A Nodulisporium species (designated Ti-13) was isolated as an endophyte from Cassia fistula. The fungus produces a spectrum of volatile organic compounds (VOCs) that includes ethanol, acetaldehyde and 1,8-cineole as major components. Initial observations of the fungal isolate suggested that reversible attenuation of the organism via removal from the host and successive transfers in pure culture resulted in a 50 % decrease in cineole production unrelated to an overall alteration in fungal growth. A compound (CPM 1 ) was obtained from Betula pendula (silver birch) that increases the production of 1,8-cineole by an attenuated Ti-13 strain to its original level, as measured by a novel bioassay method employing a 1,8-cineole-sensitive fungus (Sclerotinia sclerotiorum). The host plant produces similar compounds possessing this activity. Bioactivity assays with structurally similar compounds such as ferulic acid and gallic acid suggested that the CPM 1 does not act as a simple precursor to the biosynthesis of 1,8-cineole. NMR spectroscopy and HPLC-ES-MS indicated that the CPM 1 is a para-substituted benzene with alkyl and carboxyl substituents. The VOCs of Ti-13, especially 1,8-cineole, have potential applications in the industrial, fuel and medical fields.
A novel endophyte designated Collophora aceris, was obtained from stem tissues of Douglas Maple (Acer glabrum var. douglasii) in a Pacific Northwest temperate rainforest. Colonies were slow growing, white, creamy, moist, and translucent to opaque on potato dextrose agar and other media with few aerial hyphae. It also produced solid, dark sclerotia (200-400 μm) on oatmeal agar and no evidence of pseudopycnidia as per other Collophora spp. Conidia were rod-like in the size ranging from 2.2-8.4 × 0.8-1.8 μm and produced holoblastically on conidiogenous cells by budding with no collarette at the budding site. Phylogenetic analyses, based on 18S rDNA sequence data, showed that C. aceris possessed 99 % similarity to other Collophora spp. However, ITS-5.8S rDNA sequence data indicated that the organism was potentially related to Allantophomopsis spp. Finally, combined morphological, physiological, and molecular genetics data indicated that this organism is most like Collophora spp. but it is distinctly unique when compared to all other fungi in this group. It is to be noted that this is the first report of any member of this genus existing as an endophyte. This fungus makes a wide spectrum antimycotic agent (Collophorin) with biological activity against such pathogenic fungi as Pythium ultimum, Phytophthora cinnamomi, Phytophthora palmivora, and Rhizoctonia solani. Collophorin was purified to homogeneity and shown to have a unique mass of 120.0639, an empirical formula of C8H8O1, and UV absorption bands at 260 and 378 nm. This work also indicates that C. aceris possesses the biological potential to provide protection of its host against an array of common plant pathogens.
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