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Effects of Natural/anthropogenic Stressors and a Chemical Contaminant on Pre and Post Mycorrhizal Colonization in Wetland Plants

Description: Arbuscular mycorrhizal fungi, colonizing over 80% of all plants, were long thought absent in wetlands; however, recent studies have shown many wetland plants harbor arbuscular mycorrhizae (AM) and dark septate endophytes (DSE). Wetland services such as biodiversity, shoreline stabilization, water purification, flood control, etc. have been estimated to have a global value of $14.9 trillion. Recognition of these vital services is accompanied by growing concern for their vulnerability and continued loss, which has resulted in an increased need to understand wetland plant communities and mycorrhizal symbiosis. Factors regulating AM and DSE colonization need to be better understood to predict plant community response and ultimately wetland functioning when confronting natural and human induced stressors. This study focused on the effects of water quality, hydrology, sedimentation, and hurricanes on AM and DSE colonization in three wetland species (Taxodium distichum, Panicum hemitomon, and Typhal domingensis) and plant communities of coastal wetlands in Southeast Louisiana and effects of an antimicrobial biocide, triclosan (TCS), on AM (Glomus intraradices) spore germination, hyphal growth, hyphal branching, and colonization in fresh water wetland plants (Eclipta prostrata, Hibiscus laevis, and Sesbania herbacea) from bottom land hardwood forest in north central Texas. The former, mesocosm studies simulating coastal marsh vegetation ran for five years. In the latter studies, AM spores and wetland plants were exposed to 0 g/L, 0.4 g/L, and 4.0 g/L TCS concentrations in static renewal and flow through exposures for 21 and 30 days, respectively. AM and DSE colonization was significantly affected by individual and interactions of four independent variables in mesocosm experiments. Similarly, spore germination, hyphal growth, hyphal branching, and AM colonization in selected wetland plants were significantly lowered by exposure to the TCS at environmentally relevant concentrations. However, levels of effects were plant species and fungal propagules specific. My results showed that natural and human ...
Date: August 2013
Creator: Twanabasu, Bishnu Ram
Partner: UNT Libraries

Development of Enabling Technologies to Visualize the Plant Lipidome

Description: Improvements in mass spectrometry (MS)-based strategies for characterizing the plant lipidome through quantitative and qualitative approaches such as shotgun lipidomics have substantially enhanced our understanding of the structural diversity and functional complexity of plant lipids. However, most of these approaches require chemical extractions that result in the loss of the original spatial context and cellular compartmentation for these compounds. To address this current limitation, several technologies were developed to visualize lipids in situ with detailed chemical information. A subcellular visualization approach, direct organelle MS, was developed for directly sampling and analyzing the triacylglycerol contents within purified lipid droplets (LDs) at the level of a single LD. Sampling of single LDs demonstrated seed lipid droplet-to-droplet variability in triacylglycerol (TAG) composition suggesting that there may be substantial variation in the intracellular packaging process for neutral lipids in plant tissues. A cellular and tissue visualization approach, MS imaging, was implemented and enhanced for visualizing the lipid distributions in oilseeds. In mature cotton seed embryos distributions of storage lipids (TAGs) and their phosphatidylcholine (PCs) precursors were distribution heterogeneous between the cotyledons and embryonic axis raising new questions about extent and regulation of oilseed heterogeneity. Extension of this methodology provides an avenue for understanding metabolism in cellular (perhaps even subcellular) context with substantial metabolic engineering implications. To visualize metabolite distributions, a free and customizable application, Metabolite Imager, was developed providing several tools for spatially-based chemical data analysis. These tools collectively enable new forms of visualizing the plant lipidome and should prove valuable toward addressing additional unanswered biological questions.
Date: August 2013
Creator: Horn, Patrick J.
Partner: UNT Libraries

Endocrine Disruption of Levonorgestrel in Early-life Stages of Fathead Minnows, Pimephales Promelas

