Late activation of the 9-oxylipin pathway during arbuscular mycorrhiza formation in tomato and its regulation by jasmonate signalling.

The establishment of an arbuscular mycorrhizal (AM) symbiotic interaction is a successful strategy for the promotion of substantial plant growth, development, and fitness. Numerous studies have supported the hypothesis that plant hormones play an important role in the establishment of functional AM symbiosis. Particular attention has been devoted to jasmonic acid (JA) and its derivates, which are believed to play a major role in AM symbiosis. Jasmonates belong to a diverse class of lipid metabolites known as oxylipins that include other biologically active molecules. Recent transcriptional analyses revealed up-regulation of the oxylipin pathway during AM symbiosis in mycorrhizal tomato roots and indicate a key regulatory role for oxylipins during AM symbiosis in tomato, particularly those derived from the action of 9-lipoxygenases (9-LOXs). Continuing with the tomato as a model, the spatial and temporal expression pattern of genes involved in the 9-LOX pathway during the different stages of AM formation in tomato was analysed. The effects of JA signalling pathway changes on AM fungal colonization were assessed and correlated with the modifications in the transcriptional profiles of 9-LOX genes. The up-regulation of the 9-LOX oxylipin pathway in mycorrhizal wild-type roots seems to depend on a particular degree of AM fungal colonization and is restricted to the colonized part of the roots, suggesting that these genes could play a role in controlling fungal spread in roots. In addition, the results suggest that this strategy of the plant to control AM fungi development within the roots is at least partly dependent on JA pathway activation.

Nitrogen availability drives the effect of Glomus intraradices on the growth of strawberry (Fragaria x ananassa Duch.) plants.

BACKGROUND: On the one hand, the critical nitrogen (N) content curve allows the minimal N content necessary for maximum growth rate at any stage of crop development to be predicted. On the other hand, arbuscular mycorrhizal fungi (AMF) transfer N from the soil to the plants and its growth and activity depends on the availability of soil N. Our objective was to investigate how the availability of N in the soil affects growth and the accumulation of N in inoculated strawberry plants. Root colonisation, dry matter accumulation and the critical N% curve were studied during growth of inoculated and non-inoculated strawberry plants grown at several N levels. RESULTS: (1) The increase in the availability of N augmented root colonisation by AMF. (2) The effect of AMF on plant growth depended on N availability and the plant developmental status. (3) The critical %N curves were fitted by the following equations: %N = 2.81× (DM)(-0.21) (r² = 0.81) and %N = 2.89× (DM)(-0.32) (r² = 0.80) for inoculated and non-inoculated plants, respectively (where DM is the weight of leaf dry matter, in g plant⁻¹). CONCLUSION: N availability was a key factor for root colonisation by AMF and for its contribution to plant growth. The patterns of the critical %N curves suggest that AMF modified the photosynthetic N use efficiency.

A comparison of wild-type, old and modern tomato cultivars in the interaction with the arbuscular mycorrhizal fungus Glomus mosseae and the tomato pathogen Fusarium oxysporum f. sp. lycopersici.

The effect of the arbuscular mycorrhizal symbiosis (AM) varies in plant cultivars. In the present study, we tested whether wild-type, old and modern tomato cultivars differ in the parameters of the AM interaction. Moreover, the bioprotective effect of AM against the soilborne tomato pathogen Fusarium oxysporum f. sp. lycopersici (Fol) was tested in the different cultivars. Ten tomato cultivars were inoculated with the arbuscular mycorrhizal fungus (AMF) Glomus mosseae alone or in combination with Fol. At the end of the experiment, AM root colonization, Fusarium infection, and the plant fresh weight was determined. The tomato cultivars differed in their susceptibility to AMF and Fol, but these differences were not cultivar age dependent. In all the cultivars affected by Fol, mycorrhization showed a bioprotective effect. Independent of the cultivar age, tomato cultivars differ in their susceptibility to AMF and Fol and the bioprotective effect of mycorrhization, indicating that the cultivar age does not affect the AM parameters tested in this study.

Altered pattern of arbuscular mycorrhizal formation in tomato ethylene mutants.

