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Humus quality after eucalypt reforestations in Asturias (Northern Spain).

The impact on soil properties of recent and ancient reforestations with Eucalyptus globulus Labill. has been studied in Cambisols and Leptosols under temperate semi-oceanic climate in Asturias (Northern Spain). Eucalypt forests showed a high potential of C accumulation in soil, mainly the ancient forest, where the organic matter concentration in the uppermost horizon (up to 415 C g x kg(-1)) was greater than in all neighbouring soil formations (climacic oak forests, up to 84 C g x kg(-1); Podsol under reforested pine forests, 287 C g x kg(-1); and Ulex shrub, 70 C g x kg(-1)). Nevertheless, there was a low concentration of available bases and a very low transformation degree of the organic matter in this eucalypt forest, only comparable to that of the Podsol. In order to analyse the major features of the biogeochemical behaviour of the sites under study, the humus fractions were isolated and the humic acid fraction was studied by derivative visible and infrared spectroscopies. Incubation experiments on whole soil samples were carried out to monitor stability against biodegradation of the organic matter under comparable conditions. In general, soils under eucalypt had a CO(2) production similar to or lower than the control sites, but the soil under the ancient eucalypt forest had a high potential for CO(2) release, similar to that of the neighbouring Podsol, what might be explained by the presence in these soils of high amounts of C accumulated in readily biodegradable forms. However, the mineralisation coefficients (CO(2) released per unit of soil C) indicated that the biodegradability of the soil humus has decreased significantly in the eucalypt-reforested sites, as it could correspond to an effective control of the biogeochemical processes caused by the antimicrobial products from the eucalypt litter. Compared to the original deciduous forests, the lower values of the E(4)/E(6) ratio in the humic acids from eucalypt forests and the lignin signature in the infrared spectra point to humification mechanisms based on the selective preservation of complex macromolecular substances derived from vascular plants. The second derivative of the visible spectra of the humic acid samples under study showed conspicuous valleys, which are ascribed to fungal-derived 4,9-dihydroxyperylene-3,10-quinone pigments. These valleys were in most eucalypt forests significantly shallower than in the original deciduous forests and may also represent an indicator of the impact of introduced vegetation on the structure of the soil microbial system and the mechanisms responsible for the formation of humic acids.

Carbon Dioxide↗

Does leaf water efficiency vary among eucalypts in water-limited environments?

There is a need to generalize water use behavior of eucalypts to facilitate bioengineering and landscape remediation programs in a wide range of Australian environments. A critical question can be stated as a null hypothesis: tree water use per unit leaf area (leaf efficiency) is independent of eucalypt species. This is implicitly equivalent to the hydrological equilibrium hypothesis that states that leaf area is a function of climate, at least in cases where transpiration and growth are limited by soil water. Failure to reject this null hypothesis simplifies (a) the selection of tree species for water balance management, (b) the generation of regional-scale expectations of leaf area index, and (c) the estimation (monitoring) of the effectiveness of plantations in controlling site water balance. The hypothesis was tested with tree water use data collected in natural multi-species stands across Australia, including sites in the wet-dry season tropical woodlands of the Northern Territory, the Mediterranean climate forests of Western Australia, and a woodland system in southern New South Wales receiving an even distribution of rainfall throughout the year. We also tested the hypothesis in a multi-species tree plantation growing on a saline gradient. In each case, we could not reject the hypothesis of constant leaf efficiency among eucalypts. In every case there was a common, strong, linear relationship among tree leaf area and mean daily water use by all tree species in a sample. Single factor (species) analysis of variance did not detect significant differences between leaf water efficiencies of species. For the jarrah forest (Eucalyptus marginata J. Donn ex Sm., E. calophylla R. Br. ex Lindl.), the null hypothesis held in both spring (wet) and autumn (dry) conditions. The null hypothesis held in the mixed species woodland of New South Wales (E. macrorhynca F.J. Muell. ex Benth., E. blakelyi Maiden., E. polyanthemos Schauer.) under summer and autumn conditions, and across five species in the wet-dry tropical woodland (E. miniata A. Cunn. ex Schauer, E. tetrodonta F.J. Muell., E. porecta S.T. Blake, Erythrophleum chlorostachys F.J. Muell., and Terminalia ferdinandiana Exell.). The null hypothesis also held for a plantation of E. occidentalis Endl. and provenances of E. camaldulensis Dehnh. growing on a shallow saline gradient; i.e., leaf water efficiency remained constant across species and varieties despite obvious effects of salinity on the size of individual canopies. We conclude that there is little evidence for rejecting the hypothesis that leaf efficiency does not vary significantly among sympatric eucalypt species in rainfall-limited (soil-water-limited) systems. These findings open the way for useful bioengineering generalities about the hydrological role of trees in the Australian landscape.

