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Item Open Access Exploring edamame survival mechanisms under combined drought and heat stress: photosynthesis efficiency and carbohydrate accumulation(Elsevier, 2024) Hlahla, Jeremiah M.; Mafa, Mpho S.; van der Merwe, Rouxléne; Moloi, Makoena J.The combined drought and heat (DH) stress have devastating effects on plant physiological and biochemical processes, leading to poor yield. The aims of this study were to identify the physio-biochemical mechanisms employed by edamame (Glycine max L. Merrill) for survival and to establish better performing cultivars under DH stress. The impact of DH stress on the photosynthesis efficiency and osmolytes production in three edamame cultivars (UVE14, UVE17 and AGS429) was investigated. Non-destructive measurements were performed to determine the photosynthesis attributes, while pigments, non-structural carbohydrates (starch, glucose, sucrose, trehalose) and proline were extracted and quantified spectrophotometrically. The results showed that chlorophyll a (Chl-a), Chl-b, total chlorophyll and carotenoids in AGS429 were not affected under DH stress, which corresponded to increased normalised difference vegetative index (NDVI). Positive correlations between the carotenoids and total chlorophyll contents suggest that in AGS429, prevention of chlorophyll degradation under DH stress could be attributed to the increased carotenoids because they have antioxidative function. Additionally, AGS429 and UVE14 had increased trehalose, suggesting high osmotic adjustment under DH stress. An increase in starch production maintained glucose balance in AGS429 and UVE14, demonstrating higher photosynthetic efficiency compared to UVE17. The DH stress reduced photochemical reactions and carbohydrate accumulation in UVE17. This study shows that AGS429 and UVE14 protected the photosystems and photosynthetic pigments during DH stress, which led to higher photosynthetic capacity and accumulation of carbohydrates. Thus, maintaining the photosynthesis efficiency and carbohydrate metabolism processes in the AGS429 and UVE14 were adaptation features under DH stress.Item Open Access Sustainable agriculture through the Enhancement of Microbial Biocontrol Agents: Current Challenges and New Perspectives(MDPI, 2023) Gokul, Arun; Mabaso, Jabulani; Henema, Nontuthuko; Otomo, Laetitia; Bakare, Olalekan O.; Klein, Ashwil; Daniel, Augustine I.; Omolola, Aina; Niekerk, Lee-Ann; Nkomo, Mbukeni; Keyster, MarshallThe 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. This review highlights the current challenges being encountered in this research field. Additionally, the research advances through utilization of specialized techniques (CRISPR/Cas9 system, nanoparticles and multi-omics) are highlighted to assist in elucidating the mechanism revolving around plant-microbe interactions and to generate model systems demonstrating improved biocontrol capabilities of endophytes. The ultimate goal of this review is to provide improved approaches that could be implement in an array of microorganism that will enhance the phytopathogen biocontrol field in order to create a sustainable agricultural sector.Item Open Access Regulation of proline accumulation and protein secretion in sorghum under combined osmotic and heat stress(MDPI, 2024) Ngwenya, Samkelisiwe P.; Moloi, Sellwane J.; Shargie, Nemera G.; Brown, Adrian P.; Chivasa, Stephen; Ngara, RudoPlants reprogramme their proteome to alter cellular metabolism for effective stress adaptation. Intracellular proteomic responses have been extensively studied, and the extracellular matrix stands as a key hub where peptide signals are generated/processed to trigger critical adaptive signal transduction cascades inaugurated at the cell surface. Therefore, it is important to study the plant extracellular proteome to understand its role in plant development and stress response. This study examined changes in the soluble extracellular sub-proteome of sorghum cell cultures exposed to a combination of sorbitol-induced osmotic stress and heat at 40 °C. The combined stress significantly reduced metabolic activity and altered protein secretion. While cells treated with osmotic stress alone had elevated proline content, the osmoprotectant in the combined treatment remained unchanged, confirming that sorghum cells exposed to combined stress utilise adaptive processes distinct from those invoked by the single stresses applied separately. Reactive oxygen species (ROS)-metabolising proteins and proteases dominated differentially expressed proteins identified in cells subjected to combined stress. ROS-generating peroxidases were suppressed, while ROS-degrading proteins were upregulated for protection from oxidative damage. Overall, our study provides protein candidates that could be used to develop crops better suited for an increasingly hot and dry climate.Item Open Access The roles of plant proteases and protease inhibitors in drought response: a review(Frontiers, 2023) Moloi, Sellwane Jeanette; Ngara, RudoUpon exposure to drought, plants undergo complex signal transduction events with concomitant changes in the expression of