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  • Rural Digital Europe
  • 2013-2022
  • Open Access
  • Doctoral thesis
  • Rural Digital Europe

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  • image/svg+xml art designer at PLoS, modified by Wikipedia users Nina, Beao, JakobVoss, and AnonMoos Open Access logo, converted into svg, designed by PLoS. This version with transparent background. http://commons.wikimedia.org/wiki/File:Open_Access_logo_PLoS_white.svg art designer at PLoS, modified by Wikipedia users Nina, Beao, JakobVoss, and AnonMoos http://www.plos.org/
    Authors: Imran, Muhammad;

    Seed reserves are the primary source of mineral nutrients for early seedling development. ?Nutrient seed priming? is a technique in which seeds are soaked in nutrient solution and subsequently dried back to initial moisture content for storage. It is an efficient approach to increase seed nutrient contents along with priming effects to improve seed quality, germination speed and seedling establishment. Various abiotic stresses, such as sub-optimal temperature, drought, submergence and soil pH extremes can seriously affect seedling establishment and nutrient acquisition at early growth stages. This thesis focused on the role and contribution of nutrient seed priming in plant growth and nutritional status in maize and soybean under conditions of limited nutrient availability and low root-zone temperature. Protocols for nutrient seed priming with Zn, Mn, Zn+Mn, B and P were optimised for application in maize and soybean seeds (B and P priming in maize only). Optimum priming durations of 24 h (maize) and 12 h (soybean) were identified for both plant species but in instead of submerging seeds in priming solutions slow imbibition between filter papers was essential for soybean to minimise development of abnormal seedlings to avoid imbibition damage. Nutrient concentrations were calculated according to water uptake to double the natural seed reserves of the respective micronutrients and 50% increase in phosphorus. However, final uptake of the micronutrients was generally much higher (+500-1000%) while it was lower for P (+20%). In case of soybean this could be attributed to a high Zn and Mn binding capacity of the seed coat, which adsorbed up to 60% of the primed nutrients. Particularly, Zn and Zn+Mn priming stimulated plant growth in hydroponic culture systems and to a lower extent also on a soil with low availability of P, Zn and Fe. This was associated with a high shoot translocation of the primed nutrients (Zn and Mn), which was most expressed in the hydroponic culture system. Combined priming treatments with Zn and Mn were usually less effective than Zn priming alone, suggesting an antagonistic interaction. By contrast, mobility of primed B was extremely low and B priming was completely ineffective. In soil culture also P priming moderately increased shoot biomass production by 10-20 %. However the efficiency of P priming was largely limited by the high P demand of the plants. Low root zone temperature (RZT) at early spring is a limiting factor for maize production in Central and Northern Europe. Nutrient acquisition, nutrient uptake and particularly root growth are severely affected at low RZT and the consequences of these growth depressions are often not completely compensated until final harvest. Model experiments in hydroponics and soil culture revealed that maize nutrient seed priming with Zn, Mn and Fe is a promising strategy to diminish the deficiency of specific nutrients, such as Zn, Mn and also P and to maintain plant growth under low RZT stress. This was mainly attributed to significantly increased root growth and particularly fine root production in plants grown from nutrient-primed seeds. Improved net photosynthesis of primed plants was mainly related with increased leaf area and preliminary results suggest a higher tolerance to oxidative damage due to increased production of protective phenolics. Two independent field experiments under conditions of suboptimal temperatures during germination and early growth revealed an increase in grain yield of 10 ? 15 % for plants derived from Zn+Mn and Fe primed seeds. This finding demonstrates long-lasting persistence of priming effects. The molecular and physiological mechanisms behind require further investigation. Samenreserven sind die primäre Quelle für die Mineralstoffversorgung während der frühen Keimlingsentwicklung. Nährstoffpriming ist ein methodischer Ansatz, bei dem Saatgut in Mineralstofflösungen vorgequollen und anschließend zur weiteren Lagerung auf Ausgangsfeuchte zurückgetrocknet wird. Hierdurch ist es möglich, neben vorquellungsbedingten Primingeffekten auch die Mineralstoffreserven zur Verbesserung der Saatgutqualität zu erhöhen sowie die Auflauf-Geschwindigkeit und die Keimlingsentwicklung zu verbessern, die durch verschiedenste Stressfaktoren, wie suboptimale Temperaturbedingungen, Trockenheit, Staunässe und pH-Extreme beeinträchtigt werden kann. Die vorliegende Arbeit beschäftigt sich mit der Wirkung von Nährstoffpriming auf das Wachstum und den Ernährungsstatus von Mais und Soja unter Bedingungen eingeschränkter Nährstoffverfügbarkeit und niedriger Wurzelraumtemperatur. Für die Saatgutapplikation von Zink (Zn), Mangan (Mn), Mn+Zn, Bor (B), Eisen (Fe) und Phosphat (P) bei Mais und Soja wurden Primingprotokolle entwickelt. Bei beiden Pflanzenarten ergab sich eine optimale Behandlungsdauer von 24 h. Jedoch war es notwendig, das Soja-Saatgut anstelle von submerser Inkubation in den Nährlösungen, langsam in Nährlösungsgetränktem Filterpapier einzuquellen, um die Entwicklung abnormaler Keimlinge aufgrund von Quellungsschäden zu vermeiden. Anhand der Wasseraufnahme der Keinlinge wurden die Nährlösungskonzentrationen so berechnet, das sich nach Einquellen eine Verdopplung der natürlichen Mineralstoffreserven im Samen ergeben sollte. Jedoch wurden für die Mikronährstoffe erheblich höhere Aufnahmeraten gemessen (+500 - 1000%), während die Aufnahme von P geringer war (+ 20%). Bei Soja konnte gezeigt werden, das die erhöhten Mikronährstoffgehalte auf eine hohe Bindungskapazität der Zwei unabhängige Feldversuche bei suboptimaler Temperatur während der Keimphase und des Jugendwachstums ergaben Kornertragssteigerungen von 10-15 % bei Mais nach Saatgutpriming mit Zn+Mn oder Fe. Diese Ergebnisse belegen die Langzeitwirkung der Primingeffekte. Die molekularen und physiologischen Grundlagen erfordern allerdings noch weitere Untersuchungen.

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    Authors: Kirschner, Stefanie;

    Industrielle Röntgendurchleuchtungsgeräte lassen sich für die Untersuchung verschiedenster Fragestellungen anwenden. Das Spektrum der Anwendbarkeit reicht von der hochauflösenden in vivo-Bildgebung von z. B. Glioblastomen, zerebralen Hirngefäßen, abdominellen und thorakalen Organen in der Maus, bis hin zur hoch-auflösenden Untersuchung von Objekten wie z. B. Stents, Zähnen, Insekten oder auch einzelner Organe ex vivo. Desweitern zeichnet das hier eingesetzte industrielle Röntgengerät die Möglichkeit zur Strahlentherapie von Tieren aus, was mit herkömmlichen speziellen Kleintier-Computertomographen nicht möglich ist. Ziel dieser Dissertation war zum einen die Etablierung eines Glioblastom-Maus-Modells und anschließend die Überprüfung der Anwendbarkeit eines Mikro-CT für die wiederholte bildgebende Darstellung der Hirntumoren vor und nach Strahlentherapie. Darüber hinaus wurde die Mikro-CT für die Evaluierung sowohl der Verschlussrate von Aneurysmen als auch der artefaktfreien Darstellung der verwendeten Materialien im Modellversuch getestet. Wir konnten in beiden von uns durchgeführten Studien die Mikro-CT erfolgreich zur Bildgebung anwenden. Zur Etablierung des Glioblastom-Maus-Modells wurde die gesamte OP-Methode zur Tumorzellimplantation in das Gehirn zunächst an toten Mäusen durchgeführt. Anstatt Zellen wurde eine äquivalente Menge an mit Kontrastmittel vermengter Methylcellulose des später geplanten Injektionsvolumens an Zellen in das Gehirn appliziert. Anschließend wurde zur Erfolgskontrolle eine Röntgenaufnahme im Mikro-CT angefertigt. Anhand der Aufnahmen konnten wir die genaue Lokalisation und Ausdehnung der applizierten Menge beurteilen und unsere OP-Methode gegebenenfalls optimieren. Erst nach verlässlicher Reproduzierbarkeit der OP wurden humane Zellen in vivo in die rechte Hemisphäre von insgesamt 27 NOD/SCID/yc-/- Mäusen injiziert. Zwei Wochen nach Xenotransplantation erfolgte eine erste kontrasmittelgestützte Untersuchung im Mikro-CT zur Tumordetektion. Bei negativem Befund wurde bei den Tieren wöchentlich eine weitere Mikro-CT durchgeführt. Tiere mit positivem Befund wurden je nach geplantem Versuch nicht therapiert (n=21) oder mittels Strahlentherapie (n=6) behandelt. Alle Tiere wurden im weiteren Verlauf wöchentlich bis zum Versuchsende im Mikro-CT untersucht. Insgesamt wurden 103 kontrastmittelgestützte Untersuchungen im Mikro-CT durchgeführt. Der maximale Tumordurchmesser und das maximale Tumorvolumen wurden anhand der einzelnen CT-Datensätze bestimmt. Zusätzlich wurden die Gehirne histologisch aufgearbeitet um das maximale Volumen und den maximalen Durchmesser der Tumore anhand der Histologie zu berechnen. Die histologischen Ergebnisse wurden mit der jeweils letzten Mikro-CT (durchgeführt am Tag des Versuchsendes) verglichen. Die hervorragende Korrelation von maximalem Tumordurchmesser und Tumorvolumen ermittelt in der Mikro-CT mit den Ergebnissen aus der Histologie (Tumordurchmesser: 0,929, p < 0,001; Tumorvolumen: 0969, p < 0,001, n = 17) belegt die hohe Genauigkeit und damit Anwendbarkeit der Mikro-CT für ein Hirntumormodell. Die hohe Sensitivität (0,95) und Spezifität (0,71) belegen zusätzlich die Exaktheit der Mikro-CT zur Erkennung von Gehirntumoren in sehr kleinen Tieren wie Mäusen (PPV=0,91, NPV=0,83). Die zweite im Rahmen dieser kumulativen Dissertation eingereichte Arbeit befasst sich mit der Evaluation einer neuen Methode zur Embolisation von Aneurysmen. Hierzu wurden Aneurysmen mit einem dicht gewobenem Stent (sog. Flow diverter) überdeckt. Über einen zuvor im Aneurysma platzierten Mikrokatheter wurde das Aneurysma mit Mikropartikeln embolisiert, welche größer als die Maschen des flussmodulierenden Stents sind. Die Mikro-CT konnten wir erfolgreich einsetzen, um mit einer hohen Auflösung die 700–900 µm kleinen Partikel im Aneurysma darzustellen und die komplette Ausfüllung des Aneurysmas zu verifizieren. Auch die Maschendichte des Stents im Verhältnis zum Aneurysma konnte mittels Mikro-CT analysiert werden. Die Möglichkeit der wiederholten Darstellbarkeit von Gehirntumoren im zeitlichen Verlauf mittels Mikro-CT bietet eine Grundlage für weitere Versuchsvorhaben, wie z. B. die Visualisierung morphologischer Veränderungen des Tumors unter verschiedenen Therapieansätzen. Die erfolgreiche Evaluation der Verschlussrate von Aneurysmen mittels einer neuen Technik in der Mikro-CT ist ein wesentlicher Schritt, bevor es zu einer ersten Anwendung im lebenden Tier kommt. Beide hier vorgestellten Arbeiten tragen durch den erfolgreichen Einsatz eines bildgebenden Verfahrens erheblich zur langfristigen Senkung der Versuchstierzahlen bei. Zusammenfassend konnten wir durch die erfolgreiche Anwendung der Mikro-CT in den hier vorgestellten Arbeiten das Spektrum der Anwendbarkeit eines industriellen Mikro-CTs in der präklinischen Forschung deutlich erweitern. Micro-CT systems are being used with increasing frequency in preclinical research. The range of examinations includes high-resolution in vivo imaging of glioblastoma, cerebral vessels, abdominal and thoracic organs in mice as well as investigation of other small objects like stents, teeth, insects or single organs ex vivo or in vitro. In addition, micro-CT may be used for radiation therapy of small rodents like mice and rats, as shown in our studies. Aim of this thesis was to establish an orthotopic glioblastoma mouse model and to evaluate the applicability of a micro-CT for repetetive imaging of brain tumors in mice. We furthermore used a micro-CT to evaluate the distribution of microspheres used to treat a silicone aneurysm model. The glioblastoma mouse model was trained in dead mice first. Instead of tumor cells we first injected methylcellulose mixed with contrast agent to train intracerebral injections. The use of contrast agent allowed us to verify our injection technique by subsequent micro-CT. After successfully establishing the technique, human brain tumor cells were in injected into the right hemisphere of n = 27 NOD/SCID/yc-/- mice. Two weeks after xenotransplantation mice underwent first contrast-enhanced micro-CT. Animals with negative results in micro-CT received additional micro-CT scans weekly until brain tumors were detected. Animals were left untreated (control group), or were treated with radiation- or chemotherapy. During treatment all animals received weekly micro-CT scans until animals had to be sacrificed. The maximum diameter and volume of intracerebrally growing xenografts were measured. Brains of sacrificed mice were histologically worked up and maximum tumor diameter and tumor volume were measured again. Results from histology were compared to micro-CT data. The excellent correlation between maximum tumor diameter and tumor volume measured in micro-CT and histology (tumor diameter: 0.929, p < 0.001; tumor volume: 0.969, p < 0.001, n=17). Sensitivity and specificity of micro-CT were 0.95 and 0.71, respectively (PPV=0.91, NPV=0.83). This proved the high accurary of micro-CT for brain tumor detection in animals as small as mice. The second paper referenced to within this doctoral thesis describes the evaluation of the distribution of microspheres within a silicon aneurysm and the degree of aneurysm occlusion. For this purpose a silicon aneurysm was treated with a densely woven stent (a so called flow diverter). Additionally, the aneurysm was embolized with microspheres larger than the pores of the stent via a microcatheter previously placed in the aneurysm. Using our micro-CT we were able to visualize the 700-900 µm small microspheres, which allowed us to verify complete embolization of the aneurysm. Furthermore, the mesh density of the flow diverter in relation to the size of the microspheres was analyzed using the micro-CT. From our findings we conclude, that micro-CT is a versatile tool that may be used for imaging of a vast range of objects and thus propose that micro-CT will be used with increasing frequency in preclinical research.