Journal Article↗

Responses of eucalypt species to aluminum: the possible involvement of low molecular weight organic acids in the Al tolerance mechanism.

Aluminum (Al) tolerance mechanisms in crop plants have been extensively researched, but our understanding of the physiological mechanisms underlying Al tolerance in trees is still limited. To investigate Al tolerance in eucalypts, seedlings of six species (Eucalyptus globulus Labill., Eucalyptus urophylla S.T. Blake, Eucalyptus dunnii Maiden, Eucalyptus saligna Sm., Eucalyptus cloeziana F. J. Muell. and Eucalyptus grandis w. Hill ex Maiden) and seedlings of six clones of Eucalyptus species were grown for 10 days in nutrient solutions containing Al concentrations varying from 0 to 2.5 microM (0 to 648 microM Al3+ activities). Root elongation of most species was inhibited only by high Al3+ activities. Low to intermediate Al3+ activities were beneficial to root elongation of all species and clones. Among the species tested, E. globulus and E. urophylla were more tolerant to Al toxicity, whereas E. grandis and E. cloeziana were more susceptible to Al-induced damage. Although E. globulus seedlings were tolerant to Al toxicity, they were highly sensitive to lanthanum (La), indicating that the tolerance mechanism is specific for Al. Fine roots accumulated more Al and their elongation was inhibited more than that of thick roots. In E. globulus, accumulation of Al in root tips increased linearly with increasing Al concentration in the nutrient solution. The majority of Al taken up was retained in the root system, and the small amounts of Al translocated to the shoot system were found mainly in older leaves. No more than 60% of the Al in the thick root tip was in an exchangeable form in the apoplast that could be removed by sequential citrate rinses. Gas chromatography/mass spectrometry and ion chromatography analyses indicated that root exposure to Al led to a greater than 200% increase in malic acid concentration in the root tips of all eucalypt species. The increase in malate concentration in response to Al treatment correlated with the degree of Al tolerance of the species. A small increase in citric acid concentration was also observed in all species, but there were no consistent changes in the concentrations of other organic acids in response to Al treatment. In all eucalypt species, Al treatment induced the secretion of citric and malic acid in root exudates, but no trend with respect to Al tolerance was observed. Thus, although malate and citrate exudation by roots may partially account for the overall high Al tolerance of these eucalypt species, it appears that tolerance is mainly derived from the internal detoxification of Al by complexation with malic acid.

Aluminum↗

Eucalypt MADS-box genes expressed in developing flowers.

Three MADS-box genes were identified from a cDNA library derived from young flowers of Eucalyptus grandis W. Hill ex Maiden. The three egm genes are single-copy genes and are expressed almost exclusively in flowers. The egm1 and egm3 genes shared strongest homology with other plant MADS-box genes, which mediate between the floral meristem and the organ-identity genes. The egm3 gene was also expressed strongly in the receptacle or floral tube, which surrounds the carpels in the eucalypt flower and bears the sepals, petals, and numerous stamens. There appeared to be a group of genes in eucalypts with strong homology with the 3' region of the egm1 gene. The egm2 gene was expressed in eucalypt petals and stamens and was most homologous to MADS-box genes, which belong to the globosa group of genes, which regulate organogenesis of the second and third floral whorls. The possible role of these three genes in eucalypt floral development is discussed.

Amino Acid Sequence↗

[Effects of continuous cropping on understorey species diversity in Eucalypt plantations].

By the methods of time-space series analysis and field monitoring, this paper studied the species diversity and its maintaining mechanism in Dongmen Eucalypt plantations of Guangxi. The results indicated that during the period of 1998 - 2003, continuous cropping of Eucalyptus plantation resulted in a reduction of species diversity. The plant species number of second generation was reduced by 54.43% in a 667 m2 plot, and by 50% in 18 sampling plots (each of 4 m2), and the species richness and Shannon-Wiener index of 6-year second generation forest decreased by 39.39% and 17.76%, respectively, compared with the first generation. The continuous cropping not only decreased the species diversity, but also changed the species composition and characteristics of the community. Life-form spectra of Eucalypt plantations varied with generation. The plant life-form consisted of microphanerophytes, liana phanerophytes, and nana-phanerophytes in the first generation, while the herbaceous-phanerophytes and chamaephytes occurred in the second generation. The k-strategy and r-strategy species accounted for 69.32% and 30.68% in the first generation, while 47.83% and 52.17% in the second generation, respectively. Therefore, continuous cropping of Eucalypt plantations was advantage for herb, therophytes and r-strategy species, but disadvantage for woody plants and k-strategy species. Seed band and vegetative body in soil were considered to play an important role in maintaining diversity. A new hypothesis, initial plant Diaspora, on the maintaining mechanism of species diversity in Eucalypt plantation was put forward.

