Uptake of zinc and phosphorus by plants is affected by zinc fertiliser material and arbuscular mycorrhizas

Background and Aims

Water solubility of zinc (Zn) fertilisers affects their plant availability. Further, simultaneous application of Zn and phosphorus (P) fertiliser can have antagonistic effects on plant Zn uptake. Arbuscular mycorrhizas (AM) can improve plant Zn and P uptake. We conducted a glasshouse experiment to test the effect of different Zn fertiliser materials, in conjunction with P fertiliser application, and colonisation by AM, on plant nutrition and biomass.

Methods

We grew a mycorrhiza-defective tomato genotype (rmc) and its mycorrhizal wild-type progenitor (76R) in soil with six different Zn fertilisers ranging in water solubility (Zn sulphate, Zn oxide, Zn oxide (nano), Zn phosphate, Zn carbonate, Zn phosphate carbonate), and supplemental P. We measured plant biomass, Zn and P contents, mycorrhizal colonisation and water use efficiency.

Results

Whereas water solubility of the Zn fertilisers was not correlated with plant biomass or Zn uptake, plant Zn and P contents differed among Zn fertiliser treatments. Plant Zn and P uptake was enhanced when supplied as Zn phosphate carbonate. Mycorrhizal plants took up more P than non-mycorrhizal plants; the reverse was true for Zn.

Conclusions

Zinc fertiliser composition and AM have a profound effect on plant Zn and P uptake.     

Field inoculation with arbuscular-mycorrhizal fungi overcomes phosphorus and zinc deficiencies of linseed (Linum usitatissimum) in a vertisol subject to long-fallow disorder

Background and aims

Long-fallow disorder is expressed as exacerbated deficiencies of phosphorus (P) and/or zinc (Zn) in field crops growing after long periods of weed-free fallow. The hypothesis that arbuscular-mycorrhizal fungi (AMF) improve the P and Zn nutrition, and thereby biomass production and seed yield of linseed (Linum usitatissimum) was tested in a field experiment.

Methods

A factorial combination of treatments consisting of ± fumigation, ±AMF inoculation with Glomus spp., ±P and ±Zn fertilisers was used on a long-fallowed vertisol. The use of such methods allowed an absolute comparison of plants growing with and without AMF in the field for the first time in a soil disposed to long-fallow disorder.

Results

Plant biomass, height, P and Zn concentrations and contents, boll number and final seed yield were (a) least in fumigated soil with negligible AMF colonisation of the roots, (b) low initially in long-fallow soil but increased with time as AMF colonisation of the roots developed, and (c) greatest in soil inoculated with AMF cultures. The results showed for the first time in the field that inflows of both P and Zn into linseed roots were highly dependent on %AMF-colonisation (R²?=?0.95 for P and 0.85 for Zn, P?<?0.001) in a soil disposed to long-fallow disorder. Relative field mycorrhizal dependencies without and with P+Zn fertiliser were 85 % and 86 % for biomass and 68 % and 52 % for seed yield respectively.

Conclusions

This research showed in the field that AMF greatly improved the P and Zn nutrition, biomass production and seed yield of linseed growing in a soil disposed to long-fallow disorder. The level of mycorrhizal colonisation of plants suffering from long-fallow disorder can increase during the growing season resulting in improved plant growth and residual AMF inoculum in the soil, and thus it is important for growers to recognise the cause and not terminate a poor crop prematurely in order to sow another. Other positive management options to reduce long fallows and foster AMF include adoption of conservation tillage and opportunity cropping.     

Arbuscular mycorrhizas and their role in plant growth, nitrogen interception and soil gas efflux in an organic production system

Background and aims

Roots and mycorrhizas play an important role in not only plant nutrient acquisition, but also ecosystem nutrient cycling.

Methods

A field experiment was undertaken in which the role of arbuscular mycorrhizas (AM) in the growth and nutrient acquisition of tomato plants was investigated. A mycorrhiza defective mutant of tomato (Solanum lycopersicum L.) (named rmc) and its mycorrhizal wild type progenitor (named 76R) were used to control for the formation of AM. The role of roots and AM in soil N cycling was studied by injecting a ¹⁵N-labelled nitrate solution into surface soil at different distances from the 76R and rmc genotypes of tomato, or in plant free soil. The impacts of mycorrhizal and non-mycorrhizal root systems on soil greenhouse gas (CO₂ and ¹⁴⁺¹⁵N₂O and ¹⁵N₂O) emissions, relative to root free soils, were also studied.

