Greenhouse production of Burgundy truffle mycorrhizae on oak roots

Truffles, subterranean fruit-bodies of the ectomycorrhizal genus Tuber, are currently cultivated on host trees in plantations, both to offset declining wild production and to extend their geographic distribution. We report the effects of the potting medium and potting method components of two very different seedling production systems on the colonization of Quercus bicolor × Q. robur seedlings by T. aestivum (the Burgundy truffle). Seedlings grown in book planter containers using a peat-based medium were smaller yet much better colonized by T. aestivum than seedlings produced in RPM^® (Root Production Method) containers using a ground bark-based RPM medium. In either container method, the peat-based medium supported development of more T. aestivum mycorrhizae than did the RPM medium. Seedlings grown in book planters developed significantly more root tips l^?1 of medium than did seedlings grown in RPM containers. The optimum pH for T. aestivum mycorrhiza development in the peat-based medium is between 6.7 and 7.5.     

Seed source and vesicular-arbuscular mycorrhizal symbiont affects growth of Juglans nigra seedlings

Seedlings of Juglans nigra from three seed sources were grown in fumigated soil without vesicular-arbuscular mycorrhizal (VAM) fungi or inoculated with Gigaspora margarita, Glomus deserticola or Glomus etunicatum. Vesicular-arbuscular mycorrhizal development varied significantly between fungal symbionts within a black walnut source. Glomus deserticola and G. etunicatum produced the highest levels of root colonization in all sources. Significant differences in seedling shoot and root growth were attributed to root colonization by specific VAM fungi in each black walnut source. Glomus deserticola stimulated seedling leaf area and root weight 26 and 52%, respectively, in one seed source. Seedling leaf N, P and K concentrations were significantly improved by VAM in two seed sources. Juglans nigra seedlings respond favorably to VAM colonization. However, differences between seed sources suggest a strong host-symbiont interaction.     

Field survival of containerized red and jack pine seedlings inoculated with mycelial slurries of ectomycorrhizal fungi

Red pine and jack pine seedlings growing in styroblocks were inoculated 8 wk after sowing with mycelium/agar slurries of 3 mycorrhizal fungi (Laccaria bicolor, Scleroderma citrinum, and an unidentified basidiomycete), and one suspected mycorrhizal fungus (Cantharellula umbonata). Seedlings inoculated with L. bicolor developed mycorrhizae earlier and in greater numbers than the other inoculation treatments, with red pine out-performing jack pine in both respects. At 34 wk following sowing, seedlings were outplanted on a cleared xeric site in Baraga Co., in Michigan's Upper Peninsula. Seedlings inoculated with C. umbonata failed to form mycorrhizae and were not outplanted. Inoculation treatments did not affect shoot or root weight at outplanting. Red pine inoculated with L. bicolor averaged 21% and 19% greater survival compared with control seedlings after one and two years in the field, respectively. Other inoculation treatments failed to increase seedling survival for either tree species. Jack pine demonstrated higher overall survival than did red pine for both years in all corresponding inoculation treatments.     

Growth, nitrogen fixation and mineral acquisition of Alnus sieboldiana after inoculation of Frankia together with Gigaspora margarita and Pseudomonas putida

The role of tetrapartite associations among Frankia, Gigaspora margarita (an arbuscular mycorrhizal fungus), Pseudomonas putida (rhizobacterium), and Alnus sieboldiana in growth, nitrogen fixation, and mineral acquisition of A. sieboldiana was investigated. Seedlings of A. sieboldiana were inoculated with Frankia isolated from root nodules of alder, followed by inoculation of G. margarita and P. putida, and were grown for 5 months in a greenhouse. The seedlings inoculated with Frankia and G. margarita together produced the highest biomass of shoots and root nodules. Nitrogen-fixation activity, measured by acetylene reduction assay, was observed when Frankia was inoculated. The activity, on a per-nodule gram basis, decreased after G. margarita inoculation, but on a per-plant basis there was no significant difference in the activity among inoculation treatments. The mineral content in the seedlings changed after inoculation with Frankia, but not after inoculation with P. putida and/or G. margarita. The results showed a synergistic interaction among Frankia, the mycorrhizal fungus, and the rhizobacterium on the growth of A. sieboldiana.     

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.     