Although no specific role has been demonstrated for ethylene during Arbuscular Mycorrhizal (AM) symbiosis, recent results suggest its participation in the regulation of the AM. Analysis of arbuscular mycorrhizal colonization in the abscisic acid (ABA)-deficient tomato sitiens mutant has shown that ABA deficiency induced ethylene production. It has also been suggested that one of the mechanisms used by ABA to determine susceptibility to fungal infection is negative modulation of the ethylene pathway. In this study, we describe the pattern of mycorrhization in mutant plants with altered ethylene biosynthesis and/or perception pathways. Epinastic (epi) plants with increased ethylene response were unaffected in terms of mycorrhizal frequency, although this mutation had a considerable negative impact on the intensity of mycorrhizal root colonization. The negative impact of the mutation in epi plants on the intensity of mycorrhizal root colonization was associated with a transitory increase in the transcript level of the LeETR6 ethylene receptor gene. On the other hand, ripenenig-inhibitor (rin) tomato mutant plants were positively affected in relation to all the mycorrhizal colonization parameters measured, suggesting that, at least in tomato plants, the regulation of AM formation is mediated by the RIN pathway.

Long-distance transport of signals during symbiosis: are nodule formation and mycorrhization autoregulated in a similar way?

Legumes enter nodule symbioses with nitrogen-fixing bacteria (rhizobia), whereas most flowering plants establish symbiotic associations with arbuscular mycorrhizal (AM) fungi. Once first steps of symbiosis are initiated, nodule formation and mycorrhization in legumes is negatively controlled by a shoot-derived inhibitor (SDI), a phenomenon termed autoregulation. According to current views, autoregulation of nodulation and mycorrhization in legumes is regulated in a similar way. CLE peptides induced in response to rhizobial nodulation signals (Nod factors) have been proposed to represent the ascending long-distance signals to the shoot. Although not proven yet, these CLE peptides are likely perceived by leucine-rich repeat (LRR) autoregulation receptor kinases in the shoot. Autoregulation of mycorrhization in non-legumes is reminiscent to the phenomenon of "systemic acquired resistance" in plant-pathogen interactions.

Ethylene-dependent/ethylene-independent ABA regulation of tomato plants colonized by arbuscular mycorrhiza fungi.

We investigated the relationship between ABA and ethylene regulating the formation of the arbuscular mycorrhiza (AM) symbiosis in tomato (Solanum lycopersicum) plants and tried to define the specific roles played by each of these phytohormones in the mycorrhization process. We analysed the impact of ABA biosynthesis inhibition on mycorrhization by Glomus intraradices in transgenic tomato plants with an altered ethylene pathway. We also studied the effects on mycorrhization in sitiens plants treated with the aminoethoxyvinyl glycine hydrochloride (AVG) ethylene biosynthesis inhibitor and supplemented with ABA. In addition, the expression of plant and fungal genes involved in the mycorrhization process was studied. ABA biosynthesis inhibition qualitatively altered the parameters of mycorrhization in accordance with the plant's ethylene perception and ethylene biosynthesis abilities. Inhibition of ABA biosynthesis in wild-type plants negatively affected all the mycorrhization parameters studied, while tomato mutants impaired in ethylene synthesis only showed a reduced arbuscular abundance in mycorrhizal roots. Inhibition of ethylene synthesis in ABA-deficient sitiens plants increased the intensity of mycorrhiza development, while ABA application rescued arbuscule abundance in the root's mycorrhizal zones. The results of our study show an antagonistic interaction between ABA and ethylene, and different roles of each of the two hormones during AM formation. This suggests that a dual ethylene-dependent/ethylene-independent mechanism is involved in ABA regulation of AM formation.

Mycorrhiza-induced trophic cascade enhances fitness and population growth of an acarine predator.

