Jason Liang Pin Ng
  • The Australian National University
研究方向
  • Plant Science
RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings
RT-qPCR法测定拟南芥幼苗RNA稳定性
作者:Tianran Jia and Brandon H. Le日期:07/20/2020,浏览量:5121,Q&A: 0
Steady-state mRNA levels are determined by both the rates of transcription and degradation. Regulation of mRNA stability and/or degradation are key factors that can significantly affect mRNA levels and its biological functions. mRNA stability can be measured indirectly after transcription inhibition. This protocol described a rapid and sensitive method of mRNA stability measurement through quantitative reverse transcription PCR (RT-qPCR) after inhibition of RNA transcription by cordycepin in Arabidopsis seedlings.
Measurement of Arabidopsis thaliana Plant Traits Using the PHENOPSIS Phenotyping Platform
使用PHENOPSIS表型分析平台分析拟南芥植株性状
High-throughput phenotyping of plant traits is a powerful tool to further our understanding of plant growth and its underlying physiological, molecular, and genetic determinisms. This protocol describes the methodology of a standard phenotyping experiment in PHENOPSIS automated platform, which was engineered in INRA-LEPSE (https://www6.montpellier.inra.fr/lepse) and custom-made by Optimalog company. The seminal method was published by Granier et al. (2006). The platform is used to explore and test various ecophysiological hypotheses (Tisné et al., 2010; Baerenfaller et al., 2012; Vile et al., 2012; Bac-Molenaar et al., 2015; Rymaszewski et al., 2017). Here, the focus concerns the preparation and management of experiments, as well as measurements of growth-related traits (e.g., projected rosette area, total leaf area and growth rate), water status-related traits (e.g., leaf dry matter content and relative water content), and plant architecture-related traits (e.g., stomatal density and index and lamina/petiole ratio). Briefly, a completely randomized (block) design is set up in the growth chamber. Next, the substrate is prepared, its initial water content is measured and pots are filled. Seeds are sown onto the soil surface and germinated prior to the experiment. After germination, soil watering and image (visible, infra-red, fluorescence) acquisition are planned by the user and performed by the automaton. Destructive measurements may be performed during the experiment. Data extraction from images and estimation of growth-related trait values involves semi-automated procedures and statistical processing.