Luciano Avio

Genetic diversity of isolates of Glomus mosseae from different geographic areas detected by vegetative compatibility testing and biochemical and molecular analysis.

ABSTRACT We detected, for the first time, the occurrence of vegetative incompatibility between different isolates of the arbuscular mycorrhizal fungal species Glomus mosseae. Vegetative compatibility tests performed on germlings belonging to the same isolate showed that six geographically different isolates were capable of self-anastomosing, and that the percentage of hyphal contacts leading to fusions ranged from 60 to 85%. Successful anastomoses were characterized by complete fusion of hyphal walls, protoplasm continuity and occurrence of nuclei in the middle of hyphal bridges. No anastomoses could be detected between hyphae belonging to different isolates, which intersected without any reaction in 49 to 68% of contacts. Microscopic examinations detected hyphal incompatibility responses in diverse pairings, consisting of protoplasm retraction from the tips and septum formation in the approaching hyphae, even before physical contact with neighboring hyphae. Interestingly, many hyphal tips showed precontact tropism, suggesting that specific recognition signals may be involved during this stage. The intraspecific genetic diversity of G. mosseae revealed by vegetative compatibility tests was confirmed by total protein profiles and internal transcribed spacer-restriction fragment length polymorphism profiles, which evidenced a higher level of molecular diversity between the two European isolates IMA1 and BEG25 than between IMA1 and the two American isolates. Since arbuscular mycorrhizal fungi lack a tractable genetic system, vegetative compatibility tests may represent an easy assay for the detection of genetically different mycelia and an additional powerful tool for investigating the population structure and genetics of these obligate symbionts.

At the Root of the Wood Wide Web: Self Recognition and Nonself Incompatibility in Mycorrhizal Networks

Arbuscular mycorrhizal (AM) fungi are mutualistic symbionts living in the roots of 80% of land plant species, and developing extensive, belowground extraradical hyphae fundamental for the uptake of soil nutrients and their transfer to host plants. Since AM fungi have a wide host range, they are able to colonize and interconnect contiguous plants by means of hyphae extending from one root system to another. Such hyphae may fuse due to the widespread occurrence of anastomoses, whose formation depends on a highly regulated mechanism of self recognition. Here, we examine evidences of self recognition and nonself incompatibility in hyphal networks formed by AM fungi and discuss recent results showing that the root systems of plants belonging to different species, genera and families may be connected by means of anastomosis formation between extraradical mycorrhizal networks, which can create indefinitely large numbers of belowground fungal linkages within plant communities.

Biotechnology of arbuscular mycorrhizas

Publisher Summary Mycorrhizas are symbiotic associations established between thousands of species of soil-borne fungi and the roots of most terrestrial plant species. This chapter provides an overview and analyzes important data on the main parameters affecting fungal infectivity and efficiency and on fungal ability to survive, multiply and spread in different environments, which may contribute to the biotechnological exploitation and utilization of arbuscular mycorrhizas (AM) fungi in sustainable agriculture and biodiversity conservation. AM fungi represent fundamental factors of plant productivity, because they are capable, by means of extraradical hyphae, to mediate transfer of nutrients from the soil to host plants and between different plants linked by a common mycorrhizal network. Their management appears essential for their use as biofertilizers, for reducing the inputs of chemical fertilizers and pesticides, in the perspectives of sustainable agriculture and of conservation of natural resources. Because, large variations in symbiotic performance have been found in the different host/fungus/soil combinations, a more systematic approach is necessary to detect and select infective and efficient strains to be used for inoculation in diverse host plants and soil conditions.

Beneficial mycorrhizal symbionts affecting the production of health-promoting phytochemicals.

Fresh fruits and vegetables are largely investigated for their content in vitamins, mineral nutrients, dietary fibers, and plant secondary metabolites, collectively called phytochemicals, which play a beneficial role in human health. Quantity and quality of phytochemicals may be detected by using different analytical techniques, providing accurate quantification and identification of single molecules, along with their molecular structures, and allowing metabolome analyses of plant‐based foods. Phytochemicals concentration and profiles are affected by biotic and abiotic factors linked to plant genotype, crop management, harvest season, soil quality, available nutrients, light, and water. Soil health and biological fertility play a key role in the production of safe plant foods, as a result of the action of beneficial soil microorganisms, in particular of the root symbionts arbuscular mycorrhizal fungi. They improve plant nutrition and health and induce changes in secondary metabolism leading to enhanced biosynthesis of health‐promoting phytochemicals, such as polyphenols, carotenoids, flavonoids, phytoestrogens, and to a higher activity of antioxidant enzymes. In this review we discuss reports on health‐promoting phytochemicals and analytical methods used for their identification and quantification in plants, and on arbuscular mycorrhizal fungi impact on fruits and vegetables nutritional and nutraceutical value.

