48 results on '"Guimarães, Lúcio"'
Search Results
2. Early selection for rust resistance in eucalypt progenies and implications for other traits of interest in tree breeding.
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Arriel, Daniele Aparecida Alvarenga, Guimarães, Lúcio Mauro da Silva, Mafia, Reginaldo Gonçalves, Zauza, Edival Ângelo Valverde, and Alfenas, Acelino Couto
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EUCALYPTUS grandis ,TREE breeding ,WOOD density ,PHENOTYPES ,MOLECULAR cloning ,EUCALYPTUS - Abstract
Planting of resistant clones is the main control strategy for rust caused by Austropuccinia psidii on eucalypts in Brazil. Phenotyping for resistance to rust is performed at the last stage of the breeding programme on genetic material preselected for growth and industrial traits, which therefore may include susceptible genotypes. An alternative way to increase the frequency of resistant individuals in the breeding population is to perform resistance phenotyping during the initial stages of the programme. However, the impact of this approach on other characteristics considered as priority in the selection of genetic material should be assessed. In this study, we evaluated whether early selection for resistance to rust influenced total diameter at breast height (DBH), height (TH), average annual increment (IMA) and basic wood density (BWD), which are important selection criteria in eucalypt breeding. For this, a total of 6 703 plants from 70 full-sib progenies of Eucalyptus grandis, E. urophylla and hybrids of these species were phenotyped for rust resistance before transplanting, under controlled conditions, and again at seven months after transplanting, under field conditions and natural infection. Subsequently, the breeding population was evaluated for BWD indirectly through pilodyn penetration at 24 months after planting, and for DBH, TH and MAI at 37 months. Considering the final phenotype, 25.3% individuals were classified as resistant and 74.7% were susceptible to rust. Comparative genetic analyses between resistant and susceptible individuals showed that selection for resistance, performed early, did not influence DBH, TH, MAI or BWD traits. Thus, the early selection of resistant genotypes speeds up the exclusion of susceptible genotypes, saving time and resources in the development of new commercial clones without interfering with other important traits for tree breeding. [ABSTRACT FROM AUTHOR]
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- 2024
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3. Genetically differentiated populations of Ceratocystis fimbriata species complex points to host specialization in Brazil.
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Fernandes, Fernando Montezano, Azevedo, Daiana Maria Queiroz, da Silva Guimarães, Lúcio Mauro, Oliveira, Leonardo Sarno Soares, Alfenas, Rafael Ferreira, Júnior, Jaime Honorato, and Alfenas, Acelino Couto
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FIG ,GENETIC variation ,CACAO ,SPANNING trees ,PLANT species - Abstract
Ceratocystis fimbriata is an important pathogen with a wide host range that causes xylem lesions, wilt and death in different plant species. More adapted populations of C. fimbriata have probably been selected to specific hosts and high genetic and physiological variability can be found within the pathogen populations. Thus, molecular analysis and inoculation studies were performed to investigate the genetic and physiological variability of isolates of C. fimbriata complex obtained from different host species and geographic regions in Brazil. The minimum spanning tree analysis based on 14 simple‐sequence repeat (SSR) markers of C. fimbriata exhibited a clear clustering of isolates according to host, where each groups of isolates differed in at least five loci. In addition, inoculation of eight hosts with 10 isolates revealed a wide variation in aggressiveness. By assessing the length of xylem lesions caused by C. fimbriata isolates, Ficus carica was found to be the most susceptible host, followed by Mangifera indica. Only the isolates from Theobroma cacao and Carapa guianensis proved to be host specialized. Overall, the isolates tested were more aggressive to the hosts from which they were sampled. [ABSTRACT FROM AUTHOR]
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- 2024
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4. Correction to: Sensitive detection, quantification, and monitoring of Erwinia psidii colonization of guava plants using intercalating dye-based real-time PCR
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da Silva Hermenegildo, Pollyane, de Freitas, Rodrigo Galvão, de Souza Cascardo, Renan, Guimarães, Lúcio Mauro Silva, Badel, Jorge Luis, Alfenas‑Zerbini, Poliane, Marques, Abi S. A., Alfenas, Acelino Couto, and Ferreira, Marisa A. S. V.
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- 2023
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5. Transcriptome analysis of Eucalyptus grandis genotypes reveals constitutive overexpression of genes related to rust (Austropuccinia psidii) resistance
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Santos, Samuel A., Vidigal, Pedro M. P., Guimarães, Lúcio M. S., Mafia, Reginaldo G., Templeton, Matthew D., and Alfenas, Acelino C.
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- 2020
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6. Xanthomonas species causing leaf blight on eucalypt plants in Brazil and transfer of Xanthomonas axonopodis pv. eucalyptorum to Xanthomonas citri pv. eucalyptorum comb. nov.
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Ferraz, Hélvio Gledson Maciel, Badel, Jorge Luis, Neves, Yane Fernandes, Eloi, Ana Carolina Lopes, Vidigal, Pedro Marcus Pereira, Guimarães, Lúcio Mauro da Silva, and Alfenas, Acelino Couto
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XANTHOMONAS campestris ,EUCALYPTUS ,XANTHOMONAS ,COMMON bean ,SPECIES ,GENETIC variation - Abstract
Outbreaks of bacterial leaf blight (BLB) frequently affect eucalypt plants under nursery and field conditions in several countries. Although research has been conducted to unveil the causal agent, different bacterial species have been associated with similar disease symptoms in different countries. In order to determine the causal agent of BLB in Brazil, a survey was conducted in nine states to recover bacterial isolates from eucalypt plants exhibiting typical BLB symptoms. A total of 41 yellow‐colony isolates with varying aggressiveness towards a susceptible eucalypt clone were obtained, with 16S rDNA sequences indicating that they belong to the Xanthomonas genus. Rep‐PCR analysis separated the Xanthomonas population affecting eucalypt into six distinct groups revealing its high genetic diversity. The same population formed three clusters together with reference strains of X. citri, X. euvesicatoria and X. phaseoli in a phylogenetic tree constructed with partial dnaK, fyuA, gyrB and rpoD gene sequences. Clustering in the phylogenetic tree was clearly related to grouping based on rep‐PCR. Genome sequence comparisons of representative eucalypt isolates with type strains of validly published Xanthomonas species confirmed that the population consisted of X. citri, X. euvesicatoria and X. phaseoli. Inoculation of tomato, common bean, castor bean and eucalypt plants showed that the representative eucalypt isolates can cause disease in these plant species. Based on the results, the transfer of Xanthomonas axonopodis pv. eucalyptorum to Xanthomonas citri is proposed. These results are relevant for eucalypt BLB management under nursery and field conditions, including selection and deployment of effective plant resistance. [ABSTRACT FROM AUTHOR]
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- 2024
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7. Root infection and aerial colonization of eucalypt host plants by Erwinia psidii
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Montoya-Estrada, Claudia N., Costa, Camila R., Badel, Jorge L., Guimarães, Lúcio M. S., and Alfenas, Acelino C.
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- 2019
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8. Eucalypt powdery mildew caused by Podosphaera pannosa in Brazil
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Fonseca, Natália R., Guimarães, Lúcio M. S., Pires, Raul P., Klopfenstein, Ned B., and Alfenas, Acelino C.
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- 2017
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9. Ceratocystis fimbriata isolates on Mangifera indica have different levels of aggressiveness
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Oliveira, Leonardo S. S., Damacena, Michelle B., Guimarães, Lúcio M. S., Siqueira, Dalmo L., and Alfenas, Acelino C.
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- 2016
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10. QTL mapping for resistance to Ceratocystis wilt in Eucalyptus
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Rosado, Carla Cristina Gonçalves, da Silva Guimarães, Lúcio Mauro, Faria, Danielle Assis, de Resende, Marcos Deon Vilela, Cruz, Cosme Damião, Grattapaglia, Dario, and Alfenas, Acelino Couto
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- 2016
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11. An efficient inoculation method of Ralstonia solanacearum to test wilt resistance in Eucalyptus spp.
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Fonseca, Natália R., Oliveira, Leonardo S. S., Guimarães, Lúcio M. S., Teixeira, Ramon U., Lopes, Carlos A., and Alfenas, Acelino C.
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- 2016
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12. Outbreak of shoot blight and dieback of Eucalyptus spp., caused by Pseudoplagiostoma eucalypti in Brazil.
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Zauza, Edival Ângelo Valverde, da Silva Guimarães, Lúcio Mauro, Sales, Nilza de Lima Pereira, dos Santos, Samuel Alves, Alfenas, Rafael Ferreira, and Alfenas, Acelino Couto
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DIEBACK , *EUCALYPTUS , *LEAF spots , *SYMPTOMS , *DEFOLIATION , *ASCOMYCETES - Abstract
An outbreak of a new and severe disease was observed in Eucalyptus plantations of Bahia state, Brazil. An Ascomycota fungus has been frequently associated with the main symptoms of the disease namely leaf spot, branch cankers, shoot blight, defoliation, and dieback. Based on morphological characteristics, phylogenetic analysis (ITS and TEF‐1α genes), and pathogenicity test on Eucalyptus plants, Pseudoplagiostoma eucalypti was identified as the causal agent of the disease. Although P. eucalytpi has been known from in Brazil since 1998, this is the first report of it causing severe disease and die‐back on Eucalyptus spp. and we also record new symptoms associated with the pathogen. [ABSTRACT FROM AUTHOR]
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- 2023
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13. Infection process of Puccinia psidii in Eucalyptus grandis leaves of different ages
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Xavier, Adelica Aparecida, da Silva, André Costa, da Silva Guimarães, Lúcio Mauro, Matsuoka, Kiyoshi, Hodges, Charles S., and Alfenas, Acelino Couto
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- 2015
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14. Genetic variation, morphology and pathogenicity of Ceratocystis fimbriata on Hevea brasiliensis in Brazil
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Valdetaro, Denise C. O. F., Oliveira, Leonardo S. S., Guimarães, Lúcio M. S., Harrington, Thomas C., Ferreira, Maria A., Freitas, Rodrigo G., and Alfenas, Acelino C.
