Low bioavailability of phosphate (P) due to low concentration and high immobility in soils is a key limiting factor in crop production. Application of excess amounts of P fertilizer is costly and by no means sustainable, as world-wide P resources are finite and running out. To facilitate the breeding of crops adapted to low-input soils, it is essential to understand the consequences of P deficiency. The second messenger calcium (Ca2+) is known to signal in plant development and stress perception, and most recently its direct role in signalling nutrient availability and deficiency has been partially elucidated. The use of Ca2+ as a signal has to be tightly controlled, as Ca2+ easily complexes with P groups and therefore is highly toxic to cellular P metabolism. It is unknown whether Ca2+ signals P availability or whether signalling is altered under P starvation conditions. The aim of this PhD project was to characterise the use of Ca2+ ions, particularly cytosolic free Ca2+ ([Ca2+]cyt), in stress signalling by P-starved roots of the model plant Arabidopsis thaliana. The hypothesis was that under P starvation and a resulting decreased cellular P pool, the use of [Ca2+]cyt may have to be restricted to avoid cytotoxic complexation of Ca2+ with limited P groups. Employing a range of genetically encoded Ca2+ reporters in Arabidopsis, P starvation but not nitrogen starvation was found to strongly dampen the root [Ca2+]cyt increases evoked by mechanical, salt, osmotic, and oxidative stress as well as by extracellular nucleotides. The strongly altered root [Ca2+]cyt response to extracellular nucleotides was shown to manifest itself during seedling development under chronic P deprivation, but could be reversed by P resupply. Fluorescent imaging elucidated that P-starved roots showed a normal [Ca2+]cyt response to extracellular nucleotides at the apex, but a strongly dampened [Ca2+]cyt response in distal parts of the root tip, correlating with high reactive oxygen species (ROS) levels induced by P starvation. Excluding iron, as well as P, rescued the altered [Ca2+]cyt response, and restored ROS levels to those seen under nutrient-replete conditions. P availability was not signalled through [Ca2+]cyt. In another part of this PhD project, a library of 77 putative Ca2+ channel mutants was compiled and screened for aberrant root hair growth under P starvation conditions. No mutant line showed aberrant root hair growth. These results indicate that P starvation strongly affects stress-induced [Ca2+]cyt modulations. The data generated in this thesis further understanding of how plants can integrate nutritional and environmental cues, adding another layer of complexity to the use of Ca2+ as a signal transducer.

<p>Plant root phenotyping technologies play an important role in breeding, plant protection, and other plant science research projects. The root phenotyping customers urgently need technologies that are low-cost, in situ, non-destructive to the roots, and suitable for the natural soil environment. Many recently developed root phenotyping methods such as minirhizotron, CT, and MRI scanners have their unique advantages in observing plant roots, but they also have disadvantages and cannot meet all the critical requirements simultaneously. The study in this paper focuses on the development of a new plant root phenotyping robot that is minimally invasive to plants and working in situ inside natural soil, called “MISIRoot”. The MISIRoot system (patent pending) mainly consists of an industrial-level robotic arm, a mini-size camera with lighting set, a plant pot holding platform, and the image processing software for root recognition and feature extraction. MISIRoot can take high-resolution color images of the roots in soil with minimal disturbance to the root and reconstruct the plant roots’ three-dimensional (3D) structure at an accuracy of 0.1 mm. In a test assay, well-watered and drought-stressed groups of corn plants were measured by MISIRoot at V3, V4, and V5 stages. The system successfully acquired the RGB color images of the roots and extracted the 3D points cloud data which showed the locations of the detected roots in the soil. The plants measured by MISIRoot and plants not measured (controls) were carefully compared with Purdue’s Lilly 13-4 Hyperspectral Imaging Facility (reference). No significant differences were found between the two groups of plants at different growth stages. Therefore, it was concluded that MISIRoot measurements had no significant disturbance to the corn plant’s growth.</p>

