HelF, a suppressor of RNAi mediated gene silencing in Dictyostelium discoideum

RNA interference (RNAi) is a recently discovered process, in which double stranded RNA (dsRNA) triggers the homology-dependant degradation of cognate messenger RNA (mRNA). In a search for new components of the RNAi machinery in Dictyostelium, a new gene was identified, which was called helF. HelF is a putative RNA helicase, which shows a high homology to the helicase domain of Dicer, to the helicase domain of Dictyostelium RdRP and to the C. elegans gene drh-1, that codes for a dicer related DExH-box RNA helicase, which is required for RNAi. The aim of the present Ph.D. work was to investigate the role of HelF in PTGS, either induced by RNAi or asRNA. A genomic disruption of the helF gene was performed, which resulted in a distinct mutant morphology in late development. The cellular localization of the protein was elucidated by creating a HelF-GFP fusion protein, which was found to be localized in speckles in the nucleus. The involvement of HelF in the RNAi mechanism was studied. For this purpose, RNAi was induced by transformation of RNAi hairpin constructs against four endogenous genes in wild type and HelF- cells. The silencing efficiency was strongly enhanced in the HelF K.O. strain in comparison with the wild type. One gene, which could not be silenced in the wild type background, was successfully silenced in HelF-. When the helF gene was disrupted in a secondary transformation in a non-silenced strain, the silencing efficiency was strongly improved, a phenomenon named here “retrosilencing”. Transcriptional run-on experiments revealed that the enhanced gene silencing in HelF- was a posttranscriptional event, and that the silencing efficiency depended on the transcription levels of hairpin RNAs. In HelF-, the threshold level of hairpin transcription required for efficient silencing was dramatically lowered. The RNAi-mediated silencing was accompanied by the production of siRNAs; however, their amount did not depend on the level of hairpin transcription. These results indicated that HelF is a natural suppressor of RNAi in Dictyostelium. In contrast, asRNA mediated gene silencing was not enhanced in the HelF K.O, as shown for three tested genes. These results confirmed previous observations (H. Martens and W. Nellen, unpublished) that although similar, RNAi and asRNA mediated gene silencing mechanisms differ in their requirements for specific proteins. In order to characterize the function of the HelF protein on a molecular level and to study its interactions with other RNAi components, in vitro experiments were performed. Besides the DEAH-helicase domain, HelF contains a double-stranded RNA binding domain (dsRBD) at its N-terminus, which showed high similarity to the dsRBD domain of Dicer A from Dictyostelium. The ability of the recombinant dsRBDs from HelF and Dicer A to bind dsRNA was examined and compared. It was shown by gel-shift assays that both HelF-dsRBD and Dicer-dsRBD could bind directly to long dsRNAs. However, HelF-dsRBD bound more efficiently to dsRNA with imperfect matches than to perfect dsRNA. Both dsRBDs bound specifically to a pre-miRNA substrate (pre-let-7). The results suggested that most probably there were two binding sites for the proteins on the pre-miRNA substrate. Moreover, it was shown that HelF-dsRBD and Dicer-dsRBD have siRNA-binding activity. The affinities of the two dsRBDs to the pre-let-7 substrate were also examined by plasmon surface resonance analyses, which revealed a 9-fold higher binding affinity of the Dicer-dsRBD to pre-let-7 compared to that of the HelF-dsRBD. The binding of HelF-dsRBD to the pre-let-7 was impaired in the presence of Mg2+, while the