1. From a study of the system salicylic acid-biphenyl it was concluded that there was no compound formation in the system. The solution is very nearly ideal, having an eutectic temperature of 67.6℃ at a mole fraction of .903 for the biphenyl. 2. The system salicylic acid-diphenylamine was studied and no compound formation was found to be present. The system and no compound formation was found to be present. The system is not as ideal as the salicylic acid-biphenyl system, but follows the ideal solution curve fairly well. The system has an eutectic temperature of 48.5℃ at .926 mole fraction of diphenylamine. / M.S.

Abiotic stress poses a significant threat to crop productivity and food security. In this study, we focused on understanding the role of SIP-428, a SABP2-interacting protein, in mediating plant responses to environmental stresses. Transgenic tobacco plants overexpressing SIP-428 were subjected to salinity and drought stress. The overexpression of SIP-428 led to diminished growth under both stress conditions, indicating a negative impact on stress tolerance. Specifically, SIP-428 overexpression resulted in a reduction in catalase activity, while peroxidase activity remained unaffected. These findings suggest that SIP-428 plays a negative regulatory role on the catalase activity during abiotic stress, which may contribute to the susceptibility of plants to such stresses. Understanding the molecular mechanisms underlying SIP-428's role in stress responses could potentially lead to strategies for enhancing stress tolerance in crops, thereby promoting sustainable agricultural practices.

Plants synthesize a diverse array of phytochemicals in response to interactions with herbivores, pathogens, and commensal microbes. These phytochemicals may simultaneously enhance crop defense and quality, representing a potential pest management strategy. However, plant chemical responses to different types and levels of biotic interactions remain unclear, particularly in fruit tissues, and the feasibility of inducing these defenses through elicitor application in field environments also requires further examination. Thus, apples were used to 1) examine the impact of distinct communities of biotic interactions among plants, insects, and microbes on fruit phenolic chemistry, and 2) examine the impact of the phytohormones jasmonic acid (JA), salicylic acid (SA), and melatonin (M) on fruit phenolic chemistry and resistance against pests and pathogens. Ultimately, phenolic defenses were induced by fungal damage primarily in ripe pulp tissues, where there was also a positive relationship between fungal endophyte and phenolic diversity, supporting a broad hypothesis that chemical diversity may increase with biotic diversity. Additionally, two compounds were upregulated in response to fungal damage: chlorogenic acid and an unidentified benzoic acid. Elicitor applications did not affect phenolic chemistry, but the combined application of JA-SA analogues had some chemical or physical effect, as this treatment reduced emergence of the insect Rhagoletis pomonella. Thus, fruit induced defenses may be tissue-specific and subject to temporal, environmental, or genotypic variation. Overall, these chapters examined the relationship between biotic interactions and induced fruit chemistry, with the goal of improving understanding of plant-microbe-insect interactions and incorporating these interactions into more sustainable agricultural practices. / Master of Science / Plants may produce a diverse array of defensive phytochemical compounds in response to interactions with herbivores, pathogens, and the microorganisms that reside within plant tissues. These phytochemicals may simultaneously improve crop defenses and quality, representing a potential agricultural management strategy. However, plant chemical responses to different types and levels of biotic interactions are not well-understood, particularly in fruit tissues, and the feasibility of activating these defenses in fruits through the application of phytohormones that regulate defense pathways as a potential management strategy also requires further examination. Thus, apples were used to 1) examine the impact of distinct communities of biotic interactions among plants, insects, and microbes on fruit chemistry, focusing on phenolics, an important class of phytochemical compounds, and 2) examine the impact of the defense-activating phytohormones jasmonic acid (JA), salicylic acid (SA), and melatonin (M) on fruit phenolic chemistry and resistance against pests and pathogens. Ultimately, phenolic defenses were activated by fungal damage primarily in ripe pulp tissues, where there was also a positive relationship between fungal endophyte and phenolic diversity, supporting a broad hypothesis that chemical diversity may increase with biotic diversity. Additionally, two compounds were produced in response to fungal damage: chlorogenic acid and an unidentified benzoic acid. Phytohormone applications did not affect phenolic chemistry, but the application of the combined JA-SA analogues had some chemical or physical effect, as this treatment reduced emergence of the insect Rhagoletis pomonella. Overall, the phytochemical defenses activated by biotic interactions in fruits may occur primarily in certain tissue types, and may also vary due to environmental conditions, time of year, or plant species. These chapters examined the relationship between fruit chemistry and biotic interactions with the goal of improving understanding of plant-microbe-insect interactions and incorporating these interactions into more sustainable agricultural practices.

