Transcriptome analysis of Eucalyptus grandis genotypes reveals constitutive overexpression of genes related to rust (Austropuccinia psidii) resistance.

Key Message A resistant E. grandis genotype showed a constitutive overexpression of genes related to resistance to myrtle rust caused by A. psidii. Abstract Myrtle rust caused by Austropuccinia psidii is considered one of the most important fungal diseases affecting Eucalyptus spp. plantations in Brazil. Although the selection and planting of resistant eucalypt genotypes have been the major strategies to manage the disease in Brazil, the molecular mechanisms involved in resistance are still unclear. In this study, we evaluated the gene expression profile of two contrasting Eucalyptus grandis genotypes in resistance level to rust by RNA-Seq. The two genotypes showed a very different background gene expression level even without A. psidii infection. The resistant genotype had a constitutive overexpression of a large number of protein-coding genes compared to the susceptible genotype. These genes were mainly associated with signal transduction, photosynthesis, regulation and response to salicylic acid (SA), and protein kinase leucine-rich receptors (PK-LRR). PK-LRR and SA mediated disease resistance are well known to be effective against obligate biotroph pathogens, such as A. psidii. In addition, at 24 h after infection, the susceptible genotype was able to activate some response, however, several resistance-related proteins had their expression level reduced with A. psidii infection. Here, we present the first analysis of E. grandis genotypes transcriptomes infected by A. psidii and it reveals a constitutive overexpression of several resistance-related genes in the resistant genotype compared to the susceptible one. Our findings have the potential to be used as candidate molecular markers for resistance to myrtle rust.

Genetically engineered eucalyptus expressing pesticidal proteins from Bacillus thuringiensis for insect resistance: a risk assessment evaluation perspective.

Eucalyptus covers approximately 7.5 million hectares in Brazil and serves as the primary woody species cultivated for commercial purposes. However, native insects and invasive pests pose a significant threat to eucalyptus trees, resulting in substantial economic losses and reduced forest productivity. One of the primary lepidopteran pests affecting eucalyptus is Thyrinteina arnobia (Stoll, 1782) (Lepidoptera: Geometridae), commonly referred to as the brown looper caterpillar. To address this issue, FuturaGene, the biotech division of Suzano S.A., has developed an insect-resistant (IR) eucalyptus variety, which expresses Cry pesticidal proteins (Cry1Ab, Cry1Bb, and Cry2Aa), derived from Bacillus thuringiensis (Bt). Following extensive safety assessments, including field trials across various biomes in Brazil, the Brazilian National Technical Commission of Biosafety (CTNBio) recently approved the commercialization of IR eucalyptus. The biosafety assessments involved the analysis of molecular genomics, digestibility, thermostability, non-target organism exposure, degradability in the field, and effects on soil microbial communities and arthropod communities. In addition, in silico studies were conducted to evaluate allergenicity and toxicity. Results from both laboratory and field studies indicated that Bt eucalyptus is as safe as the conventional eucalyptus clone for humans, animals, and the environment, ensuring the secure use of this insect-resistant trait in wood production.

Safety Assessment of the CP4 EPSPS and NPTII Proteins in Eucalyptus.

Glyphosate herbicide treatment is essential to sustainable Eucalyptus plantation management in Brazil. Eucalyptus is highly sensitive to glyphosate, and Suzano/FuturaGene has genetically modified eucalyptus to tolerate glyphosate, with the aim of both protecting eucalyptus trees from glyphosate application damage and improving weed management. This study presents the biosafety results of the glyphosate-tolerant eucalyptus event 751K032, which expresses the selection marker neomycin phosphotransferase II (NPTII) enzyme and CP4-EPSPS, a glyphosate-tolerant variant of plant 5-enolpyruvyl-shikimate-3-phosphate synthase enzyme. The transgenic genetically modified (GM) event 751K032 behaved in the plantations like conventional non-transgenic eucalyptus clone, FGN-K, and had no effects on arthropods and soil microorganisms. The engineered NPTII and CP4 EPSPS proteins were heat-labile, readily digestible, and according to the bioinformatics analyses, unlikely to cause an allergenic or toxic reaction in humans or animals. This assessment of the biosafety of the glyphosate-tolerant eucalyptus event 751K032 concludes that it is safe to be used for wood production.

Two new species of Calonectria (Hypocreales, Nectriaceae) causing Eucalyptus leaf blight in Brazil.

In recent decades, commercial Eucalyptus plantations have expanded toward the warm and humid regions of northern and northeastern Brazil, where Calonectria leaf blight (CLB) has become the primary fungal leaf disease of this crop. CLB can be caused by different Calonectria species, and previous studies have indicated that Calonectria might have high species diversity in Brazil. During a disease survey conducted in three commercial plantations of Eucalyptus in northeastern Brazil, diseased leaves from Eucalyptus trees with typical symptoms of CLB were collected, and Calonectria fungi were isolated. Based on phylogenetic analyses of six gene regions (act, cmdA, his3, rpb2, tef1, and tub2) and morphological characteristics, two new species of Calonectria were identified. Five isolates were named as C.paragominensis sp. nov. and four were named as C.imperata sp. nov. The pathogenicity to Eucalyptus of both species was confirmed by fulfilling the Koch's postulates.

