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Ectomycorrhizas associated with a relict population of Dryas octopetala in the Burren, western Ireland. I. Distribution of ectomycorrhizas in relation to vegetation and soil characteristics.

The distribution of ectomycorrhizas on Dryas octopetala L in grass heaths of the 450 km2 karst region known as the Burren in Western Ireland was examined in relation to soil factors and vegetation type. Ectomycorrhizas were identified or characterised from 56 soil cores from 30 sites, and the occurrence of each ectomycorrhizal (EM) type was quantified by estimating the total length of mycorrhizal tips of each type. Soil organic matter, total nitrogen, extractable phosphorus, pH and depth were the soil factors determined. In total, 24 EM types were recorded. The EM community of Dryas roots was significantly more species-rich in one vegetation type--Hyperico-Dryadetum--than in others (Arctostaphylo-Dryadetum or Asperulo-Seslerietum). Multiple linear regression analyses indicated that soil organic matter and soil depth explained a significant portion of the variation in EM abundance, while soil organic matter and extractable phosphorus explained a significant portion of the variation in EM diversity. Canonical correspondence analysis showed that some individual EM types (e.g. Craterellus lutescens, Cenococcum geophilum, Tomentella sp., Boletus sp.) exhibited distinct soil preferences, most markedly in relation to soil organic matter, which, in this analysis, was the main significant soil variable distinguishing the three vegetation types.

Ascomycota↗

Mycorrhization helper bacteria: a case of specificity for altering ectomycorrhiza architecture but not ectomycorrhiza formation.

Mycorrhization helper bacteria (MHB), isolated from phylogenetically distinct ectomycorrhizal symbioses involving Lactarius rufus, Laccaria bicolor or Suillus luteus, were tested for fungus specificity to enhance L. rufus-Pinus sylvestris or L. bicolor-P. sylvestris mycorrhiza formation. As MHB isolated from the L. rufus and S. luteus mycorrhiza were originally characterised using a microcosm system, we assessed their ability to enhance mycorrhiza formation in a glasshouse system in order to determine the extent to which MHB are system-specific. Paenibacillus sp. EJP73, an MHB for L. rufus in the microcosm, significantly enhanced L. bicolor mycorrhiza formation in the glasshouse, demonstrating that the MHB effect of this bacterium is neither fungus-specific nor limited to the original experimental system. Although the five MHB strains studied were unable to significantly enhance L. rufus mycorrhiza formation, two of them did have a significant effect on dichotomous short root branching by L. rufus. The effect was specific to Paenibacillus sp. EJP73 and Burkholderia sp. EJP67, the two strains isolated from L. rufus mycorrhiza, and was not associated with auxin production. Altered mycorrhiza architecture rather than absolute number of mycorrhizal roots may be an important previously overlooked parameter for defining MHB effects.

Bacteria↗

Effects of simulated rain acidity on ectomycorrhizae of red spruce seedlings potted in natural soil.

Formation of ectomycorrhizae of red spruce (Picea rubens) grown in natural soil was measured after seedlings were exposed to 25 or 50 applications of simulated rain of pH 5.5, 3.5 or 2.5. Ectomycorrhizae were quantified as the total number of ectomycorrhizal tips per centimeter, and as the number of ectomycorrhizal tips for each morphotype and for Cenococcum geophilum. Rain solutions were applied to the soil alone, to foliage and stem alone, or to entire potted seedlings. Final soil pH was linearly related to rain solution acidity. Lower base saturation, calcium and zinc content, and higher exchangeable acidity were observed after pH 2.5 treatments if the soil was exposed. Rain solutions and the subsequent changes in soil characteristics did not affect the total numbers of ectomycorrhizal tips. Four morphotypes of ectomycorrhizae observed for these seedlings were unaffected by simulated rain. However, the numbers of ectomycorrhizal tips formed by C. geophilum tended to increase with rain solution acidity after 50 applications. Method of rain deposition did not affect ectomycorrhizae, suggesting both plant and soil mediated responses may favor certain mycobionts. The results of this study indicate that short-term acidic deposition does not induce significant changes in the frequency of ectomycorrhizae, but higher numbers of C. geophilum tips suggest there may be changes in the relative occurrence of specific morphotypes of fungus species.

Journal Article↗

Identification of symbiosis-regulated genes in Eucalyptus globulus-Pisolithus tinctorius ectomycorrhiza by differential hybridization of arrayed cDNAs.