Dicer-dsRBD interaction with pre-let-7 was not influenced by the presence of Mg2+. The results obtained in this thesis can be used to postulate a model for HelF function. In this, HelF acts as a nuclear suppressor of RNAi in wild type cells by recognition and binding of dsRNA substrates. The protein might act as a surveillance system to avoid RNAi initiation by fortuitous dsRNA formation or low abundance of dsRNA trigger. If the protein acts as an RNA helicase, it could unwind fold-back structures in the nucleus and thus lead to decreased RNAi efficiency. A knock-out of HelF would result in initiation of the RNAi pathway even by low levels of dsRNA. The exact molecular function of the protein in the RNAi mechanism still has to be elucidated. RNA interferenz (RNAi) ist ein in jüngster Zeit entdeckter Mechanismus, bei dem doppelsträngige RNA Moleküle (dsRNA) eine Homologie-abhängige Degradation einer verwandten messenger-RNA (mRNA) auslösen. Auf der Suche nach neuen Komponenten der RNAi-Maschinerie in Dictyostelium konnte ein neues Gen (helF) identifiziert werden. HelF ist eine putative RNA-Helikase mit einer hohen Homologie zur Helikasedomäne der bekannten Dicerproteine, der Helikasedomäne der Dictyostelium RdRP und zu dem C. elegans Gen drh-1, welches für eine Dicer-bezogene DExH-box RNA Helikase codiert, die am RNAi-Mechanismus beteiligt ist. Das Ziel dieser Arbeit war es, die Funktion von HelF im Zusammenhang des RNAi oder asRNA induzierten PTGS zu untersuchen. Es wurde eine Unterbrechung des helF-Gens auf genomischer Ebene (K.O.) vorgenommen, was bei den Mutanten zu einer veränderten Morphologie in der späten Entwicklung führte. Die Lokalisation des Proteins in der Zelle konnte mit Hilfe einer GFP-Fusion analysiert werden und kleinen Bereichen innerhalb des Nukleus zugewiesen werden. Im Weiteren wurde der Einfluss von HelF auf den RNAi-Mechanismus untersucht. Zu diesem Zweck wurde RNAi durch Einbringen von RNAi Hairpin-Konstrukten gegen vier endogene Gene im Wiltypstamm und der HelF--Mutante induziert. Im Vergleich zum Wildtypstamm konnte im HelF--Mutantenstamm eine stark erhöhte „Silencing“-Effizienz nachgewiesen werden. Ein Gen, welches nach RNAi Initiation im Wildtypstamm unverändert blieb, konnte im HelF--Mutantenstamm erfolgreich stillgelegt werden. Durch sekundäres Einführen einer Gendisruption im helF-Locus in einen Stamm, in welchem ein Gen nicht stillgelegt werden konnte, wurde die Effizienz des Stilllegens deutlich erhöht. Dieses Phänomen wurde hier erstmals als „Retrosilencing“ beschrieben. Mit Hilfe von transkriptionellen run-on Experimenten konnte belegt werden, dass es sich bei dieser erhöhten Stilllegungseffizienz um ein posttranskriptionelles Ereignis handelte, wobei die Stillegungseffizienz von der Transkriptionsstärke der Hairpin RNAs abhängt. Für die HelF--Mutanten konnte gezeigt werden, dass der Schwellenwert zum Auslösen eines effizienten Stillegens dramatisch abgesenkt war. Obwohl die RNAi-vermittelte Genstilllegung immer mit der Produktion von siRNAs einhergeht, war die Menge der siRNAs nicht abhängig von dem Expressionsniveau des Hairpin-Konstruktes. Diese Ergebnisse legen nahe, dass es sich bei der HelF um einen natürlichen Suppressor des RNAi-Mechanismus in Dictyostelium handelt. Im Gegensatz hierzu war die as-vermittelte Stilllegung von drei untersuchten Genen im HelF-K.O. im Vergleich zum Wildyp unverändert. Diese Ergebnisse bestätigten frühere Beobachtungen (H. Martens und W. Nellen, unveröffentlicht), wonach die Mechanismen für RNAi und asRNA-vermittelte Genstilllegung unterschiedliche spezifische Proteine benötigen. Um die Funktion des HelF-Proteins auf der molekularen Ebene genauer zu charakterisieren und die Interaktion mit anderen RNAi-Komponenten