Increase in Invaginated Vacuolar Membrane Structure Caused by Plant Cell Expansion by Genotoxic Stress Induced by DNA Double-Strand Breaks

Abstract

Summary Vacuoles occupy 80-90% of a mature plant cell and mainly contribute to all types of cell expansion. Zeocin, an inducer of DNA double-strand breaks, causes cell expansion with endoreduplication. The vacuolar structure after zeocin treatment was examined in tobacco (Nicotiana tabacum) cultured cells expressing GFP fused to a vacuole membrane protein. We found that the genotoxic stress induced the cell expansion with subdivision of the vacuolar lumen by cytoplasmic strands. When a femtosecond laser was used to cut off the cytoplasmic strand, mitochondrial transport along the strand stopped. This suggested that in the elongated cells under the genotoxic stress, the transport of subcellular materials was activated for DNA repair within the damaged cell nucleus by the construction of a network of cytoplasmic strands in the vacuolar lumen.

Auxin decreases chromatin accessibility through the TIR1/AFBs auxin signaling pathway in proliferative cells

“…The tobacco BY-2 cell line was maintained as described previously 54 . The growth conditions and medium were the same for the MM2d cell line and the tobacco BY-2 cell line.…”

“…Drug treatments were applied to tobacco BY-2 cells as described previously 54 . Seven-day-old tobacco BY-2 cells were washed twice with five volumes of 2, 4-D-free LS modified medium 55 and then washed cells (2 mL) were transferred to 95 mL medium containing 0 or 50 µM zeocin, and/or 0.2 mg/mL 2, 4-D.…”

Simplification of vacuole structure during plant cell death triggered by culture filtrates of Erwinia carotovora

“…In mature plant cells, the vacuole constitutes most of a cell's volume, and generally has complex invaginations of its membrane, forming transvacuolar strands, internal sheets, and bulbs (Saito et al ; Uemura et al ; Ruthardt et al ; Higaki et al ; Sheahan et al ; Gattolin et al ; Hashiguchi et al ). These formations of the tonoplast are suggested to have some role in the motility of organelles such as Golgi bodies (Nebenführ et al ), mitochondria (Van Gestel et al ; Hasegawa et al ) or gravity‐sensing amyloplasts (Morita et al ; Hashiguchi et al ). Tonoplast invagination also seems to provide a membrane reservoir for quick cell expansion (Saito et al ; Hicks et al ).…”

2A Peptides Contribute to the Co-Expression of Proteins for Imaging and Genome Editing

“…Genetically-encoded fluorescent proteins conjugated to proteins of interest have innovated the investigation of intracellular localization and expression levels (Shimomura et al 1962, Kato et al 2011, Higashi et al 2013, Hasegawa et al 2014, Nozaki et al 2014, Hirakawa et al 2015, Sato et al 2015, Hirakawa and Matsunaga 2016, Inada 2016, 2017, Kawamoto et al 2016, Mizuta et al 2016, Kurita et al 2017, Okamura et al 2017, Takase and Suzuki 2018, Watanabe-Asaka et al 2018. Additionally, imaging with a combination of proteins connected to different types of fluorescent proteins has provided us with a wide range of novel information in different fields (Tsien 1998, Mizuta et al 2016 However, the co-expression of more than one gene requires time-consuming experimental steps to be carried out, including the repeated retransformation or co-transformations of different vectors with various resistant genes.…”