Ectomycorrhiza development alters gene expression in the fungal and plant symbionts. The identification of a large number of genes expressed exclusively or predominantly in the symbiosis will contribute greatly to the understanding of the development of the ectomycorrhizal symbiosis. We have constructed a cDNA library of 4-day-old Eucalyptus globulus-Pisolithus tinctorius ectomycorrhiza and sequenced 850 cDNAs cloned randomly or obtained through suppression subtractive hybridization (SSH). Based on the absence of a database match, 43% of the ectomycorrhiza ESTs are coding for novel genes. At the developmental stage analysed (fungal sheath formation), the majority of the identified sequences represented 'housekeeping' proteins, i.e. proteins involved in gene/protein expression, cell-wall proteins, metabolic enzymes, and components of signalling systems. We screened arrayed cDNAs to identify symbiosis-regulated genes by using differential hybridization. Comparisons of signals from free-living partners and symbiotic tissues revealed significant differences in expression levels (differential expression ratio >2.5) for 17% of the genes analysed. No ectomycorrhiza-specific gene was detected. The results successfully demonstrate the use of the cDNA array and SSH systems as general approaches for dissecting symbiosis development, and provide the first global picture of the cellular functions operating in ectomycorrhiza.

Basidiomycota↗

[Effect of ectomycorrhizae on the growth of Picea koraiensis seedlings].

Basidioscarps of Agaricales in different Picea koraiensis forest plantations were collected during August-October, 2000. 36 isolaters of species of Agaricales were obtained by isolating and culturing to the basidioscarps. Through indoor inoculation test on seedlings of Picea koraiensis, 6 ectomycorrhizae fungi cultures were obtained from 36 isolaters. The inoculation results show that the period for ectomycorrhizae inoculation to 1-year seedlings of Picea koraiensis should be about 30 days after seedlings emerging, the suitable temperature for ectomycorrhizae forming is about 20 degrees C. 6 ectomycorrhizae strains all have growth-promoting effect to the seedlings of Picea koraiensis. The contents of chlorophyll a of the seedlings inoculated strains of Agaricus silvaticus, 031 and L15 were significantly higher than other strains and control. The contents of chlorophyll b in the seedlings inoculated strains 009, 004, Agaricus silvaticus and L15 were significantly higher than other strains and control. The weights of seedlings which inoculated strains 009, 025, 031, Agaricus silvaticus and L15 were significantly different to control, the weight of seedlings inoculated strains of Agaricus silvaticus and L15 are 19.23% and 23.08% more than control; The heights of the seedlings inoculated 6 strains all have significant difference to control, the weight of seedlings inoculated strains of Agaricus silvaticus and L15 are 17.83% and 16.37% more than control. The results of outdoor inoculation show that the seedlings inoculated Agaricus silvaticus grow best on height, 9.25% more than control after inoculated 70 days; the seedlings inoculated strain L15 grow best on collar diameter, 9.92% more than control after inoculated 70 days; the lateral root numbers of seedlings inoculated strain 009 is largest, 51.91% more than control after inoculated 70 days; the main roots of seedlings inoculated strain 009 are longest, 3.36% more than control after inoculated 70 days; the ectomycorrhizae fungi infestation percentage of seedlings inoculated strain 031 is highest, 13.90% more than control after inoculated 70 days; the photosynthesis of seedlings inoculated strain 031 is more powerful, 20.02% more than control after inoculated 70 days. To sum up, strain 009, strain 031 and strain L15 should be fine strains to seedlings of Picea koraiensis.

Biomass↗

Ectomycorrhiza-mediated repression of the high-affinity ammonium importer gene AmAMT2 in Amanita muscaria.

A main function of ectomycorrhizas, a symbiosis between certain soil fungi and fine roots of woody plants, is the exchange of plant-derived carbohydrates for fungus-derived nutrients. As it is required in large amounts, nitrogen is of special interest. A gene (AmAMT2) coding for a putative fungal ammonium importer was identified in an EST project of functional Amanita muscaria/poplar ectomycorrhizas. Heterologous expression of the entire AmAMT2 coding region in yeast revealed the corresponding protein to be a high-affinity ammonium importer. In axenically grown Amanita hyphae AmAMT2 expression was strongly repressed by nitrogen, independent of whether the offered nitrogen source was transported by AmAMT2 or not. In functional ectomycorrhizas the AmAMT2 transcript level was further decreased in both hyphal networks (sheath and Hartig net), while extraradical hyphae revealed strong gene expression. Together our data suggest that (1) AmAMT2 expression is regulated by the endogenous nitrogen content of hyphae and (2) fungal hyphae in ectomycorrhizas are well supported with nitrogen even when the extraradical mycelium is nitrogen limited. As a consequence of AmAMT2 repression in mycorrhizas, ammonium can be suggested as a potential nitrogen source delivered by fungal hyphae in symbiosis.

