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genetic control of eucalyptus urophylla and e. grandis resistance to canker caused by chrysoporthe cubensis.chrysophorte cubensis induced canker occurs in nearly all tropical and subtropical regions where eucalypts are planted, causing losses in both wood quality and volume productivity, especially so in the warmer and more humid regions of brazil. the wide inter and intra-specific genetic variability of resistance to canker among eucalyptus species facilitates the selection of resistant plants. in this study, we evaluated resistance to this pathogen in five eucalyptus grandis (g) and 15 e. urophylla ...201021637427
effect of overexpression of radish plasma membrane aquaporins on water-use efficiency, photosynthesis and growth of eucalyptus trees.eucalyptus is a diverse genus of flowering trees with more than 700 genotypic species which are mostly native to australia. we selected 19 wild provenances of eucalyptus camaldulensis grown in australia, compared their growth rate and drought tolerance and determined the protein levels of plasma membrane aquaporins (pips). there was a positive relationship between the drought tolerance and pip content. pips are divided into two subgroups, pip1 and pip2. most members of the pip2 subgroup, but not ...201020124554
identification of differentially expressed genes of the fungus hydnangium sp. during the pre-symbiotic phase of the ectomycorrhizal association with eucalyptus grandis.the pre-symbiotic phase that precedes physical contact between symbionts is a crucial phase in determining their compatibility, allowing the formation of the ectomycorrhiza. a subtractive cdna library representing the differentially expressed genes of the fungus hydnangium sp. in the pre-symbiotic phase was constructed using fungal mycelia obtained through the in vitro mycorrhization technique. the fungus was cultured in the presence of eucalyptus grandis roots, but with no contact between the h ...201020177718
the influence of pulping and washing conditions on the properties of eucalyptus grandis unbleached kraft pulps treated with chelants.the influence of different addition points of a chelating agent and a counter-ion exchange on the properties of eucalyptus grandis unbleached kraft pulps is studied. seven pulps were considered: two laboratory kraft pulps with or without the inclusion of the chelant dtpmpa (diethylene triamine penta (methylene phosphonic acid)), a mill kraft pulp and four mill pulps after ca(+2) or na(+) counter-ion exchange followed, or not, by washing with dtpmpa addition. the laboratory pulps required lower b ...201019880316
[effects of fertilization level on diurnal variation of gas exchange of young eucalyptus grandis leaf].different levels (0, 90, 180, and 270 g per tree) of compound fertilizer containing 15% n, 15% p2o5, and 15% k2o were applied to young eucalyptus grandis to study the diurnal variations of its leaf stomatal conductance (gs), intercellular co2 concentration (ci), net photosynthesis rate (pn), transpiration rate (tr), water use efficiency (wue), and vapor pressure deficit on leaf surface (vpdl) as well as the variation of leaf chlorophyll content, aimed to approach the relationships of e. grandis ...201021360993
characterization of a small cryptic plasmid from endophytic pantoea agglomerans and its use in the construction of an expression vector.a circular cryptic plasmid named ppaga (2,734 bp) was isolated from pantoea agglomerans strain ege6 (an endophytic bacterial isolate from eucalyptus). sequence analysis revealed that the plasmid has a g+c content of 51% and contains four potential orfs, 238(a), 250(b), 131(c), and 129(d) amino acids in length without homology to known proteins. the shuttle vector plgm1 was constructed by combining the ppaga plasmid with pgfpmut3.0 (which harbors a gene encoding green fluorescent protein, gfp), a ...201121637551
new records of the cryphonectriaceae from southern africa including latruncellus aurorae gen. sp. nov.the cryphonectriaceae accommodates some of the world's most important tree pathogens, including four genera known from native and introduced myrtales in africa. surveys in the past 3 y in southern africa have led to the discovery of cankers with fruiting structures resembling those of the cryphonectriaceae on trees in the myrtales in namibia, south africa, swaziland and zambia. these fungi were identified with morphological characteristics and dna sequence data. for the first time we report chry ...201121262988
glyphosate reduces urediniospore development and puccinia psidii disease severity on eucalyptus grandis.recent studies have shown the effects of glyphosate drift on decreasing rust intensity on eucalyptus grandis plants. however, the effects of the herbicide on puccinia psidii initial development are unknown. in this study the systemic action of glyphosate on rust severity was evaluated on eucalyptus plants maintained under greenhouse conditions. urediniospore germination and apressorium formation on detached leaves and on water agar medium, previously treated with glyphosate, were also evaluated.201121370395
correspondence between performance of eucalyptus spp trees selected from family and clonal tests.we examined the correspondence in performance between trees selected from a family test and their respective clones from a clonal test of eucalyptus. full-sib families were obtained from controlled pollination among individuals of eucalyptus grandis and between e. grandis and e. urophylla. the hybridizations did not follow a factorial scheme. the family tests were carried out at three locations in eun+ípolis and itabela counties, in bahia, brazil, in 2003. four hundred and ninety-seven high-perf ...201121732281
insecticidal activity of essential oils from eleven eucalyptus spp. and two hybrids: lethal and sublethal effects of their major components on blattella germanica.the aim of the current study was to evaluate the fumigant activity of the essential oils from 11 species of the genus eucalyptus and two of their hybrids on first instar of blattella germanica l. the fumigant activity and repellence of the four major monoterpene components of these essential oils also were tested. fumigant activity was evaluated by exposing nymphs to the vapors emitted by 50 microl of essential oil or monoterpene in a closed container. the lowest knockdown time 50% (kt50) values ...201121510210
[decomposition of eucalyptus-alder mixed litters and dynamics of soil faunal community].in order to understand the decomposition characteristics of eucalyptus (eucalyptus grandis) -alder (alnus formosana) mixed litters and the roles of soil fauna in the decomposition process, litter samples of the two tree species were collected from the suji town of leshan, sichuan province, and mixed with different proportions. the mixtures were put in 6-, 30-, and 260-mesh litterbags to investigate their mass loss and the dynamics of soil faunal community during the process of decomposition. dif ...201121774303
do changes in carbon allocation account for the growth response to potassium and sodium applications in tropical eucalyptus plantations?understanding the underlying mechanisms that account for the impact of potassium (k) fertilization and its replacement by sodium (na) on tree growth is key to improving the management of forest plantations that are expanding over weathered tropical soils with low amounts of exchangeable bases. a complete randomized block design was planted with eucalyptus grandis (w. hill ex maiden) to quantify growth, carbon uptake and carbon partitioning using a carbon budget approach. a combination of approac ...201122021011
