title: Freezing behaviours in wintering Cornus florida flower bud tissues revisited using MRI.
abstract: How plant tissues control their water behaviours (phase and movement) under subfreezing temperatures through adaptative strategies (freezing behaviours) is important for their survival. However, the fine details of freezing behaviours in complex organs and their regulation mechanisms are poorly understood, and non-invasive visualization/analysis is required. The localization/density of unfrozen water in wintering Cornus florida flower buds at subfreezing temperatures was visualized with high-resolution magnetic resonance imaging (MRI). This allowed tissue-specific freezing behaviours to be determined. MRI images revealed that individual anthers and ovules remained stably supercooled to -14 to -21 °C or lower. The signal from other floral tissues decreased during cooling to -7 °C, which likely indicates their extracellular freezing. Microscopic observation and differential thermal analyses revealed that the abrupt breakdown of supercooled individual ovules and anthers resulted in their all-or-nothing type of injuries. The distribution of ice nucleation activity in flower buds determined using a test tube-based assay corroborated which tissues primarily froze. MRI is a powerful tool for non-invasively visualizing unfrozen tissues. Freezing events and/or dehydration events can be located by digital comparison of MRI images acquired at different temperatures. Only anthers and ovules preferentially remaining unfrozen are a novel freezing behaviour in flower buds. Physicochemical and biological mechanisms/implications are discussed.
authors: Division of Plant Sciences, National Institute of Agrobiological Sciences, Kan'nondai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan.|Imaging Frontier Center, Tokyo University of Science, Yamazaki 2641, Noda, Chiba, 278-8510, Japan.|Department of Forest Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors: Masaya
authors: M
authors: Ishikawa
authors: Water Research Institute, Sengen 2-1-6, Tsukuba, Ibaraki, 305-0047, Japan.|Institute for Innovation, Ajinomoto Co., Inc., Suzuki 1-1, Kawasaki, Kawasaki, 210-8681, Japan.
authors: Hiroyuki
authors: H
authors: Ide
authors: Division of Plant Sciences, National Institute of Agrobiological Sciences, Kan'nondai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan.|International Patent Organism Depository, National Institute of Technology and Evaluation, Kisarazu, Chiba, 292-0818, Japan.
authors: Hideyuki
authors: H
authors: Yamazaki
authors: Division of Plant Sciences, National Institute of Agrobiological Sciences, Kan'nondai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan.|Department of Applied Biological Science, Tokyo University of Science, Yamazaki 2641, Noda, Chiba, 278-8510, Japan.
authors: Hiroki
authors: H
authors: Murakawa
authors: Department of Applied Biological Science, Tokyo University of Science, Yamazaki 2641, Noda, Chiba, 278-8510, Japan.|Imaging Frontier Center, Tokyo University of Science, Yamazaki 2641, Noda, Chiba, 278-8510, Japan.
authors: Kazuyuki
authors: K
authors: Kuchitsu
authors: Water Research Institute, Sengen 2-1-6, Tsukuba, Ibaraki, 305-0047, Japan.|Nanoscale Organisation and Dynamics Group, Western Sydney University, Penrith, New South Wales, 2751, Australia.
authors: William S
authors: WS
authors: Price
authors: Water Research Institute, Sengen 2-1-6, Tsukuba, Ibaraki, 305-0047, Japan.
authors: Yoji
authors: Y
authors: Arata
mesh_terms: D029768:Cornus / Q000033:anatomy & histology / Q000502:physiology*; D035264:Flowers / Q000033:anatomy & histology / Q000502:physiology* / Q000648:ultrastructure; D005615:Freezing / Q000009:adverse effects; D008279:Magnetic Resonance Imaging; D008853:Microscopy
keywords: anther; cold hardiness; deep supercooling; freezing injury; nuclear magnetic resonance (NMR) micro-imaging; ovule