Description: Pharmaceuticals have routinely been detected in the environment resulting in a growing concern about whether these drugs could elicit effects on aquatic organisms. The concerns are centered on the highly conserved nature of mammalian therapeutic targets in fish. These pharmaceuticals are found at very low levels in the environment, which can result in sub-lethal effects in aquatic organisms. Therefore, 28 d early-life stage studies were conducted on six pharmaceuticals to assess their impacts on survival and growth fathead minnow larvae. Two pharmaceuticals tested, carbamazepine and fenofibrate, resulted in no alterations to survival and growth. However, amiodarone, clozapine, dexamethasone, and levonorgestrel (LNG) reduced survival at concentrations tested with LNG being the most potent at 462 ng/L. Survival was increased with amiodarone and clozapine; however LNG significantly decreased growth at 86 ng/L. Therefore, the most potent pharmaceutical tested was the synthetic progestin LNG with survival and growth impacts at concentrations less than 1 μg/L. Further analysis was conducted by measuring specific endocrine related mRNA transcript profiles in FHM larvae following the 28 d ELS exposure to LNG. Transcripts of 3β-HSD, 20β-HSD, and FSH were significantly down-regulated following 28 d exposure to both 16.3 and 86.9 ng/L LNG. Also, CYP19a expression was significantly down-regulated at 86.9 and 2392 ng/L LNG. Subsequently, a second study examined time periods that may be most sensitive (e.g., windows of sensitivity) for FHM larvae exposed to LNG. Larvae were exposed to a single concentration of LNG (i.e. LOECgrowth of 86.2 ng/L as determined in the 28 d ELS study) for different time periods starting with fertilized egg through 28 dph. Growth and mRNA expression of the four differentially expressed transcripts from the first study were measured. Regardless of the duration of exposure, LNG significantly decreased growth in fathead minnow larvae at day 28. For both 20β-HSD and CYP19a, ...
Date: August 2013
Creator: Overturf, Matthew D.
Partner: UNT Libraries

Aging Is a Determinant in Anoxia Stress Tolerance in Caenorhabditis Elegans

Description: Oxygen availability is critical for survival for most organisms. The nematode, C. elegans, has been useful for studying genetic regulation of anoxia tolerance due to the oxygen deprivation response mechanisms shared with other metazoans. Studies examining long-term anoxia (72h, LTA) tolerance have only been conducted at adult day 1. To investigate the effect of aging on anoxia tolerance wild-type and mutant strains were exposed to LTA between adult day 1 and day 9. Wild-type isolates and daf-16(mu86) (FOXO transcription factor regulated by insulin-signaling) and aak-2(gt33) (catalytic subunit of AMP-activated protein kinase) strains were anoxia sensitive at day 1 and displayed increased LTA tolerance with aging correlated with reproductive senescence followed by a decline in survivorhsip through day 9. The daf-2(e1370) (insulin receptor homologue of C. elegans), glp-1(e2141) (a lin-12/Notch receptor) and fog-2(q71) (required for spermatogenesis) strains were LTA-tolerant through day 5. I conclude that aging influences LTA-tolerance in a strain- and age-dependent manner. In addition to being LTA-tolerant the daf-2(e1370) and glp-1(e2141) strains have a longevity phenotype that is suppressed by loss of kri-1 or daf-12. While loss of kri-1 did not suppress the LTA-tolerant phenotype of glp-1(e2141) at day 1 the portion of impaired survivors increased at day 3 and by day 5 tolerance was suppressed. Similarly, when exposed to 4 days of anoxia the glp-1(e2141);daf-12(rh41rh611) double mutant had a reduced survivor rate at all ages analyzed compared to glp-1(e2141) controls. To better understand formation of an anoxia-tolerant physiology I exposed adults to one or more 24h bouts. Recurrent bouts increased LTA tolerance in wild-type hermaphrodites in a dose-dependent manner. Bout-treated daf-16(mu86) animals had increased survival rate compared to controls yet maximum survival remained below age-matched wild-type. Anoxia bouts decreased LTA-tolerance in aak-2(gt33) mutants, indicating the requirement for ATP regulation in establishing an LTA-tolerant phenotype. These data support the ...
Date: May 2013
Creator: Goy, Jo M.
Partner: UNT Libraries

Field and Laboratory Fish Tissue Accumulation of Carbamazepine and Amiodarone

Description: The goals of this dissertation work were to assess the bioaccumulation potential of carbamazepine and amiodarone, two widely used ionizable pharmaceutical compounds that possess mid-range and high LogD values, respectively, and to evaluate alternative methods to assess chemical accumulation in bluntnose minnows, catfish, and tilapia. Results indicated that carbamazepine does not appreciably bioaccumulate in fish tissue with BCFk and BAF carbamazepine values < 10. Amiodarone, however, with a log D of 5.87 at pH 7.4, accumulated in fish tissues with kinetic BCF values <2,400. Collectively, the data suggest that full and abbreviated laboratory-derived BCFs, BCFMs derived from S9 loss-of-parent assays, as well as field BAF values are similar for each of the two drugs. In summary, the results from this dissertation indicated: 1) The reduced design BCF test is a good estimate for the traditional OECD 305 test. 2) In vitro S9 metabolism assays provide comparable BCF estimates to the OECD 305 test. 3) Metabolism may play a large role in the accumulation of drugs in fish. 4) Reduced BCF tests and in vitro assays are cost effective and can reduce vertebrate testing.
Date: December 2013
Creator: García Martínez, Santos Noé
Partner: UNT Libraries