molecular weight distributions and linkages in lignocellulosic materials derivatized from ionic liquid media.a novel and reproducible method is described for accurately determining the molecular weight distribution by size exclusion chromatography (sec) of whole lignocellulosic materials. this approach offers the opportunity to compare the molecular weight distributions of intact milled woods and its component fractions, lignins and holocelluloses, all from the same source, thus highlighting the potential of the technique and the contributions of the individual components to the chromatogram. the metho ...201121235206
population genetic analysis and phylogeny reconstruction in eucalyptus (myrtaceae) using high-throughput, genome-wide genotyping.a set of over 8000 diversity arrays technology (dart) markers was tested for its utility in high-resolution population and phylogenetic studies across a range of eucalyptus taxa. small-scale population studies of eucalyptus camaldulensis, eucalyptus cladocalyx, eucalyptus globulus, eucalyptus grandis, eucalyptus nitens, eucalyptus pilularis and eucalyptus urophylla demonstrated the potential of genome-wide genotyping with dart markers to differentiate species, to identify interspecific hybrids a ...201121310251
a high-density transcript linkage map with 1,845 expressed genes positioned by microarray-based single feature polymorphisms (sfp) in eucalyptus.abstract:201121492453
plants used for treating respiratory infections in rural maputaland, kwazulu-natal, south africa.traditional remedies are frequently used in treating various respiratory ailments, and are very important in the primary health care of the people living in rural maputaland, kwazulu-natal, south africa. novel information gathered from surveys like the present study is important in preserving indigenous knowledge.201121497646
transpiration of urban forests in the los angeles metropolitan area.despite its importance for urban planning, landscape management, and water management, there are very few in situ estimates of urban-forest transpiration. because urban forests contain an unusual and diverse mix of species from many regions worldwide, we hypothesized that species composition would be a more important driver of spatial variability in urban-forest transpiration than meteorological variables in the los angeles (california, usa) region. we used constant-heat sap-flow sensors to moni ...201121639035
an antimicrobial evaluation of plants used for the treatment of respiratory infections in rural maputaland, kwazulu-natal, south africa.abundant availability of medicinal plants in the study area offers low cost health care, but scientific validation is needed in order to lend credibility to the traditional use against respiratory infections.201222960551
[pontoscolex corethrurus (annelidae: oligochaeta) soil quality indicator in eucalyptus grandis (myrtacea) sites with slash and burn management].soil burning has been used in agricultural and forestry systems as a fundamental technique to clean the land and add some nutrients to the soil. in addition, earthworms are known to promote various soil functions since they contribute to aeration and organic matter and nutrients availability to other soil organisms. this study evaluated the effects of tropical forest crops management with presence-absence of eucalyptus grandis on earthworm population in huimanquillo, tabasco, mexico. three sites ...201223342509
production and carbon allocation in monocultures and mixed-species plantations of eucalyptus grandis and acacia mangium in brazil.introducing nitrogen-fixing tree species in fast-growing eucalypt plantations has the potential to improve soil nitrogen availability compared with eucalypt monocultures. whether or not the changes in soil nutrient status and stand structure will lead to mixtures that out-yield monocultures depends on the balance between positive interactions and the negative effects of interspecific competition, and on their effect on carbon (c) uptake and partitioning. we used a c budget approach to quantify g ...201222588515
[effects of eucalyptus grandis leaf litter decomposition on the growth and photosynthetic characteristics of cichorium intybus].a pot experiment was conducted to study the effects of eucalyptus grandis leaf litter during its early stage decomposition on the growth and the photosynthesis of cichorium intybus. each pot contained 12 kg soil mixed with different amounts of e. grandis leaf litter (30 g x pot(-1), a1; 60 g x pot(-1), a2; 90 g x pot(-1), a3; and 0 g x pot(-1), ck), and sowed with c. intybus. the growth indicators and the photosynthetic characteristics of c. intybus were measured after the third leaf of c. intyb ...201222489472
reference genes for high-throughput quantitative reverse transcription-pcr analysis of gene expression in organs and tissues of eucalyptus grown in various environmental conditions.interest in the genomics of eucalyptus has skyrocketed thanks to the recent sequencing of the genome of eucalyptus grandis and to a growing number of large-scale transcriptomic studies. quantitative reverse transcription-pcr (rt-pcr) is the method of choice for gene expression analysis and can now also be used as a high-throughput method. the selection of appropriate internal controls is becoming of utmost importance to ensure accurate expression results in eucalyptus. to this end, we selected 2 ...201223161857
validation of reference genes from eucalyptus spp. under different stress conditions.the genus eucalyptus consists of approximately 600 species and subspecies and has a physiological plasticity that allows some species to propagate in different regions of the world. eucalyptus is a major source of cellulose for paper manufacturing, and its cultivation is limited by weather conditions, particularly water stress and low temperatures. gene expression studies using quantitative reverse transcription polymerase chain reaction (qpcr) require reference genes, which must have stable exp ...201223148685
land use change in a temperate grassland soil: afforestation effects on chemical properties and their ecological and mineralogical implications.the current change in land use of grassland in the temperate region of south america is a process associated with the worldwide expansion of annual crops and afforestation with fast growing exotic species. this last cultivation has particularly been the subject of numerous studies showing its negative effects on soil (acidification, loss of organic matter and base cations, among others). however its effects on the mineral fraction are not yet known, as it is generally considered as one of the sl ...201223064181
genomic characterization of dart markers based on high-density linkage analysis and physical mapping to the eucalyptus genome.diversity arrays technology (dart) provides a robust, high throughput, cost-effective method to query thousands of sequence polymorphisms in a single assay. despite the extensive use of this genotyping platform for numerous plant species, little is known regarding the sequence attributes and genome-wide distribution of dart markers. we investigated the genomic properties of the 7,680 dart marker probes of a eucalyptus array, by sequencing them, constructing a high density linkage map and carryin ...201222984541
