特异引物AMV4.5NF/AMDGR对正茬及连作2年和3年的软腐病株、健株魔芋根系和根际土壤DNA扩增建库,通过高通量测序和生物信息学分析探究魔芋软腐病与其根域AMF群落多样性的关系。【结果】魔芋根系具有明显的AMF菌丝、泡囊和丛枝等结构。在相同连作年限条件下,健株根系AMF总侵染率、侵染强度和孢子密度均显著高于病株(P<0.05);在不同连作年限条件下,病株根系AMF总侵染率和侵染强度随连作年限延长而降低。从所有样品中共鉴定到9属53种AMF,其中有49个已知种和4个新种。球囊霉属(Glomus)和类球囊霉属(Claroideoglomus)是AMF群落的优势属,其AMF种分别占总AMF种数的41.5%和26.4%;丰度最高的Paraglomus sp.VTX00308是所有样品的共有种。连作、软腐病及二者的交互作用显著影响根系AMF群落的Shannon指数和Simpson指数及根际土壤AMF的Chao1指数(P<0.05)。通过丰度差异分析发现6个在连作软腐病发生后丰度差异显著的AMF种(P<0.05);NMDS分析表明,不同连作年限的魔芋软腐病株与健株之间的根域AMF菌种组成、相对丰度和群落结构存在差异。相关性分析表明,软腐病发病率和病情指数与魔芋根系和根际土壤AMF的Shannon指数、根系AMF的Chao1和Simpson指数以及AMF总侵染率、侵染强度和孢子密度极显著负相关(P<0.01)。【结论】比对健株,连作魔芋软腐病株根际土壤AMF孢子密度以及根系AMF侵染率、种数和多样性均降低,其群落结构显著改变。     

Presence of arbuscular mycorrhizal fungi in South Florida native plants

The roots of 27 species of South Florida plants in 15 families (including one cycad, six palms, one Smilax, and 19 dicotyledons) native to pine rockland and tropical hardwood hammock communities were examined for arbuscular mycorrhizal fungi (AMF). These plants grow in the biologically diverse but endangered Greater Everglades habitat. Roots from field-grown and potted plants were cleared and stained. All 27 species had AMF and include 14 species having an endangered or threatened status. The Paris-type colonization occurred in two species in the families Annonaceae and Smilacaceae. The Arum-type occurred in 22 species in the families Anacardiaceae, Arecaceae (Palmae), Boraginaceae, Cactaceae (questionable), Euphorbiaceae, Fabaceae, Lauraceae, Melastomataceae, Polygalaceae, Rubiaceae, Simaroubaceae, Ulmaceae, and Zamiaceae. Three species in the families Fabaceae, Lauraceae, and Simaroubaceae had a mix of Paris- and Arum-types. The results have implications for the restoration of these endangered plant communities in the Everglades.     

Nested multiplex PCR—A feasible technique to study partial community of arbuscular mycorrhizal fungi in field-growing plant root

Plant can be infected by different arbuscular mycorrhizal fungi, but little is known about the interaction between them within root tissues mainly because different species cannot be distinguished on the basis of fungal structure. Accurate species identification of Arbuscular mycorrhizal fungi (AMF) colonized in plant roots is the comerstone of mycorrhizal study, yet this fundamental step is impossible through its morphological character alone. For accurate, rapid and inexpensive detection of partial mycorrhizal fungal community in plant roots, a nested multiplex polymerase chain reaction (PCR) was developed in this study. Five discriminating primers designed based on the variable region of the 5′ end of the large ribosomal subunit were used in the experiment for testing their specificity and the sensitivity in nested PCR by using spores from Glomus mosseae (BEG12), Glomus intraradices (BEG141), Scutellospora castaneae (BEG1) and two unidentified Glomus sp. HAUO3 and HAUO4. The feasibility assay of nested multiplex PCR was conducted by use of spore mixture, Astragalus sinicum roots co-inoculated with 4 species of arbuscular mycorrhizal fungi from pot cultures and 15 different field-growing plant roots respectively after analyses of the compatibility of primers. The result indicated that the sensitivity was in the same range as that of the corresponding single PCR reaction. Overall accuracy was 95%. The efficiency and sensitivity of this multiplex PCR procedure provided a rapid and easy way to simultaneously detect several of arbuscular mycorrhizal fungal species in a same plant root system.     

