Tomato responses to Funneliformis mosseae during the early stages of arbuscular mycorrhizal symbiosis.

The concept of symbiosis can be described as a continuum of interactions between organisms ranging from mutualism to parasitism that can also change over time. Arbuscular mycorrhizal fungi (AMF) are among the most important obligate plant symbionts. Once the symbiosis is well established, mycorrhizal plants are more tolerant to biotic or abiotic stresses, so the AMF relationship with the host plant is generally described as mutualistic. However, little is known about AMF effects on the plant during the early stages of root colonization. The aim of this work was to assess the type of interaction (mutualistic or parasitic) between the arbuscular mycorrhizal (AM) fungus Funelliformis mosseae and Solanum lycopersicum cv. Rio Grande plants, at 7, 14, 21, and 28 days after inoculation (DAI), considering that in the adopted experimental design (one plant per pot), the seedling was the only carbon source for fungus development in the absence of common mycorrhizal networks with other plants. At each harvest, mycorrhizal colonization, shoot and root weights, morphometric parameters, and photosynthetic efficiency were evaluated. The presence of the AM fungus in the tomato root system was observed starting from the 14th DAI, and its level increased over time. Few effects of the fungus presence on the considered parameters were observed, and no stress symptoms ever appeared; so, we can state that the fungus behaved as a mutualistic symbiont during the early stages of plant growth. Moreover, a trend towards a positive effect on plant growth was observed at 28 DAI in mycorrhizal plants.

Exploring the survival tactics and plant growth promising traits of root-associated bacterial strains under Cd and Pb stress: A modelling based approach.

The study represents a microbial method for reducing heavy metal stress in terrestrial environment. Two rhizobacterial strains Pantoea agglomerance (PC1) and Pseudomonas aeruginosa (SA) having the ability to tolerate Cd2+ and Pb2+ ions stress, were employed in this study. The growth promotion and survival tactics of the strains under metal stress were explored through kinetic growth model using logistic equation, Luedeking-Piret model and Box Behnken design. Study also involves the interaction of strains with Zea mays L. under Cd2+ and Pb2+ ions stress. Results revealed that both strains have the potential to tolerate 500 mg L-1 of Cd2+ and Pb2+, ions and maintained the plant growth promoting traits. The Luedeking-Piret model estimated the maximum value of IAA on biomass growth (YP/X) 5.377 µg g-1 and 10.3 µg g-1 under Cd2+ ions, while 7.742 µg g-1 and 18.071 µg g-1 under Pb2+ ions stress for strains SA and PC1, respectively. Further, phosphate solubilization activity was optimized with the help of response surface methodology using Box Behnken Design. The optimum solubilization by strain PC1 and SA was achieved at 100 and 150 mg L-1 of Cd2+, and 150 and 200 mg L-1 of Pb2+ ion concentration at the pH range 6.75 and 7.5 respectively. The interactive study with Zea mays L. showed significant increase in seed germination in the presence of Cd2+ and Pb2+ ions thereby proving them as potent plant growth promoters and metal stress reducing biological agents. Hence, the findings of the study suggest that rhizobacterial strains could be a sustainable tool for restoration of metal contaminated sites.

Arbuscular mycorrhizal fungi and plant growth-promoting pseudomonads improve yield, quality and nutritional value of tomato: a field study.

The aim of this work was to assess the effects of plant-beneficial microorganisms (two Pseudomonas strains and a mixed mycorrhizal inoculum, alone or in combination) on the quality of tomato fruits of plants grown in the field and subjected to reduced fertilization. Pseudomonas strain 19Fv1T was newly characterized during this study. The size and quality of the fruits (concentration of sugars, organic acids and vitamin C) were assessed. The microorganisms positively affected the flower and fruit production and the concentrations of sugars and vitamins in the tomato fruits. In particular, the most important effect induced by arbuscular mycorrhizal (AM) fungi was an improvement of citric acid concentration, while bacteria positively modulated sugar production and the sweetness of the tomatoes. The novelty of the present work is the application of soil microorganisms in the field, in a real industrial tomato farm. This approach provided direct information about the application of inocula, allowed the reduction of chemical inputs and positively influenced tomato quality.