Results

The formation of AM significantly enhanced plant growth and nutrient acquisition, including interception of recently applied NO ₃ ^? . Whereas roots caused a small but significant decrease in ¹⁵N₂O emissions from soils at 23?h after labeling, compared to the root-free treatment, arbuscular mycorrhizal fungi (AMF) had little effect on N₂O emissions. In contrast soil CO₂ emissions were higher in plots containing mycorrhizal root systems, where root biomass was also greater.

Conclusions

Taken together, these data indicate that roots and AMF have an important role to play in plant nutrient acquisition and ecosystem N cycling.     

Interactions between root hair length and arbuscular mycorrhizal colonisation in phosphorus deficient barley (Hordeum vulgare)

Aims

Phosphorus (P) limits crop yield and P-fertilisers are frequently applied to agricultural soils. However, supplies of quality rock phosphate are diminishing. Plants have evolved mechanisms to improve P-acquisition and understanding these could improve the long-term sustainability of agriculture. Here we examined interactions between root hairs and arbuscular mycorrhizal (AM) colonisation in barley (Hordeum vulgare L.).

Methods

Barley mutants exhibiting different root hair phenotypes, wild type barley and narrowleaf plantain (Plantago lanceolata L.) were grown in the glasshouse in P-sufficient and P-deficient treatments and allowed to develop AM colonization from the natural soil community. Plants were harvested after 6 weeks growth and root hair length, AM-fungal colonisation, shoot biomass and P-accumulation measured.

Results

Under P-deficient conditions, root hair length and AM colonisation were negatively related suggesting that resources are allocated to root hairs rather than to AM fungi in response to P-deficiency. There was evidence that barley and narrowleaf plantain employed different strategies to increase P-acquisition under identical conditions, but root hairs were more effective.

Conclusions

This research suggests future barley breeding programmes should focus on maintaining or improving root hair phenotypes and that pursuing enhancements to AM associations under the prevalent agricultural conditions tested here would be ineffectual.     

Ninety-year-, but not single, application of phosphorus fertilizer has a major impact on arbuscular mycorrhizal fungal communities

Background and aims

Arbuscular mycorrhizal (AM) fungi play a significant role in P nutrition of crops in agriculture, but P accumulation in the soil, e.g., application of P-fertilizer, generally reduces AM fungal colonization. The impact of long-term application of chemical fertilizer on AM fungal communities was investigated with respect to the time scale.

Methods

Soils were collected from four plots with different fertilizer management in the long-term experimental field established in 1914. Lotus japonicus was grown in the soils in a greenhouse, while Glycine max was grown in the plots in the field. DNA was extracted from their roots, and the diversity and community compositions were analyzed based on occurrence of the AM fungal phylotypes defined by sequence similarity in the LSU rDNA.

Results

The 90-year-application of N and K in the absence of P increased AM fungal diversity and resulted in formation of a distinctive fungal community compared with those in the other treatments. This effect was not cancelled by single application of P. Whereas the impact of balanced application of N, P, and K was ambiguous.

Conclusion

These observations suggest that the presence/absence of P-fertilizer has a major impact on AM fungal communities, but the action may appear only on a long time scale.     

Interactive influence of light intensity and soil fertility on root-associated arbuscular mycorrhizal fungi

Background and aims

Soil nutrients and light have major effects on the economics of arbuscular mycorrhizal (AM) symbioses. This study tests the main and interactive effects of soil fertility and light on AM fungal community.

Methods

We conducted a 3 year mesocosm experiment with a full two factorial design: light (full light or shade) and soil fertility (unfertilized or fertilized), on the Qinghai-Tibetan Plateau. Plant traits, soil characteristics and the AM fungal communities inside roots and in soils were measured.

Results

Shade reduced AM colonization of roots, fertilization reduced the hyphal abundance in the soil, and both factors reduced species richness of AM fungi inside plant roots. Fertilization exacerbated the negative impacts of shade on AM fungal abundance and diversity. We observed 15 phylotypes of AM fungi inside roots and ten morphotypes of AM fungal spores in the soil. Taxa responded differently to shade and fertilization and there was little congruence between the responses of fungi inside the roots and in the spore community.