Biodiversity↗

Gas chromatography/mass spectrometry demonstration of steryl glycosides in eucalypt wood, Kraft pulp and process liquids.

The occurrence of steryl glycosides (SG) and acyl steryl glycosides (ASG) in eucalypt (Eucalyptus globulus) wood has been investigated. These compounds were analyzed as their trimethylsilyl ethers by gas chromatography/mass spectrometry (GC/MS) using a 15 m length high-temperature capillary column with a thin film, and identified on basis of their mass spectra and relative retention times comparing with those of authentic standards. Significant amounts of SG were identified in eucalypt wood whilst only traces of ASG could be detected. Eucalypt SG and ASG occur in the pyranoside form, which is readily distinguishable from the furanoside configuration by the mass spectra of their trimethylsilyl derivatives. The sterol part of the SG and ASG consisted of sitosterol, being sitosteryl 3beta-D-glucopyranoside and sitosteryl (6'-O-palmitoyl)-3beta-D-glucopyranoside, the major SG and ASG found in E. globulus wood. The presence of SG and ASG was also investigated after kraft cooking by analyzing unbleached pulp and process water samples. The GC/MS results also revealed the presence of sitosteryl 3beta-D-glucopyranoside in these samples. By contrast, no ASG could be detected. Therefore, we have shown that SG survive the kraft cooking and can be found at least partly intact after pulping, being a possible cause for pitch deposits together with free and esterified sterols.

Acylation↗

Enzymatic removal of free and conjugated sterols forming pitch deposits in environmentally sound bleaching of eucalypt paper pulp.

Free and conjugated sterols are among the main compounds responsible for pitch deposition in the manufacture of wood chemical pulps, making difficult the implementation of totally chlorine free bleaching (TCF) and closure of bleach plant circuits. In this work, the suitability of oxidative enzymes in efficiently removing sterols from eucalypt pulps is revealed. The enzymatic treatment was applied as an additional stage of an industrial-type TCF sequence for bleaching eucalypt kraft pulp. The pulp obtained after oxygen delignification was treated with a high-redox potential and thermostable fungal laccase using 1-hydroxybenzotriazole as an enzyme mediator. This pulp was further submitted to chelation and peroxide stages and compared with a control TCF pulp obtained using chemical reagents. The composition of the lipophilic extractives in the pulps and the corresponding liquids after the different stages was analyzed by gas chromatography and gas chromatography--mass spectrometry. Free sitosterol and sitosterol esters and glucosides, the major lipophilic compounds in eucalypt pulps, were completely removed during the laccase-mediator treatment. Only some intermediate products from sitosterol oxidation remained after the laccase stage, as well as in the final pulp. Pulp brightness was also improved due to the simultaneous removal of lignin by the laccase-mediator treatment.

Chromatography, Gas↗

Isolation of Cryptococcus neoformans var. neoformans serotype D from Eucalypts in South Brazil.

Cryptococcus neoformans causes the second most common opportunistic infection in patients with AIDS. In Brazil, 4.5% of the AIDS-related opportunistic infections are caused by C. neoformans and all varieties are recognized as etiological agents of cryptococcosis. This pathogen is a ubiquitous environmental yeast, commonly associated with avian excreta and decaying wood, especially Eucalypt species. The aim of the present study was to search for C. neoformans in Eucalypts and analyze the genotypic diversity of the obtained isolates by RAPD and PCR fingerprinting. All obtained isolates have been C. neoformans var. neoformans, serotype D molecular type VNIV. Serotype D, was isolated from 3 (37.5%) out of 8 cities surveyed in the South Brazilian state Rio Grande do Sul. Nine (9%) out of 99 environmental samples were obtained from Eucalypt species, Eucalyptus calmadulensis and Eucalyptus tereticornis. Molecular analysis using RAPD and PCR-fingerprinting revealed very little genetic diversity in the obtained cryptococcal serotype D isolates. To our knowledge this is the first report of the isolation of serotype D from Eucalyptus trees in Brazil. More studies are required in order to establish the ecological significance of this finding.

