生物发光成像无损伤研究植物生物钟的方法

doi:10.11983/CBB19238

摘要/Abstract

摘要： 植物生物钟系统是植物为了适应地球自转进化出的以约24小时为周期的分子系统, 通过感知并整合外界周期性变化的环境信号进而协调细胞内相应基因的表达和能量状态, 赋予植物对生存环境的适应性并参与调控多个植物生长发育过程。目前, 越来越多的研究聚焦于解析植物生物钟的分子机制, 基于此也衍生出很多研究生物钟表型的方法。该文在总结已有生物钟检测方法的基础上, 重点介绍生物钟表型实验中最常用且比较稳定可靠的实验方法, 以期为生物钟的表型研究尤其是生物钟机制研究提供技术支持与借鉴。

关键词: 生物钟, 表型分析, FFT-NLLS, BRASS

Abstract: The plant circadian clock is a time-keeping molecular system, with a cycle of ~ 24 h. It was evolved to adapt to the diel rhythmic environmental cues generated by the self-rotation of the Earth. In addition to the time-keeping function, the circadian clock also regulates a plethora of plant growth and development processes by synchronizing the endogenous energy and metabolomic status. By sensing and integrating the dynamics of external environmental cues, circadian clock can coordinate gene expression at multiple levels, thus to increase the fitness of plants. Recently, there is an increasingly demand for measuring and assessing circadian phenotype for many non-circadian research field. Here, we summarized the currently available methods for detecting circadian rhythm, and show one most commonly used standard procedure for evaluating circadian phenotype in plants, which may help to provide the applicable technical assistance for the study of the circadian clock.

Key words: circadian clock, phenotype analysis, FFT-NLLS, BRASS

于英俊,徐航,王雷. 生物发光成像无损伤研究植物生物钟的方法. 植物学报, 2020, 55(2): 177-181.

Yingjun Yu,Hang Xu,Lei Wang. A Non-invasive Method for Measuring and Analyzing Circadian Phenotype in Living Plants. Chinese Bulletin of Botany, 2020, 55(2): 177-181.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB19238

https://www.chinbullbotany.com/CN/Y2020/V55/I2/177

图/表 1

图1 Col-0野生型拟南芥CCA1pro:LUC在持续红光条件下的生物节律分析 (A) 携带有CCA1pro:LUC报告基因的Col-0野生型拟南芥在持续红光条件下, 从LL24到LL144时间范围内生物发光的荧光节律(c.p.s代表每株植物每秒钟采集到的光子数)。(B) 野生型拟南芥生物钟周期的散点图, X轴表示生物钟周期, Y轴表示相对振幅误差; 根据结果显示每株幼苗周期较为集中, 相对一致, 而且相对振幅误差低于0.5, 结果可靠(一般认为当RAE值大于0.5时, 节律的振荡强度偏低, 不适于计算生物钟周期)。(C) 野生型拟南芥生物钟周期的柱状图; 根据结果显示周期长度在持续红光下为23.22小时。计算方法: FFT-NLLS。数据为21株幼苗的平均值±标准误。

Figure 1 Circadian phenotype analysis of Col-0 wild type Arabidopsis thaliana CCA1pro:LUC under continuous red light (A) Trace plot of bioluminescence in Col-0 wild type Arabidopsis thaliana with CCA1pro:LUC reporter under continuous red light conditions, from LL24 to LL144 (the abbreviation of c.p.s represents the counts of photons per seedling). (B) Scatter plot of circadian period (x-axis) and relative amplitude error (RAE) (y-axis) of Col-0; According to the results, the periods are more concentrated and relatively consistent, and the relative amplitude errors are all lower than 0.5, which indicates that the result is reliable (generally, when the RAE is greater than 0.5, the circadian oscillation intensity is relatively low, which is not suitable for calculating the period). (C) Estimated circadian period of CCA1pro:LUC in Col-0; According to the calculation, circadian period length of Col-0 is 23.22 hours under continuous red light. Calculation was performed using FFT-NLLS method. Data are presented as means±SE (n=21).

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