First Draft Genome Sequence of Xanthomonas axonopodis pv. eucalyptorum, Causal Agent of Bacterial Leaf Blight on Eucalypt.

Here, we report the annotated draft genome sequence of Xanthomonas axonopodis pv. eucalyptorum pathotype strain LPF602 (synonym Xanthomonas axonopodis BSC45a), isolated from eucalypt leaves showing bacterial blight symptoms in Brazil. The availability of these genomic data will help improve the understanding of the evolution and molecular mechanisms involved in the pathogenesis of this microorganism.

Draft Genome Sequence of Erwinia psidii, Causal Agent of Bacterial Blight of Guava (Psidium guava) and Dieback of Eucalypt (Eucalyptus spp.).

Here, we present a draft genome sequence of the type strain IBSBF 435 of Erwinia psidii (Enterobacteriaceae), a phytopathogen that causes bacterial blight on guava (Psidium guava) and dieback and wilt on eucalypt (Eucalyptus spp.), both of which are important emerging diseases.

Eucalypt plants are physiologically and metabolically affected by infection with Ceratocystis fimbriata.

Ceratocystis wilt, caused by Ceratocystis fimbriata, is currently one of the most important disease in eucalypt plantations. Plants infected by C. fimbriata have lower volumetric growth, lower pulp yields and reduced timber values. The physiological bases of infection induced by this pathogen in eucalypt plant are not known. Therefore, this study aims to assess the physiological and metabolic changes in eucalypt clones that are resistant and susceptible to C. fimbriata. Once, we evaluated in detail their leaf gas exchange, chlorophyll a fluorescence, water potential, metabolite profiling and growth-related parameters. When inoculated, the susceptible clone displayed reduced water potential, CO2 assimilation rate, stomatal conductance, transpiration rate, photochemical quenching coefficient, electron transport rate, and root biomass. Inoculated resistant and susceptible clones both presented higher respiration rates than healthy plants. Many compounds of primary and secondary metabolism were significantly altered after fungal infection in both clones. These results suggest that, C. fimbriata interferes in the primary and secondary metabolism of plants that may be linked to the induction of defense mechanisms and that, due to water restrictions caused by the fungus in susceptible plants, there is a partial closure of the stomata to prevent water loss and a consequent reduction in photosynthesis and the transpiration rate, which in turn, leads to a decrease in the plant's growth-related. These results combined, allowed for a better understanding of the physiological and metabolic changes following the infectious process of C. fimbriata, which limit eucalypt plant growth.

Rhizobacterial characterization for quality control of eucalyptus biogrowth promoter products.

Plant growth-promoting rhizobacteria strains from special formulations have been used to optimize eucalyptus cutting production. To undertake quality control for the formulated products, the rhizobacterial strains should be characterized to assess their purity and authentication. In the present study, we characterized nine strains of rhizobacteria, including three Bacillus subtilis (S1, S2 and 3918), two Pseudomonas sp. (MF4 and FL2), P. putida (MF2), P. fulva (Ca), Frateuria aurantia (R1), and Stenotrophomonas maltophilia (CIIb). The strains were differentiated by colony morphology after 24h of incubation in three different solid state culture media (glucose-nutritive agar, 523 medium and yeast extract-mannitol agar), sensitivity to a panel of 28 antibiotics (expressed according to the formation of inhibition halos of bacterial growth in the presence of antibiotics), and PCR-RFLP profiles of the 16S rDNA gene produced using nine restriction enzymes. It was possible to differentiate all nine strains of rhizobacteria using their morphological characteristics and sensitivity to antibiotics. The molecular analysis allowed us to separate the strains CIIb, FL2 and R1 from the strains belonging to the genera Bacillus and Pseudomonas. By using these three methods concomitantly, we were able to determine strain purity and perform the authentication.

Rhizobacterial characterization for quality control of eucalyptus biogrowth promoter products

Abstract Plant growth-promoting rhizobacteria strains from special formulations have been used to optimize eucalyptus cutting production. To undertake quality control for the formulated products, the rhizobacterial strains should be characterized to assess their purity and authentication. In the present study, we characterized nine strains of rhizobacteria, including three Bacillus subtilis (S1, S2 and 3918), two Pseudomonas sp. (MF4 and FL2), P. putida (MF2), P. fulva (Ca), Frateuria aurantia (R1), and Stenotrophomonas maltophilia (CIIb). The strains were differentiated by colony morphology after 24 h of incubation in three different solid state culture media (glucose-nutritive agar, 523 medium and yeast extract-mannitol agar), sensitivity to a panel of 28 antibiotics (expressed according to the formation of inhibition halos of bacterial growth in the presence of antibiotics), and PCR-RFLP profiles of the 16S rDNA gene produced using nine restriction enzymes. It was possible to differentiate all nine strains of rhizobacteria using their morphological characteristics and sensitivity to antibiotics. The molecular analysis allowed us to separate the strains CIIb, FL2 and R1 from the strains belonging to the genera Bacillus and Pseudomonas. By using these three methods concomitantly, we were able to determine strain purity and perform the authentication.

