Real-time Monitoring of Viscoelastic Responses of Tobacco (Nicotiana tabacum) Cells Under PEG6000 Stress by Wide Frequency Quartz Crystal Microbalance with Dissipation
ZHANG Yu1,2, ZHOU Tie-An1,2,*, CHEN Zong-Xing1,2, PAN Wei-Song1,2, TAN Cheng-Fang1,2
1 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China; 2 Hunan Provincial Engineering Technology Research Center for Cell Mechanics and Function Analysis, Changsha 410128, China
Abstract：Crop yield is determined by its genetics and stress resistance. Currently stress resistance breeding mainly uses plants as test objects, which is subject to environmental constraints and long test cycles. In order to find a convenient and scientific method for assessing plant stress resistance, this study took the strategy of using living cells to characterize plant. Tobacco (Nicotiana tabacum) BY-2 cells were adhered to the gold electrode surface of 5 MHz Quartz Crystal Microbalance with Dissipation (Q-sense) by using poly dimethyl diallyl ammonium chloride (PDADMAC) as electrostatic adsorbent, and the viscoelastic responses of BY-2 cells under stresses of 5%~20% PEG6000 were monitored in real time by Q-sense. The results showed that PDADMAC could be used as an adhesion agent for tobacco BY-2 cells, the Q-sense's dissipation increased and frequency decreased during the adhesion of BY-2 cells; the cells became softer under the stresses of PEG6000, and the cytoskeleton was softer than the cell wall. The Q-sense's responses were the most obvious at the moments when the cells were exposed to PEG6000, which was also affected by the PEG6000 concentration and the treatment time. Under the stress of 5% PEG6000, the cells continued to harden after a transient softening; while under the stresses of 15% and 20% PEG6000, the cells only experienced a brief hardening after the transient softening followed by a slightly further hardening (15% PEG6000) or no longer hardening (20% PEG6000). In addition, the Q-sense technique was used to compare the responses of BY-2 cells subjected to the treatments of 0%~20% PEG6000 concentration gradient and the subsequent 20%~0% PEG6000 inverse concentration gradient, the general trend was that the cells became gradually softer under the stresses of normal concentration gradient then the cells became gradually harder under the treatment of reverse concentration gradient. This study assessed the effect of PEG6000 stress on the viscoelastic changes of tobacco BY-2 cells in a real-time and quantitative manner, providing a new way for studying plant drought stress which would be helpful for the identification and evaluation of resistance germplasm at the cellular level.
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