Perspectives on genetic resources at the Fungal Genetics Stock Center.

The Fungal Genetics Stock Center has been banking and distributing resources for work with genetically characterized fungi since 1960. While most of the collection consists of strains of Neurospora, an NIH model filamentous fungus, the past fifteen years has seen the collection expand to include plant and human pathogenic fungi. The use of the resources in the collection has grown over the last 10 years as well, reflecting both a growth in research using standardized materials as well as the development of new materials through molecular genetic technology. This growth is not limited to newly deposited materials, however, and includes renewed interest in particular classes of strains with characteristics that were not recognized when they were originally deposited. One significant example is the use of strains carrying the osmotic-2 lesion in Neurospora crassa. This, and other utilization trends, underscores the need to provide strong support to the continued and expanded biobanking effort in the US

Fungal Genetics Stock Center Catalogue of Strains

Catalogue of Strains, 11th edition, 2006, supplement to Fungal Genetics Newsletter No. 53. This catalogue contains lists of materials held by the Fungal Genetics Stock Center

Fungal Genetics Stock Center Catalogue of Strains, 10th Edition

Catalogue of Strains, 10th edition, 2004, supplement to Fungal Genetics Newsletter No. 51. This catalogue contains lists of materials held by the Fungal Genetics Stock Center

Enhancing the seed germination process of Montezuma cypress (Taxodium mucronatum Ten.)

Montezuma cypress (Taxodium. mucronatum) is an ecological, cultural and economically valuable riparian tree species. Two experiments evaluating the effectiveness of various seed treatments were conducted to identify germination best practices and to evaluate the dynamics of the germination process. Seeds were collected on two occasions, one year apart, from the only remaining natural T. mucronatum tree stand in the United States. The seeds were subjected to various soaking and stratification conditions. Across all treatments, germinability ranged between approximately 30%-40%, with slightly higher values occurring among the second seed cohort. Overall, no significant differences in germinability were detected in either study, however, soaking seeds in water for 96 hours and stratifying them in moist conditions for 3 weeks significantly accelerated the germination process. Seeds soaked briefly in a NaOH solution followed by a 48-hour water soak demonstrated more synchronous germination than other treatments. Control conditions in which seeds were not soaked or stratified exhibited the slowest germination. These findings are consistent with previous evidence showing that T. mucronatum seeds do not exhibit physiological dormancy and that treatments promoting seed water imbibition enhance the germination process. This study adds to the limited available research on T. mucronatum propagation practices and offers novel data on the germination parameters of seeds sourced from a natural U.S. stand, rather than seeds from few scattered individual trees, as in previous reports. Seed germination recommendations garnered from this study can improve nursery production of T. mucronatum to enhance ecological restoration efforts and ornamental production

Work in the dark to harvest large liquid-grown cultures

Biochemical purification of low-abundance proteins from Neurospora crassa often requires collection of \u3e100 g wet weight of mycelial mass. For purification of the dynein motor from N. crassa, 4 to 8 one liter liquid cultures are inoculated with 1 x 106 conidia/ml at 3:00 pm and incubate overnight at 28°C with shaking. At 9:00 am the next morning, mycelia (10- 15 g/flask) are collected by filtration using a new cellulose filter for each flask (Fisherbrand P8). Unfortunately, we frequently find that mycelia are easily collected from the first one to three flasks, however, mycelia cannot be harvested from the remaining flasks because the filters become clogged. We have determined that this is a light-dependent phenomenon. If the incubators are covered in black trash bags for the overnight incubation and the lab lights are not turned on during the morning harvesting period, we no longer see any clogging of filters. We suspect that light-induction of hydrophobins is the cause of the clogging of cellulose filters (Lauter et al. 1992)

Fantastic growth as the FGSC turns 50

Comparative Medicine - OneHealth and Comparative Medicine Poster SessionFounded in 1960, the Fungal Genetics Stock Center enters it's fiftieth year of operation during a period of tremendous growth. The collection has more than doubled since moving to UMKC in 2004 and has added new materials that reach out beyond it's traditional constituency. Among these are deletion sets for Neurospora, Cryptococcus and Candida as well as molecular genetic tools for working with industrial fungi, model organisms, and plant and human pathogens. With distribution growing every year, the FGSC sends materials to scientists in over 35 countries every year; approximately half of our orders are from within the US. In addition to being part of an NIH funded multi-institution Functional Genomics Program for Neurospora, the FGSC is involved in cutting edge genomics research with collaborators at the US DOE Joint Genome Institute. The FGSC and its staff are actively involved in national and international societies and ad hoc working groups fostering the development of collection resources in the US and around the world

Demonstration that the Neurospora crassa mutation un-4 is a single nucleotide change in the tim16 gene encoding a subunit of the mitochondrial inner membrane translocase

The Neurospora crassa temperature sensitive mutation known as un-4 has been shown by a map-based complementation approach to be a single nucleotide change in the open reading frame of the mitochondrial inner membrane translocase subunit tim16 (NCU05515)

Kinesin and dynein mutants provide novel insights into the roles of vesicle traffic during cell morphogenesis in Neurospora

AbstractBackground: Kinesin and cytoplasmic dynein are force-generating molecules that move in opposite directions along microtubules. They have been implicated in the directed transport of a wide variety of cellular organelles, but it is unclear whether they have overlapping or largely independent functions.Results: We analyzed organelle transport in kinesin and dynein single mutants, and in a kinesin and dynein double mutant of Neurospora crassa. Remarkably, the simultaneous mutation of kinesin and dynein was not lethal and resulted in an additive phenotype that combined the features of the single mutants. The mutation of kinesin and dynein had opposite effects on the apical and retrograde transport, respectively, of vesicular organelles. In the kinesin mutant, apical movement of submicroscopic, secretory vesicles to the Spitzenkörper – an organelle in the hyphal apex – was defective, whereas the predominantly retrograde movement of microscopic organelles was only slightly reduced. In contrast, the dynein mutant still had a prominent Spitzenkörper, demonstrating that apical transport was intact, but retrograde transport was essentially inhibited completely. A major defect in vacuole formation and dynamics was also evident. In agreement with the observations on apical transport, protein secretion into the medium was markedly inhibited in the kinesin mutant but not in the dynein mutant.Conclusions: Transport of secretory vesicles is necessary but not sufficient for normal apical extension. A component of retrograde transport, presumably precursors of the vacuole system, is also essential. Our findings provide new information on the role microtubule motors play in cell morphogenesis and suggest that kinesin and cytoplasmic dynein have largely independent functions within separate pathways

Identification of the Neurospora crassa mutation un-10 as a point mutation in a gene encoding eukaryotic translation initiation factor 3, subunit B.

The Neurospora crassa temperature-sensitive mutant known as un-10 has been shown by a map-based complementation approach to be a single nucleotide change in the open reading frame of the eukaryotic translation initiation factor 3b (NCU02208.3)