A MECHANISTIC APPROACH IMPROVING CRYOPRESERVATION TECHNOLOGIES TO PRESERVE AND PROTECT AMERICAN AGRICULTURE

The research proposed here will revolutionize plant cryopreservation through the application of state-of-the-art coherent Raman vibrational microscopyand foundational spectroscopic studies that will provide quantitative data about how and why cryopreservation solutions function and enable improved plant cryopreservation by tailoring solutions to plant specific applications. Because these formulations work "well enough" in many applications, only a few studies have attempted to evaluate how and why these formulations succeed or fail.We willalso apply methods we develop to explore the mechanism of plant cryotherapy, that is,cryogenic conditions applied to plant shoot tipsto eradicate pathogens from clonal propagules. Like cryopreservation, the biochemical and biophysical mechanisms of plant cryotherapy remain poorly understood. We will use methods developed for cryopreservation studies to develop quantitative knowledge about plant cryotherapy.Specific objectives include:To develop quantitative tools that enable direct and universal measurement of permeating and non-permeating cryoprotectants in plant cells and shoot tips.To discover fundamental structural and temporal characteristics associated with permeating and non-permeating cryoprotecting agents that lead to effective plant cryopreservation.To localize bacteria within shoot tips and directly measure the effect of liquid nitrogen exposure on their eradication in the cryotherapy process.