J Stock1, A Semula2, M Nagel2, H Mock2, E R Joachim Keller2. 1. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany. stock@ipk-gatersleben.de. 2. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany.
Abstract
BACKGROUND: The use of the model plant Arabidopsis could be a valuable tool to elucidate the basic mechanisms involved in plant cryopreservation. OBJECTIVE: A simple and powerful protocol, independent of Arabidopsis genotypes, was established using a PVS2 protocol. MATERIALS AND METHODS: Two PVS2 (a and b), one PVS3 droplet-vitrification and one DMSO droplet-freezing protocol were tested with alternating temperatures during the growing phase of donor plants. RESULTS: PVS2 protocols, including cold acclimation of donor plants, resulted in highest recovery. The PVS2a protocol was successfully applied to a collection of different Arabidopsis genotypes with an average recovery of 94%. In addition, Differential Scanning Calorimetry confirmed the occurrence of glass transitions in the PVS3 and PVS2 protocols. CONCLUSION: The PVS2a protocol is suitable to screen the large collection of Arabidopsis mutants and transgenic lines with the aim to identify cellular functions associated with cryopreservation tolerance.
BACKGROUND: The use of the model plant Arabidopsis could be a valuable tool to elucidate the basic mechanisms involved in plant cryopreservation. OBJECTIVE: A simple and powerful protocol, independent of Arabidopsis genotypes, was established using a PVS2 protocol. MATERIALS AND METHODS: Two PVS2 (a and b), one PVS3 droplet-vitrification and one DMSO droplet-freezing protocol were tested with alternating temperatures during the growing phase of donor plants. RESULTS: PVS2 protocols, including cold acclimation of donor plants, resulted in highest recovery. The PVS2a protocol was successfully applied to a collection of different Arabidopsis genotypes with an average recovery of 94%. In addition, Differential Scanning Calorimetry confirmed the occurrence of glass transitions in the PVS3 and PVS2 protocols. CONCLUSION: The PVS2a protocol is suitable to screen the large collection of Arabidopsis mutants and transgenic lines with the aim to identify cellular functions associated with cryopreservation tolerance.
Authors: Johanna Stock; Andrea Bräutigam; Michael Melzer; Gerd Patrick Bienert; Boyke Bunk; Manuela Nagel; Jörg Overmann; E R Joachim Keller; Hans-Peter Mock Journal: J Exp Bot Date: 2020-08-06 Impact factor: 6.992
Authors: Felizitas Bajerski; Johanna Stock; Benjamin Hanf; Tatyana Darienko; Elke Heine-Dobbernack; Maike Lorenz; Lisa Naujox; E R J Keller; H M Schumacher; Thomas Friedl; Sonja Eberth; Hans-Peter Mock; Olaf Kniemeyer; Jörg Overmann Journal: Front Physiol Date: 2018-07-17 Impact factor: 4.566