Rust is one of the most-damaging eucalypt diseases in Brazil and is considered a potential threat to eucalypt plantations worldwide. To determine the mode of inheritance of resistance in the Eucalyptus grandis- Puccinia psidii pathosystem, ten full-sib families, generated from crosses between susceptible and resistant trees, were inoculated with a single-pustule isolate of the pathogen and rust severity was scored. The observed segregation ratios in segregating families suggested major gene control of rust resistance, although clearly incomplete penetrance, variable expressivity and minor genes are also involved in the global rust-resistance response. To identify markers linked to the resistance locus, screening of RAPD polymorphisms was conducted using bulked segregant analysis in a large full-sib family. A linkage group was built around the Ppr1 gene ( P. psidii resistance gene 1) encompassing six RAPD markers, with a genetic window spanning 5 cM with the two most-closely linked flanking markers. Besides these two flanking markers, RAPD marker AT9/917 co-segregated with Ppr1 without a single recombinant in 994 meioses. This tightly linked marker should prove useful for marker-assisted introgression and will provide an initial lead for a positional cloning effort of this resistance allele. This is the first report of a disease resistance gene identified in Eucalyptus, and one of the few examples of the involvement of a major gene in a non-coevolved pathosystem.

The diversity and beneficial characteristics of endophytic microorganisms have been studied in several host plants. However, information regarding naturally occurring seed-associated endophytes and vertical transmission among different life-history stages of hosts is limited. Endophytic bacteria were isolated from seeds and seedlings of 10 Eucalyptus species and two hybrids. The results showed that endophytic bacteria, such as Bacillus, Enterococcus, Paenibacillus and Methylobacterium, are vertically transferred from seeds to seedlings. In addition, the endophytic bacterium Pantoea agglomerans was tagged with the gfp gene, inoculated into seeds and further reisolated from seedlings. These results suggested a novel approach to change the profile of the plants, where the bacterium is a delivery vehicle for desired traits. This is the first report of an endophytic bacterial community residing in Eucalyptus seeds and the transmission of these bacteria from seeds to seedlings. The bacterial species reported in this work have been described as providing benefits to host plants. Therefore, we suggest that endophytic bacteria can be transmitted vertically from seeds to seedlings, assuring the support of the bacterial community in the host plant.

Recent advances in genomics and proteomics have provided an excellent opportunity to understand complex biological processes such as wood formation at the gene and protein levels. The aim of this work was to describe the proteins participating in the processes involved in juvenile wood formation by isolating proteins from the cambial region of Eucalyptus grandis, at three ages of growth (6-month-old seedlings, 3- and 6-year-old trees), and also to identify proteins differentially expressed. Using a 2-D-LC-MS/MS strategy we identified a total of 240 proteins, with 54 corresponding spots being present in at least two ages. Overall, nine proteins classified into the functional categories of metabolism, cellular processes, and macromolecular metabolism showed significant changes in expression. Proteins were classified into seven main functional categories, with metabolism representing 35.2% of the total proteins identified. The comparison of the reference maps showed not only differences in the expression pattern of individual proteins at each age, but also among isoforms. The results described in this paper provide a dynamic view of the proteins involved in the formation of juvenile wood in E. grandis.

Despite the importance of Eucalyptus spp. in the pulp and paper industry, functional genomic approaches have only recently been applied to understand wood formation in this genus. We attempted to establish a global view of gene expression in the juvenile cambial region of Eucalyptus grandis Hill ex Maiden. The expression profile was obtained from serial analysis of gene expression (SAGE) library data produced from 3- and 6-year-old trees. Fourteen-base expressed sequence tags (ESTs) were searched against public Eucalyptus ESTs and annotated with GenBank. Altogether 43,304 tags were generated producing 3066 unigenes with three or more copies each, 445 with a putative identity, 215 with unknown function and 2406 without an EST match. The expression profile of the juvenile cambial region revealed the presence of highly frequent transcripts related to general metabolism and energy metabolism, cellular processes, transport, structural components and information pathways. We made a quantitative analysis of a large number of genes involved in the biosynthesis of cellulose, pectin, hemicellulose and lignin. Our findings provide insight into the expression of functionally related genes involved in juvenile wood formation in young fast-growing E. grandis trees.

International audienceAbstract Key messageThe identification of an anomaly with deleterious effect and Mendelian segregation (3 normal: 1 abnormal) allowed the detection of a marker linked to a locus for the anomalous phenotype. This marker could be useful in marker-assisted breeding programs and for understanding this anomaly. ContextDue to the mixed mating systems in Eucalyptus genera, outcrossing populations exhibit low inbreeding depression (abnormalities). In this study, we identified a developmental anomaly in a controlled cross of Eucalyptus grandis, with Mendelian segregation. The anomaly was characterized by branching, reduced height, smaller individual leaf area, and asymmetrical leaf shape. Aims and methodsTo investigate and evaluate the anomaly detected, we performed morphological and molecular analysis. The genitors and bulks of normal and abnormal individuals were screened with random primers. The polymorphic markers that co-segregated with the anomalous phenotype were selected, validated, converted into SCAR markers and analyzed in silico. ResultsMorphological analysis showed significant differences between normal and abnormal phenotypes. The co-segregating marker was present in one genitor, in all abnormal individuals, and in 31 % of normal individuals (recombinants). In silico analyses revealed a mutation of two base pairs between the contrasting phenotypes and identified the marker in an intergenic region presenting partial identity with an expressed sequence tag (EST) of Bet v1-like genes. ConclusionA marker was developed to identify the recessive allele for the anomaly in E. grandis. This will be important for management of crosses in eucalypt breeding programs and in anomaly studies

Eucalyptus grandis Hill ex Maiden and its hybrids are commonly planted by the Brazilian pulp and paper industry, but they are the most susceptible to the neotropical rust disease caused by Puccinia psidii Winter. In an initial attempt to understand the mechanisms of resistance, we constructed two contrasting Serial Analysis of Gene Expression (SAGE) libraries using susceptible and resistant individuals from a segregating half-sibling E. grandis population. Using the Z-test we identified tags differentially expressed between the libraries, preferentially 239 in the susceptible and 232 in the resistant type individuals. Using public (Expressed Sequence Tags) EST databases, 40 of the susceptible and 70 of the resistant tags matched ESTs and were annotated. By comparing the type of genes and their expression levels, distinct differences between the libraries were observed. Susceptible plants showed gene expression linked to leaf senescence, generalised stress responses and detoxification, and are apparently incapable of inducing a competent host defence response. On the other hand, resistant plants showed genes upregulated for cellular polarisation, cytoskeleton restructuring, vesicle transport, and cellulose and lignin biosynthesis. In the resistant individuals, evidence for systemic resistance, anti-oxidative responses and a hypersensitive response was also observed, although no R gene was identified.

O presente estudo empregou o procedimento de envelhecimento artificial, submetendo sementes por 96 horas à 42° C e 100% de UR para compará-las com sementes estocadas por 5, 10 e 15 anos e utilizou eletroforese de isoenzimas e atividade de peroxidase por método colorimétrico. Houve um decréscimo significante na germinação das sementes envelhecidas artificialmente, quando comparadas com as sementes estocadas. A atividade da peroxidase foi baixa, porém houve uma curva ascendente que acompanhou a idade das sementes. Observou-se um aumento nas atividades da malato desidrogenase e a-esterase 2 e 3 e uma diminuição acentuada para a-esterase 1 e baixa para a fosfatase ácida. Os resultados acompanharam o aumento do tempo de estocagem das sementes e do estresse que ocorre por causa das condições de temperatura e umidade empregadas no processo de envelhecimento artificial.

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