Brazil↗

Chlorophyll content in eucalypt vegetation at the leaf and canopy scales as derived from high resolution spectral data.

The physiological status of forest canopy foliage is influenced by a range of factors that affect leaf pigment content and function. Recently, several indices have been developed from remotely sensed data that attempt to provide robust estimates of leaf chlorophyll content. These indices have been developed from either hand-held spectroradiometer spectra or high spectral resolution (or hyperspectral) imagery. We determined if two previously published indices (Datt 1999), which were specifically developed to predict chlorophyll content in eucalypt vegetation by remote sensing at the leaf scale, can be extrapolated accurately to the canopy. We derived the two indices from hand-held spectroradiometer data of eucalypt leaves exhibiting a range of insect damage symptoms. We also derived the indices from spectra obtained from high spectral and spatial resolution Compact Airborne Spectrographic Imager 2 (CASI-2) imagery to determine if reasonable estimates at a scale of < 1 m can be achieved. One of the indices (R 850/R 710 index, where R is reflectance) derived from hand-held spectroradiometer data showed a moderate correlation with relative leaf chlorophyll content (r = 0.59, P < 0.05) for all dominant eucalypt species in the study area. The R (850)/R (710) index derived from CASI-2 imagery yielded slightly lower correlations over the entire data set (r = 0.42, P < 0.05), but correlations for individual species were high (r = 0.77, P < 0.05). A scaling analysis indicated that the R (850)/R (710) index was strongly affected by soil and water cover types when pixels were mixed, but appeared to be invariant to changes in proportions of understory, which may limit its application.

Chlorophyll↗

Metabolites of chlorinated syringaldehydes in fish bile as biomarkers of exposure to bleached eucalypt pulp effluents.

Metabolites of chlorinated phenolic compounds in fish bile have been found to be sensitive biomarkers of bleached pulp mill effluent exposure. Chlorinated syringaldehydes are largely unstudied chlorophenolics found in bleached hardwood effluent. Sand flathead (Platycephalus bassensis), Australian marine fish, were exposed to 100% chlorine dioxide-bleached eucalypt pulp effluent at concentrations of 0.5, 2, and 8% (v/v) for 4 days. Metabolites of 2-chlorosyringaldehyde (2-CSA), the predominant chlorophenolic in this effluent, were measured in the bile. The major metabolite was the conjugate of 2-chloro-4-hydroxy-3,5-dimethoxy-benzylalcohol (2-CB-OH), the reduced product of 2-CSA. 2-CB-OH was found in all fish exposed to diluted effluent and was concentrated in the bile over 1000 times above 2-CSA levels in the effluent. A separate experiment examined the metabolic fate of 2,6-dichlorosyringaldehyde (2,6-DCSA), which is one of the major chlorophenolics in chlorine-bleached eucalypt pulp effluent. Sand flathead were exposed to 2,6-DCSA by intraperitoneal injection at 15 mg/kg or through the water to 0.5, 2, or 8 micrograms/liter for 4 days. Analysis of the bile revealed the major metabolite of 2,6-DCSA to be the conjugate of 2,6-dichloro-4-hydroxy-3,5-dimethoxybenzylalcohol, which was found in all exposed fish and was concentrated in the bile over 20,000 times above 2,6-DCSA exposure levels. Results reveal that the analysis of metabolites of chlorinated syringaldehydes in fish bile can provide a biomarker of bleached hardwood effluent exposure that is sensitive to low levels of exposure, specific to certain bleaching sequences, and correlates well with exposure concentrations.

Animals↗

Lipophilic extractives in process waters during manufacturing of totally chlorine free kraft pulp from eucalypt wood.

The chemical composition of lipophilic extractives from process waters throughout a totally chlorine free (TCF) bleaching sequence after kraft pulping of eucalypt wood has been studied. These compounds are among the most problematic wood constituents for both TCF and zero liquid effluent (ZLE) processes, since they tend to accumulate in circuits resulting in the formation of the so-called pitch deposits causing serious problems in the pulp and paper industry. Pitch deposits collected at different parts of the pulp mill were also characterized and their composition compared with that of lipophilic compounds in process waters and Eucalyptus globulus wood. The identification of these compounds from process waters, wood and pitch deposits was performed by gas chromatography and gas chromatography-mass spectrometry using short- and medium-length high temperature capillary columns. Sterols, sterol esters, steroid ketones and steroid hydrocarbons were the main compounds identified. These chemical species arise from eucalypt wood extractives that survive the pulping and are released from pulp into the process waters during the bleaching process. Finally, they can deposit in pulp and on different parts of the mill or remain suspended in process waters being discharged in effluents.

