Marshall Keyster

Nitric oxide increases the enzymatic activity of three ascorbate peroxidase isoforms in soybean root nodules

Ascorbate peroxidase is one of the major enzymes regulating the levels of H2O2 in plants and plays a crucial role in maintaining root nodule redox status. We used fully developed and mature nitrogen fixing root nodules from soybean plants to analyse the effect of exogenously applied nitric oxide, generated from the nitric oxide donor 2,2′-(hydroxynitrosohydrazono)bis-ethanimine, on the enzymatic activity of soybean root nodule ascorbate peroxidase. Nitric oxide caused an increase in the total enzymatic activity of ascorbate peroxidase. The nitric oxide-induced changes in ascorbate peroxidase enzymatic activity were coupled to altered nodule H2O2 content. Further analysis of ascorbate peroxidase enzymatic activity identified three ascorbate peroxidase isoforms for which augmented enzymatic activity occurred in response to nitric oxide. Our results demonstrate that nitric oxide regulates soybean root nodule ascorbate peroxidase activity. We propose a role of nitric oxide in regulating ascorbate-dependent redox status in soybean root nodule tissue.

Caspase-like enzymatic activity and the ascorbate-glutathione cycle participate in salt stress tolerance of maize conferred by exogenously applied nitric oxide

Salinity stress causes ionic stress (mainly from high Na+ and Cl- levels) and osmotic stress (as a result of inhibition of water uptake by roots and amplified water loss from plant tissue), resulting in cell death and inhibition of growth and ultimately adversely reducing crop productivity. In this report, changes in root nitric oxide content, shoot and root biomass, root H2O2 content, root lipid peroxidation, root cell death, root caspase-like enzymatic activity, root antioxidant enzymatic activity and root ascorbate and glutathione contents/redox states were investigated in maize (Zea mays L. cv Silverking) after long-term (21 d) salt stress (150 mM NaCl) with or without exogenously applied nitric oxide generated from the nitric oxide donor 2,2′-(Hydroxynitrosohydrazano)bis-ethane. In addition to reduced shoot and root biomass, salt stress increased the nitric oxide and H2O2 contents in the maize roots and resulted in elevated lipid peroxidation, caspase-like activity and cell death in the roots. Altered antioxidant enzymatic activities, along with changes in ascorbate and glutathione contents/redox status were observed in the roots in response to salt stress. The detrimental effects of salt stress in the roots were reversed by exogenously applied nitric oxide. These results demonstrate that exogenously applied nitric oxide confers salt stress tolerance in maize by reducing salt stress-induced oxidative stress and caspase-like activity through a process that limits accumulation of reactive oxygen species via enhanced antioxidant enzymatic activity.

Endogenous NO levels regulate nodule functioning Potential role of cGMP in nodule functioning

Nitric oxide is a small gaseous signaling molecule which functions in the regulation of plant development and responses to biotic and abiotic stresses. Recently, we have shown that nitric oxide is required for development of functional nodules. Here, we show that inhibition of nitric oxide synthase enzymatic activity (using Nω-nitro-L-arginine) reduces nitric oxide content in soybean root nodules and this is coupled by reduction of endogenous cyclic guanosine monophosphate content in the nodules. We postulate that the regulation of soybean nodule development by nitric oxide is transduced via cyclic guanosine monophosphate through activation of nitric oxide-responsive soluble guanylate cyclase. Furthermore, we hypothesize that this signaling cascade is mediated via modulation of the activities of antioxidant metabolic pathways.

Nitric oxide affects salt-induced changes in free amino acid levels in maize

It was assumed that salt-induced redox changes affect amino acid metabolism in maize (Zea mays L.), and this influence may be modified by NO. The applied NaCl treatment reduced the fresh weight of shoots and roots. This decrease was smaller after the combined application of NaCl and an NO-donor ((Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate, DETA/NO) in the shoots, while it was greater after simultaneous treatment with NaCl and nitro-L-arginine (L-NNA, inhibitor of NO synthesis) in the roots. The quantum yield efficiency of photosystem II was not influenced by the treatments. NaCl had a significant effect on the redox environment in the leaves as it was shown by the increase in the amount of glutathione disulphide and in the redox potential of the glutathione/glutathione disulphide redox pair. This influence of NaCl was modified by DETA/NO and L-NNA. Pharmacological modification of NO levels affected salt-induced changes in both the total free amino acid content and in the free amino acid composition. NaCl alone increased the concentration of almost all amino acids which effect was strengthened by DETA/NO in the case of Pro. L-NNA treatment resulted in a significant increase in the Ala, Val, Gly and Tyr contents. The Ile, Lys and Val concentrations rose considerably after the combined application of NaCl and DETA/NO compared to NaCl treatment alone in the recovery phase. NaCl also increased the expression of several genes related to the amino acid and antioxidant metabolism, and this effect was modified by DETA/NO. In conclusion, modification of NO levels affected salt-induced, glutathione-dependent redox changes and simultaneously the free amino acid composition and the level of several free amino acids. The observed much higher Pro content in plants treated with both NaCl and DETA/NO during recovery may contribute to the protective effect of NO against salt stress.

Inhibition of NOS- like activity in maize alters the expression of genes involved in H2O2 scavenging and glycine betaine biosynthesis

Nitric oxide synthase-like activity contributes to the production of nitric oxide in plants, which controls plant responses to stress. This study investigates if changes in ascorbate peroxidase enzymatic activity and glycine betaine content in response to inhibition of nitric oxide synthase-like activity are associated with transcriptional regulation by analyzing transcript levels of genes (betaine aldehyde dehydrogenase) involved in glycine betaine biosynthesis and those encoding antioxidant enzymes (ascorbate peroxidase and catalase) in leaves of maize seedlings treated with an inhibitor of nitric oxide synthase-like activity. In seedlings treated with a nitric oxide synthase inhibitor, transcript levels of betaine aldehyde dehydrogenase were decreased. In plants treated with the nitric oxide synthase inhibitor, the transcript levels of ascorbate peroxidase-encoding genes were down-regulated. We thus conclude that inhibition of nitric oxide synthase-like activity suppresses the expression of ascorbate peroxidase and betaine aldehyde dehydrogenase genes in maize leaves. Furthermore, catalase activity was suppressed in leaves of plants treated with nitric oxide synthase inhibitor; and this corresponded with the suppression of the expression of catalase genes. We further conclude that inhibition of nitric oxide synthase-like activity, which suppresses ascorbate peroxidase and catalase enzymatic activities, results in increased H2O2 content.

Sustainable Nanotechnology: Mycotoxin Detection and Protection

Advances in nanotechnology have demonstrated vast applications due to the properties of nanomaterials. One of the applications that have emerged is the utilization of nanotechnology in agriculture with the emphasis on precision agricultural practices. That is, using nanoformulations to maximize crop production while minimizing the use of pesticides and herbicides. The high surface-to-volume ratio of nanoparticles provides an improved matrix for the immobilization of desired biomolecules for signal amplification in biosensors for the detection of mycotoxins. Metallic and magnetic nanoparticles are widely used in the fabrication of nanosensors for mycotoxin detection. These nanoparticles are used in nanocomposite material for the production of nanopackaging to increase shelf life of agricultural produce. Green nanotechnology formulations of using phytochemicals from plant material to produce nanofungicides are feasible in the agriculture due to no toxic effect towards human and animal health. The present chapter provides recent work carried out in the field of nanotechnology for detection of mycotoxins and highlights some of the commercial nanoformulations used in agriculture.