Population Dynamics of Zebra Mussels (Dreissena Polymorpha) in a North Texas Reservoir: Implications for Invasions in the Southern United States

Description: This dissertation has two main objectives: first, quantify the effects of environmental conditions on spatio-temporal spawning and larval dynamics of zebra mussels (Dreissena polymorpha [Pallas 1771]) in Lake Texoma, and second, quantify the effects of environmental conditions on survival, growth, and reproduction of young of the year (YOY) juvenile zebra mussels. These biological responses directly influence population establishment success and invasive spread dynamics. Reproductive output of the zebra mussel population in Lake Texoma was significantly related to water temperature and lake elevation. Annual maximum larval (veliger) density decreased significantly indicating a population crash, which was likely caused by thermal stress and variability of lake elevation. In 2011, temperatures peaked at 34.3°C and lake elevation decreased to the lowest level recorded during the previous 18 years, which desiccated a substantial number of settled mussels in littoral zones. Estimated mean date of first spawn in Lake Texoma was observed approximately 1.5 months earlier than in Lake Erie, and peak veliger densities were observed two months earlier. Veligers were observed in the deepest oxygenated water after lake stratification. During a 69-day in situ experiment during summer in Lake Texoma, age-specific mortality of zebra mussels was generally high until temperatures decreased to approximately 28°C, which was observed after lake turnover in late summer. No study organism died after temperatures decreased to less than 26°C, which indicates individuals that survive high summer temperatures are likely to persist into autumn/winter. Shell length growth and soft tissue growth rates were related to temperature and chlorophyll-a concentration, respectively. Growth rates of study organisms were among the highest ever reported for D. polymorpha. Water temperature and body size influenced reproduction of YOY zebra mussels in Lake Texoma. Fecundity of females were positively related to temperature; however, sperm production was negatively related to temperature, which indicates males could be more ...
Date: December 2013
Creator: Churchill, Christopher J.
Partner: UNT Libraries

Modeling the Effects of Chronic Toxicity of Pharmaceutical Chemicals on the Life History Strategies of Ceriodaphnia Dubia: a Multigenerational Study

Description: Trace quantities of pharmaceuticals (including carbamazepine and sertraline) are continuously discharged into the environment, which causes concern among scientists and regulators regarding their potential long-term impacts on aquatic ecosystems. These compounds and their metabolites are continuously interacting with the orgranisms in various life stages, and may differentially influence development of embryo, larvae, juvenile, and adult stages. To fully understand the potential ecological risks of two candidate pharmaceutical chemicals (carbamazepine (CBZ) and sertraline (SERT)) exposure on survival, growth and reproduction of Ceriodaphnia dubia in three sucessive generations under static renewal toxicity test, a multigenerational approach was taken. Results indicate that SERT exposure showed higher sensitivity to chronic exposure to C. dubia growth and reproduction than CBZ exposure. The lowest concentration to affect fecundity and growth was at 50 µg L-1 SERT in the first two generations. These parameters become more sensitive during the third generation where the LOEC was 4.8 µg L-1. The effective concentrations (EC50) for the number of offspring per female, offspring body size, and dry weight were 17.2, 21.2, and 26.2 µg SERT L-1, respectively. Endpoints measured in this study demonstrate that chronic exposure of C. dubia to SERT leads to effects that occur at concentrations an order of magnitude higher than predicted environmental concentrations indicating potential transgenerationals effects. Additionally, a process-based dynamic energy budget (DEB) model is implemented to predict the simulated effects of chronic toxicity of SERT and CBZ to C. dubia individual behavior at laboratory condition. The model‘s output indicates the ecotoxicological mode of action of SERT exposure, which acts on feeding or assimilation with an effect that rapidly saturates at higher concentrations. Offspring size decreases with the toxic effects on feeding, and offspring number is thus less affected than total investment in reproduction. Consequently, CBZ affects direclty in reproduction which are captured by DEBtox ...
Date: December 2013
Creator: Lamichhane, Kiran
Partner: UNT Libraries