zu untersuchen, wurden in vitro Versuche durchgeführt. Das HelF-Protein enthält, neben der DEAH-Helikase-Domäne eine N-terminale Doppelstrang RNA bindende Domäne (dsRBD) mit einer hohen Ähnlichkeit zu der dsRBD des Dicer A aus Dictyostelium. Die dsRNA-Bindungsaktivität der beiden dsRBDs aus HelF und Dicer A wurde analysiert und verglichen. Es konnte mithilfe von Gel-Retardationsanalysen gezeigt werden, dass sowohl HelF-dsRBD als auch Dicer-dsRBD direkt an lange dsRNAs binden können. Hierbei zeigte sich, dass die HelF-dsRBD eine höhere Affinität zu einem imperfekten RNA-Doppelstrang besitzt, als zu einer perfekt gepaarten dsRNA. Für beide dsRBDs konnte eine spezifische Bindung an ein pre-miRNA Substrat nachgewiesen werden (pre-let-7). Dieses Ergebnis legt nah, dass es zwei Bindestellen für die Proteine auf dem pre-miRNA Substrat gibt. Überdies hinaus konnte gezeigt werden, dass die dsRBDs beider Proteine eine siRNA bindende Aktivität besitzen. Die Affinität beider dsRBDs an das pre-let-7 Substrat wurde weiterhin mit Hilfe der Plasmon Oberflächen Resonanz untersucht. Hierbei konnte eine 9-fach höhere Bindeaffinität der Dicer-dsRBD im Vergleich zur HelF-dsRBD nachgewiesen werden. Während die Bindung der HelF-dsRBD an das pre-let-7 durch die Anwesenheit von Mg2+ beeinträchtigt war, zeigte sich kein Einfluß von Mg2+ auf das Bindeverhalten der Dicer-dsRBD. Mit Hilfe der in dieser Arbeit gewonnen Ergebnisse lässt sich ein Model für die Funktion von HelF postulieren. In diesem Model wirkt HelF durch Erkennen und Binden von dsRNA Substraten als Suppressor von der RNAi im Kern. Das Protein kann als Überwachungsystem gegen eine irrtümliche Auslösung von RNAi wirken, die durch zufällige dsRNA Faltungen oder eine zu geringe Häufigkeit der siRNAs hervorgerufen sein könnte. Falls das Protein eine Helikase-Aktivität besitzt, könnte es rückgefaltete RNA Strukturen im Kern auflösen, was sich in einer verringerten RNAi-Effizienz wiederspiegelt. Durch Ausschalten des helF-Gens würde nach diesem Modell eine erfolgreiche Auslösung von RNAi schon bei sehr geringer Mengen an dsRNA möglich werden. Das Modell erlaubt, die exakte molekulare Funktion des HelF-Proteins im RNAi-Mechanismus weiter zu untersuchen.

Untersuchung der Sensitivität von Heizungs- / Lüftungsanlagen in Niedrigenergiehäusern

Bei Meßprojekten in denen Gebäude mit installierter Lüftungsanlage untersucht wurden, stellte man immer wieder ein breite Streuung der Meßwerte, als auch eine oftmals deutliche Abweichung vom vorher ermittelten Heizwärmebedarf der Gebäude fest. Es wird vermutet, daß diese Unterschiede systemspezifische Ursachen haben, ein Nachweis kann aufgrund der geringen Anzahl vorhandener Meßpunkte jedoch nicht geführt werden. Um die Sensitivität verschiedener Randbedingungen auf den Energieverbrauch zu ermitteln, wird im vorliegenden Forschungsprojekt ein Simulationsmodell erstellt. Das thermische Verhalten und die Durchströmung des Gebäudes werden durch ein gekoppeltes Modell abgebildet. Unterschiedliche Lüftungsanlagensysteme werden miteinander verglichen. Auf Basis vorhandener Meßdaten wird ein klimaabhängiges Modell zur Fensterlüftung entwickelt, welches in die Modellbildung der Gebäudedurchströmung mit einfließt. Feuchtegeregelte Abluftanlagen sind in der Lage den mittleren Luftwechsel auf ein hygienisch sinnvollen Wert zu begrenzen. Sie erweisen sich im Hinblick auf die Sensitivität verschiedener Randbedingungen als robuste Systeme. Trotz Einsatz von Lüftungsanlagen kann je nach Betriebszustand insbesondere bei Abluftanlagen keine ausreichende Luftqualität sichergestellt werden. Zukünftige Systeme dürfen das "Lüftungssystem" Fenster nicht vernachlässigen, sondern müssen es in das Gesamtkonzept mit einbeziehen.

