Lúcio Mauro da Silva Guimarães

Leaf gas exchange and chlorophyll a fluorescence of Eucalyptus urophylla in response to Puccinia psidii infection

One of the most important diseases of eucalyptus plantations is caused by the rust fungus Puccinia psidii. While the genetic basis of rust resistance has been addressed recently, little is known about the physiological aspects of Eucalyptus–P. psidii interaction. In order to fill this gap, we undertook a study investigating the effects of P. psidii infection on photosynthetic processes of two E. urophylla clones with contrasting resistance to the pathogen. Our results show that gas exchange and chlorophyll a fluorescence parameters were virtually unaffected in the resistant clone. In the susceptible clone, photosynthetic rates were chiefly constrained by biochemical limitations to carbon fixation. Photosynthesis was impaired only in symptomatic tissues since the reductions in photosynthetic rates were proportional to the diseased leaf area. Rust infection provoked chronic photoinhibition to photosynthesis in the susceptible clone. Overall, differences in the ability for light capture, use and dissipation may play a significant role in explaining the clonal differences in Eucalyptus in response to P. psidii infection. To our knowledge, this is the first report of the effect of rust infection on gas exchange and chlorophyll a fluorescence parameters in Eucalyptus.

Positioning of the major locus for Puccinia psidii rust resistance (Ppr1) on the Eucalyptus reference map and its validation across unrelated pedigrees

In this report the major locus for Puccinia psidii rust resistance, Ppr1, was positioned on the reference genetic map for Eucalyptus. Additionally, its position was validated by association genetics in a related and two unrelated pedigrees involving different Eucalyptus grandis resistant trees crossed to individuals of two other species, Eucalyptus tereticornis and Eucalyptus camaldulensis. These results are consistent with the hypothesis that Ppr1 controls a large proportion of the variation for rust resistance, strengthening its role as a major locus in Eucalyptus and providing its unequivocal genomic position on linkage group 3. A localized map with 19 microsatellite loci was built around Ppr1. Multiallelic profiles were observed at several mapped microsatellites suggesting recent tandem duplications in the genomic landscape surrounding Ppr1. Markers EMBRA125 and EMBRA1071 flank Ppr1 at 9.5% and 7% recombination, respectively, and were found to be in linkage equilibrium in a E. grandis breeding population, consistent with the expectations in outcrossed Eucalyptus. Their potential use for MAS will specifically be directed to identifying resistant offspring of P. psidii resistant parent trees that are heterozygous at Ppr1. In these circumstances, a significant amount of LD is expected to occur between specific alleles at flanking microsatellites and the resistance allele at Ppr1. Moreover, the positional information of Ppr1 paves the way for prospective undertakings in this genomic region with the upcoming availability of a draft genome for E. grandis.

Genetic mapping provides evidence for the role of additive and non-additive QTLs in the response of inter-specific hybrids of Eucalyptus to Puccinia psidii rust infection

Eucalypts are susceptible to a wide range of diseases. One of the most important diseases that affect Eucalyptus plantations worldwide is caused by the rust fungus Puccinia psidii. Here, we provide evidence on the complex genetic control of rust resistance in Eucalyptus inter-specific hybrids, by analyzing a number of full-sib families that display different patterns of segregation for rust resistance. These families are totally unrelated to those previously used in other inheritance studies of rust resistance. By using a full genome scan with 114 genetic markers (microsatellites and expressed sequence tag derived microsatellites) we also corroborated the existence and segregation of a resistance locus, explaining 11.5% of the phenotypic variation, on linkage group 3, corresponding to Ppr1. This find represents an additional validation of this locus in totally unrelated pedigree. We have also detected significant additive × additive digenic interactions with LOD >10.0 on several linkage groups. The additive and epistatic QTLs identified explain between 29.8 and 44.8% of the phenotypic variability for rust resistance. The recognition that both additive and non-additive genetic variation (epistasis) are important contributors to rust resistance in eucalypts reveals the complexity of this host-pathogen interaction and helps explain the success that breeding has achieved by selecting rust-resistant clones, where all the additive and non-additive effects are readily captured. The positioning of epistatic QTLs also provides starting points to look for the underlying genes or genomic regions controlling this phenotype on the upcoming E. grandis genome sequence.

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 (U) trees, as well as in 495 individuals from 27 progenies derived from crosses between the trees. In the field, six-months-old test seedlings were inoculated with C. cubensis. Lesion length in the xylem and bark was measured eight months later. The results demonstrated that xylem lesions could preferentially be used for the selection of resistant clones. Eight trees (7 U and 1 G) were susceptible, and the remainder (8 U and 4 G) resistant. Individual narrow and broad sense heritability estimates were 17 and 81%, respectively, thereby suggesting that canker resistance is quantitative and highly dependent on dominance and epistasis.

A new race of Puccinia psidii defeats rust resistance in eucalypt

Rust caused by Puccinia psidii is one the most destructive diseases of Eucalyptus. Management of the disease is achieved through selection of resistant host genotypes. Recently, eucalypt plants from clone BA6021, resistant to P. psidii isolate race-1, were infected by rust in Brazil. Microsatellite profiles of infected plants confirmed that the host was indeed clone BA6021. In pathogenicity tests, the resistant clones BA6021 and G21 (which carry the resistance gene Ppr-1) were found susceptible to the newly discovered isolate EUBA-1, indicating a new biotype of the pathogen. These results show that the isolate EUBA-1 and other potentially unrecognized pathogen races should be given strong consideration for eucalypt breeding programs aimed rust resistance.