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- 2015
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15. Mycosphaerella and Teratosphaeria species associated with leaf diseases on Eucalyptus globulus in southern Brazil
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TEODORO, Marcela G., FERREIRA, Maria A., GUIMARÃES, Lucio M.S., MAFIA, Reginaldo G., GROENEWALD, Johannes Z., CROUS, Pedro W., and ALFENAS, Acelino C.
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- 2012
16. Genetic mapping and validation of QTLs associated with resistance to Calonectria leaf blight caused by Calonectria pteridis in Eucalyptus
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Zarpelon, Talyta Gafassi, da Silva Guimarães, Lúcio Mauro, Faria, Danielle Assis, Coutinho, Marcelo Magalhães, Cápua Neto, Braz, Teixeira, Ramon Ubirajara, Grattapaglia, Dario, and Alfenas, Acelino Couto
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- 2015
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17. 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
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Alves, Alexandre Alonso, Rosado, Carla Cristina Gonçalves, Faria, Danielle Assis, Guimarães, Lúcio Mauro da Silva, Lau, Douglas, Brommonschenkel, Sérgio Hermínio, Grattapaglia, Dario, and Alfenas, Acelino Couto
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- 2012
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18. Leaf gas exchange and chlorophyll a fluorescence of Eucalyptus urophylla in response to Puccinia psidii infection
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Alves, Alexandre Alonso, Guimarães, Lúcio Mauro da Silva, Chaves, Agnaldo Rodrigues de Melo, DaMatta, Fábio Murilo, and Alfenas, Acelino Couto
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- 2011
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19. A new race of Puccinia psidii defeats rust resistance in eucalypt
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Graça, Rodrigo N., Aun, Cristina P., Guimarães, Lúcio M. S., Rodrigues, Bruno V. A., Zauza, Edival A. V., and Alfenas, Acelino C.
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- 2011
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20. QTL associated with resistance to defoliation (Cylindrocladium pteridis) in Eucalyptus spp.
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Zarpelon Talyta G, Guimarães Lúcio MS, Coutinho Marcelo M, Neto Braz Cápua, Faria Danielle A, Grattapaglia Dario, and Alfenas Acelino C
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Medicine ,Science - Published
- 2011
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21. Validation and use of a qPCR protocol to quantify the spread of Ralstonia solanacearum in susceptible and resistant eucalypt plants.
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Freitas, Rodrigo G., Hermenegildo, Pollyane S., Cascardo, Renan S., Guimarães, Lúcio M. S., Santos, Samuel A., Badel, Jorge L., Alfenas‐Zerbini, Poliane, and Alfenas, Acelino C.
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RALSTONIA solanacearum ,EUCALYPTUS ,BACTERIAL DNA ,WILT diseases ,PHENOTYPES ,PLANT cells & tissues ,DISEASE management - Abstract
Bacterial wilt caused by Ralstonia solanacearum is a serious disease of eucalypt in humid and high temperature areas worldwide. Spreading of the bacterium in the field or to other nurseries occurs mainly by symptomless infected plant material. The use of pathogen‐free propagating material as well as planting of resistant genotypes are currently the only strategies used for disease control. Therefore, a reliable and sensitive method for detection of low titres of R. solanacearum in infected plant tissue is essential for the success of management programmes. In this work, we adapted an efficient intercalating dye‐based real‐time PCR protocol to detect the bacterium in symptomless eucalypt plants as well as to investigate its movement in eucalypt clones CLR172 and CLR371, which exhibit resistant and susceptible phenotypes, respectively. We found that the bacterium translocates acropetally and basipetally in inoculated but symptomless cuttings of the resistant clone, as in cuttings of the susceptible clone displaying symptoms. Nevertheless, a smaller concentration of bacterial DNA was detected in tissues of the resistant clone. Mature biofilms occluding the xylem vessels were present in the susceptible clone whereas only single cells or small aggregates were observed in the resistant clone. This work contributes to improve our knowledge of the colonization process of R. solanacearum in eucalypt clones with different levels of susceptibility and to understand how the defence mechanisms against bacterial wilt in Eucalyptus work. Our findings could aid in the selection of the most resistant eucalypt clones to be used in wilt disease management programmes. [ABSTRACT FROM AUTHOR]
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- 2021
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22. Molecular characterization and aggressiveness of the Ralstonia solanacearum species complex from Eucalyptus spp. in Brazil.
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Freitas, Rodrigo G., Hermenegildo, Pollyane S., Guimarães, Lúcio M. S., Zauza, Edival A. V., Badel, Jorge L., and Alfenas, Acelino C.
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RALSTONIA solanacearum ,EUCALYPTUS ,DNA fingerprinting ,SPECIES ,DISEASE resistance of plants ,RALSTONIA - Abstract
Bacterial wilt, one of the world's most destructive diseases in many crops, including eucalypt, is caused by four distinct phylogenetic lineages of the Ralstonia solanacearum species complex, recently classified in three distinct species: R. solanacearum (phylotype II), R. pseudosolanacearum (phylotype I and III) and R. syzygii (phylotype IV). In this study, we characterized 93 Ralstonia isolates obtained from eucalypt grown in different Brazilian regions using phylotype and sequevar designations and genomic fingerprinting with BOX‐PCR. In addition, we evaluated the aggressiveness of a select group of isolates in two eucalypt clones differing in resistance. We found that 89 isolates belong to R. solanacearum (phylotype II) and four to R. pseudosolanacearum (phylotype I). Unlike R. pseudosolanacearum, R. solanacearum was phylogenetically diverse and no correlation was found between sequevar and geographic origin. Most isolates grouped with reference isolates of phylotype IIA sequevar 41, whereas a few others clustered in phylotype IIB, mainly sequevar 4NPB, which is an emergent variant described affecting eucalypt for the first time. Isolates of R. solanacearum phylotype IIB were less aggressive to clone CLR371 (susceptible) whereas the R. pseudosolanacearum isolate tested was the only one pathogenic to the CLR172 (resistant) clone. Isolate aggressiveness varied between the eucalypt clones tested. The results of this study reinforce the importance of conducting molecular and aggressiveness characterization of the pathogen population to develop management strategies aimed at the deployment of host resistance in eucalypt breeding programmes. [ABSTRACT FROM AUTHOR]
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- 2021
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23. Improved evaluation of the genetic variability of Brazilian strains of Erwinia psidii with newly developed microsatellite markers.
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Hermenegildo, Pollyane da Silva, Guimarães, Lúcio Mauro Silva, Badel, Jorge Luis, Marques, Abi S. A., Alfenas, Acelino Couto, and Velloso Ferreira, Marisa A. S.
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MICROSATELLITE repeats , *ERWINIA , *GUAVA , *TANDEM repeats , *EUCALYPTUS , *SPANNING trees , *SHORT tandem repeat analysis , *DIEBACK - Abstract
Erwinia psidii is the causal agent of bacterial blight of guava, and wilt and dieback of eucalypt in Brazil. Previous studies using repetitive sequence‐based PCR (rep‐PCR) revealed limited genetic diversity in E. psidii populations. Here, the draft genome sequence of the type strain IBSBF435 was probed for variable number of tandem repeats (VNTRs) that could be used to study E. psidii genetic diversity by multilocus variable number of tandem repeat analysis (MLVA). Eleven selected VNTRs were used to assess genetic relatedness among 108 E. psidii strains from guava (36) and eucalypt (72), of different geographic origins and years of collection. A total of 79 haplotypes were detected in the strain collection. The number of alleles per locus ranged from 3 to 15. A minimum spanning tree indicated the occurrence of nine clonal complexes, of which two contained only guava strains, five only eucalypt strains, and two contained guava and eucalypt strains. The three oldest strains collected from guava in the 1980s (IBSBF435, IBSBF454, and IBSBF493) exhibited distinct haplotypes. IBSBF435 was genetically more closely related to some eucalypt strains than to guava strains, which supports previously reported lack of host specificity. IBSBF454 grouped with guava strains more recently collected from different states, suggesting that it could have been disseminated to different Brazilian areas. The implication of the lack of differentiation between guava and eucalypt strains is discussed in relation to the detection of E. psidii affecting eucalypt more than 25 years after the first description of guava bacterial blight. [ABSTRACT FROM AUTHOR]
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- 2021
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24. Prediction, structure characterization, and evolutionary analysis of Erwinia psidii putative type III effectors.
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Pereira, Isadora C., Badel, Jorge L., Vidigal, Pedro M. P., Sousa, Adryelle A., Santos, Samuel A., Guimarães, Lúcio M. S., and Alfenas, Acelino C.
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EUCALYPTUS ,ERWINIA ,HORIZONTAL gene transfer ,ERWINIA amylovora ,GUAVA ,GRAM-negative bacteria ,GENES - Abstract
Erwinia psidii is a gram‐negative bacterium that threatens both guava and eucalypt plantations in several countries. Despite the economic importance of both crops, nothing is currently known about the molecular mechanisms underlying E. psidii pathogenicity and, consequently, how it evolved to infect Eucalyptus species besides its presumed native host Psidium guajava. In this study, we predicted putative type III secretion system effectors that may play important roles during plant–E. psidii interactions and conducted effector structure and phylogenetic analyses to gain important insights into their function and evolution. For that, the whole genomes of four E. psidii strains that exhibit differential aggressiveness towards eucalypt clones were sequenced and their effector repertoires predicted based on sequence identity with known effectors of the model phytopathogen Erwinia amylovora. Only proteins sharing significant sequence identity with the DspE and Eop1 effectors were found. Here, it is shown that these two E. psidii effectors retain all structural characteristics of their corresponding protein superfamilies, but exhibit allelic variations that are consistent with the observed aggressiveness differences between strains. Phylogenetic analyses revealed that whereas E. psidii housekeeping gene sequences are more closely related to those from Erwinia tracheiphila, the effector (either nucleotide or amino acid) sequences are more closely related to their Pantoea agglomerans counterparts, suggesting that dspE and eop1 were both acquired through horizontal gene transfer from the latter bacterial species. The results of this study provide important insights on E. psidii pathogenicity and set the stage for future effector functional studies. [ABSTRACT FROM AUTHOR]
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- 2021
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25. A new, highly aggressive race of Austropuccinia psidii infects a widely planted, myrtle rust‐resistant, eucalypt genotype in Brazil.
