Primary Characterization of a Life-Cycle Mutant akasusabi of the Red Alga Neopyropia yezoensis

Gametophyte-to-sporophyte transition in the haploid-diploid life cycle depends on fertilization of male and female gametes. We describe here a mutant of the marine red seaweed Neopyropia yezoensis, designated akasusabi (aks), where the gametophyte-to-sporophyte transition occurs independently of fertilization. Although conchocelis filaments were produced from carpospores, severe defects in the maturation of carposporangia via mitosis to generate conchospores were observed. In the aks mutant, however, somatic cells of gametophytic thalli were able to produce conchocelis filaments without fertilization. Thus, apogamy occurs in aks. In addition, aks was highly sensitive to wounding that promotes both asexual and apogamous reproductive responses by producing spores, which develop either into blades or conchocelis filaments, indicating that aks responds to wounding by enhanced reproduction. These findings indicated that the aks mutation enables the transformation of vegetative cells to carpospores to produce sporophytes by apogamy and wound-inducible life cycle trade-off, stimulating a reset of the timing of reproduction during the life cycle. Therefore, AKS is involved in regulations of the gametophyte-to-sporophyte transition and asexual spore production in N. yezoensis.

Ecology of Diaporthe eres, the causal agent of hazelnut defects

Diaporthe eres has been recently reported as the causal agent of hazelnut defects, with characteristic brown spots on the kernels surface and internal fruit discoloration. Knowledge regarding the ecology of this fungus is poor but, is critical to support a rationale and effective hazelnut crop protection strategy. Therefore, a study was performed to describe and model the effect of different abiotic factors such as temperature (T, 5–35°C, step 5°C) and water activity (aw 0.83–0.99, step 0.03) regimes on D. eres mycelial growth, pycnidial conidiomata development and asexual spore production during a 60-day incubation period. Alpha conidia germination was tested in the same T range and at different relative humidities (RH = 94, 97 and 100%) over 48 h incubation period. Fungal growth was observed from the first visual observation; regarding pycnidia and cirrhi, their development started after 8 and 19 days of incubation, respectively and increased over time. The optimum T for growth was 20–25°C and for pycnidia and cirrhi development was 30°C; aw ≥ 0.98 was optimal for the tested steps of the fungal cycle. The best condition for conidial germination of D. eres was at 25°C with RH = 100%. Quantitative data obtained were fitted using non- linear regression functions (Bete, logistic and polynomial), which provided a very good fit of the biological process (R2 = 0.793–0.987). These functions could be the basis for the development of a predictive model for the infection of D. eres of hazelnuts.

The Potential of Lipolytic Filamentous Fungi Isolated from Landfill Soil as Poly-β-Hydroxybutirate (PHB) Bioplastic Degrader

The present study was investigated the potential of lipolytic fungi (molds) isolated from landfill soil in degrading Poly-β-hydroxybutyrate (PHB). Screening of PHB-degrading lipolytic molds was done in two stages, such as screening of lipolytic molds which was identified by the formation of orange fluorescent halos around the colony on rhodamine B agar medium and the degradation PHB ability test was identified by the formation of clear zone around colony on PHB emulsion medium. Characterization of isolates was done based on phenotypic characters and the identification was done by numerical-phenetic analysis. Three lipolytic mold isolates that have ability in degrading PHB bioplastic i.e isolate KC1, KE1 and KE6. These molds have asexual spore form conidia, foot cell, septate hyphae, unbranched conidiophore, and spore mass located at the apex of phialid. The identification results showed that isolate KC1 is identic to Aspergillus terreus, KE1 is identic to Aspergillus niger and KE6 is identic to Aspergillus fumigatus.

A Small Horizontally Transferred Gene Cluster Contributes to the Sporulation of Alternaria alternata

Horizontal gene transfer (HGT) has been identified as an important source of genomic innovation in fungi. However, how HGT drove the evolution of Alternaria alternata, a necrotrophic fungus which can be ubiquitously isolated from soil and various plants and decaying plant materials is largely known. In this study, we identified 12 protein-encoding genes that are likely acquired from lineages outside Pezizomycotina. Phylogenetic trees and approximately unbiased comparative topology tests strongly supported the evolutionary origin of these genes. According to their predicted functions, these HGT candidates are involved in nitrogen and carbohydrate metabolism. Especially, five genes of them were likely transferred as a physically linked cluster from Tremellales (Basidiomycota). Functionally knocking out the five-gene cluster in an A. alternata isolate causing citrus brown spot resulted in an 80% decrease in asexual spore production in the deletion mutant. We further knocked out each of these five genes in this cluster and the resultant single-gene deletion mutants exhibited a various degree of reduction in spore production. Except for conidiation, functions of these genes associated with vegetative growth, stress tolerance, and virulence are very limited. Our results provide new evidence that HGT has played important roles over the course of the evolution of filamentous fungi.

