Pisolithus Tinctorius Research Articles

Antioxidants in Pinus pinaster roots and mycorrhizal fungi during the early steps of symbiosis

Ectomycorrhizal symbiosis between fungi mycelia and the roots of some plants could have important effects in the levels of antioxidants of both partners. In the present work, the effects of co-culture period in the antioxidant properties and antioxidants (phenolics, tocopherols and sugars) production during early steps of in vitro mycorrhization (Pinus pinaster–Pisolithus arhizus and Pinus pinaster–Paxillus involutus) were evaluated. The studied parameters were determined in each culture element (root, mycelium and medium) in order to understand the response of each partner to the symbiotic association. P. arhizus proved to be more compatible with P. pinaster than P. involutus, since the antioxidant activity in the latter species increased from 48 to 72h, while P. arhizus antioxidant properties decreased at the same period (indicative of less oxidative stress). Despite P. involutus proved to be less suitable to be included in forestation programs using mycorrhization processes, it revealed a higher potential for bioactive compounds production in the early steps of symbiosis. A maximal value of phenolics content was obtained after the first 6h.

The full length <i>PtSRP</i> (<i>Pisolithus tinctorius</i> symbiosis related protein) fungal mRNA encodes a potential marker of ectomycorrhiza formation

The Pisolithus tinctorius symbiosis related protein expressed sequence tag (EST PtSRP) was previously identified in the first hours of the interaction between the fungus Pisolithus tinctorius and sweet chestnut Castanea sativa, and partially characterized as a fungal marker gene of ectomycorrhizal symbiosis formation. We used the 5’ rapid amplification of cDNA ends (RACE) to obtain the PtSRP mRNA 5’ region, and together with our previously reported 3’ mRNA region, the full mRNA sequence was assembled by use of bioinformatics tools and deposited to GenBank (Accession: GU733439). The full-length mRNA sequence (636 bp) revealed the locations of the 5’ and 3’ untranslated regions (UTRs) and contained the Kozak sequence (ccc aag ATG A) in the 5’ UTR. The in silico translated PtSRP open reading frame (ORF) codes for a 127 amino acid protein and contained four putative post-translational modification sites (two N-glycosylation and two phosphorylation). The protein secondary structure is postulated to be composed of one N-terminal hydrophobic transmembrane alpha helix and at least six hydrophilic beta-strands spread across the protein. Sub-cellular localization prediction suggests that the protein is involved in cellular secretory pathway, supported by the presence of a cleavage site motif close to the membrane anchor. The data presented herein indicate the role of PtSRP as a fungal membrane secreted protein involved in early stages of ectomycorrhizal formation, with application as a possible marker for nascent ectomy-corrhiza fungal development.

Detection of Forest Ectomycorrhizal Fungi- Pisolithus tinctorius by Nested PCR

For investigating a suitable molecular approach to detect ectomycorrhizal fungi(EMF),the internal transcribed spacer(ITS) region of the EMF Pisolithus tinctorius was amplified with universal primers ITS1-F/ITS4-B and sequenced.Based on the conserved sequence determined by alignment using Clustal W,a pair of specific primers named PtF/PtR were designed by Primer premier 5.0.The specificity of the primer pair was then analyzed in 10 P.tinctorius strains together with 6 reference strains,and a 347 bp product was amplified from all P.tinctorius strains but there was no product from the 6 reference strains.A nested PCR procedure using ITS1-F/ITS4-B as the first-round primers followed by PtF/PtR increased detection sensitivity 1 000-fold to 10 fg DNA template.The target EMF from the mycorrhiza of Pinus massoniana potted seedlings inoculated with P.tinctorius was detected by this technique.The results indicated that the nested PCR assay could be used as a sensitive and accurate way to detect forest EMF P.tinctorius.