enhancement of enzymatic digestibility of eucalyptus grandis pretreated by naoh catalyzed steam explosion.naoh catalyzed steam explosion was applied to improve the enzymatic digestibility of eucalyptus grandis. the impregnation of wood chip with naoh solutions before steam explosion was carried out to investigate the effects of alkaline catalysis on solid recovery, enzymatic digestibility, and glucose recovery. the lignin removal was significantly affected by naoh as a catalyst and the glucose recovery increased with increased severity index. the use of naoh showed higher enzymatic digestibility tha ...201222939603
removal of nitrogen and phosphorus by eucalyptus and populus at a tertiary treated municipal wastewater sprayfield.various progenies of eucalyptus grandis and e. amplifolia, and clones of populus deltoides, were evaluated for plant removal of nitrogen (n) and phosphorus (p) for 26 months at a municipal waste spray field in north florida. tertiary treated wastewater containing 2.73 mg l(-1) nitrate n and 0.30 mg l(-1) total p was applied using sprinkler irrigation (93.8 m3 ha(-1) d(-1)) to fast growing trees utilized for bioenergy. eucalyptus amplifolia and e. grandis survived and grew very poorly as the resu ...201222908661
genetic dissection of growth, wood basic density and gene expression in interspecific backcrosses of eucalyptus grandis and e. urophylla.f1 hybrid clones of eucalyptus grandis and e. urophylla are widely grown for pulp and paper production in tropical and subtropical regions. volume growth and wood quality are priority objectives in eucalyptus tree improvement. the molecular basis of quantitative variation and trait expression in eucalypt hybrids, however, remains largely unknown. the recent availability of a draft genome sequence (http://www.phytozome.net) and genome-wide genotyping platforms, combined with high levels of geneti ...201222817272
stand-level patterns of carbon fluxes and partitioning in a eucalyptus grandis plantation across a gradient of productivity, in sao paulo state, brazil.wood production represents a large but variable fraction of gross primary production (gpp) in highly productive eucalyptus plantations. assessing patterns of carbon (c) partitioning (c flux as a fraction of gpp) between above- and belowground components is essential to understand mechanisms driving the c budget of these plantations. better knowledge of fluxes and partitioning to woody and non-woody tissues in response to site characteristics and resource availability could provide opportunities ...201222543478
microarray analysis revealed upregulation of nitrate reductase in juvenile cuttings of eucalyptus grandis, which correlated with increased nitric oxide production and adventitious root formation.the loss of rooting capability following the transition from the juvenile to the mature phase is a known phenomenon in woody plant development. eucalyptus grandis was used here as a model system to study the differences in gene expression between juvenile and mature cuttings. rna was prepared from the base of the two types of cuttings before root induction and hybridized to a dna microarray of e. grandis. in juvenile cuttings, 363 transcripts were specifically upregulated, enriched in enzymes of ...201222519851
thermal decomposition of wood: influence of wood components and cellulose crystallite size.the influence of wood components and cellulose crystallinity on the thermal degradation behavior of different wood species has been investigated using thermogravimetry, chemical analysis and x-ray diffraction. four wood samples, pinus elliottii (pie), eucalyptus grandis (eug), mezilaurus itauba (ita) and dipteryx odorata (dip) were used in this study. the results showed that higher extractives contents associated with lower crystallinity and lower cellulose crystallite size can accelerate the de ...201222306076
thermal decomposition of wood: kinetics and degradation mechanisms.the influence of wood components and cellulose crystallinity on the kinetic degradation of different wood species has been investigated using thermogravimetry. four wood species were studied: pinus elliottii (pie), eucalyptus grandis (eug), mezilaurus itauba (ita) and dipteryx odorata (dip). thermogravimetric results showed that higher extractive contents in the wood accelerate the degradation process and promote an increase in the conversion values at low temperatures. alternatively, the result ...201223073083
induced somatic sector analysis of cellulose synthase (cesa) promoter regions in woody stem tissues.the increasing focus on plantation forestry as a renewable source of cellulosic biomass has emphasized the need for tools to study the unique biology of woody genera such as eucalyptus, populus and pinus. the domestication of these woody crops is hampered by long generation times, and breeders are now looking to molecular approaches such as marker-assisted breeding and genetic modification to accelerate tree improvement. much of what is known about genes involved in the growth and development of ...201323132521
effects of pretreatment on morphology, chemical composition and enzymatic digestibility of eucalyptus bark: a potentially valuable source of fermentable sugars for biofuel production - part 1.in recent years, the growing demand for biofuels has encouraged the search for different sources of underutilized lignocellulosic feedstocks that are available in sufficient abundance to be used for sustainable biofuel production. much attention has been focused on biomass from grass. however, large amounts of timber residues such as eucalyptus bark are available and represent a potential source for conversion to bioethanol. in the present paper, we investigate the effects of a delignification p ...201323657132
[molecular evolution of orf1 gene in plant ltr-retrotransposons].ltr-retrotransposons are the major dna components in plant genomes. they usually contain gag and pol, two genes necessary for transpositinal process. our previous study on soybean genome annotation identified a sare ltr-retrotransposon family, which carries the third gene, orf1. using a bioinformatics approach, we here reported that 7 out of 33 sequenced genomes have some ltr-retrotransposons with an extra orf1 gene/gene fragment (approximately 1-2 kb) in the region between 5' ltr and gag gene, ...201324400486
ion torrent pgm as tool for fungal community analysis: a case study of endophytes in eucalyptus grandis reveals high taxonomic diversity.the kingdom fungi adds substantially to the diversity of life, but due to their cryptic morphology and lifestyle, tremendous diversity, paucity of formally described specimens, and the difficulty in isolating environmental strains into culture, fungal communities are difficult to characterize. this is especially true for endophytic communities of fungi living in healthy plant tissue. the developments in next generation sequencing technologies are, however, starting to reveal the true extent of f ...201324358124
the tonoplast intrinsic aquaporin (tip) subfamily of eucalyptus grandis: characterization of egtip2, a root-specific and osmotic stress-responsive gene.aquaporins have important roles in various physiological processes in plants, including growth, development and adaptation to stress. in this study, a gene encoding a root-specific tonoplast intrinsic aquaporin (tip) from eucalyptus grandis (named egtip2) was investigated. the root-specific expression of egtip2 was validated over a panel of five eucalyptus organ/tissues. in eucalyptus roots, egtip2 expression was significantly induced by osmotic stress imposed by peg treatment. histochemical ana ...201324157213