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    Authors: Krishnan, Parvathy;

    Pathogenic fungi impose a great threat to the global food production. They are globally distributed and display a huge amount of phenotypic diversity. They demonstrate high adaptability to diverse environments and to fluctuating conditions within their hosts. However, not much is known about the mechanisms underlying these processes of adaptation. Deciphering the genetic basis of phenotypic diversity may help us to get a deeper insight into the mechanisms that regulate complex adaptive traits. Furthermore, these findings may help in improving strategies for an effective and sustainable management of pathogens. In my PhD study, I aimed to understand the genetic basis of adaptation to variable environments using Zymoseptoria tritici. Z. tritici causes septoria leaf blotch of wheat and is considered to be the most devastating fungal wheat pathogen in Europe. In the first chapter of my thesis, I studied the role of melanin in adaptation of Z. tritici. I, demonstrated that melanin is essential for protection against harmful chemical fungicides such as bixafen. Remarkably, I observed high phenotypic variability in melanin accumulation levels in different Zymoseptoria tritici strains, most probably reflecting its role in adaptation to variable environments. Using genetic mapping, I was able to demonstrate that variability in melanization levels between two Swiss strains of Z. tritici was mediated by differential expression of the transcription factor Zmr1 (Zymoseptoria melanin regulation 1). I identified nucleotide mutations in the promoter region and an insertion of transposable elements (TEs) upstream of the promoter region as two mechanisms responsible for this observed differential expression of Zmr1. My findings clearly demonstrated the significance of gene expression regulation, mediated by the insertion of TEs and SNPs, in optimizing the growth and adaptation of the fungus under variable environments. Furthermore, these adaptive changes are responsible for the phenotypic diversity that we observe in Z. tritici strains from all over the world. In the second part of my thesis, I analyzed the expression of several secreted peptidases of Z. tritici at distinct stages during the infection process of wheat plants. Using computational genetic methods, I also analyzed nucleotide data sets of the peptidase genes for evolutionary signatures of genetic adaptation. By combining both approaches, I aimed at obtaining a more detailed picture of the relevance and the role of each enzyme during the infection process. I was able to show that the peptidases belonging to MEROPS families A1 and G1 were significantly up regulated during the asymptomatic phase of infection and displayed signatures of accelerated evolution, suggesting their key role in suppression of plant defenses and host specialization. In the third part of my thesis, I evaluated the role of a cellulase-encoding gene (JGI Prot. ID: 76589) using molecular biology techniques. This cellulase was only expressed during the final necrotrophic phase of Z. tritici infection and the nucleotide sequences of the gene showed signatures of diversifying selection. This pattern is consistent with the hypothesis of evasion from host recognition during the first phases of infection and subsequent induction of host-cell death by the secretion of an array of different cellulase isoforms by the fungus. To test this hypothesis, I constitutively expressed the cellulase-encoding gene in Z. tritici during all infection stages by inserting a constitutive promoter (originally from the Glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans) upstream of the start codon of the cellulase gene. I was able to show that the progress of infection in this mutant was delayed compared to the non-modified Z. tritici isolates. This suggests that the cellulase acts as an elicitor molecule that is detected by the plant and triggers the defense machinery that will hinder the growth of the pathogen. In summary, my PhD work demonstrated the significance of fine-tuning of gene expression regulation of various genes in plant pathogenic fungi to facilitate adaptation to different environmental conditions and host colonization. Remarkably, I was able to show that gene expression fine-tuning resulted in large amounts of phenotypic diversity in adaptive traits.

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    ETH Zürich Research Collection
    Doctoral thesis . 2018
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      ETH Zürich Research Collection
      Doctoral thesis . 2018
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    Authors: Šumrada, Tanja;

    Kmetijstvo sodi med najpomembnejše dejavnike zmanjševanja biotske pestrosti v EU, vendar integracija varstva narave v Skupno kmetijsko politiko ostaja nezadostna. Namen doktorske disertacije je proučiti načrtovanje, učinke in vrednotenje slovenske kmetijske politike na področju ohranjanja narave. Analiza z modeli BRT je pokazala negativno povezavo med diverziteto ptic kmetijske krajine in neposrednimi plačili, okoljski ukrepi pa so imeli v obdobju 2008–2019 na diverziteto ptic šibek relativni vpliv. Diverziteta ptic je bila najvišja v odprti krajini na območjih Natura 2000 z visoko pestrostjo kmetijskih rastlin in nizko obtežbo z živino. Programsko-teoretična analiza slovenske kmetijske politike je pokazala številne vrzeli v opredelitvi intervencijske logike. Manjka predvsem opredelitev ciljev in kazalnikov, ki bi omogočali presojo uspešnosti ukrepanja. Integracija varstva narave je na strateškem nivoju omejena predvsem na prostovoljne ukrepe, strategije za preprečevanje konfliktov med naravovarstvenimi in drugimi cilji kmetijske politike pa večinoma niso opredeljene. Ocenili smo, da je bilo v programskem obdobju 2014–2020 varstvu narave namenjenih okrog 5 % (17,8–19,4 mio EUR) letnega proračuna slovenske kmetijske politike. Disertacijo zaokrožuje analiza možnosti za izboljšanje rezultatske naravnanosti kmetijsko-okoljskih ukrepov, ki je potekala s poskusom diskretne izbire (521 kmetov na območju Haloz in Krasa) ter kvalitativno tematsko analizo intervjujev s kmeti, raziskovalci, odločevalci in kmetijskimi svetovalci. Kmetje so izkazali statistično značilno večje preference do rezultatskih shem kot do shem s predpisanimi praksami ter naklonjenost do individualnega pristopa k svetovanju. Za uspešno uveljavitev rezultatskih shem je nujno izboljšanje podatkovnih podlag, raziskav trajnostnih proizvodnih modelov ter usposobljenosti kadrov v nadzornih in svetovalnih službah. Agriculture is one of the most important factors in reducing biodiversity in the EU. However, the integration of nature conservation into the Common Agricultural Policy remains inadequate. The purpose of the doctoral dissertation is to study the planning, effects and evaluation of the Slovenian agricultural policy in the field of nature conservation. An analysis using BRT models showed a negative link between the diversity of farmland birds and direct payments, while environmental measures had a weak relative influence on bird diversity in the period 2008–2019. Bird diversity was highest in open landscapes on Natura 2000 sites that had a high diversity of agricultural plants and low stocking density. A programme theory analysis of the Slovenian agricultural policy showed numerous gaps in the definition of the intervention logic. Particularly lacking is a definition of objectives and indicators that would enable an assessment of the effectiveness of interventions. At the strategic level, the integration of nature conservation is limited mainly to voluntary measures. Strategies for preventing conflicts between nature conservation and other objectives of agricultural policy are mostly not defined. We estimated that in the 2014–2020 programming period, approximately 5 % (17.8–19.4 million EUR) of the annual budget of the Slovenian agricultural policy was allocated to nature conservation. The dissertation is concluded with an analysis of the possibilities for improving the result-orientation of agri-environmental measures, which was conducted with a discrete choice experiment (521 farmers in the Haloze and Karst area) and a qualitative thematic analysis of interviews with farmers, researchers, decision-makers and agricultural advisers. The farmers showed statistically significant higher preferences for result-based schemes than for schemes with prescribed practices and a preference for an individual approach to knowledge transfer. In order to successfully implement result-based schemes, it is necessary to improve data bases, research into sustainable production models and the training of staff in supervisory and advisory services.

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    Authors: Wapelhorst, Britta;

    In der vorliegenden Doktorarbeit sollte a) die Expression sowie b) die zelluläre Lokalisation des SLC10A6 (SOAT) im humanen Hoden geklärt werden. Der SOAT zeigt im Vergleich mit weiteren Gewebetypen vergleichsweise hohe Expression im Hoden und weist eine Transportaffinität für sulfatierte Steroide auf. Sulfatierte Steroide können aufgrund ihrer hydrophilen Eigenschaften Zellmembranen nicht durch Diffusion passieren, sondern benötigen einen Transporter. Zudem sind sulfatierte Steroide biologisch nicht aktiv und können erst nach Desulfatierung an ihre entsprechenden Rezeptoren binden und eine biologische Antwort hervorrufen. Aufgrund seiner Eigenschaften ist der SOAT ein potentieller Kandidat für den Transport sulfatierter Steroide im humanen Hoden und somit für die lokale Bereitstellung von Androgenen und Östrogenen. Die Untersuchungen wurden an Hodenbiopsien von Patienten durchgeführt, die eine normale Spermatogenese aufwiesen und Hodenbiopsien von Patienten mit gestörter Spermatogenese, bis hin zu einer völligen Abwesenheit von Keimzellen, dem Sertoli Cell Only Syndrom. Der Nachweis der Expression des SOAT erfolgte mittels TaqMan®-RT-qPCR, RT-PCR nach Laser-assistierter Mikrodissektion und In-situ- Hybridisierung. In den Untersuchungen der vorliegenden Arbeit konnte durch TaqMan®-RT-qPCR und RT-PCR nach Laser-assistierter Mikrodissektion gezeigt werden, dass der SOAT in Hodenbiopsien mit normaler Spermatogenese exprimiert wird. Der zelluläre Nachweis des SOAT erfolgte mittels In-situ-Hybridisierung in Keimzellen, den primären Spermatozyten. Für den Nachweis des SOAT Proteins wurden im Rahmen dieser Dissertation sechzehn Antikörper eingesetzt. Eine spezifische Färbung konnte jedoch bei keinem Antikörper beobachtet werden. Untersuchungen von Fietz et al. (2013) konnten das SOAT-Protein schließlich mittels eines Antikörpers gegen das Soat-Protein der Maus Soat(m329-344) in Spermatozyten und teilweise in Spermatiden nachweisen. Da es sulfatierten Steroiden nicht möglich ist, die Blut-Hoden-Schranke ohne die Hilfe eines Transporters zu überwinden und so die Keimzellen zu erreichen, stellt sich die Frage nach der Funktion des SOAT in den Keimzellen des humanen Hodens. Der Nachweis der Expression des SOAT erfolgte auch in drei Hodenbiopsien mit Spermatogenestörungen, die mit einer Infertilität einhergehen. Aufgrund der signifikant geringeren Expression des SOAT in Hodenbiopsien mit einer Hypospermatogenese kann vermutet werden, dass eine signifikant geringere Expression des SOAT mit einem verringertem Transport von sulfatierten Steroiden und in Folge mit einer reduzierten Bereitstellung von lokal verfügbaren Androgenen und Östrogenen einhergeht (Fietz et al. 2013). Diese Fragestellung könnte durch Untersuchungen der Spermatogenese von Slc10a6-knockout Mäusen weiterführend untersucht werden. It was the aim of this doctoral thesis to elucidate a) the expression and b) the cellular localization of the sodium-dependent organic anion transporter SLC10A6 (SOAT) in human testis. SOAT is predomiantly expressed in human testis and shows specific transport for sulfated steroids. Because of their hydrophilic nature, sulfated steroids are not able to pass cell membranes and therefore need the help of a transporter. Moreover, sulfated steroids are not biologically active, only after desulfatation they can bind to their receptors to induce a biological response. SOAT is a potential candidate not only for transport of sulfated steroids in the human testis, but also for a local supply of androgens and estrogens. The study included testicular biopsis showing normal spermatogenesis and spermatogenic impairment, including Sertoli cell only syndrome. Detection of SOAT mRNA was performed by TaqMan®-RT-qPCR, RT-PCR after laser-assisted microdissection and in situ hybridization. TaqMan®-RT-qPCR and RT-PCR after LAM detected expression of SOAT in testicular biopsies showing normal spermatogenesis. SOAT was localized in germ cells, in primary spermatocytes by in situ hybridization. For verification of SOAT protein distribution 16 antibodies were tested but none could give answer to the question of protein localization. Fietz et al. (2013) employed a new antibody against SOAT protein of mice (Soatm329-433) and detected SOAT in spermatocytes and spermatid of various stages. Due to unabillity for sulfated steroids to pass the blood-testis-barrier, the function of SOAT in spermatocytes is still not known. SOAT could be verified in three testicular biopsies showing impaired spermatogenesis resulting in infertility. Fietz et al. confirmed a significant lower expression of SOAT in testicular biopsies showing hypospermatogenesis. It is suggested that the reduction or lack of SOAT expression might be related to a decreased local supply with androgens and estrogens (Fietz et al. 2013). Further studies with Slc10a6-knockout mice could corroborate this hypothesis.