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Almeida, Rosiane F., Machado, Patrícia S., Damacena, Michelle B., Santos, Samuel A., Guimarães, Lúcio M.S., Klopfenstein, Ned B., and Alfenas, Acelino C.
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EUCALYPTUS ,GENOTYPES ,GENES ,PLANT selection - Abstract
Myrtle rust (MR) caused by Austropuccinia psidii is one of the most important diseases affecting eucalypt (Eucalyptus) plantations in Brazil. Over the years, selection and planting of MR‐resistant clones has been the primary strategy for MR management. In May 2013, young trees of the GG100 hybrid (E. grandis × E. urophylla) clone—widely planted in Brazil and previously classified as resistant to MR—were infected by A. psidii in Minas Gerais, Brazil. In this study, artificial inoculations of a eucalypt clone set with differential reactions to A. psidii races were used to discover a new race of A. psidii (race 5) that was highly aggressive on the majority of eucalypt clones tested. In addition, only this new race successfully infected eucalypt G26 and 847 genotypes, which were formerly classified as resistant to the four previously known races of A. psidii. Since G26 genotype is homozygous for Ppr1 (a major resistance gene against A. psidii), this is the first report of MR resistance breakdown in a eucalypt genotype homozygous for Ppr1. Our findings demonstrate that this new A. psidii race is highly aggressive and capable of infecting a larger number of eucalypt genotypes compared with the previously known A. psidii races 1, 2, 3, and 4. [ABSTRACT FROM AUTHOR]
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- 2021
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26. Resistance of kiwifruit cultivars to ceratocystis wilt: An approach considering the genetic diversity and variation in aggressiveness of the pathogen.
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Oliveira, Leonardo S. S., Pimenta, Lucas V. A., Guimarães, Lúcio M. S., Souza, Paulo V. D., Bhering, Leonardo L., and Alfenas, Acelino C.
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KIWIFRUIT ,CULTIVARS ,WILT diseases ,ACTINIDIA ,ROOTSTOCKS ,PATHOGENIC microorganisms - Abstract
Kiwifruit (Actinidia spp.) is native to southern China, but was first cultivated in New Zealand and then spread worldwide. Emerging diseases such as ceratocystis wilt have attracted the attention of kiwifruit growers due to the great losses observed in southern Brazil. Effective control can be achieved by screening for resistance, but the genetic variability of the pathogen must be considered. Thus, this study aimed to assess the genetic diversity and variation in aggressiveness of Ceratocystis isolates from kiwifruit in southern Brazil and then evaluate the resistance of kiwifruit cultivars with the most aggressive isolates. A collection of 46 isolates were obtained from southern Brazil and 14 simple‐sequence repeat (SSR) markers was successfully used for genotyping. Out of 14 markers, 13 were polymorphic and identified 26 genotypes. Fourteen distinct genotypes were tested on a susceptible cultivar to select the most aggressive ones. Finally, inoculation with an equal mixture of five of the most aggressive isolates was used to evaluate the resistance of seven kiwifruit cultivars: Red Arguta, Green Arguta, Allison, Chieftain, Hayward, Monty, and Tomury. Cultivars varied in levels of susceptibility, with disease severity ranging from 40% to 100%. Considering the length of stem lesions, Chieftain showed the lowest level of severity at 40%, while no wilt symptoms were observed at 45 days after inoculation. In addition to the seven cultivars, a half‐sibling progeny with 618 plants of the rootstock cv. Bruno was also assessed, but only seven individuals were resistant. These seven plants can be cloned and used as resistant rootstocks in commercial orchards. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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27. Detection and characterization of Ralstonia pseudosolanacearum infecting Eucalyptus sp. in Brazil.
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Freitas, Rodrigo G., Hermenegildo, Pollyane S., Guimarães, Lúcio M. S., Zauza, Edival A. V., Badel, Jorge L., Alfenas, Acelino C., and Hietala, A. M.
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EUCALYPTUS ,RALSTONIA ,RALSTONIA solanacearum ,PHYTOPATHOGENIC bacteria ,NUCLEOTIDE sequence ,TREE crops - Abstract
Ralstonia solanacearum sensu lato causes bacterial wilt in many agronomic crops and tree species economically important worldwide. It is a species complex that has been divided into phylotypes and sequevars, commonly related to geographic distribution. Knowledge of the phylotype composition and genetic variability in populations of this phytopathogenic bacterium is useful for implementing effective control measures. In a survey conducted in 2019, six bacterial strains were obtained from wilted Eucalyptus urophylla trees in plantations located in the municipality of Dom Eliseu, Pará state, Brazil. Multiplex PCR based on the internal transcribed spacer (ITS) indicated that the bacterial strains belonged to two different species, namely R. pseudosolanacearum (phylotype I) and R. solanacearum (phylotype II). In a phylogenetic analysis, the nucleotide sequence of the endoglucanase (egl) gene from eucalypt strains of phylotype I clustered together with sequevar 18 sequences from GenBank. Separation of the strains into two different species was confirmed by repetitive element palindromic PCR (rep‐PCR). Pathogenicity tests demonstrated that the R. solanacearum and R. pseudosolanacearum strains recovered from E. urophylla cause disease in both tomato and eucalypt plants. Until now, only R. solanacearum (Phylotype II) has been reported causing wilt symptoms on Eucalyptus spp. in Brazil. Therefore, the presence of R. pseudosolanacearum and a need for better understanding of its genetic and aggressiveness variability as well as possible differences between the two species should be considered in breeding programmes aimed at the deployment of host resistance. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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28. Resistance of Eucalyptus pellita to rust (Puccinia psidii)
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Santos, Marisângela Rodrigues, Guimarães, Lúcio Mauro da Silva, Resende, Marcos Deon Vilela de, Rosse, Leonardo Novaes, Zamprogno, Karina Carnielli, and Alfenas, Acelino Couto
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doença ,disease ,Resistência genética ,herdabilidade ,quantitative inheritance ,Genetic resistance ,heritability ,herança quantitativa - Abstract
Eucalypts rust (Puccinia psidii) is currently one of the major diseases in commercial eucalypt plantations in Brazil. The primary method of disease control is the use of resistant genotypes, and, among the different species of Eucalyptus, E. pellita is indicated as a promising source of resistance. In this work, the genetic control of rust resistance in E. pellita through inoculations under controlled conditions of 441 plants from four full-sibling families was studied. Inoculations were performed using the monopostular isolate UFV-2, race 1. All families tested segregated for rust resistance, and the number of resistant plants was higher than susceptible in all crosses. Inheritance models based on few genes did not fully explain the observed segregation patterns, and the narrow-sense heritability of rust resistance was estimated between 32.7% and 37.3%. The results suggested that rust resistance in E. pellita is complex and is controlled by major- and minor-effect genes. A ferrugem do eucalipto (Puccinia psidii) é atualmente uma das principais enfermidades em plantios comerciais de eucalipto no Brasil. Dentre as diferentes espécies de eucalipto, Eucalyptus pellita é apontada como uma promissora fonte de resistência. Neste trabalho estudou-se o controle genético da resistência à ferrugem em E. pellita por meio de inoculações em condições controladas de 441 plantas oriundas de quatro progênies. As inoculações foram realizadas com o isolado monopostular UFV-2, raça 1. Todas as progênies segregaram para resistência à ferrugem, sendo o número de plantas resistentes superior em todos os cruzamentos. Modelos de herança baseados em poucos genes não explicaram totalmente os padrões de segregação obtidos. A herdabilidade no sentido restrito da resistência à ferrugem foi estimada entre 32,7% a 37,3%. Os resultados obtidos sugerem que a resistência à ferrugem em E. pellita é complexa, sendo governada por genes de efeito maior e menor.
- Published
- 2014
29. Genetic control of resistance on Mangifera indica to Ceratocystis wilt.
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Arriel, Daniele Aparecida Alvarenga, Guimarães, Lúcio Mauro da Silva, Resende, Marcos Deon Vilela de, Lima Neto, Francisco Pinheiro, Silva, Daniella Flávia Said Heid Schettini, Siqueira, Dalmo Lopes de, and Alfenas, Acelino Couto
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MANGO diseases , *CERATOCYSTIS diseases , *DISEASE resistance of plants , *ALLELES in plants ,FRUIT genetics - Abstract
Ceratocystis wilt, caused by Ceratocystis fimbriata is one of the most serious limiting factors for mango production in Brazil. Despite efforts in the selection and the identification of mango cultivars resistant to Ceratocystis wilt, the genetic basis of the resistance remains unknown. Therefore, the objective of this study was to understand the inheritance of resistance to C. fimbriata by artificial inoculations of the pathogen in progenies of six commercial varieties of mango using “Tommy Atkins” as the male parent. The cultivars “Keitt”, “Palmer”, “Tommy Atkins” and “Van Dyke” were confirmed as moderately resistant, whereas “Coquinho”, “Espada” and “Haden” were susceptible. The results of the inoculation on the progenies of these cultivars revealed that the resistance in mango is polygenic with a prevalence of genes expressing the effects of dominance and epistasis. The genetic gain with the selection of the 10 more resistant plants was 46%, which indicated a 46% reduction in disease severity. In general, a low frequency of the alleles favorable to disease resistance was observed in the population studied, which suggests the need for the introduction of new sources of genetic materials carrying the genes responsible for resistance. [ABSTRACT FROM AUTHOR]
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- 2016
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30. Eucalyptus pellita as a source of resistance to rust, ceratocystis wilt and leaf blight.
