Eucalyptus Species Research Articles

Identification of cold stress-responsive microRNAs and mRNA in Eucalyptus grandis

MicroRNAs (miRNAs) are a class of non-coding endogenous small RNAs that play a crucial role in plant growth, development, morphogenesis, signal transduction, and stress response. miRNAs negatively regulate gene expression at post-transcriptional level, causing translational repression or/and transcript degradation. The role of miRNAs in understanding gene expression reprogramming under different stress types has been pointed out. Low temperature is an important factor limiting the growth and distribution of plants. Under cold stress, plants attempt to maintain homeostasis and acquire cold acclimation, which involves extensive gene expression and metabolism reprogramming. Particularly, cold stress limits the distribution and productivity of Eucalyptus plantations, significantly affecting the survival and yield of seedlings. Despite the consensus on the relevance of miRNAs in regulating gene expression in plants and Eucalyptus's huge importance for the forest industry worldwide, information on miRNAs in Eucalyptus is still very scarce. In this work, we studied the miRNA and mRNA response to cold stress in E. grandis, one of Uruguay's most widely planted Eucalyptus species. Since seedlings are more susceptible to chilling injury, we subjected 3 different genotypes of seedlings to a cold shock followed by a 12-hour recovery phase at 25°C, analyzing a total of 4 treatments (G1: control, G2: 12 hours at 4°C, G3: 24 hours at 4°C, and G4: 24 hours at 4°C + 12 hours at 25°C). Cold treatment was evaluated measuring the electrolyte leakage. The membrane's damage was verified at 12 hours at 4°C, increasing the parameter at 24 hours. The membrane was partially recovered after 12 hours at 25°C, decreasing the electrolyte leakage. Transcriptome analysis showed an increase in the expression of genes belonging to the AP2|ERF family, including the DREB1|CBF genes —a group of genes that control the activation of protection mechanisms under cold stress. Within down-regulated genes, GO terms related to photosynthesis were enriched. Several conserved and new microRNAs were identified, but only a minority changed their expression with cold treatment, among them, the miR828. This microRNA was down-regulated after the cold treatment, which, according to several studies, induces the expression of anthocyanins. Anthocyanin levels are known to increase under different conditions, including cold stress. These results provided data on gene silencing mechanisms in E. grandis and allowed characterizing the response to cold mediated by small RNAs. This data could be potentially used by selection and improvement programs seeking to select clones more tolerant to cold stress.

Feasibility of using Uruguayan Eucalyptus for sustainable timber connections

Uruguayan fast-growing Eucalyptus grandis has emerged as a promising sustainable material for structural building components. However, traditional methods for joining timber components, such as beams and columns, typically rely on metallic fasteners, and mass timber products like glued laminated timber often use adhesives. In response to growing environmental concerns over the past five years, researchers have explored the substitution of petroleum-based adhesives and metallic fasteners with wooden pegs or dowels. To create effective connections, these dowels must be fashioned from denser wood than the lamellae. The embedment strength of both woods with varying densities is a critical factor in the design of timber connections using fasteners. This paper presents the results of a structural performance analysis of fully eucalyptus dowelled connections, constructed by combining Eucalyptus grandis (? = 473.2 kg|m3, COV = 13.2 %) for the lamellae and Eucalyptus tereticornis (? = 1042.4 kg|m^3, COV = 3.2 %) for the dowel. The bending strength of dowels was determined for specimens with 15- and 18-mm diameters, yielding 173.0 MPa (COV = 10.7 %) and 184.0 MPa (COV = 8.3 %), respectively. Four series of connections were tested, varying the fiber orientation of the lamellae (longitudinal L, transversal T) and dowel diameter (15, 18), following the half-hole method described in ASTM D 5764 standard. The embedment strength values obtained for the connections are as follows: L-15 = 31.3 MPa (COV = 9.3 %), T-15 = 11.2 MPa (COV = 28.7 %), L-18 = 29.6 MPa (COV = 10.1 %), T-18 = 16.7 MPa (COV = 25.7 %). These results reveal that the longitudinal application of the load, aligned with the fiber orientation, exhibits approximately twice the strength of the transversal orientation, with less variability in the outcomes. Furthermore, a strong correlation with the lamellae density was observed, while no significant dependency on the dowel diameter was found. These findings represent a crucial initial step in characterizing and designing glue-free wood-wood connections. The utilization of different eucalyptus species presents a promising path for the development of connections that promote more sustainable timber construction systems.

