Location: Agricultural Genetic Resources Preservation Research
Title: Tracking the permeation of PVS2 components in living Mentha × piperita shoot tips using optical microscopiesAuthor
KRECKEL, H - Colorado State University | |
OCHOA CASTILLO, A - Colorado State University | |
STICH, D - University Of Colorado | |
Bonnart, Remi | |
Volk, Gayle | |
LEVINGER, N - Colorado State University |
Submitted to: Society for Cryobiology Meeting
Publication Type: Abstract Only Publication Acceptance Date: 7/25/2023 Publication Date: N/A Citation: N/A Interpretive Summary: N/A Technical Abstract: Cryopreservation of vegetatively propagated crop collections has emerged as a solution to the global plant conservation crisis. The successful cryopreservation of these collections relies upon the exposure of 1-2 mm shoot tips to highly concentrated mixtures of cryoprotective agents (CPAs) to reduce water content and increase the viscosity of cells prior to liquid nitrogen exposure. Despite the successful use of CPA mixtures such as PVS2 (15% w/v dimethyl sulfoxide, 15% w/v ethylene glycol, 30% w/v glycerol and 0.4 M sucrose), they are not universally cryoprotective and very little is understood about how these CPAs protect cells and tissues from freezing damage. Coherent anti-Stokes Raman scattering (CARS) microscopy is a non-invasive vibrational imaging technique that detects a single chemical bond of interest, allowing for direct visualization of CPAs within living plant cells. Previously, we have presented results pairing CARS microscopy with both brightfield and two-photon endogenous fluorescence microscopies to explore the extent of Me2SO4 permeation within peppermint shoot tips. This presentation extends our exploration to ethylene glycol and glycerol components of PVS2, providing a comprehensive picture of how permeating components of PVS2 localize in peppermint shoot tips. Further, we will present the latest results of immunofluorescent tissue staining, providing an even more comprehensive view of tissue development and cellular components. |