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Guimarães, Lúcio Mauro da Silva, Titon, Miranda, Lau, Douglas, Rosse, Leonardo Novaes, Oliveira, Leonardo Samo Soares, Rosado, Carla Cristina Gonçalves, Christo, Guilherme Gegenheiner Ornelas, and Alfenas, Acelino Couto
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PLANT breeding , *EUCALYPTUS , *CERATOCYSTIS , *TARO leaf blight , *CYLINDROCLADIUM , *GENETIC polymorphisms , *PLANT diseases , *PUCCINIA , *DISEASE resistance of plants - Abstract
Rust (Puccinia psidii), ceratocystis wilt (Ceratocystis fimbriata) and cylindrocladium leaf blight (Cylindrocladium pteridis) are important diseases of eucalyptus. Planting of resistant genotypes is the most suitable control strategy of forest diseases under field condition. Resistance level of 23 Eucalyptus pellita clones was evaluated by artificial inoculations. Among the inoculated clones, 12 were resistant to rust, 16 to ceratocystis wilt and 12 to cylindrocladium leaf blight, and three of them were resistant to all three diseases. The high intra-specific variability found in this study demonstrates the importance of E. pellita as a disease resistance source to be employed for introgression of novel resistance genes in eucalyptus genetic breeding programs. [ABSTRACT FROM AUTHOR]
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- 2010
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31. Resistance of mango cultivar Ubá to Ceratocystis fimbriata depends on the pathogen's physiological variability.
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Guimarães, Lúcio M.S., Nunes, Angélica S., Santos, Samuel A., Resende, Marcos D.V., Damacena, Michelle B., Siqueira, Dalmo L., Alves, Rodrigo S., and Alfenas, Acelino C.
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MANGO ,GRAFTING (Horticulture) ,ROOTSTOCKS ,VACCINATION - Abstract
Mango sudden decline (MSD) caused by Ceratocystis fimbriata is one of the major diseases affecting mango crop in Brazil, Oman, and Pakistan. In Brazil, the main strategy for MSD management has been the use of resistant plants as rootstock for grafting. In this study, we tested the resistance of mango cultivar Ubá to C. fimbriata isolates from different populations. Firstly, we inoculated 2173 seedlings from 129 accessions of mango cv. Ubá using three representative fungal isolates from Brazil. Following the inoculation, the potential resistant plants were challenged against ten C. fimbriata isolates from Brazil and three from Oman. Finally, to confirm its resistance, clonal replicates of the resistant accessions were re-inoculated with the same fungal isolates. In the first selection, 13 accessions were pre-selected as resistant. In subsequent selections, they proved to be resistant to C. fimbriata isolates from populations from Oman/Pakistan and Southeastern Brazil. However, all 13 accessions were susceptible to fungal isolates from populations from Northeastern Brazil and Rio de Janeiro (also in Brazil). Our results reveal that mango cv. Ubá resistance to MSD is isolate specific and depends on the pathogen population. We obtained 13 mango cv. Ubá clones resistant to C. fimbriata , which may be useful as rootstock to MSD management in Oman and specific regions from Brazil. • The resistance of mango cultivar Ubá to mango sudden decline (MSD) is isolate specific. • Thirteen clones of mango cv. Ubá resistant to MSD were obtained. • The resistant clones may be used as rootstock in commercial orchards of mango. [ABSTRACT FROM AUTHOR]
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- 2021
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32. An agglutination test for fast and specific detection of Erwinia psidii.
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Montoya‐Estrada, Claudia N., Silva, Patrick F., Costa, Camila R., Oliveira, Leandro L., Guimarães, Lúcio M.S., Badel, Jorge L., Alfenas, Acelino C., and Desprez‐Loustau, Marie‐Laure
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AGGLUTINATION tests ,GENE amplification ,ERWINIA ,PATHOGENIC bacteria ,GUAVA ,PLANT cells & tissues ,POLYMERASE chain reaction ,IMMUNOGLOBULINS - Abstract
Dieback and wilt, caused by Erwinia psidii (Ep), is one of the most important emergent diseases of Eucalyptus spp. in Brazil. Currently, pathogen detection relies on isolation of bacteria from infected plant tissue and either identification based on morphological, physiological and biochemical tests or DNA amplification using the polymerase chain reaction (PCR), which in many cases is laborious and cumbersome. Considering the need for a simpler and more rapid, yet reliable, method for detecting the pathogen, we obtained a polyclonal antibody (anti‐Ep) and developed an agglutination test for specific detection of E. psidii. The antiserum was produced against the E. psidii strain LPF534 and tested against 101 E. psidii isolates from Eucalyptus spp.; three E. psidii isolates from Psidium guajava; 23 Ralstonia solanacearum and 18 Xanthomonas axonopodis isolates pathogenic to Eucalyptus spp.; and seven endophytic isolates from Eucalyptus spp., three of which are phylogenetically related to the genus Erwinia. Results of direct ELISA indicated that a concentration as low as 3.5 µg/ml of the anti‐Ep antibody was able to detect the E. psidii antigen and that the antibody did not cross‐reacted with other bacteria pathogenic and non‐pathogenic to Eucalyptus spp. In the agglutination test, the anti‐Ep antibody showed positive reaction with all strains of E. psidii tested whereas cross‐reaction with none of the strains that belong to other taxonomic groups was observed. The agglutination test showed a detection limit of 105 colony‐forming units (CFU)/ml, and its specificity was the same as that obtained by PCR amplification using E. psidii‐specific primers. These results demonstrate that the agglutination test developed here is a useful tool for specific, fast and inexpensive detection of E. psidii although only operational on pure bacterial suspensions and not yet directly from infected tissues. [ABSTRACT FROM AUTHOR]
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- 2019
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33. Host range of Erwinia psidii and genetic resistance of Eucalyptus and Corymbia species to this pathogen.
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Caires, Nilmara P., Hermenegildo, Pollyane S., Guimarães, Lúcio M. S., Mafia, Reginaldo G., Zauza, Edival Â. V., Júnior, Norton Borges, Badel, Jorge L., and Alfenas, Acelino C.
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EUCALYPTUS ,SPECIES ,ERWINIA ,PLANT species ,DISEASE resistance of plants ,INFORMATION resources ,PATHOGENIC microorganisms - Abstract
Dieback caused by Erwinia psidii is currently one of the most important emerging diseases in eucalypt plantations in Brazil. However, little is known in terms of the host range of this pathogen or the potential sources of resistance against the disease it causes. In this study, we inoculated plants of species from nine families to gain insight into the host range of E. psidii. Plants of all inoculated species of Myrtaceae except Acca sellowiana exhibited disease symptoms and therefore represent potential hosts for the pathogen under natural conditions. In addition, the response of four Corymbia species, 29 Eucalyptus species and three interspecific Eucalyptus hybrids to inoculation with E. psidii was evaluated. All Corymbia henryi, Corymbia maculata, Eucalyptus thozetiana, Eucalyptus cloeziana, Eucalyptus viminalis, Eucalyptus dalrympleana and Eucalyptus pilularis plants were highly resistant to the pathogen, whereas differential disease resistance was observed in the other species. This study provides important information on sources of resistance to Erwinia psidii with potential use in the development of clones with enhanced resistance in eucalypt species of economic importance. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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34. Transcriptome analysis of a powdery mildew pathogen (Podosphaera pannosa) infecting Eucalyptus urophylla: De novo assembly, expression profiling and secretome prediction.
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Fonseca, Natália R., Ibarra Caballero, Jorge, Kim, Mee‐Sook, Stewart, Jane E., Guimarães, Lúcio M. S., Alfenas, Acelino C., Klopfenstein, Ned B., and Fossdal, Carl Gunnar
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EUCALYPTUS ,POWDERY mildew diseases ,TETRAHYDROFOLATE dehydrogenase ,RNA sequencing ,DISEASE resistance of plants ,FUNGAL growth - Abstract
Podosphaera pannosa is the causal agent of powdery mildew on eucalypt in Brazil. This powdery mildew disease is important in nurseries causing leaf and shoot distortion, shoot discoloration and reduction in growth, which decreases mini‐cutting production. Improved RNA sequencing (RNA‐Seq) technologies have allowed increased information about the transcriptome of several pathogens and hosts, enabling a better understanding of their interaction at the gene level. For this study, we analysed the transcriptome of P. pannosa during leaf infection of Eucalyptus urophylla leaves using RNA‐Seq and de novo transcriptome assembly. The transcriptome was Illumina sequenced and assembled de novo, generating over 178 million RNA‐Seq reads assembled onto 200,473 contigs. After filtering steps, the resulting 12,106 (6%) transcripts were identified as the P. pannosa transcriptome data set. The 10 most abundant transcripts included genes encoding enzymes likely involved in fungal establishment and growth, such as dihydrofolate reductase, putative methyltransferases, acyl‐desaturase, glycoside hydrolase and dehydrogenases. In addition, genes putatively encoding an aquaporin and an orthologue to the effector protein, GoEC2 of Golovinomyces orontii were identified. The predicted secretome consisted of 1,899 translated transcripts, of which 310 exhibited homology to proteins described in the PHI database, 144 of these showing homology to fungal PHI accessions that affected pathogenicity or that are described as effectors. In addition, 81 transcripts encoded secreted proteins homologous to effectors described in the Erysiphales. These results provide a basis for continued studies to better understand the P. pannosa‐eucalypt (Eucalyptus spp.) pathosystem and could parallel studies of the eucalypt transcriptome to help determine host resistance mechanisms. [ABSTRACT FROM AUTHOR]
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- 2019
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35. Diversity, prevalence and phylogenetic positioning of Botrytis species in Brazil.
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Azevedo, Daiana M.Q., Martins, Sarah D.S., Guterres, Débora C., Martins, Mateus D., Araújo, Leonardo, Guimarães, Lúcio M.S., Alfenas, Acelino C., and Furtado, Gleiber Q.