Drought effects on water status, plant hydraulics and growth in Eucalyptus grandis and clonal hybrids with red gums and E. urophylla

Afforestation with Eucalyptus species occupies over a million hectares in Uruguay, and E. grandis is one of the most cultivated species, mainly for the timber industry. However, the increasing frequency of drought and heat waves in the region raises the need to evaluate hybrids with red gums (E. camaldulensis and E. tereticornis), since they are known to sustain gas exchange and growth even under severe drought conditions. Hybrids with E. urophylla are also evaluated for plantations in subtropical regions. This study aimed to compare the effects of drought on plant water status, hydraulics, and growth of E. grandis (one clone; G), E. grandis × camaldulensis (one clone; GC), E. grandis × tereticornis (one clone; GT), and E. grandis × urophylla (two clones; GU1 and GU2). Two drought-stress cycles were applied to six-month rooted cuttings from mid-spring to early fall under greenhouse environmental conditions. Predawn leaf water potential (?pd), midday leaf water potential (?md), relative leaf water content (RWC), stomatal conductance (gs), and plant growth were measured across each cycle. Daily fluctuation of Ψ was calculated as ΔΨ = Ψpd - Ψmd. Specific (kS) and leaf-specific (kL) hydraulic conductivities and specific leaf area (SLA) were measured at the end of the experiment. Percentage loss of hydraulic conductivity (PLC) was calculated after removing the xylem embolism. Preliminary results suggest that both red-gum hybrids had higher PLC under moderate-to-severe drought, as a consequence of maintaining high water potential gradients to sustain transpiration and carbon fixation throughout the experiment. Under the well-watered condition, no clonal difference was observed. Also, GT showed a three-fold increase in cavitation and had 1.5 times stomatal opening (gs) than the most drought-sensitive clone (E. grandis, G), and exhibited the lowest reduction (17%) in diameter growth under drought. Also, GC attained the highest height at the end of the study, 40% higher than E. grandis, and increased by 25% the SLA under drought. GU clones reached the highest diameter at the end of the study. Correlations among functional variables are explored, and the water supply capacity to the foliage among clones is discussed for a better understanding of drought resistance strategies and growth performance of the different taxa.

Differential photosynthetic response to short- and long-term water deficit in Eucalyptus clones and hybrids with red gums

In Uruguay, there are more than a million hectares forested, and Eucalyptus species predominate. Drought events are becoming more frequent and affecting growth of the stands, especially during the early stages of the crop. Thus, the need arises to analyze the effects of such events on processes that determine growth, such as photosynthesis. The effect of water restriction on photosynthetic parameters was analyzed under greenhouse conditions in 6-months-old rooted cuttings of a clone of E. grandis (GG) and two hybrids with red gums: E. grandis × E. camaldulensis (GC) and E. grandis × E. tereticornis (GT). To define irrigation regimes water potential of substrate (MPa) was assessed using samples with increasing water content (%HR, w|w). Data was used to fit a water retention curve. Field’s capacity pot weight was determined on water drained pots. The control condition (WW) was established at Ψw = -0.15 MPa, while the water restricted (WR) was Ψw = -0.30 MPa. Treatments were maintained by manually water feeding for 16 weeks. Physiological parameters and photosynthetic pigments were recorded 6 and 16 weeks after water withholding to simulate short- and long-term water restriction scenarios, respectively. The following measurements were registered on current year fully expanded sun-exposed leaves: stomatal conductance (gs), maximum net assimilation rate (Amax), PSII quantum efficiency (FPSII), and proportion of chlorophyll and carotenoids in a HPLC elution (%Chl, %Carot). In the initial period, GG maintained Amax and gs between conditions. However, despite exhibiting lower gs in the restricted condition, GC maintained their Amax and GT increased in WR. There was no effect of treatment on photosynthetic pigments and energy partitioning varied little. At the end of the 16-week period, GG showed only decreased stomatal conductance values, and did not change photosynthetic pigment content nor Amax despite water restriction. Hybrids showed increasing values of FPSII, %Chl, and Amax. The PCA for long-term water restriction data showed that pigment content and FPSII may explain the increased assimilation rate of the hybrid clones under WR. Changes in water use efficiency due to higher % Chl in the restricted condition for both hybrids are discussed. There are few reports of increments in chlorophyll in Eucalyptus in drought. Recent information suggests that these types of strategies are adaptative responses that allow plants to cope with stress.