Chromatography, Gas↗

Soil response to eucalypt tree planting and meatworks effluent irrigation in a short rotation forest regime in New Zealand.

The effects of planting three eucalypt species and irrigating with meatworks effluent on soil were assessed during the first 3-year rotation of a short rotation forest regime at Oringi, Dannevirke, New Zealand. The results showed tree planting alone reduced the soil infiltration rates, but had little influence on soil nutrient concentration other than reduction of nitrate levels. Species variation had limited influence on soil change. Effluent irrigation relieved the reduction of infiltration rates by tree planting, and increased nutrient concentrations, but reduced the soil pH. These changes should be considered when managing eucalypt short rotation forests sustainably in the longer term, either linked with effluent irrigation or not.

Agriculture↗

Insect pests of eucalypts in California: implications of managing invasive species.

For the first 150 years following their introduction, eucalypts planted in the California landscape were free of both insect pests and diseases. In the last 15 years, numerous herbivorous insect species have been introduced accidentally into the State and have caused significant damage to the trees. Several of these species, e.g. Phoracantha semipunctata (Fabricius), Phoracantha recurva Newman (Coleoptera: Cerambycidae) and Gonipterus scutellatus Gyllenhal (Coleoptera: Curculionidae), have also been introduced into other parts of the world where eucalypts are grown, whereas others, e.g. Glycaspsis brimblecombei Moore (Hemiptera: Spondyliaspidae) and Eucalyptolyma maideni Froggatt (Hemiptera: Spondyliaspidae), are currently restricted to California and Australia. Research programmes have provided management solutions to individual pest problems, but as more pest species are introduced, these solutions must be integrated across broad geographic, horticultural, and economic scales, in a systems approach.

Animals↗

Hypaphorine from the ectomycorrhizal fungus Pisolithus tinctorius counteracts activities of indole-3-acetic acid and ethylene but not synthetic auxins in eucalypt seedlings.

Very little is known about the molecules regulating the interaction between plants and ectomycorrhizal fungi during root colonization. The role of fungal auxin in ectomycorrhiza has repeatedly been suggested and questioned, suggesting that, if fungal auxin controls some steps of colonized root development, its activity might be tightly controlled in time and in space by plant and/or fungal regulatory mechanisms. We demonstrate that fungal hypaphorine, the betaine of tryptophan, counteracts the activity of indole-3-acetic acid (IAA) on eucalypt tap root elongation but does not affect the activity of the IAA analogs 2,4-D ((2,4-dichlorophenoxy)acetic acid) or NAA (1-naphthaleneacetic acid). These data suggest that IAA and hypaphorine interact during the very early steps of the IAA perception or signal transduction pathway. Furthermore, while seedling treatment with 1-amincocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, results in formation of a hypocotyl apical hook, hypaphorine application as well as root colonization by Pisolithus tinctorius, a hypaphorine-accumulating ectomycorrhizal fungus, stimulated hook opening. Hypaphorine counteraction with ACC is likely a consequence of hypaphorine interaction with IAA. In most plant-microbe interactions studied, the interactions result in increased auxin synthesis or auxin accumulation in plant tissues. The P. tinctorius / eucalypt interaction is intriguing because in this interaction the microbe down-regulates the auxin activity in the host plant. Hypaphorine might be the first specific IAA antagonist identified.

Amino Acids↗

Glacial refugia and reticulate evolution: the case of the Tasmanian eucalypts.

Tasmania is a natural laboratory for investigating the evolutionary processes of the Quaternary. It is a large island lying 40-44 degrees S, which was repeatedly glaciated and linked to southeastern continental Australia during the Quaternary. Climate change promoted both the isolation of species in glacial refugia, and an exchange between Tasmanian and mainland floras. Eucalyptus is a complex and diverse genus, which has increased in abundance in Australia over the past 100 kyr, probably in response to higher fire frequency. Morphological evidence suggests that gene flow may have occurred between many eucalypt species after changes in their distribution during the Quaternary. This paper summarizes recent genetic evidence for migration and introgressive hybridization in Tasmanian Eucalyptus. Maternally inherited chloroplast DNA reveals a long-term persistence of eucalypts in southeastern Tasmanian refugia, coupled with introgressive hybridization involving many species. Detailed analysis of the widespread species Eucalyptus globulus suggests that migration from mainland Australia was followed by introgression involving a rare Tasmanian endemic. The data support the hypothesis that changes in distribution of interfertile species during the Quaternary have promoted reticulate evolution in Eucalyptus.

