Cultured arbuscular mycorrhizal fungi and native soil inocula improve seedling development of two pioneer trees in the Andean region

The tree species Alnus acuminata and Morella pubescens, native to South America, are candidates for soil quality improvement and afforestation of degraded areas and may serve as nurse trees for later inter-planting of other trees, including native crop trees. Both species not only form symbioses with arbuscular mycorrhizal fungi (AMF) and ectomycorrhizal fungi (EMF), but also with N₂-fixing actinobacteria. Because tree seedlings inoculated with appropriate mycorrhizal fungi in the nursery resist transplanting stress better than non-mycorrhizal seedlings, we evaluated for A. acuminata and M. pubescens the potential of inoculation with mycorrhizal fungi for obtaining robust tree seedlings. For the first time, a laboratory-produced mixed AMF inoculum was tested in comparison with native soil from stands of both tree species, which contains AMF and EMF. Seedlings of both tree species reacted positively to both types of inocula and showed an increase in height, root collar diameter and above- and belowground biomass production, although mycorrhizal root colonization was rather low in M. pubescens. After 6 months, biomass was significantly higher for all mycorrhizal treatments when compared to control treatments, whereas aboveground biomass was approximately doubled for most treatments. To test whether mycorrhiza formation positively influences plant performance under reduced water supply the experiment was conducted under two irrigation regimes. There was no strong response to different levels of watering. Overall, application of native soil inoculum improved growth most. It contained sufficient AMF propagules but potentially also other soil microorganisms that synergistically enhance plant growth performance. However, the AMF inoculum pot-produced under controlled conditions was an efficient alternative for better management of A. acuminata and M. pubescens in the nursery, which in the future may be combined with defined EMF and Frankia inocula for improved management practices.     

Response of <Emphasis Type="Italic">Theobroma cacao</Emphasis> and <Emphasis Type="Italic">Inga edulis</Emphasis> seedlings to cross-inoculated populations of arbuscular mycorrhizal fungi

Response of Theobroma cacao L. (cacao) and the shade tree Inga edulis Mart. (inga) seedlings from an organically-grown cacao plantation to inoculation with native arbuscular mycorrhizae forming fungi (AMF) was studied in a cross-inoculation assay under greenhouse conditions. Seedlings of inga and cacao were grown in pots filled with heat-treated soil from the plantation. Control was heat-treated soil without inoculum and roots of cacao and inga from the plantation were applied as AMF inocula. Undisturbed soil blocks were used as a “positive control” of the inoculation potential of untreated soil and roots combined. No AMF structures were observed in the roots of either species in the heat-treated control. All inocula were infective in both hosts and the differences in the total AMF colonization percentage between the hosts were not significant but inga had significantly higher colonization by hyphal coils and arbuscules. Cacao roots but neither inga roots nor soil block inocula stimulated cacao growth. All inocula significantly increased growth of inga, which had higher relative mycorrhizal responsiveness than cacao. Thus, in spite of the strong infectivity of the inocula in both hosts, cacao and inga responded differently to the same AMF populations. The strong conspecific preference of cacao suggests that attention must be paid to the AMF inoculum used for this species. However, the strong response of inga to cacao root inoculum indicates that the two species may share same AMF symbionts, thus enabling positive interactions between them, including formation of common mycelial networks.     

Arbuscular mycorrhizas and performance of Polylepis australis trees in relation to livestock density

Polylepis australis trees endemic to Argentina dominate the canopy of subtropical high altitude forests. Here, livestock rearing is the main economic activity and is suspect of the low performance of P. australis trees through direct and indirect effects which could include the reduction in arbuscular mycorrhizal fungi (AMF) and their benefit to trees. To elucidate the role of AMF, we compare plant performance indicators, arbuscular mycorrhizal (AM) colonization and AMF communities in 20 trees distributed in two areas of central Argentina which differed in livestock grazing intensity. The area with high livestock density presented more soil degradation and trees with a lower overall plant performance than the area with reduced livestock density. The AM colonization values of P. australis were considerably higher than reported for other tree species and the area with high livestock density had a lower proportion of arbuscules and higher proportion of hyphae, while vesicles and AM colonization – all structure considered together – did not differ between areas. Overall AMF spore number and of most species when considered separately was significantly higher in the area with high livestock density, suggesting a high tolerance and adaptation of AMF to livestock. We conclude that a reduction in livestock improves the performance of P. australis, that this improvement could be mediated by an increase in the proportion of arbuscules, but there does not appear to be any limitation in AM colonization or AMF spore number which could otherwise be limiting forest restoration.     

