Phylogenetic and chemotaxonomic resolution of the genus Annulohypoxylon(Xylariaceae)including four new species

This study deals with an extensive evaluation focusing on phylogenetic and chemotaxonomic infrageneric relationships of the genus Annulohypoxylon(Xylariaceae,Ascomycota),whose species are ubiquitously associated with seed plants as endophytes and saprotrophs in all forested areas of the world.Using evidence from phylogenetic,chemotaxonomic and morphological data,various varieties within the genus are raised to species level,leading to the new combinations for A.areolatum(:A.bovei var.microsporum),A.macrosporum(:A.leptascum var.macrosporum),and A.microdiscum(:A.moriforme var.microdiscum).Annulohypoxylon substygium nom.nov.is applied for A.stygium var.annulatumand the four new tropical and subtropical species A.massivum,A.violaceopigmentum,A.viridistratum and A.yungensis are introduced.Furthermore,A.leucadendri is excluded from the genus as its morphological characters disagree with the generic concept,the recently erected A.palmicola is synonymized with A.leptascum and A.austrobahiense has been reassigned to the genus Hypoxylon.In addition,the key taxa A.annulatum and A.truncatum have been reinvestigated and epitypes have been defined.For the first time,a dichotomous key to the genus is provided.A phylogenetic b-tubulin tree in conjunction with stromatal HPLC profiles clearly shows that Annulohypoxylon comprises two distinct lineages.The A.cohaerens/multiforme group might eventually warrant segregation into a new genus as further molecular data become available.

Identification of fungal fossils and novel azaphilone pigments in ancient carbonised specimens of Hypoxylon fragiforme from forest soils of Chatillon-sur-Seine(Burgundy)

Fungal stromata were recently discovered in association with charcoal and burnt soil aggregates during an archaeological survey in the Chaˆtillon-sur-Seine forest massif.The wood and soil in the samples were dated to the medieval period(between 738 and 1411 AD).Light microscopy and scanning electron microscopy revealed that a few of the stromatal fragments still contained ascospores.Their macromorphological characters were described and secondary metabolite profiles were generated using high performance liquid chromatography with diode array and mass spectrometric detection(HPLC–DAD/MS).The combination of these two data lines then allowed species identification.Most of the fragments were assigned to Hypoxylon fragiforme,the type species of the Hypoxylaceae(Xylariales).Two further species whose stromata grew on the fossil charcoal could be tentatively identified as Jackrogersella cohaerens and(more tentatively)as Hypoxylon vogesiacum.These three species are still commonly encountered in the forests of Central Europe today.Furthermore,the HPLC-HRMS data of H.fragiforme suggested the presence of unknown azaphilone dimers and of further new pigments.These archaeological compounds were compared to fresh stromata of H.fragiforme collected in Germany and subjected to the same analytical protocol.While the major components in both samples were identified as the known mitorubrin type azaphilones and orsellinic acid,the chemical structures of seven novel complex azaphilone pigments,for which we propose the trivial names rutilins C-D and fragirubrins A-E,were elucidated using spectral methods(NMR and CD spectroscopy,high resolution mass spectrometry).It appears that these pigments had indeed persisted for millennia in the fossil stromata.

The Group A Streptococcus Interleukin-8 Protease SpyCEP Promotes Bacterial Intracellular Survival by Evasion of Autophagy

Autophagy serves an innate immune function in defending the host against invading bacteria,including group A Streptococcus(GAS).Autophagy is regulated by numerous host proteins,including the endogenous negative regulator calpain,a cytosolic protease.Globally disseminated serotypeM1T1 GAS strains associated with high invasive disease potential express numerous virulence factors and resist autophagic clearance.Upon in vitro infection of human epithelial cell lines with representative wild-type GAS M1T1 strain 5448(M1.5448),we observed increased calpain activation linked to a specific GAS virulence factor,the interleukin-8 protease SpyCEP.Calpain activation inhibited autophagy and decreased capture of cytosolic GAS in autophagosomes.In contrast,the serotype M6 GAS strain JRS4(M6.JRS4),which is highly susceptible to host autophagy-mediated killing,expresses low levels of SpyCEP and does not activate calpain.Overexpression of SpyCEP in M6.JRS4 stimulated calpain activation,inhibited autophagy,and significantly decreased bacterial capture in autophagosomes.These paired loss-and gain-of-function studies reveal a novel role for the bacterial protease SpyCEP in enabling GAS M1 evasion of autophagy and host innate immune clearance.