Amanita↗

Importance of the ectomycorrhizal network for seedling survival and ectomycorrhiza formation in rain forests of south Cameroon.

An investigation was undertaken at two sites south Cameroon to assess the importance of living roots of adult trees as sources of inoculum for survival, ectomycorrhizal colonisation and growth of seedlings of Paraberlinia bifoliolata. One-month-old seedlings of Paraberlinia bifoliolata, isolated from or in contact with roots of adult target trees, were transplanted in concentric circles 5, 10, 15, and 30 m away from the stem bases of four adult tree species (Afzelia bipindensis, Brachystegia cynometroides, Paraberlinia bifoliolata and Tetraberlinia bifoliolata). After 4 and 8 months, survival, ectomycorrhiza formation and seedling height were observed; plant biomass was determined after 8 months. After 4 months, there was no difference in seedling survival between seedlings in contact with roots of adult trees and isolated seedlings. The number of ectomycorrhizal seedlings was higher among seedlings in contact with adult trees than among isolated seedlings. After 8 months, both seedling survival and ectomycorrhiza formation were higher in seedlings in contact with roots than in isolated seedlings. Seedling survival and ectomycorrhiza formation were highest under Brachystegia. The fraction of surviving seedlings that had formed ectomycorrhizas was lowest under Afzelia. The fraction of surviving seedlings that were ectomycorrhizal declined monotonically towards the edge of the crown projection. Biomass of seedlings in contact with adult trees was significantly higher than that of isolated seedlings. This difference was consistent for all four tree species. The height of seedlings did not vary with the treatment.

Cameroon↗

Ectomycorrhizae between Alnus acuminata H.B.K. and Naucoria escharoides (Fr.:Fr.) Kummer from Argentina.

Field ectomycorrhizae of Naucoria escharoides on Alnus acuminata ("andean alder", "aliso del cerro") are described in detail for the first time. Naturally occurring ectomycorrhizal roots were sampled beneath sporocarps of N. escharoides. The samples were taken from four natural forest plots at two homogeneous A. acuminata sites (Tucumán and Catamarca Provinces, Argentina). The ectomycorrhizae were characterized morphologically and compared by means of PCR/RFLP analysis of the internal transcribed spacer region of the nuclear rDNA. The most important morphological features of the ectomycorrhizae are a white to pale yellow mantle, simple to monopodial branches, hyaline emanating hyphae, abundant hyphal bundles emerging more or less perpendicularly from a plectenchymatous mantle, and an acute or rounded apex with or without a mantle. N. escharoides fruitbodies have white basal mycelium with emanating hyphae similar to those of andean alder ectomycorrhizae. The RFLP profiles of sporocarps and mycorrhizae were the same.

Agaricales↗

Carbon and nitrogen metabolism in ectomycorrhizal fungi and ectomycorrhizas.

The literature concerning the metabolism of carbon and nitrogen compounds in ectomycorrhizal associations of trees is reviewed. The absorption and translocation of mineral ions by the mycelia require an energy source and a reductant which are both supplied by respiratory catabolism of carbohydrates produced by the host plant. Photosynthates are also required to generate the carbon skeletons for amino acid and carbohydrate syntheses during the growth of the mycelia. Competition for photosynthates occurs between the fungal cells and the various vegetative sinks in the host tree. The nature of carbon compounds involved in these processes, their routes of metabolism, the mechanisms of control and the partitioning of metabolites between the various sites of utilization are only poorly understood. Both ascomycetous and basidiomycetous ectomycorrhizal fungi synthesize and some, if not all, accumulate mannitol, trehalose and triglycerides. The fungal strains employ the Embden--Meyerhof pathway of glucose catabolism and the key enzymes of the pentose phosphate pathway (6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, transaldolase and transketolase). Anaplerotic CO2 fixation, via pyruvate carboxylase and/or phosphoenolpyruvate carboxykinase, provides high pools of amino acids. This process could be important in the recapture and assimilation of respired CO2 in the rhizosphere. The ectomycorrhizas are thought to contain the Embden--Meyerhof pathway, the pentose phosphate pathway and the tricarboxylic acid cycle, which provide the carbon skeletons for the assimilation of ammonia into amino acids. The main route of assimilation of ammonia appears to be through the glutamine synthetase-glutamate synthase cycle in the ectomycorrhizas. Glutamate dehydrogenase plays a minor role in this process. Glutamate dehydrogenase and glutamine synthetase are present in free-living ectomycorrhizal fungi and they participate in the assimilation of ammonia and the synthesis of amino acids through the glutamate dehydrogenase/glutamine synthetase sequence. In both in vitro cultures of fungi and ectomycorrhizas, the assimilated nitrogen accumulates in glutamine. Glutamine, but also ammonia, are thought to be exported from the fungal tissues to the host cells. Studies on the metabolism of ectomycorrhizas and ectomycorrhizal fungi have focused on the metabolic pathways and compounds which accumulate in the symbiotic tissues. Studies on regulation of the overall process, and the control of enzyme activity in particular, are still fragmentary.(ABSTRACT TRUNCATED AT 400 WORDS)