Relationships of Benthic Macroinvertebrate Community Structure with Land-use, Habitat, In-stream Water Chemistry, Depositional Sediment Biofilm Fatty Acids, and Surfactants in the Effluent Dominated Texas Trinity River

Description: The Trinity River is an urbanized, effluent-dominated river, and is heavily relied upon for drinking water. The benthic macroinvertebrate community has been monitored for over 20 years, with the focus of this dissertation on three studies (1987-88, 2005, and 2011). Water quality improvement following dechlorination resulted in increased benthic metrics. Overall habitat quality, in-stream cover, surface water total organic carbon, sediment total organic carbon, near-field urban land-use, near-field forested land-use, surface water surfactant toxic units, and depositional sediment biofilm fatty acids all have statistically significant relationships with benthic macroinvertebrate metrics. These relationships are better defined with increased taxonomic resolution at the genus/species level for all benthic taxa, including Chironomidae and Oligochaeta. It is recommend that benthic identifications for state and city water quality assessments be done at the genus/species level. A novel method for quantifying depositional sediment biofilm fatty acids has been produced and tested in this dissertation. Benthic metrics are directly related to fatty acid profiles, with several essential fatty acids found only at upstream sites.
Date: December 2013
Creator: Slye, Jaime L.
Partner: UNT Libraries

Investigation of the Pharmacokinetics of Diazepam in Juvenile Channel Catfish (Ictalurus Punctatus)

Description: The presence of pharmaceuticals in the environment is becoming an increasing regulatory and scientific concern. Thus, the metabolic profile and bioconcentration potential of diazepam, a model benzodiazepine, were examined, as well as effects on the endocrine system in channel catfish. Through the use of specific and non-specific cytochrome P450 (CYP450) inhibitors, it was determined that CYP3A-like enzymes may play a role in the biotransformation of diazepam into temazepam; however, the isoform(s) required for the formation of other metabolites is still unknown. Overall, only around 7-8% of diazepam is biotransformed into two known metabolites. Due to the lack of inherent metabolism of diazepam in channel catfish, further analysis was conducted to determine the tissue-specific bioconcentration potential of diazepam in catfish. Various tissues were analyzed for the presence of diazepam as well as metabolites and bioconcentration factors (BCF) were calculated, which were all well below regulatory threshold values (> 2000). Additionally, modulation of the endocrine system by diazepam was examined by measuring steroid hormone concentrations and analyzing mRNA expression of selected steroidogenic enzymes and receptors. Two steroidogenic enzymes were modulated following diazepam exposure, indicating potential endocrine disrupting properties of diazepam. Together, these data suggest that diazepam exhibits low metabolic transformation rates in channel catfish, which may lead to accumulation of benzodiazepine compounds that may negatively affect the endocrine system. However, further studies should be aimed at identifying other steroidogenic enzymes and/or receptors that may be modulated following diazepam exposure.
Date: December 2013
Creator: Overturf, Carmen L.
Partner: UNT Libraries

Quantifying Forest Vertical Structure to Determine Bird Habitat Quality in the Greenbelt Corridor, Denton, Tx

Description: This study presents the integration of light detection and range (LiDAR) and hyperspectral remote sensing to create a three-dimensional bird habitat map in the Greenbelt Corridor of the Elm Fork of the Trinity River. This map permits to examine the relationship between forest stand structure, landscape heterogeneity, and bird community composition. A biannual bird census was conducted at this site during the breeding seasons of 2009 and 2010. Census data combined with the three-dimensional map suggest that local breeding bird abundance, community structure, and spatial distribution patterns are highly influenced by vertical heterogeneity of vegetation surface. For local breeding birds, vertical heterogeneity of canopy surface within stands, connectivity to adjacent forest patches, largest forest patch index, and habitat (vegetation) types proved to be the most influential factors to determine bird community assemblages. Results also highlight the critical role of secondary forests to increase functional connectivity of forest patches. Overall, three-dimensional habitat descriptions derived from integrated LiDAR and hyperspectral data serve as a powerful bird conservation tool that shows how the distribution of bird species relates to forest composition and structure at various scales.
Date: August 2013
Creator: Matsubayashi, Shiho
Partner: UNT Libraries

