LFFs associating with green algae utilize polyols therefore

LFFs associating with green algae utilize polyols; therefore, we hypothesize that at least one polyol transport system is conserved in LFFs and it is required for the lichenization events that occur widely and independently in Ascomycota. Thus, the present study aimed to first evaluate phylogenetic relationships between polyol and monosaccharide transporters and then confirm the conservation of polyol transporters in ascomycetous fungi, including LFFs. On the basis of the analyses, we also speculated that there was a correlation between polyol transporters and lichenization in ascomycetous fungi, especially those associated with green algae. To determine the predicted polyol transporters, the eight genome sets of ascomycetous lichenized fungi were analyzed. Moreover, we also sequenced the whole genome of LFF Ramalina conduplicans and selected candidate polyol transporter genes. We also isolated the candidates that were expressing when R. conduplicans was cultured on a medium containing ribitol as the sole carbon source. Ramalinaceae, including R. conduplicans, is the fourth largest of the 115 GSK J4 representing lichenized fungi (Lücking et al., 2016). The genus Ramalina grows on culture media relatively quickly (Komiya and Shibata, 1971) and has been used for chemical and biological activity studies of extracts and isolated compounds (reviewed in Moreira et al., 2015). Additionally, it is distributed worldwide (Kirk et al., 2008). Some metabolites of R. conduplicans have possible industrial, pharmaceutical and agricultural uses (Oh et al., 2006, Wei et al., 2008, Vinayaka et al., 2009, Luo et al., 2010), and R. conduplicans is associated with Trebouxia spp., the most common photosynthetic partner in lichens (Honegger, 2008). Therefore, R. conduplicans could be a lichen model for studying secondary metabolites and polyol utilization. > Materials and methods
Results and discussion
Conclusion Polyol transporter genes are conserved in nearly all major ascomycetous lineages, regardless of whether they are lichen-forming (Fig. 3A and B). The maintenance of polyol transporter genes may explain why lichenization events have occurred many times in Ascomycota. However, LFFs belonging to Lecanoromycetes tend to retain multiple proteins similar to A. monospora’ Lat2. Thus, Lecanoromycetes may have duplicated polyol transporter genes for stronger associations with green algae after a lichenization events. Degenerate primers designed based on conserved domains allowed the isolation of polyol transporter-like genes from the cDNA of R. conduplicans that corresponded with the genome sequences. This suggested that further candidate genes could be collected from LFFs, such as basidiolichens and cyanolichens, photosynthetic partners, lichenicolous fungi and endolichenic microorganisms, even if their genomic data are not available. In addition, broad experiments, including those elucidating the gene expression patterns, post-translational modifications and horizontal gene transfer events, are also needed to investigate the polyol transfer systems in LFFs and their symbiotic association processes with green algae. Finally, we have begun to uncover possible connections between the evolution of diverse polyol transporters in the phylum Ascomycota and the success of lichenized fungi.
Acknowledgments We are very grateful to Dr. Hiroshi Harada (Natural History Museum and Institute, Chiba, Japan) for supporting the morphological observations and accepting our specimens. This study was supported by the Research Fellowships for Young Scientists (No. 17J03039) from the Japan Society for the Promotion of Science (JSPS), and the Fujiwara Natural History Public Interest Incorporated Foundation.
Introduction Biogenic amine histamine is synthesized from L-histidine by histidine decarboxylase (HDC). Histamine is involved in a wide variety of pathological and physiological processes including allergic reactions, inflammation, immune responses, gastric secretion, and neurotransmission. Intracellularly synthetized histamine is stored in cytoplasmic granules of mast cells and basophils, two of the major producers of histamine. Basophilic leukemia (RBL)-2H3 cells, a tumor analog of rat mast cells, is considered to be a convenient model to study immunoglobulin E (IgE)-mediated degranulation of mast cells, because they express the IgE receptor, store mediators in granules, and release mediators, such as histamine, β-hexosaminidase, and serotonin, after crosslinking IgE (Maeyama et al., 1986, Gilfillan and Beaven, 2012).