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Macroalgae of the genera Fucus, Ulva , and Enteromorpha are typically abundant in estuaries. Endophytic fungi may have beneficial effects on the hosts affecting their ability to cope with stress. They are also a source of biologically active compounds. However, little is known about the endophytic fungi that colonize these macroalgae. Endophytic isolates were obtained from macroalgae from various sites in the estuary Ria de Aveiro (Portugal), as well as from saline water and sponges. Six Acremonium -like species could not be affiliated to any known species. Phylogenetic analyses based on internal transcribed spacer (ITS) region of the ribosomal DNA and β-tubulin ( tub2 ) and actin ( act1 ) genes placed these species in the genera Emericellopsis and Parasarocladium , but distinct from all currently known species. Although sharing morphological characteristics with the most closely related species, these genera differ in micromorphological and molecular characters. Thus, three novel species of Emericellopsis ( E. cladophorae , sp. nov., E. enteromorphae , sp. nov., and E. phycophila , sp. nov.) and three novel species of Parasarocladium ( P. aestuarinum , sp. nov., P. alavariense , sp. nov., and P. fusiforme , sp. nov.) are proposed.

Lasiodiplodia theobromae is a fungal plant pathogen that has been associated with Botryosphaeria dieback of grapevine. Despite several studies on L. theobromae , until now the production of secondary metabolites by strains isolated from grapevines has not been reported. The ability of two strains of L. theobromae isolated from grapevine to produce lipophilic metabolites was studied. Although many typical compounds of low molecular weight were identified from the crude extracts of both strains (e.g., lasiolactols, substituted 2-dihydrofuranones, melleins, jasmonic acid, 3-indolcarboxylic acid, botryodiplodins), (2 R /2 S ,3 S ,4 S )-3- epi -botryodiplodin was isolated for the first time as a natural compound. Furthermore, a comparative study of metabolite production was conducted at 25 and 37 C to understand temperature effects on metabolite profiles. Some metabolites were produced only by one strain (e.g., (3 S ,4 S )-4-acetyl-3-methyl-2-dihydrofuranone produced by LA-SOL3) and others only at a specific temperature (e.g., jasmonic acid at 25 C, botryodiplodins at 37 C). Phytotoxicity and cytotoxicity of pure compounds were evaluated to clarify the influence of lipophilic metabolites on the biological activities of culture filtrates of both strains. The most toxic compound for Vero and 3T3 cells was (2 R /2 S ,3 S ,4 S )-3- epi -botryodiplodin.

Lasiodiplodia theobromae is a phytopathogenic fungus that causes diseases not only in a broad number of plant hosts but also occasionally in humans. The capacity of L. theobromae to produce bioactive metabolites at 25 C (environmental mean temperature) and at 37 C (body mean temperature) was investigated. Two strains, CAA019 and CBS339.90, isolated respectively from a coconut tree and a human patient were characterized. The phytotoxicity and cytotoxicity (on mammalian cells) of the secretomes of both strains of L. theobromae were investigated. Also, phytotoxicity and cytotoxicity of pure compounds were evaluated. The phytotoxicity of the secretome of strain CAA019 was higher than the phytotoxicity of the secretome of strain CBS339.90 at 25 C. However, the phytotoxicity for both strains decreased when they were grown at 37 C. Only the secretome of strain CBS339.90 grown at 37 C induced up to 90% Vero and 3T3 cell mortality. This supports the presence of different metabolites in the secretome of strains CAA019 and CBS339.90. Metabolites typical of L. theobromae were isolated and identified from organic extracts of the secretome of both strains (e.g., 3-indolecarboxylic acid, jasmonic acid, lasiodiplodin, four substituted 2-dihydrofuranones, two melleins, and cyclo-(Trp-Ala)). Also, metabolites such as scytalone, not previously reported for this species, were isolated and identified. Metabolite production is affected by strain and temperature. In fact, some metabolites are strain specific (e.g., lasiodiplodin) and some metabolites are temperature specific (e.g., jasmonic acid). Although more strains should be characterized, it may be anticipated that temperature tuning of secondary-metabolite production emerges as a putative contributing factor in the modulation of L. theobromae pathogenicity towards plants, and also towards mammalian cells.