Manure contains nutrients for crop growth; however, overapplication, with time, can result in excess nutrients in soil, which can subsequently be lost in surface runoff. <p>The general purpose of this research is to study the effect of liquid hog manure, applied as an agricultural fertilizer, on water chemistry of surface runoff from rainfall simulation tests. Specifically the research focuses on runoff water chemistry comparisons between lands receiving hog manure at different rates, via different injection methods, and upon different slope positions. <p>To examine these objectives, soil nutrient supply rates (P, NH4-N, and NO3-N) of the 0 5 cm depth of soil adjacent to rainfall simulation positions, and runoff water chemistry (TP, OP, NH4-N, NO3-N, DOC, Cl- and coliforms) during rainfall simulation tests were collected before and after manure addition. <p> Generally, manure application did increase soil NH4-N and NO3-N supply rates, and runoff NH4-N concentration. Soil P supply rate and runoff TP concentration were not affected by the manure addition; however, runoff OP concentration at one site (Perdue) increased significantly due to manure addition. The manure treatments applied in this study did not cause any significant increases in fecal or total coliform in runoff from rainfall simulation tests conducted 7 8 months after manure application. None of the water quality parameters exceeded the Guidelines for Canadian Drinking Water Quality. <p> Manure injection method (regular versus low soil surface disturbance) had consistent effects on runoff chemistry, but application rate did not. The regular disturbance method had significantly higher concentrations of water quality parameters than the low disturbance method. <p> The position of the test on the slope did not result in any consistent trends in runoff chemistry, whether before or after manure addition. Foot slope positions had higher soil NH4-N supply rates than upper slope positions, both before and after manure addition. Soil NH4-N, NO3-N, and P supply rates between landscape positions were not likely influenced by manure addition. <p> Regression tests between soil nutrient supply rates and runoff chemistry indicate that soil NH4-N supply rates are a good index to predict runoff NH4-N concentration, but soil P did not predict runoff P.

Manure contains nutrients for crop growth; however, overapplication, with time, can result in excess nutrients in soil, which can subsequently be lost in surface runoff. <p>The general purpose of this research is to study the effect of liquid hog manure, applied as an agricultural fertilizer, on water chemistry of surface runoff from rainfall simulation tests. Specifically the research focuses on runoff water chemistry comparisons between lands receiving hog manure at different rates, via different injection methods, and upon different slope positions. <p>To examine these objectives, soil nutrient supply rates (P, NH4-N, and NO3-N) of the 0 5 cm depth of soil adjacent to rainfall simulation positions, and runoff water chemistry (TP, OP, NH4-N, NO3-N, DOC, Cl- and coliforms) during rainfall simulation tests were collected before and after manure addition. <p> Generally, manure application did increase soil NH4-N and NO3-N supply rates, and runoff NH4-N concentration. Soil P supply rate and runoff TP concentration were not affected by the manure addition; however, runoff OP concentration at one site (Perdue) increased significantly due to manure addition. The manure treatments applied in this study did not cause any significant increases in fecal or total coliform in runoff from rainfall simulation tests conducted 7 8 months after manure application. None of the water quality parameters exceeded the Guidelines for Canadian Drinking Water Quality. <p> Manure injection method (regular versus low soil surface disturbance) had consistent effects on runoff chemistry, but application rate did not. The regular disturbance method had significantly higher concentrations of water quality parameters than the low disturbance method. <p> The position of the test on the slope did not result in any consistent trends in runoff chemistry, whether before or after manure addition. Foot slope positions had higher soil NH4-N supply rates than upper slope positions, both before and after manure addition. Soil NH4-N, NO3-N, and P supply rates between landscape positions were not likely influenced by manure addition. <p> Regression tests between soil nutrient supply rates and runoff chemistry indicate that soil NH4-N supply rates are a good index to predict runoff NH4-N concentration, but soil P did not predict runoff P.