Carbon↗

A novel class of ectomycorrhiza-regulated cell wall polypeptides in Pisolithus tinctorius.

Development of the ectomycorrhizal symbiosis leads to the aggregation of fungal hyphae to form the mantle. To identify cell surface proteins involved in this developmental step, changes in the biosynthesis of fungal cell wall proteins were examined in Eucalyptus globulus-Pisolithus tinctorius ectomycorrhizas by two-dimensional polyacrylamide gel electrophoresis. Enhanced synthesis of several immunologically related fungal 31- and 32-kDa polypeptides, so-called symbiosis-regulated acidic polypeptides (SRAPs), was observed. Peptide sequences of SRAP32d were obtained after trypsin digestion. These peptides were found in the predicted sequence of six closely related fungal cDNAs coding for ectomycorrhiza up-regulated transcripts. The PtSRAP32 cDNAs represented about 10% of the differentially expressed cDNAs in ectomycorrhiza and are predicted to encode alanine-rich proteins of 28.2 kDa. There are no sequence homologies between SRAPs and previously identified proteins, but they contain the Arg-Gly-Asp (RGD) motif found in cell-adhesion proteins. SRAPs were observed on the hyphal surface by immunoelectron microscopy. They were also found in the host cell wall when P. tinctorius attached to the root surface. RNA blot analysis showed that the steady-state level of PtSRAP32 transcripts exhibited a drastic up-regulation when fungal hyphae form the mantle. These results suggest that SRAPs may form part of a cell-cell adhesion system needed for aggregation of hyphae in ectomycorrhizas.

Amino Acid Sequence↗

Russulaceae and Thelephoraceae form ectomycorrhizas with members of the Nyctaginaceae (Caryophyllales) in the tropical mountain rain forest of southern Ecuador.

* Three members of the Nyctaginaceae, two Neea species and one Guapira species, occurred scattered within a very species-rich neotropical mountain rain forest. The three species were found to form ectomycorrhizas of very distinctive characters, while all other tree species examined formed arbuscular mycorrhizas. * The ectomycorrhizas were structurally typified according to light and transmission electron microscope investigations. The internal transcribed spacer (ITS) rDNA and part of the nuclear large subunit (LSU, 28S) rDNA of the mycorrhiza forming fungi were amplified and sequenced. Phylogenetic analyses were carried out. * Neea species 1 was found to form typical ectomycorrhizas with five different fungal species, Russula puiggarii, Lactarius sp., two Tomentella or Thelephora species, and one ascomycete. Neea species 2 and the Guapira species were associated with only one fungus each, a Tomentella/Thelephora species clustering closely together in an ITS-neighbour-joining tree. The long and fine rootlets of the Guapira species showed proximally a hyphal mantle and a Hartig net, but distally intracellular fungal colonization of the epidermis and root hair development. The ectomycorrhizal segments of the long roots of Neea species 2 displayed a hyphal mantle and a Hartig net around alive root-hair-like outgrowths of the epidermal cells. * The distribution and the evolution of ectomycorrhizas in the predominantly neotropic Nyctaginaceae are discussed.

Ascomycota↗

Soil bacterial diversity in a loblolly pine plantation: influence of ectomycorrhizas and fertilization.