Die Auswirkungen von Hochgeschwindigkeitsverkehr auf die Erreichbarkeit der Regionen in Deutschland - dargestellt am Beispiel der Magnetschwebebahn Transrapid

Nach 35 Jahren Entwicklungszeit wurde im Jahr 2004 in Shanghai die erste kommerzielle Anwendung des innovativen Magnetbahnsystems Transrapid in Betrieb genommen; in Deutschland konnte bislang keine Transrapid-Strecke realisiert werden, obwohl dieses System entsprechend den Ergebnissen einer vom damaligen Bundesverkehrsminister beauftragten Studie aus dem Jahr 1972 für den Einsatz in Deutschland entwickelt wurde. Beim Transrapid handelt es sich um eine echte Produkt-Innovation im Bahnverkehr und nicht um eine Weiterentwicklung oder Optimierung wie beim ICE, und ist somit als innovativer Verkehrsträger der Zukunft in die langfristige Entwicklung der Verkehrssysteme einzufügen. Die modernen HGV Bahnsysteme (Shinkansen/TGV/ICE) hingegen sind, ähnlich der Clipper in der Phase der Segelschifffahrt im Übergang zum Dampfschiff, letzte Abwehrentwicklungen eines am Zenit angekommenen Schienen-Verkehrssystems. Die Einführung von Innovationen in einen geschlossenen Markt stellt sich als schwierig dar, da sie zu einem Bruch innerhalb eines etablierten Systems führen. Somit wird in der vorliegenden Arbeit im ersten Teil der Themenkomplex Innovation und die Einordnung der Magnet-Schwebe-Technologie in diese langfristig strukturierten Abläufe untersucht und dargestellt. Das Transrapid-Projekt ist demzufolge in eine zeitstrukturelle Zyklizität einzuordnen, die dafür spricht, die Realisierung des Gesamtprojektes in eine Zeitspanne von 20 bis 30 Jahre zu verlagern. Im zweiten Teil wird auf der Basis einer regionalstrukturellen Analyse der Bundesrepublik Deutschland ein mögliches Transrapidnetz entworfen und die in diesem Netz möglichen Reisezeiten simuliert. Weiterhin werden die Veränderungen in den Erreichbarkeiten der einzelnen Regionen aufgrund ihrer Erschließung durch das Transrapidnetz simuliert und grafisch dargestellt. Die vorliegende Analyse der zeitlichen Feinstruktur eines perspektiven Transrapidnetzes ist ein modellhafter Orientierungsrahmen für die Objektivierung von Zeitvorteilen einer abgestimmten Infrastruktur im Vergleich zu real möglichen Reisezeiten innerhalb Deutschlands mit den gegebenen Verkehrsträgern Schiene, Straße, Luft. So würde der Einsatz des Transrapid auf einem entsprechenden eigenständigen Netz die dezentrale Konzentration von Agglomerationen in Deutschland fördern und im Durchschnitt annähernd 1 h kürzere Reisezeiten als mit den aktuellen Verkehrsträgern ermöglichen. Zusätzlich wird noch ein Ausblick über mögliche Realisierungsschritte eines Gesamtnetzes gegeben und die aufgetretenen Schwierigkeiten bei der Einführung des innovativen Verkehrssystems Transrapid dargestellt.

Composition and stability of organic matter fractions in soils under different land use regimes

Soil organic matter (SOM) vitally impacts all soil functions and plays a key role in the global carbon (C) cycle. More than 70% of the terrestric C stocks that participate in the active C cycle are stored in the soil. Therefore, quantitative knowledge of the rates of C incorporation into SOM fractions of different residence time is crucial to understand and predict the sequestration and stabilization of soil organic carbon (SOC). Consequently, there is a need of fractionation procedures that are capable of isolating functionally SOM fractions, i.e. fractions that are defined by their stability. The literature generally refers to three main mechanisms of SOM stabilization: protection of SOM from decomposition by (i) its structural composition, i.e. recalcitrance, (ii) spatial inaccessibility and/or (iii) interaction with soil minerals and metal ions. One of the difficulties in developing fractionation procedures for the isolation of functional SOM fractions is the marked heterogeneity of the soil environment with its various stabilization mechanisms – often several mechanisms operating simultaneously – in soils and soil horizons of different texture and mineralogy. The overall objective of the present thesis was to evaluate present fractionation techniques and to get a better understanding of the factors of SOM sequestration and stabilization. The first part of this study is attended to the structural composition of SOM. Using 13C cross-polarization magic-angle spinning (CPMAS) nuclear magnetic resonance (NMR) spectroscopy, (i) the effect of land use on SOM composition was investigated and (ii) examined whether SOM composition contributes to the different stability of SOM in density and aggregate fractions. The second part of the present work deals with the mineral-associated SOM fraction. The aim was (iii) to evaluate the suitability of chemical fractionation procedures used in the literature for the isolation of stable SOM pools (stepwise hydrolysis, treatments using oxidizing agents like Na2S2O8, H2O2, and NaOCl as well as demineralization of the residue obtained by the NaOCl treatment using HF (NaOCl+HF)) by pool sizes, 13C and 14C data. Further, (iv) the isolated SOM fractions were compared to the inert organic matter (IOM) pool obtained for the investigated soils using the Rothamsted Carbon Model and isotope data in order to see whether the tested chemical fractionation methods produce SOM fractions capable to represent this pool. Besides chemical fractionation, (v) the suitability of thermal oxidation at different temperatures for obtaining stable SOC pools was evaluated. Finally, (vi) the short-term aggregate dynamics and the factors that impact macroaggregate formation and C stabilization were investigated by means of an incubation study using treatments with and without application of 15N labeled maize straw of different degradability (leaves and coarse roots). All treatments were conducted with and without the addition of fungicide. Two study sites with different soil properties and land managements were chosen for these investigations. The