Growth response of three native timber species to soils with different arbuscular mycorrhizal inoculum potentials in South Cameroon: Indigenous inoculum and effect of addition of grass inoculum

After tropical forest disturbance, mycorrhizal inoculum could be insufficient. Increasing mycorrhizal density through inoculum addition is then crucial for successful regeneration of deforested lands. Greenhouse bioassays were set up to determine the effectiveness of native arbuscular mycorrhizal fungi in soils from different disturbance stages on the growth of three important timber species, Terminalia superba, Distemonanthus benthamianus, and Entandrophragma utile. Soils were collected from late and early successional forest stands, fields of food crops, fallow of Chromolaena odorata, skid trails, bare soil landings, and landings with the pioneer tree Musanga cecropioides. These soils were used to grow seedlings without or with addition of an inoculum collected under the grass Paspalum conjugatum. The extent to which seedlings responded to indigenous inoculum and inoculum addition varied with tree species and with mycorrhizal inoculum potential. After inoculum addition, Terminalia strongly increased root colonization with a small increase in shoot dry weight and Distemonanthus hardly increased root colonization but showed a strong increase in shoot dry weight. Entandrophragma increased both root colonization and shoot dry weight. Plant biomass was lower in soils with low inoculum potential such as late successional stands, skid trails, and both kinds of landings; the mycorrhizal inoculation effect was then large. Plant biomass was high in agricultural fields and fallow; mycorrhizal inoculation effect was sometimes even negative. These data indicate that low inoculum might limit plant re-establishment after disturbance and that mycorrhizal inoculation has a potential for improving seedling establishment on deforested land.     

Response of Acacia senegal (L.) Willd. to inoculation with arbuscular mycorrhizal fungi isolates in sterilized and unsterilized soils in Senegal

The benefits of inoculation with six arbuscular mycorrhizal fungi (AMF) isolates (Glomus aggregatum, G. fasciculatum, G. intraradices, G. manihotis, G. mosseae, and G. verriculosum) were investigated on seedlings of Acacia senegal (L.) Willd., a multipurpose tree legume highly valued for arabic gum production. Mycorrhizal root colonization, plant growth and relative mycorrhizal dependency (RMD) were measured in A. senegal seedlings growing in soils from three geographical sites in Senegal (Dahra, Bambey and Goudiry) and two soil conditions (sterilized vs unsterilized) in the glasshouse. The impact of inoculation on mycorrhizal root colonization and plant growth depended on AMF isolates, soil origins and soil conditions. Mycorrhizal root colonization and plant growth were increased in sterilized soils regardless of soil origin and AMF isolates. The degree of RMD of A. senegal seedlings varied with soil origin, soil condition and AMF isolates. A. senegal showed the highest RMD values, reaching a maximum of 45 %, when inoculated with G. manihotis. However, in unsterilized soils, no significant effect of AMF inoculation on plant growth was observed despite significant root colonization with certain AMF isolates in Dahra and Goudiry soils. This indicates that the most infective AMF isolates were not the most effective and unsterilized soils may contain effective mycorrhizal propagules. In conclusion, it is important to consider the native mycorrhizal component of the soils before harnessing mycorrhizal inoculation programs for sustainable agroforestry systems.     

Influence of Microbial Inoculants on Seedling Production in Teak (Tectona grandis L.f.)

