The application of green synthesis in the nano-science and technology is of great importance in the area of the preparation of various materials. In this work, three selected algal species Parachlorella kessleri, Dictyosphaerium chlorelloides and Desmodesmus quadricauda were successfully used for the preparation of silver nanoparticles (AgNPs). Presence of AgNPs was confirmed by UV-vis spectroscopy and transmission electron microscopy. AgNPs produced by P. kessleri had narrow size distribution and average sizes of 7.6 nm. However, nanoparticle production lasted for long time. Nanoparticle formation by D. chlorelloides was the fastest, although, their average sizes were 23.4 nm with broad size distribution. Nanoparticles produced by D. quadricauda had average sizes 23.9 nm but they were the least stable, aggregated and precipitated from solutions within 3 days. These results confirmed that the size distribution and mean diameter of the nanoparticles, crucial for various applications, can be controlled by the organism selection.

Genetic, phenotypic and functional variation in a Glomus geosporum isolate obtained from a highly alkaline anthropogenic sediment was investigated. Two cultivation lineages of G. geosporum (BEG199 and BEG211) were created by sub-culturing with or without the stress of the sediment. Genetic variation was assessed on spores from both cultivation lineages in the large sub-unit rRNA gene D2 region using PCR-single strand conformational polymorphism (PCR-SSCP) and sequencing. Phenotypical and functional variation of the cultivation lineages were assessed after inoculation onto Conyza bilbaoana. The sequence diversity obtained in G. geosporum BEG211 was significantly different from that obtained in G. geosporum BEG199. Glomus geosporum BEG199 was more effective than G. geosporum BEG211 in promoting the growth of C. bilbaoana in inert substrate and in increasing its leaf phosphorus concentration when the plant was grown in sediment. After inoculation onto C. bilbaoana, G. geosporum BEG199 colonised the roots to a greater extent, produced a significantly larger number of spores and presented a greater length of extraradical mycelium than G. geosporum BEG211. The results indicate that environmental conditions under which arbuscular mycorrhizal fungi (AMF) are cultivated can influence their effects in host plants. Also, AMF might quickly lose gained-tolerance to environmental stresses when maintained without the selective pressure of those stresses. This study has implications for the production of commercial inoculum of AMF and for the maintenance of AMF cultures. The results indicate that inoculum of AMF should be produced and AMF cultures should be maintained in substrates or media containing the original edaphic stresses.

The Petri-dish microcosm is a simple system to study the development of arbuscular mycorrhizal fungi (AMF) and has been shown to be suitable to observe the establishment of Acaulospora tuberculata BEG41, Gigaspora rosea BEG111, Glomus manihotis BEG112 and Scutellospora heterogama BEG40 on plant root. The non-agar based system could be easily used to observe the whole course of development of fungal-root interactions as well as the extra-radical structures of AMF without destructive sampling. It was found that these AMF species were unique with regard to spore germination, root colonization, the architecture of the extra-radical mycelium (ERM), the spread of the ERM in the media, and sporulation. The ERM of A. tuberculata BEG41and G. manihotis BEG112 spread mainly close to the root system, while the ERM of G. rosea BEG111 and S. heterogama BEG40 could occupy areas unoccupied by roots. The unique structure, the branch absorbing structure, was found in the four species of AMF, but differed in their shape and size

The ultrastructural detail of spore development in Scutellospora heterogama is described. Although the main ontogenetic events are similar to those described from light microscopy, the complexity of wall layering is greater when examined at an ultrastructural level. The basic concept of a rigid spore wall enclosing two inner, flexible walls still holds true, but there are additional zones within these three walls distinguishable using electron microscopy, including an inner layer that is involved in the formation of the germination shield. The spore wall has three layers rather than the two reported previously. An outer, thin ornamented layer and an inner, thicker layer are both derived from the hyphal wall and present at all stages of development. These layers differentiate into the outer spore layer visible at the light microscope level. A third inner layer unique to the spore develops during spore swelling and rapidly expands before contracting back to form the second wall layer visible by light microscopy. The two inner flexible walls also are more complex than light microscopy suggests. The close association with the inner flexible walls with germination shield formation consolidates the preferred use of the term 'germinal walls' for these structures. A thin electron-dense layer separates the two germinal walls and is the region in which the germination shield forms. The inner germinal wall develops at least two sub-layers, one of which has an appearance similar to that of the expanding layer of the outer spore wall. An electron-dense layer is formed on the inner surface of the inner germinal wall as the germination shield develops, and this forms the wall surrounding the germination shield as well as the germination tube. At maturity, the outer germinal wall develops a thin, striate layer within its substructure.

