Modulation of soybean and maize antioxidant activities by Caffeic acid and nitric oxide under salt stress

Philosophiae Doctor - PhDThis study explores the roles of exogenously applied nitric oxide, exogenously applied caffeic acid and salt stress on the antioxidant system in cereal (exemplified by maize) and legume (using soybean as an example) plants together with their influence on membrane integrity and cell death.This study investigates changes in H2O2 content, root lipid peroxidation, root cell death and antioxidant enzymatic activity in maize roots in response to exogenously applied nitric oxide (NO) and salt stress. This part of the study is based on the partially understood interaction between NO and reactive oxygen species (ROS) such as H2O2 and the role of antioxidant enzymes in plant salt stress responses. The results show that application of salt (NaCl) results in elevated levels of H2O2 and an increase in lipid peroxidation, consequently leading to increased cell death. The study also shows that by regulating the production and detoxification of ROS through modulation of antioxidant enzymatic activities, NO plays a pivotal role in maize responses to salt stress. The study argues for NO as a regulator of redox homeostasis that prevents excessive ROS accumulation during exposure of maize to salinity stress that would otherwise be deleterious to maize. This study extends the role of exogenously applied NO to improve salt stress tolerance in cereals crops (maize) further to its role in enhancing salt stress tolerance in legumes. The effect of long-term exposure of soybean to NO and salt stress on root nodule antioxidant activity was investigated to demonstrate the role of NO in salt stress tolerance. The results show that ROS scavenging antioxidative enzymes like SOD, GPX and GR are differentially regulated in response to exogenous application of NO and salt stress. It remains to be determined if the NOinduced changes in antioxidant enzyme activity under salt stress are sufficient to efficiently reduce ROS accumulation in soybean root nodules to levels close to those of unstressed soybean root nodules. Furthermore, this study investigates the effect of long-term exposure of soybean to exogenous caffeic acid (CA) and salt stress, on the basis of the established role of CA as an antioxidant and the involvement of antioxidant enzymes in plant salt stress responses. The effect of CA on soybean nodule number, biomass (determined on the basis of nodule dry weight, root dry weight and shoot dry weight), nodule NO content, and nodule cyclic guanosine monophosphate (cGMP) content in response to salt stress was investigated. Additionally, CA-induced changes in nodule ROS content, cell viability, lipid peroxidation and antioxidant enzyme activity as well as some genes that encode antioxidant enzymes were investigated in the presence or absence of salt stress. The study shows that long-term exposure of soybean to salt stress results in reduced biomass associated with accumulation of ROS, elevated levels of lipid peroxidation and elevated levels of cell death. However, exogenously applied CA reversed the negative effects of salt stress on soybean biomass, lipid peroxidation and cell death. CA reduced the salt stress-induced accumulation of ROS by mediating changes in root nodule antioxidant enzyme activity and gene expression. These CA-responsive antioxidant enzymes were found to be superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione peroxidase (GPX), and glutathione reductase (GR), which contributed to the scavenging of ROS in soybean nodules under salt stress. The work reported in Chapter 2 has been published in a peer-reviewed journal [Keyster M, Klein A, Ludidi N (2012) Caspase-like enzymatic activity and the ascorbate-glutathione cycle participate in salt stress tolerance of maize conferred by exogenously applied nitric oxide. Plant Signaling and Behavior 7: 349-360]. My contribution to the published paper was all the work that is presented in Chapter 2,whereas the rest of the work in the paper (which is not included in Chapter 2) was contributed by Dr Marshall Keyster

Nitric oxide (NO) regulates the expression of single-domain cystatins in glycine max (soybean)