ultra-high performance liquid chromatography coupled to mass spectrometry applied to the identification of valuable phenolic compounds from eucalyptus wood.ultra-high performance liquid chromatography (uhplc) was applied for the first time in the analysis of wood extracts. the potential of this technique coupled to ion trap mass spectrometry in the rapid and effective detection and identification of bioactive components in complex vegetal samples was demonstrated. several dozens of compounds were detected in less than 30min of analysis time, corresponding to more than 3-fold reduction in time, when compared to conventional hplc analysis of similar ...201324055752
assessment of cryptococcus albidus for biopulping of eucalyptus.cryptococcus albidus shows delignification activity in nature. it was used for the biopulping of eucalyptus wood (eucalyptus grandis) to access its potential for industrial application in the pulp and paper industry. enzyme analysis on days 15, 30, and 60 showed the presence of laccase and xylanase as key enzymes. the production of endo-glucanase (cmcase) and exo-glucanase (fpase) was very low. scanning electron microscopy (sem) showed the surface colonization of wood and loosening of wood fiber ...201323876135
dynamics of soil exploration by fine roots down to a depth of 10 m throughout the entire rotation in eucalyptus grandis plantations.although highly weathered soils cover considerable areas in tropical regions, little is known about exploration by roots in deep soil layers. intensively managed eucalyptus plantations are simple forest ecosystems that can provide an insight into the belowground growth strategy of fast-growing tropical trees. fast exploration of deep soil layers by eucalypt fine roots may contribute to achieving a gross primary production that is among the highest in the world for forests. soil exploration by fi ...201323847645
impact of decomposing cinnamomum septentrionale leaf litter on the growth of eucalyptus grandis saplings.a pot experiment was performed to study the impact of decomposing cinnamomum septentrionale leaf litter on the growth of eucalyptus grandis saplings. the experimental design scheme was 0 (ck), 40 (a1), 80 (a2) and 120 g pot(-1) (a3) of e. grandis leaves, and changes in the volatile oil chemical composition during litter decomposition were assessed in the present study. the results showed that c. septentrionale leaf litter inhibited the growth of e. grandis saplings, as determined by the height, ...201323835358
the identification and differential expression of eucalyptus grandis pathogenesis-related genes in response to salicylic acid and methyl jasmonate.two important role players in plant defence response are the phytohormones salicylic acid (sa) and jasmonic acid (ja); both of which have been well described in model species such as arabidopsis thaliana. several pathogenesis related (pr) genes have previously been used as indicators of the onset of sa and ja signaling in arabidopsis. this information is lacking in tree genera such as eucalyptus. the aim of this study was to characterize the transcriptional response of pr genes (egrpr2, egrpr3, ...201323508356
[effects of eucalyptus grandis leaf litter at its early stage of decomposition on the growth and photosynthetic characteristics of cichorium intybus].from march to may, 2010, a pot experiment was conducted to investigate the effects of eucalyptus grandis leaf litter at its early stage of decomposition on the growth and photosynthetic characteristics of cichorium intybus. four treatments with different application rate of the leaf litter, i.e., 0 g x pot(-1) (ck), 30 g x pot(-1) (a1), 60 g x pot(-1) (a2), and 90 g x pot(-1) (a3), were installed. each pot contained 12 kg soil mixed with the leaf litter, and then, c. intybus was sown. the growth ...201324175509
mixing eucalyptus and acacia trees leads to fine root over-yielding and vertical segregation between species.the consequences of diversity on belowground processes are still poorly known in tropical forests. the distributions of very fine roots (diameter <1 mm) and fine roots (diameter <3 mm) were studied in a randomized block design close to the harvest age of fast-growing plantations. a replacement series was set up in brazil with mono-specific eucalyptus grandis (100e) and acacia mangium (100a) stands and a mixture with the same stocking density and 50% of each species (50a:50e). the total fine root ...201323180423
uncovering the defence responses of eucalyptus to pests and pathogens in the genomics age.long-lived tree species are subject to attack by various pests and pathogens during their lifetime. this problem is exacerbated by climate change, which may increase the host range for pathogens and extend the period of infestation by pests. plant defences may involve preformed barriers or induced resistance mechanisms based on recognition of the invader, complex signalling cascades, hormone signalling, activation of transcription factors and production of pathogenesis-related (pr) proteins with ...201425261123
wgdb: wood gene database with search interface.wood quality can be defined in terms of particular end use with the involvement of several traits. over the last fifteen years researchers have assessed the wood quality traits in forest trees. the wood quality was categorized as: cell wall biochemical traits, fibre properties include the microfibril angle, density and stiffness in loblolly pine [1]. the user friendly and an open-access database has been developed named wood gene database (wgdb) for describing the wood genes along the informatio ...201424516325
photosynthetic and anatomical responses of eucalyptus grandis leaves to potassium and sodium supply in a field experiment.although vast areas in tropical regions have weathered soils with low potassium (k) levels, little is known about the effects of k supply on the photosynthetic physiology of trees. this study assessed the effects of k and sodium (na) supply on the diffusional and biochemical limitations to photosynthesis in eucalyptus grandis leaves. a field experiment comparing treatments receiving k (+k) or na (+na) with a control treatment (c) was set up in a k-deficient soil. the net co2 assimilation rates w ...201423663049
in vitro scleroderma laeve and eucalyptus grandis mycorrhization and analysis of atp6, 17s rdna, and ras gene expression during ectomycorrhizal formation.the interaction between fungi and plants that form ectomycorrhizae (ecm) promotes alterations in the gene expression profiles of both organisms. fungal genes expression related to metabolism were evaluated at the pre-symbiotic stage and during the ecm development between scleroderma laeve and eucalyptus grandis. partial sequences of atp synthase (atp6), translation elongation factor (ef1α), the ras protein (ras), and the 17s rdna genes were isolated. the expression of the atp6 and 17s rdna genes ...201425080195
profiling micrornas in eucalyptus grandis reveals no mutual relationship between alterations in mir156 and mir172 expression and adventitious root induction during development.the change from juvenile to mature phase in woody plants is often accompanied by a gradual loss of rooting ability, as well as by reduced microrna (mir) 156 and increased mir172 expression.201424965948
flavonoid supplementation affects the expression of genes involved in cell wall formation and lignification metabolism and increases sugar content and saccharification in the fast-growing eucalyptus hybrid e. urophylla x e. grandis.eucalyptus species are the most widely planted hardwood species in the world and are renowned for their rapid growth and adaptability. in brazil, one of the most widely grown eucalyptus cultivars is the fast-growing eucalyptus urophylla x eucalyptus grandis hybrid. in a previous study, we described a chemical characterization of these hybrids when subjected to flavonoid supplementation on 2 distinct timetables, and our results revealed marked differences between the wood composition of the treat ...201425407319
