Not the outside but the inside matters : functional analysis of Capricious and Tartan during Drosophila tracheal morphogenesis

The tubular structures, which transport essential gases, liquids, or cells from one site to another, are shared among various divergent organisms. These highly organized tubular networks include lung, kidney, vasculature and mammary gland in mammals as well as trachea and salivary gland in Drosophila melanogaster. Many questions regarding the tubular morphogenesis cannot be addressed sufficiently by investigating the mammalian organs because their structures are extremely complex and therefore, systematic analyses of genetic and cellular programs guiding the development is not possible. In contrast, the Drosophila tracheal development provides an excellent model system since many molecular markers and powerful tools for genetic manipulations are available. Two mechanisms were shown to be important for the outgrowth of tracheal cells: the FGF signaling pathway and the interaction between the tracheal cells and the surrounding mesodermal cells. The Drosophila FGF ligand encoded by branchless (bnl) is localized in groups of cells near tracheal metameres. The tracheal cells expressing the FGF receptor breathless (btl) respond to these sources of FGF ligand and extend towards them. However, this FGF signaling pathway is not sufficient for the formation of continuous dorsal trunk, the only muticellular tube in tracheal system. Recently, it was found out that single mesodermal cells called bridge-cells are essential for the formation of continuous dorsal trunk as they direct the outgrowth of dorsal trunk cells towards the correct targets. The results in this PhD thesis demonstrate that a cell adhesion molecule Capricious (Caps), which is specifically localized on the surface of bridge-cells, plays an essential role in guiding the outgrowing dorsal trunk cells towards their correct targets. When caps is lacking, some bridge-cells cannot stretch properly towards the adjacent posterior tracheal metameres and thus fail to interconnect the juxtaposing dorsal trunk cells. Consequently, discontinuous dorsal trunks containing interruptions at several positions are formed. On the other hand, when caps is ectopically expressed in the mesodermal cells through a twi-GAL4 driver, these mesodermal cells acquire a guidance function through ectopic caps and misguide the outgrowing dorsal trunk cells in abnormal directions. As a result, disconnected dorsal trunks are formed. These loss- and gain-of-function studies suggest that Caps presumably establishes the cell-to-cell contact between the bridge-cells and the tracheal cells and thereby mediates directly the guidance function of bridge-cells. The most similar protein known to Caps is another cell adhesion molecule called Tartan (Trn). Interestingly, trn is expressed in the mesodermal cells but not in the bridge-cells. When trn is lacking, the outgrowth of not only the dorsal trunks but also the lateral trunks are disrupted. However, in contrast to the ectopic expression of caps, the misexpression of trn does not affect tracheal development. Whereas Trn requires only its extracellular domain to mediate the matrix function, Caps requires both its extracellular and intracellular domains to function as a guidance molecule in the bridge-cells. These observations suggest that Trn functions differently from Caps during tracheal morphogenesis. Presumably, Trn mediates a matrix function of mesodermal cells, which support the tracheal cells to extend efficiently through the surrounding mesodermal tissue. In order to determine which domains dictate the functional specificity of Caps, two hybrid proteins CapsEdTrnId, which contains the Caps extracellular domain and the Trn intracellular domain, and TrnEdCapsId, which consists of the Trn extracellular domain and the Caps intracellular domain, were constructed. Gain of function and rescue experiments with these hybrid proteins suggest on one hand that the extracellular domains of Caps and Trn are functionally redundant and on the other hand that the intracellular domain dictates the functional specificity of Caps. In order to identify putative interactors of Caps, yeast two-hybrid screening was performed. An in vivo interaction assay in yeast suggests that Ras64B interacts specifically with the Caps intracellular domain. In addition, an in vitro binding assay reveals a direct interaction between an inactive form of Ras64B and the Caps intracellular domain. ras64B, which encodes a small GTPase, is expressed in the mesodermal cells concurrently as caps. Finally, a gain-of-function study with the constitutively active Ras64B suggests that Ras64B presumably functions downstream of Caps. All these results suggest consistently that the small GTPase Ras64B binds specifically to the Caps intracellular domain and may thereby mediate the guidance function of Caps.

Spechte als ökologische Indikatoren in Natur- und Wirtschaftswäldern im Bialowiera-Wald (Ostpolen)