Root diseases in bareroot and container nurseries of the Pacific Northwest: epidemiology, management, and effects on outplanting performance

In forest and conservation nurseries in the Pacific Northwest USA, seedling production can be limited by root diseases caused by fungi in the genera Fusarium Link:Fr., Cylindrocarpon Wollenw., Phytophthora de Barry, and Pythium Pringsh. Fusarium, Cylindrocarpon, and Pythium are the most ubiquitous, whereas incidence of Phytophthora is mostly associated with coastal bareroot nurseries. All of these root pathogens are encouraged by water saturated soils or media. Seedlings infected with Fusarium, Phytophthora, or Pythium often appear chlorotic or necrotic with extensive root decay. Cylindrocarpon often causes serious root decay without shoot symptoms. The best approach to mitigate losses from these diseases is to use a holistic integrated pest management program. This program should combine chemical controls with cultural practices, particularly those that increase soil permeability and drainage and reduce potential sources of inoculum, especially by disinfesting seeds and containers reused for crops. In general, we found that seedlings meeting nursery specifications for outplanting on forest soil (proper height, root collar diameter, healthy shoot color, lack of disease symptoms) but having these disease organisms on their root systems perform as well as non-infected seedlings.     

Effect of Frankia inoculation on the growth of Alnus sieboldiana on unsterilized soil

The effect of inoculation with Frankia, a N-fixing actinomycete, on the growth of Alnus sieboldiana seedlings was studied on unsterilized soil from a nursery and an alder stand (forest of Alnus firma). The seedlings of A. sieboldiana were inoculated with Frankia before or after a 2-month culture on sterilized vermiculite, during which they nodulated, and transplanted to unsterilized soil from the nursery and the alder stand. The control seedlings were also cultured on sterilized vermiculite for about 2 months and transplanted to unsterilized soil without Frankia inoculation. The seedling growth, nodulation and N-fixing activity were measured 3, 10 and 16 weeks after the transplantation. Growth and nodule biomass of the seedlings inoculated with Frankia and those grown on the alder soil were better than those without inoculation with Frankia and grown on the nursery soil, respectively. The seedlings inoculated before spontaneous nodulation grew better than those inoculated at the transplantation. Nitrogen-fixing activity measured by acetylene reduction assay at 16 weeks after the transplantation was higher in the seedlings grown on the soil from the nursery than on the soil from the alder stand.     

Influence of water-stress acclimation and <Emphasis Type="Italic">Tuber melanosporum</Emphasis> mycorrhization on <Emphasis Type="Italic">Quercus ilex</Emphasis> seedlings

Mycorrhizal and non-mycorrhizal holm oak (Quercus ilex L.) seedlings inoculated with black truffle (Tuber melanosporum) were grown under nursery conditions and subjected to drought hardening for 4 months in autumn and winter followed by irrigation for 10 days. Leaf water potential and stomatal conductance were monitored during the 4 months of drought. When the test was completed (March), measurements were made for each treatment (inoculated or non-inoculated), and watering regime (watered and water-stressed). Pressure–volume curves, osmotic potential at full turgor, osmotic potential at zero turgor and the tissue modulus of elasticity near full turgor were calculated. Mycorrhizal colonization and growth, and the content of the main mineral nutrients N, P, K, Ca and Mg were measured. Water stress affected plant growth, caused an elastic adjustment of the plant tissues, and decreased the P and K content, and inoculation improved the nitrogen content. Drought acclimation apparently achieved the goal of improving the drought tolerance of holm oak seedlings, without depressing ectomycorrhizal root colonization by T. melanosporum. José Antonio Rodríguez Barreal—Deceased     

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.     

Improvement of early growth of two tropical peat-swamp forest tree species Ploiarium alternifolium and Calophyllum hosei by two arbuscular mycorrhizal fungi under greenhouse conditions

Tropical peat-swamp forests are one of the largest near-surface reserves of terrestrial carbon. However, many peat-swamp forest tree species have resulted in the reduction due to over-exploitation, forest fires and conversion into agricultural land in Indonesia. The objective of this study was to determine the effects of two arbuscular mycorrhizal (AM) fungi, Glomus clarum and G. aggregatum, on the early growth of two slow-growing peat-swamp forest tree species, Ploiarium alternifolium and Calophyllum hosei, under greenhouse conditions. Cuttings of P. alternifolium and C. hosei were uninoculated or inoculated with G. clarum and G. aggregatum and grown under greenhouse conditions for 6 months. Percentage AM colonization, plant growth, phosphorus (P) concentration and survival rate were measured. The AM colonization of P. alternifolium and C. hosei ranged from 27 to 32% and 18 to 19%, respectively. Colonization by G. clarum and G. aggregatum increased shoot height, stem diameter, leaf number, and shoot and root dry weights. Cutting shoot P content were increased by AM fungal colonization. The survival rates of inoculated plants were higher (100%) than those of control plants (67%). The results suggest that inoculation with AM fungi improves early growth of P. alternifolium and C. hosei in a tropical peat-swamp forest and can therefore contribute to rehabilitation of peat-swamps.     