Erratum to: Tolerance to and Accumulation of Cadmium by the Mycelium of the Fungi Scleroderma citrinum and Pisolithus tinctorius

Erratum to: Tolerance to and Accumulation of Cadmium by the Mycelium of the Fungi Scleroderma citrinum and Pisolithus tinctorius

Effects of different ectomycorrhizal fungi on somatic embryogenesis of Abies cephalonica Loud

The effects of ectomycorrhizal (ECM) fungi, including Laccariabicolor (Maire) Orton, Laccaria laccata (Scop., Fr.) Berk. and Br., along with two strains of Pisolithus tinctorius (Pers.) Coker and Couch, on the proliferation and subsequent maturation of two embryogenic cell lines of Abies cephalonica Loud., designated lines 6 and 8, were investigated. In the presence of these ECM fungi, the proliferation of both embryogenic cell lines was inhibited. L. bicolor and P. tinctorius strain 2 resulted in the highest inhibition rates. On the other hand, cultivation of embryogenic cultures along with ECM fungi, termed a dual culture, increased radial growth of both P. tinctorius strains; whereas, L. bicolor and L. laccata did not grow as well in the presence of embryogenic cell masses. The dual culture during the proliferation period of embryogenic cells, however, enhanced the subsequent embryo formation and maturation of A. cephalonica; i.e. the capability of embryogenic cell lines to form somatic embryos as well as increasing the mean number of somatic embryos per 1 g fresh weight of embryogenic cell mass. However, levels of responses were highly dependent on the interaction between the specific embryogenic cell line and fungal strain.

Defence responses of Norway spruce seedlings to elicitors of ectomycorrhizal fungus Pisolithus tinctorius and pathogen Heterobasidion annosum are affected by zeatin riboside

Cytokinins are known to attenuate defence responses of plants after elicitor application or inoculation with fungi. To evaluate their role in the regulation of colonisation of Norway spruce (Picea abies) seedlings with mycorrhizal and pathogenic fungus, we analysed the effects of zeatin riboside (ZR) on: i) growth of ectomycorrhizal fungus Pisolithus tinctorius and pathogen Heterobasidion annosum in axenic cultures, ii) colonisation intensity of selected fungi on P. abies seedlings and iii) induction of defence reactions of spruce seedlings following elicitor treatment. Mycorrhizal fungus P. tinctorius showed increased growth at concentrations higherthan 10-2 µM ZR that was accompanied by increased ergosterol concentrations. In contrast, decreased growth of pathogen H. annosum was observed at the highest ZR (10 µM) concentration. ZR treatment also increased colonisation of spruce seedlingswith the mycorrhizal fungus. Application of cell wall preparations of both fungi increased peroxidase (POD) activity in the roots of treated spruce seedlings, whereas only elicitors of H. annosum increased also phenylalanine ammonia-lyase (PAL) activity,levels of soluble phenolics and salicylic acis (SA) concentrations. Application of ZR negated the increased activity of POD that was observed in elicitor treated seedlings, accompanied by increased levels of soluble phenolics in the roots of elicitated seedlings. In contrast, no effects of ZR treatment on PAL activity and SA accumulation could be observed. Our results suggest involvement of ZR in the regulation of cell wall modifications during the fungal colonisation with P. tinctorius and formation of ectomycorrhizae, by affecting the growth of fungal partner and non-specific defence reactions of the plant host.

Effects of the ectomycorrhizal fungus Pisolithus tinctorius and Cd on physiological properties and Cd uptake by hybrid poplar Populus alba × glandulosa

The effects of the ectomycorrhizal fungus Pisolithus tinctorius and cadmium (Cd) on physiological properties and Cd uptake by Populus alba <TEX>${\times}$</TEX> glandulosa was investigated under greenhouse conditions. Cd treatment decreased the photosynthetic rate (<TEX>$P_N$</TEX>) of both non-mycorrhizal (NM) plants (16.3%) and ectomycorrhizal (ECM) plants (11.5%). In addition, the reduction in total dry weight by Cd treatment was greater in ECM plants (24.3%) than that in NM plants (17.6%). Mycorrhizal infection increased the <TEX>$P_N$</TEX> and transpiration rate in both control and Cd-treated plants. Cd treatment increased superoxide dismutase (SOD) activity and decreased glutathione reductase activity, and the increase of SOD activity by Cd treatment was greater in NM plants (40.3%) than that in ECM plants (3.7%). Thiol content increased in both NM and ECM plants treated with Cd solution, and the increase in thiol content in NM plants (43.9%) was greater than that of ECM plants (15.6%). Cd uptake in the leaves, stems, and roots of ECM plants was 69.9%, 167.2% and 72.8%, respectively, higher than in the NM plants. However, the increase in Cd uptake ability of ECM plants resulted in a reduction in dry weight.