We studied the effect of ectomycorrhizas and fertilization on soil microbial communities associated with roots of 10-year-old loblolly pine. Ectomycorrhizas were identified using a combination of community terminal restriction fragment profiling and matching of individual terminal restriction fragments to those produced from ectomycorrhizal clones and sequences recovered from roots and sporocarps. Differences between bacterial communities were initially determined using cluster analysis on community terminal restriction fragment profiles and through subsequent recovery of 16S rDNA clones. Analysis of bacterial clones revealed that terminal restriction fragment length was often shared between taxonomically dissimilar bacterial types. Consequently, we could not reliably infer the identity of peaks in the bacterial community profile with some exceptions, notably chloroplast rDNA that generated an approximate peak size of 80.2 bp. Fertilization increased the frequency of a Piloderma-like ectomycorrhiza. However, we did not detect clear effects of fertilization or the presence of viable ectomycorrhizas on bacterial communities. Bacterial communities seemed to be determined largely by the carbon and nitrogen content of soil. These results suggest that important soil microbial groups respond differently to soil conditions and management practices, with ectomycorrhizal communities reflecting past nutrient conditions and bacterial communities reflecting current environmental conditions of soil microsites.

Agriculture↗

Headspace solid-phase microextraction with gas chromatography and mass spectrometry in the investigation of volatile organic compounds in an ectomycorrhizae synthesis system.

Ectomycorrhizae formation represents one of the most significant steps in the truffle life cycle and is determined by a complex molecular signaling between two symbionts. In order to understand the molecular pathway of ectomycorrhiza development, we focused on the signaling interaction between the ectomycorrhizal fungus Tuber borchii Vittad. and the Tilia americana L. plant roots. The medium of a pre-symbiotic (T. americana-T. borchii) in vitro system was analysed by headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. In total, 73 volatile organic compounds (VOCs) were identified. Twenty-nine of these VOCs were produced only during the interaction phase between the two partners, leading to a hypothesis that these molecules might act as molecular messengers in order to pilot the ectomycorrhizae formation.

Gas Chromatography-Mass Spectrometry↗

Dispersed polyphosphate in fungal vacuoles in Eucalyptus pilularis/Pisolithus tinctorius ectomycorrhizas.

Ectomycorrhizas produced between Pisolithus tinctorius and Eucalyptus pilularis under axenic conditions were rapidly frozen, freeze-substituted in tetrahydrofuran and embedded anhydrously, and dry-sectioned for X-ray microanalysis. The vacuoles of the sheath and Hartig net hyphae were rich in phosphorus and potassium. They also contained sulfur and variable amounts of chlorine. In anhydrously processed freeze-substituted mycorrhizas, dispersed electron-opaque material filled the fungal vacuoles. X-ray maps indicated that P was distributed evenly throughout the entire vacuole profile and was not concentrated in spherical bodies or subregions of the vacuole. There were no electron-opaque granules surrounded by electron-lucent areas, such as are commonly seen in chemically fixed material. The fungal vacuoles were also rich in K, which similarly gave a signal from the entire vacuolar profile. Such P-rich vacuoles occurred in both the mycorrhizal sheath and Hartig net hyphae. Stained sections of ether-acrolein freeze-substituted mycorrhizas also showed only dispersed material in the fungal vacuoles as, in most cases, did acetone-osmium freeze-substituted material. Precipitation of metachromatic granules by ethanol suggested that large amounts of polyphosphate are stored in these regions under the conditions of our experiments, as well as in the tips of actively growing hyphae of the same fungus. The higher plant vacuoles of ectomycorrhizas gave a much lower signal for K, and P was barely detectable. Much more K was located in the vacuoles of the root exodermal cells than in epidermal cells. The analysis of element distribution between the vacuole and cytoplasm in root cells agrees well with that found for other plant species using other techniques. We conclude that polyphosphate is indeed present in the vacuoles of the fungal cells of these ectomycorrhizas, but that in vivo it is in a dispersed form, not in granules.

Basidiomycota↗

Ectomycorrhizas of Cortinarius helodes and Gyrodon monticola with Alnus acuminata from Argentina.