Manipulating Sucrose Proton Symporters to Understand Phloem Loading

Description: Phloem vascular tissues transport sugars synthesized by photosynthesis in mature leaves by a process called phloem loading in source tissues and unloading in sink tissues. Phloem loading in source leaves is catalyzed by Suc/H+ symporters (SUTs) which are energized by proton motive force. In Arabidopsis the principal and perhaps exclusive SUT catalyzing phloem loading is AtSUC2. In mutant plants harboring a T-DNA insertion in each of the functional SUT-family members, only Atsuc2 mutants demonstrate overtly debilitated phloem transport. Analysis of a mutant allele (Atsuc2-4) of AtSUC2 with a T-DNA insertion in the second intron showed severely stunted phenotype similar to previously analyzed Atsuc2 null alleles. However unlike previous alleles Atsuc2-4 produced viable seeds. Analysis of phloem specific promoters showed that promoter expression was regulated by Suc concentration. Unlike AtSUC2p, heterologous promoter CoYMVp was not repressed under high Suc conc. Further analysis was conducted using CoYMVp to test the capacity of diverse clades in SUT-gene family for transferring Suc in planta in Atsuc2 - / - mutant background. AtSUC1 and ZmSUT1 from maize complemented Atsuc2 mutant plants to the highest level compared to all other transporters. Over-expression of the above SUTs in phloem showed enhanced Suc loading and transport, but against expectations, plants were stunted. The implications of SUT over-expression to enhance phloem transport and loading are discussed and how it induces a perception of phosphate imbalance is presented.
Date: August 2013
Creator: Dasgupta, Kasturi
Partner: UNT Libraries

Expression of G-protein Coupled Receptors in Young and Mature Thrombocytes and Knockdown of Gpr18 in Zebrafish

Description: In this study, a novel method based on biotinylated antibodies and streptavidin coated magnetic beads was used to separate the thrombocyte subpopulations from zebrafish whole blood. DiI-C18, a lipophilic dye, labels only young thrombocytes when used at low concentrations. Commercially available biotinylated anti-Cy3 antibody was used to label the chromophore of DiI-C18 on the young thrombocytes and streptavidin coated magnetic beads were added subsequently, to separate young thrombocytes. The remaining blood cells were probed with custom-made biotinylated anti-GPIIb antibody and streptavidin magnetic beads to separate them from other cells. Further, thrombocytes are equivalents of mammalian platelets. Platelets play a crucial role in thrombus formation. The G-protein coupled receptors (GPCRs) present on the platelet surface are involved during platelet activation and aggregation processes. So, thrombocytes were studied for the presence of GPCRs. The GPCR mRNA transcripts expressed in the young and mature thrombocytes were subjected to densitometry analysis and pixel intensities of the bands were compared using one way ANOVA. This analysis did not show significant differences between the young and mature GPCR mRNA transcripts but identified a novel GPCR, GPR18 that was not reported in platelets earlier. To study the function of this GPCR, it was knocked down using GPR18 specific antisense morpholino and vivo morpholino. The immunofluorescence experiment indicated the presence of GPR18 on thrombocytes. The results of the assays, such as, time to occlusion (TTO) and time to aggregation (TTA) in response to N-arachidonyl glycine (NAG) as an agonist, showed prolongation of time in GPR18 larval and adult morphants respectively, suggesting that GPR18 plays a role in thrombus formation in zebrafish. In conclusion, our results indicate that GPR18 may be present in zebrafish thrombocytes, it may be involved in thrombus formation and that NAG may be an agonist at GPR18 on thrombocytes.
Date: May 2013
Creator: Potbhare, Vrinda Nikhil
Partner: UNT Libraries

Molecular and Functional Characterization of Medicago Truncatula Npf17 Gene

Description: Legumes are unique among plants for their ability to fix atmospheric nitrogen with the help of soil bacteria rhizobia. Medicago truncatula is used as a model legume to study different aspects of symbiotic nitrogen fixation. M. truncatula, in association with its symbiotic partner Sinorhizobium meliloti, fix atmospheric nitrogen into ammonia, which the plant uses for amino acid biosynthesis and the bacteria get reduced photosynthate in return. M. truncatula NPF1.7 previously called MtNIP/LATD is required for symbiotic nitrogen fixing root nodule development and for normal root architecture. Mutations in MtNPF1.7 have defects in these processes. MtNPF1.7 encodes a member of the NPF family of transporters. Experimental results showing that MtNPF1.7 functioning as a high-affinity nitrate transporter are its expression restoring chlorate susceptibility to the Arabidopsis chl1-5 mutant and high nitrate transport in Xenopus laevis oocyte system. However, the weakest Mtnip-3 mutant allele also displays high-affinity nitrate transport in X. laevis oocytes and chlorate susceptibility to the Atchl1-5 mutant, suggesting that MtNPF1.7 might have another biochemical function. Experimental evidence shows that MtNPF1.7 also functions in hormone signaling. Constitutive expression of MtNPF1.7 in several species including M. truncatula results in plants with a robust growth phenotype. Using a synthetic auxin reporter, the presence of higher auxin in both the Mtnip-1 mutant and in M. truncatula plants constitutively expressing MtNPF1.7 was observed. Previous experiments showed MtNPF1.7 expression is hormone regulated and the MtNPF1.7 promoter is active in root and nodule meristems and in the vasculature. Two potential binding sites for an auxin response factors (ARFs) were found in the MtNPF1.7 promoter. Chromatin immunoprecipitation-qRT-PCR confirmed MtARF1 binding these sites. Mutating the MtARF1 binding sites increases MtNPF1.7 expression, suggesting a mechanism for auxin repression of MtNPF1.7. Consistent with these results, constitutive expression of an ARF in wild-type plants partially phenocopies Mtnip-1 mutants’ phenotypes.
Date: December 2013
Creator: Salehin, Mohammad
Partner: UNT Libraries