Effect of the inoculation and distribution of mycorrhizae in <Emphasis Type="Italic">Plathymenia reticulata</Emphasis> Benth under monoculture and mixed plantation in Brazil

This paper investigates the distribution of arbuscular mycorrhizal fungi (AMF) spores and AMF colonization in a field study in southeastern Brazil. Response to AMF and rhizobial inoculation was studied in monocultures of Plathymenia reticulata and mixed plantations with both Tabebuia heptaphylla and Eucalyptus camaldulensis in a sandy soil during two consecutive years. P. reticulata height and diameter and mycorrhizal colonization and AMF diversity were measured in dry and rainy periods. The inoculated treatment of E. camaldulensis, T. heptaphylla and P. reticulata mixed plants showed higher height and diameter growth of P. reticulata used as well as increased root colonization and AMF spore numbers. Spore populations were found to belong to five genera: Acaulospora, Entrophospora, Glomus, Gigaspora and Scutellospora, with Glomus dominating. Agroforestry practices including use of leguminous tree P. reticulata effectively maintained AMF spore numbers in soils and high AMF colonization levels compared with monocultures, proving an efficient system for productivity and sustainability.     

Soil factors influencing the growth response of <Emphasis Type="Italic">Acacia holosericea</Emphasis> A. Cunn. ex G. Don to ectomycorrhizal inoculation

Australian acacias like Acacia holosericea are excellent candidates for the revegetation of arid zones in Africa. Their high ability to develop multiple symbioses with soil microorganisms is crucial to their rapid development in adverse climatic and edaphic conditions. These symbioses include nitrogen fixation with rhizobia, vesicular arbuscular mycorrhization and ectomycorrhization. We set up an experiment to test the growth response of A. holosericea to ectomycorrhizal inoculation in 13 different soils from Senegal. After autoclaving the soil, the experiment was conducted in a greenhouse for four weeks. Plants were inoculated with Pisolithus albus strain IR100. The following parameters were measured: plant biomass, N, P, K and Ca foliar composition, spontaneous nodulation rate, and ectomycorrhizal colonization. Data were analyzed in light of the physical, chemical and total microbial characteristics of the soil. The results demonstrated a global promoting effect of P. albus inoculation on plant (shoot and root) growth and on foliar P and K, together with a depressive effect on N, while Ca rates were barely affected. Interestingly, spontaneous nodulation with putative water- or airborne rhizobia was stimulated after P. albus inoculation. However, these nodules seemed poorly effective, as they failed to cause any change in plant growth or in foliar N composition within the nodulated and nonnodulated control plants. These results show that plant growth response to ectomycorrhizal inoculation is greatly dependent on soil characteristics, and that root ectomycorrhizal colonization is influenced by biotic factors such as soil microbiota. From a practical point of view, data from the present study demonstrate that it is possible to optimize the effect of fungi on plant growth by screening soils under nursery conditions.     

Growth and nutrient concentration of two native forage legumes inoculated with Rhizobium and Mycorrhiza in Missouri, USA

The Center for Agroforestry at the University of Missouri has tested numerous native legumes for potential use in agroforestry and selected Illinois bundleflower (Desmanthus illinoensis (Michaux) MacMillan ex Robinson and Fern.) and panicled tick clover (Desmodium paniculatum (L.) DC.) for further testing. Our objective was to document the effect of arbuscular mycorrhizae (AM) (Glomus spp.) and Rhizobium on growth and nutrient concentration of these legumes. Seeds were planted in a greenhouse and inoculated with one of two species of AM and/or one of two strains of Rhizobium. Plants were harvested after 80 d and data taken on leaf and stem dry weight, root fresh weight, stem height, nodulation, AM colonization, and N, P, K, Ca, and Mg concentration. Inoculation with Rhizobium did not affect plant growth in Illinois bundleflower, but colonization by Glomus intraradices increased all plant growth variables except stem height. Nutrient concentration was unaffected by the presence of either endophyte. In contrast, inoculation of panicled tick clover with Rhizobium str. 41Z10 increased leaf dry weight (32%) compared to the control and root fresh weight (41%) compared to str. 32Z3, and colonization by G. intraradices increased leaf dry weight (35%) and stem height (26%). Both species of AM increased P and K concentration (41% and 55%, respectively) in panicled tick clover. Our results suggest that the growth of these legumes can be improved by the use of proper AM species and/or Rhizobium strains. However, additional research to identify the best Rhizobium and AM inoculates for these plant species is important in developing strategies for their use in agroforestry. This revised version was published online in June 2006 with corrections to the Cover Date.     