干旱胁迫下AMF对云南蓝果树幼苗生长和光合特征的影响

采用盆栽试验与称重控水法,将土壤相对含水量分别控制在田间最大持水量的100%、91.68%、82.85%、60.00%、41.86%和21.28%,并在这6个不同的土壤相对含水量条件下,分别设添加苯菌灵(杀真菌剂)(低AMF)和不添加苯菌灵(高AMF)处理,研究干旱胁迫下AMF对极小种群野生植物云南蓝果树幼苗生长和光合特征的影响,揭示云南蓝果树濒危的微生物学机制,为云南蓝果树保护措施的制定与实施奠定基础。结果表明,添加苯菌灵处理显著降低了不同水分条件下的AMF侵染率,说明试验中AMF处理的实生苗在生长和光合特征上的差异是苯菌灵处理下侵染率下降导致的;随着干旱胁迫的加剧,云南蓝果树幼苗的根部AMF侵染率显著降低、叶面积等生长指标和净光合速率(Pn)等光合参数都发生显著变化;高AMF处理可以显著增加水分充足和轻度干旱胁迫条件下云南蓝果树幼苗的大部分生长指标和光合参数,而对重度胁迫下的云南蓝果树幼苗没有显著影响,说明重度干旱胁迫对其影响大于AMF的影响;另外,整合了可塑性指数分析和隶属函数分析两种方法对其抗旱性进行评价,云南蓝果树幼苗基本上无法通过调节形态和光合能力来适应水分环境的变化,但是高AMF处理可使云南蓝果树幼苗具有较强的可塑性和更强的抗旱性。实验结果为云南蓝果树的科学保育及种苗繁育提供了理论依据。     

Detection of the ectomycorrhizal fungusTricholoma matsutake and some related species with specific ITS primers

Oligonucleotide primers (Tm1 and Tm4) were designed to amplify a 447–448 base pair fragment, comprising sections of the rDNA internal transcribed spacers (ITS) and the entire 5.8S rDNA, ofTricholoma matsutake. PCR products of predicted size were produced for six of eight isolates ofT. matsutake from across its natural range in Asia, and for isolates of some closely related fungi includingT. bakamatsutake, T. magnivelare, andT. caligatum. The closely relatedT. robustum could be discriminated fromT. matsutake by PCR fragment size. No PCR products were produced where the primers were tested against 16 species of ectomycorrhizal fungi associated withPinus spp. in the Southern Hemisphere. The specific primers were also used successfully to produce PCR products from matsutake infected roots collected in natural forests in China and Japan, and from pure culture synthesisedPinus radiata-T. matsutake material. These primers will be useful in research directed at establishing matsutake in the Southern Hemisphere, and also have the potential to be applied to the study of matsutake within its natural range.     

白及内生真菌多样性研究

白及( Bletilla striata)是兰科地生型多年生植物,也是我国传统中药材之一。利用菌根技术进行白及的保护和人工栽培,需要获得白及可培养的内生真菌。该研究以广西野生的白及根和叶为材料,采用分离培养法分离内生真菌,并结合真菌形态特征,及其核糖体的转录间隔区( ITS)序列分析,确定内生真菌的分类地位。结果表明：从2株白及植物90块组织中分离获得37株内生真菌,鉴定为15个分类单元,由9个属组成,分属于2门4纲7目8科,包括锤舌菌纲( Leotiomycetes)、座囊菌纲( Dothideomycetes)和粪壳菌纲( Sordariomy-cetes),伞菌纲( Agaricomycetes)。从根中分离获得内生真菌12种,蜡壳菌属为优势属;从叶中分离获得内生真菌3种,刺盘孢属为优势属;刺盘孢菌属( Colletotrichum)和蜡壳菌属( Sebacina)真菌的相对多度值均达到20％;4株担子菌均分布于根中,叶组织中未有分布。根组织中内生真菌的多样性指数(H＝1.863)高于叶组织(1.098)。该研究结果及其所分离培养的担子菌类真菌,为更好地利用菌根技术进行白及等兰科植物资源的保护与可持续利用奠定了基础。     