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    Authors: Kumar, Sandhya Sasi;

    Since its founding as a social welfare state, India has been meandering through “a life of contradictions” when it comes to social and economic equality. A potent indicator of this reality is the state of food and nutrition security across the country. India’s meager progress comes on the heels of rapid economic growth over the last two decades and a slew of public programs have attempted to address the multi-faceted nature of food and nutrition security. These efforts have included subsidized grains for households through the Targeted Public Distribution System (TPDS), village health, nutrition and education programming through the Integrated Child Development Services (ICDS), and Nutrition Rehabilitation Centers (NRCs) to treat cases of severe acute malnutrition (SAM). Though these programs have been scaled across the country, numerous governance challenges and institutional failures have persisted. The latest development in India’s food and nutrition policy landscape is the National Food Security Act (hereafter referred to as the Act) of 2013, which leverages existing national programs, including the TPDS and ICDS, to grant legal entitlements to nearly 70% of the total population. This Act was the fruit of a rights-based movement in India that demanded for over a decade to codify social and economic rights. However, the question arises as to whether this new Act been able to achieve its goal to ensure food and nutrition security by strengthening the governance of these programs? This is the central question of the present thesis. This thesis investigates how the Act has reformed the implementation of food and nutrition programs in the states of Jharkhand, Madhya Pradesh and the capital of New Delhi. The four key objectives of this study are to investigate 1) what changes are prescribed by the Act; 2) how effective these reforms have been in addressing persistent governance challenges; 3) what challenges remain and why; and 4) what strategies could be used to address these gaps. A qualitative case study approach was applied, which involved the following data collection methods: key informant and in-depth interviews, focus group discussions, participant observation, matrix ranking and process net-mapping, an innovative participatory tool that maps complex institutional processes and actors. This thesis is comprised of nine chapters. The first chapter introduces the puzzle of India’s nutritional status and explores why government efforts to tackle this problem have had tepid results. The second chapter presents the conceptual framework that defines governance challenges and an institutional framework of demand and supply side factors that contribute to effective policy and programming. Chapters three and four present an overview of the TPDS, ICDS and NRCs, and review the existing literature on the governance issues of these programs. Chapter 5 lays out the study design, methods and ethical protocols. Chapters six through eight explore each of the three programs and present key findings across the selected study sites. The last chapter presents cross-cutting conclusions for these programs in the wake of the Act and proposes a way forward to address institutional gaps in tackling food and nutrition security. Seit der Gründung des Wohlfahrtsstaates erlebt Indien eine Reihe von Widersprüchen, was die soziale und wirtschaftliche Gerechtigkeit des Landes angeht. Ein aussagekräftiger Indikator dieser Realität ist der Stand der Ernährungsicherheit im Land. Indiens dürftiger Fortschritt in der Ernährungssicherung folgt einer zwei Jahrzehnte andauernden Periode rasantem Wirtschaftswachstum, sowie einer Reihe von öffentlichen Programmen, die versuchen, die vielschichtigen Aspekte von Ernährung zu adressieren. Zu diesen Bemühungen gehören subventioniertes Getreide für Haushalte durch das gezielte öffentliche Vergabesystem TPDS, dörfliche Gesundheits-, Ernährungs- und Bildungsprogramme durch die Integrierten Dienstleistungen zur Entwicklung von Kindern ICDS und Rehabilitierungszentren für Fälle von schwerer akuter Mangelernährung (NRCs). Obwohl diese Programme auf ganz Indien ausgeweitet wurden, bestehen weiterhin institutionelle Mängel sowie zahlreiche Governance-Herausforderungen. Die jüngste Entwicklung in der Indischen Ernährungspolitik ist das Gesetz zur nationalen Ernährungssicherheit NFSA aus dem Jahr 2013. Das NFSA nutzt zwei bestehende nationale Programme, TPDS und ICDS, um fast 70% der Bevölkerung Rechtsansprüche zu gewähren. Dieses Gesetz war das Ergebnis einer größeren rechtebasierten Bewegung in Indien, die seit über einem Jahrzehnt das Ziel hat, soziale und wirtschaftliche Rechte zu kodifizieren. Es stellt sich die Frage, ob dieses neue Gesetz es ermöglicht hat, durch eine Stärkung der Governance der bestehenden Programme einen Lebenszyklusansatz in der Ernährungssicherung zu etablieren. Diese Doktorarbeit verwendet einen Ansatz vergleichender Fallstudien, um zu untersuchen, wie das NFSA die Umsetzung der Ernährungsprogramme in den Staaten Jharkhand, Madhya Pradesh und der Hauptstadt Neu Delhi reformiert hat. Die vier Hauptziele der Dissertation sind: 1) die Änderungen, die durch das NFSA vorgeschrieben wurden, zu beschreiben 2) die Effektivität dieser Reformen bei der Bewältigung von Governance-Herausforderungen zu erklären, 3) die weiterhin bestehenden Governanceprobleme und Gründe dafür zu erklären, und 4) Strategien zu erarbeiten, die zur Behebung dieser Defizite beitragen können. Diese vier Aspekte wurden untersucht mittels Befragung von Schlüsselpersonen, Tiefeninterviews, Fokusgruppendiskussionen, teilnehmender Beobachtung, Matrix-Ranking und Prozessnetzkartierung (process net-mapping), ein innovatives und partizipatives Instrument, das komplexe institutionelle Prozesse und Akteure abbildet. Die vorliegende Doktorarbeit besteht aus neun Kapiteln. Das erste Kapitel gibt eine Einleitung in die Rätselhaftigkeit der Ernährungslage in Indien, und warum staatliche Bemühungen dieses Problem anzugehen, nur dürftige Ergebnisse erzielt haben. Das zweite Kapitel stellt einen konzeptionellen Rahmen bezüglich Governance-Herausforderungen und einen institutionellen Rahmen bezüglich Nachfrage- und Angebotsfaktoren, die zu wirkungsvollen Politikmaßnahmen und Programmgestaltung beitragen, dar. Kapitel drei und vier geben einen Überblick über TPDS, ICDS und NRCs, und über die bestehende Literatur bezüglich der Governance-Herausforderungen dieser drei Programme. Kapitel fünf beschreibt das Studiendesign, die angewandten Methoden und die Forschungsprotokolle. Kapitel sechs bis acht untersuchen jeweils eins der drei Programme und erklären Schlüsselergebnisse über die drei ausgewählten Untersuchungsstandorte hinweg. Die Dissertation schließt mit einer übergreifenden Zusammenfassung für diese Programme vor dem Hintergrund des neuen Gesetzes und gibt wegweisende Vorschläge wie man diese institutionellen Defizite zur Verbesserung der Ernährungssicherheit adressiert.

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    Authors: Yilmaz, Gülenay;

    The fact that rapid diagnosing of especially seed-borne pathogens and their control is the first step to obtain healthy crop. The aims of the study were to optimize the analyzing methods in detecting presence of Plasmopara halstedii (Farl.) Berl. & De Toni requiring analysis at the time of importation in the sunflower seed using current molecular methods that were much more advantageous than that of classical methods, and to evaluate potential variations in sensitivities of the pathogen to metalaxyl and mefenoxam. Calculations related to classical PCR (agarose gel electrophoresis) were optimized by referring to Appendix 2 of the EPPO Standard Protocols while those related to real-time PCR (the hydrolyze probes) were optimized by referring to Appendix 3 of the EPPO Standard Protocols. Moreover, SyberGreen Mastermix Kit protocol was used for optimization for the study of real-time PCR (SyberGreen) that is not present in the EPPO Standard Protocols. Thus, it was provided that all three molecular protocols were confirmed for better applicable and practical ways for the quarantine laboratories in our country. Furthermore, the first molecular studies on direct detection of the pathogen from sunflower seeds through these optimization was conducted, as well. Reduced sensitivities of P. halstedii to metalaxyl are well-known throughout the world. In this study, it was showed that the similar reduced sensitivities of the pathogen had been determined for mefenoxam that is an isomer of metalaxyl. There were found no differences statistically and both fungicides were placed in the same group because of resulting from cross-resistance between. Özellikle, tohum kaynaklı olan ve tohumla taşınabilen etmenlerin hızlı tanılanması ve bunların kontrol altına alınması sağlıklı bir ürün elde etmenin ilk aşamasını oluşturmaktadır. Bu tez çalışmasının amacı, ithalat aşamasında analiz şartı olan tohum kaynaklı Plasmopara halstedii (Farl.) Berl. & De Toni)'nin ayçiçeği tohumundaki varlığının klasik yöntemlere göre çok daha avantajlı moleküler yöntemler kullanılarak optimizasyonunun gerçekleştirilmesi ve metalaxyl ile mefenoxam arasındaki duyarlılıklarının değerlendirilmesidir. Klasik PCR (agaroz jel elektroforez) ile ilgili hesaplamaları EPPO Standart Protokolleri Appendix 2 ve Real Time PCR (hidroliz probları) ile ilgili hesaplamalar ise EPPO Standart Protokolleri Appendix 3 referans alınarak optimize edilmiştir. Ayrıca, EPPO Standart Protokollerinde yer almayan Real time PCR (SyberGreen) çalışmasında ise, SyberGreen Mastermix kit protokolü esas alınarak optimize edilmiştir. Böylece, ülkemiz karantina laboratuarları içerisinde her üç moleküler protokülün daha uygulanabilir olması sağlanmıştır. Ayrıca, bu optimizasyon ile etmenin ayçiçeği tohumundan direkt tespitine yönelik ilk moleküler çalışma da gerçekleştirilmiştir. P. halstedii izolatlarının metalaxyl'e duyarlılıklarının azaldığı tüm dünyada bilinmektedir. Bu çalışmada da, söz konusu patojenin metalaxyl'in izomeri olan mefenoxam'a karşı da duyarlılıklarının azaldığı belirlenmiş ve aralarında çapraz dayanıklılık olduğu için metalaxyl ile aynı grupta yer almıştır.

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    Authors: Hailu, Teweldemedhn Gebretinsae;

