Screening of Ice Nucleation Substances in Cold Hardy Dwarf Bamboos

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  • KUBO Hikaru
    Department of Forest Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo
  • ISHIKAWA Masaya
    Department of Forest Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo
  • MATSUSHITA Norihisa
    Department of Forest Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo
  • FUKUDA Kenji
    Department of Forest Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo

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Other Title
  • ササ類に含まれる氷核活性物質の探索
  • ササルイ ニ フクマレル ヒョウカク カッセイ ブッシツ ノ タンサク

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Abstract

Freezing of extracellular spaces is the first event when plants are exposed to freezing temperatures, but mechanisms of ice nucleation and propagation remain poorly understood. Japanese cold hardy dwarf bamboos grow in one of the northernmost areas of bamboo distribution and are known to employ deep supercooling in most living tissues as the freeze survival strategy. Studies have been performed on mechanisms of their deep supercooling capability. However, little is known about the ice nucleating factors which may initiate freezing at the apoplast of the vascular and epidermal tissues. This study focused on ice nucleation activity (INA) in aploplast extracts from cold hardy bamboo species, Sasa kurilensis, S. nipponica and Sasamorpha borealis. We obtained apoplast extracts from leaf blades and culms and determined their INA following size fractionation using membrane filters. The apoplast extracts from leaf blades and culms had highly extractable and particulate INA (size: 10 kDa<<0.2 μm). INA of substances larger than 300 kDa (<<0.2 μm) were similar to the freeze initiation temperatures of the vascular and epidermal tissues. Results of heat and protease treatments indicated that the major substance(s) responsible for ice nucleation in the apoplast extracts is proteinaceous, irrespective of the seasons, tissues or species.

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