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BOTRYTIS , *RNA polymerase II , *EUCALYPTUS , *BOTRYTIS cinerea , *HOST plants - Abstract
Botrytis is a necrotrophic fungal genus of great economic importance worldwide. Together, the Botrytis species are able to infect over one thousand host plant species, including dicotyledons and monocotyledons. As the identification of Botrytis species in Brazil has mostly been based only on morphological characterization and comparisons of the rDNA ITS region, which is not informative in the genus, its diversity remains unknown. Thus, in this study we determined the diversity and prevalence of Botrytis spp. in Brazil by multilocus phylogeny. Phylogenetic reconstruction of the genus was performed using the nuclear genes glyceraldehyde-3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60) and RNA polymerase II second largest subunit (RPB2). From analyses of 56 Botrytis isolates obtained from different hosts and geographical regions in Brazil, we found that Botrytis cinerea is the most prevalent species with considerable intraspecific genetic diversity detected by nuclear genes. Two new hosts to B. cinerea and eight host never previously reported in Brazil were found. We also reported for the first time the occurrence of B otrytis pseudocinerea associated with A cca se l lowiana (Myrtaceae). Due to the new phylogenetic positioning of B otrytis pelargonii and B otrytis eucalypti , a taxonomic review of these species was suggested. • Botrytis cinerea is the prevalent Botrytis species in Brazil. • Botrytis pseudocinerea was reported in Brazil associated with Acca sellowiana (Myrtaceae). • Three new plant–pathogen combinations were described. • Eight new hosts to B. cinerea in Brazil were reported. [ABSTRACT FROM AUTHOR]
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- 2020
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36. A host specialized form of Ceratocystis fimbriata causes seed and seedling blight on native Carapa guianensis (andiroba) in Amazonian rainforests.
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Valdetaro, Denise C.O.F., Harrington, Thomas C., Oliveira, Leonardo S.S., Guimarães, Lúcio M.S., McNew, Douglas L., Pimenta, Lucas V.A., Gonçalves, Rivadalve C., Schurt, Daniel A., and Alfenas, Acelino C.
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CERATOCYSTIS , *SEEDLINGS , *CRABWOOD , *RAIN forests , *MICROSATELLITE repeats - Abstract
Abstract Ceratocystis fimbriata Ellis & Halsted recently was recorded causing seed and seedling blight on Carapa guianensis Aubl. (andiroba), a tree species native to the Amazon Rainforest and prized for its valuable timber and medicinal seed oil. C. fimbriata more commonly causes wilt type diseases in woody hosts, especially on non-native host trees. However, on andiroba the disease occurs on seedlings and seeds, affecting the species regeneration. We studied 73 isolates of C. fimbriata on andiroba from three regions of the Amazon Basin to see if they represented natural or introduced populations. Analysis of ITS rDNA sequences and phylogenetic analysis of mating type genes revealed new haplotypes of C. fimbriata from the Latin American Clade that were closely related to other Brazilian populations of the fungus. In mating experiments, andiroba isolates were inter-fertile with tester strains of C. fimbriata from Brazil and elsewhere, confirming that they belong to a single biological species. Using microsatellite markers, 14 genotypes and populations with intermediate levels of genetic variability were found, suggesting that the fungus is indigenous to the Amazon Basin. Inoculation tests indicated that the andiroba isolates are host-specialized on andiroba, supporting the proposition of the special form C. fimbriata f. sp. carapa. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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37. Quantificação, caracterização molecular e agressividade do complexo de espécies Ralstonia solanacearum em Eucalyptus spp
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Freitas, Rodrigo Galvão de, Guimarães, Lúcio Mauro da Silva, Pacheco, Jorge Luis Badel, Zerbini, Poliane Alfenas, and Alfenas, Acelino Couto
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Eucalipto - Genética ,Fitopatologia ,Reação em cadeia da polimerase em tempo real ,Murcha-bacteriana ,Eucalipto - Resistência a doenças e pragas - Abstract
A murcha bacteriana causada por Ralstonia solanacearum é uma importante doença do eucalipto em regiões quentes e úmidas em todo o mundo. A bactéria exibe considerável variação quanto à gama de hospedeiros, origem geográfica, patogenicidade e às propriedades fisiológicas. A dispersão da bactéria para o campo ou para outros viveiros de eucalipto ocorre principalmente por material vegetal infectado, porém assintomático. O uso de material de propagação livre do patógeno, bem como o plantio de genótipos resistentes são atualmente as únicas estratégias utilizadas para o controle da doença. Portanto, o conhecimento dos filotipos e da variabilidade genética em populações dessa bactéria é importante para a implementação de medidas de controle eficientes. Neste trabalho, desenvolveu-se um eficiente protocolo de PCR em tempo real baseado em corante intercalante para detectar a bactéria em plantas assintomáticas de eucalipto, bem como investigar sua movimentação nos tecidos de clones com diferentes níveis de resistência. Além disso, identificamos filotipos, sequevares e genótipos de 93 isolados de Ralstonia, obtidos de mudas e árvores de eucalipto cultivadas em diferentes regiões do Brasil. Posteriormente, foram investigados os padrões de distribuição de sequevar e sua relação com a agressividade da bactéria em clones de eucalipto. Através da técnica de PCR em tempo real, observou-se que a bactéria translocou acropetal e basipetalmente em plantas inoculadas e assintomáticas do clone resistente, assim como em plantas sintomáticas do clone suscetível. No entanto, uma menor concentração bacteriana foi detectada nos tecidos do clone resistente, onde, por meio de microscopia eletrônica de varredura, não se observou biofilme bacteriano obstruindo os vasos do xilema. Por meio de análises moleculares, constatou-se que, assim como R. solanacearum (filotipo II), R. pseudosolanacearum (filotipo I) também causa murcha em plantas de eucalipto. Entretanto, R. pseudosolanacearum é geneticamente uniforme, o que indica uma provável recente introdução desse filotipo no país. Diferentemente de R. pseudosolanacearum, R. solanacearum é filogeneticamente diversa, e não há correlação entre sequevar e origem geográfica. A agressividade dos isolados variou entre os clones de eucalipto testados, o que reforça a importância da caracterização molecular e de estudos de agressividade da população do patógeno, a fim de embasar a seleção de material resistente. O método de qPCR desenvolvido neste estudo pode ser valioso para detecção do patógeno durante o diagnóstico da doença, assim como para sua quantificação nos tecidos da planta. O presente estudo expande o conhecimento da variabilidade genética do complexo de espécies de Ralstonia solanacearum em Eucalyptus spp. Palavras-chave: Murcha bacteriana. PCR em Tempo Real. Eucalipto. Bacterial wilt caused by Ralstonia solanacearum is a serious disease of eucalypt in humid and high temperature areas worldwide. The bacterium exhibits considerable variation in the host range, geographic origin, pathogenicity and physiological properties. Spreading of the bacterium in the field or to other eucalypt nurseries occurs mainly by infected but asymptomatic plant material. The use of pathogen-free propagating material as well as planting of resistant genotypes are currently the only strategies used for the disease control. Therefore, knowledge of the phylotype composition and genetic variability in populations of this bacterium is useful for implementing effective control measures. In this work, we developed an efficient intercalating dye-based real-time PCR protocol to detect the bacterium in asymptomatic eucalypt plants as well as to investigate its movement in tissues of clones with different levels of resistance. In addition, we identified phylotypes, sequevars and genotypes of 93 Ralstonia isolates, obtained from cuttings and eucalypt trees grown in different regions of Brazil. Subsequently, sequevar distribution patterns and their relationship with the aggressiveness of the bacterium in eucalypt clones were investigated. Through the real-time PCR technique, we found that the bacterium translocates acropetally and basipetally in inoculated but asymptomatic plants of the resistant clone as in plants of the symptomatic susceptible one. Nevertheless, a lower bacterial concentration was detected in the tissues of the resistant clone, where, through the scanning electron microscope, no bacterial biofilm was observed obstructing the xylem vessels. With the molecular characterization, we found that, like R. solanacearum (phylotype II), R. pseudosolanacearum (phylotype I) also causes eucalypt wilt. However, R. pseudosolanacearum is genetically uniform, which indicates a probable recent introduction of this phylotype in the country. Unlike R. pseudosolanacearum, R. solanacearum is phylogenetically diverse, and there is no correlation between sequevar and geographic origin. Isolate aggressiveness varied between the eucalypt clones tested, reinforcing the importance of conducting molecular and aggressiveness characterizations of the pathogen population, in order to support the selection of resistant material. The qPCR method developed in this study could be valuable for pathogen detection during disease diagnosis as well as pathogen quantification in plant tissue. The present study expands the knowledge of the variability of the Ralstonia solanacearum species complex in Eucalyptus spp. Keywords: Bacterial wilt. Real time PCR. Eucalypt.
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- 2021
38. Detection of QTL associated with rust resistance using IBD-based methodologies in exogamic Eucalyptus spp. populations.