Tolerance to and Accumulation of Cadmium by the Mycelium of the Fungi Scleroderma citrinum and Pisolithus tinctorius

The behavior of ectomycorrhizal (ECM) fungi on exposure to cadmium dependent upon isolation remains a poorly understood phenomenon. The in vitro growth, tolerance, and accumulation of Cd were studied in three strains of ECM fungi exposed to six Cd concentrations (0-10 mg L(-1)). The fungi studied were a strain of Scleroderma citrinum Persoon (Sc) isolated from a tailings heap containing 5 mg kg(-1) available Cd, and two strains of Pisolithus tinctorius (Pers.) Coker and Couch from unpolluted sites (Pt1 and Pt2), both common ECM fungi used for remediation. The growth kinetic (36 days) of Sc was not affected by Cd concentration. By contrast, the ED(50) in Pt1 and Pt2 occurred at 4.8 and 6.9 mg L(-1) of Cd, respectively. The biomass of the three fungi exposed to the highest Cd concentration (10 mg L(-1)) was significantly different. Sc presented the highest biomass, while this was strongly reduced for Pt1 and Pt2. The tolerance index for Sc ranged from 78% to 95% at all Cd concentrations tested, while for Pt1 it was 49% and 31%, and for Pt2 it was 62% and 35% at 5 and 10 mg of Cd L(-1), respectively. The mycelium of both Pt strains accumulated more Cd than the Sc mycelium. At the highest Cd concentration, Pt1 and Pt2 accumulated 1.9 and 1.7 times more Cd than Sc. This study suggests that regardless of the differences in tolerance to Cd by the three ECM fungi, they could have biotechnological applications for soil remediation. However, Sc has greater possibilities of being used successfully when high concentrations of Cd prevail in the environment.

Effect of the mycorrhizal symbiosis time in the antioxidant activity of fungi and Pinus pinaster roots, stems and leaves

The ectomycorrhizal (ECM) symbiosis that develops between the roots of host trees and the soil ECM fungi is an important factor towards the survival, health and growth of these trees, as it stimulates their water and nutrient uptake. Ectomycorrhizal colonization can result in the deposition of phenolic compounds in peripheral cortex cells and a similar answer can be recognized as one way of plant defense against pathogenic infections. The aim of the present work was the evaluation of antioxidant potential of the ectomycorrhizal fungi, Paxillus involutus and Pisolithus arhizus, in presence and absence of the symbiont – Pinus pinaster – in response to the symbiotic association, under different contact periods (45 days and 48 h). Phenolic contents in mycelia, culture media and plant leaves, stems and roots were determined by Folin–Ciocalteu assay, and their antioxidant properties were evaluated by three in vitro assays: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, reducing power and lipid peroxidation inhibition through β-carotene bleaching inhibition. Comparing the response of the symbionts to the host species, P. pinaster benefited most with the association, because it generally decreases the phenolics content and the antioxidant activity values in the presence of both ectomycorrhizal fungi along time. P. arhizus also decreased, in general, its phenolics levels and antioxidant properties, alongside with P. pinaster, unlike P. involutus that did not have the same response as it increases its content of phenolics and some of its values of antioxidant activity. These results can be considered as an hypothetical signal of a symbiotic differential compatibility of mycorrhizal fungi for a host.

Intraspecific variability of Pisolithus spp. as a response to changes in soil characteristics in a Moroccan cork oak plantation

Soil characteristics may be an important factor in structuring fungal communities. We relate the above- and below-ground distribution of the ectomycorrhizal fungi (EMF) Pisolithus spp. to changes in soil chemistry in a Quercus suber plantation located in the Maâmora forest (Morocco). Intrageneric variability of 115 fruit bodies was studied, using morphological characterization, restriction fragment length polymorphism (RFLP) analyses and internal transcribed spacer (ITS) sequencing. Two Pisolithus spp. genotypes were identified: 97% of the fruit bodies represent Pisolithus arhizus; the remaining 3% correspond to Pisolithus species 4, as previously described by Martin et al. [New Phytologist 153 (2002) 345-357]. Multivariate analysis (PCA) showed that species structure was strongly correlated with soil characteristics. P. arhizus fruit bodies were frequently found in the Eastern part of the plot at low molasic clay, N and P contents, while Pisolithus species 4 were collected only in the Western part at the high molasic clay, N and P contents. To determine whether this change in fruit body structure was expressed at the belowground level, we used morphological and molecular techniques to characterize the Pisolithus ectomycorrhizas. Only ectomycorrhizas of P. arhizus were identified at the low P–N–molasic clay site and disappeared completely at the high P–N–red clay site, where no Pisolithus species 4 mycorrhizas were found. However, autocorrelation among soil parameters makes it difficult to isolate the effects of individual parameters. These results show the local-scale impact of natural spatial heterogeneity on an ectomycorrhizal fungal genus.