first one, located at Rotthalmünster, is a Stagnic Luvisol (silty loam) under different land use regimes. The Ah horizons of a spruce forest and continuous grassland and the Ap and E horizons of two plots with arable crops (continuous maize and wheat cropping) were examined. The soil of the second study site, located at Halle, is a Haplic Phaeozem (loamy sand) where the Ap horizons of two plots with arable crops (continuous maize and rye cropping) were investigated. Both study sites had a C3-/C4-vegetational change on the maize plot for the purpose of tracing the incorporation of the younger, maize-derived C into different SOM fractions and the calculation of apparent C turnover times of these. The Halle site is located near a train station and industrial areas, which caused a contamination with high amounts of fossil C. The investigation of aggregate and density fractions by 13C CPMAS NMR spectroscopy revealed that density fractionation isolated SOM fractions of different composition. The consumption of a considerable part (10–20%) of the easily available O-alkyl-C and the selective preservation of the more recalcitrant alkyl-C when passing from litter to the different particulate organic matter (POM) fractions suggest that density fractionation was able to isolate SOM fractions with different degrees of decomposition. The spectra of the aggregate fractions resembled those of the mineral-associated SOM fraction obtained by density fractionation and no considerable differences were observed between aggregate size classes. Comparison of plant litter, density and aggregate size fractions from soil under different land use showed that the type of land use markedly influenced the composition of SOM. While SOM of the acid forest soil was characterized by a large content (> 50%) of POM, which contained high amounts of spruce-litter derived alkyl-C, the organic matter in the biologically more active grassland and arable soils was dominated by mineral-associated SOM (> 95%). This SOM fraction comprised greater proportions of aryl- and carbonyl-C and is considered to contain a higher amount of microbially-derived organic substances. Land use can alter both, structure and stability of SOM fractions. All applied chemical treatments induced considerable SOC losses (> 70–95% of mineral-associated SOM) in the investigated soils. The proportion of residual C after chemical fractionation was largest in the arable Ap and E horizons and increased with decreasing C content in the initial SOC after stepwise hydrolysis as well as after the oxidative treatments with H2O2 and Na2S2O8. This can be expected for a functional stable pool of SOM, because it is assumed that the more easily available part of SOC is consumed first if C inputs decrease. All chemical treatments led to a preferential loss of the younger, maize-derived SOC, but this was most pronounced after the treatments with Na2S2O8 and H2O2. After all chemical fractionations, the mean 14C ages of SOC were higher than in the mineral-associated SOM fraction for both study sites and increased in the order: NaOCl < NaOCl+HF ≤ stepwise hydrolysis << H2O2 ≈ Na2S2O8. The results suggest that all treatments were capable of isolating a more stable SOM fraction, but the treatments with H2O2 and Na2S2O8 were the most efficient ones. However, none of the chemical fractionation methods was able to fit the IOM pool calculated using the Rothamsted Carbon Model and isotope data. In the evaluation of thermal oxidation for obtaining stable C fractions, SOC losses increased with temperature from 24–48% (200°C) to 100% (500°C). In the Halle maize Ap horizon, losses of the young, maize-derived C were considerably higher than losses of the older C3-derived C, leading to an increase in the apparent C turnover time from 220 years in mineral-associated SOC to 1158 years after thermal oxidation at 300°C. Most likely, the preferential loss of maize-derived C in the Halle soil was caused by the presence of the high amounts of fossil C mentioned above, which make up a relatively large thermally stable C3-C pool in this soil. This agrees with lower overall SOC losses for the Halle Ap horizon compared to the Rotthalmünster Ap horizon. In the Rotthalmünster soil only slightly more maize-derived than C3-derived SOC was removed by thermal oxidation. Apparent C turnover times increased slightly from 58 years in mineral-associated SOC to 77 years after thermal oxidation at 300°C in the Rotthalmünster Ap and from 151 to 247 years in the Rotthalmünster E horizon. This led to the conclusion that thermal oxidation of SOM was not capable of isolating SOM fractions of considerably higher stability. The incubation experiment showed that macroaggregates develop rapidly after the addition of easily available plant residues. Within the first four weeks of incubation, the maximum aggregation was reached in all treatments without addition of fungicide. The formation of water-stable macroaggregates was related to the size of the microbial biomass pool and its activity. Furthermore, fungi were found to be crucial for the development of soil macroaggregates as the formation of water-stable macroaggregates was significantly delayed in the fungicide treated soils. The C concentration in the obtained aggregate fractions decreased with decreasing aggregate size class, which is in line with the aggregate hierarchy postulated by several authors for soils with SOM as the major binding agent. Macroaggregation involved incorporation of large amounts maize-derived organic matter, but macroaggregates did not play the most important role in the stabilization of maize-derived SOM, because of their relatively low amount (less than 10% of the soil mass). Furthermore, the maize-derived organic matter was quickly incorporated into all aggregate size classes. The microaggregate fraction stored the largest quantities of maize-derived C and N – up to 70% of the residual maize-C and -N were stored in this fraction.