streptomyces rhizobacteria modulate the secondary metabolism of eucalyptus plants.the genus eucalyptus comprises economically important species, such as eucalyptus grandis and eucalyptus globulus, used especially as a raw material in many industrial sectors. species of eucalyptus are very susceptible to pathogens, mainly fungi, which leads to mortality of plant cuttings in rooting phase. one alternative to promote plant health and development is the potential use of microorganisms that act as agents for biological control, such as plant growth-promoting rhizobacteria (pgpr). ...201425394796
eucalyptus grandis hill ex maiden inoculated with pisolithus microcarpus (ufsc-pt116) in land subject to the sandy process in southern brazil.eucalypts is one of the main species used for commercial reforestation in the rio grande do sul state, brazil. this study aimed to evaluate the survival and early growth of eucalyptus trees in an area subject to sandy process after three years of growth. the eucalyptus grandis seedlings were grown in a greenhouse, inoculated or not with the isolated ectomycorrhizal pisolithus microcarpus (ufsc-pt116), produced in peat or entisol. after 120 days, the seedlings were transplanted to an area subject ...201425763017
phosphorus limits eucalyptus grandis seedling growth in an unburnt rain forest soil.although rain forest is characterized as pyrophobic, pyrophilic giant eucalypts grow as rain forest emergents in both temperate and tropical australia. in temperate australia, such eucalypts depend on extensive, infrequent fires to produce conditions suitable for seedling growth. little is known, however, about constraints on seedlings of tropical giant eucalypts. we tested whether seedlings of eucalyptus grandis experience edaphic constraints similar to their temperate counterparts. we hypothes ...201425339968
eucalyptus tolerance mechanisms to lanthanum and cerium: subcellular distribution, antioxidant system and thiol pools.guanglin 9 (eucalyptus grandis × eucalyptus urophlla) and eucalyptus grandis 5 are two eucalyptus species which have been found to grow normally in soils contaminated with lanthanum and cerium, but the tolerance mechanisms are not clear yet. in this study, a pot experiment was conducted to investigate the tolerance mechanisms of the eucalyptus to lanthanum and cerium. cell walls stored 45.40-63.44% of the metals under lanthanum or cerium stress. peroxidase and catalase activities enhanced with i ...201425303462
genome-wide characterization and expression profiling of the auxin response factor (arf) gene family in eucalyptus grandis.auxin is a central hormone involved in a wide range of developmental processes including the specification of vascular stem cells. auxin response factors (arf) are important actors of the auxin signalling pathway, regulating the transcription of auxin-responsive genes through direct binding to their promoters. the recent availability of the eucalyptus grandis genome sequence allowed us to examine the characteristics and evolutionary history of this gene family in a woody plant of high economic i ...201425269088
gene expression profiling in juvenile and mature cuttings of eucalyptus grandis reveals the importance of microtubule remodeling during adventitious root formation.the ability to form adventitious roots (ar) is an economically important trait that is lost during the juvenile-to-mature phase change in woody plants. auxin treatment, which generally promotes rooting in juvenile cuttings, is often ineffective when applied to mature cuttings. the molecular basis for this phenomenon in eucalyptus grandis was addressed here.201425266376
the genome of eucalyptus grandis.eucalypts are the world's most widely planted hardwood trees. their outstanding diversity, adaptability and growth have made them a global renewable resource of fibre and energy. we sequenced and assembled >94% of the 640-megabase genome of eucalyptus grandis. of 36,376 predicted protein-coding genes, 34% occur in tandem duplications, the largest proportion thus far in plant genomes. eucalyptus also shows the highest diversity of genes for specialized metabolites such as terpenes that act as che ...201424919147
cold pretreatment amplifies the responses of in vitro eucalyptus grandis shoots to cryopreparative drying.eucalyptus grandis is an important commercial forestry species in south africa. little is known about its response (which is both drought and cold sensitive) to cold exposure and subsequent drying.201424872158
formation of carbonyl groups on cellulose during ozone treatment of pulp: consequences for pulp bleaching.the formation of carbonyl groups during the ozone treatment (z) of eucalyptus (eucalyptus grandis and eucalyptus urophylla hybrid) kraft pulps and their behaviors during subsequent alkaline stages were investigated by the ccoa method with carbazole-9-carboxylic acid [2-(2-aminooxethoxy)-ethoxy] amide (ccoa) as the carbonyl-selective fluorescence label. several pulp samples with or without lignin and hexenuronic acids (hexa) were used to elucidate the effects of these components when present in u ...201424815405
effects of potassium and sodium supply on drought-adaptive mechanisms in eucalyptus grandis plantations.a basic understanding of nutrition effects on the mechanisms involved in tree response to drought is essential under a future drier climate. a large-scale throughfall exclusion experiment was set up in brazil to gain an insight into the effects of potassium (k) and sodium (na) nutrition on tree structural and physiological adjustments to water deficit. regardless of the water supply, k and na supply greatly increased growth and leaf area index (lai) of eucalyptus grandis trees over the first 3 y ...201424725318
programmed cell death and necrosis during cryopreparative drying of in vitro eucalyptus grandis axillary buds.in preparation for cryopreservation, eucalyptus grandis in vitro axillary buds were dried over silica gel. pretreatment of the buds with 5 mg per l aba resulted in partial resistance to water loss (0.76 to 0.45 g per g fresh mass basis) as compared with untreated buds (0.76 to 0.33 g per g) and was associated with the retention of viability (70 vs. 55%). the loss of viability of the dried buds was protracted over several days. ultrastructural examination and vital staining demonstrated cellular ...201424441369
measured and modeled interactive effects of potassium deficiency and water deficit on gross primary productivity and light-use efficiency in eucalyptus grandis plantations.global climate change is expected to increase the length of drought periods in many tropical regions. although large amounts of potassium (k) are applied in tropical crops and planted forests, little is known about the interaction between k nutrition and water deficit on the physiological mechanisms governing plant growth. a process-based model (maespa) parameterized in a split-plot experiment in brazil was used to gain insight into the combined effects of k deficiency and water deficit on absor ...201525430918
the capacity to cope with climate warming declines from temperate to tropical latitudes in two widely distributed eucalyptus species.as rapid climate warming creates a mismatch between forest trees and their home environment, the ability of trees to cope with warming depends on their capacity to physiologically adjust to higher temperatures. in widespread species, individual trees in cooler home climates are hypothesized to more successfully acclimate to warming than their counterparts in warmer climates that may approach thermal limits. we tested this prediction with a climate-shift experiment in widely distributed eucalyptu ...201525378195