Salicylic acid binding protein 2 (SABP2) is known to play a key role in Salicylic acid mediated defense pathway. SBIP-470 is SABP2 interacting protein that might be putatively involved in transfer of lipids. SBIP-470 was cloned without the signal peptide and expressed in E. coli. In vitro lipid binding assay using recombinant SBIP-470 failed to detect lipid binding. In vitro lipid transfer assay showed recombinant SBIP-470 does not transfer phospholipid. Study has shown that SBIP-470 is highly inducible upon infection with viral as well as bacterial pathogens. Induction of SBIP-470 expression upon the TMV infection most likely depends upon the SABP2 while its expression upon non-host bacterial pathogens is most probably inhibited by the SABP2. A study of Arabidopsis knockout mutants (ltp12 mutant and ltp2 mutant) lacking the SBIP-470 homolog genes showed defects in growth phenotype, and they were found susceptible to bacterial pathogens.

Tobacco SABP2, a 29. kDa protein catalyzes the conversion of methyl salicylic acid (MeSA) into salicylic acid (SA) to induce SAR. Pretreatment of plants with acibenzolar-. S-methyl (ASM), a functional analog of salicylic acid induces systemic acquired resistance (SAR). Data presented in this paper suggest that SABP2 catalyzes the conversion of ASM into acibenzolar to induce SAR. Transgenic SABP2-silenced tobacco plants when treated with ASM, fail to express PR-1 proteins and do not induce robust SAR expression. When treated with acibenzolar, full SAR is induced in SABP2-silenced plants. These results show that functional SABP2 is required for ASM-mediated induction of resistance.

Tobacco SABP2 has been shown to display high affinity for salicylic acid (SA) and methylsalicylate (MeSA) and plays an important role in SAR signal development. Using biochemical approach, SABP2 has been shown to demonstrate strong esterase activity in converting MeSA to SA. Recent study shows that tetra fluoroacetophenone, a synthetic analog of SA, competitively inhibits SABP2 esterase activity as well as suppresses SAR signal development in tobacco mosaic virus (TMV)-infected tobacco plants. Not much has been studied on the effect of pesticides on plant defenses. Because both AChE and SABP2 are esterase-like proteins belonging to α/β hydroxylase superfamily, we hypothesize that pesticides may inhibit the MeSA esterase activity of SABP2 and block SAR development. Biochemical and molecular biology techniques were used to test this hypothesis. SAR in tobacco-TMV plant-pathogen system is measured by significant decrease in TMV-induced lesion sizes in secondarily inoculated distal leaves.

Thesis (M.Sc. (Horticulture)) -- University of Limpopo, 2022 / The South African Avocado Industry has recently announced plans to expand exports into new markets, such as the United States (US). As a requirement for these markets, fruit of high quality must be stored at ultra-low temperature to mitigate phytosanitary risks. However, ‘Hass’ avocado fruit are susceptible to chilling injury when stored at temperatures below 3°C. Moreover, CI development resulted in uneven ripening and disease infestation due to damaged cell membranes. Therefore, the objective of this study was to evaluate the potential of methyl jasmonate (MeJA) and salicylic acid (SA) on quality maintenance of 'Hass' avocado fruit during ultra-low cold storage. Matured ‘Hass’ avocado fruit were harvested at commercial dry matter (22%). The experiment was conducted using a completely randomized design (CRD) with eight replications per treatment. Treatment concentrations for methyl jasmonate (MeJA) were 0 (control), 10 and 100 μmol•L−1 , while those for salicylic acid (SA) were 0 (control), 1.0, 2.0 and 3.0 mM. After treatments, fruit were stored at 2°C for 31 days and thereafter, ripened at ambient temperature (±25°C) until fully ripe. During ripening, fruit were evaluated for weight loss, exocarp colour, firmness, chilling injury, as well as physiological (vascular browning) and pathological disorders (fruit rot). In this study, dipping fruit in MeJA solution significantly (P < 0.05) reduced ‘Hass’ avocado fruit firmness loss. Moreover, MeJA showed a significant effect (P < 0.05) on hue angle (h°) but did not significantly affect (P > 0.05) visual colour rating, chroma (C*), lightness (L*) and weight loss. The results showed that ‘Hass’ avocado fruit treated with 10 μmol•L−1 MeJA reduced weight loss when compared with 100 μmol•L−1 MeJA from day 2 to day 8 of ripening. Overall results showed a visual change in ‘Hass’ avocado fruit exocarp colour, with eye colour changing from rating 1 (emerald-green) to 3 (olive- green) for control and fruit treated with MeJA throughout the ripening days. Furthermore, MeJA reduced ‘Hass’ avocado fruit external chilling injury, physiological and pathological disorders. With respect to SA treatments, the result showed that dipping fruit at 1.0 and 2.0 mM SA had a significant effect (P < 0.05) on reducing firmness loss during ripening. Salicylic acid (1.0 mM) reduced and alleviated ‘Hass’ avocado fruit external chilling injury during ultra-low cold storage. Furthermore, result showed that 1.0 and 2.0 mM SA treatments had significant affect (P < 0.05) on firmness loss. Moreover, a significant effect was observed on visual colour and C* but did not affect (P > 0.05) L* and h°. Fruit treated with SA showed poor exocarp colour development with extended exposure to ultra-low cold storage, as a result, developed chilling symptoms. The treatment of ‘Hass’ avocado fruit with 1.0 mM SA inhibited the incidence of fruit rot and vascular browning when compared with control and fruit treated with 2.0 and 3.0 mM SA. In conclusion, 10 and 100 μmol•L−1 MeJA and 1.0, 2.0 and 3.0 mM SA effectively preserved ‘Hass’ avocado fruit quality during storage at ultra-low temperature. / Agricultural Sector Education Training Authority (AgriSeta) and National Research Foundation (NRF)

