Browsing by Author "Adams, Ruqaiyah"

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    Characterization of a novel soybean candidate glutathione peroxidase/thioredoxin-dependent peroxidase under salt stress
    (2012) Adams, Ruqaiyah; Ludidi, N. N.; Keyster, Marshall
    The production of reactive oxygen species (ROS) is prominent in all aerobic metabolisms including plants. For this reason, the redox homeostasis of the production and scavenging of these intermediates is imperative for growth, development and survival during unfavourable conditions. In this study, a putative glutathione peroxidase gene (Glyma17g34110) from Glycine max (soybean) was identified and analyzed. The successful characterisation of Glyma17g34110 provided evidence of it being a glutathione peroxidase using glutathione as its preferred electron donor and substrate. Furthermore, it is known that antioxidant enzymes such as GPX exist in various tissues, performing a diverse set of functions. By a bioinformatic analysis of Glyma17g34110 and its promoter region, it was indicated that Glyma17g34110 could be a putative chloroplast protein that could play an important role in photosynthesis.One of the major factors affecting plant growth and development worldwide is abiotic stresses such as salinity. In the presence of salinity the production of harmful ROS is increased, resulting in detrimental reactions with important biological features (DNA, protein and lipid membranes), leading to cell death. The analysis of Glyma17g34110 under salt stress revealed that it is a salt sensitive gene and thus, the down-regulation of Glyma17g34110 could be due to the lack of known defence and response cis-acting elements present in the promoter region. Furthermore, it was proven in previous studies that the application of exogenous nitric oxide (NO) increases the activity of antioxidant enzymes. In this thesis it was observed that the presence of exogenously applied NO increased the expression of Glyma17g34110 tremendously in all soybean tissues (leaves, roots and nodules) investigated.Studies have found numerous cis-acting elements to be NO responsive, however, none of these elements were found in the promoter region upstream of glyma17g34110. This suggests that novel cis-acting elements could be present in the promoter region of Glyma17g34110.Thus, increasing the expression of Glyma17g34110 during salinity in the presence of NO, as well as the identification of these novel cis-acting elements, could lead to the enhancement of the defence mechanisms against ROS, which could lead to increasing plant tolerance to stress.
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    Nitric oxide (NO) regulates the expression of single-domain cystatins in glycine max (soybean)
    (Southern Cross Publishing, 2013) Keyster, Marshall; Adams, Ruqaiyah; Klein, Ashwil; Ludidi, Ndiko
    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.