Macro- and Micronutrient Contents and Their Relationship with Growth in Six Eucalyptus Species

Knowing nutrient allocation dynamics in the tissues and the characteristics related to growth in different forest species is crucial to fertilization management and selecting better species for specific environments, ensuring greater fertilization efficiency and consequent sustainability in the forestry sector through the rational use of fertilizers. The objectives of this study were (i) to evaluate the content of macro- and micronutrients in different tissues of eucalyptus species and (ii) to relate them with their growth. The treatments were composed of six eucalyptus species (Eucalyptus camaldulensis Dehnh., Corymbia citriodora Hook., E. saligna Sm., E. grandis W. Hill ex Maiden, E. urograndis, and E. urophylla S. T. Blake). Macro- (nitrogen, phosphorus, potassium, calcium, magnesium, and sulfur) and micronutrient (boron, copper, iron, manganese, and zinc) contents were determined in the leaves, bark, and sapwood. To study the functional patterns in macro- and micronutrient contents, Canonical Variable Analysis (CVA) was performed. The first two canonical variables in nutrient content of leaves, bark, and sapwood and the growth variables of eucalyptus species accumulated values greater than 80% of variance. The species E. grandis and E. urograndis showed the highest means for volume and total height but showed no differences regarding the concentration of major elements in the tissues, except the iron content in the bark, which was higher compared to other species. CVA proved to be an excellent tool for understanding, identifying, and classifying the strategies of Eucalyptus sp. regarding the content of nutrients in the shoot biomass tissues and may support genetic improvement programs aiming at identifying potential species. Future research involving the use of remotely piloted aircraft and remote sensors could be a strategy to monitor nutrient contents in different parts of trees throughout the cycle of different eucalyptus species.

Ecophysiological adaptations shape distributions of closely related trees along a climatic moisture gradient

Tradeoffs between the energetic benefits and costs of traits can shape species and trait distributions along environmental gradients. Here we test predictions based on such tradeoffs using survival, growth, and 50 photosynthetic, hydraulic, and allocational traits of ten Eucalyptus species grown in four common gardens along an 8-fold gradient in precipitation/pan evaporation (P/Ep) in Victoria, Australia. Phylogenetically structured tests show that most trait-environment relationships accord qualitatively with theory. Most traits appear adaptive across species within gardens (indicating fixed genetic differences) and within species across gardens (indicating plasticity). However, species from moister climates have lower stomatal conductance than others grown under the same conditions. Responses in stomatal conductance and five related traits appear to reflect greater mesophyll photosynthetic sensitivity of mesic species to lower leaf water potential. Our data support adaptive cross-over, with realized height growth of most species exceeding that of others in climates they dominate. Our findings show that pervasive physiological, hydraulic, and allocational adaptations shape the distributions of dominant Eucalyptus species along a subcontinental climatic moisture gradient, driven by rapid divergence in species P/Ep and associated adaptations.

Copper bioconcentration and bioaccumulation in different eucalyptus species and their impact on plant physiology

Some plant species can bioaccumulate copper in their tissues and can be used for phytoremediation in contaminated soils. This work aimed to determine the bioconcentration and bioaccumulation of copper in eucalyptus species and how they affect the physiology of plants. The experimental design was completely randomized in a factorial arrangement (4 × 6), with four species of eucalyptus (Eucalyptus grandis W. Hill ex. Maiden, Corymbia citriodora (Hook.) K.D. Hill and L.A.S. Johnson, Eucalyptus saligna Smith and Eucalyptus dunnii Maiden), six doses of copper (zero, 80, 160, 240, 320, 400 mg kg−1), with twelve repetitions, carried out for 120 days in a greenhouse condition. The exotic species showed a high potential for growing in soils with high copper concentrations and also a potential for copper bioremediation. The highest copper concentration accumulation was shown mainly in the root system of the seedlings of Eucalyptus grandis, Eucalyptus saligna, Eucalyptus dunnii and Corymbia citriodora. E. saligna the copper toxicity was less pronounced, and the plants showed better physiologic effects. Cu levels increased with the doses, with greater intensity for species E. grandis and E. saligna with values of 21.1 and 15.5 mg kg−1, respectively. Also, the species showed a high capacity for copper bioaccumulation in the root system and showed high characteristics for phytoaccumulation and phytoextraction. So, these plants have great characteristics to grow in copper-contaminated sites being useful to reduce free copper in the environment.