Numerical Feasibility Study for Treated Wastewater Recharge as a Tool to Impede Saltwater Intrusion in the Coastal Aquifer of Gaza – Palestine

The ongoing depletion of the coastal aquifer in the Gaza strip due to groundwater overexploitation has led to the process of seawater intrusion, which is continually becoming a serious problem in Gaza, as the seawater has further invaded into many sections along the coastal shoreline. As a first step to get a hold on the problem, the artificial neural network (ANN)-model has been applied as a new approach and an attractive tool to study and predict groundwater levels without applying physically based hydrologic parameters, and also for the purpose to improve the understanding of complex groundwater systems and which is able to show the effects of hydrologic, meteorological and anthropogenic impacts on the groundwater conditions. Prediction of the future behaviour of the seawater intrusion process in the Gaza aquifer is thus of crucial importance to safeguard the already scarce groundwater resources in the region. In this study the coupled three-dimensional groundwater flow and density-dependent solute transport model SEAWAT, as implemented in Visual MODFLOW, is applied to the Gaza coastal aquifer system to simulate the location and the dynamics of the saltwater–freshwater interface in the aquifer in the time period 2000-2010. A very good agreement between simulated and observed TDS salinities with a correlation coefficient of 0.902 and 0.883 for both steady-state and transient calibration is obtained. After successful calibration of the solute transport model, simulation of future management scenarios for the Gaza aquifer have been carried out, in order to get a more comprehensive view of the effects of the artificial recharge planned in the Gaza strip for some time on forestall, or even to remedy, the presently existing adverse aquifer conditions, namely, low groundwater heads and high salinity by the end of the target simulation period, year 2040. To that avail, numerous management scenarios schemes are examined to maintain the ground water system and to control the salinity distributions within the target period 2011-2040. In the first, pessimistic scenario, it is assumed that pumping from the aquifer continues to increase in the near future to meet the rising water demand, and that there is not further recharge to the aquifer than what is provided by natural precipitation. The second, optimistic scenario assumes that treated surficial wastewater can be used as a source of additional artificial recharge to the aquifer which, in principle, should not only lead to an increased sustainable yield of the latter, but could, in the best of all cases, revert even some of the adverse present-day conditions in the aquifer, i.e., seawater intrusion. This scenario has been done with three different cases which differ by the locations and the extensions of the injection-fields for the treated wastewater. The results obtained with the first (do-nothing) scenario indicate that there will be ongoing negative impacts on the aquifer, such as a higher propensity for strong seawater intrusion into the Gaza aquifer. This scenario illustrates that, compared with 2010 situation of the baseline model, at the end of simulation period, year 2040, the amount of saltwater intrusion into the coastal aquifer will be increased by about 35 %, whereas the salinity will be increased by 34 %. In contrast, all three cases of the second (artificial recharge) scenario group can partly revert the present seawater intrusion. From the water budget point of view, compared with the first (do nothing) scenario, for year 2040, the water added to the aquifer by artificial recharge will reduces the amount of water entering the aquifer by seawater intrusion by 81, 77and 72 %, for the three recharge cases, respectively. Meanwhile, the salinity in the Gaza aquifer will be decreased by 15, 32 and 26% for the three cases, respectively.