No description available.

No description available.

No description available.

No description available.

Rac/Rop GTPases are molecular switches in plants that control the growth of polarized cells such as pollen tubes and root hairs, differentiation, development, actin dynamics, production of reactive oxygen species (ROS) and disease resistance. These small GTPases are activated by guanine nucleotide exchange factors (GEFs) that replace GDP for GTP and are referred to as RopGEFs in plants. To identify upstream components of the RopGEF regulated signaling pathways, GEF1 from Arabidopsis thaliana was used as a bait to screen a seedling cDNA library in a yeast two-hybrid system. This yielded members of a small family of the Catharanthus roseus Receptor-like kinase (CrRLK), referred to as the Feronia-like Receptor-like kinase (FlRLK) family as the potential GEF interactors. A synergid cell-expressed member of this family, FERONIA/SIRENE (RLK-10), regulates pollen tube reception by the female gametophyte and along with the other two plasma-membrane bound receptors of this family, THESEUS (RLK-3) and HERCULES (RLK-4) promote cell elongation in Arabidopsis. I have chosen two of these FlRLKs for my studies: RLK-5, the most abundant pollen expressed member of the family and RLK10. My studies with RLK-5 suggest that it plays a significant role in pollination or fertilization since homozygous rlk-5 was never recovered from selfed heterozygous rlk-5 mutant. In addition to this, the reciprocal crosses with RLK-5/rlk-5 and wild type resulted in severe male transmission defect indicating that the rlk-5 mutation induces male sterility. This observation was found consistent with the pollen-specific expression pattern of RLK-5 suggested by microarray data and confirmed by histochemical GUS staining analysis of the RLK5p-GUS transgenic Arabidopsis. The RLK-5/rlk-5 pollen displayed no pollen-viability defects and the pollen tube growth in-vivo appears normal. Despite the unaffected pollen viability and apparent normal in-vivo pollen tube growth, the RLK-5/rlk-5 plants formed low seed set suggesting compromised fertilization. Additional analysis will be required to determine the basis of male deficiency and reduced seed sets in RLK5/rlk5 mutants. Loss of function mutations of RLK-10 (FERONIA/SIRENE) gene show failure in pollen tube growth arrest upon penetration of the female gametophyte, supernumerary pollen tube penetration of the ovule and reduced female fertility. Our studies with RLK-10 suggest that it is important for mediating a proper oxido-reductive condition within the ovule necessary for pollen tube rupture and fertilization of the female gametophyte. The multiple pollen tube entry phenotype of the Arabidopsis knockout mutant, rlk-10 correlated with decreased ROS level and de-esterified pectin in the filiform apparatus lining the synergid cell of its female gametophyte. These results establish an important signaling link between RLK-10 and ROS and pectin in a GEF-Rac/Rop regulated pathway in pollen tube-ovule interaction.

Dodder (Cuscuta gronovii Willd.) is a weed of serious concern to cranberry (Vaccinium macrocarpon Ait.) growers. It develops vigorously and has a long-lived seed bank. Cranberries are a perennial crop and therefore strategies available to growers of annual crops are not practical. Herbicides, the primary management tool for dodder, although effective, have a narrow window of application and extended seedling emergence after applications can result in escapes. This project examined the effect of water temperature on dodder seed germination and the use of short-term floods (less than 72 hr) for dodder management. Experiments investigated the effect of water temperature on dodder seed germination. Studies, ran twice, submerged dodder seed in water for 0 to 48 hr at 10, 15, and 20 C in one experiment (simulating spring water temperatures) and 0 to 48 hr at 15, 20, and 25 C in a separate experiment (simulating summer water temperatures). In Run 1, the effect of temperature on percent seed germination varied by flood duration; and by temperature alone in Run 2. Percent seed germination however, always fell within normal ranges (35-59%), indicating that flooding may not impact seed germination. Two 1-yr field studies were conducted to evaluate the use of short-term floods (24 to 48 hr) for managing dodder in cranberries. Two scenarios were simulated: cranberry beds with no emergent weed populations (cranberries alone) and cranberries with emergent weed populations (cranberries with additional host). There were three flood durations (0, 24, and 48 hr) and four flood initiations (1 to 4 wk after first seedling emergence). In 2006, mean percent germination from seeds incubated in Petri dishes was lower for seeds submerged 3 and 4 wk after first emergence (AFE) for the 48-hr flood durations. In 2007, mean percent germination for seeds submerged for 24 and 48-hr decreased for floods initiated at 4 wk AFE. Flooding 4 wk AFE resulted in lowest mean attachment ratings in both years and lowest mean dodder biomass on cranberry in the 2007 cranberry and tomato study, suggesting later flood initiation may provide better dodder management.