Reforestation of burned stands: The effect of ectomycorrhizal fungi on Pinus pinaster establishment

The area occupied by Pinus pinaster in Portugal is rapidly diminishing because of forest fires. Ectomycorrhizal fungi form obligate, mutually beneficial associations with P. pinaster which improve plant growth and resistance to adverse conditions. The aim of this work was to assess whether native ectomycorrhizal fungi could be a useful tool in the reforestation of burned areas. The work was conducted in a forest nursery greenhouse, where P. pinaster seedlings were inoculated with compatible ectomycorrhizal fungal isolates: Suillus bovinus, Pisolithus tinctorius, Rhizopogon roseolus, and a mixture of the three fungi, using burned and unburned forest soil as substrate. Inoculation significantly enhanced the growth of P. pinaster, with R. roseolus proving to be the most effective in burned soil, with an 8-fold increase in plant fresh weight. Overall, inoculation stimulated growth most in burned than in unburned soil. This study suggests that inoculation with selected ectomycorrhizal fungi in containerised nurseries can be an advantageous approach for the successful establishment of P. pinaster in burned soil. The obtained results point out to the interest of extending these studies into fire-impacted areas, using ectomycorrhizal fungi as a biological tool.

Mycorrhizal induction of phenolic compounds and antioxidant properties of fungi and seedlings during the early steps of symbiosis

Ectomycorrhizal (ECM) symbiosis plays a major role in nutrient cycling and the functioning of forest ecosystems. Trees with well-developed ectomycorrhizas are more resistant to environmental stresses such as drought and to biotic stresses such as root pathogens. The establishment of ECM symbiosis is triggered by signals produced by both partners. These signals lead to morphological changes and a complex development of specific structures in both the plant root and the fungus. The development of the ECM fungi, Paxillus involutus and Pisolithus arhizus, in the presence and the absence of the symbiont—Pinus pinaster—was evaluated as well as their antioxidant properties and phenolic compounds composition in response to the symbiotic association. ECM fungi grew less in the presence of P. pinaster, with P. arhizus being less affected in growth and thus being more adapted to this association. Protocatechuic acid was found only in P. involutus, while P. pinaster roots, both in association and isolated, proved to have other phenolic acids, such as p-hydroxybenzoic and p-coumaric acids. The symbiosis between P. involutus and P. pinaster had no major effects on the symbionts, while the association between P. arhizus and P. pinaster seems to generally decrease the antioxidant effects of both symbionts, despite the increase in p-coumaric and cinnamic acids in the ECM fungi.

Effect of competitive interactions between ectomycorrhizal and saprotrophic fungi on Castanea sativa performance

In Northeast of Portugal, the macrofungal community associated to chestnut tree (Castanea sativa Mill.) is rich and diversified. Among fungal species, the ectomycorrhizal Pisolithus tinctorius and the saprotroph Hypholoma fasciculare are common in this habitat. The aim of the present work was to assess the effect of the interaction between both fungi on growth, nutritional status, and physiology of C. sativa seedlings. In pot experiments, C. sativa seedlings were inoculated with P. tinctorius and H. fasciculare individually or in combination. Inoculation with P. tinctorius stimulated the plant growth and resulted in increased foliar-N, foliar-P, and photosynthetic pigment contents. These effects were suppressed when H. fasciculare was simultaneously applied with P. tinctorius. This result could be related to the inhibition of ectomycorrhizal fungus root colonization as a result of antagonism or to the competition for nutrient sources. If chestnut seedlings have been previously inoculated with P. tinctorius, the subsequent inoculation of H. fasciculare 30 days later did not affect root colonization, and mycorrhization benefits were observed. This work confirms an antagonistic interaction between ectomycorrhizal and saprotrophic fungi with consequences on the ectomycorrhizal host physiology. Although P. tinctorius is effective in promoting growth of host trees by establishing mycorrhizae, in the presence of other fungi, it may not always be able to interact with host roots due to an inability to compete with certain fungi.