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Rosado, Tatiana Barbosa, Tomaz, Rafael Simões, Junior, Marcio Fernandes Ribeiro, Rosado, Antônio Marcos, da Silva Guimarães, Lúcio Mauro, de Araújo, Elza Fernandes, Alfenas, Acelino Couto, and Cruz, Cosme Damião
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EUCALYPTUS diseases & pests , *PLANT populations , *MICROSATELLITE repeats , *EUCALYPTUS grandis , *CROP yields - Abstract
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. [ABSTRACT FROM AUTHOR]
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- 2010
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39. Erwinia psidii - Eucalyptus spp.: colonização, variabilidade genética, agressividade e teste imunológico para a detecção do patógeno
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Montoya Estrada, Claudia Nohemy, Guimarães, Lúcio Mauro da Silva, Pacheco, Jorge Luis Badel, Oliveira, Leandro Licursi de, and Alfenas, Acelino Couto
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Seca-de-ponteiros ,Bactérias - Detecção ,Morte descendente ,Virulência (Microbiologia) ,Psidium guajava ,Fitopatologia ,Doenças bacterianas das plantas ,Anticorpos policlonais - Abstract
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Seca de ponteiros, causada por Erwinia psidii é atualmente uma das doenças emergentes mais severas na cultura do eucalipto. Apesar de sua importância econômica, em virtude dos riscos que pode causar na eucaliptocultura, pouco se sabe sobre este patossistema. Assim, neste trabalho, a partir de cortes histológicos, estudou-se o processo de colonização em plantas de eucalipto usando um isolado de E. psidii transformado com gfp, estimou-se a variabilidade genética das populações do patógeno por meio de marcadores moleculares rep– PCR (ERIC, REP e BOX) e desenvolveu-se um imunoteste para detecção rápida de E. psidii usando colônias puras isoladas de plantas sintomáticas e assintomáticas. Neste estudo, conseguiu-se transformar E. psidii com pGreen-TIR e demonstrar que o plasmídeo é estável na ausência de seleção com antibióticos in vitro e in vivo. Usando microscopia de fluorescência, demonstrou-se que a colonização de E. psidii nos tecidos não está restrita ao ponto de inoculação (axila foliar). E. psidii coloniza os vasos do xilema, esclerênquima e parênquima das folhas e caule do eucalipto. Aos 35 dias após a inoculação (dai), a bactéria encontrava-se a 5 cm acima do ponto de inoculação, indicando que foi capaz de colonizar a planta acropetalmente, acompanhando o fluxo da água e nutrientes. Análises por microscopia confocal de amostras de plantas inoculadas via sistema raicular revelaram que E. psidii penetra e coloniza raízes primárias e secundárias e atinge os vasos do xilema. No entanto, nos tempos após inoculação avaliados neste estudo, a bactéria ficou restrita nas raízes e não atingiou o caule da planta. Estudos adicionais avaliando tempos após inoculação maiores devem ser realizados para confirmar esses resultados. Acredita-se que E. psidii seja disseminada a partir de mudas infectadas assintomáticas. Análises moleculares (rep-PCR) de 101 isolados de E. psidii obtidos de Eucalyptus spp. e cinco isolados de goiabeira (Psidium guajava) indicam que as populações de E. psidii do Brasil apresentam baixa variabilidade genética. Observou-se, contudo, o agrupamento dos isolados por espécie hospedeira, embora dois isolados de goiaba (LPF681 e LPF682) agruparam-se com os de eucalipto (LPF609). Testes de Wilcoxon e ANOVA dos dados de severidade e AACPD para diferentes isolados da bactéria inoculados em dois clones de eucalipto indicaram que há interação isolado × clone. A AACPD e a severidade da doença variaram significativamente entre isolados e nos dois clones testados. A variabilidade em agressividade entre isolados de E. psidii demonstraram a importância do emprego de isolados mais agressivos para a seleção de genótipos de eucalipto resistentes para plantio em escala comercial. O anti-soro contra o isolado LPF534 de E. psidii (Anti-Ep), obtido neste estudo, reagiu positivamente contra todos os isolados de E. psidii testados, incluindo alguns isolados de goiaba. O teste de aglutinação desenvolvido com este anti-soro é importante para o diagnóstico de E. psidii, por representar uma alternativa rápida e de baixo custo, em comparação com métodos baseados em PCR para a detecção do patógeno em plantas assintomáticas. Dieback, caused by Erwinia psidii is currently one of the most severe emerging diseases in eucalypt plantations. Despite its economic importance, due to the risks posed to eucalypt production, little is known about this pathosystem. In this work, we studied the plant colonization by a gfp-transformed E. psidii isolate using histological sections, estimated the genetic variability of pathogen populations using molecular rep–PCR markers (ERIC, REP and BOX) and developed an immunoassay for the rapid detection of E. psidii in symptomatic and asymptomatic plants. In this study, we were able to transform E. psidii with pGreen-TIR and to demonstrate that the plasmid is stable in the absence of antibiotic selection both in vitro and in vivo. Using fluorescence microscopy, it was possible to demonstrate that tissue colonization by E. psidii is not restricted to the inoculation point (leaf axil) and that it colonizes the xylem vessels, sclerenchyma and parenchyma of the leaves and stem of eucalypt. At 35 days after inoculation (dai), the bacterium was found at 5 cm above the inoculation point, indicating that it was able to colonize the plant acropetally, following the water flow. Confocal microscopy analysis of plant samples inoculated via radicular system revealed that E. psidii penetrates and colonizes primary and secondary roots and reaches the xylem vessels. However, at the times after inoculation evaluated in this study, the bacterium was restricted to the roots and did not reach the stem of the plant. Further studies with longer times after inoculation must be performed to confirm these results. It is believed that E. psidii is disseminated from infected asymptomatic cuttings. Molecular analyzes (rep-PCR) of 101 E. psidii isolates obtained from eucalypt (Eucalyptus spp.) and five guava (Psidium guajava) demonstrated that the populations in Brazil have low genetic variability. Nonetheless, grouping of isolates by host plant was observed, although two guava isolates (LPF681 and LPF682) grouped with eucalypt isolates (LPF609). The Wilcoxon and ANOVA tests on disease severity and AUDPC data indicated an isolate × clone interaction. AUDPC and disease severity varied significantly among isolates and between the two clones tested. The variability in aggressiveness among isolates of E. psidii demonstrated the importance of using the most aggressive isolates in the selection of resistant eucalypt genotypes for commercial scale planting. An antiserum against E. psidii isolate LPF534 (Anti-Ep) obtained in this study reacted positively against all strains of E. psidii tested, including some isolated from guava. The agglutination test designed using this antiserum can be considered important for the diagnosis of E. psidii, as it represents a rapid and low-cost alternative compared to PCR-based methods for detecting the pathogen in asymptomatic plants.
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- 2018
40. Comparative genomic and transcriptomic analyses reveal different pathogenicity-related genes among three eucalyptus fungal pathogens.
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Santos, Samuel A., Vidigal, Pedro M.P., Thrimawithana, Amali, Betancourth, Blanca M.L., Guimarães, Lúcio M.S., Templeton, Matthew D., and Alfenas, Acelino C.
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EUCALYPTUS , *PHYTOPATHOGENIC microorganisms , *GENE families , *TRANSFER RNA , *RIBOSOMAL RNA , *GENES , *COMPARATIVE genomics - Abstract
• Ceratocystis fimbriata LPF1912 has a compact genome with 43% as coding sequencing. • A set of 4919 orthologous gene clusters represent the minimum quorum for the three pathogens. • Two Ceratocystis pathogens showed a different pathogenicity-related genes compared to Calonectria pseudoreteaudii. • The most of DEGs of C. fimbriata LPF1912 is related to its pathogenicity in Eucalyptus spp. Ceratocystis fimbriata is an important plant pathogen known to cause Ceratocystis Wilt (CW), a prevalent fungal disease known to affect Eucalyptus spp. plantations in Brazil. To better understand the molecular mechanisms related to pathogenicity in eucalyptus, we generated a high-quality assembly and annotation of the Ce. fimbriata LPF1912 isolate (LPF1912) genome, as well as the first transcriptome of LPF1912 from 16 eucalyptus clones at three infection incubation periods (12, 18, and 24 h). The LPF1912 genome assembly contains 805 scaffolds, totaling 31.8 Mb, with 43% of the genome estimated to be coding sequence comprised of 7,390 protein-coding genes of which 626 (8.5%) were classified as secreted proteins, 120 ribosomal RNAs, and 532 transfer RNAs. Comparative genomic analysis among three eucalyptus fungal pathogens (Ce. fimbriata, Ce. eucalypticola , and Calonectria pseudoreteaudii), showed high similarity in the proteome (21.81%) and secretome (52.01%) of LPF1912 and Ce. eucalypticola. GO annotation of pathogenicity-related genes of LPF1912 and Ce. eucalypticola, revealed enrichment in cell wall degrading enzymes (CWDEs), and lipid/cutin metabolism for Ca. pseudoreteaudii. Additionally, a transcriptome analysis between resistant and susceptible eucalyptus clones to CW infection indicated that a majority (11) of LPF1912 differentially expressed genes had GO terms associated with enzymatic functions, such as the polygalacturonase gene family, confirming the crucial role of CWDEs for Ce. fimbriata pathogenicity. Finally, our genomic and transcriptomic analysis approach provides a better understanding of the mechanisms involved in Ce. fimbriata pathogenesis, as well as a framework for further studies. [ABSTRACT FROM AUTHOR]
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- 2020
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41. Etiologia e análise de novo do transcriptoma do patógeno causador de oídio em Eucalyptus no Brasil
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Fonseca, Natália Risso, Guimarães, Lúcio Mauro da Silva, and Alfenas, Acelino Couto
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Fitopatologia ,Oídio ,Podosphaera pannosa ,Eucalipto - Doenças e pragas ,RNA ,Eucalyptus urophylla - Abstract
Conselho Nacional de Desenvolvimento Científico e Tecnológico Oídio do eucalipto é uma importante doença que ocorre principalmente em casas de vegetação e minijardins clonais protegidos de eucalipto (Eucalyptus spp.) no Brasil. O fungo infecta folhas jovens e brotações. Sobre o tecido afetado, observam-se colônias superficiais isoladas ou agrupadas do fungo de coloração branca, que podem atingir toda superfície foliar e induzir malformação dos órgãos afetados e resultar em redução do crescimento e da produção de brotos para estaquia. Devido ao aumento da incidência e importância dessa doença nos últimos anos e também à falta de pesquisas relacionadas a esse patossistema, esse estudo objetivou: i) determinar a etiologia do oídio do eucalipto por meio do sequenciamento da região ITS e 28S do rDNA e de análises morfológicas de isolados de oídio coletados em diferentes regiões geográficas do Brasil; e ii) analisar o transcriptoma do patógeno durante a infecção em Eucalyptus urophylla gerado pelo sequenciamento do transcriptoma (RNA-Seq) e montagem de novo. Baseado nos resultados de análises filogenéticas e caracterização morfológica, todos os 42 isolados coletados foram identificados como Podosphaera pannosa, também conhecido como agente etiológico do oídio das roseiras. Inoculações cruzadas com isolados de P. pannosa de roseira e eucalipto demonstraram que P. pannosa pode infectar ambas as espécies. O sequenciamento do transcriptoma de P. pannosa pela plataforma Illumina resultou em 12.107 transcritos. Entre os 10 transcritos mais abundantes, encontram-se os genes codificadores de enzimas envolvidas no estabelecimento e crescimento do fungo. A predição do secretoma do fungo resultou em 217 proteínas, das quais 14 foram consideradas como candidatas a efetores. Além disso, 242 pares de primers foram desenhados a partir das sequências do transcriptoma com potencial para amplificar regiões microssatélites (Simple Sequence Repeats - SSR) de P. pannosa. Os resultados gerados neste trabalho demonstram que apenas a espécie P. pannosa causa doença no eucalipto. Além disso, fornece informações úteis para novos avanços nos estudos sobre a doença por oferecer uma base para a melhor compreensão sobre o patossistema P. pannosa- eucalipto. Eucalypt powdery mildew is an important disease that occurs mainly in greenhouses and protected clonal hedges of eucalypt (Eucalyptus spp.) in Brazil. The fungal pathogen infects new leaves and shoots. White superficial colonies isolated or grouped that grow over the affected plant tissue are observed, which can subsequently spread to all leaf surface, causing leaf malformation and reduction on growth and production of shoots for mini-cutting. Because this disease has increased in incidence and importance in recent years, and research on this pathosystem is largely lacking, the objectives of this study were to i) determine the etiology of the disease through ITS and 28S rDNA sequencing and morphological analyses of powdery mildew pathogens isolates collected in different regions in Brazil; and ii) analyze the transcriptome of the powdery mildew pathogen during infection on Eucalyptus urophylla generated by RNA sequencing (RNA-Seq) and de novo assembly. Based on the results of phylogenetic analyses and morphological characteristics, all 42 pathogen isolates collected were identified as Podosphaera pannosa, also known to cause rose powdery mildew. Cross inoculations with pathogen isolates from rose (Rosa spp.) and eucalypt demonstrated that P. pannosa can infect both host species. The transcriptome sequencing by Illumina platform resulted in 12,107 transcripts. Among the 10 most abundant transcripts included genes encoding enzymes involved in fungal establishment and growth. The secretome prediction resulted in 217 proteins, of which 14 were considered as candidate effectors. In addition, 242 primer pairs were designed from the transcriptome sequences with potential to amplify P. pannosa microsatellites (Simple Sequence Repeats – SSR) regions. The results demonstrate that P. pannosa is the only causal agent found for eucalypt powdery mildew. In addition, this study provides technological advances in the study of this disease that will provide a basis for a better understanding of the P. pannosa- eucalypt pathosystem.