    Ethiopia is a major beekeeping country located in northeast Africa where several evolutionary lineages of Apis mellifera contact. A unique practice of honey bee colony marketing which involves broad agro-ecological zones (AEZs) is a developing trend in the northern part of the country such as Tigray region in association with apicultural development. Several studies based on classical morphometry on the Ethiopian honey bee subspecies classification debated from the unique Apis mellifera simensis to five others. Moreover, the genetic diversity, adaptation, gene flow and inter-relationships of the honey bees between AEZs were not disentangled – a challenge for planning sustainable apicultural development and conservation. Therefore, this study was conducted to elucidate the honey bees of Ethiopia in a context of apicultural transformation using integrated methods: morphometrics, genetics, colony market survey and metadata analyses on beekeeping development. The results of geometric morphometric analyses confirmed that Ethiopian honey bees represented by Apis mellifera simensis references belong to a separate lineage (Y) compared to A, O, M and C, and the present sample belonged to Y. This supported the hypothesis of five major honey bee lineages of the honey bee Apis mellifera. Similarly, a maximum likelihood phylogenetic tree analysis based on the mitochondrial COI-COII showed that most of the Ethiopian honey bees belong to lineage Y. However, a substantial proportion of the samples from the northern part of the country clustered with lineage O, which support the hypothesis that there is close contact between Y and O. Both geometric morphometry and classical morphometry differentiated the Ethiopian honey bees from all references including A. m. monticola, A. m. scutellata, A. m. jementica, A. m. adansonii but grouped with A. m. simensis. Genetically, five DraI haplotypes (COI-COII) were found to be randomly distributed across AEZs, indicating a substantial gene flow. Consequently, the level of genetic differentiation among the Ethiopian honey bee subpopulations defined by local areas and AEZs was generally low based on r7-frag nuclear marker, which is identified to be associated with adaptation to habitat elevation in East African honey bees. Similarly, nucleotide diversity consistently decreased with increasing elevation – indicating a reduced effective population size in the highlands. Results obtained from colony market survey showed that the honey bee swarms are reproduced in a few highlands and re-distributed throughout the region. Colony buyers have preferences of color and AEZ of origin of the honey bees, which led to a one-way flow and eroded the overall level of genetic differentiation. However, a marked differentiation was detected between the highland and lowland honey bees in relic communities where an allelic length polymorphism was observed as a signature of local adaptation. Altogether, Ethiopian honey bees belong to the lineage Y and subspecies A. m. simensis, and are characterized by a high level of gene flow enhanced by colony marketing; but a conserved signature of local adaptation to higher elevations was identified in less disturbed communities. Further studies based on genome-wide analyses and field experiments, focusing on undisturbed communities, can provide more insights into adaptation, admixture and management implications. Sustainable bee breeding and extension services that enable local beekeeping without colony trade and transportation will help to promote apiculture and genetic conservation. Äthiopien ist ein Land mit vielfältigen Ökosystemen im Nordosten Afrikas, in denen viele evolutionäre Linien von Apis mellifera zusammenkommen. In der Region Tigray (im Norden von Äthiopien) werden Honigbienenvölker auf zentralen Märkten gehandelt. Mehrere Studien zur Klassifikation der Unterarten von äthiopischen Honigbienen auf der Grundlage der klassischen Morphometrie stellten unterschiedliche Hypothesen über die Anzahl dieser Unterarten auf. Darüber hinaus war der Mangel an Informationen über genetische Vielfalt, Anpassung und Genfluss eine Herausforderung für die Planung einer nachhaltigen Entwicklung der Imkerei. Ziel dieser Doktorarbeit war es, diese Honigbienen und den Handel mit ihnen auf der Grundlage von Morphometrie, Genetik und Marktuntersuchungsanalysen in einen Zusammenhang zu stellen. Die Ergebnisse bestätigten, dass äthiopische Honigbienen, die durch Apis mellifera simensis-Referenzen repräsentiert werden, von den Linien A, O, M und C morphologisch unterschieden werden können und zu der Linie Y gehören, was die Hypothese von fünf großen Honigbienen-Linien unterstützt. In ähnlicher Weise zeigte die mitochondriale COI-COII-Analyse, dass die meisten äthiopischen Honigbienen der Linie Y angehören. Ein erheblicher Anteil der Proben aus dem nördlichen Teil des Landes gruppierte sich jedoch mit der Linie O, was die Hypothese eines engen Kontaktes zwischen den Linien Y und O stützte. Hinsichtlich der Unterarten unterschieden die geometrische Morphometrie und die klassische Morphometrie die äthiopischen Honigbienen von allen Referenzen einschließlich A. m. monticola, A. m. scutellata, A. m. jementica, A. m. adansonii, aber gruppierten mit A. m. simensis. Genetisch wurden fünf DraI-Haplotypen identifiziert, die zufällig über die agro-ecological zones (AEZs) verteilt waren, was auf einen erheblichen Genfluss hinweist. Folglich war die genetische Differenzierung der äthiopischen Proben insgesamt gering. Die Abnahme der Nukleotiddiversität in den Subpopulationen von A. mellifera in höher gelegenen Habitaten deutet auf eine reduzierte effektive Populationsgröße im Hochland hin. Die Markterhebung ergab, dass sich die Honigbienenvölker in einigen Hochländern vermehren und durch die Vermarktung in der gesamten Region von Tigray verteilt werden. Eine Ursache hierfür ist, dass die Käufer die Farbe und die AEZ der Herkunft bevorzugen. Dies führte zu einem unidirektionalen Fluss von genetischem Material und verringert die genetische Differenzierung. In lokalen, ungestörten Gebieten konnte jedoch eine deutliche Differenzierung zwischen der Hochland- und der Tieflandhonigbiene festgestellt werden. Dabei wurde ein Allellängenpolymorphismus als Zeichen der lokalen Anpassung beobachtet. Zusammenfassend lässt sich feststellen, dass äthiopische Honigbienen der Linie Y und der Unterart A. m. simensis zugehörig sind. Ein hoher Genfluss zwischen Subpopulationen wird durch Koloniemarketing verstärkt; aber eine konservierte Signatur der lokalen Anpassung an höhere Lagen wurde in ungestörten Gebieten identifiziert. Weitere Forschungen auf der Grundlage genomweiter Analysen und Feldexperimente, die sich auf die ungestörten lokalen, Gebiete konzentrieren, können weitere Einblicke in Anpassung, Durchmischung und Management liefern. Nachhaltige Bienenzucht und Beratungsdienste, die eine lokale Imkerei ohne Völkerhandel ermöglichen, werden zur Förderung der Imkerei und der genetischen Erhaltung beitragen.

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    Authors: van Iersel, W.K.;

    Natural lowland rivers are dynamic environments with a high ecological value. However, 90% of the European and North-American river floodplains are in a degraded state. The functions of floodplains are strongly determined by land cover and they often compete for space in narrowed floodplains. Integrated river management (IRM) tries to take care of floodplains in such way that land cover is optimized for multiple functions. For IRM, monitoring is essential to capture the dynamics, to evaluate changes, and to document the state of floodplains over time. The main objective of this thesis was to establish remote-sensing methods for the monitoring of floodplain land cover over multiple spatial and temporal scales. Several remote-sensing based solutions have been developed for the monitoring of land-cover dynamics in river floodplains and tested in floodplains of the lower Rhine. The phenological change of floodplain vegetation over the course of one year was studied using temporal profiles of its height and greenness. Using multitemporal UAV images, vegetation height was determined with an accuracy similar to much more expensive airborne LiDAR data. Multitemporal elevation models yielded meaningful profiles of greenness and vegetation height over time, which enabled discriminating the different land-cover types. The same dataset combined with a powerful machine learning model (Random Forest) yielded unprecedented high classification accuracies for floodplain vegetation (> 90%), even for similar vegetation types such as grassland and herbaceous vegetation. This method is a practical and highly accurate solution for monitoring areas of a few square kilometres. For large-scale monitoring of floodplains, the same method is recommended, but with data from airborne platforms covering larger extents. Land-cover change over the course of five years was studied for a 100-km river section using satellite images. Using an object-based approach, a sequential deviation of a land-cover object from its class mean was used to detect land-cover change. For most classes the method was unsuccessful (accuracy 75%. The developed method has important advantages, such as high observation frequency, independence of repeated land-cover classification, and fast processing. At sub-daily frequency, it was assessed how accurate water temperature in a floodplain side channel can be documented from thermal UAV maps. The associated habitat suitability for native and alien fish assemblages was estimated based on the produced temperature maps. Water surface temperatures were mapped four times during a hot summer day with an overall RMSE of 0.53 oC. During the day, temperatures in the side channel increased rapidly to values detrimental for many fish species. The study showed that thermal imagery from UAVs is an efficient and accurate information source to monitor spatiotemporal patterns of thermal habitat suitability. The presently available range of spaceborne and airborne platforms and sensors offers great opportunities to collect information on land-cover change across a range of spatial and temporal scales. This may advance our management of floodplains and help us recovering and protecting these rich ecosystems and the benefits they provide us.

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    Authors: Strigens, Alexander Carl Georg;

    Since the introduction of maize into Europe, open-pollinated varieties of flint maize were cultivated across the continent. Natural selection promoted adaptation to the climatic conditions prevailing in the different regions. With the advent of hybrid breeding in Europe during the 1950’s, some of the genes responsible for the specific adaptations of the landraces to abiotic and biotic stress were captured in the first developed inbred lines, but most of their genetic diversity is still untapped. Development of inbred lines out of this material by recurrent selfing is very tedious due to strong inbreeding depression. In contrast, the doubled-haploid (DH) technology allows producing fully homozygous lines out of landraces in only one step. This allows their precise characterization in replicated trials and identification of new genes by genome wide association (GWA) mapping. In this study we genotyped a set of 132 DH lines derived from European Flint landraces and 364 elite European flint (EU-F), European dent (EU-D) and North-American dent (NA-D) inbred lines with 56,110 single nucleotide polymorphism (SNP) markers. The lines were evaluated in field trials for morphologic and agronomic traits and GWA mapping was performed to identify underlying quantitative trait loci (QTL). In particular, our objectives were to (1) develop a robust method for quantifying early growth with a non-destructive remote-sensing platform, (2) evaluate the importance of early growth performance of inbred lines with regard to their testcross performance, (3) determine the potential of GWA mapping to identify genes underlying early growth and cold tolerance related traits, (4) evaluate the phenotypic and genotypic diversity recovered in the DH lines derived from the landraces, (5) estimate the effect of the DH method on the recovered genetic diversity, (6) identify new genes by GWA mapping in the DH lines derived from landraces, and (8) discuss the potential of DH lines derived from landraces to improve the genetic diversity and performance of elite maize germplasm. A phenotyping platform using spectral reflectance and light curtains was used to perform repeated measurements of biomass and estimate relative growth rates (RGR) of the DH and inbred lines, as well as of two testcrosses of 300 dent inbred lines. The DH lines derived from the landraces Schindelmeiser and Gelber Badischer had the highest RGR followed by EU-F lines, DH lines derived from Bugard, EU-D lines and, finally, NA-D lines. For inbred lines, whole plant dry matter yield (DMY) was positively correlated with RGR (r = 0.49), whereas this relation was weaker in the testcrosses (r = 0.29). RGR of the inbred lines correlated with RGR of their testcrosses (r = 0.42), but it had no influence on testcross DMY. A set of 375 EU-F, EU-D and NA-D lines were further evaluated in growth chambers under chilling (16/13°C) and optimal (27/25°C) temperatures. Photosynthetic and early growth performance were estimated for each treatment and an adaptation index (AI) built as the chilling to optimal performance ratio. Nineteen QTL were identified by GWA mapping for trait performance and AI. Candidate genes involved in ethylene signaling, brassinolide, and lignin biosynthesis were found in their vicinity. Several QTL for photosynthetic performance co-located with previously reported QTL and the QTL identified for shoot dry wieght under optimal conditions co-located with a QTL for RGR. Comparison of the DH lines derived from landraces with the EU-F lines showed that genotypic variances in single DH populations were greater than in the EU-F breeding population. A high average genetic distance among the DH lines derived from the same landrace as well as a rapid decay of linkage disequilibrium suggests a high effective population size of the landraces. Because no systematic phenotypic differences were observed between the landraces and synthetic landraces obtained by intermating the corresponding DH lines, the expected purge of lethal recessive alleles during the DH production did neither improve grain yield performance nor affect the recovered genetic diversity. Performing GWA in the DH lines derived from landraces as well as the EU-F, and EU-D lines allowed the identification of 49 QTL for 27 traits. A larger set of DH lines derived from more landraces might solve problems arising from population structure and allow a much higher power for the detection of new alleles. In conclusion, the introgression of DH lines derived from landraces into the elite breeding material would strongly broaden its genetic base. However, grain yield performance was 22% higher in EU-F lines than in the DH lines derived from landraces. Selection of the best DH lines would allow partially bridging this yield gap and marker-assisted selection may allow introgression of positive QTL without introducing negative features by linkage drag. Seit der Einfuhr von Mais aus der „neuen“ Welt nach Europa, wurden offen abblühende Flint-Mais Populationen auf dem gesamten Kontinent angebaut. Durch natürliche Selektion passten sich diese Landsorten an die Klimate des Kontinents an. In den Anfängen der Hybridzüchtung wurden Gene und Allele, die für diese spezifische Anpassung an biotische und abiotische Stressfaktoren verantwortlich sind, in den ersten Inzuchtlinien nur teilweise fixiert. Der Grossteil der genetischen Vielfalt der Landsorten blieb jedoch ungenutzt, da die Entwicklung von Inzuchtlinien aus diesem Material wegen besonders starker Inzuchtdepression sehr mühsam ist. Demgegenüber erlaubt es die seit etwa 10 Jahre eingesetzte Methode der Erzeugung von Doppel-Haploiden (DH), vollständig homozygote Linien aus Landsorten in einem einzigen Schritt zu entwickeln. Diese DH-Linien können in wiederholten Feldversuchen sehr präzise evaluiert werden. Dies vereinfacht die Kartierung von Genen mithilfe der Genom-weiten Assoziations-Kartierung (GWA) enorm. In der vorliegenden Studie wurden 132 DH-Linien aus europäischen Landsorten, 364 Inzucht-linien aus Nordamerikanischem Dent (NA-D), europäischem Flint (EU-F) und europäischem Dent (EU-D) Zuchtmaterial mit 56110 genetischen Markern genotypisiert. Agronomische Eigenschaften der DH-Linien und Elite-Inzuchtlinien wurden in Feldversuchen evaluiert und mittels GWA kartiert, um vorteilhafte Gene zu identifizieren. Zu unseren Zielen gehörten insbesondere (1) die Entwicklung einer robusten, nicht-destruktiven Methode zur Erfassung der Jugendentwicklung mittels Sensoren, (2) die Untersuchung des Zusammenhangs zwischen der Jugendentwicklung der Linien per se und deren Testkreuzungen, (3) die Erforschung von GWA zur Identifikation von Kühletoleranz- und Jugendentwicklungs-Genen in Elite-Inzuchtlinien, (4) die Evaluierung der aus den Landsorten mittels der DH-Methode geborgene phänotypische und genetische Vielfalt, (5) die Abschätzung eines möglichen Einfluss der DH-Methode auf der genetischen Vielfalt der DH-Linien, (6) die Entdeckung neuer Gene in den DH-Linien aus Landsorten mittels GWA, und (7) die Ermittlung des Potentials von DH-Linien aus Landsorten, um die Leistung und genetische Diversität des modernen Zuchtmaterials zu verbessern. Die Biomasse und relative Wachstumsrate (RGR) der DH-Linien und Elite-Inzuchtlinien sowie je zwei Testkreuzungen von 300 Dent Inzuchtlinien wurden mit Lichtschranken und spektraler Reflektion geschätzt. Die DH-Linien aus den Landsorten Schindelmeiser und Gelber Badischer wiesen die höchste RGR auf, gefolgt von EU-F Linien, DH-Linien aus Bugard, EU-D Linien und zuletzt NA-D Linien. Die Gesamttrockenmasse der Linien war mit deren RGR positiv korreliert (r = 0.49), während diese Korrelationen für die Testkreuzungen schwächer ausfiel (r = 0.29). Die RGR der Linien korrelierte mit der RGR der Testkreuzungen (r = 0.42), hatte jedoch keinen Einfluss auf deren Gesamttrockenmasse. Ein Satz von 375 EU-F, EU-D und NA-D Linien wurde unter kühlen (16/13°C) und optimalen (27/25°C) Temperaturen in Klimakammern untersucht. Die photosynthetische Leistung und die Jugendentwicklung wurden für jedes Verfahren gemessen. Aus dem Verhältnis der Leistungen unter kühlen und optimalen Bedingungen wurde ein Adaptations-Index (AI) berechnet. Neunzehn Genorte (QTL) wurden für verschiedene Merkmale und deren AI mittels GWA identifiziert. Gene mit Beteiligung in der Äthylen-Signalkette, Brassinolid- und Lignin-Biosynthese wurden als Kandidaten identifiziert. Mehrere QTL für photosynthetische Leistung co-lokalisierten mit bereits beschriebenen QTL. Der Vergleich der genetischen Varianzen zeigte, dass diese innerhalb der einzelnen Landsorten grösser ist als innerhalb des EU-F Zuchtmaterials. Sowohl die hohe mittlere genetische Distanz zwischen den DH-Linien einer Landsorte, als auch das rasch abfallende Kopplungs-ungleichgewicht innerhalb der Landsorten deuten auf eine grosse Effektive Populationsgrösse hin. Die erwartete Eliminierung von rezessiven letalen Allelen durch die DH-Methode konnte den Ertrag synthetischer Landsorten nicht erhöhen und hatte auch keinen grossen Einfluss auf die genetische Diversität. Mittels GWA Analyse in den DH-Linien aus Landsorten und in Elite-Inzuchtlinien konnten 49 QTL für 27 Merkmale kartiert werden. Eine grössere Anzahl von DH-Linien aus Landsorten würde es erlauben, die durch Populationsstruktur verursachten Artefakte zu beseitigen und somit die Wahrscheinlichkeit, neue Allele zu entdecken, stark erhöhen. Zusammengefasst kann die genetische Diversität des Zuchtmaterials durch die Einkreuzung von DH-Linien aus Landsorten stark erhöht werden. Der grosse Abstand zwischen der Leistung des Zuchtmaterials und den DH-Linien aus Landsorten (22%) kann durch Selektion der besten DH-Linien teilweise ausgeglichen werden. Marker-gestützte Selektion könnte das Einkreuzen von positiven QTL ohne Introgression von unerwünschten negativen Eigenschaften erleichtern.