genome-wide analysis of the lignin toolbox of eucalyptus grandis.lignin, a major component of secondary cell walls, hinders the optimal processing of wood for industrial uses. the recent availability of the eucalyptus grandis genome sequence allows comprehensive analysis of the genes encoding the 11 protein families specific to the lignin branch of the phenylpropanoid pathway and identification of those mainly involved in xylem developmental lignification. we performed genome-wide identification of putative members of the lignin gene families, followed by com ...201525684249
the fasciclin-like arabinogalactan protein family of eucalyptus grandis contains members that impact wood biology and biomechanics.fasciclin-like arabinogalactan protein (fla) families have been identified and characterised in key plant species, with some members exhibiting functional specialization. here we identify the fla family of eucalyptus grandis, and investigate the roles of three single-fas domain flas, with particular focus on secondary cell-wall formation and wood properties. we use various in-silico approaches to identify and characterise e. grandis genome flas, and perform phylogenetic comparisons with other sp ...201525676073
comparative interrogation of the developing xylem transcriptomes of two wood-forming species: populus trichocarpa and eucalyptus grandis.wood formation is a complex developmental process governed by genetic and environmental stimuli. populus and eucalyptus are fast-growing, high-yielding tree genera that represent ecologically and economically important species suitable for generating significant lignocellulosic biomass. comparative analysis of the developing xylem and leaf transcriptomes of populus trichocarpa and eucalyptus grandis together with phylogenetic analyses identified clusters of homologous genes preferentially expres ...201525659405
the effect of elevated carbon dioxide on the interaction between eucalyptus grandis and diverse isolates of pisolithus sp. is associated with a complex shift in the root transcriptome.using the newly available genome for eucalyptus grandis, we sought to determine the genome-wide traits that enable this host to form mutualistic interactions with ectomycorrhizal (ecm) pisolithus sp. and to determine how future predicted concentrations of atmospheric carbon dioxide (co2 ) will affect this relationship. we analyzed the physiological and transcriptomic responses of e. grandis during colonization by different pisolithus sp. isolates under conditions of ambient (400 ppm) and elevate ...201525377589
integrated hot-compressed water and laccase-mediator treatments of eucalyptus grandis fibers: structural changes of fiber and lignin.eucalyptus grandis fibers were treated with hot-compressed water (hcw) and laccase mediator to enhance the fiber characteristics and to produce an active lignin substrate for binderless fiberboard production. the composition, morphology, and crystallinity index (cri) analysis of fibers showed that the hcw treatment increased the cri and lignin content of the treated fibers through partial removal of hemicelluloses. simultaneously, the hcw treatment produced some granules and holes on the surface ...201525639522
eucalyptus urograndis stem proteome is responsive to short-term cold stress.eucalyptus urograndis is a hybrid eucalyptus of major economic importance to the brazilian pulp and paper industry. although widely used in forest nurseries around the country, little is known about the biochemical changes imposed by environmental stress in this species. in this study, we evaluated the changes in the stem proteome after short-term stimulation by exposure to low temperature. using two-dimensional gel electrophoresis coupled to high-resolution mass spectrometry-based protein ident ...201526273222
transcriptionally active ltr retrotransposons in eucalyptus genus are differentially expressed and insertionally polymorphic.in eucalyptus genus, studies on genome composition and transposable elements (tes) are particularly scarce. nearly half of the recently released eucalyptus grandis genome is composed by retrotransposons and this data provides an important opportunity to understand te dynamics in eucalyptus genome and transcriptome.201526268941
modifying solubility of polymeric xylan extracted from eucalyptus grandis and sugarcane bagasse by suitable side chain removing enzymes.α-l-arabinofuranosidase (abfb) and novel α-d-glucuronidase (agu1b) enzymes were applied for selective hydrolysis of beechwood (fagus sylvatica) xylan (sigma-aldrich) as well as xylans extracted from eucalyptus grandis and sugarcane (saccharum officinarum l.) bagasse, leading to precipitation of these carbohydrate biopolymers. hemicellulose extraction was performed with two mild-alkali methods, höije and pinto. precipitation occurred after removal of 67, 40 and 16% 4-o-methyl-d-glucuronic acid (m ...201526256174
structural, evolutionary and functional analysis of the nac domain protein family in eucalyptus.nac domain transcription factors regulate many developmental processes and stress responses in plants and vary widely in number and family structure. we analysed the characteristics and evolution of the nac gene family of eucalyptus grandis, a fast-growing forest tree in the rosid order myrtales. nac domain genes identified in the e. grandis genome were subjected to amino acid sequence, phylogenetic and motif analyses. transcript abundance in developing tissues and abiotic stress conditions in e ...201525385212
high-resolution genetic maps of eucalyptus improve eucalyptus grandis genome assembly.genetic maps are key tools in genetic research as they constitute the framework for many applications, such as quantitative trait locus analysis, and support the assembly of genome sequences. the resequencing of the two parents of a cross between eucalyptus urophylla and eucalyptus grandis was used to design a single nucleotide polymorphism (snp) array of 6000 markers evenly distributed along the e. grandis genome. the genotyping of 1025 offspring enabled the construction of two high-resolution ...201525385325
genome-wide patterns of recombination, linkage disequilibrium and nucleotide diversity from pooled resequencing and single nucleotide polymorphism genotyping unlock the evolutionary history of eucalyptus grandis.we used high-density single nucleotide polymorphism (snp) data and whole-genome pooled resequencing to examine the landscape of population recombination (ρ) and nucleotide diversity (θw ), assess the extent of linkage disequilibrium (r(2) ) and build the highest density linkage maps for eucalyptus. at the genome-wide level, linkage disequilibrium (ld) decayed within c. 4-6 kb, slower than previously reported from candidate gene studies, but showing considerable variation from absence to complete ...201526079595
the transcriptome and terpene profile of eucalyptus grandis reveals mechanisms of defense against the insect pest, leptocybe invasa.plants have evolved complex defenses that allow them to protect themselves against pests and pathogens. however, there is relatively little information regarding the eucalyptus defensome. leptocybe invasa is one of the most damaging pests in global eucalyptus forestry, and essentially nothing is known regarding the molecular mechanisms governing the interaction between the pest and host. the aim of the study was to investigate changes in the transcriptional landscape and terpene profile of a res ...201525948810