Climate↗

Ectomycorrhizin Synthesis and Polypeptide Changes during the Early Stage of Eucalypt Mycorrhiza Development.

In functioning eucalypt ectomycorrhizas, biochemical alterations are accompanied by a differential accumulation of polypeptides including the synthesis of symbiosis-related proteins (JL Hilbert, Martin FM [1988] New Phytol 110: 339-346). In the present study, protein biosynthesis in the early stages of ectomycorrhiza formation on Eucalyptus globulus subsp. bicostata Kirkp. was examined using compatible and incompatible isolates of the basidiomycete Pisolithus tinctorius (Coker & Couch). Changes in polypeptide composition were observed within hours following contact of the compatible mycelium with the roots, well before the differentiation of typical symbiotic tissues. At this stage, at least seven symbiosis-related proteins (ectomycorrhizins) accumulated in root tissues. In vivo incorporation of [(35)S]methionine by ectomycorrhizas followed by electrophoresis of the labeled proteins revealed that most of these differences in polypeptide concentrations, including the ectomycorrhizin accumulation, are the result of differential protein biosynthesis rather than posttranslational modifications of the polypeptides. The initial development of eucalypt ectomycorrhizas, therefore, coincides with the synthesis of symbiosis-related proteins and the data presented here provide essential evidence to ascribe a functional developmental role to these proteins.

Journal Article↗

Assessment of crown condition in eucalypt vegetation by remotely sensed optical indices.

Leaf and crown damage and discoloration characteristics are important variables when defining the health of eucalypt tree species and have been used as key indicators of environmental quality. These indicators can vary significantly over a few hectares, especially in mixed-species forests, making field-based environmental surveillance of crown condition an extremely expensive and logistically impractical task. Reflectance in narrow spectral wavelengths obtained from a field-based spectroradiometer and a Compact Airborne Spectrographic Imager 2 (CASI-2) were collected over eucalypt vegetation of varying condition in southeastern Australia and compared with leaf- and crown-based attributes including percent red foliage discoloration, percent leaf damage, and crown density and crown foliage condition. Of the leaf attributes sampled, percent leaf damage was well correlated with a red-green spectral index (r = 0.68, p < 0.01), and percent red discoloration was well correlated with the slope of the red-edge for selected species (r = 0.89, p < 0.001). Within-tree crown density was well correlated with the slope of the red-edge (r = 0.77, p < 0.001) and a previously published index of plant stress with crown foliage condition (r = 0.88, p < 0.01) for selected species. Despite evidence of strong interspecific variability, a set of narrow spectral wavelengths in the visible and near-infrared regions of the spectrum have been identified that will be useful in the development of forest ecosystem environmental quality indicators.

Ecosystem↗

The expression of a symbiosis-regulated gene in eucalypt roots is regulated by auxins and hypaphorine, the tryptophan betaine of the ectomycorrhizal basidiomycete Pisolithus tinctorius.

A full-length cDNA coding for a symbiosis-regulated transcript, EgHypar, was isolated by differential screening from a Eucalyptus globulus bicostata--Pisolithus tinctorius ectomycorrhiza. The sequence of this clone revealed a protein with an estimated molecular mass of 25.5 kDa that exhibited a high degree of homology (66%) with plant auxin-induced glutathione-S-transferases. Expression of the EgHypar gene in seedlings was confined largely in roots and it is drastically increased by ectomycorrhiza development. The concentration of EgHypar transcripts was similarly up-regulated in roots incubated in media supplemented with P. tinctorius cell-free extracts, indole-3-acetic acid, 2,4-dichlorophenoxyacetic acid or hypaphorine (tryptophan betaine), the major indolic compound secreted by P. tinctorius. The latter fungal alkaloid concomitantly induced a decrease in root hair elongation in eucalypt seedlings. Up-regulation of EgHypar expression by auxins and fungal metabolites suggests that this symbiosis-regulated gene could be involved in the morphological changes taking place in plants roots upon symbiosis development. To our knowledge, these results provide the first molecular evidence that gene expression of the host plant is altered by molecules produced by the ectomycorrhizal mycobiont.

Amino Acid Sequence↗