No description available.

No description available.

The seedling-infecting pathogen Ustilago bullata Berk. is a naturally occurring biological control agent for cheatgrass (Bromus tectorum L.). The effects of temperature and nutrients on pathogen teliospore germination behavior and the effects of temperature on host seed germination were examined. The effects of temperature on sporidial proliferation, host infection in a temperature-controlled environment and in a field setting for eight populations were investigated. The infection success of Ustilago bullata on Bromus tectorum in cultivated fields as a function of seeding date, inoculation method, inoculum density, supplemental watering, and litter was also investigated. Teliospores germinated faster on potato dextrose agar than on water agar. Teliospores germinated slowly at temperatures far from the optimum of 15 and 20 C. There were among population variations in teliospore germination and sporidial proliferation, but differences among populations were much more pronounced at temperatures below 15 C. Infection also decreased and varied far from the optimum with almost no infection at 2.5 C in a controlled-environment and in the field for the December-planted seeds. Warmer early fall rather than the colder late fall was suitable for successful infection. This agreed with both laboratory and controlled-environment experiments. Intratetrad mating was observed with teliospores at 2.5 C. Teliospore germination tracked seed germination closely with teliospore germination rate exceeding the host seed germination rate over the range of 10 to 25 C where both were measured. Below 10 C, teliospore germination rate fell below host seed germination. This phenomenon was associated with lower infection percentages, suggesting that teliospore germination needed to be ahead of the seed for maximum infection. Inoculum density was positively correlated with infection rate. Litter significantly increased infection, while supplemental watering significantly increased plant establishment. Since teliospores from different populations showed similar germination patterns at temperatures typical of autumn seedbeds in the Intermountain West, it may not be necessary to use locally-adapted pathogen populations in biological control program. A biocontrol program is most likely to be effective under a scenario where autumn precipitation permits emergence of most of the host seed bank as a fall cohort.

Cytoplasmic dynein is a microtubule-dependent motor protein which participates in numerous cellular processes. The motor complex consists of two heavy chains, intermediate, light intermediate and 3 families of light chains. Dynein is able to bind to these accessory chains as well as to regulatory proteins which enables the motor protein to fulfil such a variety of cellular processes. The associated light chains participate in long-distance organelle and vesicle transport in interphase and in chromosome segregation during mitosis. However, how these light chains control the activity of the motor protein is still unknown. In this study, I combine molecular genetics and live cell imaging to elucidate the role of the associated dynein light intermediate and light chains in dynein behaviour and early endosome (EE) motility in hyphal interphase cells as well as the anchorage of dynein to the microtubule (MT) plus-end in interphase and mitotic cells. I show that the dynein light intermediate chain (DLIC) as well as the light chain 2 (DLC2, Roadblock) are involved in dynein processivity and EE movement in interphase. The downregulation of either protein results in short hyphal growth which could be caused by a decreased runlength of EE and dynein. In addition, both proteins participate in dynein anchorage to the microtubule plus-end in interphase and mitosis as well as in spindle elongation during mitosis. Each protein causes a decrease of the motor protein dynein at MT plus-ends. Surprisingly, I found only minor or no defects in LC8 or Tctex mutants in the observed functions of dynein. LC8 seems to affect the dynein but not the EE runlength. In this case, dynein is still able to move into the bipolar MT array from where kinesin3 is able to take over EEs and move them towards the cell center. In contrast, Tctex has no effect on dynein or EE runlength or any other observed dynein function in hyphal cells. However, it causes a reduction in spindle elongation. Taken together, DLIC and DLC2 are important for dynein behaviour in long distance transport as well as in spindle positioning and elongation during mitosis. Furthermore, I studied the involvement of the dynein regulators Lis1 and NudE as well as the plus-end binding protein Clip1 (Clip-170 homologue) in the anchorage of dynein to the astral microtubule plus-ends during mitosis. The disruption of the anchorage complex at the astral MT plus-end causes a decrease in dynein number at this site and therefore slower spindle elongation in Anaphase B. Taken together, all three proteins are involved in anchorage of dynein to the astral microtubule tip and the subsequent spindle elongation. Furthermore, these findings also show that Ustilago maydis evolved two different mechanisms to anchor the motor protein to MT plus-ends in hyphal and mitotic cells. The plus-end binding protein Peb1 (EB1 homologue) and the dynein regulator dynactin mediate the dynein anchorage in hyphal cells whereas in mitotic cells the plus-ends binding protein Clip1 and the dynein regulators Lis1 and NudE anchor dynein to astral MT plus-ends.