Nested multiplex PCR——A feasible technique to study partial community of arbuscular mycorrhizal fungi in field-growing plant root

Plant can be infected by different arbuscular mycorrhizal fungi, but little is known about the interaction between them within root tissues mainly because different species cannot be distinguished on the basis of fungal structure. Accurate species identification of Arbuscular mycorrhizal fungi (AMF) colonized in plant roots is the comerstone of mycorrhizal study, yet this fundamental step is impossible through its morphological character alone. For accurate, rapid and inexpensive detection of partial mycorrhizal fungal community in plant roots, a nested multiplex polymerase chain reaction (PCR) was developed in this study. Five discriminating primers designed based on the variable region of the 5′ end of the large ribosomal subunit were used in the experiment for testing their specificity and the sensitivity in nested PCR by using spores from Glomus mosseae (BEG12), Glomus intraradices (BEG141), Scutellospora castaneae (BEG1) and two unidentified Glomus sp. HAUO3 and HAUO4. The feasibility assay of nested multiplex PCR was conducted by use of spore mixture, Astragalus sinicum roots co-inoculated with 4 species of arbuscular mycorrhizal fungi from pot cultures and 15 different field-growing plant roots respectively after analyses of the compatibility of primers. The result indicated that the sensitivity was in the same range as that of the corresponding single PCR reaction. Overall accuracy was 95%. The efficiency and sensitivity of this multiplex PCR procedure provided a rapid and easy way to simultaneously detect several of arbuscular mycorrhizal fungal species in a same plant root system.     

Molecular diversity of arbuscular mycorrhizal fungi in onion roots from organic and conventional farming systems in the Netherlands

Diversity and colonization levels of naturally occurring arbuscular mycorrhizal fungi (AMF) in onion roots were studied to compare organic and conventional farming systems in the Netherlands. In 2004, 20 onion fields were sampled in a balanced survey between farming systems and between two regions, namely, Zeeland and Flevoland. In 2005, nine conventional and ten organic fields were additionally surveyed in Flevoland. AMF phylotypes were identified by rDNA sequencing. All plants were colonized, with 60% for arbuscular colonization and 84% for hyphal colonization as grand means. In Zeeland, onion roots from organic fields had higher fractional colonization levels than those from conventional fields. Onion yields in conventional farming were positively correlated with colonization level. Overall, 14 AMF phylotypes were identified. The number of phylotypes per field ranged from one to six. Two phylotypes associated with the Glomus mosseae–coronatum and the G. caledonium–geosporum species complexes were the most abundant, whereas other phylotypes were infrequently found. Organic and conventional farming systems had similar number of phylotypes per field and Shannon diversity indices. A few organic and conventional fields had larger number of phylotypes, including phylotypes associated with the genera Glomus-B, Archaeospora, and Paraglomus. This suggests that farming systems as such did not influence AMF diversity, but rather specific environmental conditions or agricultural practices. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Authors Guillermo A. Galván and István Parádi contributed equally to this research and share first co-authorship.     

