Four strains representing three novel anamorphic yeast species were isolated from the external surface of sugarcane leaves (DMKU-RK254(T)), corn leaves (DMKU-RK548(T)), bean leaves (K129) in Thailand and hengchun pencilwood leaves (TrB1-1(T)) in Taiwan. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, the sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene, the internal transcribed spacer (ITS) region, the actin gene (ACT1) and the elongation factor 2 gene (EF2), the four strains were determined to represent novel Yamadazyma species although formation of ascospores was not observed. Strain DMKU-RK254(T) was determined to be related to Candida diddensiae, Candida naeodendra and Candida kanchanaburiensis but with 1.8, 1.8 and 2.0 % nucleotide substitutions in the D1/D2 region of the LSU rRNA gene, respectively. It was assigned to Yamadazyma siamensis sp. nov. (type strain DMKU-RK254(T) = BCC 50730(T) = NBRC 108901(T) = CBS 12573(T)). The sequences of the D1/D2 region of the LSU rRNA gene, the ITS region, ACT1 gene and EF2 gene of two strains (DMKU-RK548(T) and K129) were identical but differed from that of strain TrB1-1(T) by 0.6, 1.0, 3.3 and 5.9 % nucleotide substitutions, respectively. Therefore, the two strains (DMKU-RK548(T) and K129) and strain TrB1-1(T) were assigned to be two separate species. The closest species in terms of pairwise sequences similarity of the D1/D2 region to the two novel species was Yamadazyma philogaea but with 1.1-1.7 % nucleotide substitutions. The two strains (DMKU-RK548(T) and K129) were assigned to Yamadazyma phyllophila sp. nov. (type strain DMKU-RK548(T) = BCC 50736(T) = NBRC 108906(T) = CBS 12572(T)) and the strain TrB1-1(T) was named Yamadazyma paraphyllophila sp. nov. (type strain TrB1-1(T) = BCRC 23030(T) = CCTCC AY 204005(T) = CBS 9928(T)).
Spider silks are protein-based fibers that are incorporated into webs with the unique combination of high mechanical toughness and resistance to microbial degradation. While spiders are undoubtedly exposed to saprophytic microorganisms in their native habitats, such as the forest understory and bush, their silks have rarely been observed to decompose in either field or laboratory studies. We performed cross-streaking assays using silk from three spider species and four bacterial strains and found no inhibition zones, indicating the absence of antibacterial properties. We also cultured all bacteria directly upon silk in Luria-Bertani broth (full nutrients), Phosphate-buffered saline (no nutrients) and nitrogen-free glucose broth (full nutrients, no nitrogen), and found bacteria grew readily on LB broth but not in PBS or NFG buffer. Our results indicated that spider silk's resistance to bacterial degradation is likely due to bacteriostatic, rather than antibacterial, mechanisms, as nitrogen is made unavailable.
Among many isolates that resulted from four independent surveys of yeasts associated with plants in Brazil, the USA, Portugal and Taiwan, we have characterized eighteen basidiomycetous strains, two of which were conspecific with the type strain of Rhodotorula acheniorum, whereas the remaining sixteen isolates appeared not to correspond to any previously described species. Microsatellite-PCR fingerprinting with primers M13 and (GTG)5 confirmed that the latter strains formed three genetically distinct groups. Each group was considered to represent a distinct species based on nucleotide sequences of the D1/D2 domains of the 26S rRNA gene and the internal transcribed spacer (ITS) region. Phylogenetic analyses of sequence data placed the putative novel species in a clade with R. acheniorum and the dimorphic smut fungus Farysia chardoniana. A novel anamorphic genus, Farysizyma, is created to accommodate the three undescribed species, which were named Farysizyma itapuensis, Farysizyma setubalensis and Farysizyma taiwaniana. A new combination, Farysizyma acheniorum, is proposed for R. acheniorum, which may represent the yeast-phase anamorph of Farysia thuemenii.
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