genes, proteins and metabolites. For example, proteomics studies continue to identify multitudes of drought-responsive proteins with diverse roles in drought adaptation. Among these are protein degradation processes that activate enzymes and signalling peptides, recycle nitrogen sources, and maintain protein turnover and homeostasis under stressful environments. Here, we review the differential expression and functional activities of plant protease and protease inhibitor proteins under drought stress, mainly focusing on comparative studies involving genotypes of contrasting drought phenotypes. We further explore studies of transgenic plants either overexpressing or repressing proteases or their inhibitors under drought conditions and discuss the potential roles of these transgenes in drought response. Overall, the review highlights the integral role of protein degradation during plant survival under water deficits, irrespective of the genotypes’ level of drought resilience. However, drought-sensitive genotypes exhibit higher proteolytic activities, while drought-tolerant genotypes tend to protect proteins from degradation by expressing more protease inhibitors. In addition, transgenic plant biology studies implicate proteases and protease inhibitors in various other physiological functions under drought stress. These include the regulation of stomatal closure, maintenance of relative water content, phytohormonal signalling systems including abscisic acid (ABA) signalling, and the induction of ABA-related stress genes, all of which are essential for maintaining cellular homeostasis under water deficits. Therefore, more validation studies are required to explore the various functions of proteases and their inhibitors under water limitation and their contributions towards drought adaptation.Item Open Access Green synthesis of Iron oxide and Iron dioxide nanoparticles using Euphorbia tirucalli: characterization and antiproliferative evaluation against three breast cancer cell lines(Taylor and Francis Group, 2023) Kgosiemang, Ipeleng Kopano Rosinah; Adegoke, Ayodeji Mathias; Mashele, Samson Sitheni; Sekhoacha, Mamello PatienceResearchers have become increasingly interested in nanoparticles made from plants because of their stability and large surface area. In the current study, iron oxide and iron dioxide nanoparticles were synthesized using aerial parts of the E. tirucalli as a reducing agent. The nanoparticles were analyzed using various techniques, including Ultraviolet-visible spectroscopy, Fourier Transform Infrared spectroscopy, X-ray diffractometer, X-ray photoelectron spectroscopy, X-ray energy dispersive spectroscopy, Scanning electron Microscopy, and Transmission Electron Microscopy. The nanoparticles were then investigated for their antiproliferative effect against MCF-7, SK-BR-3, MDA-MB231, and Vero cell lines. The results confirmed the formation of FeO and FeO2 nanoparticles by color change and a UV absorbance peak between 220–390 nm. EDS analysis showed traces of Fe and O, while TEM confirmed the nanoparticle size of 100 nm. FTIR showed a peak at 514 nm. The FeO-RT NPs demonstrated over 80% antiproliferative activity against the MCF-7 cell line at a concentration of 10 μg/mL. while doxorubicin, FeO-RT NPs, and DCM extract showed similar activity against the MDA-MB231 cell line at 10 and 1 g/mL concentrations. However, Vero and SK-BR-3 cell lines showed decreased antiproliferative activity. This study highlights the environmentally friendly use and safe application of iron oxide NPs in cancer therapy.Item Open Access Heterosis and combining ability of iron, zinc and their bioavailability in maize inbred lines under low nitrogen and optimal environments(Cell Press, 2023) Akhtar, Sajjad; Mekonnen, Tesfaye Walle; Mashingaidze, Kingstone; Osthoff, Gernot; Labuschagne, MarykeIron (Fe) and zinc (Zn) nutrient enrichment of staple crops through biofortification can contribute to alleviating micronutrient deficiency in sub-Saharan Africa. A line × tester mating design was used to determine the general combining ability (GCA), specific combining ability (SCA) and heterosis for grain yield, iron, Zn and phytic concentration of six lines crossed with three testers. Lines and testers were selected for high, intermediate and low mineral content. The F1 hybrids and parental lines were evaluated under low nitrogen (N) and optimum conditions across four environments over two seasons. Under low N conditions, Fe and Zn concentration in grain, and grain yield of genotypes were reduced by 9%, 9%, and 59%, respectively. However, phytic acid concentration in grain was increased by 10% under low N conditions. Both additive and non-additive gene effects were important in controlling Fe, Zn and phytic acid concentration in grain and grain yield of maize under both N conditions. The preponderance of GCA effects indicates the importance of additive gene effects in the inheritance of grain yield. Line GCA effects were more sensitive to N conditions across the environments than the tester GCA. High and significant positive SCA effects for grain yield, Fe and Zn content under low N conditions, would be a good indicator of possible