AM fungi and PGP pseudomonads increase flowering, fruit production, and vitamin content in strawberry grown at low nitrogen and phosphorus levels.

There is increasing interest in the quality of crops because of the implications concerning health, economic revenue, and food quality. Here we tested if inoculation with a mixture of arbuscular mycorrhizal fungi (AMF) and/or two strains of plant growth-promoting bacteria (PGPB), in conditions of reduced chemical inputs, affects the quality and yield of strawberry fruits. Fruit quality was measured by concentrations of soluble sugars, various organic acids, and two vitamins (ascorbic and folic acid). Co-inoculation with the AMF and each of the two PGPB resulted in increased flower and fruit production, larger fruit size, and higher concentrations of sugars and ascorbic and folic acid in comparison with fruits of uninoculated plants. These results provide further evidence that rhizospheric microorganisms affect fruit crop quality and show that they do so even under conditions of reduced chemical fertilization and can thus be exploited for sustainable agriculture.

Maize development and grain quality are differentially affected by mycorrhizal fungi and a growth-promoting pseudomonad in the field.

Arbuscular mycorrhizal (AM) fungi and plant growth-promoting bacteria (PGPB) can increase the growth and yield of major crops, and improve the quality of fruits and leaves. However, little is known about their impact on seed composition. Plants were inoculated with AM fungi and/or the bacterial strain Pseudomonas fluorescens Pf4 and harvested after 7 months of growth in open-field conditions. Plant growth parameters were measured (biomass, length and circumference of spikes, number of grains per cob, grain yield, and grain size) and protein, lipid, and starch content in grains were determined. Plant growth and yield were increased by inoculation with the microorganisms. Moreover, spikes and grains of inoculated plants were bigger than those produced by uninoculated plants. Regarding grain composition, the bacterial strain increased grain starch content, especially the digestible components, whereas AM fungi-enhanced protein, especially zein, content. Plant inoculation with the fluorescent pseudomonad and mycorrhizal fungi resulted in additive effects on grain composition. Overall, results showed that the bacterial strain and the AM fungi promoted maize growth cultivated in field conditions and differentially affected the grain nutritional content. Consequently, targeted plant inoculation with beneficial microorganisms can lead to commodities fulfilling consumer and industrial requirements.

Development of a new method for detection and identification of Oenococcus oeni bacteriophages based on endolysin gene sequence and randomly amplified polymorphic DNA.

Malolactic fermentation (MLF) is a biochemical transformation conducted by lactic acid bacteria (LAB) that occurs in wine at the end of alcoholic fermentation. Oenococcus oeni is the main species responsible for MLF in most wines. As in other fermented foods, where bacteriophages represent a potential risk for the fermentative process, O. oeni bacteriophages have been reported to be a possible cause of unsuccessful MLF in wine. Thus, preparation of commercial starters that take into account the different sensitivities of O. oeni strains to different phages would be advisable. However, currently, no methods have been described to identify phages infecting O. oeni. In this study, two factors are addressed: detection and typing of bacteriophages. First, a simple PCR method was devised targeting a conserved region of the endolysin (lys) gene to detect temperate O. oeni bacteriophages. For this purpose, 37 O. oeni strains isolated from Italian wines during different phases of the vinification process were analyzed by PCR for the presence of the lys gene, and 25 strains gave a band of the expected size (1,160 bp). This is the first method to be developed that allows identification of lysogenic O. oeni strains without the need for time-consuming phage bacterial-lysis induction methods. Moreover, a phylogenetic analysis was conducted to type bacteriophages. After the treatment of bacteria with UV light, lysis was obtained for 15 strains, and the 15 phage DNAs isolated were subjected to two randomly amplified polymorphic DNA (RAPD)-PCRs. By combining the RAPD profiles and lys sequences, 12 different O. oeni phages were clearly distinguished.

Arbuscular mycorrhizal fungi and plant growth-promoting pseudomonads increases anthocyanin concentration in strawberry fruits (Fragaria x ananassa var. Selva) in conditions of reduced fertilization.

