Orchids require symbiotic fungi and/or specific conditions to germinate. Asymbiotic techniques have been shown successful for orchid germination. In Chile, Chloraea include many endemic, and potentially ornamental, terrestrial orchid species. In this study, individuals of Chloraea crispa, C. gavilu and C. virescens were manually autopollinated. The resulting capsules were sterilized and seeds were aseptically obtained. We evaluated asymbiotic germination in: Agar Water (AW), Knudson C (KC), Banana Culture Media (CMB), Tomato Culture Media (CMT), Malmgren Modified (MM), Murashige and Skoog (MS), and MS modified (MS1/2). Seeds were incubated in the dark at 24±1 ºC for 2 weeks. Then they were put in 16/8 h light/dark cycles for 14 weeks. We registered germination and embryo development in the different culture media. After 8 weeks, all tested Chloraea species germinated in most culture media. After 16 weeks, embryos in MM showed an evident shoot. In AW only the pre-germination stage was achieved. In C. gavilu germination was over 90% in MM, CMT, and CMB. In KC, a lower germination was obtained in all tested species compared to the other culture media, except for AW. Asymbiotic germination of endemic Chilean orchids could be a viable strategy for their conservation and propagation as ornamental species.

Almost all orchid species depend on association with fungal partners. These fungal species facilitate orchid seed germination and promote growth and possibly stress tolerance in adult plants, both in the field and laboratory conditions. Codonorchis lessonii is a terrestrial orchid, endemic to southern Chile and Argentina with a currently unknown conservation status. Previous studies have reported Rhizoctonia-like fungi associated with C. lessonii in Argentina, but their fungal partners in Chilean populations are unknown. This study aims to characterize and isolate the mycorrhizal fungi associated with adult individuals of C. lessonii from three different populations in Central-South Chile. Root sections showing pelotons were used for the isolation and identification of the fungal taxa. Radial fungal growth was measured for all mycelial isolates. The presence of binucleate cells placed all isolates within the families Ceratobasidiaceae and Tulasnellaceae and the identification was confirmed by molecular analysis. Fungal isolates belonging to Ceratobasidiaceae grew at a higher rate than those from Tulasnellaceae. Phylogenetic analyses showed that different fungal partners associate with this orchid, suggesting relatively low specificity. The isolation and identification of the fungal partners of C. lessonii could help understand its ecology and contribute in future restoration and propagation initiatives for the species.

Fungal polysaccharides possess a broad biological activity, including cytotoxic and antioxidant activities. This work aimed to evaluate the cytotoxic and antioxidant activity of the acidic polysaccharides of Phylloporia boldo strain (named PBAP40). Cytotoxic activity of polysaccharide was evaluated determining the viability of three tumor cell lines by MTT assay. The effect of acidic polysaccharide on the cell cycle of HL-60 cell line was evaluated by flow cytometry, and the antioxidant activity was determined by DPPH and ABTS assays. PBAP40 showed cytotoxic effects in tumor cell lines. Results suggest that P. boldo acidic polysaccharides arrested tumor cells in the cell cycle Sub G1 phase. The acidic polysaccharides of PBAP40 strain were not cytotoxic for the non-tumor cell line. PBAP40 also showed excellent antioxidant activity.The FT-IR analysis of the acidic polysaccharides indicated the presence of glucans bearing α-and β-type glycosidic bonds.