The white collar complex is essential for sexual reproduction but dispensable for conidiation and invasive growth in Fusarium verticillioides

Fvwc1 and Fvwc2, orthologues of the wc-1 and wc-2 genes encoding for proteins of the white collar complex (WCC) in Neurospora crassa were cloned from Fusarium verticillioides and lack-of-function wc mutants were obtained by targeted gene disruption. Photo-conidiation was found to be absent in F. verticillioides, on the contrary, the wild type strain produced less conidia under continuous illumination than in the dark. Inactivation of any of the wc genes led to total female sterility, without affecting male fertility or asexual conidiation. No loss in colonization capability/invasive growth of the wc mutants was observed, when assessed on tomato fruits. Both Fvwc1 and Fvwc2 showed constitutive expression in the wild type cultures incubated in the dark and exposure to light caused only negligible increases in their transcription. Both Fvwc1 and Fvwc2 were down-regulated in a ΔFvmat1-2-1 gene disruption mutant, lacking a functional mating type (mat1-2-1) gene, suggesting that the MAT1-2-1 product has a positive regulatory effect on the white collar genes

Evidence that the Ceratobasidium-like white-thread blight and black rot fungal pathogens from persimmon and tea crops in the Brazilian Atlantic Forest agroecosystem are two distinct phylospecies

The white-thread blight and black rot (WTBR) caused by basidiomycetous fungi of the genus Ceratobasidium is emerging as an important plant disease in Brazil, particularly for crop species in the Ericales such as persimmon (Diospyros kaki) and tea (Camellia sinensis). However, the species identity of the fungal pathogen associated with either of these hosts is still unclear. In this work, we used sequence variation in the internal transcribed spacer regions, including the 5.8S coding region of rDNA (ITS-5.8S rDNA), to determine the phylogenetic placement of the local white-thread-blight-associated populations of Ceratobasidium sp. from persimmon and tea, in relation to Ceratobasidium species already described world-wide. The two sister populations of Ceratobasidium sp. from persimmon and tea in the Brazilian Atlantic Forest agroecosystem most likely represent distinct species within Ceratobasidium and are also distinct from C. noxium, the etiological agent of the first description of white-thread blight disease that was reported on coffee in India. The intraspecific variation for the two Ceratobasidium sp. populations was also analyzed using three mitochondrial genes (ATP6, nad1 and nad2). As reported for other fungi, variation in nuclear and mitochondrial DNA was incongruent. Despite distinct variability in the ITS-rDNA region these two populations shared similar mitochondrial DNA haplotypes

Identification of Fusarium species by isozyme analysis

Cellulose-acetate electrophoresis (CAE) was used to investigate isozyme polymorphisms among different isolates of Fusarium cerealis, F. culmorum, F. graminearum and F. pseudograminearum. After initial testing of 18 enzymes in three buffer systems for activity and resolution of bands, 12 proved to be appropriate for analysis of the full sample set. Comparing the different electrophoretic types (ETs), adenylate kinase (AK), NADP dependent glutamate dehydrogenase (NADP GDH), peptidase B (PEP B), peptidase D (PEP D) and phosphoglucomutase (PGM) proved to be diagnostic for at least one species examined. However, only PEP D was useful alone as a marker to distinguish the four taxa studied providing a rapid and simple CAE based diagnostic protocol

Characterization of Parental Traits in Somatic Fusion Progeny of Phytophthora infestans and Phytophthora nicotianae

Species hybrids were created via fusion of zoospores of two morpho­logically distinct species, P. infestans and P. nicotianae. Sixteen putative hybrid isolates were recovered that expressed differential drug resistance of each parent. Repetitive DNA of P. nicotiane was detected readily in all of these isolates by hybridization with the species-specific DNA probe, pPP33A. DNA of P. infestans was detected in only two putative hybrid isolates using PCR and primer pair ITS3 and PINF2. The two true hybrids were more similar to P. nicotianae than to P. infestans on the basis of pathogenic, morphological and molecular evidence. Additionally, hybrids expressed modified host ranges compared to parental species. Fusion of zoospores or hyphae may contribute to formation of such hybrids, particularly in the case of heterothallic species in which the joint occurrence of compatible mating types is rare. Zoospore fusion may prove useful as a tool to study hybridization, pathogenesis, and sources of natural diversity of species

