宽频耗散型石英晶体微天平实时监测PEG6000胁迫下烟草细胞的粘弹性响应  

Real-time Monitoring of Viscoelastic Responses of Tobacco(Nicotiana tabacum)Cells Under PEG6000 Stress by Wide Frequency Quartz Crystal Microbalance with Dissipation

作　　者：张煜 周铁安[1,2] 陈宗星 潘炜松[1,2] 谭成方 ZHANG Yu;ZHOU Tie-An;CHEN Zong-Xing;PAN Wei-Song;TAN Cheng-Fang(College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha 410128,China;Hunan Provincial Engineering Technology Research Center for Cell Mechanics and Function Analysis,Changsha 410128,China)

机构地区：[1]湖南农业大学生物科学技术学院,长沙410128 [2]湖南省细胞力学与功能分析工程技术研究中心,长沙410128

出　　处：《农业生物技术学报》2020年第12期2270-2280,共11页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31870933);湖南省自然科学基金面上项目(2018JJ2164)。

摘　　要：作物产量由其遗传与抗逆性决定,现今抗逆性育种主要以植株为受试对象进行种植,受环境制约大、试验周期长。为寻找一种便捷且科学的植物抗逆评估方法,本研究以活细胞表征植株的方法为策略,以聚二烯丙基二甲基氯化铵(poly dimethyl diallyl ammonium chloride,PDADMAC)为静电吸附剂,将烟草(Nicotiana tabacum)BY-2细胞黏附在耗散型石英晶体微天平Q-sense的5 MHz晶体金电极表面,并通过Q-sense实时监测BY-2细胞在5%~20%PEG6000胁迫下的粘弹性响应。结果表明,PDADMAC可作为烟草BY-2细胞黏附剂,细胞在黏附过程中耗散增加、频率下降;PEG6000胁迫下细胞均变软,且细胞骨架比细胞壁更软。细胞在接触PEG6000的瞬间胁迫现象最明显,胁迫程度受到PEG6000浓度与处理时间的影响:在5%PEG6000胁迫下,细胞在瞬态变软后持续变硬;在更高浓度PEG6000胁迫下,细胞在瞬态变软后仅瞬态变硬,然后稍有继续变硬(15%PEG6000)或不再变硬(20%PEG6000)。此外,应用Q-sense技术比较PEG6000在0%~20%顺浓度梯度及之后20%~0%逆浓度梯度变化的细胞响应规律,总的趋势是顺浓度胁迫下细胞逐渐变软、逆浓度处理下细胞逐渐变硬。本研究实时、定量评估了PEG6000胁迫对烟草BY-2细胞粘弹性的影响,为研究植物干旱胁迫提供了新的方法,有助于细胞层面的抗逆种质鉴别与评估。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 wa

关 键 词：石英晶体微天平 烟草BY-2细胞 PEG6000 干旱胁迫 细胞粘弹性指数 

分 类 号：S313[农业科学—作物栽培与耕作技术]