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- 2016
42. Análise de ligação e mapeamento de QTLs em populações simuladas
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Alves, Alexandre Alonso, Cruz, Cosme Damião, Resende, Marcos Deon Vilela de, Alfenas, Acelino Couto, Bhering, Leonardo Lopes, and Guimarães, Lúcio Mauro da Silva
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QTL mapping ,Statistical genomics ,Mapeamento genético ,Mapeamento de QTLs ,Genetic mapping ,CIENCIAS BIOLOGICAS::GENETICA::GENETICA QUANTITATIVA [CNPQ] ,Estatística genômica - Abstract
Conselho Nacional de Desenvolvimento Científico e Tecnológico Como os recentes avanços na tecnologia têm levado ao desenvolvimento de novas tecnologias de genotipagem, no futuro, é mais provável que os mapas de ligação de alta densidade serão construídos a partir de marcadores dominantes e co-dominantes. Recentemente, uma abordagem estritamente genética foi proposta para a estimação da freqüência de recombinação (r) entre marcadores co-dominantes em famílias de irmãos completos. O conjunto completo de estimadores quase foi obtido, mas infelizmente, uma configuração envolvendo a estimativa da distância entre os marcadores dominantes, que segregam na proporção 3:1 e marcadores co-dominantes, não foi levada em consideração. Aqui novos nove estimadores são acrescentados ao conjunto previamente publicado, tornando possível cobrir todas as combinações de marcadores moleculares com dois a quatro alelos (sem epistasia) em uma família de irmãos completos. Isso inclui a segregação em um ou ambos os genitores, dominância e todas as configurações de fases de ligação. Como populações de retrocruzamentos (RC) são frequentemente utilizadas como populações de mapeamento, tanto em espécies autógamas, quanto em espécies alógamas foi conduzido um estudo de simulação para testar as implicações do tamanho da população, herdabilidade da característica, propriedades do QTL (r2, a e posição) e densidade de marcadores no poder de detecção e precisão do mapeamento de QTLs. Para tanto foram simuladas populações com diferentes tamanhos, com diferentes características (h2, número de QTLs e posição) e os dados analisados com dois métodos de mapeamento de QTLs comumente utilizados (mapeamento por intervalo simples (MIS) e mapeamento por intervalo composto (MIC)). Verificou-se que o tamanho da amostra tem uma grande implicação no poder de detecção e como conseqüência na estimação da magnitude da variação explicada pelo QTL e no efeito genético, em função de populações pequenas não permitirem o mapeamento de QTLs de pequeno efeito, principalmente quando esses estão envolvidos no controle genético de características de baixa herdabilidade. Também foi verificado que o posicionamento de QTLs baseados em MIC é mais acurado que MIS e que em média os QTLs mapeados estavam próximos as suas posições simuladas. Um resultado interessante é que o MIC tende a subestimar os valores de magnitude (r2) especialmente em populações grandes/ características de baixa herdabilidade e superestimá-la em populações pequenas, o que pode ser um reflexo do pequeno coeficiente de variação do erro utilizado, ou devido ao fato de quando os marcadores não se encontram na exata posição do QTL, esse parâmetro é de fato esperado ser subestimado. Destaca-se também, o fato que quando marcadores estão amplamente distribuídos ao longo do genoma (~10cM), e desse modo cobrindo a região do QTL, se um dos marcadores já estiver próximo ao QTL, um maior número de marcadores (~1cM) não melhora a precisão do mapeamento do QTL em populações suficientemente grandes. Baseado nesses resultados recomenda-se o uso de populações de tamanho adequado, ≥500, se a intenção é mapear QTLs em populações de RC, porque nessa situação, mesmo mapas de média densidade podem ser usados para mapear QTLs de grande ou pequeno efeito com grande confiabilidade. Finalmente, como os procedimentos de mapeamento de ligação e mapeamento de QTLs em famílias de irmãos completos (FIC) de espécies alógamas são bastante diversos, foi conduzido um estudo comparando o método de mapeamento por pseudo-testcross modificado (PST) (usando microsatélites), com o método de mapeamento baseado na FIC; em termos de ordenamento dos marcadores, distância entre os marcadores, comprimento total do mapa, variância das estimativas de distância e estresse. Investigou-se também o poder de detecção e a precisão de métodos de mapeamento de QTLs por intervalos baseados nos mapas PST ou no mapa para a FIC. Verificou-se que em geral as duas estratégias geram mapas altamente correlacionados com comprimentos dos grupos de ligação proporcionais. Verificou-se também que independentemente da abordagem de mapeamento de QTLs utilizadas, o poder de detecção é reduzido em populações pequenas, especialmente em situações onde a herdabilidade da característica ou magnitude do QTL é pequena. Também foi verificado que apesar dos dois métodos serem aparentemente equivalentes em termos de posicionamento do QTL para características de alta herdabilidade/ QTLs de grande efeito, o MIC baseado nos mapas pseudo-testcross prove dados mais acurados para características de baixa herdabilidade/QTLs de pequeno efeito. Como relação à magnitude dos QTLs, notou-se que ambos os métodos parecem ser equivalentes, sendo os valores superestimados para características de alta herdabilidade e subestimados para características de baixa herdabilidade, independentemente do tamanho amostral. Assim para espécies alógamas com médio nível de recursos genômicos, propõem-se que tanto a abordagem de PST quanto a abordagem baseada na FIC, e métodos de mapeamento de QTLs relacionados, possam ser utilizados para gerar mapas genéticos e mapear QTLs com alta confiabilidade. É importante ressaltar, entretanto, que outros estudos, usando diferentes cenários, i.e. diferentes coeficientes de variação do erro, diferentes números de QTLs, diferentes distribuições de marcadores, que coletivamente podem tornar a simulação um pouco mais realística, são necessários para verificar que os resultados deste trabalho se mantêm em todas as situações. As high-throughput genomic tools have led to the development of novel genotyping procedures, it is likely that, in the future, high density linkage maps will be constructed from both dominant and co-dominant markers. Recently, a strictly genetic approach was described for estimating the recombination frequency (r) between co-dominant markers in full-sib families. The complete set of maximum likelihood estimators for r in full-sib families was almost obtained, but unfortunately, one particular configuration involving dominant markers, segregating in a 3:1 ratio and co-dominant markers, was not considered. Here we add nine further estimators to the previously published set, thereby making it possible to cover all combinations of molecular markers with two to four alleles (without epistasis) in a full-sib family. This includes segregation in one or both parents, dominance and all linkage phase configurations. As backcross (BC) populations are often used as mapping populations both in self pollinating species, and in out-breeding species we also undertook a simulation study to test implications of population size, trait heritability, QTL properties (r2, a and position) and marker density in the power and precision of QTL mapping. For that we have simulated populations with different sizes, with different characteristics (h2, QTL number and location) and the data analyzed with two xv QTL mapping methods (simple interval mapping (SIM) and composite interval mapping (CIM)). We found that sample size has a major implication in the detection power and as consequence in the estimation of the magnitude and additive genetic effect, as small populations do not allow mapping of low effect QTLs, especially if these QTLs are involved in the genetic control of traits with low heritability. We also found that the positioning of the QTLs based on CIM is more accurate than SIM and that on average the mapped QTLs are close to their simulated position. The results showed that CIM tend to underestimate the magnitude (r2) values especially in large population sizes/low heritabilities traits and overestimate it in smaller populations, which can be a reflection of the low coefficient of variation of the error used, or due to fact that when markers aren´t in the same of the QTL, this parameter is indeed expected to be underestimated. We also highlight the fact, that when markers are evenly distributed across the genome (~10 cM), and therefore covering the QTL region, if one of the markers is already close to the QTL, larger number of markers (~ 1cM) do not improve the precision of QTL mapping in sufficiently large mapping populations. Based on our results we recommend the use of adequate sample size, say ≥500, if the intention is map QTLs in backcross populations, because in this situation even mid-density genetic maps can be used to map QTLs of large or small effect with high confidence. Finally, as the procedures for linkage and QTL mapping in fullsib families of outbreeding species are quite diverse, we also undertook a simulation study comparing the modified pseudo-testcross (using SSR markers) and the full-sib mapping designs in terms of marker ordering, distance between markers, total map size, distance variance and stress. We also investigated the power and precision of interval mapping procedures based on the full-sib and on the modified pseudo-testcross maps. We show that in general the modified pseudo-testcross and the full-sib mapping designs generate highly correlated maps with proportional linkage groups length. That independent of the QTL mapping approach used, detection power is reduced in small populations, especially in situations where trait heritability or QTL magnitude are low. We also found that although both methods appear to be equivalent in terms of QTL positioning for high heritability traits/major effect QTLs, the CIM based on modified pseudo-testcross maps provide more accurate data for low heritability traits/minor effect QTLs in larger populations. With regard to QTLs magnitude, we show that both methods appear to be equivalent, and that the magnitude values tended to be over estimated for the high heritability trait, and underestimated for the low heritability trait, independent of the sample size. Thus, for outbreeding species with mid-level of genomic resources we propose that either the modified pseudo-testcross or the single full-sib mapping design and the related QTL mapping strategies can be used to generate genetic maps and map QTLs with high confidence. It is important to highlight however, that, other studies, using different scenarios, i.e. different coefficients of variation of the error, different number of QTLs, different marker distributions, which collectively may make the simulation a bit more realistic, are needed in order to see if the results of our work hold true in every situation.
