Monitoring the dynamics of starch in guard cells

 

 

Quantification of Starch in Guard Cells of Arabidopsis thaliana 

by Flütsch S., Distefano L., Santelia D. (2018)

Department of Plant and Microbial Biology, University of Zürich, Zürich, Switzerland

Sabrina Flütsch, Luca Distefano, Diana Santelia,

20180528042414448Santelia Diana portrait2013
Diana Santelia, Department of Plant and Microbial Biology, University of Zürich, Zürich, Switzerland

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in Bio-protocol 8, Iss 13 – DOI:   10.21769/BioProtoc.2920

https://bio-protocol.org/e2920

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.

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A method of preparing enriched stomatal guard cells

 

 

Stomata Tape-Peel: An Improved Method for Guard Cell Sample Preparation

by Lawrence S., Pang Q., Kong W., Chen S. (2018)

Sheldon Lawrence II1,2,3, Quiying Pang2,3,4, Wenwen Kong2,3, Sixue Chen1,2,3,5
Plant Molecular and Cellular Biology Program, University of Florida,
Department of Biology, University of Florida,
Genetics Institute (UFGI), University of Florida,
Alkali Soil Natural Environmental Science Center, Northeast Forestry University,
Interdisciplinary Center for Biotechnology Research (ICBR), University of Florida

 

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in Jove 137 – DOI: 10.3791/57422 –

https://www.jove.com/video/57422/stomata-tape-peel-an-improved-method-for-guard-cell-sample-preparation

SUMMARY

This protocol describes a method of preparing enriched stomatal guard cells that is useful for physiological and other biological studies.

Counting stomata

Screen Shot 2018-09-27 at 10.01.23

 

Counting stomata from different types of leaves

by Suthar K., Sutariya K., Yadav P., Menon S. (2017)

Kirti Suthjar, Komal Sutariya, Poola yadav, Sheeba Menon,

Biology Department, K. K. Shah Jarodwala Maninagar Science College, Maninagar, Almedabad

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in Int. J. Trend in Scientific Res. & Dev. (IJTSRD) 1(6): 1068-1075 – ISSN 2456-6470 –

https://www.academia.edu/35945205/Counting_of_Stomata_from_Different_Types_of_Leaves

Screen Shot 2018-09-27 at 09.54.35
 

Method for studying grass leaf epidermis and stomata

 

 

Convenient method for studying grass leaf epidermis

by Hilu K. W., Randall J. L. (1984)

Khidir_Hilu
Khidir Hilu,, Virginia Polytechnic Institute and State University, Blacksburg, United States

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in Taxon 33(3): 413- 415 – DOI: 10.2307/1220980 –

https://www.jstor.org/stable/1220980?seq=1#page_scan_tab_contents

Abstract

Leaf epidermis of grasses is structurally elaborate and quite important in modern systematics of the Poaceae.
Available techniques used in preparing leaf epidermis for microscopic studies are time-consuming and do not always produce adequate preparations.
A simple and convenient technique is proposed in this paper which requires the application of a film of clear nail polish directly to the leaf surface.
The impressions left on the polish film after drying produce an excellent detailed image of the epidermis. This technique is also applicable to other plant families.

Functionally isolated guard cells (stomata) in epidermal strips

 

 

A simple technique of obtaining functionally isolated guard cells in epidermal strips of Vicia faba

by Durbin R. D., Graniti A. (1975)

Richard D. Durbin, Antonio Graniti,

Centro di studio del CNR su le tossine e i parassiti sistemici dei vegetali Istituto di Patologia vegetale, Università:Bari, Italy

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in Planta 126: 285-288 – doi: 10.1007/BF00388970 –

https://www.ncbi.nlm.nih.gov/pubmed/24430222

Abstract

Ultrasonic irradiation of Vicia faba L. epidermal strips for 2 min preferentially disrupts the epidermal cells but does not impair guard cell movements.

Maximal opening induced by fusicoccin requires that K(+) be provided to the guard cells from external sources. A mobile organic anion is not required.

Introduction

Procedures for obtaining epidermal strips with only the guard cells alive have been developed but they are either tedious, require considerable experience or may damage the guard cells (Meidner and Mansfield, 1968; Squire and Mansfield, 1972b; Allaway and Hsiao, 1973). Such material is, however, very useful for studies on some aspects of the mechanism of guard cell movements. In this paper a simple, rapid and reproducible technique using ultrasonic irradiation to preferentially disrupt epidermal cells is reported which largely overcomes these drawbacks. ll[aterials

Preparation of leaf epidermis for experiments on stomatal physiology

 

 

Selection and preparation of leaf epidermis for experiments on stomatal physiology

by Weyers J. D. B., Travis A. J. (1981)

Screen Shot 2018-07-17 at 20.51.10

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in J. Exp. Bot. 32, 837–850 – doi: 10.1093/jxb/32.4.837 –

https://www.jstor.org/stable/23690065?seq=1#page_scan_tab_contents

Abstract

A simple method for reproducibly peeling leaf epidermis tissue is described. Epidermis was obtained from eight species using the method, and its properties of value in research on stomatal physiology evaluated.
Effects of the method of peeling on cell viability and mesophyll contamination were quantified, and a comparison was made between the effects of peeling and subsequent treatments on a plant with morphologically distinct subsidiary cells, Commelina communis, and one without, Vicia faba.
The results indicate that artefacts in experiments on stomatal physiology involving leaf epidermis could arise not only from the peeling method, but also the plant species chosen.

Stomata and epidermal cells of herbaria specimens

 

 

Non-invasive method for looking at stomata / epidermal cells of herbaria specimens

by Wyatt A. (2018)

Amy_Wyatt5
Amy Wyatt, Cardiff University, UK

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in Project DOI: 10.13140/RG.2.2.15999.30884 –

https://www.researchgate.net/publication/324784278_Non-invasive_method_for_looking_at_stomata_epidermal_cells_of_herbaria_specimens

Earlier on in the year I perfected a non-invasive method to look at stomatal density and guard cell size of herbarium specimens. After preliminary tests using clear nail varnish proved unsuccessful, I found a method online (http://www.saps.org.uk/secondary/teaching-resources/299-measuring-stomatal-density-) that involved using Germolene liquid plaster on the herbaria specimens to make an imprint of the leaf surface. For each individual, a photo was taken marking leaves 1-5 with a number so later on I know which leaves have been analysed, and I can account for leaf area, length and width when determining the stomatal density / guard cell size of individuals.

METHOD:

1.) Spread germolene, liquid skin over surface of leaf in a 1cm2 patch and allow to dry for 5-10 minutes so it is no longer tacky on the leaf.

2.) The germolene will dry transparent, and you will need to gently pry the surface of the germolene until it goes opaque. I was using dental tools for this, the ‘dental explorer’ tool (pictured below) with a right-angled head was perfect for this. Run the outside edge of right angle (not the sharp end) gently over the surface of the germolene, you will start to see the germolene turning opaque as it detached from the leaf surface.

3.) Once the whole surface has turned opaque, use the sharp end of the tool to gently pry at the edge of the germolene until it starts to peel off, use fine tweezers to remove gently without damaging the leaf.

4.) Once you have your germolene imprint, mount onto a microscope slide and place a cover slip over the top of the sample.

*** for the species I was working with Impatiens glandulifera I took one measurement from the left hand side of the abaxial surface 1cm from the leaf vein.