Research on trophic cascades in terrestrial ecosystems has only recently revealed that root-associated organisms interact with organisms living on aboveground plant parts. Arbuscular mycorrhizal (AM) symbiosis is a ubiquitous phenomenon, yet studies on its effect on aboveground natural enemies of herbivores are scarce and mainly deal with plant-mediated rather than herbivore-mediated interactions. Here, we studied herbivore-mediated effects of AM symbiosis on an acarine predator. We measured life history characteristics and population growth rates of Phytoseiulus persimilis preying on two-spotted spider mites, Tetranychus urticae, which were feeding on bean plants colonized or not colonized by the AM fungus Glomus mosseae. All major life history characteristics of P. persimilis, foremost oviposition rate, minimum prey requirements needed to reach adulthood, and developmental time, were positively affected by AM colonization of the host plant of their prey, together resulting in enhanced population growth rates of the predators. Hence, we hypothesize that a bottom-up trophic cascade may counteract the apparent negative effects of mycorrhizal symbiosis when promoting herbivory by promoting the predation of herbivores due to improved prey quality. We argue that this pathway may be involved in stabilizing plant-mycorrhizal symbiosis in ecosystems over time.

The Rhizobium sp. strain NGR234 systemically suppresses arbuscular mycorrhizal root colonization in a split-root system of barley (Hordeum vulgare).

Nitrogen-fixing bacteria (rhizobia) form a nodule symbiosis with legumes, but also induce certain effects on non-host plants. Here, we used a split-root system of barley to examine whether inoculation with Rhizobium sp. strain NGR234 on one side of a split-root system systemically affects arbuscular mycorrhizal (AM) root colonization on the other side. Mutant strains of NGR234 deficient in Nod factor production (strain NGRΔnodABC), perception of flavonoids (strain NGRΔnodD1) and secretion of type 3 effector proteins (strain NGRΩrhcN) were included in this study. Inoculation resulted in a systemic reduction of AM root colonization with all tested strains. However, the suppressive effect of strain NGRΩrhcN was less pronounced. Moreover, levels of salicylic acid, an endogenous molecule related to plant defense, were increased in roots challenged with rhizobia. These data indicate that barley roots perceived NGR234 and that a systemic regulatory mechanism of AM root colonization was activated. The suppressive effect appears to be Nod factor independent, but enhanced by type 3 effector proteins of NGR234.

Root colonisation by the arbuscular mycorrhizal fungus Glomus intraradices alters the quality of strawberry fruits (Fragaria x ananassa Duch.) at different nitrogen levels.

BACKGROUND: Arbuscular mycorrhizal fungi (AMF) increase the uptake of minerals from the soil, thus improving the growth of the host plant. Nitrogen (N) is a main mineral element for plant growth, as it is an essential component of numerous plant compounds affecting fruit quality. The availability of N to plants also affects the AMF-plant interaction, which suggests that the quality of fruits could be affected by both factors. The objective of this study was to evaluate the influence of three N treatments (3, 6 and 18 mmol L(-1)) in combination with inoculation with the AMF Glomus intraradices on the quality of strawberry fruits. The effects of each factor and their interaction were analysed. RESULTS: Nitrogen treatment significantly modified the concentrations of minerals and some phenolic compounds, while mycorrhization significantly affected some colour parameters and the concentrations of most phenolic compounds. Significant differences between fruits of mycorrhizal and non-mycorrhizal plants were found for the majority of phenolic compounds and for some minerals in plants treated with 6 mmol L(-1) N. The respective values of fruits of mycorrhizal plants were higher. CONCLUSION: Nitrogen application modified the effect of mycorrhization on strawberry fruit quality.

Mycorrhization of the notabilis and sitiens tomato mutants in relation to abscisic acid and ethylene contents.

We examined whether the reduced mycorrhization of abscisic acid (ABA)-deficient tomato mutants correlates with their incapacity in ABA biosynthesis and whether this effect is dependent on ethylene production. The mycorrhization of notabilis and sitiens mutants, which have different ABA deficiencies and an excess of ethylene production, was analyzed. Comparative analysis of the ABA-deficient tomato mutants showed both quantitative and qualitative differences in the pattern of arbuscular mycorrhiza (AM) colonization between the two tomato mutant phenotypes. The sitiens mutant showed a great limitation in fungal colonization (mycorrhizal intensity and arbuscule formation) well correlated with their incapacity in ABA biosynthesis. The notabilis plants, which maintained normal ABA levels in roots under our experimental conditions, appeared to be less affected in their capacity for AM formation, and only a decrease in mycorrhizal intensity was noted at the end of the mycorrhization process. Blockage of ABA formation after tungstate application resulted in a reduction in mycorrhization of wild-type tomato plants. The transcript accumulation of the mycorrhiza-responsive LePT4 gene (tomato phosphate transporter) was clearly associated with the ABA content and mycorrhiza development in roots, as the tungstate treatment in wild-type plants and the inherent ABA deficiency in sitiens mutants led to a complete abolishment of their expression. Our results suggest that the decrease in arbuscular abundance in mycorrhizal sitiens roots is directly associated with their ABA biosynthesis deficiency, and the accumulation of ethylene, as a consequence of ABA deficiency in the mutants, primarily affects mycorrhizal intensity.

