Plant Science

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    Protocols in Current Issue
    Enrichment of Cytoplasmic RNA Granules from Arabidopsis Seedlings
    Authors:  Zhen Lei, Eun Yu Kim and Jungnam Cho, date: 11/05/2021, view: 1618, Q&A: 0
    [Abstract]

    RNA granules (RGs) are membraneless intracellular compartments that play important roles in the post-transcriptional control of gene expression. Stress granules (SGs) are a type of RGs that form under environmental challenges and/or internal cellular stresses. Stress treatments lead to strong mRNAs translational inhibition and storage in SGs until

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    Quantitative Analysis of RNA Editing at Specific Sites in Plant Mitochondria or Chloroplasts Using DNA Sequencing
    Authors:  Yang Yang and Weixing Shan, date: 09/20/2021, view: 1536, Q&A: 0
    [Abstract]

    Cytidine-to-uridine (C-to-U) RNA editing is one of the most important post-transcriptional RNA processing in plant mitochondria and chloroplasts. Several techniques have been developed to detect the RNA editing efficiency in plant mitochondria and chloroplasts, such as poisoned primer extension (PPE) assays, high-resolution melting (HRM) analysis,

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    U2.3 Precursor Small Nuclear RNA in vitro Processing Assay
    Authors:  Chan Lin, Yujie Feng, Xueyan Peng, Jiaming Wu, Weili Wang and Yunfeng Liu, date: 09/05/2021, view: 1173, Q&A: 0
    [Abstract]

    Small nuclear RNAs (snRNAs) are vital for eukaryotic cell activities and play important roles in pre-mRNA splicing. The molecular mechanism underlying the transcription of snRNA, regulated via upstream/downstream cis-elements and relevant trans-elements, has been investigated in detail using cell-free extracts. However, the processing of precursor

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    A Novel Method to Map Small RNAs with High Resolution
    Authors:  Kun Huang, Feray Demirci, Blake C. Meyers and Jeffrey L. Caplan, date: 08/20/2021, view: 2205, Q&A: 0
    [Abstract]

    Analyzing cellular structures and the relative location of molecules is essential for addressing biological questions. Super-resolution microscopy techniques that bypass the light diffraction limit have become increasingly popular to study cellular molecule dynamics in situ. However, the application of super-resolution imaging techniques to detect

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    Micrografting in Arabidopsis Using a Silicone Chip
    Authors:  Hiroki Tsutsui, Yaichi Kawakatsu and Michitaka Notaguchi, date: 06/20/2021, view: 3845, Q&A: 1
    [Abstract]

    The micrografting technique in the model plant Arabidopsis has been widely used in the field of plant science. Grafting experiments have demonstrated that signal transductions are systematically regulated in many plant characteristics, including defense mechanisms and responses to surrounding environments such as soil and light conditions.

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    Monitoring Real-time Temperature Dynamics of a Short RNA Hairpin Using Förster Resonance Energy Transfer and Circular Dichroism
    Authors:  Martin Balcerowicz, Marco Di Antonio and Betty Y. W. Chung, date: 03/20/2021, view: 3451, Q&A: 0
    [Abstract]

    RNA secondary structures are highly dynamic and subject to prompt changes in response to the environment. Temperature in particular has a strong impact on RNA structural conformation, and temperature-sensitive RNA hairpin structures have been exploited by multiple organisms to modify the rate of translation in response to temperature changes.

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    Plant ARGONAUTE Protein Immunopurification for Pathogen Cross Kingdom Small RNA Analysis
    Authors:  Florian Dunker, Bernhard Lederer and Arne Weiberg, date: 02/05/2021, view: 3603, Q&A: 0
    [Abstract]

    Over the last decade, it has been noticed that microbial pathogens and pests deliver small RNA (sRNA) effectors into their host plants to manipulate plant physiology and immunity for infection, known as cross kingdom RNA interference. In this process, fungal and oomycete parasite sRNAs hijack the plant ARGONAUTE (AGO)/RNA-induced silencing complex

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    RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings
    Authors:  Tianran Jia and Brandon H. Le, date: 07/20/2020, view: 3574, Q&A: 0
    [Abstract] 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 ...
    Real-time PCR Analysis of PAMP-induced Marker Gene Expression in Nicotiana benthamiana
    Authors:  Fan Liu, Yuanpeng Xu, Yan Wang and Yuanchao Wang, date: 10/05/2018, view: 6993, Q&A: 0
    [Abstract] Perception of pathogen-associated molecular patterns (PAMPs) often triggers various innate immune responses in plants. The transcriptional changes of defense-related genes are often used as a marker to assay PAMP-triggered plant immune response. Here we described a protocol to monitor the relative expression level of marker genes in Nicotiana ...
    In planta Transcriptome Analysis of Pseudomonas syringae
    Authors:  Tatsuya Nobori and Kenichi Tsuda, date: 09/05/2018, view: 7355, Q&A: 0
    [Abstract] Profiling bacterial transcriptome in planta is challenging due to the low abundance of bacterial RNA in infected plant tissues. Here, we describe a protocol to profile transcriptome of a foliar bacterial pathogen, Pseudomonas syringae pv. tomato DC3000, in the leaves of Arabidopsis thaliana at an early stage of ...



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