Investigating the function of Arabidopsis KOD ; and, Developing a microcontroller based system for continuous and high throughput quantification of plant cell death by ion leakage

Endeavours to advance understanding of KISS OF DEATH (KOD) gene function by established methods were taken forward. Particularly, by attempting to utilise the previously reported ability of ectopic KOD expression to induce Programmed Cell Death (PCD). To this end, KOD was expressed in Arabidopsis and tobacco. However the induction of a PCD phenotype by KOD could not be replicated in this work. As a consequence, efforts towards the aim of understanding how KOD induced PCD were inhibited. Some understanding of the biology of KOD function was gained by the investigation of KOD transcription. It was found that low level KOD expression could be induced by treatment with 500mM sodium chloride, in addition to the already characterised induction of KOD by H2O2, heat shock and Pseudomonas syringae AvrRPM1. This led to the hypothesis proposed here that KOD induction in response to stress treatments may be incidental to, rather than a contributory factor to, cell death. This is based on analysis of response to H2O2 and Pseudomonas syringae AvrRPM1 in kod mutants. Furthermore, a pKOD:GUS reported was developed and characterised. Taken together, evidence obtained here eroded support for perceiving KOD as a potent core component contributing to multiple PCD pathways. The design, construction, calibration and validation of a successful new conductivity meter system is presented. This was designed to update and improve the classic electrolyte leakage assay for cell death. The device is able to record measurements from a large number of individual samples simultaneously and continuously, without requiring further operator intervention. The device conforms to the principles of the open source hardware movement and the standards of the open-source Arduino micro-controller platform.