Eco-extraction process of essential oil of the vegetal species Eucalyptus Camaldulensis Dehnh.: parametric modeling and optimization by Box-Behnken plan

This study was aimed to implement response surface methodology to optimize the hydrodistillation parameters of the essential oil of the plant species Eucalyptus camaldulensis (RSM). The distillation time, condensation flow rate, and particle size were the desired parameters. To generate factor combinations, a three-factor Box-Behnken design was used. The response surface analysis yielded a linear model, which was used to calculate the top yield of extracted oil based on the optimized conditions. ANOVA shows that the generated polynomial model was highly noteworthy, with R2=0.861. The peak yield was 0.513663 % at the optimal conditions, which were 1mm as particle size, 3.4 mL/min as condensation flow rate, and 210 min as extraction time. The GC/MS analysis revealed the presence of p-cymene (26.22 %), spathulenol (16.71 %), and 1.8-cineole (14.44 %) as major components.

Chemical Composition, Antibacterial Properties, and Anti-Enzymatic Effects of Eucalyptus Essential Oils Sourced from Tunisia.

This study was conducted to examine the chemical composition of the essential oils (EOs) from six Tunisian Eucalyptus species and to evaluate their anti-enzymatic and antibiofilm activities. The EOs were obtained through hydro-distillation of dried leaves and subsequently analyzed using GC/MS. The main class of compounds was constituted by oxygenated monoterpenes, particularly prominent in E. brevifolia (75.7%), E. lehmannii (72.8%), and E. woollsiana (67%). Anti-enzymatic activities against cholinesterases, α-amylase, and α-glucosidase were evaluated using spectrophotometric methods. Notably, the E. brevifolia, E. extensa, E. leptophylla, E. patellaris, and E. woollsiana EOs displayed potent acetylcholinesterase (AChE) inhibition (IC50: 0.25-0.60 mg/mL), with E. lehmannii exhibiting lower activity (IC50: 1.2 mg/mL). E. leptophylla and E. brevifolia showed remarkable α-amylase inhibition (IC50: 0.88 mg/mL), while E. brevifolia and E. leptophylla significantly hindered α-glucosidase (IC50 < 30 mg/mL), distinguishing them from other EOs with limited effects. Additionally, the EOs were assessed for their anti-biofilm properties of Gram-positive (Staphylococcus aureus and Listeria monocytogenes) and Gram-negative (Acinetobacter baumannii, Pseudomonas aeruginosa and Escherichia coli) bacterial strains. The E. extensa EO demonstrated the main antibiofilm effect against E. coli and L. monocytogenes with an inhibition > 80% at 10 mg/mL. These findings could represent a basis for possible further use of Eucalyptus EOs in the treatment of human microbial infections and/or as a coadjutant in preventing and treating Alzheimer's disease and/or diabetes mellitus.

Effects of Plant Growth Promoting Rhizospheric Bacteria (PGPR) on Survival, Growth and Rooting Architecture of Eucalyptus Hybrid Clones

Clonal plantation involves the rooting of cuttings from superior genotypes selected for their hybrid vigor and desired qualities. However, the cuttings of some Eucalyptus species and their hybrid genotypes present difficulties in their rooting capacity. Applying PGPR to cutting growth medium as a root stimulating agent has not been extensively studied for Eucalyptus tree species. We aimed to assess the rooting capacity of cuttings taken from two poor-rooting Eucalyptus hybrid clones of E. grandis × E. nitens through the application of PGPR in nursery trials. Seven rhizospheric bacterial species that demonstrated the ability to produce indole-3-acetic acid and to solubilise phosphate were used to prepare two rhizospheric consortium inoculums in which Pseudomonas-Bacillus strains and non-Pseudomonas-Bacillus were grouped. Inoculums were tested for their rooting stimulating capacity on cuttings of the hybrids GN 018B and GN 010 and compared to the nursery standard indole-3-butyric acid. A total of 320 cuttings were treated. Both hybrid clones demonstrated significant (p &lt; 0.0001) genotype differences for all three growth responses, i.e., total, root, and shoot length. Cuttings of both hybrids demonstrated high survival rates and rooting percentage. Although several rooting architectural configurations were prevalent, the Pseudomonas-Bacillus consortium promoted adventitious root development and fibrosity in GN 018B hybrids.