Contrasting effects of cover crops on 'hot spot' arbuscular mycorrhizal fungal communities in organic tomato

Arbuscular mycorrhizal fungal (AMF) communities are fundamental in organic cropping systems where they provide essential agro-ecosystem services, improving soil fertility and sustaining crop production. They are affected by agronomic practices, but still, scanty information is available about the role of specific crops, crop rotations and the use of winter cover crops on the AMF community compositions at the field sites. A field experiment was conducted to elucidate the role of diversified cover crops and AMF inoculation on AMF diversity in organic tomato. Tomato, pre-inoculated at nursery with two AMF isolates, was grown following four cover crop treatments: Indian mustard, hairy vetch, a mixture of seven species and a fallow. Tomato root colonization at flowering was more affected by AMF pre-transplant inoculation than by the cover crop treatments. An enormous species richness was found by morphological spore identification: 58 AMF species belonging to 14 genera, with 46 and 53 species retrieved at the end of cover crop cycle and at tomato harvest, respectively. At both sampling times, AMF spore abundance was highest in hairy vetch, but after tomato harvest, AMF species richness and diversity were lower in hairy vetch than in the cover crop mixture and in the mustard treatments. A higher AMF diversity was found at tomato harvest, compared with the end of the cover crop cycle, independent of the cover crop and pre-transplant AMF inoculation. Our findings suggest that seasonal and environmental factors play a major role on AMF abundance and diversity than short-term agronomic practices, including AMF inoculation. The huge AMF diversity is explained by the field history and the Mediterranean environment, where species characteristic of temperate and sub-tropical climates co-occur.

Mycorrhizal activity and diversity in a long-term organic Mediterranean agroecosystem

In organic agriculture, soil fertility and productivity rely on biological processes carried out by soil microbes, which represent the key elements of agroecosystem functioning. Arbuscular mycorrhizal fungi (AMF), fundamental microorganisms for soil fertility, plant nutrition and health, may play an important role in organic agriculture by compensating for the reduced use of fertilizers and pesticides. Though, AMF activity and diversity following conversion from conventional to organic farming are poorly investigated. Here we studied AMF abundance, diversity and activity in short- and long-term organically and conventionally managed Mediterranean arable agroecosystems. Our results show that both AMF population activity, as assessed by the mycorrhizal inoculum potential (MIP) assay, the percentage of colonized root length of the field crop (maize) and glomalin-related soil protein (GRSP) content were higher in organically managed fields and increased with time since transition to organic farming. Here, we showed an increase of GRSP content in arable organic systems and a strong correlation with soil MIP values. The analysis of AMF spores showed differences among communities of the three microagroecosystems in terms of species richness and composition as suggested by a multivariate analysis. All our data indicate that AMF respond positively to the transition to organic farming by a progressive enhancement of their activity that seems independent from the species richness of the AMF communities. Our study contributes to the understanding of the effects of agricultural managements on AMF, which represent a promising tool for the implementation of sustainable agriculture.

The protein pattern of spores of arbuscular mycorrhizal fungi: comparison of species, isolates and physiological stages

Spore proteins of different species and isolates of arbuscular mycorrhizal (AM) fungi were compared by PAGE. Reproducibility of protein patterns was assessed by using cultures of the same species either grown on different host plants, or produced during successive propagation cycles and stored up to 5 years. The results consistently showed that host species, different generations and storage, did not affect protein profiles, thus validating the accuracy of the method. Comparison among different geographical isolates of the same species revealed consistent protein patterns. The stability and diversity of spore protein profiles suggested that PAGE could be used to discriminate and identify AM fungal species and isolates. By contrast, the physiological state of spores affected the quality and quantity of bands, with germinating spores showing marked profile changes, as compared to quiescent spores, both in denaturating and native analytical conditions. The disappearance of some polypeptides in germinated spores might be related to the occurrence of storage proteins in AM fungi.

A new fungal species forming arbuscular mycorrhizas: Glomus viscosum

A species of Glomus forming arbuscular mycorrhizas was established in pot culture with several plants. The fungus forms abundant hyaline to white spores in loose clusters. In addition to the normal spore wall which is continuous with the wall of the subtending hypha, the spores of this species produce an apparently mucilaginous outer coating, resulting in the adherence of minute soil particles which were shown to be composed of soil particles by EDAX analysis. The fungus is named Glomus viscosum in reference to the sticky outer coating. The spores, germination characteristics, and mycorrhizas are described and illustrated, and the symbiotic behaviour is discussed.

Vesicular-arbuscular mycorrhizal infection of lucerne roots in a cellulose-amended soil

Lucerne plants inoculated with vesicular-arbuscular mycorrhizal fungi were grown in pots in a sandy soil amended or not amended with cellulose. Whatever the endophyte or cellulose sources used, the rate of VAM colonization was lower in amended soil. The inhibition of VAM infection increased with the cellulose concentration in the soil. Sequential harvest experiments showed a clear reduction of root colonization from the early stages of plant growth. The effectiveness of cellulose in reducing VAM colonization was influenced by the addition of N fertilizer.

The occurrence of calcofluor and lectin binding polysaccharides in the outer wall of arbuscular mycorrhizal fungal spores

The hyaline outer wall of some spores of arbuscular mycorrhizal fungi is of important diagnostic value in identifying species but little is known about the chemical characteristics of this wall. With the use of the fluorochrome calcofluor and of the fluorescein-conjugated lectin wheat germ agglutinin we observed that chitin is one of the main components of the hyaline outer wall of four species of Glomus, and that neither chemical nor enzymic treatments of the spores were capable of changing the binding properties of this polysaccharide.