SUMMARYThe genusTrichodermacontains fungi with high relevance for humans, with applications in enzyme production for plant cell wall degradation and use in biocontrol. Here, we provide a broad, comprehensive overview of the genomic content of these species for “hot topic” research aspects, including CAZymes, transport, transcription factors, and development, along with a detailed analysis and annotation of less-studied topics, such as signal transduction, genome integrity, chromatin, photobiology, or lipid, sulfur, and nitrogen metabolism inT. reesei,T. atroviride, andT. virens, and we open up new perspectives to those topics discussed previously. In total, we covered more than 2,000 of the predicted 9,000 to 11,000 genes of eachTrichodermaspecies discussed, which is >20% of the respective gene content. Additionally, we considered available transcriptome data for the annotated genes. Highlights of our analyses include overall carbohydrate cleavage preferences due to the different genomic contents and regulation of the respective genes. We found light regulation of many sulfur metabolic genes. Additionally, a new Golgi 1,2-mannosidase likely involved inN-linked glycosylation was detected, as were indications for the ability ofTrichodermaspp. to generate hybrid galactose-containingN-linked glycans. The genomic inventory of effector proteins revealed numerous compounds unique toTrichoderma, and these warrant further investigation. We found interesting expansions in theTrichodermagenus in several signaling pathways, such as G-protein-coupled receptors, RAS GTPases, and casein kinases. A particularly interesting feature absolutely unique toT. atrovirideis the duplication of the alternative sulfur amino acid synthesis pathway.

No abstract

No abstract

The industrially important cellulolytic filamentous fungus Trichoderma reesei is the anamorph of the pantropical ascomycete Hypocrea jecorina. H. jecorina CBS999.97 strain undergoes a heterothallic reproductive cycle, and the mating yields fertilized perithecia imbedded in stromata. Asci in the perithecia contain 16 linearly arranged ascospores. Here, we investigated H. jecorina sexual development under different light regimes, and found that visible light was dispensable for sexual development (stroma formation and ascospore discharge). By contrast, constant illumination inhibited stroma formation, and an interruption of the darkness facilitated timely stroma formation in a 12 h/12 h light-dark photoperiod. The results of genetic analyses further revealed that H. jecorina blue-light photoreceptors (BLR1, BLR2) and the photoadaptation protein ENV1 were not essential for sexual development in general. BLR1, BLR2 and ENV1 are orthologues of the conserved Neurospora crassa WC-1, WC-2 and VVD, respectively. Moreover, BLR1 and BLR2 mediate both positive and negative light-dependent regulation on sexual development, whereas ENV1 is required for dampening the light-dependent inhibitory effect in response to changes in illumination. Comparative genome-wide microarray analysis demonstrated an overview of light-dependent gene expression versus sexual potency in CBS999.97 (MAT1–2) haploid cells. Constant illumination promotes abundant asexual conidiation and high levels of hpp1 transcripts. hpp1 encodes a h (hybrid)-type propheromone that exhibits features of both yeast a and a pheromone precursors. Deletion of hpp1 could rescue stroma formation but not ascospore generation under constant illumination. We inferred that the HPP1-dependent pheromone signaling system might directly prevent stroma formation or simply disallow the haploid cells to acquire sexual potency due to abundant asexual conidiation upon constant illumination.

We present an automated method for measuring the sagittal vertical axis (SVA) from lateral radiography of whole spine using a convolutional neural network for keypoint detection (ResUNet) with our improved localization method. The algorithm is robust to various clinical conditions, such as degenerative changes or deformities. The ResUNet was trained and evaluated on 990 standing lateral radiographs taken at Chang Gung Memorial Hospital, Linkou and performs SVA measurement with median absolute error of 1.183 ± 0.166 mm. The 5-mm detection rate of the C7 body and the sacrum are 91% and 87%, respectively. The SVA calculation takes approximately 0.2 s per image. The intra-class correlation coefficient of the SVA estimates between the algorithm and physicians of different years of experience ranges from 0.946 to 0.993, indicating an excellent consistency. The superior performance of the proposed method and its high consistency with physicians proved its usefulness for automatic measurement of SVA in clinical settings.