Root colonization by symbiotic arbuscular mycorrhizal fungi increases sesquiterpenic acid concentrations in Valeriana officinalis L.

In some medicinal plants a specific plant-fungus association, known as arbuscular mycorrhizal (AM) symbiosis, increases the levels of secondary plant metabolites and/or plant growth. In this study, the effects of three different AM treatments on biomass and sesquiterpenic acid concentrations in two IN VITRO propagated genotypes of valerian ( VALERIANA OFFICINALIS L., Valerianaceae) were investigated. Valerenic, acetoxyvalerenic and hydroxyvalerenic acid levels were analyzed in the rhizome and in two root fractions. Two of the AM treatments significantly increased the levels of sesquiterpenic acids in the underground parts of valerian. These treatments, however, influenced the biomass of rhizomes and roots negatively. Therefore this observed increase was not accompanied by an increase in yield of sesquiterpenic acids per plant. Furthermore, one of the two genotypes had remarkably high hydroxyvalerenic acid contents and can be regarded as a hydroxyvalerenic acid chemotype.

Strigolactones, signals for parasitic plants and arbuscular mycorrhizal fungi.

Although strigolactones play a critical role as rhizospheric signaling molecules for the establishment of arbuscular mycorrhizal (AM) symbiosis and for seed germination of parasitic weeds, scarce data are available about interactions between AM fungi and strigolactones. In the present work, we present background data on strigolactones from studies on their seed germination activity on the parasitic weeds Orobanche and Striga, the importance of nitrogen and phosphorus for this seed germination activity, and what this could mean for AM fungi. We also present results on the susceptibility of plants to AM fungi and the possible involvement of strigolactones in this AM susceptibility and discuss the role of strigolactones for the formation and the regulation of the AM symbiosis as well as the possible implication of these compounds as plant signals in other soil-borne plant-microbe interactions.

Arbuscular mycorrhizal symbiosis increases host plant acceptance and population growth rates of the two-spotted spider mite Tetranychus urticae.

Most terrestrial plants live in symbiosis with arbuscular mycorrhizal (AM) fungi. Studies on the direct interaction between plants and mycorrhizal fungi are numerous whereas studies on the indirect interaction between such fungi and herbivores feeding on aboveground plant parts are scarce. We studied the impact of AM symbiosis on host plant choice and life history of an acarine surface piercing-sucking herbivore, the polyphagous two-spotted spider mite Tetranychus urticae. Experiments were performed on detached leaflets taken from common bean plants (Phaseolus vulgaris) colonized or not colonized by the AM fungus Glomus mosseae. T. urticae females were subjected to choice tests between leaves from mycorrhizal and non-mycorrhizal plants. Juvenile survival and development, adult female survival, oviposition rate and offspring sex ratio were measured in order to estimate the population growth parameters of T. urticae on either substrate. Moreover, we analyzed the macro- and micronutrient concentration of the aboveground plant parts. Adult T. urticae females preferentially resided and oviposited on mycorrhizal versus non-mycorrhizal leaflets. AM symbiosis significantly decreased embryonic development time and increased the overall oviposition rate as well as the proportion of female offspring produced during peak oviposition. Altogether, the improved life history parameters resulted in significant changes in net reproductive rate, intrinsic rate of increase, doubling time and finite rate of increase. Aboveground parts of colonized plants showed higher concentrations of P and K whereas Mn and Zn were both found at lower levels. This is the first study documenting the effect of AM symbiosis on the population growth rates of a herbivore, tracking the changes in life history characteristics throughout the life cycle. We discuss the AM-plant-herbivore interaction in relation to plant quality, herbivore feeding type and site and the evolutionary implications in a multi-trophic context.

