The GH61 represents the most enigmatic Glycoside Hydrolase family (GH) regarding enzymatic activity and importance in cellulose degradation. Heterobasidion irregulare is a necrotizing pathogen and white-rot fungus that causes enormous damages in conifer forests. The genome of H. irregulare allowed identification of ten HiGH61 genes. qRT-PCR analysis separate the HiGH61 members into two groups; one that show up regulation on lignocellulosic substrates ( HiGH61A , HiGH61B , HiGH61D , HiGH61G , HiGH61H , and HiGH61I ) and a second showing either down-regulation or constitutive expression ( HiGH61C , HiGH61E , HiGH61F , and HiGH61J ). HiGH61H showed up to 17,000-fold increase on spruce heartwood suggesting a pivotal role in cellulose decomposition during saprotrophic growth. Sequence analysis of these genes reveals that all GH61 s except HiGH61G possess the conserved metal-binding motif essential for activity. The sequences also divide into groups having either an insert near the N terminus or an insert near the second catalytic histidine, which may represent extensions of the substrate-binding surface. Three of the HiGH61 s encode cellulose-binding modules (CBM1). Interestingly, HiGH61H and HiGH61I having CBM1s are up-regulated on pure cellulose. There was a common substrate-specific induction patterns of the HiGH61 s with several reference cellulolytic and hemicellulolytic GHs, this taken together with their low transcript levels on media lacking lignocellulose, reflect the concerted nature of cell wall polymer degradation. Electronic supplementary material The online version of this article (doi:10.1007/s00253-012-4206-x) contains supplementary material, which is available to authorized users.
Fusarium spp. belong to the division Ascomycota and cause important plant diseases; these fungi may contaminate food products with mycotoxins, endangering human and animal health. Several Fusarium spp. have been associated with potato dry rot. The most frequent and devastating of these species are F. sambucinum, F. solani and F. oxysporum, depending on the geographic location and the season. Samples of potato tubers with dry rot symptoms were collected, and their putative fungal isolates were identified as Fusarium species using partial nucleotide sequences of the internal transcribed spacer, translation elongation factor 1-α and β-tubulin genes. Among 149 isolates, 12 species were identified. F. oxysporum was the most frequent (45 % of the isolates), followed by F. avenaceum (12.1 %), F. solani (10.7 %) and F. sambucinum (7.4 %). Phylogenetic analyses confirmed the species identifications and revealed a high diversity of F. solani and a low diversity of F. oxysporum. Potential producers of zearalenone and trichothecenes were identified within the obtained isolates using PCR markers. Isolates that were pathogenic to potatoes in laboratory tests were found in four species: F. sambucinum, F. avenaceum, F. culmorum, and F. graminearum. The effects of increased temperature and mixed inoculum on the pathogenicities of chosen species were evaluated. This study adds 434 potato-derived Fusarium sequences to the NCBI GenBank database and demonstrates that the list of Fusarium species and mycotoxins present in potato tubers may be richer than previously believed, regardless of whether these species cause dry rot or live as saprophytes.
This study describes late blight resistance of potato breeding lines resulting from crosses between cultivar 'Sárpo Mira' and Rpi-phu1 gene donors. The progeny is investigated for the presence of Rpi-Smira1 and Rpi-phu1 resistance (R) genes. Interestingly, in detached-leaflet tests, plants with both R genes withstood the infection of the Phytophthora infestans isolate virulent to each gene separately, due to either interaction of these genes or the presence of additional resistance loci. The interaction was studied further in three chosen breeding lines on the transcriptional level. The Rpi-phu1 expression, measured over 5 days, revealed different patterns depending on the outcome of the interaction with P. infestans: it increased in infected plants whereas it remained low and stable when infection was unsuccessful. The expression patterns of P. infestans effectors Avr-vnt1, AvrSmira1, and Avr8, recognized by the Rpi-phu1, Rpi-Smira1, and Rpi-Smira2 genes, respectively, were evaluated in the same experimental setup. This is the first report that the Avr-vnt1 effector expression is not switched off permanently in virulent isolates to avoid recognition by an R protein but can reappear in a postbiotrophic phase and is present constantly when infecting plants without the corresponding R gene. Both a plant and a pathogen can react to the other interacting side by changing the transcript accumulation of R genes or effectors.