Anthocyanins are a group of common phenolic compounds in plants. They are mainly detected in flowers and fruits, are believed to play different important roles such as in the attraction of animals and seed dispersal, and also in the increase of the antioxidant response in tissues directly or indirectly affected by biotic or abiotic stress factors. As a major group of secondary metabolites in plants commonly consumed as food, they are of importance in both the food industry and human nutrition. It is known that arbuscular mycorrhizal (AM) fungi can influence the plant secondary metabolic pathways such as the synthesis of essential oils in aromatic plants, of secondary metabolites in roots, and increase flavonoid concentration. Plant Growth-Promoting Bacteria (PGPB) are able to increase plant growth, improving plant nutrition and supporting plant development under natural or stressed conditions. Various studies confirmed that a number of bacterial species living on and inside the root system are beneficial for plant growth, yield and crop quality. In this work it is shown that inoculation with AM fungi and/or with selected and tested Pseudomonas strains, under conditions of reduced fertilization, increases anthocyanin concentration in the fruits of strawberry.

Native AMF Communities in an Italian Vineyard at Two Different Phenological Stages of Vitis vinifera.

Arbuscular mycorrhizal fungi (AMF) are beneficial soil microorganisms that can establish symbiotic associations with Vitis vinifera roots, resulting in positive effects on grapevine performance, both in terms of water use efficiency, nutrient uptake, and replant success. Grapevine is an important perennial crop cultivated worldwide, especially in Mediterranean countries. In Italy, Piedmont is one of the regions with the longest winemaking tradition. In the present study, we characterized the AMF communities of the soil associated or not with the roots of V. vinifera cv. Pinot Noir cultivated in a vineyard subjected to conventional management using 454 Roche sequencing technology. Samplings were performed at two plant phenological stages (flowering and early fruit development). The AMF community was dominated by members of the family Glomeraceae, with a prevalence of the genus Glomus and the species Rhizophagus intraradices and Rhizophagus irregularis. On the contrary, the genus Archaeospora was the only one belonging to the family Archaeosporaceae. Since different AMF communities occur in the two considered soils, independently from the plant phenological stage, a probable role of V. vinifera in determining the AMF populations associated to its roots has been highlighted.

Proteomic analysis of Pteris vittata fronds: two arbuscular mycorrhizal fungi differentially modulate protein expression under arsenic contamination.

Arbuscular mycorrhizae (AM) are the most widespread mutualistic symbioses between the roots of most land plants and a phylum of soil fungi. AM are known to influence plant performance by improving mineral nutrition, protecting against pathogens and enhancing resistance or tolerance to biotic and abiotic stresses. The aim of this study was to investigate the frond proteome of the arsenic hyperaccumulator fern Pteris vittata in plants that had been inoculated with one of the two AM fungi (Glomus mosseae or Gigaspora margarita) with and without arsenic treatment. A protective role for AM fungi colonisation in the absence of arsenic was indicated by the down-regulation of oxidative damage-related proteins. Arsenic treatment of mycorrhizal ferns induced the differential expression of 130 leaf proteins with specific responses in G. mosseae- and Gi. margarita-colonised plants. Up-regulation of multiple forms of glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and enolase, primarily in G. mosseae-inoculated plants, suggests a central role for glycolytic enzymes in arsenic metabolism. Moreover, a putative arsenic transporter, PgPOR29, has been identified as an up-regulated protein by arsenic treatment.

Effects of Pseudomonas putida S1Pf1Rif against chrysanthemum yellows phytoplasma infection.

Phytoplasmas cause damage on a number of plant species leading to relevant economical loss. Up to now, strategies to limit their spread led to only partial success. In this context, the use of plant-beneficial bacteria to control phytoplasmas has never been explored. The aim of this work was to assess the effect of Pseudomonas putida S1Pf1Rif against chrysanthemum yellows phytoplasma (CYP) infection of daisy. Plant biomass, root architecture, symptom severity, phytoplasma titer, and viability were evaluated in inoculated and control plants. CYP reduced plant growth and root development. Although the phytoplasma titer in young apical leaves was not affected by inoculation with S1Pf1Rif, the pseudomonad improved plant growth of CYP-infected plants. Whereas CYP titer increased over time in uninoculated plants, its viability decreased, regardless of the presence of P. putida S1Pf1Rif. Finally, phytoplasma cells in fully developed leaves of CYP-infected plants inoculated with S1Pf1Rif often appeared degenerated. Overall, our results indicate that P. putida S1Pf1Rif is able to alleviate the disease, although it does not affect the presence of viable phytoplasmas in young, developing leaves of the infected plants.