heterosis in these traits. Hybrid CBY101 LM-1600 × CBY358 LM-1857 had high and significant positive SCA for grain yield under low N conditions and is a promising candidate for production in low N environments. CBY358 LM-1857 (tester) and CBY102 LM-1601 (line) are a good general combiners for Fe, Zn and GY can be used as parents in future maize hybrid breeding programs to develop high-yielding maize genotypes with high Fe and Zn content.Item Open Access Combining ability estimates for quality and non-quality protein maize inbred lines for grain yield, agronomic, and quality traits(Frontiers, 2023) Amegbor, Isaac Kodzo; Van Biljon, Angeline; Shargie, Nemera Geleta; Tarekegne, Amsal; Labuschagne, Maryke T.Introduction: Biofortified maize varieties could contribute to the fight against hunger and malnutrition of the increasing human population and help meet the high demand of maize for human consumption, industrial use and feed for animal and poultry. The understanding of the genetic mechanisms conditioning the inheritance of grain yield and other agronomic and quality traits is essential in the development of superior maize genotypes. The main objective of this study was to determine the combining ability for grain yield and other agronomic traits of QPM and non-QPM inbred lines crossed with two QPM and two non-QPM testers. Materials and methods: A total of 130 hybrids were obtained by crossing 10 non-QPM and 23 QPM lines with four elite testers (two QPM and two non-QPM). The 130 single cross hybrids were evaluated at 13 sites in 2018 and 2019. Results and discussion: The results showed significant general combining ability (GCA) for lines and testers as well as significant specific combining ability (SCA) for hybrids for most of the measured traits. Non-additive gene action controlled the inheritance of grain yield while agronomic and quality traits were controlled by additive gene action. QPM lines 11, 14 and 28 and non-QPM tester CML444 showed desirable GCA effects for grain yield, indicating that these lines in combination with tester CML444 should be considered when targeting development of superior maize genotypes with QPM traits. Conclusions: Based on the SCA values, crosses 120, 108, 105, 99, 85, and 41 were identified as the best hybrids across the locations. It is recommended that maize breeding programs targeting the development of high yielding QPM hybrids should exploit QPM lines with high GCA values for heterosis to be realized. In addition, the identified superior hybrids may be further evaluated and consequently promoted for commercial release which could increase incomes of farmers and help to alleviate poverty, hunger and malnutrition in southern Africa and sub-Saharan Africa at large.Item Open Access The effect of selenium foliar application on the physiological responses of edamame under different water treatments(MDPI, 2022) Moloi, Makoena Joyce; Khoza, Bongiwe MinahDrought has devastating effects on crops, posing enormous risks to food security. This study investigated the impact of foliar applied selenium [at four concentrations (25, 50 and 75 mg/L)] on the photosynthesis capacity, antioxidative enzyme activities [ascorbate peroxidase (APX) and guaiacol peroxidase (GPX)] and yield parameters of a drought susceptible edamame under optimal watering and drought-stressed conditions. The study was conducted in the greenhouse under controlled conditions with leaf sampling done at vegetative, flowering and pod filling stages. Treatment of drought-stressed plants with selenium selectively induced PIabs and chlorophyll content at the vegetative stage. Ascorbate peroxidase was the only parameter induced at the flowering stage by selenium under drought stress. Selenium had no effect on all parameters under drought stress at pod filling, suggesting that the efficacy of selenium declines with time. In addition, yield parameters were not substantially affected by selenium under drought stress. Although selenium was effective for selected parameters, the application should only be limited to edamame growing under drought stress because, under well-watered conditions, it had negative impacts. Future studies should explore the responses drought stressed edamame after secondary application of selenium (i.e., at vegetative, flowering and pod filling).Item Open Access Breeding of vegetable cowpea for nutrition and climate resilience in Sub-Saharan Africa: progress, opportunities, and challenges(MDPI, 2022) Mekonnen, Tesfaye Walle; Gerrano, Abe Shegro; Mbuma, Ntombokulunga WedyCurrently, the world population is increasing, and humanity is facing food and nutritional scarcity. Climate change and variability are a major threat to global food and nutritional security, reducing crop productivity in the tropical and subtropical regions of the globe. Cowpea has the potential to make a significant contribution to global food and nutritional security. In addition, it can be part of a sustainable food system, being a genetic resource for future crop improvement, contributing to resilience and improving agricultural sustainability under climate change conditions. In malnutrition prone regions of sub-Saharan