transcriptome and hormone profiling reveals eucalyptus grandis defence responses against chrysoporthe austroafricana.eucalyptus species and interspecific hybrids exhibit valuable growth and wood properties that make them a highly desirable commodity. however, these trees are challenged by a wide array of biotic stresses during their lifetimes. the eucalyptus grandis reference genome sequence provides a resource to study pest and pathogen defence mechanisms in long-lived woody plants. e. grandis trees are generally susceptible to chrysoporthe austroafricana, a causal agent of stem cankers on eucalypts. the aim ...201525903559
comprehensive genome-wide analysis of the aux/iaa gene family in eucalyptus: evidence for the role of egriaa4 in wood formation.auxin plays a pivotal role in various plant growth and development processes, including vascular differentiation. the modulation of auxin responsiveness through the auxin perception and signaling machinery is believed to be a major regulatory mechanism controlling cambium activity and wood formation. to gain more insights into the roles of key aux/iaa gene regulators of the auxin response in these processes, we identified and characterized members of the aux/iaa family in the genome of eucalyptu ...201525577568
protein domain evolution is associated with reproductive diversification and adaptive radiation in the genus eucalyptus.eucalyptus is a pivotal genus within the rosid order myrtales with distinct geographic history and adaptations. comparative analysis of protein domain evolution in the newly sequenced eucalyptus grandis genome and other rosid lineages sheds light on the adaptive mechanisms integral to the success of this genus of woody perennials. we reconstructed the ancestral domain content to elucidate the gain, loss and expansion of protein domains and domain arrangements in eucalyptus in the context of rosi ...201525494981
comparative analysis of plant carbohydrate active enzymes and their role in xylogenesis.carbohydrate metabolism is a key feature of vascular plant architecture, and is of particular importance in large woody species, where lignocellulosic biomass is responsible for bearing the bulk of the stem and crown. since carbohydrate active enzymes (cazymes) in plants are responsible for the synthesis, modification and degradation of carbohydrate biopolymers, the differences in gene copy number and regulation between woody and herbaceous species have been highlighted previously. there are sti ...201525994181
genome-wide discovery and validation of eucalyptus small rnas reveals variable patterns of conservation and diversity across species of myrtaceae.micro rnas are a class of small non coding rnas of 20-24 nucleotides transcribed as single stranded precursors from mir gene loci. initially described as post-transcriptional regulators involved in development, two decades ago, mirnas have been proven to regulate a wide range of processes in plants such as germination, morphology and responses to biotic and abiotic stress. despite wide conservation in plants, a number of mirnas are lineage specific. we describe the first genome wide survey of eu ...201526714854
a newly identified passive hyperaccumulator eucalyptus grandis × e. urophylla under manganese stress.manganese (mn) is an essential micronutrient needed for plant growth and development, but can be toxic to plants in excess amounts. however, some plant species have detoxification mechanisms that allow them to accumulate mn to levels that are normally toxic, a phenomenon known as hyperaccumulation. these species are excellent candidates for developing a cost-effective remediation strategy for mn-polluted soils. in this study, we identified a new passive mn-hyperaccumulator eucalyptus grandis × e ...201526327118
investigating the molecular underpinnings underlying morphology and changes in carbon partitioning during tension wood formation in eucalyptus.tension wood has distinct physical and chemical properties, including altered fibre properties, cell wall composition and ultrastructure. it serves as a good system for investigating the genetic regulation of secondary cell wall biosynthesis and wood formation. the reference genome sequence for eucalyptus grandis allows investigation of the global transcriptional reprogramming that accompanies tension wood formation in this global wood fibre crop. we report the first comprehensive analysis of ph ...201525388807
a novel genome-wide microsatellite resource for species of eucalyptus with linkage-to-physical correspondence on the reference genome sequence.keystone species in their native ranges, eucalypts, are ecologically and genetically very diverse, growing naturally along extensive latitudinal and altitudinal ranges and variable environments. besides their ecological importance, eucalypts are also the most widely planted trees for sustainable forestry in the world. we report the development of a novel collection of 535 microsatellites for species of eucalyptus, 494 designed from ests and 41 from genomic libraries. a selected subset of 223 was ...201525146326
genome-wide analysis of the ap2/erf family in eucalyptus grandis: an intriguing over-representation of stress-responsive dreb1/cbf genes.the ap2/erf family includes a large number of developmentally and physiologically important transcription factors sharing an ap2 dna-binding domain. among them dreb1/cbf and dreb2 factors are known as master regulators respectively of cold and heat/osmotic stress responses.201525849589
explosive tandem and segmental duplications of multigenic families in eucalyptus grandis.plant organisms contain a large number of genes belonging to numerous multigenic families whose evolution size reflects some functional constraints. sequences from eight multigenic families, involved in biotic and abiotic responses, have been analyzed in eucalyptus grandis and compared with arabidopsis thaliana. two transcription factor families apetala 2 (ap2)/ethylene responsive factor and gras, two auxin transporter families pin-formed and aux/lax, two oxidoreductase families (ascorbate perox ...201525769696
the floral transcriptome of eucalyptus grandis.as a step toward functional annotation of genes required for floral initiation and development within the eucalyptus genome, we used short read sequencing to analyze transcriptomes of floral buds from early and late developmental stages, and compared these with transcriptomes of diverse vegetative tissues, including leaves, roots, and stems. a subset of 4807 genes (13% of protein-coding genes) were differentially expressed between floral buds of either stage and vegetative tissues. a similar pro ...201525353719
does diatomaceous earth control leaf-cutter ants (hymenoptera: formicidae) in the eucalyptus plantations?genus atta includes some of the most important formicidae leaf cutter ants which cause extensive damage to the eucalyptus plantations. atta sexdens rubropilosa forel, one of the chief pests in brazilian reforestation, can restrict and reduce forest productivity by its intense and constant leaf-cutting activities on plants at all stages. therefore, the demand for new products to control a. sexdens rubropilosa indicates the study of the utilization of the dry powder formulation of diatomaceous ear ...201526470237
