In Arabica coffee breeding, some of the most used sources of resistance to leaf rust (Hemileia vastatrix) are natural Coffea arabica x canephora hybrids ("Híbrido de Timor"). To decipher the cellular and molecular nature of that resistance, leaves of genotype HDT832/2, were challenged with H. vastatrix race II, and monitored using light microscopy and RT-qPCR expression analysis of genes involved in plant immunity mechanisms (receptor-like kinase, WRKY transcription factor 1, phenylalanine ammonia-lyase, chalcone synthase, 13-lipoxygenase, glycosyltransferase, pathogenesis related PR1b and PR10). These were compared to the nonhost resistance responses of HDT832/2 to the infection by the cowpea rust fungus (Uromyces vignae). H. vastatrix ceased growth more frequently after stomata penetration, forming few haustoria, inducing a hypersensitive-like response, phenol accumulation and haustorium encasement with callose. U. vignae could enter stomata but failed to form haustoria, while inducing hypersensitive-like responses and phenol accumulation. In host and nonhost interactions, activation of genes involved in signalling coincided with the differentiation of appressoria, and cellular responses (hypersensitive-like responses and accumulation of phenolic compounds) were recorded from the full appressorium or penetration hypha stages onwards. Similarly, a gene related to the JA pathway was first activated at the penetration hypha stage for both interactions, while genes related to the SA pathway were only activated in the host interaction, the latter being the single clear difference between host and nonhost interactions. The cellular and molecular resistance responses of HDT832/ 2 to these rust fungi suggest that common immunity components are shared between host and nonhost resistance, which may explain the longer durability of this resistance.
The virulence of two wild type (PA45 and PA37) and two genetically modified (13C: hygromycin resistant; FATSS: hygromycin resistant and β-cin knock-down) Phytophthora cinnamomi strains towards cork oak (Quercus suber) was assessed via a quantitative evaluation of disease symptoms arising from a soil infestation assay, and by a histological analysis of root colonization. Comparison of virulence, as expressed by symptom severity, resulted in the following ranking: highly virulent (wild type strains), medium virulence (strain 13C) and weakly virulent (FATSS). Both transgenic strains were compromised in their virulence, as expressed by symptom severity, but strain 13C was much less affected than FATSS. Microscopic observation showed that the FATSS strain was unable to effectively invade the root, while 13C and the two wild type strains were all able to rapidly colonize the whole root, including the vascular tissue. These results strengthen the notion that elicitins are associated, either directly or indirectly, with the infection process of Phytophthora.
Leaf rust, caused by Hemileia vastatrix Berk & Broome, is the most destructive fungal disease of coffee. In the absence of a suitable gene validation system in coffee, the objective of this study was to investigate whether the model plant Arabidopsis thaliana (L.) Heynh. may be used as a heterologous system for the molecular dissection of coffee responses to leaf rust. Histological examination of A. thaliana (Col-0) leaves inoculated with H. vastatrix (race II) showed that by 24 h after inoculation (hai), H. vastatrix uredospores differentiated appressoria and penetrated the stomata, but failed to form haustoria. Arabidopsis thaliana cellular resistance responses included hypersensitive-like response (HR) of stomata guard cells together with accumulation of phenolic compounds and callose deposition in walls of epidermal and mesophyll cells. Results indicate that H. vastatrix infection triggered the induction of a set of defence-related genes peaking at 18 and 42 hai. The non-host HR triggered by H. vastatrix in the model plant A. thaliana makes it usable to infer the function of coffee genes involved in pre-haustorial rust resistance.
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