Didactyl tracks of paravian theropods (Maniraptora) from the ?Middle Jurassic of Africa.

BACKGROUND: A new dinosaur tracksite from ?Middle Jurassic sediments of the Irhazer Group on the plains of Agadez (Rep. Niger, northwest Africa) revealed extraordinarily well preserved didactyl tracks of a digitigrade bipedal trackmaker. The distinct morphology of the pes imprints indicates a theropod trackmaker from a paravian maniraptoran closely related to birds. METHODOLOGY/PRINCIPAL FINDINGS: The early age and the morphological traits of the tracks allow for description of the new ichnotaxon Paravipus didactyloides. A total of 120 tracks are assigned to 5 individual trackways. The 'medium-sized' tracks with an average footprint length of 27.5 cm and footprint width of 23.1 cm are deeply imprinted into the track bearing sandstone. CONCLUSIONS/SIGNIFICANCE: A comparison with other didactyl tracks gives new insights into the foot morphology of advanced maniraptoran theropods and contributes to knowledge of their evolutionary history. The new ichnotaxon takes an important position in the ichnological fossil record of Gondwana and the mid-Jurassic biota worldwide, because it is among the earliest known records of paravian maniraptorans and of didactyl theropod tracks from Africa.

Calystegines in potatoes with genetically engineered carbohydrate metabolism.

Calystegines are hydroxylated nortropane alkaloids derived from the tropane alkaloid biosynthetic pathway. They are strong glycosidase inhibitors and occur in vegetables such as potatoes, tomatoes, and cabbage. Calystegine accumulation in root cultures was described to increase with carbohydrate availability. Whether this is indicative for the in planta situation is as yet unknown. Potatoes are model plants for the study of carbohydrate metabolism. Numerous transgenic potato lines with altered carbohydrate metabolism are available, but rarely were examined for alterations in secondary metabolism. In this study, calystegine accumulation and expression of biosynthetic enzymes were related to genetic modifications in carbohydrate metabolism in potato tubers. Tubers contained more soluble sugars due to overexpression of yeast invertase in the apoplast or in the cytosol, or due to antisense suppression of sucrose synthase. It is shown that the major part of calystegines in tubers originated from biosynthesis in plant roots. Yet, tuber calystegine levels responded to genetic alterations of carbohydrate metabolism in tubers. The strongest increase in calystegines was found in tubers with suppressed sucrose synthase activity. Transcripts and enzyme activities involved in calystegine biosynthesis largely concurred with product accumulation. Whole plant organs were examined similarly and displayed higher calystegines and corresponding enzyme activities in roots and stolons of plants with enhanced soluble sugars. Increases in calystegines appear to be linked to sucrose availability.

Immunolocalisation of two tropinone reductases in potato (Solanum tuberosum L.) root, stolon, and tuber sprouts.

Tropinone reductases (TRs) are essential enzymes in the tropane alkaloid biosynthesis, providing either tropine for hyoscyamine and scopolamine formation or providing pseudotropine for calystegines. Two cDNAs coding for TRs were isolated from potato (Solanum tuberosum L.) tuber sprouts and expressed in E. coli. One reductase formed pseudotropine, the other formed tropine and showed kinetic properties typical for tropine-forming tropinone reductases (TRI) involved in hyoscyamine formation. Hyoscyamine and tropine are not found in S. tuberosum plants. Potatoes contain calystegines as the only products of the tropane alkaloid pathway. Polyclonal antibodies raised against both enzymes were purified to exclude cross reactions and were used for Western-blot analysis and immunolocalisation. The TRI (EC 1.1.1.206) was detected in protein extracts of tuber tissues, but mostly in levels too low to be localised in individual cells. The function of this enzyme in potato that does not form hyoscyamine is not clear. The pseudotropine-forming tropinone reductase (EC 1.1.1.236) was detected in potato roots, stolons, and tuber sprouts. Cortex cells of root and stolon contained the protein; additional strong immuno-labelling was located in phloem parenchyma. In tuber spouts, however, the protein was detected in companion cells.

Overexpression of tropinone reductases alters alkaloid composition in Atropa belladonna root cultures.

The medicinally applied tropane alkaloids hyoscyamine and scopolamine are produced in Atropa belladonna L. and in a small number of other Solanaceae. Calystegines are nortropane alkaloids that derive from a branching point in the tropane alkaloid biosynthetic pathway. In A. belladonna root cultures, calystegine molar concentration is 2-fold higher than that of hyoscyamine and scopolamine. In this study, two tropinone reductases forming a branching point in the tropane alkaloid biosynthesis were overexpressed in A. belladonna. Root culture lines with strong overexpression of the transcripts contained more enzyme activity of the respective reductase and enhanced enzyme products, tropine or pseudotropine. High pseudotropine led to an increased accumulation of calystegines in the roots. Strong expression of the tropine-forming reductase was accompanied by 3-fold more hyoscyamine and 5-fold more scopolamine compared with control roots, and calystegine levels were decreased by 30-90% of control. In some of the transformed root cultures, an increase of total tropane alkaloids was observed. Thus, transformation with cDNA of tropinone reductases successfully altered the ratio of tropine-derived alkaloids versus pseudotropine-derived alkaloids.