New Report of the Mycorrhizal Association betweenPisolithus tinctorius(Sclerodermataceae, Basidiomycota) andQuercus coccifera(Fagaceae, Angiospermae)

Abstract In this paper we described for the first time the mycorrhiza between the fungus Pisolithus tinctorius and the kermes oak Quercus coccifera, obtained by inoculation. The synthesis was performed under nursery conditions using vegetative (40 mL/plant) or sporal (108 spores/plant) inoculum of P. tinctorius basidiomata collected from two different locations and putative hosts, and containers filled with a sterile mixture of sphagnum peat/ black peat/perlite+vermiculite. The mycorrhizal association is described and illustrated in detail and compared with other known P. tinctorius mycorrhiza. Mycorrhizae were monopodial-pinnate to pyramidal pinnate, cream to brown colour, mantle surface loosly woven, extramatrical mycelium abundant, rhizomorophs scarce and well differentiated, sclerotia frequent, subglobose to lemon-shaped, mantle plectenchymatous with three layers. The conditions of inoculation are also discussed. A beneficial effect of the mycorrhizal symbiosis on plant growth attributes is suggested.

Testing the effect of routine fungicide application on ectomycorrhiza formation on Pinus halepensis seedlings in a nursery

SummaryThe fungicides benomyl, captan, hymexazol, iprodione, propamocarb hydrochloride and thiram were applied in a combined schedule to Pinus halepensis seedlings grown under nursery conditions to study their effect on mycorrhiza formation and plant growth. The inoculated fungi Pisolithus tinctorius (strains 3SR and Mx) and Lactarius deliciosus formed mycorrhizas when fungicides were applied. However, fungicide application led to a significant decrease of mycorrhizal plants with Pisolithus tinctorius strain 3SR (from 62% without fungicides to 35% with fungicides). This effect could be attributed to a direct effect of fungicides on mycelial growth. The morphometric and nutritional parameters were very similar in plants treated with or without fungicides, and no differences on sanitary status of plants were noted. The use of preventive fungicide treatments in plant production, which may have a harmful effect on the environment, is therefore questioned.

A comparative study of tocopherols composition and antioxidant properties of in vivo and in vitro ectomycorrhizal fungi

In aerobic organisms, the free radicals are constantly being produced during the normal cellular metabolism. The antioxidant properties of many organisms and particularly of wild mushrooms with their content in antioxidant compounds such as tocopherols, can detoxify potentially damaging forms of activated oxygen. Herein, a comparative study of tocopherols composition and antioxidant properties of in vivo (fruiting bodies) and in vitro (mycelia) ectomycorrhizal fungi: Paxillus involutus and Pisolithus arhizus. Tocopherols were determined by high performance liquid chromatography (HPLC) coupled to a fluorescence detector. The antioxidant properties were studied in terms of DPPH radical-scavenging activity, reducing power and inhibition of β-carotene bleaching. Fruiting bodies revealed the highest antioxidant properties, including scavenging effects on free radicals (EC 50 = 0.61 and 0.56 mg/ml) and inhibition of lipid peroxidation capacity (EC 50 = 0.40 and 0.24 mg/ml for P. involutus and P. arhizus, respectively), than mycelia produced in vitro cultures. Nevertheless, mycelia revealed higher levels of total tocopherols than fruiting bodies, and particularly P. arhizus mycelium proved to be a powerful source of γ-tocopherol (154.39 μg/g dry weight).

Inhibitory effect of pisosterol on human glioblastoma cell lines with C‐MYC amplification

Despite the remarkable progress in the characterization of the molecular pathogenesis of glioblastoma multiforme (GBM), these tumors remain incurable and, in most cases, refractory to aggressive cytotoxic treatments. We conducted a morphological and cytogenetic study in two GBM cell lines (U343 and AHOL1), before and after treatment with pisosterol (at 0.5, 1.0 and 1.8 µg ml⁻¹), a triterpene isolated from the fungus Pisolithus tinctorius. No significant alteration was observed in the morphology and frequency of chromosomal abnormalities in the cell lines analyzed after treatment with pisosterol. Using fluorescence in situ hybridization analysis with a locus-specific probe for C-MYC showed that 72% of U343 and 65% of AHOL1 cells contained more than two alleles of C-MYC before treatment. After treatment, no effects were detected at lower concentrations of pisosterol (0.5 and 1.0 µg ml⁻¹). However, at 1.8 µg ml⁻¹ of pisosterol, only 33% of U343 cells and 15% of AHOL1 cells presented more than two fluorescent signals, suggesting that pisosterol blocks the cells with gene amplification. Cells that do not show a high degree of C-MYC gene amplification have a less aggressive and invasive behavior and are easy targets for chemotherapy. Therefore, further studies are needed to examine the use of pisosterol in combination with conventional anti-cancer therapy.

