Cristiana Sbrana

Nonself vegetative fusion and genetic exchange in the arbuscular mycorrhizal fungus Glomus intraradices.

Arbuscular mycorrhizal fungi (AMF) form symbioses with the majority of plants and form extensive underground hyphal networks simultaneously connecting the roots of different plant species. No empirical evidence exists for either anastomosis between genetically different AMF or genetic exchange.Five isolates of one population of Glomus intraradices were used to study anastomosis between hyphae of germinating spores. We show that genetically distinct AMF, from the same field, anastomose, resulting in viable cytoplasmic connections through which genetic exchange could potentially occur.Pairs of genetically different isolates were then co-cultured in an in vitro system.Freshly produced spores were individually germinated to establish new cultures.Using several molecular tools, we show that genetic exchange occurred between genetically different AMF. Specific genetic markers from each parent were transmitted to the progeny. The progeny were viable, forming symbioses with plant roots. The phenotypes of some of the progeny were significantly different from either parent.Our results indicate that considerable promiscuity could occur in these fungi because nine out of 10 combinations of different isolates anastomosed. The ability to perform genetic crosses between AMF experimentally lays a foundation for understanding the genetics and evolutionary biology of these important plants symbionts.

Meeting a non-host: the behaviour of AM fungi

Abstract Arbuscular mycorrhizal (AM) fungi are obligately biotrophic organisms that live symbiotically with the roots of most plants. The establishment of a functional symbiosis between AM fungi and host plants involves a sequence of recognition events leading to the morphological and physiological integration of the two symbionts. The developmental switches in the fungi are triggered by host signals which induce changes in gene expression and a process leading to unequivocal recognition between the two partners of the symbiosis. It has been calculated that about 80% of plant families from all phyla of land plants are hosts of AM fungi. The remaining plant species are either non-mycorrhizal or hosts of mycorrhizas other than the arbuscular type. Non-host plants have been used to obtain information on the factors regulating the development of a functional symbiosis. The aim of this present review is to highlight present-day knowledge of the fungal developmental switches involved in the process of host/non-host discrimination. The following stages of the life cycle of AM fungi are analysed in detail: spore germination, presymbiotic mycelial growth, differential branching pattern and chemotropism, appressorium formation, root colonization.

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.

Development of a model system to assess the impact of genetically modified corn and aubergine plants on arbuscular mycorrhizal fungi

We developed an experimental model system to monitor the impact of generically modified (GM) plants on arbuscular mycorrhizal (AM) fungi, a group of non-target soil microorganisms, fundamental for soil fertility and plant nutrition. The system allowed us to study the effects of root exudates of both commercial Bt corn and aubergine plants expressing Dm-AMP1 defensin on different stages of the life cycle of the AM fungal species G. mosseae. Root exudates of Bt 176 corn significantly reduced pre-symbiotic hyphal growth, compared to Bt 11 and non-transgenic plants. No differences were found in mycelial growth in the presence of Dm-AMP1 and control plant root exudates. Differential hyphal morphogenesis occurred irrespective of the plant line, suggesting that both exuded Bt toxin and defensin do not interfere with fungal host recognition mechanisms. Bt 176 affected the regular development of appressoria, 36% of which failed to produce viable infection pegs. Our experimental model system represents an easy assay for testing the impact of GM plants on non-target soil-borne AM fungi.

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.

The effect of mycorrhizal infection on survival and growth renewal of micropropagated fruit rootstocks

Apple, peach and plum rootstocks were inoculated with the arbuscular mycorrhizal fungus Glomus sp. strain A6 on transplanting from in vitro to in vivo culture. The optimal root length for effective infection, assessed in apple rootstock M 25, was 0.1–1.5 cm, corresponding to the beginning of root elongation. When inoculated at this stage, plants showed maximal growth increase and survival. Mycorrhizal infection of the Mr. S. 2/5 rootstock induced earlier growth renewal after transplanting than in the controls. These results confirm previous reports that mycorrhizal inoculation, performed during transplantation from in vitro to in vivo culture, can enhance both the growth and the survival of plants.

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.

The Impact of Genetically Modified Crops on Soil Microbial Communities

Genetically modified (GM) plants represent a potential benefit for environmentally friendly agriculture and human health. Though, poor knowledge is available on potential hazards posed by unintended modifications occurring during genetic manipulation. The increasing amount of reports on ecological risks and benefits of GM plants stresses the need for experimental works aimed at evaluating the impact of GM crops on natural and agro-ecosystems. Major environmental risks associated with GM crops include their potential impact on non-target soil microorganisms playing a fundamental role in crop residues degradation and in biogeochemical cycles. Recent works assessed the effects of GM crops on soil microbial communities on the basis of case-by-case studies, using multimodal experimental approaches involving different target and non-target organisms. Experimental evidences discussed in this review confirm that a precautionary approach should be adopted, by taking into account the risks associated with the unpredictability of transformation events, of their pleiotropic effects and of the fate of transgenes in natural and agro-ecosystems, weighing benefits against costs.

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.