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    Authors: Imran, Muhammad;

    Seed reserves are the primary source of mineral nutrients for early seedling development. ?Nutrient seed priming? is a technique in which seeds are soaked in nutrient solution and subsequently dried back to initial moisture content for storage. It is an efficient approach to increase seed nutrient contents along with priming effects to improve seed quality, germination speed and seedling establishment. Various abiotic stresses, such as sub-optimal temperature, drought, submergence and soil pH extremes can seriously affect seedling establishment and nutrient acquisition at early growth stages. This thesis focused on the role and contribution of nutrient seed priming in plant growth and nutritional status in maize and soybean under conditions of limited nutrient availability and low root-zone temperature. Protocols for nutrient seed priming with Zn, Mn, Zn+Mn, B and P were optimised for application in maize and soybean seeds (B and P priming in maize only). Optimum priming durations of 24 h (maize) and 12 h (soybean) were identified for both plant species but in instead of submerging seeds in priming solutions slow imbibition between filter papers was essential for soybean to minimise development of abnormal seedlings to avoid imbibition damage. Nutrient concentrations were calculated according to water uptake to double the natural seed reserves of the respective micronutrients and 50% increase in phosphorus. However, final uptake of the micronutrients was generally much higher (+500-1000%) while it was lower for P (+20%). In case of soybean this could be attributed to a high Zn and Mn binding capacity of the seed coat, which adsorbed up to 60% of the primed nutrients. Particularly, Zn and Zn+Mn priming stimulated plant growth in hydroponic culture systems and to a lower extent also on a soil with low availability of P, Zn and Fe. This was associated with a high shoot translocation of the primed nutrients (Zn and Mn), which was most expressed in the hydroponic culture system. Combined priming treatments with Zn and Mn were usually less effective than Zn priming alone, suggesting an antagonistic interaction. By contrast, mobility of primed B was extremely low and B priming was completely ineffective. In soil culture also P priming moderately increased shoot biomass production by 10-20 %. However the efficiency of P priming was largely limited by the high P demand of the plants. Low root zone temperature (RZT) at early spring is a limiting factor for maize production in Central and Northern Europe. Nutrient acquisition, nutrient uptake and particularly root growth are severely affected at low RZT and the consequences of these growth depressions are often not completely compensated until final harvest. Model experiments in hydroponics and soil culture revealed that maize nutrient seed priming with Zn, Mn and Fe is a promising strategy to diminish the deficiency of specific nutrients, such as Zn, Mn and also P and to maintain plant growth under low RZT stress. This was mainly attributed to significantly increased root growth and particularly fine root production in plants grown from nutrient-primed seeds. Improved net photosynthesis of primed plants was mainly related with increased leaf area and preliminary results suggest a higher tolerance to oxidative damage due to increased production of protective phenolics. Two independent field experiments under conditions of suboptimal temperatures during germination and early growth revealed an increase in grain yield of 10 ? 15 % for plants derived from Zn+Mn and Fe primed seeds. This finding demonstrates long-lasting persistence of priming effects. The molecular and physiological mechanisms behind require further investigation. Samenreserven sind die primäre Quelle für die Mineralstoffversorgung während der frühen Keimlingsentwicklung. Nährstoffpriming ist ein methodischer Ansatz, bei dem Saatgut in Mineralstofflösungen vorgequollen und anschließend zur weiteren Lagerung auf Ausgangsfeuchte zurückgetrocknet wird. Hierdurch ist es möglich, neben vorquellungsbedingten Primingeffekten auch die Mineralstoffreserven zur Verbesserung der Saatgutqualität zu erhöhen sowie die Auflauf-Geschwindigkeit und die Keimlingsentwicklung zu verbessern, die durch verschiedenste Stressfaktoren, wie suboptimale Temperaturbedingungen, Trockenheit, Staunässe und pH-Extreme beeinträchtigt werden kann. Die vorliegende Arbeit beschäftigt sich mit der Wirkung von Nährstoffpriming auf das Wachstum und den Ernährungsstatus von Mais und Soja unter Bedingungen eingeschränkter Nährstoffverfügbarkeit und niedriger Wurzelraumtemperatur. Für die Saatgutapplikation von Zink (Zn), Mangan (Mn), Mn+Zn, Bor (B), Eisen (Fe) und Phosphat (P) bei Mais und Soja wurden Primingprotokolle entwickelt. Bei beiden Pflanzenarten ergab sich eine optimale Behandlungsdauer von 24 h. Jedoch war es notwendig, das Soja-Saatgut anstelle von submerser Inkubation in den Nährlösungen, langsam in Nährlösungsgetränktem Filterpapier einzuquellen, um die Entwicklung abnormaler Keimlinge aufgrund von Quellungsschäden zu vermeiden. Anhand der Wasseraufnahme der Keinlinge wurden die Nährlösungskonzentrationen so berechnet, das sich nach Einquellen eine Verdopplung der natürlichen Mineralstoffreserven im Samen ergeben sollte. Jedoch wurden für die Mikronährstoffe erheblich höhere Aufnahmeraten gemessen (+500 - 1000%), während die Aufnahme von P geringer war (+ 20%). Bei Soja konnte gezeigt werden, das die erhöhten Mikronährstoffgehalte auf eine hohe Bindungskapazität der Zwei unabhängige Feldversuche bei suboptimaler Temperatur während der Keimphase und des Jugendwachstums ergaben Kornertragssteigerungen von 10-15 % bei Mais nach Saatgutpriming mit Zn+Mn oder Fe. Diese Ergebnisse belegen die Langzeitwirkung der Primingeffekte. Die molekularen und physiologischen Grundlagen erfordern allerdings noch weitere Untersuchungen.

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    Authors: Kirschner, Stefanie;

    Industrielle Röntgendurchleuchtungsgeräte lassen sich für die Untersuchung verschiedenster Fragestellungen anwenden. Das Spektrum der Anwendbarkeit reicht von der hochauflösenden in vivo-Bildgebung von z. B. Glioblastomen, zerebralen Hirngefäßen, abdominellen und thorakalen Organen in der Maus, bis hin zur hoch-auflösenden Untersuchung von Objekten wie z. B. Stents, Zähnen, Insekten oder auch einzelner Organe ex vivo. Desweitern zeichnet das hier eingesetzte industrielle Röntgengerät die Möglichkeit zur Strahlentherapie von Tieren aus, was mit herkömmlichen speziellen Kleintier-Computertomographen nicht möglich ist. Ziel dieser Dissertation war zum einen die Etablierung eines Glioblastom-Maus-Modells und anschließend die Überprüfung der Anwendbarkeit eines Mikro-CT für die wiederholte bildgebende Darstellung der Hirntumoren vor und nach Strahlentherapie. Darüber hinaus wurde die Mikro-CT für die Evaluierung sowohl der Verschlussrate von Aneurysmen als auch der artefaktfreien Darstellung der verwendeten Materialien im Modellversuch getestet. Wir konnten in beiden von uns durchgeführten Studien die Mikro-CT erfolgreich zur Bildgebung anwenden. Zur Etablierung des Glioblastom-Maus-Modells wurde die gesamte OP-Methode zur Tumorzellimplantation in das Gehirn zunächst an toten Mäusen durchgeführt. Anstatt Zellen wurde eine äquivalente Menge an mit Kontrastmittel vermengter Methylcellulose des später geplanten Injektionsvolumens an Zellen in das Gehirn appliziert. Anschließend wurde zur Erfolgskontrolle eine Röntgenaufnahme im Mikro-CT angefertigt. Anhand der Aufnahmen konnten wir die genaue Lokalisation und Ausdehnung der applizierten Menge beurteilen und unsere OP-Methode gegebenenfalls optimieren. Erst nach verlässlicher Reproduzierbarkeit der OP wurden humane Zellen in vivo in die rechte Hemisphäre von insgesamt 27 NOD/SCID/yc-/- Mäusen injiziert. Zwei Wochen nach Xenotransplantation erfolgte eine erste kontrasmittelgestützte Untersuchung im Mikro-CT zur Tumordetektion. Bei negativem Befund wurde bei den Tieren wöchentlich eine weitere Mikro-CT durchgeführt. Tiere mit positivem Befund wurden je nach geplantem Versuch nicht therapiert (n=21) oder mittels Strahlentherapie (n=6) behandelt. Alle Tiere wurden im weiteren Verlauf wöchentlich bis zum Versuchsende im Mikro-CT untersucht. Insgesamt wurden 103 kontrastmittelgestützte Untersuchungen im Mikro-CT durchgeführt. Der maximale Tumordurchmesser und das maximale Tumorvolumen wurden anhand der einzelnen CT-Datensätze bestimmt. Zusätzlich wurden die Gehirne histologisch aufgearbeitet um das maximale Volumen und den maximalen Durchmesser der Tumore anhand der Histologie zu berechnen. Die histologischen Ergebnisse wurden mit der jeweils letzten Mikro-CT (durchgeführt am Tag des Versuchsendes) verglichen. Die hervorragende Korrelation von maximalem Tumordurchmesser und Tumorvolumen ermittelt in der Mikro-CT mit den Ergebnissen aus der Histologie (Tumordurchmesser: 0,929, p < 0,001; Tumorvolumen: 0969, p < 0,001, n = 17) belegt die hohe Genauigkeit und damit Anwendbarkeit der Mikro-CT für ein Hirntumormodell. Die hohe Sensitivität (0,95) und Spezifität (0,71) belegen zusätzlich die Exaktheit der Mikro-CT zur Erkennung von Gehirntumoren in sehr kleinen Tieren wie Mäusen (PPV=0,91, NPV=0,83). Die zweite im Rahmen dieser kumulativen Dissertation eingereichte Arbeit befasst sich mit der Evaluation einer neuen Methode zur Embolisation von Aneurysmen. Hierzu wurden Aneurysmen mit einem dicht gewobenem Stent (sog. Flow diverter) überdeckt. Über einen zuvor im Aneurysma platzierten Mikrokatheter wurde das Aneurysma mit Mikropartikeln embolisiert, welche größer als die Maschen des flussmodulierenden Stents sind. Die Mikro-CT konnten wir erfolgreich einsetzen, um mit einer hohen Auflösung die 700–900 µm kleinen Partikel im Aneurysma darzustellen und die komplette Ausfüllung des Aneurysmas zu verifizieren. Auch die Maschendichte des Stents im Verhältnis zum Aneurysma konnte mittels Mikro-CT analysiert werden. Die Möglichkeit der wiederholten Darstellbarkeit von Gehirntumoren im zeitlichen Verlauf mittels Mikro-CT bietet eine Grundlage für weitere Versuchsvorhaben, wie z. B. die Visualisierung morphologischer Veränderungen des Tumors unter verschiedenen Therapieansätzen. Die erfolgreiche Evaluation der Verschlussrate von Aneurysmen mittels einer neuen Technik in der Mikro-CT ist ein wesentlicher Schritt, bevor es zu einer ersten Anwendung im lebenden Tier kommt. Beide hier vorgestellten Arbeiten tragen durch den erfolgreichen Einsatz eines bildgebenden Verfahrens erheblich zur langfristigen Senkung der Versuchstierzahlen bei. Zusammenfassend konnten wir durch die erfolgreiche Anwendung der Mikro-CT in den hier vorgestellten Arbeiten das Spektrum der Anwendbarkeit eines industriellen Mikro-CTs in der präklinischen Forschung deutlich erweitern. Micro-CT systems are being used with increasing frequency in preclinical research. The range of examinations includes high-resolution in vivo imaging of glioblastoma, cerebral vessels, abdominal and thoracic organs in mice as well as investigation of other small objects like stents, teeth, insects or single organs ex vivo or in vitro. In addition, micro-CT may be used for radiation therapy of small rodents like mice and rats, as shown in our studies. Aim of this thesis was to establish an orthotopic glioblastoma mouse model and to evaluate the applicability of a micro-CT for repetetive imaging of brain tumors in mice. We furthermore used a micro-CT to evaluate the distribution of microspheres used to treat a silicone aneurysm model. The glioblastoma mouse model was trained in dead mice first. Instead of tumor cells we first injected methylcellulose mixed with contrast agent to train intracerebral injections. The use of contrast agent allowed us to verify our injection technique by subsequent micro-CT. After successfully establishing the technique, human brain tumor cells were in injected into the right hemisphere of n = 27 NOD/SCID/yc-/- mice. Two weeks after xenotransplantation mice underwent first contrast-enhanced micro-CT. Animals with negative results in micro-CT received additional micro-CT scans weekly until brain tumors were detected. Animals were left untreated (control group), or were treated with radiation- or chemotherapy. During treatment all animals received weekly micro-CT scans until animals had to be sacrificed. The maximum diameter and volume of intracerebrally growing xenografts were measured. Brains of sacrificed mice were histologically worked up and maximum tumor diameter and tumor volume were measured again. Results from histology were compared to micro-CT data. The excellent correlation between maximum tumor diameter and tumor volume measured in micro-CT and histology (tumor diameter: 0.929, p < 0.001; tumor volume: 0.969, p < 0.001, n=17). Sensitivity and specificity of micro-CT were 0.95 and 0.71, respectively (PPV=0.91, NPV=0.83). This proved the high accurary of micro-CT for brain tumor detection in animals as small as mice. The second paper referenced to within this doctoral thesis describes the evaluation of the distribution of microspheres within a silicon aneurysm and the degree of aneurysm occlusion. For this purpose a silicon aneurysm was treated with a densely woven stent (a so called flow diverter). Additionally, the aneurysm was embolized with microspheres larger than the pores of the stent via a microcatheter previously placed in the aneurysm. Using our micro-CT we were able to visualize the 700-900 µm small microspheres, which allowed us to verify complete embolization of the aneurysm. Furthermore, the mesh density of the flow diverter in relation to the size of the microspheres was analyzed using the micro-CT. From our findings we conclude, that micro-CT is a versatile tool that may be used for imaging of a vast range of objects and thus propose that micro-CT will be used with increasing frequency in preclinical research.

