Protocols in Current Issue
    Detection of Protein S-nitrosothiols (SNOs) in Plant Samples on Diaminofluorescein (DAF) Gels
    Authors:  Marta Rodríguez-Ruiz, Paulo T. Mioto, José M. Palma and Francisco J. Corpas , date: 09/20/2017, view: 87, Q&A: 0
    [Abstract] In plant cells, the analysis of protein S-nitrosothiols (SNOs) under physiological and adverse stress conditions is essential to understand the mechanisms of Nitric oxide (NO)-based signaling. We adapted a previously reported protocol for detecting protein SNOs in animal systems (King et al., 2005) for plant samples. Briefly, ...
    Construction of a Single Transcriptional Unit for Expression of Cas9 and Single-guide RNAs for Genome Editing in Plants
    Authors:  Xu Tang, Zhaohui Zhong, Xuelian Zheng and Yong Zhang, date: 09/05/2017, view: 373, Q&A: 0
    [Abstract] The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein9 (Cas9) is a simple and efficient tool for genome editing in many organisms including plant and crop species. The sgRNAs of the CRISPR/Cas9 system are typically expressed from RNA polymerase III promoters, such as U6 and U3. In many transformation events, ...
    Isolation and Quantification of Plant Extracellular Vesicles
    Authors:  Brian D. Rutter, Katie L. Rutter and Roger W. Innes, date: 09/05/2017, view: 325, Q&A: 0
    [Abstract] Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the ...
    Ubiquitin Proteasome Activity Measurement in Total Plant Extracts
    Authors:  Suayib Üstün and Frederik Börnke, date: 09/05/2017, view: 314, Q&A: 0
    [Abstract] The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to ...
    [Bio101] Leaf Clearing Protocol to Observe Stomata and Other Cells on Leaf Surface
    Author:  Nidhi Sharma, date: 09/05/2017, view: 157, Q&A: 0
    [Abstract] In this protocol, leaves are cleared and fixed in an ethanol and acetic acid solution, and mounted in Hoyer’s solution. The cleared leaves are imaged under differential interference contrast (DIC) microscope. This protocol is beneficial for studying stomata, hair cells, and other epidermal cells in plants.
    Using Silicon Polymer Impression Technique and Scanning Electron Microscopy to Measure Stomatal Aperture, Morphology, and Density
    Authors:  Hui-Chen Wu, Ya-Chen Huang, Chia-Hung Liu and Tsung-Luo Jinn, date: 08/20/2017, view: 409, Q&A: 0
    [Abstract] The number of stomata on leaves can be affected by intrinsic development programming and various environmental factors, in addition the control of stomatal apertures is extremely important for the plant stress response. In response to elevated temperatures, transpiration occurs through the stomatal apertures, allowing the leaf to cool through ...
    Isolation of Cytosol, Microsome, Free Polysomes (FPs) and Membrane-bound Polysomes (MBPs) from Arabidopsis Seedlings
    Authors:  Yonghui Zhao and Shengben Li, date: 08/05/2017, view: 480, Q&A: 0
    [Abstract] The plant endomembrane system plays vital roles for synthesis, modification and secretion of proteins and lipids. From the classic view, only mRNAs encoding secreted proteins could be targeted to the endoplasmic reticulum (ER) for translation via a co-translational translocation manner, however, recently this model has been challenged by ...
    Isolation of Guard-cell Enriched Tissue for RNA Extraction
    Authors:  Pirko Jalakas, Dmitry Yarmolinsky , Hannes Kollist and Mikael Brosche, date: 08/05/2017, view: 483, Q&A: 0
    [Abstract] This is a protocol for isolation of guard cell enriched samples from Arabidopsis thaliana plants for RNA extraction. Leaves are blended in ice-water and filtered through nylon mesh to obtain guard cell enriched fragments. With guard cell enriched samples, gene expression analysis can be done, e.g., comparing different gene ...
    Polyamine and Paraquat Transport Assays in Arabidopsis Seedling and Callus
    Authors:  Haoxi Chai, Yun Shen and Huazhong Shi, date: 08/05/2017, view: 346, Q&A: 0
    [Abstract] Polyamines (PAs) are polycationic compounds found in all living organisms and play crucial roles in growth and survival. We here show the ‘Polyamine and paraquat (PQ) transport assay’ protocol, which can be used to examine the uptake activity of PA/PQ transporters. We have used this protocol to demonstrate that PUT3 in Arabidopsis is a ...
    GUS Staining of Guard Cells to Identify Localised Guard Cell Gene Expression
    Authors:  Zhao Liu, Wei Wang, Chun-Guang Zhang, Jun-Feng Zhao and Yu-Ling Chen , date: 07/20/2017, view: 819, Q&A: 0
    [Abstract] Determination of a gene expression in guard cells is essential for studying stomatal movements. GUS staining is one means of detecting the localization of a gene expression in guard cells. If a gene is specially expressed in guard cells, the whole cotyledons or rosette leaf can be used for GUS staining. However, if a gene is expressed in both ...