Identification of new molecular hallmarks for YSAPK MAPKs: application for cloning strategies in different fungal filamentous species

Based on multiple sequence alignment of mitogen activated protein kinase (MAPK) genes from 28 fungal species, we could identify twelve new hallmark sequences, specific to YSAPK (yeast and fungal stress activated protein kinases) MAPK subgroup within fungal MAPKs. Six of the motifs (I-a, I-b, IV-b, V-b, X-b1 and X-b2) showed especially high degree of specificity. Two of these six motifs, I-b (SA[RK]DQLT) and IV-b (F[IL]SPLED[IV]) were specific within eukaryotic proteins, too. The other type of motifs contained not only YSAPK specific residues but residue(s) conserved on fungal MAPK and all MAPK levels. From the viewpoint of functional role, YSAPK motif VII-a (IL[VI]NENCDL) coincided with a loop spanning 7b-8b sheets in human p38a and ERK2 MAPK proteins (consensus indicated by bold face letters). This motif was shown to be involved in interaction with L-x-L type docking motifs of activators (MAPK kinases) and transcription factors. A fungal MAPK specific signal sequence in protein kinase subdomain IX-b (AE[ML][LVI]xG[KR]PxFxG[KR]D) was also described. A subgroup specific nested PCR-based cloning approach was developed to amplify YSAPK sequences in different filamentous fungal species based on motifs I, I-b, VIII and X-b3 as primers. Putative YSAPK MAPK amplicons obtained by this approach were identified by (i) the presence of newly characterized YSAPK specific motifs and (ii) alignments to known YSAPK MAPK genes. A neighbor-joining phylogenetic tree constructed by comparing 50 fungal MAPK sequences clearly demonstrated a dichotomic origin of fungal MAPKs and separated YSAPK MAPKs from the other two fungal MAPK subgroups. Comparing the number and distribution of known MAPK genes in Fusarium and other species it is tempting to speculate that this triple structural diversity of MAPKs, at least in filamentous species is a general phenomenon. Phylogenetic analysis revealed no separation of phytopathogenic species. Blumeria graminis, Alternaria brassicicola and Aspergillus species, however were separated from the majority of filamentous Ascomycetes in all of the three MAPK subgroups

Burden of comorbid conditions in children and young people with juvenile idiopathic arthritis: a collaborative analysis of 3 JIA registries

OBJECTIVES: Burden of comorbidities are largely unknown in JIA. From 2000, national and international patient registries were established to monitor biologic treatment, disease activity and adverse events in patients with JIA. The aim of this analysis was to investigate in parallel, for the first time, three of the largest JIA registries in Europe/internationally—UK JIA Biologic Registers (BCRD/BSPAR-ETN), German biologic registers (BiKeR/JuMBO), multinational Pharmachild—to quantify the occurrence of selected comorbidities in patients with JIA. METHODS: Information on which data the registers collect were compared. Patient characteristics and levels of comorbidity were presented, focussing on four key conditions: uveitis, MAS, varicella, and history of tuberculosis. Incidence rates of these on MTX/biologic therapy were determined. RESULTS: 8066 patients were registered into the three JIA registers with similar history of the four comorbidities across the studies; however, varicella vaccination coverage was higher in Germany (56%) vs UK/Pharmachild (16%/13%). At final follow-up, prevalence of varicella infection was lower in Germany (15%) vs UK/Pharmachild (37%/50%). Prevalence of TB (0.1–1.8%) and uveitis (15–19%) was similar across all registers. The proportion of systemic-JIA patients who ever had MAS was lower in Germany (6%) vs UK (15%) and Pharmachild (17%). CONCLUSION: This analysis is the first and largest to investigate the occurrence of four important comorbidities in three JIA registries in Europe and the role of anti-rheumatic drugs. Combined, these three registries represent one of the biggest collection of cases of JIA worldwide and offer a unique setting for future JIA outcome studies