Plant cystatins inhibit cysteine proteases and are important in regulating plant development and plant responses to biotic and abiotic stress. Furthermore, nitric oxide plays a signaling role in regulating plant responses to developmental processes, biotic and abiotic stress. With the aim of determining if nitric oxide is involved in the regulation of the expression of single-domain cystatins, we have identified single-domain cystatin genes in soybean (Glycine max cv. PAN626) on the basis of sequence homology to a nitric oxide-inducible cystatin (AtCYS1, At5g12140) from Arabidopsis thaliana. Analysis of the expression of the four cystatin genes revealed that transcript levels of these cystatins are altered by exogenously applied nitric oxide and a nitric oxide synthase inhibitor. Altered expression of these cystatins by nitric oxide and the nitric oxide synthase inhibitor implies that changes in cellular nitric oxide content, which have previously been shown to occur during development and/or biotic and abiotic stress, influence soybean physiological processes that are regulated by cysteine proteases. Recombinant protein expression of one of the cystatins (as a glutathione-S-transferase fusion protein) showed that it has inhibitory activity against the model cysteine protease papain but not the model serine protease trypsin and that it inhibits caspase-like activity in soybean nodule extracts. This serves as evidence that these four plant cystatins are functional cysteine protease inhibitors because of their high degree of primary sequence identity. It also indicates that the single-domain cystatins regulate caspase-like activity, which is known to participate in plant responses to biotic and abiotic stress. We thus conclude that nitric oxide and nitric oxide synthase-like activity regulate the expression of these cystatins, thus influencing soybean caspase-like activity. We also propose a role for this nitric oxide-mediated regulation of cystatin gene expression in the mediation of developmental processes and responses to abiotic stress in soybean.Web of Scienc

Sustainable agriculture through the enhancement of microbial biocontrol agents: Current challenges and new perspectives

The future of pesticide usage in agriculture is uncertain due to its unsustainability, adverse environmental impacts, and its association in enhanced phytopathogen resistance. Hence, this situation urges the development of new sustainable practices in agriculture. A promising approach involves endophytes, which are non-pathogenic microorganisms inhabiting the interior parts of plants. However, due to the vast diversity and complexity of plant microbiomes, a major gap has formed with regards to endophytic research and its application in phytopathogen biocontrol. The gap has mainly been increasing due to the difficulty of isolating underrepresented endophytes and due to limitation of previous genetic tools availability to further research and understand plant-microbe interaction, endophytic biocontrol capabilities and their biocontrol compounds

Enzymatic and proteomic exploration into the inhibitory activities of lemongrass and lemon essential oils against Botrytis cinerea (causative pathogen of gray mold)

Essential oils (EOs) have been demonstrated as efficacious against B. cinerea. However, the underpinning enzymatic and proteomic mechanism for these inhibitory effects is not entirely clear. Thus, this study examined the effects of lemon (Le) and lemongrass (Lg) EOs (individually and in combination) against B. cinerea based on enzymatic and proteomic analyses. Proteomics data are available via ProteomeXchange with identifier PXD038894. Both EOs (individually and in combination) displayed abilities to induce scavenging as observed with the reduction of H2O2. Measured malondialdehyde (MDA) and superoxide dismutase (SOD) activity were increased in all EOs treated B. cinerea mycelia compared to the control. Ascorbate peroxidase (APX) activity was highest in Lg treated B. cinerea (206% increase), followed by combined (Le + Lg) treatment with 73% compared to the untreated control

Venom proteomic analysis of medically important Nigerian viper Echis ocellatus and Bitis arietans snake species

Snakebite envenoming remains a neglected tropical disease which poses severe health hazard, especially for the rural inhabitants in Africa. In Nigeria, vipers are responsible for the highest number of deaths. Hydrophilic interaction liquid chromatography coupled with LC-MS/MS was used to analyze the crude venoms of Echis ocellatus (Carpet viper) and Bitis arietans (Puff adder) in order to understand their venom proteomic identities. Results obtained revealed that gel-free proteomic analysis of the crude venoms led to the identification of 85 and 79 proteins, respectively. Seventy-eight (78) proteins were common between the two snake species with a 91.8% similarity score. The identified proteins belong to 18 protein families in E. ocellatus and 14 protein families in B. arietans. Serine proteases (22.31%) and metalloproteinases (21.06%) were the dominant proteins in the venom of B. arietans; while metalloproteinases (34.84%), phospholipase A2s (21.19%) and serine proteases (15.50%) represent the major toxins in the E. ocellatus venom. Other protein families such as three-finger toxins and cysteine-rich venom proteins were detected in low proportions. This study provides an insight into the venom proteomic analysis of the two Nigerian viper species, which could be useful in identifying the toxin families to be neutralized in case of envenomation