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    Authors: Krishnan, Parvathy;

    Pathogenic fungi impose a great threat to the global food production. They are globally distributed and display a huge amount of phenotypic diversity. They demonstrate high adaptability to diverse environments and to fluctuating conditions within their hosts. However, not much is known about the mechanisms underlying these processes of adaptation. Deciphering the genetic basis of phenotypic diversity may help us to get a deeper insight into the mechanisms that regulate complex adaptive traits. Furthermore, these findings may help in improving strategies for an effective and sustainable management of pathogens. In my PhD study, I aimed to understand the genetic basis of adaptation to variable environments using Zymoseptoria tritici. Z. tritici causes septoria leaf blotch of wheat and is considered to be the most devastating fungal wheat pathogen in Europe. In the first chapter of my thesis, I studied the role of melanin in adaptation of Z. tritici. I, demonstrated that melanin is essential for protection against harmful chemical fungicides such as bixafen. Remarkably, I observed high phenotypic variability in melanin accumulation levels in different Zymoseptoria tritici strains, most probably reflecting its role in adaptation to variable environments. Using genetic mapping, I was able to demonstrate that variability in melanization levels between two Swiss strains of Z. tritici was mediated by differential expression of the transcription factor Zmr1 (Zymoseptoria melanin regulation 1). I identified nucleotide mutations in the promoter region and an insertion of transposable elements (TEs) upstream of the promoter region as two mechanisms responsible for this observed differential expression of Zmr1. My findings clearly demonstrated the significance of gene expression regulation, mediated by the insertion of TEs and SNPs, in optimizing the growth and adaptation of the fungus under variable environments. Furthermore, these adaptive changes are responsible for the phenotypic diversity that we observe in Z. tritici strains from all over the world. In the second part of my thesis, I analyzed the expression of several secreted peptidases of Z. tritici at distinct stages during the infection process of wheat plants. Using computational genetic methods, I also analyzed nucleotide data sets of the peptidase genes for evolutionary signatures of genetic adaptation. By combining both approaches, I aimed at obtaining a more detailed picture of the relevance and the role of each enzyme during the infection process. I was able to show that the peptidases belonging to MEROPS families A1 and G1 were significantly up regulated during the asymptomatic phase of infection and displayed signatures of accelerated evolution, suggesting their key role in suppression of plant defenses and host specialization. In the third part of my thesis, I evaluated the role of a cellulase-encoding gene (JGI Prot. ID: 76589) using molecular biology techniques. This cellulase was only expressed during the final necrotrophic phase of Z. tritici infection and the nucleotide sequences of the gene showed signatures of diversifying selection. This pattern is consistent with the hypothesis of evasion from host recognition during the first phases of infection and subsequent induction of host-cell death by the secretion of an array of different cellulase isoforms by the fungus. To test this hypothesis, I constitutively expressed the cellulase-encoding gene in Z. tritici during all infection stages by inserting a constitutive promoter (originally from the Glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans) upstream of the start codon of the cellulase gene. I was able to show that the progress of infection in this mutant was delayed compared to the non-modified Z. tritici isolates. This suggests that the cellulase acts as an elicitor molecule that is detected by the plant and triggers the defense machinery that will hinder the growth of the pathogen. In summary, my PhD work demonstrated the significance of fine-tuning of gene expression regulation of various genes in plant pathogenic fungi to facilitate adaptation to different environmental conditions and host colonization. Remarkably, I was able to show that gene expression fine-tuning resulted in large amounts of phenotypic diversity in adaptive traits.

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    ETH Zürich Research Collection
    Doctoral thesis . 2018
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      ETH Zürich Research Collection
      Doctoral thesis . 2018
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    Authors: Šumrada, Tanja;

    Kmetijstvo sodi med najpomembnejše dejavnike zmanjševanja biotske pestrosti v EU, vendar integracija varstva narave v Skupno kmetijsko politiko ostaja nezadostna. Namen doktorske disertacije je proučiti načrtovanje, učinke in vrednotenje slovenske kmetijske politike na področju ohranjanja narave. Analiza z modeli BRT je pokazala negativno povezavo med diverziteto ptic kmetijske krajine in neposrednimi plačili, okoljski ukrepi pa so imeli v obdobju 2008–2019 na diverziteto ptic šibek relativni vpliv. Diverziteta ptic je bila najvišja v odprti krajini na območjih Natura 2000 z visoko pestrostjo kmetijskih rastlin in nizko obtežbo z živino. Programsko-teoretična analiza slovenske kmetijske politike je pokazala številne vrzeli v opredelitvi intervencijske logike. Manjka predvsem opredelitev ciljev in kazalnikov, ki bi omogočali presojo uspešnosti ukrepanja. Integracija varstva narave je na strateškem nivoju omejena predvsem na prostovoljne ukrepe, strategije za preprečevanje konfliktov med naravovarstvenimi in drugimi cilji kmetijske politike pa večinoma niso opredeljene. Ocenili smo, da je bilo v programskem obdobju 2014–2020 varstvu narave namenjenih okrog 5 % (17,8–19,4 mio EUR) letnega proračuna slovenske kmetijske politike. Disertacijo zaokrožuje analiza možnosti za izboljšanje rezultatske naravnanosti kmetijsko-okoljskih ukrepov, ki je potekala s poskusom diskretne izbire (521 kmetov na območju Haloz in Krasa) ter kvalitativno tematsko analizo intervjujev s kmeti, raziskovalci, odločevalci in kmetijskimi svetovalci. Kmetje so izkazali statistično značilno večje preference do rezultatskih shem kot do shem s predpisanimi praksami ter naklonjenost do individualnega pristopa k svetovanju. Za uspešno uveljavitev rezultatskih shem je nujno izboljšanje podatkovnih podlag, raziskav trajnostnih proizvodnih modelov ter usposobljenosti kadrov v nadzornih in svetovalnih službah. Agriculture is one of the most important factors in reducing biodiversity in the EU. However, the integration of nature conservation into the Common Agricultural Policy remains inadequate. The purpose of the doctoral dissertation is to study the planning, effects and evaluation of the Slovenian agricultural policy in the field of nature conservation. An analysis using BRT models showed a negative link between the diversity of farmland birds and direct payments, while environmental measures had a weak relative influence on bird diversity in the period 2008–2019. Bird diversity was highest in open landscapes on Natura 2000 sites that had a high diversity of agricultural plants and low stocking density. A programme theory analysis of the Slovenian agricultural policy showed numerous gaps in the definition of the intervention logic. Particularly lacking is a definition of objectives and indicators that would enable an assessment of the effectiveness of interventions. At the strategic level, the integration of nature conservation is limited mainly to voluntary measures. Strategies for preventing conflicts between nature conservation and other objectives of agricultural policy are mostly not defined. We estimated that in the 2014–2020 programming period, approximately 5 % (17.8–19.4 million EUR) of the annual budget of the Slovenian agricultural policy was allocated to nature conservation. The dissertation is concluded with an analysis of the possibilities for improving the result-orientation of agri-environmental measures, which was conducted with a discrete choice experiment (521 farmers in the Haloze and Karst area) and a qualitative thematic analysis of interviews with farmers, researchers, decision-makers and agricultural advisers. The farmers showed statistically significant higher preferences for result-based schemes than for schemes with prescribed practices and a preference for an individual approach to knowledge transfer. In order to successfully implement result-based schemes, it is necessary to improve data bases, research into sustainable production models and the training of staff in supervisory and advisory services.

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    Authors: Wapelhorst, Britta;

    In der vorliegenden Doktorarbeit sollte a) die Expression sowie b) die zelluläre Lokalisation des SLC10A6 (SOAT) im humanen Hoden geklärt werden. Der SOAT zeigt im Vergleich mit weiteren Gewebetypen vergleichsweise hohe Expression im Hoden und weist eine Transportaffinität für sulfatierte Steroide auf. Sulfatierte Steroide können aufgrund ihrer hydrophilen Eigenschaften Zellmembranen nicht durch Diffusion passieren, sondern benötigen einen Transporter. Zudem sind sulfatierte Steroide biologisch nicht aktiv und können erst nach Desulfatierung an ihre entsprechenden Rezeptoren binden und eine biologische Antwort hervorrufen. Aufgrund seiner Eigenschaften ist der SOAT ein potentieller Kandidat für den Transport sulfatierter Steroide im humanen Hoden und somit für die lokale Bereitstellung von Androgenen und Östrogenen. Die Untersuchungen wurden an Hodenbiopsien von Patienten durchgeführt, die eine normale Spermatogenese aufwiesen und Hodenbiopsien von Patienten mit gestörter Spermatogenese, bis hin zu einer völligen Abwesenheit von Keimzellen, dem Sertoli Cell Only Syndrom. Der Nachweis der Expression des SOAT erfolgte mittels TaqMan®-RT-qPCR, RT-PCR nach Laser-assistierter Mikrodissektion und In-situ- Hybridisierung. In den Untersuchungen der vorliegenden Arbeit konnte durch TaqMan®-RT-qPCR und RT-PCR nach Laser-assistierter Mikrodissektion gezeigt werden, dass der SOAT in Hodenbiopsien mit normaler Spermatogenese exprimiert wird. Der zelluläre Nachweis des SOAT erfolgte mittels In-situ-Hybridisierung in Keimzellen, den primären Spermatozyten. Für den Nachweis des SOAT Proteins wurden im Rahmen dieser Dissertation sechzehn Antikörper eingesetzt. Eine spezifische Färbung konnte jedoch bei keinem Antikörper beobachtet werden. Untersuchungen von Fietz et al. (2013) konnten das SOAT-Protein schließlich mittels eines Antikörpers gegen das Soat-Protein der Maus Soat(m329-344) in Spermatozyten und teilweise in Spermatiden nachweisen. Da es sulfatierten Steroiden nicht möglich ist, die Blut-Hoden-Schranke ohne die Hilfe eines Transporters zu überwinden und so die Keimzellen zu erreichen, stellt sich die Frage nach der Funktion des SOAT in den Keimzellen des humanen Hodens. Der Nachweis der Expression des SOAT erfolgte auch in drei Hodenbiopsien mit Spermatogenestörungen, die mit einer Infertilität einhergehen. Aufgrund der signifikant geringeren Expression des SOAT in Hodenbiopsien mit einer Hypospermatogenese kann vermutet werden, dass eine signifikant geringere Expression des SOAT mit einem verringertem Transport von sulfatierten Steroiden und in Folge mit einer reduzierten Bereitstellung von lokal verfügbaren Androgenen und Östrogenen einhergeht (Fietz et al. 2013). Diese Fragestellung könnte durch Untersuchungen der Spermatogenese von Slc10a6-knockout Mäusen weiterführend untersucht werden. It was the aim of this doctoral thesis to elucidate a) the expression and b) the cellular localization of the sodium-dependent organic anion transporter SLC10A6 (SOAT) in human testis. SOAT is predomiantly expressed in human testis and shows specific transport for sulfated steroids. Because of their hydrophilic nature, sulfated steroids are not able to pass cell membranes and therefore need the help of a transporter. Moreover, sulfated steroids are not biologically active, only after desulfatation they can bind to their receptors to induce a biological response. SOAT is a potential candidate not only for transport of sulfated steroids in the human testis, but also for a local supply of androgens and estrogens. The study included testicular biopsis showing normal spermatogenesis and spermatogenic impairment, including Sertoli cell only syndrome. Detection of SOAT mRNA was performed by TaqMan®-RT-qPCR, RT-PCR after laser-assisted microdissection and in situ hybridization. TaqMan®-RT-qPCR and RT-PCR after LAM detected expression of SOAT in testicular biopsies showing normal spermatogenesis. SOAT was localized in germ cells, in primary spermatocytes by in situ hybridization. For verification of SOAT protein distribution 16 antibodies were tested but none could give answer to the question of protein localization. Fietz et al. (2013) employed a new antibody against SOAT protein of mice (Soatm329-433) and detected SOAT in spermatocytes and spermatid of various stages. Due to unabillity for sulfated steroids to pass the blood-testis-barrier, the function of SOAT in spermatocytes is still not known. SOAT could be verified in three testicular biopsies showing impaired spermatogenesis resulting in infertility. Fietz et al. confirmed a significant lower expression of SOAT in testicular biopsies showing hypospermatogenesis. It is suggested that the reduction or lack of SOAT expression might be related to a decreased local supply with androgens and estrogens (Fietz et al. 2013). Further studies with Slc10a6-knockout mice could corroborate this hypothesis.

