Summary A new disease of unknown bacterial aetiology has been observed in eucalyptus stands since 2009. It is characterized by die‐back, wilting and lesions on the branches, petiole and midrib in association with macroscopic and microscopic bacterial ooze. To date, this disease has been observed in stands of clonal Eucalyptus saligna, E. grandis and E. urophylla x E. grandis hybrids and in E. dunnii seedling plantations in the states of São Paulo, Rio Grande do Sul and Mato Grosso do Sul. Considering the economic importance of eucalyptus plantations and the potential losses caused by this disease, this study aimed to identify and characterize the causal agent. Thirty‐four strains were obtained from infected plants, which were collected in the field from four locations. The inoculation of detached leaves and intact rooted cuttings supported pathogenicity in eucalyptus. The phylogenetic analysis of four housekeeping genes (16S rDNA, gapA, recA and rpoB) as well as biochemical tests confirmed the identity of strains belonging to the species Erwinia psidii. This is the first report of E. psidii as the cause of wilt and die‐back in Eucalyptus spp. in Brazil.
Among the bacterial pathogens of Eucalyptus in Brazil, Ralstonia solanacearum is considered one of the most important because of the characteristics of the pathogen, like the high diversity among the strains related to host range, high virulence, broad geographical distribution and its damage to the crop in recent years. Given its importance and the lack of research on this pathosystem, the present study aimed to perform a molecular characterization of different strains of infected Eucalyptus plants in Brazil. A total of 19 bacterial cultures isolated from Eucalyptus in different regions of Brazil were analysed. A 372-bp product generated by multiplex-PCR amplification using Nmult primers identified all the strains analysed as belonging to phylotype II. Eighteen strains were grouped into subclade IIA and one into subclade IIB. The phylogenetic tree generated from the gene sequences of endoglucanase (egl) confirmed the classification of the strains into phylotype II and separated the strains into sequevars. Strains AMC22, IBSBF2568 and IBSBF2576 were grouped into a single clade, as were strains UFV18 and UFV20, with 89% and 78% a posteriori probability, respectively, forming two new potential sequevars not yet defined. We also identified strains belonging to sequevars 41 (100% probability) and 37 (88% probability). However, most of the strains did not fit into any previously described sequevar and did not form distinct clades. The results of the analysis of fragments amplified using the ERIC-PCR technique indicated the existence of genetic diversity among the strains studied, with a generally high correlation between similarity and the geographical origin of the strains.
The genetic variability and aggressiveness of Brazilian Erwinia psidii isolates from Eucalyptus spp. was studied and compared with reference isolates from guava (Psidium guajava). Repetitive element sequence (rep)‐based PCR markers of 101 isolates from Eucalyptus spp. and five from guava showed that the populations of E. psidii displayed a relatively low genetic variability. No correlation of genetic clustering based on rep‐PCR analysis with geographic origin or host of origin was observed, indicating that genome rearrangements associated with adaptation to a particular host were not detected by these molecular markers. A higher genotypic richness was detected in the Mato Grosso do Sul population, probably reflecting a pathogen dissemination associated with the recent expansion in eucalypt plantations. Wilcoxon and ANOVA tests of disease severity data indicated differences in aggressiveness among isolates and an isolate × clone interaction. The area under the disease progress curve (AUDPC) and disease severity for some isolates were significantly different between two susceptible clones tested. Notably, isolate LPF681 from guava was not able to cause disease on a susceptible Eucalyptus urophylla clone, suggesting that some co‐evolution between pathogen and host has taken place. The variability in aggressiveness and virulence among isolates of E. psidii observed in this study will be important for the establishment of appropriate screening approaches to select for disease resistance.
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.
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.
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