Conclusions

Our findings indicate that both shade and fertilization reduce the abundance of AM fungi, but the two factors have different effects on the quality of plant roots as habitat for AM fungi.     

Bioavailability of zinc and phosphorus in calcareous soils as affected by citrate exudation

Aims

Zinc (Zn) and phosphorus (P) deficiency often occurs at the same time and limits crop production in many soils. It has been suggested that citrate root exudation is a response of plants to both deficiencies. We used white lupin (Lupinus albus L.) as a model plant to clarify if citrate exuded by roots could increase the bioavailability of Zn and P in calcareous soils.

Methods

White lupin was grown in nutrient solution and in two calcareous soils in a rhizobox. Rhizosphere soil solution was sampled to determine citrate, metals and P. Based on the measured citrate concentrations, a soil extraction experiment with citrate as extractant was done.

Results

Absence of Zn triggered neither cluster root formation nor citrate exudation of white lupin grown in nutrient solution, whereas low P supply did. The maximum citrate concentration (～1.5?mM) found in the cluster rhizosphere soil solution of one soil mobilized P, but not Zn. In the other soil the highest citrate concentration (～0.5?mM) mobilized both elements.

Conclusions

White lupin does not respond to low Zn bioavailability by increasing citrate exudation. Such a response was observed at low P supply only. Whether Zn and P can be mobilized by citrate is soil-dependent and the possible controlling mechanisms are discussed.     

Effects of arbuscular mycorrhizal inoculation on the growth,zinc distribution and photosynthesis of two citrus cultivars grown in low-zinc soil

Key message

‘Newhall’ and ‘Ponkan’ citrus cultivars grafted on trifoliate orange with mycorrhization by Glomus intraradices displayed different responses to low-Zn, and the optimal growth of ‘Newhall’ was more fungal inoculation dependent.

Abstract

The effects of arbuscular mycorrhizal (AM) fungus, Glomus intraradices, on plant growth, zinc (Zn) concentration and distribution, and photosynthesis were investigated in ‘Newhall’ navel orange (Citrus Sinensis) and ‘Ponkan’ tangerine (Citrus reticulata) grafted on the rootstock trifoliate orange (Poncirus trifoliata) exposed to low-Zn soil. Under the Zn-poor condition, the two cultivars showed similar decreases in growth, levels of leaf chlorophyll, gas exchange parameters, root soluble sugar, and Zn levels in various plant parts; however, the Zn percentage in the roots of ‘Ponkan’ was increased while in ‘Newhall’ it remained stable, indicating ‘Newhall’ may have a higher Zn translocation efficiency from the roots to the scion’s shoots. AM inoculation improved growth, Zn concentrations and photosynthesis in the two cultivars. AM-infected seedlings had lower minimal fluorescence values but higher maximal fluorescence values than non-infected seedlings. In addition, they were more efficient in terms of photosystem II’s (PS II’s) maximal and potential photochemistry. The greater changes appeared in ‘Newhall’, implying AM symbioses could alleviate the negative effects of low-Zn on the PS II reaction center. In addition, AM-infected ‘Newhall’ and ‘Ponkan’ seedlings had higher Zn percentages in the leaves but lower Zn ratios in the roots than non-AM-infected seedlings, especially in the former cultivar. These results indicate that G. intraradices has the potential to enhance the growth and Zn distribution in ‘Newhall’ grafted on trifoliate orange seedlings grown in low-Zn soil in a greenhouse.     

Mycorrhizal symbiosis affected by different genotypes of Pinus pinaster

Background and aims

Higher growth rate and morphological traits have been the major criteria for selecting trees in breeding programs. The symbiotic associations between P. pinaster and ectomycorrhizal fungi can be an effective approach to enhance plant development. The aim of this work was to assess whether the establishment of mycorrhizal symbiosis at nursery stage was affected by tree breeding.

Methods

Seeds of P. pinaster from a clonal population, designed to select for various traits, and from neighboring wild plants were inoculated with compatible ectomycorrhizal fungi: Suillus bovinus, Pisolithus tinctorius or Rhizopogon roseolus, and grown in individual cells containing forest soil, in a commercial forest nursery. Growth and nutritional traits, colonisation parameters and the fungal community established were assessed.