Effect of mycorrhization on the isoflavone content and the phytoestrogen activity of red clover.

Red clover, known for its estrogenic activity due to its isoflavones content (biochanin A, genistein, daidzein and formononetin), was inoculated with the arbuscular mycorrhizal fungus Glomus mosseae. Once the symbiotic fungus was well established, plants were harvested and we determined the root and shoot dry weight as well as the P-content. In roots and leaves the levels of biochanin A, genistein, daidzein and formononetin were quantified by reversed-phase HPLC and the estrogenic activity of the leaves was measured by a transactivation assay using a yeast two-plasmid system. Mycorrhization increased the levels of biochanin A in the root and the shoot and reduced the levels of genistein in the shoot of red clover. The levels of the other isoflavones were not affected. The shoot biomass of mycorrhizal plants more than doubled compared with non-mycorrhizal control plants, and this growth-stimulating effect of arbuscular mycorrhiza did not affect the estrogenic activity of red clover. In a control P treatment, the biomass of red clover was greatly enhanced. However, the estrogenic activity was reduced. These results suggest that, in contrast to an enhanced shoot biomass production after P application with a reduced estrogenic activity, with arbuscular mycorrhiza the shoot biomass of red clover can be enhanced without a negative effect on estrogenic activity.

Flavonoids and strigolactones in root exudates as signals in symbiotic and pathogenic plant-fungus interactions.

Secondary plant compounds are important signals in several symbiotic and pathogenic plant-microbe interactions. The present review is limited to two groups of secondary plant compounds, flavonoids and strigolactones, which have been reported in root exudates. Data on flavonoids as signaling compounds are available from several symbiotic and pathogenic plant-microbe interactions, whereas only recently initial data on the role of strigolactones as plant signals in the arbuscular mycorrhizal symbiosis have been reported. Data from other plant-microbe interactions and strigolactones are not available yet. In the present article we are focusing on flavonoids in plant-fungal interactions such as the arbuscular mycorrhizal (AM) association and the signaling between different Fusarium species and plants. Moreover the role of strigolactones in the AM association is discussed and new data on the effect of strigolactones on fungi, apart from arbuscular mycorrhizal fungi (AMF), are provided.

The effect of flavones and flavonols on colonization of tomato plants by arbuscular mycorrhizal fungi of the genera Gigaspora and Glomus.

No clear data are available on how flavonoids from different chemical groups affect root colonization by arbuscular mycorrhizal fungi (AMF) and whether flavonoids affecting the presymbiotic growth of AMF also affect root colonization by AMF. In the present work, we compared the effect of flavones (chrysin and luteolin) and flavonols (kaempferol, morin, isorhamnetin, and rutin) on root colonization (number of entry points and degree of root colonization) of tomato plants (Lycopersicum esculentum L.) with the effect of these flavonoids on the presymbiotic growth of these AMF, which has been reported in a recent study. With all tested AMF (Gigaspora rosea, Gigaspora margarita, Glomus mosseae, and Glomus intraradices) a correlation between the number of entry points and the percentage of root colonization was found. When the number of entry points was high, root colonization was also enhanced. Application of the flavones chrysin and luteolin and of the flavonol morin increased the number of entry points and the degree of colonization,whereas the flavonols kaempferol, isorhamnetin, and rutin showed no effect. These results show that in contrast to their effect on the presymbiotic growth of the AMF on the level of root colonization, the tested flavonoids do not exhibit a genus- and species-specificity. Moreover, comparison of our data with the data obtained by J.M. Scervino, M.A. Ponce, R. Erra-Bassells, H. Vierheilig, J.A. Ocampo, and A. Godeas. (2005a. J. Plant Interact. 15: 22-30) indicates that a positive effect on the hyphal growth of AMF does not necessarily result in an enhanced AM root colonization, further indicating that the mode of action of flavonoids at the level of root colonization is more complex.

Abscisic acid determines arbuscule development and functionality in the tomato arbuscular mycorrhiza.