Abstract

An experiment was conducted in forest college campus, Mettupalayam, India with an objective of studying the influence of microbial inoculants on seedling growth of teak. The fruits were collected from the seed production area in top slip (Tamil Nadu) from 63-year mother trees. The microbial inoculants viz., Azospirillum, Azotobacter, phosphobacteria, AMF, Azospirillum + AMF, Azotobacter+ AMF, phosphobacteria + AMF and an uninoculated control formed the treatments. The inoculants were mixed sand and red soil at 1:1:3 ratio, respectively, for preparing nursery mixture for filling the poly pots. The seeds were sown into the poly pots @ two seeds per pot. The experiment was set up in a randomized Block Design with three replications. Biometric observations on various growth attributes were recorded six months after treatment and also their survival in the main field-tested for one year. Azotobacter gave best performance with respect to shoot length, shoot weight and leaf area and combined inoculation of phosphobacteria and AMF gave higher root length and biomass, dry matter production and fine root weight. The survival was better in seedlings that were inoculated with phosphobacteria and AMF. However, below ground biomass and out planting survival during early phase are critical in any tree species. Hence it is concluded that inoculation of phosphobacteria and AMF conjointly be done for better survival and growth of teak seedlings.     

A low-cost method of mycorrhizal inoculation improves growth of Acacia tortilis seedlings in the nursery

A pot experiment in Kenya examined the effects of mycorrhizal and rhizobial inoculation on the growth and symbiont infection of Acacia tortilis seedlings in unsterile soil, in comparison with sterile and unsterile soil controls. Three mixed, arbuscular mycorrhizal inocula, originating from A. tortilis, Terminalia brownii, T. spinosa and Prosopis juliflora (produced under nursery conditions), were used to inoculate the seedlings, which also received a standard, mixed, rhizobial inoculum. Mycorrhizal infection and plant growth were significantly affected by experimental treatment (p<0.001). Some mycorrhizal infection (presumably with indigenous soil-borne inoculum) occurred in the unsterile soil control, but not in the sterile soil control, indicating that transfer of inoculum between pots in the experiment did not occur. Mycorrhizal formation was significantly more extensive in inoculated plants, which also had better shoot and root growth. 24 weeks after inoculation, plants which had received inoculum of A. tortilis origin were 105 mm tall and their roots were 56% mycorrhizal, whereas the controls most representative of normal nursery treatment were only 77 mm tall and 29% mycorrhizal. Nodulation was erratic and poor in all treatments. The results suggest that mycorrhizal inoculation can improve both mycorrhizal infection and growth of tree seedlings in unsterile nursery soil. The methods of culture and inoculation described here could easily be adopted by nurseries at little cost.     

Improving drought tolerance of <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> seedlings by arbuscular mycorrhizas under glasshouse conditions

Effects of mycorrhizal association on improving tolerance of host plant under stress environments have received attentions in recent years. In this paper, six isolates of AMF (arbuscular mycorrhizal fungus) were inoculated to Casuarina equisetifolia seedlings under glasshouse conditions to investigate the effects of AMF on growth and drought tolerance of host plants. All the six isolates which belong to Glomus showed high mycorrhizal colonization (88.5–96.0%) with C. equisetifolia seedlings. Seedlings were subjected to drought stress without watering for 7 days and survival of the seedlings inoculated with Glomus caledonium Gc90068, G. versiforme Gv9004 and G. caledonium Gc90036 increased by 36.6, 23.3 and 16.6%, respectively compared with uninoculated seedlings. Limited influence of AMF on seedling height growth was found, but the effects of AMF on total biomass increment were very significant; the increment ranged from 25.7 to 118.9% compared with uninoculated treatment, and it was noted that AMF exerted more influences on root biomass than shoot biomass. Based on the changes in physiological and biochemical parameters among different treatments caused by drought stress, it was concluded that AM (arbuscular mycorrhizal) associations improve the drought tolerance of C. equisetifolia seedlings by means of some physiological and biochemical responses, such as lowering permeability of plasma membrane and MDA (malondialdehyde) contents, enhancing concentrations of P nutrition, soluble sugar, soluble protein and activities of POX (Peroxidase) of C. equisetifolia seedlings.     