Origin and Role of Factor Viia

Description: Factor VII, the initiator of the extrinsic coagulation cascade, circulates in human plasma mainly in its zymogen form, Factor VII and in small amounts in its activated form, Factor VIIa. However, the mechanism of initial generation of Factor VIIa is not known despite intensive research using currently available model systems. Earlier findings suggested serine proteases Factor VII activating protease, and hepsin play a role in activating Factor VII, however, it has remained controversial. In this work I estimated the levels of Factor VIIa and Factor VII for the first time in adult zebrafish plasma and also reevaluated the role of the above two serine proteases in activating Factor VII in vivo using zebrafish as a model system. Knockdown of factor VII activating protease did not reduce Factor VIIa levels while hepsin knockdown reduced Factor VIIa levels. After identifying role of hepsin in Factor VII activation in zebrafish, I wanted to identify novel serine proteases playing a role in Factor VII activation. However, a large scale knockdown of all serine proteases in zebrafish genome using available knockdown techniques is prohibitively expensive. Hence, I developed an inexpensive gene knockdown method which was validated with IIb gene knockdown, and knockdown all serine proteases in zebrafish genome. On performing the genetic screen I identified 2 novel genes, hepatocytes growth factor like and prostasin involved in Factor VII activation.
Date: December 2013
Creator: Khandekar, Gauri
Partner: UNT Libraries

Functional Characterization of Mtnip/latd’s Biochemical and Biological Function

Description: Symbiotic nitrogen fixation occurs in plants harboring nitrogen-fixing bacteria within the plant tissue. The most widely studied association is between the legumes and rhizobia. In this relationship the plant (legumes) provides the bacteria (rhizobia) with reduced carbon derived from photosynthesis in exchange for reduced atmospheric nitrogen. This allows the plant to survive in soil, which is low in available of nitrogen. Rhizobia infect and enter plant root and reside in organs known as nodules. In the nodules the bacteria fix atmospheric nitrogen. The association between the legume, Medicago truncatula and the bacteria Sinorhizobium meliloti, has been studied in detail. Medicago mutants that have defects in nodulation help us understand the process of nitrogen fixation better. One such mutant is the Mtnip-1. Mtnip-1 plants respond to S. meliloti by producing abnormal nodules in which numerous aberrant infection threads are produced, with very rare rhizobial release into host plant cells. The mutant plant Mtnip-1 has an abnormal defense-like response in root nodules as well as defects in lateral root development. Three alleles of the Mtnip/latd mutants, Mtnip-1, Mtlatd and Mtnip-3 show different degrees of severity in their phenotype. Phylogenetic analysis showed that MtNIP/LATD encodes a protein belonging to the NRT1(PTR) family of nitrate, peptide, dicarboxylate and phytohprmone transporters. Experiments with Mtnip/latd mutants demonstrats a defective nitrate response associated with low (250 μM) external nitrate concentration rather than high (5 mM) nitrate concentration. This suggests that the mutants have defective nitrate transport. To test if MtNIP/LATD was a nitrate transporter, Xenopus laevis oocytes and Arabidopsis thaliana mutant plants Atchl1-5, defective in a major nitrate transporter AtNRT1.1(CHL1), were used as surrogate expression systems. Heterologous expression of MtNIP/LATD in X. laevis oocytes and Atchl1-5 mutant plants conferred on them the ability to take up nitrate from external media with high affinity, thus demonstrating that MtNIP/LATD ...
Date: December 2013
Creator: Bagchi, Rammyani
Partner: UNT Libraries