Screening for heavy metal accumulators amongst autochtonous plants in a polluted site in Italy.

The site around ACNA factory (Northern Italy) is characterized by multi-metal contamination, therefore it can be considered as a source of autochthonous plants able to tolerate or accumulate heavy metals (HMs). The hill A5, a waste dump of the chemical factory, was chosen as the study area, in order to assess the metal accumulation ability of the vegetation growing under HM stress. The plant species, biodiversity and health were related to the concentration of HMs in four areas of the hill A5, and to the metal accumulation in shoots and roots. Uptake of HMs occurred at different extent in the various plant species and differed according to the considered organ and metal. Polygonum aviculare hyperaccumulated Hg in the shoot suggesting its possible exploitation in phytoextraction. A number of species, that can be useful in phytoremediation plans, accumulated simultaneously more than two heavy metals both in the shoot and in the root.

Zinc tolerance and accumulation in the ferns Polypodium cambricum L. and Pteris vittata L.

Zn uptake and compartmentalisation were studied in two ferns, the European Polypodium cambricum L., a possible Zn tolerant, and the sub-tropical Pteris vittata L., an As accumulator also able to accumulate Zn. Ferns growing in hydroponic systems were exposed to Zn concentrations ranging from non-toxic to lethal doses (0, 50, 125, 250, 500 mg kg(-1) as ZnSO4). After treatments, the following analyses were made: photosynthetic efficiency (Handy PEA), anatomical symptoms (optical and scanning electron microscopy), determination of Zn in fronds, rhizome and roots (atomic emission spectrometry, ICP-AES). Both species showed high bioconcentration and bioaccumulation factors, but low translocation factor, indicating Zn sequestration in the root/rhizome system. P. cambricum was more resistant to Zn, while P. vittata suffered from unrestricted uptake leading to macro- and microscopical damages and plant death. Data suggest that P. cambricum could be suitable for phytostabilisation of Zn-contaminated soils in temperate areas.

Phytoremediation of a Highly Arsenic Polluted Site, Using Pteris vittata L. and Arbuscular Mycorrhizal Fungi.

Phytoremediation is a promising green technique for the restoration of a polluted environment, but there is often a gap between lab and field experiments. The fern, Pteris vittata L., can tolerate a high soil arsenic concentration and rapidly accumulate the metalloid in its fronds. Arbuscular mycorrhizal fungi (AMF) are mutualistic fungi that form a symbiosis with most land plants' roots, improve their growth, and induce stress tolerance. This paper reports the results obtained using P. vittata inoculated with AMF, to extract Arsenic (As) from an industrial site highly contaminated also by other pollutants. Two experiments have been performed. In the first one, AMF colonized ferns were grown for two years under controlled conditions in soil coming from the metallurgic site. Positive effects on plant health and As phytoextraction and accumulation were detected. Then, considering these results, we performed a three year in situ experiment in the industrial site, to assess the remediation of As at two different depths. Our results show that the colonization of P. vittata with AMF improved the remediation process of As with a significant impact on the depth 0-0.2 m.

Discovering the bacteriome of Vitis vinifera cv. Pinot Noir in a conventionally managed vineyard.