Inoculation of Acacia holosericea with ectomycorrhizal fungi in a semiarid site in Senegal: growth response and influences on the mycorrhizal soil infectivity after 2 years plantation

The main objectives of this study were to determine the influence of several strains of ectomycorrhizal fungi on the growth of Acacia holosericea in a dry tropical environment and their short-term effect on indicators of ecosystem soil conditions such as biological soil properties (microbial biomass, mycorrhizal soil infectivity). Three fungal strains have been tested: Pisolithus albus IR100, P. albus COI024 and Scleroderma dictyosporum IR109. Ectomycorrhizal inoculation has significantly enhanced the growth (shoot and root biomass) of A. holosericea seedlings after 4 months culture in glasshouse conditions in a disinfected soil. In field conditions, the transplanting shock was less important for the ectomycorrhized trees. After 2 years, the inoculated trees had a better growth than that recorded in the control treatment. Ectomycorrhizal inoculation has significantly stimulated height, leave and wood biomass, root biomass of A. holosericea trees. Moreover, this fungal inoculation has significantly modified the leaf nutrient contents for P, N and phenols. Microbial biomass and mycorrhizal soil infectivity were also larger in the inoculated plots.Ectomycorrhizal inoculation could be of great relevance to improve the reafforestation process of degraded areas with legumes fast growing trees. This biological practice could also ameliorate soil characteristics such as microbial biomass or mycorrhizal soil infectivity.     

Arbuscular mycorrhizal association of indigenous agroforestry tree species and their infective potential with maize in the rift valley, Ethiopia

Tree species in agroforestry are important source of inoculum for companion agricultural crops. Agroforestry trees can serve as a source of Arbuscular mycorrhiza (AM) inoculants to intercropped annuals. We studied spore abundance, root colonization of Albizia gummifera (J.F. Gmel.) and Croton macrostachyus (Hochst Ex Del.) trees and their effect on colonization of maize. Soil and root samples were collected from field standing trees from under and outside the canopy of trees and maize crops in the main rainy season. The number of spore count was significantly higher under the canopy of A. gummifera (791/100 g of dry soil) and C. macrostachyus (877/100 g of dry soil) trees than outside the canopy (547 and 588/100 g of dry soil, respectively). The level of root colonization of C. macrostachyus (45 %) was higher than A. gummifera (41 %). Root colonization of maize crops grown under the canopy of A. gummifera and C. macrostachyus trees was significantly higher than outside the canopy (P < 0.001). Maize seedlings grown on non-sterilized soils collected under and outside the canopy of A. gummifera and C. macrostachyus trees recorded higher root colonization, plant height, shoot and root dry weight than grown on sterilized soils (P < 0.001). The percentage of AM colonized roots of Zea mays seedlings was significantly positively correlated with the number of spore counts for field soils. The rhizospheres of indigenous agroforestry perennial species are important source of inoculum for annuals. The integration of perennials and annuals in an agroforestry system enhances the maintenance of soil quality in the tropics.     

工业和住宅污泥对大叶相思种子萌发以及幼苗生长参数的影响

在孟加拉国吉大港大学林业与环境科学院苗圃内,评价了污泥对大叶相思种子萌发以及幼苗生长的作用。播种前,将污泥与营养匮乏的自然林土壤按不同比率混合,分别在播种后的第1,2和3个月时,记录种子萌发情况和幼苗生长参数（枝条和根系的长度、直径、枝条的鲜重和干重、根和总的干物质量）。在播种前和幼苗采收后,分别测定各个处理的污泥-土壤混合物的理化参数（pH值、有机碳、氮、磷、钾）,以及重金属含量（铬、镍、锰、镉、锌）。结果表明,与对照相比,污泥-森林土壤混合物中播种的种子萌发率和幼苗生长参数发生显著变化。土壤-污泥按2：1混合时,种子萌发率最高（90%）,幼苗生长速率和生物量最大,有机碳和氮磷钾含量也最高。混有工业污泥的土壤中重金属含量要高于混有住宅污泥的土壤。在工业污泥土壤中,污泥-土壤按1：1混合时,重金属含量最高。据此推荐,在退化森林土壤中,土壤-污泥以2：1混合较适合大叶相思种子萌发和幼苗生长。     