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    Authors: Kumar, Sandhya Sasi;

    Since its founding as a social welfare state, India has been meandering through “a life of contradictions” when it comes to social and economic equality. A potent indicator of this reality is the state of food and nutrition security across the country. India’s meager progress comes on the heels of rapid economic growth over the last two decades and a slew of public programs have attempted to address the multi-faceted nature of food and nutrition security. These efforts have included subsidized grains for households through the Targeted Public Distribution System (TPDS), village health, nutrition and education programming through the Integrated Child Development Services (ICDS), and Nutrition Rehabilitation Centers (NRCs) to treat cases of severe acute malnutrition (SAM). Though these programs have been scaled across the country, numerous governance challenges and institutional failures have persisted. The latest development in India’s food and nutrition policy landscape is the National Food Security Act (hereafter referred to as the Act) of 2013, which leverages existing national programs, including the TPDS and ICDS, to grant legal entitlements to nearly 70% of the total population. This Act was the fruit of a rights-based movement in India that demanded for over a decade to codify social and economic rights. However, the question arises as to whether this new Act been able to achieve its goal to ensure food and nutrition security by strengthening the governance of these programs? This is the central question of the present thesis. This thesis investigates how the Act has reformed the implementation of food and nutrition programs in the states of Jharkhand, Madhya Pradesh and the capital of New Delhi. The four key objectives of this study are to investigate 1) what changes are prescribed by the Act; 2) how effective these reforms have been in addressing persistent governance challenges; 3) what challenges remain and why; and 4) what strategies could be used to address these gaps. A qualitative case study approach was applied, which involved the following data collection methods: key informant and in-depth interviews, focus group discussions, participant observation, matrix ranking and process net-mapping, an innovative participatory tool that maps complex institutional processes and actors. This thesis is comprised of nine chapters. The first chapter introduces the puzzle of India’s nutritional status and explores why government efforts to tackle this problem have had tepid results. The second chapter presents the conceptual framework that defines governance challenges and an institutional framework of demand and supply side factors that contribute to effective policy and programming. Chapters three and four present an overview of the TPDS, ICDS and NRCs, and review the existing literature on the governance issues of these programs. Chapter 5 lays out the study design, methods and ethical protocols. Chapters six through eight explore each of the three programs and present key findings across the selected study sites. The last chapter presents cross-cutting conclusions for these programs in the wake of the Act and proposes a way forward to address institutional gaps in tackling food and nutrition security. Seit der Gründung des Wohlfahrtsstaates erlebt Indien eine Reihe von Widersprüchen, was die soziale und wirtschaftliche Gerechtigkeit des Landes angeht. Ein aussagekräftiger Indikator dieser Realität ist der Stand der Ernährungsicherheit im Land. Indiens dürftiger Fortschritt in der Ernährungssicherung folgt einer zwei Jahrzehnte andauernden Periode rasantem Wirtschaftswachstum, sowie einer Reihe von öffentlichen Programmen, die versuchen, die vielschichtigen Aspekte von Ernährung zu adressieren. Zu diesen Bemühungen gehören subventioniertes Getreide für Haushalte durch das gezielte öffentliche Vergabesystem TPDS, dörfliche Gesundheits-, Ernährungs- und Bildungsprogramme durch die Integrierten Dienstleistungen zur Entwicklung von Kindern ICDS und Rehabilitierungszentren für Fälle von schwerer akuter Mangelernährung (NRCs). Obwohl diese Programme auf ganz Indien ausgeweitet wurden, bestehen weiterhin institutionelle Mängel sowie zahlreiche Governance-Herausforderungen. Die jüngste Entwicklung in der Indischen Ernährungspolitik ist das Gesetz zur nationalen Ernährungssicherheit NFSA aus dem Jahr 2013. Das NFSA nutzt zwei bestehende nationale Programme, TPDS und ICDS, um fast 70% der Bevölkerung Rechtsansprüche zu gewähren. Dieses Gesetz war das Ergebnis einer größeren rechtebasierten Bewegung in Indien, die seit über einem Jahrzehnt das Ziel hat, soziale und wirtschaftliche Rechte zu kodifizieren. Es stellt sich die Frage, ob dieses neue Gesetz es ermöglicht hat, durch eine Stärkung der Governance der bestehenden Programme einen Lebenszyklusansatz in der Ernährungssicherung zu etablieren. Diese Doktorarbeit verwendet einen Ansatz vergleichender Fallstudien, um zu untersuchen, wie das NFSA die Umsetzung der Ernährungsprogramme in den Staaten Jharkhand, Madhya Pradesh und der Hauptstadt Neu Delhi reformiert hat. Die vier Hauptziele der Dissertation sind: 1) die Änderungen, die durch das NFSA vorgeschrieben wurden, zu beschreiben 2) die Effektivität dieser Reformen bei der Bewältigung von Governance-Herausforderungen zu erklären, 3) die weiterhin bestehenden Governanceprobleme und Gründe dafür zu erklären, und 4) Strategien zu erarbeiten, die zur Behebung dieser Defizite beitragen können. Diese vier Aspekte wurden untersucht mittels Befragung von Schlüsselpersonen, Tiefeninterviews, Fokusgruppendiskussionen, teilnehmender Beobachtung, Matrix-Ranking und Prozessnetzkartierung (process net-mapping), ein innovatives und partizipatives Instrument, das komplexe institutionelle Prozesse und Akteure abbildet. Die vorliegende Doktorarbeit besteht aus neun Kapiteln. Das erste Kapitel gibt eine Einleitung in die Rätselhaftigkeit der Ernährungslage in Indien, und warum staatliche Bemühungen dieses Problem anzugehen, nur dürftige Ergebnisse erzielt haben. Das zweite Kapitel stellt einen konzeptionellen Rahmen bezüglich Governance-Herausforderungen und einen institutionellen Rahmen bezüglich Nachfrage- und Angebotsfaktoren, die zu wirkungsvollen Politikmaßnahmen und Programmgestaltung beitragen, dar. Kapitel drei und vier geben einen Überblick über TPDS, ICDS und NRCs, und über die bestehende Literatur bezüglich der Governance-Herausforderungen dieser drei Programme. Kapitel fünf beschreibt das Studiendesign, die angewandten Methoden und die Forschungsprotokolle. Kapitel sechs bis acht untersuchen jeweils eins der drei Programme und erklären Schlüsselergebnisse über die drei ausgewählten Untersuchungsstandorte hinweg. Die Dissertation schließt mit einer übergreifenden Zusammenfassung für diese Programme vor dem Hintergrund des neuen Gesetzes und gibt wegweisende Vorschläge wie man diese institutionellen Defizite zur Verbesserung der Ernährungssicherheit adressiert.

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    Authors: Yilmaz, Gülenay;

    The fact that rapid diagnosing of especially seed-borne pathogens and their control is the first step to obtain healthy crop. The aims of the study were to optimize the analyzing methods in detecting presence of Plasmopara halstedii (Farl.) Berl. & De Toni requiring analysis at the time of importation in the sunflower seed using current molecular methods that were much more advantageous than that of classical methods, and to evaluate potential variations in sensitivities of the pathogen to metalaxyl and mefenoxam. Calculations related to classical PCR (agarose gel electrophoresis) were optimized by referring to Appendix 2 of the EPPO Standard Protocols while those related to real-time PCR (the hydrolyze probes) were optimized by referring to Appendix 3 of the EPPO Standard Protocols. Moreover, SyberGreen Mastermix Kit protocol was used for optimization for the study of real-time PCR (SyberGreen) that is not present in the EPPO Standard Protocols. Thus, it was provided that all three molecular protocols were confirmed for better applicable and practical ways for the quarantine laboratories in our country. Furthermore, the first molecular studies on direct detection of the pathogen from sunflower seeds through these optimization was conducted, as well. Reduced sensitivities of P. halstedii to metalaxyl are well-known throughout the world. In this study, it was showed that the similar reduced sensitivities of the pathogen had been determined for mefenoxam that is an isomer of metalaxyl. There were found no differences statistically and both fungicides were placed in the same group because of resulting from cross-resistance between. Özellikle, tohum kaynaklı olan ve tohumla taşınabilen etmenlerin hızlı tanılanması ve bunların kontrol altına alınması sağlıklı bir ürün elde etmenin ilk aşamasını oluşturmaktadır. Bu tez çalışmasının amacı, ithalat aşamasında analiz şartı olan tohum kaynaklı Plasmopara halstedii (Farl.) Berl. & De Toni)'nin ayçiçeği tohumundaki varlığının klasik yöntemlere göre çok daha avantajlı moleküler yöntemler kullanılarak optimizasyonunun gerçekleştirilmesi ve metalaxyl ile mefenoxam arasındaki duyarlılıklarının değerlendirilmesidir. Klasik PCR (agaroz jel elektroforez) ile ilgili hesaplamaları EPPO Standart Protokolleri Appendix 2 ve Real Time PCR (hidroliz probları) ile ilgili hesaplamalar ise EPPO Standart Protokolleri Appendix 3 referans alınarak optimize edilmiştir. Ayrıca, EPPO Standart Protokollerinde yer almayan Real time PCR (SyberGreen) çalışmasında ise, SyberGreen Mastermix kit protokolü esas alınarak optimize edilmiştir. Böylece, ülkemiz karantina laboratuarları içerisinde her üç moleküler protokülün daha uygulanabilir olması sağlanmıştır. Ayrıca, bu optimizasyon ile etmenin ayçiçeği tohumundan direkt tespitine yönelik ilk moleküler çalışma da gerçekleştirilmiştir. P. halstedii izolatlarının metalaxyl'e duyarlılıklarının azaldığı tüm dünyada bilinmektedir. Bu çalışmada da, söz konusu patojenin metalaxyl'in izomeri olan mefenoxam'a karşı da duyarlılıklarının azaldığı belirlenmiş ve aralarında çapraz dayanıklılık olduğu için metalaxyl ile aynı grupta yer almıştır.

