Molekularbiologische Funktionsanalysen von Thioredoxinen aus Dictyostelium discoideum und Identifizierung neuer Interaktionspartner durch das Two-Hybrid-System

Thioredoxins are small, regulatory proteins with a mass of approximately 12 kDa and a characteristic conserved active center, which is represented in the pentapeptide trp-cys-gly-pro-cys. Up to now it is not possible to present a complete list of thioredoxin interaction partners because there is no predictable sequence in the target enzymes where thioredoxins can interact with. To get closer information about the functions and possible interaction partners of the three thioredoxins from the social soil amoeba Dictyostelium discoideum (DdTrx1 - 3) we have chosen two different strategies. In the first one the thioredoxin levels in the cell should changed by different mutants. But both the antisense technique as well as the creation of knock out mutants were not appropiate strategies in this case. Just an thioredoxin overexpressing mutant results in a developmental phenotype which allows some conclusions for possible functions of the thioredoxin in Dictyostelium discoideum. The second strategie was the two hybrid system where thioredoxin interactions partners can identified systematically. After a screening with a cDNA library from Dictyostelium 13 potential interaction partners could be detected, among them a ribonucleotid reductase, TRFA, two different cytochrome c oxidase subunits, filopodin, three ribosomal proteins, the elongationfactor 1a and the alcohol dehydrogenase from yeast. The verification of the interaction between thioredoxin and these two hybrid clones happened indirectly by a dobble mutant of thioredoxin 1, where the cysteines in the active center were replaced by redox-inactive serins. Further examinations of two choosen candidates resulted that the alcohol dehydrogenase from yeast is a thioredoxin-modululated enzym and that there is an interaction between the elongationfactor 1a and the thioredoxin 1 from Dictyostelium discoideum.

Effects of time-controlled and continuous grazing on total herbage mass and ground cover

Grazing practices in rangelands are increasingly recognized as a management tool for environmental protection in addition to livestock production. Long term continuous grazing has been largely documented to reduce pasture productivity and decline the protective layer of soil surface affecting environmental protection. Time-controlled rotational grazing (TC grazing) as an alternative to continuous grazing is considered to reduce such negative effects and provides pasture with a higher amount of vegetation securing food for animals and conserving environment. To research on how the grazing system affects herbage and above ground organic materials compared with continuous grazing, the study was conducted in a sub-tropical region of Australia from 2001 to 2006. The overall results showed that herbage mass under TC grazing increased to 140% in 2006 compared with the first records taken in 2001. The outcomes were even higher (150%) when the soil is deeper and the slope is gentle. In line with the results of herbage mass, ground cover under TC grazing achieved significant higher percentages than continuous grazing in all the years of the study. Ground cover under TC grazing increased from 54% in 2003 to 73%, 82%, and 89% in 2004, 2005, and 2006, respectively, despite the fact that after the high yielding year of 2004 herbage mass declined to around 2.5 ton ha^(−1) in 2005 and 2006. Under continuous grazing however there was no significant increase over time comparable to TC grazing neither in herbage mass nor in ground cover. The successful outcome is largely attributed to the flexible nature of the management in which grazing frequency, durations and the rest periods were efficiently controlled. Such flexibility of animal presence on pastures could result in higher water retention and soil moisture condition promoting above ground organic material.