Salicylic Acid Binding Protein 2 (SABP2) is a 29kDa protein present in extremely low amounts in tobacco leaves. SABP2 processes the mobile defense signal, methyl salicylic acid generated in plants resisting microbial infection. The precise localization of SABP2 in plants is not known. SABP2 has not been shown to have any targeting signal peptides. This study was designed to determine localization of SABP2 in tobacco plants. Biochemical and immunological studies using antibodies against SABP2 suggest that it is localized to the chloroplast, associating with chloroplast envelope membranes. Chloroplast import assays confirm that SABP2 is associated with the chloroplast envelope membrane. Solubilization and analysis of chloroplast membrane proteins show that imported SABP2 associates with the chloroplast envelope membrane by weak hydrophobic and/or ionic interactions. Cellular localization and understanding mechanisms of SABP2 import to the chloroplast will be important from a metabolic engineering standpoint to enhance plant natural defense against microbial pathogens.

Salicylic acid binding protein 2 (SABP2) is one of the key enzymes in salicylic acid-dependent plant defense pathway. SABP2 is a 29 kDa protein present in extremely low abundance in plants and it catalyzes the conversion of signaling molecule methyl salicylate into salicylic acid. Although it has been shown that 6x His-tagged SABP2 over expressed in E. coli is a homodimer, its exact conformation in planta is still unknown. Therefore, we proposed to determine if SABP2 exist as a dimer and/or monomer under natural condition. To verify the exact conformation of native SABP2 protein in plant, SABP2 was purified from wild type tobacco using a 5-step purification protocol. Analysis of purified SABP2 in gel filtration and immunoblot assay suggested that SABP2 exists as a monomer in tobacco plant. Studies on SABP2 conformation will give us insight into the structure and functional relationship of this protein in salicylic acid-dependent disease resistance pathway.

SABP2 (Salicylic Acid Binding Protein 2) plays a vital role in the salicylic acid signaling pathway of plants both regarding basal resistance and systemic acquired resistance against pathogen infection. SIP-428 (SABP2 Interacting Protein-428) is a Silent information regulator 2 (SIR2) like deacetylase enzyme that physically interacts with SABP2 in a yeast two-hybrid interaction and confirmed independently by a GST pull-down assay. We demonstrated that SIP- 428 is an NAD+ dependent SIR2 deacetylase enzyme. Transgenic tobacco plants silenced in SIP- 428 expression via RNAi showed enhanced basal resistance to microbial pathogens. Moreover, these SIP-428-silenced lines also exhibited a robust induction of systemic acquired resistance. In contrast, the transgenic tobacco lines overexpressing SIP-428 showed compromised basal resistance and failed to induce systemic acquired resistance. These results indicate that SIP-428 is likely a negative regulator of SA-mediated plant immunity. Experiments using a SABP2 inhibitor showed that SIP-428 likely functions upstream of SABP2 in the salicylic acid signaling pathway. It also indicates that SABP2 is dependent on SIP-428 for its role in the SA signaling pathway. Subcellular localization studies using confocal microscopy and subcellular fractionation showed that SIP-428 localized in the mitochondria. These results clearly show a role for SIP-428 in plant immunity.