丛枝菌根真菌对在含砷土壤中的云南石梓(Gmelina arborea)生长的影响(英文)

孟加拉东南部土壤中的砷含量很高,不仅威胁人的健康,而且对土壤也有破坏作用。云南石梓(Gmelina arborea)在孟加拉是个快速生长的树种,也是含砷土壤中的潜力树种。研究评价了含砷试验土中丛枝菌根真菌对云南石梓(Gmelina arborea)生长的影响。播种前,四种不同浓度的砷(10mg·kg-1、25mg·kg-1、50mg·kg-1和100mg·kg-1)被加入到试验土中。记录生长参数,如,植物的根、苗鲜重、干重、冠幅径、根长和苗高、根瘤菌和孢子菌群落。菌根植株较非菌根植株生长好。与其它含砷量高的土壤中植株的生长情况相比,在含砷量为10-mg·kg-1的土壤中,菌根植株和菌根生长效果最佳,菌根植株生物量最高。随着砷浓度的增加,种苗生长,根瘤菌和孢子菌群落均明显降低p0.05)。与非菌根植株比较,菌根植株高生长增加了40%,生物量增加了2.4倍。研究表明,根瘤菌接种可以减少有害土壤中的云南石梓(Gmelina arborea)的初生长的影响。     

Polylepis australis’ regeneration niche in relation to seed dispersal,site characteristics and livestock density

Establishment of Polylepis forests endemic to the mountains of South America may be affected by seed dispersal, site characteristics and livestock density. Polylepis australis (“tabaquillo”) grows in the high mountains of central Argentina, where we set up 76 square study plots of 900 m². To determine dispersal distance, we distributed 6 seed traps per plot in and around 20 plots. To determine the best site characteristics and livestock stoking rates, at two river basins differing in historic stocking rates, we analyzed the presence of seedlings in 56 plots and recorded topography, vegetation types and indicators of livestock activity. We also measured microsite characteristics in a sample of 32 comparable pairs of 1 m² quadrats, with and without seedlings. Maximum recorded dispersal distance of P. australis seeds was 6 m, and seedlings were found no more than 10 m from seed trees. The numbers of seedlings and seed trees were 3.5 and 4 times higher, respectively, in the basin with less livestock. At the 900 m² plot scale, a Poisson regression indicated a positive relationship between seedling number and P. australis canopy cover. At the quadrat scale (1 m²), seedlings were found in quadrats with significantly lower evidence of soil erosion than comparison quadrats without seedlings. We conclude that the main limitations to recruitment are short seed dispersal distances, lack of seed trees and extreme soil erosion. Management should therefore aim at preserving seed trees and reducing livestock density to prevent erosion.     

Selection of efficient VA mycorrhizal fungi for Casuarina equisetifolia—second screening

An investigation was carried out to screen and select efficient vesicular arbuscular mycorrhizal (VAM) fungi for inoculating the forest tree species, Casuarina equisetifolia. The seedlings were inoculated with 10 different VAM fungi, obtained from various sources. Inoculated seedlings generally had greater plant height, stem girth, biomass and P content than uninoculated plants. They also had more mycorrhizal root colonization and spore numbers in root zone soil. C. equisetifolia seedlings responded best (in biomass) to inoculation with Glomus mosseae (Nicolson and Gerdemann) Gerdemann and Trappe, closely followed by Acaulospora laevis Gerdemann and Trappe and G. fasciculatum (Thaxter Sensu Gerdemann) Gerdemann and Trappe; all the three being statistically on par with each other.     