genome-wide mapping of histone h3 lysine 4 trimethylation in eucalyptus grandis developing xylem.histone modifications play an integral role in plant development, but have been poorly studied in woody plants. investigating chromatin organization in wood-forming tissue and its role in regulating gene expression allows us to understand the mechanisms underlying cellular differentiation during xylogenesis (wood formation) and identify novel functional regions in plant genomes. however, woody tissue poses unique challenges for using high-throughput chromatin immunoprecipitation (chip) technique ...201525957781
temporal dynamics of the response to al stress in eucalyptus grandis × eucalyptus camaldulensis.lipid peroxidation and root elongation of eucalyptus grandis × eucalyptus camaldulensis were studied under stress conditions in response to aluminum (al), a metal known to limit agricultural productivity in acidic soils primarily due to reduced root elongation. in brazil, the grancam 1277 hybrid (e. grandis × e. camaldulensis) has been planted in the "cerrado", a region of the country with a wide occurrence of acidic soils. the present study demonstrated that the hybrid exhibited root growth red ...201526062119
the eucalyptus terpene synthase gene family.terpenoids are abundant in the foliage of eucalyptus, providing the characteristic smell as well as being valuable economically and influencing ecological interactions. quantitative and qualitative inter- and intra- specific variation of terpenes is common in eucalypts.201526062733
hylax bahiensis bechyné (chrysomelidae: eumolpinae): a new potential pest of eucalyptus and species used for atlantic rainforest restoration.hylax bahiensis bechyné (coleoptera: chrysomelidae), a new pest of forest species, including eucalyptus (hybrid eucalyptus urophylla x eucalyptus grandis), joannesia princeps, mimosa artemisiana, croton urucurana, croton floribundus, and senna multijuga is recorded. the insect attack in clonal eucalyptus plantations and in forest restoration areas between 2010 and 2013 in the states of espírito santo, bahia and minas gerais, brasil, was observed for the first time. the outbreaks generally occurr ...201526013018
the eucalyptus grandis nbs-lrr gene family: physical clustering and expression hotspots.eucalyptus grandis is a commercially important hardwood species and is known to be susceptible to a number of pests and pathogens. determining mechanisms of defense is therefore a research priority. the published genome for e. grandis has aided the identification of one important class of resistance (r) genes that incorporate nucleotide binding sites and leucine-rich repeat domains (nbs-lrr). using an iterative search process we identified nbs-lrr gene models within the e. grandis genome. we cha ...201526793216
the eucalyptus grandis r2r3-myb transcription factor family: evidence for woody growth-related evolution and function.the r2r3-myb family, one of the largest transcription factor families in higher plants, controls a wide variety of plant-specific processes including, notably, phenylpropanoid metabolism and secondary cell wall formation. we performed a genome-wide analysis of this superfamily in eucalyptus, one of the most planted hardwood trees world-wide. a total of 141 predicted r2r3-myb sequences identified in the eucalyptus grandis genome sequence were subjected to comparative phylogenetic analyses with ar ...201525250741
behavioral response of aedes aegypti (diptera: culicidae) larvae to synthetic and natural attractants and repellents.aedes aegypti (l.) (diptera: culicidae) is the key vector of three important arboviral diseases: dengue, yellow fever, and chikungunya. immature stages of this species inhabit human-made containers placed in residential landscapes. in this study, we evaluated a few compounds in a sensitive behavioral assay with ae. aegypti larvae. the orientation of larvae to different compounds was surveyed using a performance index (pi). the pi represents the response to each odorant, where a value of +1 is in ...201526352935
localization and transcriptional responses of chrysoporthe austroafricana in eucalyptus grandis identify putative pathogenicity factors.chrysoporthe austroafricana is a fungal pathogen that causes the development of stem cankers on susceptible eucalyptus grandis trees. clones of e. grandis that are partially resistant and highly susceptible have been identified based on the extent of lesion formation on the stem upon inoculation with c. austroafricana. these interactions have been used as a model pathosystem to enhance our understanding of interactions between pathogenic fungi and woody hosts, which may be different to herbaceou ...201628008326
effects of an inducible aiia gene on disease resistance in eucalyptus urophylla × eucalyptus grandis.n-acyl-homoserine lactones (ahls) are metabolites of mostly gram-negative bacteria and are critical signaling molecules in bacterial quorum-sensing systems. at threshold concentrations, ahls can activate the expression of pathogenic genes and induce diseases. therefore, reducing ahl concentrations is a key point of disease control in plants. ahl-lactonase, which is expressed by aiia, is widespread in bacillus sp and can hydrolyze ahls. in the present study, we cloned aiia from bacillus subtilis ...201626905275
defense responses in plants of eucalyptus elicited by streptomyces and challenged with botrytis cinerea.elicitation of e. grandis plants with streptomyces pm9 reduced the gray-mold disease, through increasing the levels of enzymes directly related to the induction of plant defense responses, and accumulation of specific phenolic compounds. members of eucalyptus are economically important woody species, especially as a raw material in many industrial sectors. species of this genus are susceptible to pathogens such as botrytis cinerea (gray mold). biological control of plant diseases using rhizobact ...201626794965
eucalyptus hairy roots, a fast, efficient and versatile tool to explore function and expression of genes involved in wood formation.eucalyptus are of tremendous economic importance being the most planted hardwoods worldwide for pulp and paper, timber and bioenergy. the recent release of the eucalyptus grandis genome sequence pointed out many new candidate genes potentially involved in secondary growth, wood formation or lineage-specific biosynthetic pathways. their functional characterization is, however, hindered by the tedious, time-consuming and inefficient transformation systems available hitherto for eucalypts. to overc ...201626579999
ft overexpression induces precocious flowering and normal reproductive development in eucalyptus.eucalyptus trees are among the most important species for industrial forestry worldwide. however, as with most forest trees, flowering does not begin for one to several years after planting which can limit the rate of conventional and molecular breeding. to speed flowering, we transformed a eucalyptus grandis × urophylla hybrid (sp7) with a variety of constructs that enable overexpression of flowering locus t (ft). we found that ft expression led to very early flowering, with events showing flor ...201626132805
intraspecific competition between ectomycorrhizal pisolithus microcarpus isolates impacts plant and fungal performance under elevated co2 and temperature.root systems are simultaneously colonized by multiple individuals of mycorrhizal fungi. intraspecific competitive interactions between fungal isolates are likely to affect both fungal and plant performance and be influenced by abiotic factors. here, we assessed the impact of intraspecific competition between three pisolithus microcarpus isolates on the establishment of, and benefit derived from, symbioses with eucalyptus grandis seedlings. we investigated the outcomes of competition under ambien ...201627222224
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