Caracterização dos danos causados por Heilipodus naevulus em plantios de eucalipto no Espírito Santo, Brasil / Characterization of damages caused by Heilipodus naevulus in eucalypt plantations on Espírito Santo, Brazil

O objetivo do presente trabalho foi avaliar os efeitos dos danos causados por Heilipodus naevulus Mann. (Coleoptera: Curculionidae) no desenvolvimento de plantios jovens de eucalipto, além de relatar, pela primeira vez, a ocorrência desse inseto no estado do Espírito Santo. O período crítico para ocorrência foi caracterizado entre outubro a dezembro, de acordo com o regime de chuvas. Os danos causados pela praga reduziram o desenvolvimento e a qualidade das árvores de eucalipto.

This research aimed to evaluate the effects of damages caused by Heilipodus naevulus Mann. (Coleoptera: Curculionidae) to the development of eucalyptus young plantation, and to record, for the first time, the occurrence on Espírito Santo State, Brazil. The critical period of occurrence was observed between October to December according with rain regime. The damages caused by the pest reduced the development and quality of eucalyptus trees.

Primeiro registro de Chalcodermus bicolor (Coleoptera: Curculionidae) em plantios de eucalipto / First record of Chalcodermus bicolor in eucalypt plantations

Este trabalho objetivou relatar, pela primeira vez, o ataque de um besouro podador em plantios de clones híbridos de eucalipto (Eucalyptus urophylla vs. E. grandis), localizados nos Estados da Bahia e do Espírito Santo. A espécie foi determinada como sendo Chalcodermus bicolor Fiedler, 1936 (Curculionidae: Molytinae). A fêmea poda os ponteiros e constrói um pequeno orifício onde deposita um único ovo, logo abaixo do ponto de incisão. O eucalipto é o primeiro hospedeiro relatado para esta espécie de besouro.

This research aimed to record, for the first time, the damage caused by a pruner beetle on hybrid eucalypts cloned trees (Eucalyptus urophylla vs. E. grandis), located in Bahia and Espírito Santo States, Brazil. The specie was determined as Chalcodermus bicolor Fiedler, 1936 (Curculionidae: Molytinae). The female prunes the tree shoot and lays a single egg inside a small hole, just below the incision point. Eucalypt is the first host recorded to this pruner beetle specie.

Rhizobacterial promotion of eucalypt rooting and growth

A total of 107 rhizobacterial isolates, obtained from the rhizosphere of eucalypt clones were tested as rooting inducers of cuttings and mini-cuttings planted in substrate composed of carbonized rice husk and vermiculite (1:1). Cuttings and mini-cuttings were planted in conical plastic tubes containing treated and untreated (control) substrate and kept under intermittent mist irrigation at 26-28°C. After 35 days, rooting percentage and dry root matter of cuttings were evaluated. Ten isolates capable of providing gains of up to 110 percent in root formation and up to 250 percent in root biomass over non-inoculated control cuttings were selected. Gains in rooting varied according to clone and isolate tested. The greatest gains were obtained for the mini-cuttings exhibiting the lowest rooting efficiency. Among the ten isolates tested, only 3918 (code R98) and MF4 (code R87), produced 3-indole-acetic acid in vitro, at concentrations of 0.7 and 0.67 µg ml-1, respectively. Significant increases in rooting and root dry matter of cuttings grown on rhizobacteria-inoculated substrate were found when compared to untreated or indole-butyric acid (IBA) treated mini-cuttings.

Neste trabalho, testaram-se 107 rizobactérias, isoladas da rizosfera de mudas de clones de eucalipto, quanto ao seu potencial como promotoras de enraizamento de estacas e miniestacas de eucalipto, em substrato à base de casca de arroz carbonizada e vermiculita (1:1). Estacas e miniestacas foram plantadas em tubetes cônicos contendo substrato tratado e não tratado (testemunha) e foram mantidas sob nebulização intermitente de água a 26-28°C. Aos 35 dias, avaliou-se a porcentagem média de estacas enraizadas e a massa seca do sistema radicular. Dez isolados destacaram-se como indutores de enraizamento e crescimento, propiciando ganhos de até 110 por cento e de 250 por cento, respectivamente. Esses isolados também foram eficientes no enraizamento de miniestacas, cujos ganhos variaram de acordo com o clone e isolado testado. Os maiores incrementos obtidos no enraizamento de estacas foram superiores aos observados para miniestacas. Em geral, quanto menor o índice de enraizamento do clone, maior foi o ganho médio obtido com a inoculação. Apenas os isolados 3918 (código R98) e MF4 (código R87) foram capazes de produzir ácido indol-acético (AIA) in vitro, em quantidades equivalentes a 0,7 e 0,67 µg/ml de suspensão, respectivamente. Quando comparados ao tratamento de miniestacas em ácido indol butírico (AIB), estes isolados promoveram incrementos significativos na porcentagem de enraizamento e na massa seca do sistema radicular de miniestacas.