Do pine plantations provide mycorrhizal inocula for seedlings establishment in grasslands from Patagonia,Argentina?

We investigated if Pinus ponderosa plantations in Patagonia are able to produce viable mycorrhizal inocula towards adjacent grasslands, which only harbor endomycorrhizal vegetation. We hypothesized that these inocula have the potential to contribute to the establishment of naturally disseminated seedlings. Also, we determined the main fungal taxa involved in this process. Seven plantations in the onset of their reproductive phase and located in the Patagonian native forest/steppe ecotone (Argentina) were selected. Soil samplings were obtained at nine points along a 450 m long, W-E transect established in each plantation. Soil bioassays were performed in a greenhouse, with P. ponderosa seedlings acting as hosts for mycorrhizal inocula present in soil samples, during 12 months. Mycorrhization percentage, morphotype richness and morphotype composition was determined through morphological evaluation. Viable ecto- and ectendomycorrhizal inocula were found disseminated outside plantations. The amount of mycorrhizal inoculum followed a decreasing function with distance to plantation edges. Mycorrhizal fungal genus Rhizopogon and “E-strain” mycorrhizal types appeared as pioneering taxa regarding seedlings colonization, being the most persistent and frequent symbionts found. Plantations, thus, facilitate the surrounding terrain for newcoming seedlings through the dispersion of mycorrhizal fungal inocula.     

Biodiversity of arbuscular mycorrhizal fungi in <Emphasis Type="Italic">Amygdalus scoparia</Emphasis> Spach plantations and a natural stand

The biodiversity of arbuscular mycorrhizal fungi (AMF) was surveyed in the Kolm region of Iran in three adjacent sites, a natural stand, a 10-year-old and a 15-year-old plantation of Amygdalus scoparia. To date, there have been few studies of AMF biodiversity in Iran, especially in the western forests of the country. For this study, soil and root samples were taken from A. scoparia rhizosphere soil in spring and autumn. Almost half of the root length was colonized by AMF. We identified 13 AMF species belonging to Glomeraceae, Claroideoglomeraceae or Diversisporaceae. The three plantations differed in terms of soil electrical conductivity, organic C and P. Spore density was significant correlated with P concentration. Root length colonization was correlated only with soil Ca. Species diversity and richness were significantly correlated with soil N, P, organic C and spore density. AMF diversity in 15-year-old plantations was more similar to that in the natural stand than in the 10-year-old plantation. We confirmed that a 15-year-old plantation is not similar in terms of AMF colonization to natural stands. We conclude that more than 15 years are required for AMF colonization of plantations to resemble that of natural stands.     

香格里拉亚高山两种植被类型主要植物的丛枝菌根研究

以滇西北香格里拉亚高山地区生态严重退化的荒草坡植被类型和基本实现恢复的近原生林地植被类型中主要植物的丛枝菌根真菌(AMF)作为研究对象,对这些植物根系的AMF感染率及其根际土壤中的AMF孢子密度进行了调查研究。结果表明,荒草坡的10种植物和近原生林地的9种植物,均形成典型的丛枝菌根(AM)。荒草坡和近原生林植物根际土壤中的平均孢子密度分别为674±221(SE)个/100g土和290±72个/100g土。单因素方差分析表明,两植被类型的主要植物在根系AMF菌丝感染率以及根际土壤中AMF孢子密度方面的差异都极显著。同种植物在近原生林地具有较高的AMF感染率,而根际土壤中的孢子密度则是在荒草坡为高。相关性分析表明,所调查植物的根系AMF菌丝感染率与根际土壤中的AMF孢子密度间不存在相关性。     

Effects of arbuscular mycorrhizal fungi on the drought tolerance of Cyclobalanopsis glauca seedlings under greenhouse conditions