Fusarium is one of the most important genera of phytopathogenic fungi, causing potato wilt in the field and potato tuber dry rot during storage. The objectives of this study were to identify Fusarium species associated with both potato diseases in different growing regions in Algeria, and to assess their pathogenicity. Among the 152 isolates collected from symptomatic potato plants and tubers in different provinces in Algeria, 13 species of Fusarium and Neocosmospora were identified. Among these three species were isolated only from plants showing symptoms of Fusarium potato wilt (F. oxysporum, F. venenatum, Neocosmospora solani). Two species (F. culmorum, N. tonkinensis) and an isolate of Neocosmospora sp. were found exclusively in tubers with potato dry rot and the remaining ones (F. redolens, F. cf. tricinctum, F. sambucinum, F. cf. incarnatum-equiseti, F. nygamai, F. brachygibbosum and N. falciformis) were associated with both sample types. Fusarium sambucinum was the most frequent species (52.6% of isolates). Fusarium oxysporum and F. nygamai isolates were the most aggressive in the potato wilt pathogenicity test, and F. sambucinum isolates were the most aggressive in the potato tuber pathogenicity test. This is the first study identifying and characterizing potato dry rot and potato wilt pathogens in Algeria.
Late blight of potato, caused by Phytophthorainfestans, is one of the most economically important diseases worldwide, resulting in substantial yield losses when not adequately controlled by fungicides. Late blight was a contributory factor in The Great Irish Famine, and breeding for resistance to the disease began soon after. Several disease-resistant cultivars have subsequently been obtained, and amongst them Sárpo Mira is currently one of the most effective. The aim of this work was to extend the knowledge about the genetic basis of the late blight resistance in Sárpo Mira and to identify molecular markers linked to the resistance locus which would be useful for marker-assisted selection. A tetraploid mapping population from a Sárpo Mira × Maris Piper cross was phenotyped for foliar late blight resistance using detached leaflet tests. A locus with strong effect on late blight resistance was mapped at the end of chromosome XI in the vicinity of the R3 locus. Sárpo Mira’s genetic map of chromosome XI contained 11 markers. Marker 45/XI exhibited the strongest linkage to the resistance locus and accounted for between 55.8 and 67.9 % of variance in the mean resistance scores noted in the detached leaflet assays. This marker was used in molecular marker-facilitated gene pyramiding. Ten breeding lines containing a late blight resistance locus from cultivar Sárpo Mira and the Rpi-phu1 gene originating from the late blight resistant accession of Solanumphureja were obtained. These lines have extended the spectrum of late blight resistance compared with Sárpo Mira and it is expected that resistance in plants containing this gene pyramid will have enhanced durability.
The goal of this study was to determine the expression profile of a late blight resistance gene, Rpi-phu1, after challenge with an incompatible Phytophthora infestans isolate. Relative expression levels were measured by quantitative real-time PCR, before as well as 1, 3 and 5 days after contact with the pathogen. Plants tested included diploid and tetraploid breeding lines with the gene in different genetic backgrounds and plants of a chosen breeding line at different developmental stages. All tested potato breeding lines with the Rpi-phu1 gene were highly resistant to P. infestans and so was the line tested at different developmental stages. No significant changes were detected in Rpi-phu1 transcript level after pathogen challenge in any of the tested tetraploid potato lines. However, Rpi-phu1 transcription in diploid lines was enhanced by contact with P. infestans. Basal expression of the Rpi-phu1 gene was significantly lower in the youngest, 3-week-old plants than in 6-week-old plants. Nevertheless, 1 day after inoculation with the pathogen, transcript levels in these plants were slightly greater than in 6-week-old plants, indicating that plant age can influence transcription of the Rpi-phu1 gene, but that this does not affect late blight resistance.
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