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    Protocols in Current Issue
    High Resolution Melting Temperature Analysis to Identify CRISPR/Cas9 Mutants from Arabidopsis
    Authors:  Cynthia Denbow, Sonia Carole Ehivet and Sakiko Okumoto, date: 07/20/2018, view: 26, Q&A: 0
    CRISPR/Cas9 made targeted mutagenesis and genome editing possible for many plant species. One of the ways that the endonuclease is used for plant genetics is the creation of loss-of-function mutants, which typically result from erroneous DNA repair ...
    Extraction of RNA from Recalcitrant Tree Species Paulownia elongata
    Authors:  Niveditha Ramadoss and Chhandak Basu, date: 07/20/2018, view: 34, Q&A: 0
    Isolation of pure RNA is the basic requisite for most molecular biology work. Plants contain polyphenols and polysaccharides, which can interfere with isolation of pure RNA from them. Especially hardwood tree species like Paulownia elongata ...
    High Resolution Melting Temperature Analysis to Identify CRISPR/Cas9 Mutants from Arabidopsis
    Authors:  Cynthia Denbow, Sonia Carole Ehivet and Sakiko Okumoto, date: 07/20/2018, view: 26, Q&A: 0
    [Abstract] CRISPR/Cas9 made targeted mutagenesis and genome editing possible for many plant species. One of the ways that the endonuclease is used for plant genetics is the creation of loss-of-function mutants, which typically result from erroneous DNA repair through non-homologous end joining (NHEJ) pathway. The majority of erroneous repair events results ...
    Extraction of RNA from Recalcitrant Tree Species Paulownia elongata
    Authors:  Niveditha Ramadoss and Chhandak Basu, date: 07/20/2018, view: 34, Q&A: 0
    [Abstract] Isolation of pure RNA is the basic requisite for most molecular biology work. Plants contain polyphenols and polysaccharides, which can interfere with isolation of pure RNA from them. Especially hardwood tree species like Paulownia elongata have surplus amount of RNA-binding alkaloids, proteins and secondary metabolites that can further ...
    Quantification of Starch in Guard Cells of Arabidopsis thaliana
    Authors:  Sabrina Flütsch, Luca Distefano and Diana Santelia, date: 07/05/2018, view: 233, Q&A: 0
    [Abstract] In this protocol, we describe how to quantify starch in guard cells of Arabidopsis thaliana using the fluorophore propidium iodide and confocal laser scanning microscopy. This simple method enables monitoring, with unprecedented resolution, the dynamics of starch in guard cells.
    Separation of Thylakoid Protein Complexes with Two-dimensional Native-PAGE
    Authors:  Marjaana Rantala, Virpi Paakkarinen and Eva-Mari Aro, date: 07/05/2018, view: 282, Q&A: 0
    [Abstract] The hierarchical composition and interactions of the labile thylakoid protein complexes can be assessed by sequential 2D-native gel-electrophoresis system. Mild non-ionic detergent digitonin is used to solubilize labile protein super-and megacomplexes, which are then separated with first-dimension blue native polyacrylamide gel electrophoresis ...
    A Modified Approach for Axenic Cultivation of Spores of Fern Adiantum capillus-veneris L. with High Germination Rate
    Authors:  Yuan Ya-Ning, Yi Shu, Jin Wan-Ting and Fang Yu-Han, date: 07/05/2018, view: 160, Q&A: 0
    [Abstract] Spores are the primary way of spread and reproduction for ferns, a clade of seed-free vascular plants. However, no detailed protocol for ferns spore cultivation has been reported yet. Here we provide a modified approach for axenic cultivation of fern Adiantum capillus-veneris L., based on Cao’s and Li’s method (Cao, et al., 2010; ...
    Isolation of Intact Vacuoles from Petunia Petals and Extraction of Sequestered Glycosylated Phenylpropanoid Compounds
    Authors:  Oded Skaliter , Jasmin Ravid, Alon Cna'ani, Gony Dvir, Rafael Knafo and Alexander Vainstein, date: 07/05/2018, view: 146, Q&A: 0
    [Abstract] Plant vacuoles are the largest compartment in plant cells, occupying more than 80% of the cell volume. A variety of proteins, sugars, pigments and other metabolites are stored in these organelles (Paris et al., 1996; Olbrich et al., 2007). Flowers produce a variety of specialized metabolites, some of which are unique to this ...
    An Optimized CTAB Method for Genomic DNA Extraction from Freshly-picked Pinnae of Fern, Adiantum capillus-veneris L.
    Authors:  Yi Shu, Jin Wan-Ting, Yuan Ya-Ning and Fang Yu-Han, date: 07/05/2018, view: 172, Q&A: 0
    [Abstract] As the sister clade of seed plants, ferns are significant materials for plant phylogeny research. However, the genomic DNA extraction protocol for fern samples like modified CTAB method still lacks robustness. Here, we found that the amount and condition of the pinnae samples are critical for gDNA extraction in fern, Adiantum capillus-veneris ...
    Extraction and 16S rRNA Sequence Analysis of Microbiomes Associated with Rice Roots
    Authors:  Joseph Edwards, Christian Santos-Medellín and Venkatesan Sundaresan, date: 06/20/2018, view: 454, Q&A: 0
    [Abstract] Plant roots associate with a wide diversity of bacteria and archaea across the root-soil spectrum. The rhizosphere microbiota, the communities of microbes in the soil adjacent to the root, can contain up to 10 billion bacterial cells per gram of soil (Raynaud and Nunan, 2014) and can play important roles for the fitness of the host plant. Subsets ...
    Increasing the Membrane Permeability of a Fern with DMSO
    Author:  Marcelo Garcés, date: 06/20/2018, view: 264, Q&A: 0
    [Abstract] Cell membrane prevents the entrance of extra molecules (e.g., transcription and translation inhibitors) into the cell. For studying the physiological effects of transcription and translation inhibitors on Hymenophyllum caudiculatum fronds, we incubate fronds with 0.1% DMSO to test if this increases cell membrane permeability ...
    Analysis of Autophagic Activity Using ATG8 Lipidation Assay in Arabidopsis thaliana
    Authors:  Mengqian Luo and Xiaohong Zhuang, date: 06/20/2018, view: 340, Q&A: 0
    [Abstract] As a fundamental metabolic pathway to degrade and recycle cellular cargos, autophagy is highly induced upon stress, starvation and senescence conditions in plants. A double-membrane structure named autophagosome will form during this process for cargo sequestration and delivery into the vacuole.

    A number of regulators have been ...