Ectomycorrhizal fungi as an alternative to the use of chemical fertilisers in nursery production of Pinus pinaster

Addition of fertilisers is a common practice in nursery production of conifer seedlings. The aim of this study was to evaluate whether ectomycorrhizal (ECM) fungi can be an alternative to the use of chemical fertilisers in the nursery production of Pinus pinaster. A greenhouse nursery experiment was conducted by inoculating seedlings obtained from seeds of P. pinaster plus trees with a range of compatible ECM fungi: (1) Thelephora terrestris, (2) Rhizopogon vulgaris, (3) a mixture of Pisolithus tinctorius and Scleroderma citrinum, and (4) a mixture of Suillus bovinus, Laccaria laccata and Lactarius deterrimus, using forest soil as substrate. Plant development was assessed at two levels of N-P-K fertiliser (0 or 600 mg/seedling). Inoculation with a mixture of mycelium from S. bovinus, L. laccata and L. deterrimus and with a mixture of spores of P. tinctorius and S. citrinum improved plant growth and nutrition, without the need of fertiliser. Results indicate that selected ECM fungi can be a beneficial biotechnological tool in nursery production of P. pinaster.

Fraxinus–Glomus–Pisolithus symbiosis: Plant growth and soil aggregation effects

Abstract It has been established that arbuscular mycorrhizal (AM) fungi are involved in the conservation of soil structure. However, the effect of ectomycorrhizal (EM) fungi alone or in interaction with AM fungi in soil structure has been much less studied. This experiment evaluated EM and AM fungi effects on soil aggregation and plant growth. Ash plants ( Fraxinus uhdei ) were grown in pots, and were inoculated with Glomus intraradices and Pisolithus tinctorius separately but also in combination. Our results showed that F. uhdei established a symbiotic association with EM and AM fungi, and that these organisms, when interacting, showed synergistic and additive effects on plant growth compared to singly inoculated treatments. EM and AM fungi prompted changes in root morphology and increased water-stable aggregates. AM fungi affect mainly small-sized macroaggregates, while EM and EM–AM fungi interaction mainly affected aggregates bigger than 0.5 mm diameter. These results suggest that ectomyccorrhizal as well as arbuscular mycorrhizal fungi should be considered in restoration programs with Fraxinus plants.

MYCORRHIZAL INOCULATION OF CHESTNUT SEEDLINGS: EFFECT ON SURVIVAL AND GROWTH AFTER TRANSPLANTATION

Trees of several species cannot develop without mycorrhizas. The first trials of controlled mycorrhization were made in Australia early in the twentieth century. Mycorrhizal inoculation is based on the main findings: (1) any mycorrhiza is better than none for plants produced in greenhouses/nurseries; (2) some species are more beneficial than others under specific environmental conditions. Building on mycorrhizal studies on chestnut (Castanea sativa Mill.) inoculated plants, work was started on inoculum production in order to inoculate chestnut plants in greenhouse/nursery and evaluate its effects. A consortium programme was started with the production of inoculum and mycorrhization of seedlings in greenhouse, followed by transference to nurseries. We report the methodology and results obtained in three year long trials with chestnut tree seedlings mycorrhization with Pisolithus tinctorius. Mycorrhizal effects were analyzed through germination rates, survival and growth along the germination, and weaning processes. Inoculation of the germination substrate did not influence germination rates, bpt affected plant growth: mycorrhizal plants are thicker than non-mycorrhizal ones. Inoculation of the substrate at sowing followed by previous acclimation revealed more favourable for plant survival and growth than inoculation and direct transfer to the nursery. When in contact with soils infected by Phytophthora cinnamomi mycorrhizal plants revealed higher survival capacity than non-mycorrhizal plants although this survival is dependent on the time of mycorrhization.