Results

R. roseolus and P. tinctorius were the most efficient isolates in promoting plant development. Inoculated selected saplings had an overall superior development than their wild counterparts, with up to a 4.9-fold in root dry weight and a 13.6-fold increase in the total number of ectomycorrhizal root tips. Differences in fungal community were revealed through the denaturing gradient gel electrophoresis profile of each treatment.

Conclusions

The results from our study suggest that the selected genotype benefits more from the mycorrhizal association and therefore this could be a valuable biotechnological tool for the nursery production of P. pinaster.     

Sample storage conditions alter colonisation structures of arbuscular mycorrhizal fungi and,particularly, fine root endophyte

Background and Aims

The structures of arbuscular mycorrhizal (AM) fungi (hyphae, arbuscules, vesicles, spores) are used to make inferences about fungal activity based on stored samples, yet the impact of storage method has not been quantified, despite known effects of temperature and host condition on AM fungal colonisation.

Methods

We measured how four storage treatments (cool or ambient conditions, with and without plant shoots attached, i.e. n?=?four treatment combinations) affected AM fungal colonisation of subterranean clover (Trifolium subterraneum L.) after 0, 2, 6 and 10 days of storage. Roots were assessed for colonisation of fine root endophyte and coarse AM fungi.

Results

For coarse AM fungi, total colonisation was unaffected, but arbuscules were reduced at Day 6 and increased again by Day 10, except Ambient-Minus-Shoots. There was a loss of vesicles in all treatments at Day 2, and an increase in spore number at Day 6 within Cool-Plus-Shoots. In contrast, for fine root endophyte, total colonisation was greatly reduced at Day 6 but increased again at Day 10, in all except the Cool-Plus-Shoots treatment.

Conclusions

Our data demonstrate that AM fungal activity is not suspended in commonly used plant storage conditions. Storage method and time impacted AM fungal colonisation, particularly for fine root endophyte. We recommend samples are processed within 2 days of harvest.

     

Effects of single and mixed inoculation with two arbuscular mycorrhizal fungi in two different levels of phosphorus supply on β-carotene concentrations in sweet potato (Ipomoea batatas L.) tubers

Aims

This study aimed to determine the effect of arbuscular mycorrhizal (AM) fungi and phosphorus (P) supply levels on β-carotene concentrations in sweet potato (Ipomoea batatas L.) tubers.

Methods

Two commercial AM fungal isolates of Glomus intraradices (IFP Glintra) and Glomus mosseae (IFP Glm) which differ in their life cycles were used. Sweet potato plants were grown in a horizontal split-root system that consisted of two root compartments. A root-free fungal compartment that allowed the quantification of mycelial development was inserted into each root compartment. The two root compartments were inoculated either with the same or with different AM isolates, or remained free of mycorrhizal propagules. Each fungal treatment was carried out in two P supply levels.

Results

In the low P supply level, mycorrhizal colonization significantly increased β-carotene concentrations in sweet potato tubers compared with the non-mycorrhizal plants. Glomus intraradices appeared to be more efficient in increasing β-carotene concentrations than G. mosseae. Dual inoculation of the root system with the two mycorrhizal fungi did not result in a higher increase in tuber β-carotene concentrations than inoculation with the single isolates. Improved P nutrition led to higher plant tuber biomass but was not associated with increased β-carotene concentrations.

Conclusions

The results indicate a remarkable potential of mycorrhizal fungi to improve β-carotene concentrations in sweet potato tubers in low P fertilized soils. These results also suggest that β-carotene metabolism in sweet potato tubers might be specifically activated by root mycorrhizal colonization.     

Phosphorus acquisition from phytate depends on efficient bacterial grazing, irrespective of the mycorrhizal status of Pinus pinaster

Background and aims

Phosphorus from phytate, although constituting the main proportion of organic soil P, is unavailable to plants. Despite the well-known effects of rhizosphere trophic relationships on N mineralization, no work has been done yet on P mineralization. We hypothesized that the interactions between phytate-mineralizing bacteria, mycorrhizal fungi and bacterial grazer nematodes are able to improve plant P use from phytate.

Methods

We tested this hypothesis by growing Pinus pinaster seedlings in agar containing phytate as P source. The plants, whether or not ectomycorrhizal with the basidiomycete Hebeloma cylindrosporum, were grown alone or with a phytase-producing bacteria Bacillus subtilis and two bacterial-feeder nematodes, Rhabditis sp. and Acrobeloides sp. The bacteria and the nematodes were isolated from ectomycorrhizal roots and soil from P. pinaster plantations.