Different species of arbuscular mycorrhizal fungi (AMF) can produce different amounts of extraradical mycelium (ERM) with differing architectures. They also have different efficiencies in gathering phosphate from the soil. These differences in phosphate uptake and ERM length or architecture may contribute to differential growth responses of plants and this may be an important contributor to plant species coexistence. The effects of the development of the ERM of AMF on the coexistence of two co-occurring plant species were investigated in root-free hyphal chambers in a rhizobox experimental unit. The dominant shrub (Salix atrocinerea Brot.) and herbaceous (Conyza bilbaoana J. Rémy) plant species found in a highly alkaline anthropogenic sediment were studied in symbiosis with four native AMF species (Glomus intraradices BEG163, Glomus mosseae BEG198, Glomus geosporum BEG199 and Glomus claroideum BEG210) that were the most abundant members of the AMF community found in the sediment. Different AMF species did not influence total plant productivity (sum of the biomass of C. bilbaoana and S. atrocinerea), but had a great impact on the individual biomass of each plant species. The AMF species with greater extracted ERM lengths (G. mosseae BEG198, G. claroideum BEG210 and the four mixed AMF) preferentially benefited the plant species with a high mycorrhizal dependency (C. bilbaoana), while the AMF species with the smallest ERM length (G. geosporum BEG199) benefited the plant species with a low mycorrhizal dependency (S. atrocinerea). Seed production of C. bilbaoana was only observed in plants inoculated with G. mosseae BEG198, G. claroideum BEG210 or the mixture of the four AMF. Our results show that AMF play an important role in the reproduction of C. bilbaoana coexisting with S. atrocinerea in the alkaline sediment and have the potential to stimulate or completely inhibit seed production. The community composition of native AMF and the length of the mycelium they produce spreading from roots into the surrounding soil can be determinant of the coexistence of naturally co-occurring plant species.

The presence of actinorhizas and arbuscular mycorrhizas may reduce plant stresses caused by adverse soil conditions. A greenhouse experiment was conducted using a sediment with a high pH, resulting from the disposal of waste originated at an acetylene and polyvinylchloride factory, in which Black alder (Alnus glutinosa) seedlings were inoculated either with Glomus intraradices BEG163 (originally isolated from the same sediment), Frankia spp. or both symbionts. After a 6-month growth period, plants inoculated with both symbionts had significantly greater leaf area, shoot height and total biomass when compared with the uninoculated control, the Frankia spp. and the G. intraradices treatments alone. In dual inoculated plants the N and P leaf content was significantly increased. A defoliation experiment was performed to evaluate the stress recovery of A. glutinosa and plants inoculated with both symbionts had a faster leaf regrowth and produced greater numbers of leaves. The dual inoculation resulted in greater numbers of and larger root nodules than when inoculated with Frankia spp. alone. The length and NADH diaphorase activity of the extraradical mycelium of G. intraradices was also significantly greater when dual inoculation was performed. The inoculation with Frankia spp. alone was shown to improve A. glutinosa growth, whereas G. intraradices alone had no positive effect under these environmental conditions. However, when the two symbionts were inoculated together a synergistic effect was observed resulting in a greater benefit for the plants and for both symbionts. The relevance of these findings for the phytorestoration of anthropogenic stressed sediments with high pH is discussed.

Morphological and molecular characters were analysed to investigate diversity within isolates of the Glomus claroideum/Glomus etunicatum species group in the genus Glomus. The inter- and intra-isolate sequence diversity of the large subunit (LSU) rRNA gene D2 region of eight isolates of G. claroideum and G. etunicatum was studied using PCR-single strand conformational polymorphism (SSCP)-sequencing. In addition, two isolates recently obtained from Southern China were included in the analysis to allow for a wider geographic screening. Single spore DNA isolation confirmed the magnitude of gene diversity found in multispore DNA extractions. An apparent overlap of spore morphological characters was found between G. claroideum and G. etunicatum in some isolates. Analysis of the sequence frequencies in all G. etunicatum and G. claroideum isolates (ten) showed that four LSU D2 sequences, representing 32.1% of the clones analysed for multispore extraction (564) were found to be common to both species, and those sequences were the most abundant in four of the ten isolates analysed. The frequency of these sequences ranged between 23.2% and 87.5% of the clones analysed in each isolate. The implications for the use of phenotypic characters to define species in arbuscular mycorrhizal fungi are discussed. The current position of G. claroideum/G.etunicatum in the taxonomy of the Glomeromycota is also discussed.

A field survey of the arbuscular mycorrhizal status of herbaceous plant species was conducted in a highly alkaline anthropogenic sediment resulting from the disposal of waste from an acetylene and polyvinyl chloride factory. Most plant species found at the site were mycorrhizal and the dominant mycotrophic plant species was Conyza bilbaoana. Fungal species richness was assessed by identification of spores extracted from the sediment and from continuously propagated trap pot cultures. All of the six species of arbuscular mycorrhizal fungi (AMF) found were from the genus Glomus. Glomus intraradices and G. mosseae were found in field-collected sediment samples and also occurred most frequently in trap cultures. To test the symbiotic effectiveness of these two fungi, seedlings of C. bilbaoana were inoculated with either native G. intraradices BEG163 or G. mosseae BEG198 and non-native G. intraradices BEG75 or G. mosseae BEG25 isolates in sterile and non-sterile sediment collected from the study site. All four isolates were able to colonise C. bilbaoana. However, AMF native to the target sediments were generally more effective than the non-native fungi in promoting plant establishment and growth under highly alkaline conditions. The non-native G. intraradices was, however, more effective than the non-native G. mosseae. The results of this study suggest the use of adapted AMF as inoculants for phytorestoration of alkaline anthropogenic-stressed sediments.