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- 2010
43. Estrutura genética de populações de Ceratocystis fimbriata e padrão espaço-temporal da murcha-de-ceratocystis
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Ferreira, Maria Alves, Maffia, Luiz Antônio, Jesus Júnior, Waldir Cintra de, Alfenas, Acelino Couto, Harrington, Thomas Charles, Guimarães, Lúcio Mauro da Silva, and Zauza, Edival ângelo Valverde
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Estrutura genética ,CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA [CNPQ] ,Ceratocystis fimbriata ,Genetic structure ,Ceratocystis wilt ,Murcha-de-ceratocystis - Abstract
Conselho Nacional de Desenvolvimento Científico e Tecnológico Ceratocystis fimbriata é um patógeno que causa murcha vascular e tem ocasionado perdas em muitas culturas importantes no Brasil, incluindo Eucalyptus spp., Mangifera indica, Ficus carica, Colocasia esculenta e Gmelina arborea. Suspeita-se que algumas populações de C. fimbriata no Brasil sejam espécies crípticas, hospedeiro-especializadas. Foram usados 15 marcadores polimórficos para comparar a diversidade genética entre 13 populações de C. fimbriata coletadas em diferentes espécies hospedeiras, em seis Estados brasileiros. Adicionalmente, foram examinadas 20 populações de C. fimbriata coletadas nos plantios clonais de eucalipto nos Estados de São Paulo (SP), Bahia (BA) e Minas Gerais (MG), usando seis marcadores polimórficos. Também foi avaliado o padrão espacial da doença em 35 parcelas em diferentes plantios clonais de eucalipto, entre estas, quatro áreas de coleta de brotações para enraizamento. Os valores de diversidade gênica da maioria das populações em eucalipo e mangueira dos Estados de MG, BA, Rio de Janeiro (RJ) e SP foram similares aos de populações possivelmente naturais de C. platani e C. cacaofunesta, espécies pertencentes ao complexo C. fimbriata. Valores de índice de associação evidenciaram desequilíbrio de ligações entre as populações do fungo originárias de eucalipto e mangueira. As populações e os genótipos do fungo isolados de eucalipto e mangueira aparentemente estão ligados entre si de acordo com as análises de UPGMA. Em contraste, a população de G. arborea do Pará e F. carica de SP tiveram baixos níveis de diversidade e foram diferentes entre si, e de todas as populações estudadas, sugerindo que elas têm origens diferentes. Algumas populações apresentaram um único genótipo e podem ter sido introduzidas na região por mudas infectadas. Constatou-se, com base nas análises genéticas e de fertilidade, que as populações brasileiras de C. fimbriata pertencem a uma única espécie biológica. Dos 177 isolados analisados em 20 populações do fungo isoladas de plantios clonais de eucalipto em SP, BA e MG, foram identificados 79 genótipos. As diversidades genéticas nas populações de MG e em lgumas da BA foram similares ao esperado para populações nativas do patógeno. Entretanto, algumas das populações apresentaram baixa ou ausência de diversidade genética. Alta diversidade genética foi encontrada nas áreas em que havia pequena propriedade ou floresta do tipo Cerrado antes da cultura do eucalipto, sugerindo que C. fimbriata estava estabelecido no solo antes do plantio dessa cultura. Assim, strains de C. fimbriata isolados de eucalipto parecem ser nativos de algumas áreas de MG e BA. Em contraste, um ou somente poucos genótipos foram encontrados em áreas em que havia pastagens anteriormente ao eucalipto. Adicionalmente, esses genótipos foram encontrados em viveiros ou áreas de coleta de brotações. As populações de C. fimbriata de SP foram relacionadas a algumas das populações da BA, e muitos genótipos encontrados em SP também foram observados na BA. Desse modo, o patógeno pode ter sido introduzido em novas áreas por meio de mudas contaminadas. O padrão espaçotemporal da murcha-de-ceratocysits foi avaliado em 35 parcelas em plantios de eucalipto. A maioria dos plantios estudados apresentou padrão agregado da doença. Assim, pode-se inferir que os plantios podem ter sido infectados via inóculo do solo. Entretanto, em alguns casos, a distribuição de mudas contaminadas pode ter influenciado o padrão espacial da doença no campo. Ceratocystis fimbriata is an insect-associated wilt pathogen that causes serious losses on many important crops in Brazil, including Eucalyptus spp. (eucalyptus), Mangifera indica (mango), Ficus carica (fig), Colocasia esculenta (inhame), and Gmelina arborea. It was suspected that some populations of C. fimbriata in Brazil are host-specialized, cryptic species, but mating studies showed that isolates of C. fimbriata from these hosts and sweet potato are interfertile, and progeny from those crosses showed normal segregation for microsatellite markers. We used 15 highly-polymorphic microsatellite markers to compare genetic diversity among 13 populations of C. fimbriata collected from six states in Brazil. The gene diversity values of most eucalyptus and mango populations from Minas Gerais, Bahia, Rio de Janeiro, and São Paulo were similar to putatively-natural populations of C. platani and C. cacaofunesta, two other species in the C. fimbriata complex that are homothallic. Index of association values indicated substantial asexual reproduction or selfing in populations on mango and eucalyptus. Most of these eucalyptus and mango populations were not highly differentiated from each other, there was evidence of gene flow among them, and the populations and genotypes appeared to be related to each other by UPGMA analyses. In contrast, the Gmelina population from Pará and the fig and inhame populations from São Paulo had relatively low levels of diversity and were highly differentiated from each other and all other studied populations, suggesting that they were from different origins and had gone through genetic bottlenecks. Twenty populations of C. fimbriata from clonal plantations of eucalyptus in São Paulo, Bahia and Minas Gerais were examined using six polymorphic microsatellite markers. We identified 79 genotypes among the 177 isolates of C. fimbriata from individual trees in the 20 plantations. The gene and genotypic diversity values of eucalyptus populations (plantations) from Minas Gerais and some populations in Bahia were similar to those expected for natural, soilborne populations. However, some of the other populations showed little or no genetic diversity. Populations of high genetic diversity data were found in plantations that were formerly farmland or forest, suggesting that C. fimbriata was established in the soil before eucalyptus was planted. In contrast, one or only a few genotypes were found in some plantations on previous pastureland (with no woody hosts for C. fimbriata), and these same genotypes were found in nurseries or plantations that were the source for rooted cuttings. São Paulo populations of C. fimbriata were closely related to some of the Bahia populations, and many of the genotypes found in São Paulo were also found in Bahia, often on the same eucalyptus clones. Thus, although eucalyptus strains of C. fimbriata appear to be native to some areas of Minas Gerais and Bahia, the pathogen apparently can be introduced to new areas in infected eucalyptus cuttings from diseased mother plants taken from plantations or hedges in nurseries. The spatial-temporal patterns of Ceratocystis wilt were evaluated in 35 plots of eucalyptus plantations. In the majority of plantations studied, Ceratocystis wilt occurred in clusters. In many of the plantations, it appeared that the eucalyptus trees were infected from soilborne inoculum. However, the distribution of contaminated rooted in the field can influence the spatial partern of the disease.
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- 2009
44. First Draft Genome Sequence of Xanthomonas axonopodis pv. eucalyptorum, Causal Agent of Bacterial Leaf Blight on Eucalypt.
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Neves YF, Santos SA, Guimarães LMS, Vidigal PMP, Badel JL, Alfenas-Zerbini P, Mafia RG, and Alfenas AC
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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., (Copyright © 2019 Neves et al.)
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- 2019
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45. Draft Genome Sequence of Erwinia psidii, Causal Agent of Bacterial Blight of Guava ( Psidium guava ) and Dieback of Eucalypt ( Eucalyptus spp.).
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Hermenegildo PDS, Santos SA, Guimarães LMS, Pereira IC, Vidigal PMP, Badel JL, Alfenas-Zerbini P, Mafia RG, Ferreira MASV, and Alfenas AC
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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.
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- 2019
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46. Xanthomonas axonopodis pv. eucalyptorum pv. nov. Causing Bacterial Leaf Blight on Eucalypt in Brazil.
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Ferraz HGM, Badel JL, da Silva Guimarães LM, Reis BP, Tótola MR, Gonçalves RC, and Alfenas AC
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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.
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- 2018
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47. Rhizobacterial characterization for quality control of eucalyptus biogrowth promoter products.
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Zarpelon TG, Guimarães LM, Alfenas-Zerbini P, Lopes ES, Mafia RG, and Alfenas AC
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- Anti-Bacterial Agents pharmacology, Microbial Sensitivity Tests, Polymorphism, Restriction Fragment Length, RNA, Ribosomal, 16S genetics, Bacteria classification, Bacteria drug effects, Bacteria genetics, Bacteria growth & development, Eucalyptus growth & development, Eucalyptus microbiology, Rhizosphere
- 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 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., (Copyright © 2016. Published by Elsevier Editora Ltda.)
- Published
- 2016
- Full Text
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48. Genetic control of Eucalyptus urophylla and E. grandis resistance to canker caused by Chrysoporthe cubensis.
- Author
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da Silva Guimarães LM, de Resende MD, Lau D, Rosse LN, Alves AA, and Alfenas AC
- Abstract
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.
- Published
- 2010
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