Drought and salinity stress alters ROS accumulation, water retention, and osmolyte content in sorghum plants

biotic stresses such as drought and salinity are major environmental factors that influence crop productivity worldwide. These adverse conditions induce osmotic stresses in plant cells by decreasing water availability, thus leading to loss of cell turgor and the accumulation of reactive oxygen species (ROS) that are detrimental to plant growth and development. To survive such harsh environmental conditions, plants must initiate intracel- lular and physiological signaling networks to rapidly respond and efficiently neutralize these stresses. Inefficient scavenging of ROS would lead to increased levels of cell death, thus inhibiting plant growth and reducing crop productivity. This study investigates the effect of drought and salinity stress on plant growth, water retention, oxidative damage, chlorophyll content, and proline accumulation in sorghum plants. Plant growth, biomass, and leaf chlorophyll were significantly reduced whereas the total proline content was enhanced in response to stress conditions. The significant increase in hydrogen peroxide content as a consequence of stress conditions resulted in augmented levels of lipid peroxidation, which was manifested as extensive cell death and biomass reduction

Diagnostic approaches of pneumonia for commercial-scale biomedical applications: An overview

Pneumonia remains the leading infectious cause of death among children under five years of age, and the elderly. Several biomarkers, which have been identified for its diagnosis lack specificity, as they could not differentiate viral from bacterial pathogens of the disease; these biomarkers also fail to establish a distinction between pneumonia and other associated diseases such as pulmonary tuberculosis and Human Immunodeficiency Virus (HIV). This review outlined the menace of pneumonia disease from the statistical prevalence, clinical and immunological view, challenges with the methods used in diagnosis, and more useful information about methods of diagnosis of pneumonia with their limitations as well. Additionally, the use of aptamers and antimicrobial peptides (AMPs) rather than antibodies to bind and recognize receptors for diagnostics, offers several advantages over other biomarkers shortcomings such as non-specificity

Functional prediction of candidate micrornas for CRC management using in silico approach

Approximately 30–50% of malignant growths can be prevented by avoiding risk factors and implementing evidence-based strategies. Colorectal cancer (CRC) accounted for the second most common cancer and the third most common cause of cancer death worldwide. This cancer subtype can be reduced by early detection and patients’ management. In this study, the functional roles of the identified microRNAs were determined using an in silico pipeline. Five microRNAs identified using an in silico approach alongside their seven target genes from our previous study were used as datasets in this study. Furthermore, the secondary structure and the thermodynamic energies of the microRNAs were revealed by Mfold algorithm. The triplex binding ability of the oligonucleotide with the target promoters were analyzed by Trident. Finally, evolutionary stage-specific somatic events and co-expression analysis of the target genes in CRC were analyzed by SEECancer and GeneMANIA plugin in Cytoscape. Four of the five microRNAs have the potential to form more than one secondary structure. The ranges of the observed/expected ratio of CpG dinucleotides of these genes range from 0.60 to 1.22

Comparative venom toxin analyses of Nigerian viperidae and elapidae snakes

Envenoming by snakebite is a serious health problem that maims and kills a large number of people, primarily in rural areas of developing African countries. The first comparative venom proteomic analyses of four snakes from the viperidae (E. ocellatus and B. arietans) and elapidae (N. haje and N. katiensis) families are presented in this study. Two-dimensional electrophoresis was combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry to analyze the venoms. Proteins were identified by comparing mass spectrometry spectra to those in the reviewed Uniprot-Serpentes database. A protein spot was considered differentially present between samples at a p-value of 2. Viper venoms contained cytotoxic-inducing proteins such as SVMPs, SVSPs, and cytotoxins, whereas elapid snake venoms contained neurotoxic proteins such as PLA2, 3-FTx, and neurotoxins

Exogenous p-coumaric acid improves salvia hispanica l. Seedling shoot growth

p-Coumaric acid (p-CA) belongs to a family of natural esters of hydroxycinnamic acid compounds that have been shown to modulate plant growth and metabolism. In this study, we investigated the effect of exogenous p-CA on plant growth, reactive oxygen species (ROS)-induced oxidative damage, photosynthetic metabolism, osmolyte content and changes in superoxide dismutase (SOD) enzymatic activity