Hydrogen peroxide production is an early event during bicarbonate induced stomatal closure in abaxial epidermis of Arabidopsis

Abstract

The presence of 2 mM bicarbonate in the incubation medium induced stomatal closure in abaxial epidermis of Arabidopsis. Exposure to 2 mM bicarbonate elevated the levels of H(2)O(2) in guard cells within 5 min, as indicated by the fluorescent probe, dichlorofluorescein diacetate (H(2)DCF-DA). Bicarbonate-induced stomatal closure as well as H(2)O(2) production were restricted by exogenous catalase or diphenylene iodonium (DPI, an inhibitor of NAD(P)H oxidase). The reduced sensitivity of stomata to bicarbonate and H(2)O(2) production in homozygous atrbohD/F double mutant of Arabidopsis confirmed that NADP(H) oxidase is involved during bicarbonate induced ROS production in guard cells. The production of H(2)O(2) was quicker and greater with ABA than that with bicarbonate. Such pattern of H(2)O(2) production may be one of the reasons for ABA being more effective than bicarbonate, in promoting stomatal closure. Our results demonstrate that H(2)O(2) is an essential secondary messenger during bicarbonate induced stomatal closure in Arabidopsis.

Increased protein carbonylation in leaves of Arabidopsis and soybean in response to elevated [CO2]

“…Our results are also consistent with recent studies that suggested that elevated [CO 2 ] can increase ROS production in vivo. For example, the bicarbonate-induced stomatal closure in epidermal peels of Arabidopsis was shown to be the result of increased H 2 O 2 production in guard cells (Kolla et al 2006;Kolla et al 2007). Likewise, Cheeseman (2006) found that leaves of soybean plants grown at elevated [CO 2 ] had higher levels of H 2 O 2 compared to plants at ambient [CO 2 ], although measurement of tissue H 2 O 2 content can be difficult and problematic (Queval et al 2008).…”

Reactive oxygen species and plant resistance to fungal pathogens

“…They consist in non-fluorescent molecules that become fluorescent when oxidised by ROS, and the emitted fluorescence can be observed by fluorimetry and/or by fluorescent microscopy, an advantage of such probes (Benikhlef et al, 2013;Bulgakov et al, 2012;Fester and Hause, 2005;Guo et al, 2010;Kolla et al, 2007;L'Haridon et al, 2011;Li et al, 2007;Liu et al, 2010;Ma et al, 2013;Peleg-Grossman et al, 2012;Plancot et al, 2013;Tada et al, 2004;Wen et al, 2008;Ye et al, 2013). Luminol or luminol analogues are sensitive chemiluminescent probes used to quantify a relative intensity of ROS by counting the emitted light with a luminometer, CDD camera or a scintillation counter (Dubreuil-Maurizi et al, 2010;Flury et al, 2013;Kunz et al, 2006;L'Haridon et al, 2011;Mersmann et al, 2010).…”

Functional Convergence of Oxylipin and Abscisic Acid Pathways Controls Stomatal Closure in Response to Drought

“…Abaxial (lower) epidermal strips of Arabidopsis, tomato (Solanum lycopersicum), and Brassica napus were adhered to glass cover slips using Telesis V adhesive (Premiere Products) and subsequently treated as previously described (Kolla et al, 2007). The epidermal strips incubated with various phytohormones were irradiated with white light for 2 h. The width of the stomatal aperture was measured by light microscope (Nikon OPTIPHOT 2) using a precalibrated ocular micrometer.…”