This paper summarizes work done in this laboratory over the last two years on the cloning of microsporidian rRNA by homology PCR and its subsequent use in diagnostic tests and phylogenetic studies. Using highly conserved primers in the 16S or small subunit rRNA (SSU-rRNA) these genes were cloned from human intestinal biopsies with transmission electron microscopy proven Enterocytozoon bieneusi and Septata intestinalis. The SSU-rRNA genes were then used to design and test several primer pairs for the diagnosis of microsporidian infection. Utilizing the polymerase chain reaction and primers V1 and EB45Ü Ent. bieneusi infected duodenal aspirates or intestinal biopsies could be detected. Using V I and SI500 infection with S. intestinalis could be detected. In addition to diagnostic tests, phylogenetic relationships were examined using sequence data from the fragment amplified by PCR by primer 530f in the SSU-rRNA and primer 580r in the large subunit rRNA. This data supported the placement of S. intestinalis in the family Encephalitozoonidae. In addition, it confirmed that Encephalitozoon cuniculi, E. hellem and S. intestinalis are distinct organisms. These techniques have broad applications to the study of other microsporidia and the development of a molecular phylogeny.
Incorporation of labelled CO2, 3-phosphoglycerate (PGA), phosphoenolpyruvate (PEP) and pyruvate into hexane extractable rabber ffactions in the cut shoots of guayule {Parthenium argentatum Gray) was determined in order to evaluate the role of photosynthesis in providing precursors for rubber biosynthesis. DCMU inhibited the incorporation of labelled CO2 and PGA into rubber. The incorporation of i'*C02 into rubber depended on irradiance. Enzymatic activities of phosphoglyceromutase, enolase, pyruvate kinase and pyruvate dehydrogenase complex found in purified chloroplasts from the leaves indicated the chloroplast autonomy for intraplastid acetyl coenzyme A formation. The enzymes related to the biosynthesis of isopentenyl pyrophosphate (IPP) were associated with both leaf and stem extracts. Rubber producing enzyme activities, námely IPP isomerase and rubber transferase, were abundantly localized in roots and stems of guayule while the leaves exhibited low activities of these enzymes. Hence the leaves of guayule play a major role in providing precursors for rubber formation in stems and roots.
In the temperate haptophyceaen Pavlova luíheri (Droop) Green and Pavlova sp. (not yet estimated) the dominant pigments were chlorophyll (Chl) a (57 %), fucoxanthin (17 %) and diadinoxanthin (13 %); other estimated pigments were p-carotene, Chl c, cA-fucoxanthin and diatoxanthin. UV-A of 1.7 W m-2 had no effect on pigmentation of P. luíheri, but led to a reduction of the pigment contents of Pavlova sp. The damaging effect of a high UV-A dose (11.0 W m'^) was more pronounced than that of UV-B irradiance. The strongest reduction of pigment concentrations of both Pavlova species was observed after an exposure to UV-A (11.0 W m'^) plus UV-B (1.2 W m-2). Under this combined irradiation an increase of the diatoxanthin content and a decrease of the diadinoxanthin content was found.
The review deals with the comprehensive analysis of ultraviolet-B (UV-B) induced alterations in photosynthetic processes of higher plants and possible protection mechanisms. Between two photosystems, photosystem 2 (PS2) is main target for UV- B radiation stress. Depending on the mode of treatment, the oxidising side of PS2 and reaction centres are mainly affected by UV-B treatment. As response to UV-B stress plants develop some inherent adaptive mechanisms which decide the extent of the overall damage.