The structure of the bacteriome associated with grapevine roots can affect plant development, health and grape quality. We previously investigated the bacterial biodiversity of the Vitis vinifera cv. Pinot Noir rhizosphere in a vineyard subjected to integrated pest management. The aim of this work is to characterize the bacteriome of V. vinifera cv. Pinot Noir in a conventionally managed vineyard using a metabarcoding approach. Comparisons between the microbial community structure in bulk soil and rhizosphere (variable space) were performed and shifts of bacteriome according to two sampling times (variable time) were characterized. Bacterial biodiversity was higher at the second than at the first sampling and did not differ according to the variable space. Actinobacteria was the dominant class, with Gaiella as the most represented genus in all the samples. Among Proteobacteria, the most represented classes were Alpha, Beta and Gamma-Proteobacteria, with higher abundance at the second than at the first sampling time. Bradyrhizobium was the most frequent genus among Alpha-Proteobacteria, while Burkholderia was the predominant Beta-Proteobacteria. Among Firmicutes, the frequency of Staphylococcus was higher than 60% in bulk soil and rhizosphere. Finally, the sampling time can be considered as one of the drivers responsible for the bacteriome variations assessed.

Proteomic analysis of Arabidopsis halleri shoots in response to the heavy metals cadmium and zinc and rhizosphere microorganisms.

Arabidopsis halleri has the rare ability to colonize heavy metal-polluted sites and is an emerging model for research on adaptation and metal hyperaccumulation. The aim of this study was to analyze the effect of plant-microbe interaction on the accumulation of cadmium (Cd) and zinc (Zn) in shoots of an ecotype of A. halleri grown in heavy metal-contaminated soil and to compare the shoot proteome of plants grown solely in the presence of Cd and Zn or in the presence of these two metals and the autochthonous soil rhizosphere-derived microorganisms. The results of this analysis emphasized the role of plant-microbe interaction in shoot metal accumulation. Differences in protein expression pattern, identified by a proteomic approach involving 2-DE and MS, indicated a general upregulation of photosynthesis-related proteins in plants exposed to metals and to metals plus microorganisms, suggesting that metal accumulation in shoots is an energy-demanding process. The analysis also showed that proteins involved in plant defense mechanisms were downregulated indicating that heavy metals accumulation in leaves supplies a protection system and highlights a cross-talk between heavy metal signaling and defense signaling.

Expression of the PsMTA1 gene in white poplar engineered with the MAT system is associated with heavy metal tolerance and protection against 8-hydroxy-2'-deoxyguanosine mediated-DNA damage.

Marker-free transgenic white poplar (Populus alba L., cv 'Villafranca') plants, expressing the PsMT (A1) gene from Pisum sativum for a metallothionein-like protein, were produced by Agrobacterium tumefaciens-mediated transformation. The 35SCaMV-PsMT (A1)-NosT cassette was inserted into the ipt-type vector pMAT22. The occurrence of the abnormal ipt-shooty phenotype allowed the visual selection of transformants, while the yeast site-specific recombination R/RS system was responsible for the excision of the undesired vector sequences with the consequent recovery of normal marker-free transgenic plants. Molecular analyses confirmed the presence of the 35SCaMV-PsMT (A1)-NosT cassette and transgene expression. Five selected lines were further characterized, revealing the ability to withstand heavy metal toxicity. They survived 0.1 mM CuCl(2), a concentration which strongly affected the nontransgenic plants. Moreover, root development was only slightly affected by the ectopic expression of the transgene. Reactive oxygen species were accumulated to a lower extent in leaf tissues of multi-auto-transformation (MAT)-PsMT(A1) plants exposed to copper and zinc, compared to control plants. Tolerance to photo-oxidative stress induced by paraquat was another distinctive feature of the MAT-PsMT(A1) lines. Finally, low levels of DNA damage were detected by quantifying the amounts of 8-hydroxy-2'-deoxyguanosine in leaf tissues of the transgenic plants exposed to copper.

Beneficial role of plant growth promoting bacteria and arbuscular mycorrhizal fungi on plant responses to heavy metal stress.

Heavy metal pollution is a major worldwide environmental concern that has recently motivated researchers to develop a variety of novel approaches towards its cleanup. As an alternative to traditional physical and chemical methods of environmental cleanup, scientists have developed phytoremediation approaches that include the use of plants to remove or render harmless a range of compounds. Both plant growth promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF) can be used to facilitate the process of phytoremediation and the growth of plants in metal-contaminated soils. This review focuses on the recent literature dealing with the effects of plant growth-promoting bacteria and AM fungi on the response of plants to heavy metal stress and points the way to strategies that may facilitate the practical realization of this technology.