干旱条件下接种AM真菌对小马鞍羊蹄甲幼苗根系的影响

为了探讨岷江干旱河谷丛枝菌根真菌(AMF)对寄主植物幼苗根系的影响,通过接种购买的AMF摩西球囊霉菌(Funneliformis mosseae)到优势乡土灌木小马鞍羊蹄甲(Bauhinia faberi var.microphylla)幼苗,在重度、中度和轻度干旱条件下培养3个月,研究不同干旱条件下AMF对幼苗根系形态特征、结构特征、功能性状的影响。方差分析结果表明:(1)3种干旱胁迫条件下,接菌均显著增加了幼苗的根总长、根表面积、根分枝数、根尖数(P0.001),在中度胁迫和轻度胁迫下,接菌显著促进根鲜重、根体积的增加(P0.001),轻度胁迫条件下接菌幼苗的根鲜重、根总长、根表面积、根体积、根尖数最高并显著高于其它处理,但接菌与未接菌的根平均直径之间没有显著差异;(2)接菌幼苗根系趋向于叉状分支结构,在重度胁迫时,叉状分支趋势更显著(P0.001);(3)接菌幼苗的根比例都显著小于未接菌的,但幼苗比根长不存在显著差异。相关分析结果表明:菌根侵染率与根鲜重、根总长、根表面积、根体积、根分枝数、根尖数呈极显著正相关(P0.001),与拓扑指数、根比例呈极显著负相关(P0.001)。研究表明,在干旱条件下,AMF虽然没有提高生长初期的根系的吸收效率,但接种AMF显著影响幼苗根系形态特征和结构特征,更利于植物适应干旱环境,并且AMF对幼苗根系的促生作用随着干旱胁迫程度减轻而提高。     

Phosphorus Acquisition Strategies within Arbuscular Mycorrhizal Fungal Community of a Single Field Site

Diversity in phosphorus (P) acquisition strategies was assessed among eight isolates of arbuscular mycorrhizal fungi (AMF) belonging to three Glomus species, all obtained from the same field site. Maize (Zea mays L. cv. Corso) was used as a test plant. Compartmented cultivation containers coupled with ³³P radioisotope labeling of soil P were employed to estimate (1) the distance from the roots that AMF were able to acquire soil P from, (2) the rate of soil colonization, (3) the efficiency of uptake of soil P by AMF, (4) benefits provided to maize in terms of P acquisition and growth. Glomus mosseae and G. intraradices took up P 10 cm from roots, whereas G. claroideum only up to 6 cm from the roots. G. mosseae most rapidly colonized the available soil volume and transported significant amounts of P to maize from a distance, but provided no net P uptake benefit to the plants. On the other hand, both G. intraradices and three out of four G. claroideum isolates significantly improved net P uptake by maize. These effects seem to be related to variability between and to a limited extent also within AMF species, in mycelium development, efficiency of hyphal P uptake and effects on plant P acquisition via the root pathway. In spite of absence of maize growth responses to inoculation with any of the AMF isolates, this study indicates remarkable functional diversity in the underground component of the studied field site.     

Identification of Frankia strains in nodules by hybridization of polymerase chain reaction products with strain-specific oligonucleotide probes

A set of oligonucleotides has been developed to study the competitivity of two Frankia strains in the nodulation of the roots of two host plant species: Alnus glutinosa and Alnus incana. Two 20 mer-oligonucleotides, complementary to highly conserved sequences inside the nifH gene, were used as primers for the polymerase chain reaction (PCR) system in order to amplify microsymbiont DNA extracted from actinorhizae. PCR products were analyzed using two strain-specific 15-mer oligonucleotides identified in the amplified region. Hybridization data indicate that strain ACoN24d is more competitive than train ArI3 in the nodulation of both hosts.     

Multiple species of ectomycorrhizal fungi are frequently detected on individual oak root tips in a tropical cloud forest

The ecological importance of ectomycorrhizal (EM) fungi in tropical ecosystems is increasingly recognized, but few studies have used molecular methods to examine EM fungal communities in tropical forests. The diversity and composition of the EM community on Quercus crassifolia in a tropical montane cloud forest in southern Mexico were characterized using DNA sequencing of single root tips. Individual root tips commonly harbored multiple fungal species that resulted in mixed polymerase chain reaction (PCR) products. By cloning and performing gel extractions on mixed PCR samples, we identified two or more EM fungi on 26% of the root tips. When non-EM fungi were considered, this figure increased to 31% of root tips. A total of 44 EM taxa and nine non-EM taxa were detected on roots from 21 soil cores (104 root tips). Taxa in the families Russulaceae, Cortinariaceae, Inocybaceae, and Thelephoraceae were frequent. This is the first study to characterize the belowground EM community in a tropical montane cloud forest. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.