SUMMARYStriatin family proteins are key regulators in signalling pathways in fungi and animals. These scaffold proteins contain four conserved domains: a caveolin-binding domain, a coiled-coil motif and a calmodulin-binding domain at the N-terminus, and a WDrepeat domain at the C-terminus. Fungal striatin orthologues are associated with sexual development, hyphal growth and plant pathogenesis. In Fusarium verticillioides, the striatin orthologue Fsr1 promotes virulence in the maize stalk. The relationship between fungal striatins and pathogenicity remains largely unexplored. In this study, we demonstrate that the Colletotrichum graminicola striatin orthologue Str1 is required for full stalk rot and leaf blight virulence in maize. Pathogenicity assays show that the striatin mutant strain (Dstr1) produces functional appressoria, but infection and colonization are attenuated. Additional phenotypes of the Dstr1 mutant include reduced radial growth and compromised hyphal fusion. In comparison with the wild-type, Dstr1 also shows a defect in sexual development and produces fewer and shorter conidia. Together with the fact that F. verticillioides fsr1 can complement Dstr1, our results indicate that C. graminicola Str1 shares five phenotypes with striatin orthologues in other fungal species: hyphal growth, hyphal fusion, conidiation, sexual development and virulence. We propose that fungal striatins, like mammalian striatins, act as scaffolding molecules that cross-link multiple signal transduction pathways.

Appressoria are essential penetration structures for many phytopathogenic fungi. Here F-actin localization dynamics were documented during appressorium formation in vitro and in planta in Colletotrichum graminicola Four discernible stages of dynamic F-actin distribution occurring in a programmed order were documented from differentiation of appressoria to formation of penetration pores: (stage A) from germ tube enlargement to complete expansion of the appressorium; (stage S) septation occurs; (stage L) a long period of low F-actin activity; (stage P) the penetration pore forms. The F-actin subcellular localization corresponded to each stage. A distinct redistribution of actin cables occurred at the transition from stage A to stage S. The in planta assays revealed that F-actin also assembled in invasive hyphae and that actin cables might play an essential role for penetration-peg development. The F-actin localization distribution may be used as a subcellular marker to define the developmental stages during appressorium formation.

Summary Monilinia fructicola (G. Winter) Honey is a devastating pathogen on Rosaceae which causes blossom blight and fruit rot. Only a few studies related to the plant–pathogen interaction have been published and there is limited knowledge on the relationship between oxidative stress and successful infection in M. fructicola. In this study, we cloned and characterized a redox‐responsive transcription factor MFAP1, a YAP1 homologue. MfAP1‐silenced strains were generated by polyethylene glycol‐mediated protoplast transformation or Agrobacterium T‐DNA‐mediated transformation. Pathogenicity assay demonstrated that MfAP1‐silenced strains caused smaller lesions on rose and peach petals. Transformants carrying extra copies of MfAP1, driven by the native promoter, were generated for MfAP1 overexpression. Interestingly, MfAP1‐overexpressing strains also caused smaller lesions on rose petals. Strains carrying two copies of MfAP1 accumulated reactive oxygen species (ROS) at higher levels and exhibited delayed accumulation of MfAP1 transcripts compared with the wild‐type during pathogenesis. By the analysis of ROS production and the expression patterns of redox‐ and virulence‐related genes in the wild‐type strain and an MfAP1‐overexpressing strain, we found that the M. fructicola wild‐type strain responded to oxidative stress at the infection site, activated the expression of MfAP1 and up‐regulated the genes required for ROS detoxification and fungal virulence. In contrast, MfAP1 expression in the MfAP1‐overexpressing strain was suppressed after the induction of a strong oxidative burst at the infection site, altering the expression of ROS detoxification and virulence‐related genes. Our results highlight the importance of MfAP1 and ROS accumulation in the successful infection of M. fructicola.