The role of abscisic acid (ABA) during the establishment of the arbuscular mycorrhiza (AM) was studied using ABA sitiens tomato (Lycopersicon esculentum) mutants with reduced ABA concentrations. Sitiens plants and wild-type (WT) plants were colonized by Glomus intraradices. Trypan blue and alkaline phosphatase histochemical staining procedures were used to determine both root colonization and fungal efficiency. Exogenous ABA and silver thiosulfate (STS) were applied to establish the role of ABA and putative antagonistic cross-talk between ABA and ethylene during AM formation, respectively. Sitiens plants were less susceptible to the AM fungus than WT plants. Microscopic observations and arbuscule quantification showed differences in arbuscule morphology between WT and sitiens plants. Both ABA and STS increased susceptibility to the AM fungus in WT and sitiens plants. Fungal alkaline phosphate activity in sitiens mutants was completely restored by ABA application. * The results demonstrate that ABA contributes to the susceptibility of tomato to infection by AM fungi, and that it seems to play an important role in the development of the complete arbuscule and its functionality. Ethylene perception is crucial to AM regulation, and the impairment of mycorrhiza development in ABA-deficient plants is at least partly attributable to ethylene.

Primary colonization and breakdown of igneous rocks by endemic, succulent elephant trees (Pachycormus discolor) of the deserts in Baja California, Mexico.

Trees growing in rocks without soil are uncommon. In two arid regions in Baja California, Mexico, field surveys found large numbers of rock-colonizing elephant trees (Pachycormus discolor (Benth.) Coville ex Standl. (Mexican name: copalquin) growing in igneous rocks (granite and basalt) as primary colonizers without the benefit of soil or with a very small amount of soil generated by their own growth. Many adult trees broke large granite boulders and were capable of wedging, growing in, and colonizing rocks and cliffs made of ancient lava flows. This is the first record of a tree species, apart from the previously recorded cacti, capable of primary colonization of rocks and rock rubble in hot deserts.

Flavonoids exhibit fungal species and genus specific effects on the presymbiotic growth of Gigaspora and Glomus.

The effect of the flavonoids chrysin, isorhamnetin, kaempferol, luteolin, morin and rutin on pre-symbiotic growth, such as spore germination, hyphal length, hyphal branching and the formation of auxiliary cells and secondary spores, of the arbuscular mycorrhizal fungi Gigaspora rosea, G. margarita, Glomus mosseae and G. intraradices was studied. According to the effect on each fungal growth parameter, the tested compounds could be classified to be genus and/or species specific or specific, for a certain developmental stage of pre-symbiotic growth. A clear arbuscular mycorrhizal genus specific, and even species specific, effect of some flavonoids was observed. However, this specificity of a flavonoid could not be generalized but differs depending on the pre-symbiotic stage of the AM fungus. Moreover, our show that for a better understanding of the role of compounds in the AM symbiosis, studies should not be conducted only with one AM fungus looking at one fungal growth parameter such as spore germination or hyphal length, but should be wider, including several growth parameters and several AM fungi.

Arbuscular mycorrhizal colonization of tomato by Gigaspora and Glomus species in the presence of root flavonoids.

The effect of flavonoids isolated from arbuscular mycorrhizal (AM) colonized and noncolonized clover roots on the number of entry points and percentage of root colonization of tomato (Lycopersicum esculentum L.) by Gigaspora rosea, Gi margarita, Glomus mosseae and G. intrarradices symbionts was determined. With fungi of both genera, a correlation between the number of entry points and the percentage of root colonization was found in the presence of some of the tested flavonoids. The flavonoids acacetin and rhamnetin, present in AM clover roots, inhibited the formation of AM penetration structures and the AM colonization of tomato roots, whereas the flavonoid 5,6,7,8,9-hydroxy chalcone, which could not be detected in AM clover root, inhibited both parameters. The flavonoid quercetin, which was present in AM clover roots, stimulated the penetration and root colonization of tomato by Gigaspora. However, the flavonoids 5,6,7,8-hydroxy-4'-methoxy flavone and 3,5,6,7,4'-hydroxy flavone, which was not found in AM clover root, increased the number of entry points and the AM colonization of tomato roots by Gigaspora. These results indicated that flavonoids could be imnplicated in the process of regulation of AM colonization in plant root, but its role is highly complex and depend not only on flavonoids, but also on AM fungal genus or even species.