Isolation of Intact Vacuoles from Petunia Petals and Extraction of  Sequestered Glycosylated Phenylpropanoid Compounds

Skaliter, Oded; Ravid, Jasmin; Cna’ani, Alon; Dvir, Gony; Knafo, Rafael; Vainstein, Alexander

doi:10.21769/bioprotoc.2912

Cited by 2 publications

(1 citation statement)

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“…The data in 4th column (*) are from a previous study (Manusadžianas et al, 2017 over other methods. In particular, the perfusion method (Sakano & Tazawa, 1984) or isolation of the vacuole from higher plant protoplasts (Robert et al, 2007;Skaliter et al, 2018) are superior methods for isolating a pure, single vacuole from a single mature cell; however, the first one is somewhat time-consuming, while another is also quite expensive. The procedure described in the present study can be conducted on turgorless cells within seconds.…”

Section: Resultsmentioning

confidence: 99%

Biomarker identification of isolated compartments of the cell wall, cytoplasm and vacuole from the internodal cell of characean Nitellopsis obtusa

Gylytė

Jurkonienė

Cimmperman

et al. 2021

PeerJ

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Cells of characean algae are attractive for plant cell physiologists because of their large size and their close relation to higher plant cells. The objective of our study was to evaluate the purity of the compartments (cell wall, cytoplasm with plastids, mitochondria, nuclei and endomembrane system, and vacuole) separated mechanically from the internodal cells of Nitellopsis obtusa using enzymatic markers. These included α-mannosidase and malate dehydrogenase, vacuolar and cytoplasmic enzymes, respectively. The biomarkers applied revealed the degree of compartment contamination with the material from unwanted cell parts. The cell wall was contaminated slightly by vacuole and cytoplasm residuals, respectively by 12.3 and 1.96% of corresponding biomarker activities. Relatively high activity of vacuolar marker in the cell wall could be associated with the cell vacuoles in the multicellular structure of the nodes. The biomarkers confirmed highly purified vacuolar (99.5%) and cytoplasmic (86.7%) compartments. Purity estimation of the cell fractions enabled reevaluating nCuO related Cu concentrations in the compartments of charophyte cell. The internalisation of CuO nanoparticles in N. obtusa cell occurred already after 0.5h. In general, the approach seems to be useful for assessing the accumulation and distribution of various xenobiotics and/or metabolites within plant cell. All this justifies N.obtusa internodal cells as a model organism for modern studies in cell biology and nanotoxicology.

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