Termites infestation on different Eucalyptus wood species and control using natural oil from plants

Termites infestation on different Eucalyptus wood species and control using natural oil from plants

Leaf relative water content at 50% stomatal conductance measured by noninvasive NMR is linked to climate of origin in nine species of eucalypt.

Stomata are the gatekeepers of plant water use and must quickly respond to changes in plant water status to ensure plant survival under fluctuating environmental conditions. The mechanism for their closure is highly sensitive to disturbances in leaf water status, which makes isolating their response to declining water content difficult to characterise and to compare responses among species. Using a small-scale non-destructive nuclear magnetic resonance spectrometer as a leaf water content sensor, we measure the stomatal response to rapid induction of water deficit in the leaves of nine species of eucalypt from contrasting climates. We found a strong linear correlation between relative water content at 50% stomatal conductance (RWCgs50 ) and mean annual temperature at the climate of origin of each species. We also show evidence for stomata to maintain control over water loss well below turgor loss point in species adapted to warmer climates and secondary increases in stomatal conductance despite declining water content. We propose that RWCgs50 is a promising trait to guide future investigations comparing stomatal responses to water deficit. It may provide a useful phenotyping trait to delineate tolerance and adaption to hot temperatures and high leaf-to-air vapour pressure deficits.

Chemical Composition of Essential Oils from Eight Tunisian Eucalyptus Species and Their Antifungal and Herbicidal Activities.

Eucalyptus species are known to produce metabolites such as essential oils (EOs) that play an important role in the control of weeds, pests and phytopathogenic fungi. The aims of this study were as follows: (i) to determine the chemical composition of the EOs derived from eight Eucalyptus species growing in Tunisia, and (ii) to study their possible antifungal and herbicidal activities. EOs were obtained by hydrodistillation from the dried leaves of eight Eucalyptus species, namely, E. angulosa, E. cladocalyx, E. diversicolor, E. microcoryx, E. ovata, E. resinifera, E. saligna and E. sargentii, and the determination of their composition was achieved by GC and GC-MS. The EOs' antifungal activities were tested against four Fusarium strains, and the EOs' herbicidal properties were evaluated on the germination and seedling growth of three annual weeds (Trifolium campestre, Lolium rigidum and Sinapis arvensis) and three cultivated crop species (Lepidium sativum, Raphanus sativus and Triticum durum). The EO yields ranged between 0.12 and 1.32%. The most abundant components found were eucalyptol, α-pinene, p-cymene, trans-pinocarveol, α-terpineol and globulol. All EOs showed significant antifungal activity against the four phytopathogenic Fusarium strains. E. cladocalyx EO exhibited the highest level of antifungal activity, and the greatest inhibition of seed germination was obtained even at lowest concentrations used. These findings suggested that E. resinifera, E. ovata and E. cladocalyx EOs could have applications in agriculture as possible biopesticides, as Fusarium antagonists and as bioherbicides.

Using Transferable Eucalypt Microsatellite Markers to Identify QTL for Resistance to Ceratocystis Wilt Disease in Eucalyptus pellita F. Muel. (Myrtales, Myrtaceae)

The deployment of Eucalyptus pellita trees that are resistant to Ceratocystis manginecans is essential for the commercial plantations and therefore the sustainability of forest industries in Southeast Asia that utilize this resource. Current screening procedures are time-consuming and expensive but could be expedited with the aid of marker-assisted selection and breeding. The identification of genotypes with resistance to the disease may be facilitated if microsatellite markers developed in other Eucalyptus species are transferable and can be linked to quantitative trait loci (QTL) for disease resistance. This possibility was tested in 111 full-sib progenies and their parents by genotyping with 49 microsatellite markers developed in other Eucalyptus species. Disease development was assessed after stem inoculation with C. manginecans isolate Am60C. The disease index (DI) varied from 0 to 20% of stem length. There was a continuous distribution of resistant and susceptible seedlings with 60% in the resistant category. Of the 30 acceptable markers, 17 (56%) defined two linkage groups (LG). In each LG, one QTL with a significant logarithm of odds (LODs &gt; 13) was identified. The transferability of microsatellite markers developed in other Eucalyptus species facilitated the rapid identification of LGs and QTLs in E. pellita. To further refine the linkage map, the testing of more microsatellite markers and a larger population of progenies are required.

A Comprehensive Review Uncovering the Challenges and Advancements in the In Vitro Propagation of Eucalyptus Plantations.

