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Background: In the last decades, replicating expression vectors based on plant geminivirus have been widely used for enhancing the efficiency of plant transient expression. By using the replicating expression vector derived from bean yellow dwarf virus and green fluorescent protein as a reporter, we investigated the effects of α-naphthalene acetic acid, gibberellins3, and 6-benzyladenine, as three common plant growth regulators, on the plant biomass and efficiency of transient expression during the process of transient expression in Nicotiana benthamiana L. leaves. Results: With the increase of the concentration of α-naphthalene acetic acid, gibberellins3, and 6-benzyladenine (from 0.1 to 1.6 mg/L), the fresh weight, dry weight, and leaf area of the seedlings increased first and then returned to the levels similar to the controls (without chemical treatment). The treatment with α-naphthalene acetic acid at 0.2 and 0.4 mg/L can enhance the level of transient expression of green fluorescent protein, which peaked at 0.4 mg/L α-naphthalene acetic acid and was increased about by 19%, compared to the controls. Gibberellins3 at 0.1–0.4 mg/L can enhance the level of transient expression of green fluorescent protein, which peaked at 0.2 mg/L gibberellins3 and was increased by 25%. However, the application of 6-benzyladenine led to decrease in the level of transient expression of green fluorescent protein. Conclusions: The appropriate plant growth regulators at moderate concentration could be beneficial to the expression of foreign genes from the Agrobacterium-mediated transient expression system in plants. Thus, appropriate plant growth regulators could be considered as exogenous components that are applied for the production of recombinant protein by plant-based transient expression systems. [ABSTRACT FROM AUTHOR]

Extracellular adenosine 5′-triphosphate (ATP) is a well-known signalling molecule in plants and plays important roles during plant stress response. In the present work, the treatment of tobacco cell suspension culture with exogenous hydrogen peroxide (H2O2) caused the decreases of respiratory O2 uptake, intracellular ATP production, extracellular ATP level, and reduction of cell viability. Combining this observation with the finding that the oxidative phosphorylation uncoupler or the respiratory inhibitor not only decreased intracellular ATP production but also decreased the extracellular ATP level and cell viability, it is suggested that the decrease of extracellular ATP level and cell viability under H2O2 stress could be a result of the suppressed production of intracellular ATP. Treatment with ATP-degrading enzyme, apyrase, also caused the decreases of cell viability, respiratory O2 uptake, and intracellular ATP production. More importantly, addition of exogenous ATP alleviated the H2O2-induced decreases of cell viability, respiratory O2 uptake, and production of intracellular ATP. These results indicate that extracellular ATP could be an important effector in regulating the cell viability, respiratory O2 uptake, and intracellular ATP production of tobacco cell suspension culture under H2O2 stress. [ABSTRACT FROM AUTHOR]

In the present work, by using tobacco cell suspension and wheat seedlings, we studied that eATP (extracellular ATP) released by copper (Cu) stress could act as diffusible signal in alleviating the Cu stress-induced cell death. A semipermeable membrane was fixed in the middle of a plastic box to divide the box into two equal compartments (A and B, respectively). This semipermeable membrane can prevent direct cell-to-cell (or seedling-to-seedling) contact and the diffusion of the macromolecules [such as ATPase (adenosine 5′-triphosphatase)] between these two compartments. The cell suspension directly stressed with CuCl2 was placed in compartment A and was incubated with the untreated cell suspension in compartment B. Such treatment significantly increased the levels of cell death and eATP content of the cell suspension in these two compartments. In contrast, addition of ATPase into the cell suspension directly stressed with CuCl2 decreased the eATP level in these two compartments but further increased the level of cell death in compartment B, compared to no addition of ATPase. Similar results were obtained when tobacco cell suspension was replaced by wheat seedlings. These observations indicate that when Cu stress from compartment A induced the plant cell death in compartment B, ATP transferred from compartment A could play a role in alleviating this cell death. Thus, it is suggested that eATP released by copper stress could act as diffusible signal in alleviating the Cu stress-induced cell death. [ABSTRACT FROM AUTHOR]

Hypertonic salt stress with different concentrations of NaCl increased the levels of extracellular ATP of Arabidopsis leaves. And, hypertonic salt stress decreased the levels of Fv/Fm (the maximal efficiency of photosystem II), ΦPSII (the photosystem II operating efficiency), qP (photochemical quenching), and intracellular ATP (iATP) production. The treatment with β,γ-methyleneadenosine 5′-triphosphate (AMP-PCP), which can exclude extracellular ATP from its binding sites of extracellular ATP receptors, caused a further decrease in the levels of Fv/Fm, ΦPSII,qP, and iATP production of the salt-stressed Arabidopsis leaves, while the addition of exogenous ATP rescued the inhibitory effects of AMP-PCP on ΦPSII, qP, and iATP production under hypertonic salt stress. Under hypertonic salt stress, the values of Fv/Fm, ΦPSII, qP, and iATP production were lower in the dorn 1–3 mutant than in the wild-type plants. These results indicate that the responses of photosystem II and intracellular ATP production to salt stress could be affected by extracellular ATP. [ABSTRACT FROM AUTHOR]

Although excess excitation energy (EEE) can damage the photosynthetic apparatus and deregulate many cellular processes, some studies have reported that EEE can be used by plants to optimize the resistance to pathogen infection. Here, we investigated whether the EEE-induced resistance to pathogen infection might be mediated by mitochondrial alternative oxidase (AOX). The present work showed that exposure of Arabidopsis (Arabidopsis thaliana) leaves to short-term excess light or treatment of Arabidopsis leaves with the electron transport inhibitor, DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone), restricted the growth of virulent biotrophic bacteria, Pseudomonas syringae pv tomato (Pst) DC3000. Exposure to short-term excess light or DBMIB treatment also enhanced the level of the capacity of the cyanide-resistant respiration of the infected leaves. The lack of AOX1a gene in the AtAOX1a anti-sense line attenuated the excess light- or DBMIB-induced resistance toward Pst DC3000. These results indicate that AOX may play a role in regulating the EEE-induced resistance to virulent pathogen infection. [ABSTRACT FROM AUTHOR]