Nitric oxide (NO) measurements in stomatal guard cells

dc.contributor.author Agurla, Srinivas
dc.contributor.author Gayatri, Gunja
dc.contributor.author Raghavendra, Agepati S.
dc.date.accessioned 2022-03-27T03:50:28Z
dc.date.available 2022-03-27T03:50:28Z
dc.date.issued 2016-04-01
dc.description.abstract The quantitative measurement of nitric oxide (NO) in plant cells acquired great importance, in view of the multifaceted function and involvement of NO as a signal in various plant processes. Monitoring of NO in guard cells is quite simple because of the large size of guard cells and ease of observing the detached epidermis under microscope. Stomatal guard cells therefore provide an excellent model system to study the components of signal transduction. The levels and functions of NO in relation to stomatal closure can be monitored, with the help of an inverted fluorescence or confocal microscope. We can measure the NO in guard cells by using flouroprobes like 4,5-diamino fluorescein diacetate (DAF-2DA). This fluorescent dye, DAF-2DA, is cell permeable and after entry into the cell, the diacetate group is removed by the cellular esterases. The resulting DAF-2 form is membrane impermeable and reacts with NO to generate the highly fluorescent triazole (DAF-2T), with excitation and emission wavelengths of 488 and 530 nm, respectively. If time-course measurements are needed, the epidermis can be adhered to a cover-glass or glass slide and left in a small petri dishes. Fluorescence can then be monitored at required time intervals; with a precaution that excitation is done minimally, only when a fluorescent image is acquired. The present method description is for the epidermis of Arabidopsis thaliana and Pisum sativum and should work with most of the other dicotyledonous plants.
dc.identifier.citation Methods in Molecular Biology. v.1424
dc.identifier.issn 10643745
dc.identifier.uri 10.1007/978-1-4939-3600-7_5
dc.identifier.uri http://link.springer.com/10.1007/978-1-4939-3600-7_5
dc.identifier.uri https://dspace.uohyd.ac.in/handle/1/5717
dc.subject 4,5-Diamino fluorescein diacetate
dc.subject Nitric oxide
dc.subject Reactive nitrogen species
dc.subject Stomatal guard cells
dc.subject Time-course measurement
dc.title Nitric oxide (NO) measurements in stomatal guard cells
dc.type Book Series. Book Chapter
dspace.entity.type
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