Detection of QTL associated with rust resistance using IBD-based methodologies in exogamic Eucalyptus spp. populations

In Brazil the rust caused by Puccinia psidii Winter stands out as the most important disease of eucalyptus. The use of resistant genotypes is the main control method, which makes the detection of markers linked to rust resistance essential to the selection of resistant genotypes. In this study, an F1 progeny of 131 plants from interspecific crossings of Eucalyptus was used to identify markers linked to resistance genes for this pathogen. An integrated map was constructed for linkage group three based on microsatellite markers. For QTL mapping two methodologies based on alleles identical-by-descent (IBD) were used: single marker analysis of Haseman and Elston and the interval mapping procedure of Fulker and Cardon. Both methods showed significant association for the Embra 125 marker.The QTL that explained 42 % of the phenotypic variation was mapped to 0.02 cM of this marker by the Fulker and Cardon. Marker Embra 125 has potential use in assisted selection, thus increasing the efficiency of the selection of resistant genotypes.

Genetic variation, morphology and pathogenicity of Ceratocystis fimbriata on Hevea brasiliensis in Brazil

Ceratocystis fimbriata causes diseases on a wide variety of plants in Brazil, including rubber tree (Hevea brasiliensis), on which it causes gray mold or moldy rot on tapping panels affecting latex yield. However, C. fimbriata isolated from rubber tree have not been critically studied. Phylogenetic analyses of sequences of ITS rDNA and a mating type gene placed rubber tree isolates from Acre and Bahia among Brazilian isolates of C. fimbriata from other hosts. In the analyses of 14 microsatellite loci, the rubber tree isolates from Bahia were identical to each other and had alleles similar to those of Brazilian isolates from mango and eucalyptus. The microsatellite alleles of the Acre rubber tree isolates were identical to each other but distinct from other Brazilian isolates. The rubber tree isolates were morphologically indistinguishable from each other and very similar to the isolates of C. fimbriata on Ipomoea batatas, on which the species was originally described. Based on inoculation experiments results, the Bahia and Acre rubber tree isolates do not appear to be host specialized, which is typical for Brazilian isolates of C. fimbriata sensu stricto.

Teratosphaeria pseudoeucalypti on eucalyptus in Brazil

A new foliar disease caused by Teratosphaeria pseudoeucalypti on eucalyptus (E. globulus, E. urophylla x E. globulus and E. nitens x E. globulus) in Brazil is described . The disease is characterized by leaf spots of variable sizes and shapes, resulting in leaf blight and premature defoliation. Based on the morphological characteristics and multilocus phylogenetic analysis of the EF-1α, β-T and ITS-2 gene regions of five isolates, the fungus was identified as T. pseudoeucalypti. This is the first report of this pathogen outside Australia and a method for sporulation in culture is described.

Favorable conditions for Xanthomonas axonopodis infection in Eucalyptus spp.

Currently, Xanthomonas axonopodis is one of the main foliar pathogens for Eucalyptus spp. in Brazil. It induces leaf blight and defoliation of seedlings in the nursery and young plants in the field. However, little is known about thefavorable conditions for infection. The establishment and development of bacterial leaf blight caused by X. axonopodis in eucalyptus was studied for different leaf ages, temperatures and leaf wetness durations. Disease severity increased with leaf age, and the highest severity was observed on the fourth pair of completely expanded leaves (from the apex to the base). A higher level of bacterial colonization was also observed on the fourth pair of leaves quantified as bacterial cells/cm2 of leaf area. Twelve hours of free water on the leaf surface, prior to inoculation, was essential to promote a severe infection. However, with the increase in the leaf wetness duration, a decrease in disease severity was observed. The optimal temperature for disease development was 26-30oC.

Xanthomonas axonopodis pv. eucalyptorum pv. nov. Causing Bacterial Leaf Blight on Eucalypt in Brazil

Bacterial leaf blight is a major disease of eucalypt, especially under nursery conditions. Different bacterial species have been associated with the disease in several countries, and despite its importance worldwide, it is not clear to date whether similar disease symptoms are caused by the same or by different etiological agents. In this study, 43 bacterial strains were isolated from blighted eucalypt leaves collected in different geographic areas of Brazil and inoculated onto a susceptible eucalypt clone. Polyphasic taxonomy, including morphological, physiological, biochemical, molecular, and pathogenicity tests showed that only certain strains of Xanthomonas axonopodis caused symptoms of the disease. Strains varied in their aggressiveness, but no correlation with geographic origin was observed. MLSA-based phylogenetic analysis using concatenated dnaK, fyuA, gyrB and rpoD gene sequences allocated the strains in a well-defined clade, corresponding to Rade-marker’s group RG 9.6. Inoculation of nineteen plant species belonging to seven botanical families with representative strain LPF 602 showed it to be pathogenic only on Eucalyptus spp, and Corymbia spp. Based on distinct biochemical and pathogenic characteristics that differentiate the eucalypt strains from other pathovars of the X. axonopodis species, here we propose their allocation into the new pathovar X. axonopodis pv. eucalyptorum pv. nov.