A review on global metal accumulators-mechanism, enhancement, commercial application, and research trend.

The biosphere is polluted with metals due to burning of fossil fuels, pesticides, fertilizers, and mining. The metals interfere with soil conservations such as contaminating aqueous waste streams and groundwater, and the evidence of this has been recorded since 1900. Heavy metals also impact human health; therefore, the emancipation of the environment from these environmental pollutants is critical. Traditionally, techniques to remove these metals include soil washing, removal, and excavation. Metal-accumulating plants could be utilized to remove these metal pollutants which would be an alternative option that would simultaneously benefit commercially and at the same time clean the environment from these pollutants. Commercial application of pollutant metals includes biofortification, phytomining, phytoremediation, and intercropping. This review discusses about the metal-accumulating plants, mechanism of metal accumulation, enhancement of metal accumulation, potential commercial applications, research trends, and research progress to enhance the metal accumulation, benefits, and limitations of metal accumulators. The review identified that the metal accumulator plants only survive in low or medium polluted environments with heavy metals. Also, more research is required about metal accumulators in terms of genetics, breeding potential, agronomics, and the disease spectrum. Moreover, metal accumulators' ability to uptake metals need to be optimized by enhancing metal transportation, transformation, tolerance to toxicity, and volatilization in the plant. This review would benefit the industries and environment management authorities as it provides up-to-date research information about the metal accumulators, limitation of the technology, and what could be done to improve the metal enhancement in the future.

Preferential colonization of Solanum tuberosum L. roots by the fungus Glomus intraradices in arable soil of a potato farming area.

The symbiosis between plant roots and arbuscular mycorrhizal (AM) fungi has been shown to affect both the diversity and productivity of agricultural communities. In this study, we characterized the AM fungal communities of Solanum tuberosum L. (potato) roots and of the bulk soil in two nearby areas of northern Italy, in order to verify if land use practices had selected any particular AM fungus with specificity to potato plants. The AM fungal large-subunit (LSU) rRNA genes were subjected to nested PCR, cloning, sequencing, and phylogenetic analyses. One hundred eighty-three LSU rRNA sequences were analyzed, and eight monophyletic ribotypes, belonging to Glomus groups A and B, were identified. AM fungal communities differed between bulk soil and potato roots, as one AM fungal ribotype, corresponding to Glomus intraradices, was much more frequent in potato roots than in soils (accounting for more than 90% of sequences from potato samples and less than 10% of sequences from soil samples). A semiquantitative heminested PCR with specific primers was used to confirm and quantify the AM fungal abundance observed by cloning. Overall results concerning the biodiversity of AM fungal communities in roots and in bulk soils from the two studied areas suggested that potato roots were preferentially colonized by one AM fungal species, G. intraradices.

Colonization of adventitious roots of Medicago truncatula by Pseudomonas fluorescens C7R12 as affected by arbuscular mycorrhiza.

Pseudomonas fluorescens C7R12 was previously shown to promote colonization of Medicago truncatula roots by Glomus mosseae BEG12. To gain more insight into the interaction between C7R12 and BEG12, the cell organization of C7R12 was characterized on adventitious roots mycorrhized or not with BEG12 and on extraradical hyphae. Bacterial cell observations were made using the immuno-fluorescence technique and confocal laser scanning microscopy. Five types of cell organization, so-called organization types (OT), were identified: small or large single cells, cells by pair and cells in microcolonies or in strings. The frequencies of each OT on the roots were expressed as the percentage of observations in which these OTs were represented. The OT frequencies on mycorrhizal and nonmycorrhizal roots differed significantly. Bacterial cells were more frequently single on mycorrhizal than on nonmycorrhizal roots, and in microcolonies and strings on nonmycorrhizal roots. Furthermore, the root area covered by bacterial cells, as assessed by image analysis, appeared to be significantly lower on mycorrhizal than on nonmycorrhizal roots. C7R12 cells were abundant on extraradical hyphae and organized both as single cells and microcolonies. Taken together, these results suggest that P. fluorescens C7R12 cells were less active and less abundant on mycorrhizal than on nonmycorrhizal roots.