Cyclobalanopsis glauca is an important afforestation tree species that is widely used for revegetating the karst region of southwest China. Vegetation in this region is regularly commonly subjected to drought stress because of the geology and water shortages. Here, we investigated the influence of two arbuscular mycorrhizal fungi (AMF) Glomus mosseae and Glomus intraradices on the drought tolerance of C. glauca seedlings under greenhouse conditions. AMF-treated and non-AMF-treated C. glauca seedlings were maintained under two different water regimes (well watered: 80 % field capacity; drought stress: 40 % field capacity) for 90 days. The AMF colonization rate was higher under well-watered conditions compared to drought stress conditions. The growth and physiological performance of C. glauca seedlings were significantly affected by drought stress. Under drought stress conditions, mycorrhizal seedlings had greater height, base diameter, leaf area, and biomass compared to non-mycorrhizal seedlings. In addition, under drought conditions, AMF-inoculated seedlings had greater superoxide dismutase and peroxidase activity, higher soluble sugar content, and lower proline content compared to non-inoculated seedlings. Furthermore, AMF colonization increased the phosphorus and potassium content of seedling shoots under both well-watered and drought stress conditions. Therefore, AMF colonization enhanced the drought tolerance of C. glauca seedlings by improving growth performance, nutrient content, the quantity of osmotic adjustment compounds, and antioxidant enzyme activity. The results indicate that AMF are of potential use for the restoration of vegetation in the karst region of southwest China.     

Occurrence of dark septate endophytes in Nothofagus seedlings from Patagonia,Argentina

Temperate forests of southern Argentina and Chile cover a wide latitudinal and altitudinal range on both sides of the Andes, with Nothofagus species being one of the main tree species. Most of the larger remnants are protected within the limits of national parks and forest reserves. However, some of these are invaded by exotic conifers such as Pseudotsuga menziesii. In order to better understand the mycorrhizal condition within the Nothofagus–P. menziesii interaction, transects were established at six study sites along the effective recruitment area. Three treatments were established: Nothofagus forest without P. menziesii invasion (Forest), Nothofagus–P. menziesii matrix (Interface) and P. menziesii plantation lacking Nothofagus specimens (Plantation). A 2 kg mixed soil sample was taken at each treatment site and kept in brand-new plastic bags. A soil bioassay with Nothofagus seedlings acting as baits was set to evaluate the mycorrhizal inoculum potential and soil-fungi composition at each sampling treatment. Forty percent of the evaluated seedlings were found to be colonised by dark septate endophytes (DSE) fungi. Ectomycorrhizal colonisation percentage was significantly higher in plants growing in Forest soils (mean = 57.77%) when compared with Interface (mean = 42.53%) and Plantation (mean = 44.65%). The high incidence of DSE in young roots of Nothofagus seedlings in this study supports the hypothesis that DSE might be pioneering colonisers of young tree seedlings in secondary successional environments. For future interventions in the forest, either with productive or protective intentions, it should be taken into account the incidence of these fungi, which may have an important positive role in Nothofagus forest post-invasion recovery.     

Influence of the fertilisation method in controlled ectomycorrhizal inoculation of two Mediterranean pines

The influence of the fertilisation method: soluble (SF) vs. slow-release fertiliser (SRF) and of inoculation with Laccaria laccata (Scop.) Fr., Pisolithus tinctorius (Pers.) Coker & Couch and Melanogaster ambiguus (Vittad.) Tul & C. Tul. on ectomycorrhizal colonization and growth of Pinus pinea L. and Pinus pinaster Ait. was evaluated. For both pines, mycorrhization with L. laccata was not affected by the fertilisation method. Percentages of ectomycorrhizas (ECM) formed by P. tinctorius were dependent on the fertilisation method, the inoculum type (vegetative or spores) and the pine species involved. ECM formed by M. ambiguus were increased with fertilisation in both pines. Inoculation significantly improved P. pinea biomass when seedlings were fertilised with SRF whereas no effect was found in non-fertilised ones. For non-fertilised P. pinaster, inoculation with L. laccata and both inocula of P. tinctorius increased seedling biomass whereas fertilisation neutralised the fungal effect. Fertilisation increased P. pinea and P. pinaster biomass, independently of the inoculation treatment.