Field ectomycorrhizas of Cortinarius helodes Moser, Matheny & Daniele (sp. nov) and Gyrodon monticola Sing. on Alnus acuminata Kunth (Andean alder, aliso del cerro) are described based on morphological and anatomical features. Ectomycorrhizal roots were sampled beneath fruitbodies of C. helodes and G. monticola from two homogeneous A. acuminata forest sites located in Tucuman and Catamarca Provinces in Argentina. C. helodes ectomycorrhizas showed a thick white to beige mantle exuding a milky juice when injured, were bluish toward the apex, and had hyphal strands in the mantle. G. monticola ectomycorrhizas showed some conspicuous features like highly differentiated rhizomorphs, inflated brown cells on the mantle surface, and hyaline and brown emanating hyphae with dolipores. Restriction fragment length polymorphism analysis of the nuclear rDNA internal transcribed spacer provided a distinctive profile for each of the collections of fruitbodies and the mycorrhizal morphotypes.

Agaricales↗

Nutrient amounts of ectomycorrhizae analysed by EDX using ESEM and ICP.

Energy-dispersive X-ray (EDX) analysis coupled with an environmental scanning electron microscope (ESEM) was tested as an alternative to the inductively coupled argon plasma (ICP) spectrometer method for nutrient analyses of ectomycorrhizae. The results of EDX-ESEM and ICP were compared for 12 ectomycorrhizal morphotypes collected in beech and Scots pine forests in northern Brandenburg. The amounts of Al, Ca, Mg and S analysed in the outer hyphal layers of the sheath with the EDX-ESEM technique correlated well with the amounts of these elements in the whole mycorrhiza as assessed by ICP. For the elements P and K, no such correlation existed, indicating an uneven distribution of these elements in the ectomycorrhiza. It is concluded that the EDX-ESEM technique could be a useful and reliable tool for the analysis of nutrient elements in ectomycorrhizae, especially for studies focussing on small-scale soil heterogeneity or on infrequent morphotypes.

Electron Probe Microanalysis↗

Morphological and molecular comparison of white truffle ectomycorrhizae.

In the present study, white truffle ectomycorrhizae (EM) collected in deciduous forests (Populus, Quercus, and Fagus) from Hungary were characterized by morphological-anatomical and molecular methods. Our investigations suggest that the EM of white truffles (e.g., Tuber rapaeodorum, Tuber puberulum, Tuber rufum) are common and abundant members of the forest communities in the area. The ITS sequences of 14 EM specimens and 46 additional fruitbody sequences from the GenBank were clustered into four main groups in phylogenetic analyses. In the ITS-1 region, a characteristic indel pattern was found, which supports the clades. Although our analyses indicate definite genetic distance between the groups of the phylogenetic tree, these clades do not correspond to the traditional taxons identified by fruitbody characteristics. Comparison of the ectomycorrhizae shows that neither is mycorrhizal anatomy a good tool to separate the groups, because the characters (like the epidermoid or angular mantle structure, cell wall thickness, the sape and size of cystidia) are too variable and overlap between the clades. The interspecific similarity, observed both in ectomycorrhizal and fruitbody characters, strengthen the sensu lato morpho-species concept of this group. Our study, which combines comprehensive molecular and anatomical approach to characterize and identify ectomycorrhizae of white truffles from natural samples, stress out the need of the taxonomical revision of this group.

Basidiomycota↗

Growth, ectomycorrhizae and nonstructural carbohydrates of loblolly pine seedlings exposed to ozone and soil water deficit.

Loblolly pine (Pinus taeda) seedlings from three full-sib families were exposed to 0, 50, 100 or 150 ppb ozone (O(3)) (5 h/d, 5 d/week for 6 or 12 weeks). Soil water potential was maintained near pot capacity (-0.03 MPa) or soil was allowed to dry to approximately -1.0 MPa and resaturated. Chlorotic mottling and flecking of needles due to O(3) injury were observed for seedlings from all pine families. Soil water deficit lessened the intensity of O(3) symptoms, possibly due to stomatal closure. Exposure to O(3) and soil water deficit each resulted in less seedling volume growth and dry weight, and changed the nonstructural carbohydrate content of seedlings compared with controls. Increasing O(3) concentrations resulted in a linear reduction in foliar starch content but did and affect hexose or sucrose content. Soil water deficit resulted in less starch and soluble sugar contents in above- and below-ground plant parts compared with controls. Soil water deficit did not affect numbers or percentages of roots that formed ectomycorrhizal tips. A linear dose-response relationship between O(3) and ectomycorrhizae was observed. The number of ectomycorrhizal tips/cm long root and the percentage of feeder roots that formed ectomycorrhizae were lower as O(3) concentration increased. Overall, each stress alone caused less seedling growth and carbohydrate content compared with controls, but only O(3) was responsible for suppression of ectomycorrhizae.

Journal Article↗