CHARACTERIZATION OF TOBACCO MOSAIC VIRUS DIRECTED REPROGRAMMING OF PHLOEM GENE EXPRESSION TO PROMOTE VIRAL PHLOEM LOADING AND SYSTMEIC MOVEMENT

Vascular phloem loading has long been recognized as an essential step in the establishment of a systemic virus infection. Yet little is known about this process and the mechanisms that control it. In this study, an interaction between the replication protein of Tobacco mosaic virus (TMV) and phloem specific auxin/indole acetic acid (Aux/IAA) transcriptional regulators was found to modulate virus phloem loading. Promoter expression studies show TMV 126/183 kDa interacting Aux/IAAs predominantly express and accumulate within the nuclei of phloem companion cells (CC). Furthermore, CC Aux/IAA nuclear localization is disrupted upon infection with an interacting virus but not during infection with a non-interacting virus. In situ analysis of virus spread shows the inability of TMV variants to disrupt Aux/IAA CC nuclear localization correlates with a reduced ability to load into the vascular tissue. Subsequent systemic movement assays also demonstrate that a virus capable of disrupting Aux/IAA localization is significantly more competitive at systemic movement than a non-interacting virus. Similarly, CC expression and over-accumulation of a degradation-resistant-interacting Aux/IAA protein was found to selectively inhibit TMV accumulation and phloem loading. Transcriptional expression studies demonstrate a role for interacting Aux/IAA proteins in the regulation of salicylic acid and jasmonic acid dependent host defense responses as well as virus specific movement factors including pectin methylesterase that are involved in regulating plasmodesmata size exclusion limits and promoting virus cell-to-cell movement. Further characterization of the phloem environment was done using two phloem specific promoters (pSUC2 and pSULTR2;2) to generate epitope-tagged polysomal-RNA complexes. Immuno-purification using the epitope tag allowed us to obtain mRNAs bound to polysomes (the translatome) specifically in phloem tissue. We found the phloem translatome is uniquely altered during TMV infection with 90% and 88% of genes down regulated in the pSUC2 and pSULTR2;2 phloem translatomes, compared to 31% of genes down regulated in the whole plant p35S translatome. Transcripts down regulated in phloem include genes involved in callose deposition at plasmodesmata, host defense responses, and RNA silencing. Combined, these findings indicate TMV reprograms gene expression within the vascular phloem as a means to enhance phloem loading and systemic spread.

Essays in Greenhouse/Nursery Economics

This dissertation focuses on the greenhouse and nursery industry in the United States. Two major issues are explored: irrigation and plant disease. The first two essays examine wireless soil-moisture sensor networks, an emerging technology that measures soil moisture and optimizes irrigation levels in real time. The first essay describes a study in which a nationwide survey of commercial growers was administered to generate estimates of grower demand and willingness to pay for sensor networks. We find that adoption rates for a base system and demand for expansion components are decreasing in price, as expected. The price elasticity of the probability of adoption suggests that sensor networks are likely to diffuse at a rate somewhat greater than that of drip irrigation. In the second essay, yields, time-to-harvest, and plant quality were analyzed to measure sensor network profitability. Sensor-based irrigation was found to increase revenue by 62% and profit by 65% per year. The third essay investigates greenhouse nursery growers’ response to a quarantine imposed on the west coast of the United States from 2002 to present for the plant pathogen that causes Sudden Oak Death. I investigate whether growers choose to 1) improve their sanitation practices, which reduces the underlying risk of disease without increasing the difficulty of detecting the pathogen, 2) increase fungicide use, which also prevents disease but makes existing infections much harder to detect, or 3) change their crop composition towards more resistant species. First, a theoretical model is derived to formalize hypotheses on grower responses to the quarantine, and then these predictions are empirically tested using several public data sources. I do not find evidence that growers improve their sanitation practices in response to the quarantine. I do, however, find evidence that growers heavily increase their fungicide use in response to a quarantine policy that requires visual (as opposed to laboratory) inspection for the disease before every crop shipment, suggesting that the quarantine may have the adverse effect of making the pathogen harder to identify. I also do find evidence that growers shift away from susceptible crops and towards resistant crops.