Biosolids,mycorrhizal fungi and eucalypts for phytostabilization of arsenical sulphidic mine tailings

The effect of the addition of biosolids combined with a native arbuscular mycorrhizal inoculum on growth of a eucalypt (Eucalyptus cladocalyx), and on trace element stabilization of arsenical sulphidic gold mine tailings was tested. A glasshouse trial was established using four substrates: tailings (T); tailings with a layer of topsoil (TS); tailings amended with 100 t ha⁻¹ biosolids (LB), and tailings amended with 500 t ha⁻¹ biosolids (HB). Half of the pots were inoculated with a mixture of Glomus sp. (WUM51–9227), Scutelospora aurigloba (WUM51–53) and Acaulospora laevis (WUM46) culture mix, and others were uninoculated controls. Two seeds per pot were sown. Leaf and stem weights, leaf area and plant height were measured for each plant. Nutrients and trace element concentrations in leaves and stems were determined. Addition of biosolids significantly increased mycorrhizal colonization (both ectomycorrhizal and arbuscular mycorrhizal) in roots. Biosolids clearly increased the very early establishment and growth of the eucalypts. In general, mycorrhizal inoculum increased plant biomass production and nutrient uptake. Trace element concentrations in trees of the biosolid-amended pots were in general high, however, mycorrhizal inoculation could reduce the uptake of toxic elements by plants.     

Early root morphology of jack pine seedlings grown in different types of container

Abstract

This study compared the effects of container type on early root system morphology of jack pine (Pinus banksiana Lamb.) seedlings to determine impacts of container type on root characteristics that may be important for tree stability. Seedlings were grown for one season in Multipots®, Ventblocks®, Copperblocks®, Starpots® and Jiffy® pellets, and for a second season in sand culture in pots. After the first growing season, roots that had grown between Jiffy plugs were cut using a knife, either “early”, in September, or “late”, in November. Seedlings produced in non-pruning containers, Multipots and Ventblocks, had more vertical structural roots, less even root distribution and high shoot:root mass after a second season. Seedlings grown in pruning containers, Copperblocks, Starpots and Jiffy pellets, had more horizontal structural roots and more evenly distributed roots. Jiffy seedlings had high shoot:root mass after pruning, but ratios decreased to low levels similar to Starpots and Copperblocks after a second season in pots. Early pruning of Jiffy seedlings removed less root mass than late pruning, but total live root mass of early and late pruned seedlings was similar after the potting trial. Pruning containers thus produced structural root forms with more desirable characteristics for tree stability. These characteristics were evident after the first season and persisted after the second season in the potting trial.     

蓝桉和尾叶桉混合菌根研究 Ⅱ.混合菌根的接种效应

接种苗木在生长量和生物量上均表现出极其显著的生长优势,ＥＣＭ真菌蜡蘑菌单接种及其与４种ＶＡ菌根菌剂混合接种,对苗木生长的促进作用尤为显著。在接种后１６周时,与对照苗相比,蓝桉和尾叶桉接种苗木的高生长量最大增幅分别为２８．８６％（ＬＳ）和８６．６５％（ＬＧ）;两种桉树地上部分平均干质量最大增幅分别为１２９．９３％（ＬＳ）和１３３．３４％（Ｌ）,地下部分分别为１１９．９３％（ＬＦ）和１７４．８３％（Ｌ     

Production of <Emphasis Type="Italic">Pinus halepensis</Emphasis> seedlings inoculated with the edible fungus <Emphasis Type="Italic">Lactarius deliciosus</Emphasis> under nursery conditions