The genus Eucalyptus is a globally captivated source of hardwood and is well known for its medicinal uses. The hybrid and wild species of Eucalyptus are widely used as exotic plantations due to their renowned potential of adapting to various systems and sites, and rapid large-scale propagation of genetically similar plantlets, which further leads to the extensive propagation of this species. Tissue culture plays a crucial role in the preservation, propagation, and genetic improvement of Eucalyptus species. Despite unquestionable progression in biotechnological and tissue culture approaches, the productivity of plantations is still limited, often due to the low efficiency of clonal propagation from cuttings. The obtained F1 hybrids yield high biomass and high-quality low-cost raw material for large-scale production; however, the development of hybrid, clonal multiplication, proliferation, and post-developmental studies are still major concerns. This riveting review describes the problems concerning the in vitro and clonal propagation of Eucalyptus plantation and recent advances in biotechnological and tissue culture practices for massive and rapid micropropagation of Eucalyptus, and it highlights the Eucalyptus germplasm preservation techniques.

Characterisation of Chrysoporthe cubensis and Chrysoporthe deuterocubensis, the Stem Canker Diseases of Eucalyptus spp. in a Forest Plantation in Malaysia

Commercial plantations of Eucalyptus species have been established in Malaysia, especially during the past 10 years, with the aim of sustaining the supply of wood and timber products for industrial use in Malaysia. As part of an assessment of fungal diseases affecting Eucalyptus species in four regions in Malaysia, including Kelantan, Pahang, Sabah, and Selangor, stem canker disease was discovered to be a widespread disease infecting Eucalyptus species in Malaysia. This study aimed to identify the fungus-causing stem canker disease, test its pathogenicity in Eucalyptus, and determine the mating type of isolates from the infected trees. The fungi were identified based on morphology and through comparisons of DNA sequence data from the ITS, β-tubulin 2 gene, and TEF-1α gene regions. Phylogenetic analyses showed that the causal agent of the stem canker was Chrysoporthe cubensis infecting Eucalyptus plantations in Pahang and Chrysoporthe deuterocubensis infecting Eucalyptus plantations in Kelantan, Sabah, and Selangor. We believe this is the first report of Chrysoporthe cubensis-infected Eucalyptus in Malaysia and Southeast Asia, while Chrysoporthe deuterocubensis is the first-reported species infecting Eucalyptus pellita in Malaysia. Moreover, the fact that the mating-type MAT1-1 and MAT1-2 genes and the pheromone genes ppg1, ppg2, pre1, and pre2 were identified in all isolates indicates that Chrysoporthe cubensis and Chrysoporthe deuterocubensis are homothallic mating systems. Pathogenicity was tested on a 3-year-old standing tree, 1-year-old seedling, and detached healthy leaves, which were re-isolated for fulfilling Koch’s postulates. In pathogenicity trials, both Chrysoporthe cubensis and Chrysoporthe deuterocubensis gave rise to lesions on wounded Eucalyptus. Both Chrysoporthe spp. were equally pathogenic to Eucalyptus urograndis and Eucalyptus pellita and should be regarded as a biosecurity concern in Malaysia’s forest plantation industry.

Genome-wide analysis highlights genetic admixture in exotic germplasm resources of Eucalyptus and unexpected ancestral genomic composition of interspecific hybrids.

Eucalyptus is an economically important genus comprising more than 890 species in different subgenera and sections. Approximately twenty species of subgenus Symphyomyrtus account for 95% of the world's planted eucalypts. Discrimination of closely related eucalypt taxa is challenging, consistent with their recent phylogenetic divergence and occasional hybridization in nature. Admixture, misclassification or mislabeling of Eucalyptus germplasm resources maintained as exotics have been suggested, although no reports are available. Moreover, hybrids with increased productivity and traits complementarity are planted worldwide, but little is known about their actual genomic ancestry. In this study we examined a set of 440 trees of 16 different Eucalyptus species and 44 interspecific hybrids of multi-species origin conserved in germplasm banks in Brazil. We used genome-wide SNP data to evaluate the agreement between the alleged phylogenetic classification of species and provenances as registered in their historical records, and their observed genetic clustering derived from SNP data. Genetic structure analyses correctly assigned each of the 16 species to a different cluster although the PCA positioning of E. longirostrata was inconsistent with its current taxonomy. Admixture was present for closely related species' materials derived from local germplasm banks, indicating unintended hybridization following germplasm introduction. Provenances could be discriminated for some species, indicating that SNP-based discrimination was directly proportional to geographical distance, consistent with an isolation-by-distance model. SNP-based genomic ancestry analysis showed that the majority of the hybrids displayed realized genomic composition deviating from the expected ones based on their pedigree records, consistent with admixture in their parents and pervasive genome-wide directional selection toward the fast-growing E. grandis genome. SNP data in support of tree breeding provide precise germplasm identity verification, and allow breeders to objectively recognize the actual ancestral origin of superior hybrids to more realistically guide the program toward the development of the desired genetic combinations.