Summary The existence of multiple rRNA sequences within AMF single spores and the high levels of variability found in these symbionts for highly conserved genes with low copy numbers have provoked intense controversy over the genetic structure of the Glomeromycota. Lately, two hypotheses have been proposed to explain the source of such a variation, mainly either at internuclear (multigenomic) or intranuclear (polyploidy) levels. Evidence reported by numerous authors suggests that the magnitude of sequence variation, and the presence in single spores of sequences from apparently unrelated taxa within the Glomeromycota, makes the use of one, or very few sequences, from only one or two isolates of AMF insufficient for taxonomic purposes in some species groups. A large‐scale, collaborative study on genetic variation in AMF across different genes could help to address fundamental questions on the biology and genetic structure of these important symbionts.

The objective of this study was to investigate the response of plant species used for reclamation of eroded areas in Iceland to inoculation with mycorrhizal fungi. In a greenhouse trial, Leymus arenarius and Deschampsia beringensis were grown in pots with volcanic ash collected from a site near the Mt. Hekla volcano in Iceland and were inoculated with arbuscular mycorrhizal fungi (AMF) isolates Glomus mosseae BEG25 or Glomus intraradices BEG75. In two field experiments conducted on volcanic tephra fields near Mt. Hekla, a native soil inoculum or commercial inocula TerraVital-D and Terra Vital-G Ecto Mix were compared for efficacy on L. arenarius and Betula pubescens. After four months of growth, the presence of AMF in the pot experiment significantly increased the capacity of grass root systems to bind soil particles. In the field, inoculation significantly increased the number of L. arenarius plants, which emerged from seed and their subsequent survival and growth. Seedlings of B. pubescens grew best following inoculation with ectomycorrhizal fungal (ECMF) inoculum and a subsequent application of inorganic NP-fertilizer. The addition of native soil inoculum had almost no effect on growth of either grass or trees. Our results indicate that reclamation of eroded areas in Iceland could benefit from the use of appropriate mycorrhizal fungi, which might improve plant establishment and growth and increase soil aggregation and stability.

•  The role of arbuscular mycorrhizal fungi (AMF) in arsenate resistance in arbuscular mycorrhizal associations is investigated here for two Glomus spp. isolated from the arsenate-resistant grass Holcus lanatus. •  Glomus mosseae and Glomus caledonium were isolated from H. lanatus growing on an arsenic-contaminated mine-spoil soil. The arsenate resistance of spores was compared with nonmine isolates using a germination assay. Short-term arsenate influx into roots and long-term plant accumulation of arsenic by plants were also investigated in uninfected arsenate resistant and nonresistant plants and in plants infected with mine and nonmine AMF. •  Mine AMF isolates were arsenate resistant compared with nonmine isolates. Resistant and nonresistant G. mosseae both suppressed high-affinity arsenate/phosphate transport into the roots of both resistant and nonresistant H. lanatus. Resistant AMF colonization of resistant H. lanatus growing in contaminated mine spoil reduced arsenate uptake by the host. •  We conclude that AMF have evolved arsenate resistance, and conferred enhanced resistance on H. lanatus.

Most published papers by scientists working in the area of mycorrhizal fungi will mention in the first few lines of their introduction the potential importance of these natural fungi for biotechnology in agriculture and of potential applications of their findings. The growing number of new small to medium sized companies (SMEs) around the world (Sylvia 2001) producing inocula of mycorrhizal fungi indicates that many scientists have seen market opportunities for the commercial use of these fungi to increase in the last decade. Many companies have therefore 'spun-out' of the academic and research world into the business world. It is at this point that, in recent years at least, their products have come under increasing scrutiny by fellow scientists and the end-users alike. Many find that the promises made about their product and the results seen by the end-users are often world's apart. This has led to sweeping generalisations, positive and negative, about the efficacy of mycorrhiza products currently available. As natural biological agents and, for arbuscular mycorrhizal fungi at least, non-axenically culturable fungi, there are great problems in presenting the product in the best state for the target markets. Some have taken the approach of single formulations for every market whilst others produce a range of products for their target buyers. Whatever the approach, it is increasingly likely that greater regulation and controls over the production and selling of such inocula will be introduced in the coming years. This is the reality of the business world but scientists and businesses alike need to begin to look at how a series of `best practices' can be adopted by these SMEs to allow the market to develop. At present regulation of these products varies between countries in Europe with some having very tight regulations e.g. France while others are less demanding. Over-regulation will prevent the development of SMEs and could destroy the market for what is potentially one of a few biotechnologies using natural microbes available for plant production.