Inoculation with edible fungi bestows an added value on mycorrhized plants since production of mushrooms as a secondary crop can be an economically-valuable resource in forestry management. In order to establish a protocol for controlled mycorrhization of Pinus halepensis with the edible fungus Lactarius deliciosus, several factors such as fertilisation (35, 60 or 120 mg total N and 13.5, 27 or 54 mg total P), potting substrate (sphagnum peat or sphagnum peat/vermiculite) and fungal inoculum (mycelial slurry produced in a bioreactor, alginate beads or peat/vermiculite) were assessed. The most effective inoculum was mycelial slurry at a dose of 10 ml/plant. The two potting substrates assessed were compatible with mycorrhiza formation. The greatest number of mycorrhized seedlings was obtained with a moderate level of N (35 mg/plant) or P fertilisation (27 mg/plant). Inoculation did not produce a consistent growth effect on P. halepensis, but seedlings met the quality requirements, making them suitable for planting out.     

Growth response and nutrient utilization of <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> seedlings inoculated with bioinoculants under tropical nursery conditions

We investigated the role of tetrapartite associations between an arbuscular mycorrhizal (AM) fungus (Glomus geosporum), phosphate solubilizing bacteria (Paenibacillus polymyxa), Frankia and Casuarina equisetifolia on growth, nutrient acquisition, nutrient utilization and seedling quality of C. equisetifolia. Seedlings of C. equisetifolia were grown in an Alfisol soil and inoculated with G. geosporum, P. polymyxa and Frankia either individually or in combinations. Inoculation of bioinoculants stimulated seedling growth, the efficiency of nutrient uptake and improved seedling quality. However, microbial inoculation generally reduced the efficiency of nutrient utilization in dry matter production (nutrient use efficiency). Inoculation of P. polymyxa or Frankia increased the extent of AM colonization, which resulted in the accumulation of the nutrients. Seedlings inoculated with Frankia and G. geosporum had more, and heavier nodules compared to seedlings inoculated with Frankia alone. Dual inoculation of microbes was more effective than individual inoculations. The growth response of seedlings to inoculation involving all the microbes was greater than the response to either individual or dual inoculations. The results of this study showed that the tetrapartite association could improve the growth, nutrient acquisition and seedling quality of C. equisetifolia under tropical nursery conditions.     

Gliocladium virens in an alginate prill ineffective as a biological control of Fusarium root disease in container-grown Douglas-fir

An alginate prill formulation of Gliocladium virens (GL-21) was added as a top-dressings (54 g per m²) or incorporated into medium (1.2 kg per m³) used to grow Douglas-fir seedlings in styrofoam containers. Seedlings in the top-dress treatment were similar to control seedlings; infection and colonization by naturally-occurring Fusarium was unaffected by treatment. Incorporated G. virens reduced seedling growth and increased occurrence and colonization intensity of Fusarium. In a laboratory experiment, inoculating Douglas-fir seedlings with G. virens (10% w/w) prior to inoculation with Fusarium increased survival time when compared to concurrent inoculations of fungi.University of Idaho, Idaho Forest, Wildlife and Range Experiment Station Contribution No. 742.     

Truffle seedling production method has long-term consequences for tree growth and root colonization

Cultivation of Burgundy black truffles (Tuber aestivum syn. T. uncinatum) in Midwestern agroforestry has the potential to provide important income not only to landowners, but also to the hospitality industry. Economically viable harvest depends upon both successful seedling colonization by the truffle fungus and successful growth of the fungus along with the extending root system. We evaluated an established hybrid oak (Quercus bicolor × Q. robur) truffière 5 and 6 years after planting in May 2005, to evaluate the effects of three seedling production methods on tree growth and root colonization by T. aestivum. Oak seedlings produced using two variations on the RPM^© (Root Production Method) substrate grew significantly faster in height and diameter, but were significantly less well colonized by the truffle fungus compared with saplings from seedlings produced by the “Typical” method. We found that 0.7–19.8 % of root tips (mean 5.1 %) sampled from saplings grown from Typical seedlings were colonized by T. aestivum. By 2010, two distinct canopy forms had developed: ‘upright’ versus ‘spreading’. Although root colonization was not related to canopy form, future fruit body production may be influenced by the greater soil shading provided by the spreading canopy form. A comparison of autumn/winter air temperatures in Missouri, USA with European conditions suggests that fruit body production in Missouri will likely be greatest from mid-September through mid-December. Soil pH modification by application of crushed limestone prior to planting was effective in maintaining average pH at 7.16 through 2010, a level consistent with fruit body production.