Variation in Growth and Wood Properties of Four Eucalyptus Species, their Hybrids and Clones in Vidarbha Region, Maharashtra, India

Variation in Growth and Wood Properties of Four Eucalyptus Species, their Hybrids and Clones in Vidarbha Region, Maharashtra, India

Correlating Eucalyptus leaf metabolomics with preference of the bronze bug, Thaumastocoris peregrinus.

Eucalyptus species are among the most planted trees in forestry production, an ever-increasing commercial activity worldwide. Forestry expansion demands a continuous search for preventive and sanitary measures against pests and diseases. Massive application of phytosanitary products is incompatible with the forestry sector, so forest health management must be based on other principles. In this context, studies on insect plant relationships mediated by plant metabolites may contribute information relevant to plant resistance and genotype selection. In this study, we analyzed the leaf metabolome of four Eucalyptus species commonly planted in southern South America, to correlate this chemical information with feeding preference of Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae), an important pest of eucalypt plantations. Gas chromatography mass spectrometry analyses were performed on polar and non-polar leaf extracts from Eucalyptus globulus, Eucalyptus grandis, Eucalyptus robusta, and Eucalyptus tereticornis (Myrtaceae). Feeding preferences were assessed in two-choice laboratory bioassays resulting in a preference gradient of the four plant species. Moreover, a performance bioassay where we contrasted survival and development time between the most and least preferred plants, showed a clear correlation with preference both in survival and developmental time of the most susceptible nymph instar. We found that species with high or low feeding preferences differ significantly in several foliar metabolites, which may be acting as feeding stimulants or deterrents for T. peregrinus. These findings may provide useful criteria for choosing Eucalyptus genotypes when planting in bronze bug infested areas.

Whole-Genome-Sequence-Based Classification of Xanthomonas euvesicatoria pv. eucalypti and Computational Analysis of the Type III Secretion System.

Xanthomonas spp. infect a wide range of annual and perennial plants. Bacterial blight in young seedlings of Eucalyptus spp. in Indonesia was originally identified as X. perforans. However, these strains failed to elicit a hypersensitive response (HR) on either tomatoes or peppers. Two of the strains, EPK43 and BCC 972, when infiltrated into tomato and pepper leaves, failed to grow to significant levels in comparison with well-characterized X. euvesicatoria pv. perforans (Xp) strains. Furthermore, spray inoculation of 'Bonny Best' tomato plants with a bacterial suspension of the Eucalyptus strains resulted in no obvious symptoms. We sequenced the whole genomes of eight strains isolated from two Eucalyptus species between 2007 and 2015. The strains had average nucleotide identities (ANIs) of at least 97.8 with Xp and X. euvesicatoria pv. euvesicatoria (Xeu) strains, both of which are causal agents of bacterial spot of tomatoes and peppers. A comparison of the Eucalyptus strains revealed that the ANI values were >99.99% with each other. Core genome phylogeny clustered all Eucalyptus strains with X. euvesicatoria pv. rosa. They formed separate clades, which included X. euvesicatoria pv. alangii, X. euvesicatoria pv. citrumelonis, and X. euvesicatoria pv. alfalfae. Based on ANI, phylogenetic relationships, and pathogenicity, we designated these Eucalyptus strains as X. euvesicatoria pv. eucalypti (Xee). Comparative analysis of sequenced strains provided unique profiles of type III secretion effectors. Core effector XopD, present in all pathogenic Xp and Xeu strains, was absent in the Xee strains. Comparison of the hrp clusters of Xee, Xp, and Xeu genomes revealed that HrpE in Xee strains was very different from that in Xp and Xeu. To determine if it was functional, we deleted the gene and complemented with the Xee hrpE, confirming it was essential for secretion of type III effectors. HrpE has a hypervariable N-terminus in Xanthomonas spp., in which the N-terminus of Xee strains differs significantly from those of Xeu and Xp strains.