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    Authors: Hailu, Teweldemedhn Gebretinsae;

    Ethiopia is a major beekeeping country located in northeast Africa where several evolutionary lineages of Apis mellifera contact. A unique practice of honey bee colony marketing which involves broad agro-ecological zones (AEZs) is a developing trend in the northern part of the country such as Tigray region in association with apicultural development. Several studies based on classical morphometry on the Ethiopian honey bee subspecies classification debated from the unique Apis mellifera simensis to five others. Moreover, the genetic diversity, adaptation, gene flow and inter-relationships of the honey bees between AEZs were not disentangled – a challenge for planning sustainable apicultural development and conservation. Therefore, this study was conducted to elucidate the honey bees of Ethiopia in a context of apicultural transformation using integrated methods: morphometrics, genetics, colony market survey and metadata analyses on beekeeping development. The results of geometric morphometric analyses confirmed that Ethiopian honey bees represented by Apis mellifera simensis references belong to a separate lineage (Y) compared to A, O, M and C, and the present sample belonged to Y. This supported the hypothesis of five major honey bee lineages of the honey bee Apis mellifera. Similarly, a maximum likelihood phylogenetic tree analysis based on the mitochondrial COI-COII showed that most of the Ethiopian honey bees belong to lineage Y. However, a substantial proportion of the samples from the northern part of the country clustered with lineage O, which support the hypothesis that there is close contact between Y and O. Both geometric morphometry and classical morphometry differentiated the Ethiopian honey bees from all references including A. m. monticola, A. m. scutellata, A. m. jementica, A. m. adansonii but grouped with A. m. simensis. Genetically, five DraI haplotypes (COI-COII) were found to be randomly distributed across AEZs, indicating a substantial gene flow. Consequently, the level of genetic differentiation among the Ethiopian honey bee subpopulations defined by local areas and AEZs was generally low based on r7-frag nuclear marker, which is identified to be associated with adaptation to habitat elevation in East African honey bees. Similarly, nucleotide diversity consistently decreased with increasing elevation – indicating a reduced effective population size in the highlands. Results obtained from colony market survey showed that the honey bee swarms are reproduced in a few highlands and re-distributed throughout the region. Colony buyers have preferences of color and AEZ of origin of the honey bees, which led to a one-way flow and eroded the overall level of genetic differentiation. However, a marked differentiation was detected between the highland and lowland honey bees in relic communities where an allelic length polymorphism was observed as a signature of local adaptation. Altogether, Ethiopian honey bees belong to the lineage Y and subspecies A. m. simensis, and are characterized by a high level of gene flow enhanced by colony marketing; but a conserved signature of local adaptation to higher elevations was identified in less disturbed communities. Further studies based on genome-wide analyses and field experiments, focusing on undisturbed communities, can provide more insights into adaptation, admixture and management implications. Sustainable bee breeding and extension services that enable local beekeeping without colony trade and transportation will help to promote apiculture and genetic conservation. Äthiopien ist ein Land mit vielfältigen Ökosystemen im Nordosten Afrikas, in denen viele evolutionäre Linien von Apis mellifera zusammenkommen. In der Region Tigray (im Norden von Äthiopien) werden Honigbienenvölker auf zentralen Märkten gehandelt. Mehrere Studien zur Klassifikation der Unterarten von äthiopischen Honigbienen auf der Grundlage der klassischen Morphometrie stellten unterschiedliche Hypothesen über die Anzahl dieser Unterarten auf. Darüber hinaus war der Mangel an Informationen über genetische Vielfalt, Anpassung und Genfluss eine Herausforderung für die Planung einer nachhaltigen Entwicklung der Imkerei. Ziel dieser Doktorarbeit war es, diese Honigbienen und den Handel mit ihnen auf der Grundlage von Morphometrie, Genetik und Marktuntersuchungsanalysen in einen Zusammenhang zu stellen. Die Ergebnisse bestätigten, dass äthiopische Honigbienen, die durch Apis mellifera simensis-Referenzen repräsentiert werden, von den Linien A, O, M und C morphologisch unterschieden werden können und zu der Linie Y gehören, was die Hypothese von fünf großen Honigbienen-Linien unterstützt. In ähnlicher Weise zeigte die mitochondriale COI-COII-Analyse, dass die meisten äthiopischen Honigbienen der Linie Y angehören. Ein erheblicher Anteil der Proben aus dem nördlichen Teil des Landes gruppierte sich jedoch mit der Linie O, was die Hypothese eines engen Kontaktes zwischen den Linien Y und O stützte. Hinsichtlich der Unterarten unterschieden die geometrische Morphometrie und die klassische Morphometrie die äthiopischen Honigbienen von allen Referenzen einschließlich A. m. monticola, A. m. scutellata, A. m. jementica, A. m. adansonii, aber gruppierten mit A. m. simensis. Genetisch wurden fünf DraI-Haplotypen identifiziert, die zufällig über die agro-ecological zones (AEZs) verteilt waren, was auf einen erheblichen Genfluss hinweist. Folglich war die genetische Differenzierung der äthiopischen Proben insgesamt gering. Die Abnahme der Nukleotiddiversität in den Subpopulationen von A. mellifera in höher gelegenen Habitaten deutet auf eine reduzierte effektive Populationsgröße im Hochland hin. Die Markterhebung ergab, dass sich die Honigbienenvölker in einigen Hochländern vermehren und durch die Vermarktung in der gesamten Region von Tigray verteilt werden. Eine Ursache hierfür ist, dass die Käufer die Farbe und die AEZ der Herkunft bevorzugen. Dies führte zu einem unidirektionalen Fluss von genetischem Material und verringert die genetische Differenzierung. In lokalen, ungestörten Gebieten konnte jedoch eine deutliche Differenzierung zwischen der Hochland- und der Tieflandhonigbiene festgestellt werden. Dabei wurde ein Allellängenpolymorphismus als Zeichen der lokalen Anpassung beobachtet. Zusammenfassend lässt sich feststellen, dass äthiopische Honigbienen der Linie Y und der Unterart A. m. simensis zugehörig sind. Ein hoher Genfluss zwischen Subpopulationen wird durch Koloniemarketing verstärkt; aber eine konservierte Signatur der lokalen Anpassung an höhere Lagen wurde in ungestörten Gebieten identifiziert. Weitere Forschungen auf der Grundlage genomweiter Analysen und Feldexperimente, die sich auf die ungestörten lokalen, Gebiete konzentrieren, können weitere Einblicke in Anpassung, Durchmischung und Management liefern. Nachhaltige Bienenzucht und Beratungsdienste, die eine lokale Imkerei ohne Völkerhandel ermöglichen, werden zur Förderung der Imkerei und der genetischen Erhaltung beitragen.

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    Authors: van Iersel, W.K.;

    Natural lowland rivers are dynamic environments with a high ecological value. However, 90% of the European and North-American river floodplains are in a degraded state. The functions of floodplains are strongly determined by land cover and they often compete for space in narrowed floodplains. Integrated river management (IRM) tries to take care of floodplains in such way that land cover is optimized for multiple functions. For IRM, monitoring is essential to capture the dynamics, to evaluate changes, and to document the state of floodplains over time. The main objective of this thesis was to establish remote-sensing methods for the monitoring of floodplain land cover over multiple spatial and temporal scales. Several remote-sensing based solutions have been developed for the monitoring of land-cover dynamics in river floodplains and tested in floodplains of the lower Rhine. The phenological change of floodplain vegetation over the course of one year was studied using temporal profiles of its height and greenness. Using multitemporal UAV images, vegetation height was determined with an accuracy similar to much more expensive airborne LiDAR data. Multitemporal elevation models yielded meaningful profiles of greenness and vegetation height over time, which enabled discriminating the different land-cover types. The same dataset combined with a powerful machine learning model (Random Forest) yielded unprecedented high classification accuracies for floodplain vegetation (> 90%), even for similar vegetation types such as grassland and herbaceous vegetation. This method is a practical and highly accurate solution for monitoring areas of a few square kilometres. For large-scale monitoring of floodplains, the same method is recommended, but with data from airborne platforms covering larger extents. Land-cover change over the course of five years was studied for a 100-km river section using satellite images. Using an object-based approach, a sequential deviation of a land-cover object from its class mean was used to detect land-cover change. For most classes the method was unsuccessful (accuracy 75%. The developed method has important advantages, such as high observation frequency, independence of repeated land-cover classification, and fast processing. At sub-daily frequency, it was assessed how accurate water temperature in a floodplain side channel can be documented from thermal UAV maps. The associated habitat suitability for native and alien fish assemblages was estimated based on the produced temperature maps. Water surface temperatures were mapped four times during a hot summer day with an overall RMSE of 0.53 oC. During the day, temperatures in the side channel increased rapidly to values detrimental for many fish species. The study showed that thermal imagery from UAVs is an efficient and accurate information source to monitor spatiotemporal patterns of thermal habitat suitability. The presently available range of spaceborne and airborne platforms and sensors offers great opportunities to collect information on land-cover change across a range of spatial and temporal scales. This may advance our management of floodplains and help us recovering and protecting these rich ecosystems and the benefits they provide us.

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    Authors: Strigens, Alexander Carl Georg;

    Since the introduction of maize into Europe, open-pollinated varieties of flint maize were cultivated across the continent. Natural selection promoted adaptation to the climatic conditions prevailing in the different regions. With the advent of hybrid breeding in Europe during the 1950’s, some of the genes responsible for the specific adaptations of the landraces to abiotic and biotic stress were captured in the first developed inbred lines, but most of their genetic diversity is still untapped. Development of inbred lines out of this material by recurrent selfing is very tedious due to strong inbreeding depression. In contrast, the doubled-haploid (DH) technology allows producing fully homozygous lines out of landraces in only one step. This allows their precise characterization in replicated trials and identification of new genes by genome wide association (GWA) mapping. In this study we genotyped a set of 132 DH lines derived from European Flint landraces and 364 elite European flint (EU-F), European dent (EU-D) and North-American dent (NA-D) inbred lines with 56,110 single nucleotide polymorphism (SNP) markers. The lines were evaluated in field trials for morphologic and agronomic traits and GWA mapping was performed to identify underlying quantitative trait loci (QTL). In particular, our objectives were to (1) develop a robust method for quantifying early growth with a non-destructive remote-sensing platform, (2) evaluate the importance of early growth performance of inbred lines with regard to their testcross performance, (3) determine the potential of GWA mapping to identify genes underlying early growth and cold tolerance related traits, (4) evaluate the phenotypic and genotypic diversity recovered in the DH lines derived from the landraces, (5) estimate the effect of the DH method on the recovered genetic diversity, (6) identify new genes by GWA mapping in the DH lines derived from landraces, and (8) discuss the potential of DH lines derived from landraces to improve the genetic diversity and performance of elite maize germplasm. A phenotyping platform using spectral reflectance and light curtains was used to perform repeated measurements of biomass and estimate relative growth rates (RGR) of the DH and inbred lines, as well as of two testcrosses of 300 dent inbred lines. The DH lines derived from the landraces Schindelmeiser and Gelber Badischer had the highest RGR followed by EU-F lines, DH lines derived from Bugard, EU-D lines and, finally, NA-D lines. For inbred lines, whole plant dry matter yield (DMY) was positively correlated with RGR (r = 0.49), whereas this relation was weaker in the testcrosses (r = 0.29). RGR of the inbred lines correlated with RGR of their testcrosses (r = 0.42), but it had no influence on testcross DMY. A set of 375 EU-F, EU-D and NA-D lines were further evaluated in growth chambers under chilling (16/13°C) and optimal (27/25°C) temperatures. Photosynthetic and early growth performance were estimated for each treatment and an adaptation index (AI) built as the chilling to optimal performance ratio. Nineteen QTL were identified by GWA mapping for trait performance and AI. Candidate genes involved in ethylene signaling, brassinolide, and lignin biosynthesis were found in their vicinity. Several QTL for photosynthetic performance co-located with previously reported QTL and the QTL identified for shoot dry wieght under optimal conditions co-located with a QTL for RGR. Comparison of the DH lines derived from landraces with the EU-F lines showed that genotypic variances in single DH populations were greater than in the EU-F breeding population. A high average genetic distance among the DH lines derived from the same landrace as well as a rapid decay of linkage disequilibrium suggests a high effective population size of the landraces. Because no systematic phenotypic differences were observed between the landraces and synthetic landraces obtained by intermating the corresponding DH lines, the expected purge of lethal recessive alleles during the DH production did neither improve grain yield performance nor affect the recovered genetic diversity. Performing GWA in the DH lines derived from landraces as well as the EU-F, and EU-D lines allowed the identification of 49 QTL for 27 traits. A larger set of DH lines derived from more landraces might solve problems arising from population structure and allow a much higher power for the detection of new alleles. In conclusion, the introgression of DH lines derived from landraces into the elite breeding material would strongly broaden its genetic base. However, grain yield performance was 22% higher in EU-F lines than in the DH lines derived from landraces. Selection of the best DH lines would allow partially bridging this yield gap and marker-assisted selection may allow introgression of positive QTL without introducing negative features by linkage drag. Seit der Einfuhr von Mais aus der „neuen“ Welt nach Europa, wurden offen abblühende Flint-Mais Populationen auf dem gesamten Kontinent angebaut. Durch natürliche Selektion passten sich diese Landsorten an die Klimate des Kontinents an. In den Anfängen der Hybridzüchtung wurden Gene und Allele, die für diese spezifische Anpassung an biotische und abiotische Stressfaktoren verantwortlich sind, in den ersten Inzuchtlinien nur teilweise fixiert. Der Grossteil der genetischen Vielfalt der Landsorten blieb jedoch ungenutzt, da die Entwicklung von Inzuchtlinien aus diesem Material wegen besonders starker Inzuchtdepression sehr mühsam ist. Demgegenüber erlaubt es die seit etwa 10 Jahre eingesetzte Methode der Erzeugung von Doppel-Haploiden (DH), vollständig homozygote Linien aus Landsorten in einem einzigen Schritt zu entwickeln. Diese DH-Linien können in wiederholten Feldversuchen sehr präzise evaluiert werden. Dies vereinfacht die Kartierung von Genen mithilfe der Genom-weiten Assoziations-Kartierung (GWA) enorm. In der vorliegenden Studie wurden 132 DH-Linien aus europäischen Landsorten, 364 Inzucht-linien aus Nordamerikanischem Dent (NA-D), europäischem Flint (EU-F) und europäischem Dent (EU-D) Zuchtmaterial mit 56110 genetischen Markern genotypisiert. Agronomische Eigenschaften der DH-Linien und Elite-Inzuchtlinien wurden in Feldversuchen evaluiert und mittels GWA kartiert, um vorteilhafte Gene zu identifizieren. Zu unseren Zielen gehörten insbesondere (1) die Entwicklung einer robusten, nicht-destruktiven Methode zur Erfassung der Jugendentwicklung mittels Sensoren, (2) die Untersuchung des Zusammenhangs zwischen der Jug