Molecular phylogeny and diversity of Fusarium endophytes isolated from tomato stems.

Plant tissues are a known habitat for two types of Fusarium species: plant pathogens and endophytes. Here, we investigated the molecular phylogeny and diversity of endophytic fusaria, because endophytes are not as well studied as pathogens. A total of 543 Fusarium isolates were obtained from the inside of tomato stems cultivated in soils mainly obtained from agricultural fields. We then determined partial nucleotide sequences of the translation elongation factor-1 alpha (EF-1α) genes of the isolates. Among the isolates from tomato, 24 EF-1α gene sequence types (EFST) were found: nine were classified as being from the Fusarium oxysporum species complex and its sister taxa (FOSC, 332 isolates), seven from the F. fujikuroi species complex (FFSC, 75 isolates) and eight from the F. solani species complex (FSSC, 136 isolates). To determine more characteristic details of the tomato isolates, we isolated 180 fusaria directly from soils and found 95% of them were nested within the FOSC (82 isolates; five EFSTs), FFSC (21 isolates; six FESTs) and FSSC (68 isolates; 11 EFSTs). These results suggested that the dominant Fusarium endophytes within tomato stems were members of the same three species complexes, which were also the dominant fusaria in the soils.

Improving the culturability of freshwater bacteria using FW70, a low-nutrient solid medium amended with sodium pyruvate.

Bacterial culture based on the use of plate media is an effective method for investigating bacterial populations in the environment. To improve the culturability of bacteria from freshwater lakes, we developed a new medium, FW70, which contains sodium pyruvate, casamino acids, and lake water and is solidified using gellan gum. To test the importance of these components, we prepared a series of media in which one or more of the components was absent. Water was sampled 31 times from 3 Japanese lakes and was passed through a membrane filter (pore size = 0.45 microm) to remove fast-growing microbes before the water was spread onto the plates. In most cases, significantly larger numbers of bacterial colonies were detected on FW70 than on other media. Furthermore, to test the practicality of FW70, we compared it with standard nutrient agar and R2A agar. In all cases, the culturability was significantly greater on FW70 than on standard nutrient agar or R2A agar. Some isolates recovered by means of FW70 belonged to bacteria that had not previously been classified. These results suggest that FW70 improves the culturability of freshwater bacteria and can be used for the isolation of novel bacteria as a result of the filtration step.

Fow2, a Zn(II)2Cys6-type transcription regulator, controls plant infection of the vascular wilt fungus Fusarium oxysporum.

The filamentous fungus Fusarium oxysporum is a soil-borne parasite that causes vascular wilts in a wide variety of crops by directly penetrating roots and colonizing the vascular tissue. In previous work, we generated the non-pathogenic mutant B137 of the melon wilt pathogen F. oxysporum f. sp. melonis by using restriction enzyme-mediated integration (REMI) mutagenesis. Molecular characterization of B137 revealed that this mutant has a single-copy plasmid insertion in a gene, designated FOW2, which encodes a putative transcription regulator belonging to the Zn(II)2Cys6 family. The REMI mutant B137 and other FOW2-targeted mutants completely lost pathogenicity, but were not impaired in vegetative growth and conidiation in cultures. Microscopic observation of infection behaviours of green fluorescent protein (GFP)-marked wild-type and mutant strains revealed that the mutants were defective in their abilities to invade roots and colonize plant tissues. FOW2 is conserved in F. oxysporum pathogens that infect different plants. The FOW2-targeted mutants of the tomato wilt pathogen F. oxysporum f. sp. lycopersici also lost pathogenicity. Nuclear localization of Fow2 was verified using strains expressing Fow2-GFP and GFP-Fow2 fusion proteins. These data strongly suggest that FOW2 encodes a transcription regulator controlling the plant infection capability of F. oxysporum pathogens.