Results

Only the grazing of bacteria by nematodes enhanced plant P accumulation. Although plants increased the density of phytase-producing bacteria, these bacteria alone did not improve plant P nutrition. The seedlings, whether ectomycorrhizal or not, displayed a low capacity to use P from phytate.

Conclusions

In this experiment, the bacteria locked up the phosphorus, which was delivered to plant only by bacterial grazers like nematodes. Our results open an alternative route for better utilization of poorly available organic P by plants.     

Prehistoric settlement activities changed soil pH, nutrient availability, and growth of contemporary crops in Central Europe

Background and aims

How prehistoric human settlement activities have changed soil chemical properties, plant nutrition and growth of contemporary crops is a question that has not been satisfactorily addressed. The aim of this paper was to study to what extent nutrient availability in the soil, together with nutrition and growth of spring barley (Hordeum vulgare), improved on sites of former sunken buildings (cropmarks) in comparison to their surroundings (controls) 1,700 years after abandonment of the buildings.

Methods

In the Czech Republic, a unique prehistoric settlement with many sunken buildings was discovered during aerial reconnaissance from cropmarks in stands of cereals. Soil and biomass samples were collected from cropmarks and controls in a barley crop in June 2012.

Results

A substantially higher content of organic matter, higher pH and concentrations of plant-available (Mehlich III) P, Ca, Mg, Cu and Zn were recorded in the sub-soil layer in cropmarks compared with controls, indicating the accumulation of wood ash and organic waste. In the arable layer, pH and concentrations of P, Ca and Mg were generally very high in both positions. Cropmarks were characterised by barley plants that were twice as tall as the controls, with significantly higher Ca, Mg and P concentrations.

Conclusions

Prehistoric settlement activity affected nutrient availability and plant growth in the previously settled area even after 1,700 years. We conclude that the chemical signature of prehistoric settlement activity can be detected from chemical analysis of the sub-soil layer as well as analysis of the contemporary arable layer and crop biomass.     

Root fungal colonisations of the understory grass <Emphasis Type="Italic">Deschampsia flexuosa</Emphasis> after top-canopy harvesting

Aims

Root fungal relationships in forest understory may be affected by tree harvesting. Deschampsia flexuosa forms a mutualistic symbiosis with arbuscular mycorrhizal (AM) fungi functioning in nutrient uptake, and a more loose association with dark septate endophytic (DSE) fungi. We asked how harvesting affects fungal colonisations and whether DSE is more prone to change than AM.

Methods

Deschampsia flexuosa plants were sampled close to a control or a cut tree after top-canopy harvesting in a primary successional site. Colonisations were studied using light microscopy. Shoot N%, vegetation cover and soil nutrients were determined.

Results

Tree harvesting did not affect vegetation and soil parameters, except potassium (K⁺) increasing near cut trees. AM colonisation did not change, while DSE increased. Shoot N% increased with increasing DSE near cut trees. Hyaline septate (HSE) hyphae and soil K⁺ and magnesium (Mg²⁺) were positively correlated near control trees. Lichen cover and HSE correlated negatively.

Conclusions

DSE colonisation increased but AM did not change after harvesting. Positive correlation of DSE with shoot N% near cut trees may suggest a role for DSE in favouring plant nitrogen uptake after disturbance in an open microsite. HSE may play a role in K⁺ and Mg²⁺ uptake.

     

Root morphology acclimation to phosphorus supply by six cultivars of <Emphasis Type="Italic">Trifolium subterraneum</Emphasis> L

Aims

Trifolium subterraneum L. is the predominant annual pasture legume in southern Australia. Cultivars with improved phosphorus (P) foraging ability would improve the P-use efficiency of agricultural systems. We therefore investigated variation in root traits related to P-uptake among six cultivars.

Methods

Micro-swards were grown at six levels of P in field soil with indigenous arbuscular mycorrhizal (AM) fungi for six weeks. Dry matter yield, tissue P concentration, rhizosphere carboxylates, AM fungal colonisation and root morphological traits were measured.