Label-Free Quantitative Proteomics of Lysine Acetylome Identifies Substrates of Gcn5 in Magnaporthe oryzae Autophagy and Epigenetic Regulation

ABSTRACT The rice blast fungus Magnaporthe oryzae poses a great threat to global food security. During its conidiation (asexual spore formation) and appressorium (infecting structure) formation, autophagy is induced, serving glycogen breakdown or programmed cell death function, both essential for M. oryzae pathogenicity. Recently, we identified an M. oryzae histone acetyltransferase (HAT) Gcn5 as a key regulator in phototropic induction of autophagy and asexual spore formation while serving a cellular function other than autophagy induction during M. oryzae infection. To further understand the regulatory mechanism of Gcn5 on M. oryzae pathogenicity, we set out to identify more Gcn5 substrates by comparative acetylome between the wild-type (WT) and GCN5 overexpression (OX) mutant and between OX mutant and GCN5 deletion (knockout [KO]) mutant. Our results showed that Gcn5 regulates autophagy induction and other important aspects of fungal pathogenicity, including energy metabolism, stress response, cell toxicity and death, likely via both epigenetic regulation (histone acetylation) and posttranslational modification (nonhistone protein acetylation). IMPORTANCE Gcn5 is a histone acetyltransferase that was previously shown to regulate phototropic and starvation-induced autophagy in the rice blast fungus Magnaporthe oryzae, likely via modification on autophagy protein Atg7. In this study, we identified more potential substrates of Gcn5-mediated acetylation by quantitative and comparative acetylome analyses. By epifluorescence microscopy and biochemistry experiments, we verified that Gcn5 may regulate autophagy induction at both the epigenetic and posttranslational levels and regulate autophagic degradation of a critical metabolic enzyme pyruvate kinase (Pk) likely via acetylation. Overall, our findings reveal comprehensive posttranslational modification executed by Gcn5, in response to various external stimuli, to synergistically promote cellular differentiation in a fungal pathogen.

Flora’s Secret Gardens

Wilhelm Hofmeister established the unity of the Plant Kingdom through the discovery of Alternation of Generations. In both cryptogams and flowering plants a diploid asexual stage, or sporophyte, alternates with a haploid sexual stage. Thus the flower is not the true sexual stage, but rather the asexual spore-producing stage. The main difference between ferns and roses is that the spores of the fern are visible on the undersides of the leaves, while the spores of the rose are concealed within the anthers and ovaries. These spores develop into the actual sexual stage of the spermatophyte, the male and female gametophytes, i.e. the pollen tube and the embryo sac. Hofmeister’s discovery solved of the age-old quandary over plant sex. The sexualists and the asexualists can both claim to have been correct, but it was the sexualists who freed their minds from cultural biases and glimpsed the true sexual nature of plants.

Histidine Kinase Two-Component Response Regulator Proteins Regulate Reproductive Development, Virulence, and Stress Responses of the Fungal Cereal Pathogens Cochliobolus heterostrophus and Gibberella zeae

ABSTRACT Histidine kinase (HK) phosphorelay signaling is a major mechanism by which fungi sense their environment. The maize pathogen Cochliobolus heterostrophus has 21 HK genes, 4 candidate response regulator (RR) genes (SSK1, SKN7, RIM15, REC1), and 1 gene (HPT1) encoding a histidine phosphotransfer domain protein. Because most HKs are expected to signal through RRs, these were chosen for deletion. Except for pigment and slight growth alterations for rim15 mutants, no measurable altered phenotypes were detected in rim15 or rec1 mutants. Ssk1p is required for virulence and affects fertility and proper timing of sexual development of heterothallic C. heterostrophus. Pseudothecia from crosses involving ssk1 mutants ooze masses of single ascospores, and tetrads cannot be found. Wild-type pseudothecia do not ooze. Ssk1p represses asexual spore proliferation during the sexual phase, and lack of it dampens asexual spore proliferation during vegetative growth, compared to that of the wild type. ssk1 mutants are heavily pigmented. Mutants lacking Skn7p do not display any of the above phenotypes; however, both ssk1 and skn7 mutants are hypersensitive to oxidative and osmotic stresses and ssk1 skn7 mutants are more exaggerated in their spore-type balance phenotype and more sensitive to stress than single mutants. ssk1 mutant phenotypes largely overlap hog1 mutant phenotypes, and in both types of mutant, the Hog1 target gene, MST1, is not induced. ssk1 and hog1 mutants were examined in the homothallic cereal pathogen Gibberella zeae, and pathogenic and reproductive phases of development regulated by Ssk1 and Hog1 were found to mirror, but also vary from, those of C. heterostrophus.