The regulation of pH is a critical homeostatic function of plant cells. In addition to acting as the primary cationic species responsible for energizing the plasma membrane, protons likely act as an important regulator and messenger. Despite this importance, few studies have thoroughly described cytosolic pH patterns as the plant cell progresses through the cell cycle. To investigate pH in plant cells, I chose Nicotiana tabacum (tobacco) Bright Yellow-2 (BY-2) cells as a model system. My research has two aims. First, I will measure and report the interphase cytosolic pH of BY-2 cells. Next, I will assay the cytosolic pH as BY-2 cells progress through mitosis and cytokinesis. I hypothesize that pH patterns are be temporally or spatially associated with structures such as the mitotic spindle or the phragmoplast. To investigate cytosolic pH in BY-2 cells, I will develop a cytosolic pH reporter based on a pH sensitive ratiometric fluorescent dye. This dye will be able to resolve both temporal and spatial changes in pH throughout the cytosol while imposing a minimal amount of stress on BY-2 cells. I found that pH-GFP, a variant of eGFP, had qualities of a robust pH reporter. To introduce the dye, explored biolistic bombardment, Agrobacterium mediated transient transformation, and polyethylene glycol mediated transformation as methods for introducing the pH-GFP gene into BY-2 cells. I observed very few transformation events using these methods and my observations did not support these approaches as suitable for introducing pH-GFP into BY-2 cells.

The geographic mosaic theory of coevolution predicts that spatial differences in species interactions result in a patchwork of evolutionary hot and cold spots across a landscape. We used horsenettle (Solanum carolinense L.), a perennial weed with a diverse insect community found in old fields and meadows, to examine local adaptation and resource-mediated selection. The goals of this study were to (1) determine the potential for a selection mosaic by identifying local adaptation through trait-interaction matching with herbivores, pollinations and plant competitors, and (2) determine the potential for indirect selection through resource allocation tradeoffs. The potential for local adaptation was determined by measuring interactions in four populations and relating those findings to plant traits measured on offspring grown from those populations in a ‘common garden.’ Allocation tradeoffs between growth, herbivore resistance, and floral traits were also assessed in the common garden. We found high herbivore damage in the field associated with decreased root:shoot ratios in greenhouse-grown plants, which may indicate an herbivore-mediated effect on life-history through selection for a more annual strategy. By examining allocation tradeoffs we found evidence of two distinct reproductive strategies in this perennial plant. Negative correlations between reproductive traits and both growth and defense suggest that individuals either favor current growth and reproduction over defense, or invest in current survival and defense while delaying reproduction. Overall, this study sheds light on how selection changes over space and time, which are of many of the fascinating traits we find in plants and animals today.

The progressive increase in the global population and the rapidly changing climate have put unprecedented pressure on crop production. Cowpea is one of the world’s most important leguminous crops, contributing to food security and environmental sustainability. However, cowpea productivity is limited due to waterlogging stress. The main objective of this study was to explore physiological and biochemical mechanisms to understand how cowpea genotypes respond to waterlogging stress. Four studies were conducted in controlled and field conditions to achieve these objectives. Study 1 characterized the waterlogging tolerance of 30 cowpea genotypes in a controlled environment using 24 morphophysiological parameters with waterlogging tolerance coefficients and multivariate analysis methods. 10% of the genotypes exhibited high tolerance to waterlogging stress, and the genotypes UCR 369 and EpicSelect.4 were identified as the most and least waterlogging tolerant, respectively. Study 2 evaluated the key parameters influencing carbon fixation of UCR 369 and EpicSelect.4 at the reproductive stage. The less tolerant EpicSelect.4 experienced high downregulation of stomatal and non-stomatal limiting factors during waterlogging and recovery, resulting in decreased carbon assimilation rates. UCR 369 rapidly developed adventitious roots, maintained biomass, and restored pigments and metabolites to sustain photosynthesis. A two-year field experiment was conducted in study 3 to quantify the effects of waterlogging on the yields, physiology, and biochemistry of cowpeas at different growth stages. The most apparent impact of waterlogging stress occurred at the reproductive stage, followed by the vegetative and maturity growth stages. Studies suggest that diverse cowpea genotypes have distinct physiological and biochemical mechanisms in response to waterlogging stress. In addition, the tolerant genotypes and traits identified herein can be used in genetic engineering and cowpea breeding programs that integrate increased yield with waterlogging stress tolerance.

Suboptimal soil moisture during the growing season often limits growth and yield potential of corn. This study aimed to assess the impact of varying soil moisture regimes on corn growth at different growth phases involving vegetative, flowering, and grain-filling stages. Exposure to moisture stress (80% of the control) during the vegetative stage resulted in a 65% reduction in stomatal conductance and increased the canopy temperature by 5 oC, which led to a substantial decrease in total dry matter (56%). Moisture stress-induced reductions in silk length (19%) and fresh weight (34%), negatively affected kernel number (53%), and weight (54%). Unlike the flowering stage, extreme stress during grain-filling had a greater impact on kernel weight (19%) than the number (7%). During flowering, stress reduced kernel starch content with an increase in protein content. Our findings infer that improving the resilience of the corn flowering stage to soil moisture stress may help reduce the yield gap between irrigated and rainfed.