Results

The cultivars showed similar shoot and root yield responses to P supply. Average root diameter did not change, specific root length (SRL) increased and root tissue density (RTD) decreased with increased P supply. Amounts of total rhizosphere carboxylates were low (<1.2 nmol cm^?1 root). The percentage of root length colonised by AM fungi was greatest (29–43 %) at an intermediate level (8 mg kg^?1 dry soil) of P supply.

Conclusions

Most differences among cultivars were reasonably consistent across P supply levels, indicating greater numbers of lines could be screened reliably at a single P level. Low colonisation by AM fungi at low P supply deserves consideration when selecting soils for cultivar comparisons. Increased SRL and decreased RTD at high P supply likely result from self-shading within the micro-swards and warrant further investigation.

     

Biofortification of zinc in wheat grain by the application of sewage sludge

Background and aims

Increasing the concentrations of the essential micronutrient Zn in staple crops like grain is desirable for human nutrition. We investigated the long-term ability of municipal treatment works sewage sludge, liquid sewage sludge and ZnCO₃ applied to soils to increase Zn in in wheat grain (Triticum aestivum L.) in a number of field experiments conducted on different soils.

Methods

We used six long-term field experiments that were set up on contrasting soils in England and the target applications were built up between 1994 and 1997. Topsoil samples and harvested grain samples were taken and air dried in 1999, 2001, 2003 and 2005. Relationships between grain Zn concentrations and soil properties and changes with time were examined.

Results

Wheat grain Zn concentrations increased with soil Zn concentrations in a similar log-log relationship with all of the Zn sources tested. Comparing total or extractable Zn in soil as explanatory factors showed little benefit of using extractable Zn measurements to predict grain concentrations over total Zn. Additional factors such as soil pH or organic carbon did not explain much more of the variation in grain Zn in our experiments. However, grain Zn concentrations did not respond at all at a site with pH 7.7.

Conclusions

Sewage sludge applications to soil can increase grain Zn concentrations for at least 2 to 8?years after application and has similar effectiveness to ZnCO₃.     

Net root growth and nutrient acquisition in response to predicted climate change in two contrasting heathland species

Background and aims

Accurate predictions of nutrient acquisition by plant roots and mycorrhizas are critical in modelling plant responses to climate change.

Methods

We conducted a field experiment with the aim to investigate root nutrient uptake in a future climate and studied root production by ingrowth cores, mycorrhizal colonization, and fine root N and P uptake by root assay of Deschampsia flexuosa and Calluna vulgaris.

Results

Net root growth increased under elevated CO₂, warming and drought, with additive effects among the factors. Arbuscular mycorrhizal colonization increased in response to elevated CO₂, while ericoid mycorrhizal colonization was unchanged. The uptake of N and P was not increased proportionally with root growth after 5 years of treatment.

Conclusions

While aboveground biomass was unchanged, the root growth was increased under elevated CO₂. The results suggest that plant production may be limited by N (but not P) when exposed to elevated CO₂. The species-specific response to the treatments suggests different sensitivity to global change factors, which could result in changed plant competitive interactions and belowground nutrient pool sizes in response to future climate change.     

As,Pb, Sb,and Zn transfer from soil to root of wild rosemary: do native symbionts matter?

Background and aims

This is an in natura study aimed to determine the potential of Rosmarinus officinalis for phytostabilization of trace metal and metalloid (TMM)-contaminated soils in the Calanques National Park (Marseille, southeast of France). The link between rosemary tolerance/accumulation of As, Pb, Sb, and Zn and root symbioses with arbuscular mycorrhizal (AM) fungi and/or dark septate endophytes (DSE) was examined.

Methods

Eight sites along a gradient of contamination were selected for soil and root collections. TMM concentrations were analyzed in all the samples and root symbioses were observed. Moreover, in the roots of various diameters collected in the most contaminated site, X-ray microfluorescence methods were used to determine TMM localization in tissues.

Results

Rosemary accumulated, in its roots, the most labile TMM fraction in the soil. The positive linear correlation between TMM concentrations in soil and endophyte root colonization rates suggests the involvement of AM fungi and DSE in rosemary tolerance to TMM. Moreover, a typical TMM localization in root peripheral tissues of thin roots containing endophytes forming AM and DSE development was observed using X-ray microfluorescence.

Conclusions

Rosemary and its root symbioses appeared as a potential candidate for a phytostabilization process of metal-contaminated soils in Mediterranean area.