Die vorliegende Untersuchung analysiert die Eignung der "Spechtgemeinschaft" als ökologische Indikatorengruppe und formuliert vor dem Hintergrund der Ergebnisse Forderungen und Empfehlungen für einen "spechtgerechten" Umgang mit Wäldern. Die Habitatnutzung von sieben Spechtarten beim Nahrungserwerb wurde über einen Zeitraum von zwei Jahren in urwaldartigen und forstlich genutzten Beständen verschiedener Waldgesellschaften systematisch beobachtet. Das Untersuchungsgebiet ist der Bialowieza-Wald im äußersten Osten Polens, wo in enger räumlicher Nachbarschaft Natur- und Wirtschaftswaldflächen bearbeitet werden konnten. Die Beobachtungen erfolgten zwischen Anfang März 1999 und Ende Februar 2001 und wurden zu allen Jahreszeiten durchgeführt. Vier der insgesamt sechs Probeflächen repräsentieren die wichtigste Laubwaldgesellschaft des Gebietes, das Tilio-Carpinetum, die übrigen zwei die wichtigste Nadelwaldgesellschaft, das Peucedano-Pinetum. Die Hälfte der zwischen 42 und 54 ha großen Probeflächen lag im streng geschützten Urwaldreservat des Bialowieza-Nationalparkes, die übrigen in forstlich genutzten Waldbeständen. Zusätzlich wurde ein 2,5 km langes Transekt durch bewirtschafteten Erlen-Eschen-Auenwald und sehr naturnahen Erlenbruch bearbeitet. Die Probeflächen wurden in ein Raster aus 50x50m großen Quadranten unterteilt. Zur Beobachtung der Spechte beim Nahrungserwerb erfolgten 21 Begehungen je Probefläche bzw. Transekt. Die Probeflächen wurden dazu auf parallelen Linien mit Abständen von je 100m begangen, Startpunkt und Startrichtung wurden variiert. Zur Charakterisierung der Vegetation und Bestandesstruktur erfolgten Erhebungen zur Baumartenzusammensetzung, Größenklassenverteilung der Bäume, Totholzanteil und Krautvegetation. 1332 Beobachtungen von Spechten beim Nahrungserwerb konnten ausgewertet werden. Der Buntspecht wurde in allen Flächen am häufigsten gesehen. Mittel-, Weißrücken- und Kleinspecht wurden überwiegend in den Tilio-Carpineten beobachtet, in den Naturwäldern häufiger als in den bewirtschafteten Beständen. Der Dreizehenspecht wurde im Nadelwald und stärker mit Fichten durchmischtem Laubwald angetroffen. Bei Schwarz- und Grauspecht konnte keine klare Vorliebe für bestimmte Waldgesellschaften ermittelt werden. Der Buntspecht ernährte sich vor allem im Herbst und Winter überwiegend von fetthaltigen Samen und wurde dann meist beim Bearbeiten von Fichten- oder Kiefernzapfen in Schmieden beobachtet. Der Mittelspecht suchte als "Sammelspecht" seine Nahrung vor allem an den Oberflächen der Stämme und Äste. Klein-, Weißrücken-, Dreizehen- und Schwarzspecht traten als Hackspechte in Erscheinung. Die wenigen Daten zum Grauspecht reichen nicht zur Ermittlung der bevorzugten Nahrungserwerbstechnik aus. Bei Bunt-, Mittel- und Weißrückenspecht konnte eine deutliche Vorliebe für die Stieleiche als Nahrungsbaum nachgewiesen werden. Der Dreizehenspecht ist jedoch die einzige der beobachteten Arten mit einer weitgehenden Spezialisierung auf eine bestimmte Baumart, er nutzte in allen Waldgesellschaften meist die Fichte. Insgesamt bevorzugten die Spechte Bäume mit großen Stammdurchmessern, beim Kleinspecht ist diese Vorliebe allerdings nur schwach ausgeprägt. Totholz wurde von Weißrücken-, Dreizehen- und Kleinspecht bei der Nahrungssuche bevorzugt, vom Mittelspecht jedoch nur gelegentlich genutzt. Beim Buntspecht zeigte der Totholz-Nutzungsanteil erhebliche Unterschiede zwischen verschiedenen Baumarten. Liegendes Totholz spielte in den Tilio-Carpineten im Vergleich zu stehendem Totholz und toten Teilen lebender Bäume nur eine geringe Rolle für Nahrung suchende Spechte.

Diversity and nutritional properties of Pakistani dates: implications for sustainable value chain and decent living perspectives of rural households

Date palm (Phoenix dactylifera L.) occupies almost three percent of the total worldwide cultivated area, with an annual production of seven million tonnes (t). Pakistan is an ideal place for the cultivation of date palm due to its sandy loam soil and semi-arid climate. In 2012, Pakistan produced 600,000 t of dates, on an area of 95,000 ha. Baluchistan province is the country’s top date producer, followed by Sindh, Punjab and Khyber Pakhtunkhwa (KPK) provinces. More than 300 date varieties are known to exist in Pakistan and some commercially important cultivars are: Karbalaen, Aseel, Muzawati, Fasli, Begum Jhangi, Hillawi, Dashtiari, Sabzo, Koharaba, Jaan Swore, Rabai and Dhakki. Six districts from the four provinces of Pakistan (Jhang, Muzaffargarh and Bahawalpur (Punjab), Dera Ismail Khan (KPK), Khairpur (Sindh) and Panjgur (Baluchistan)) with largest area under date palm cultivation were selected to conduct socio-economic surveys including the income sources of date palm growers. A structured questionnaire with open-ended and closed questions was used for face-to-face interviews of 170 date palm growers. At each location after selection of a first farmer through a local guide, the former was requested to provide names and addresses of three other date growers in his area. From these three names, one was randomly selected for the next sampling. Additionally, date palm fronds and fruits of all available cultivars were collected for morphological and nutritional analyses. Soil samples were collected from the groves for subsequent chemical and physical analyses. Almost all farmers used dates as a food item for their families and some were using low quality dates as a feed for their livestock. Apart from dates, other date palm components (trunk, spadix, frond, inflorescence and seed) were used by date palm growers as a raw material for making many by-products for their families. Date palm had a major contribution in the income of households, 24% received 91-100% of their income from date palms. More than half of the surveyed farmers had date palm groves, but scattered plantations, home gardens and intercropping systems with cereal and other fruits were also present. Dhakki, Muzawati, Aseel, and Karbalaen were the most important commercial cultivars grown in the provinces of KPK, Baluchistan, and Sindh. Aseel, Karoch, Haleni, Karbalaen, and Muzawati cultivars had the most firm fruit and good total soluble sugar, calcium and magnesium contents. The amount of magnesium found in dates of studied cultivars ranged from 0.143 to 0.876 mg g-1. A great variation in frond morphology was recorded among the cultivars. Fruit length and fruit weight was highest in Dhakki date, making it visually more attractive for customers in addition to its good nutritional properties. The seed weight of the studied cultivars ranged from 0.7-2.0 g, while Desi dates had largest seed, making them less attractive for marketing. However, in terms of nutritional value and fruit size, most of the investigated varieties can compete with globally important commercial dates.