Africa (SSA) countries, cowpea has become a strategic dryland legume crop for addressing food insecurity and malnutrition. Therefore, this review aims to assess the contribution of cowpea to SSA countries as a climate-resilient crop and the existing production challenges and perspectives. Cowpea leaves and immature pods are rich in diverse nutrients, with high levels of protein, vitamins, macro and micronutrients, minerals, fiber, and carbohydrates compared to its grain. In addition, cowpea is truly a multifunctional crop for maintaining good health and for reducing non-communicable human diseases. However, as a leafy vegetable, cowpea has not been researched and promoted sufficiently because it has not been promoted as a food security crop due to its low yield potential, susceptibility to biotic and abiotic stresses, quality assurance issues, policy regulation, and cultural beliefs (it is considered a livestock feed). The development of superior cowpea as a leafy vegetable can be approached in different ways, such as conventional breeding and gene stacking, speed breeding, mutation breeding, space breeding, demand-led breeding, a pan-omics approach, and local government policies. The successful breeding of cowpea genotypes that are high-yielding with a good nutritional value as well as having resistance to biotics and tolerant to abiotic stress could also be used to address food security and malnutrition-related challenges in sub-Saharan Africa.Item Open Access Expression of nutritional traits in vegetable cowpea grown under various South African agro-ecological conditions(MDPI, 2022) Gerrano, Abe Shegro; Mbuma, Ntombokulunga W.; Mumm, Rita H.Cowpea (Vigna unguiculata L.), a traditional legume food crop indigenous to Africa, has potential as both a vegetable and grain crop in contributing to dietary diversity to support health and address malnutrition, especially for those relying heavily on wheat, maize, and rice. The expression of nutritional traits (protein content and concentrations of iron (Fe), zinc (Zn), and manganese (Mn)) in cowpea leaves was evaluated over diverse agro-ecologies of South Africa and typical agronomic practices of smallholder farmers. The genotypes evaluated displayed genetic variation for all four traits. The mean values of Fe, Zn, Mn and protein content varied from 33.11 to 69.03 mg.100.g−1; 4.00 to 4.70 mg.100.g−1; and 14.40 to 19.63 mg.100.g−1 and 27.98 to 31.98%, respectively. The correlation analysis revealed significant degree of positive association between protein and Zn (r = 0.20), while negative associations were observed between Mn and protein (−0.46) and between Mn and Fe (r = −0.27). Furthermore, the expression of these important nutrient traits was influenced by the climatic conditions represented by six environments (location by year combinations) as is typical of ‘quality’ traits. Additionally, genotype-by-environment interaction effects were detected, suggesting that local soil properties and soil health may play a role in nutritional content in plants, perhaps particularly for legume crops that rely on symbiotic relationships with soil bacterial populations to fix nitrogen, which is crucial to protein formation. Further studies are needed to understand how to coordinate and align agronomic and soil management practices in vegetable cowpea production, especially those workable for the smallholder farmer, to realize the full genetic potential and nutritional value of improved vegetable cowpea varieties.Item Open Access Unraveling synergism between various GH family xylanases and deb ranching enzymes during hetero-xylan degradation(MDPI, 2021) Malgas, Samkelo; Mafa, Mpho S.; Mathibe, Brian N.; Pletscke, Brett I.Enzymes classified with the same Enzyme Commission (EC) that are allotted in different glycoside hydrolase (GH) families can display different mechanisms of action and substrate specificities. Therefore, the combination of different enzyme classes may not yield synergism during biomass hydrolysis, as the GH family allocation of the enzymes influences their behavior. As a result, it is important to understand which GH family combinations are compatible to gain knowledge on how to efficiently depolymerize biomass into fermentable sugars. We evaluated GH10 (Xyn10D and XT6) and GH11 (XynA and Xyn2A) β-xylanase performance alone and in combination with various GH family α-l-arabinofuranosidases (GH43 AXH-d and GH51 Abf51A) and α-d-glucuronidases (GH4 Agu4B and GH67 AguA) during xylan depolymerization. No synergistic enhancement in reducing sugar, xylose and glucuronic acid released from beechwood xylan was observed when xylanases were supplemented with either one of the glucuronidases, except between Xyn2A and AguA (1.1-fold reducing sugar increase). However, overall sugar release was significantly improved (≥1.1-fold reducing sugar increase) when xylanases were supplemented with either one of the arabinofuranosidases during wheat arabinoxylan degradation. Synergism appeared to result from the xylanases liberating xylo-oligomers, which are the preferred substrates of the terminal arabinofuranosyl-substituent debranching enzyme, Abf51A, allowing the exolytic β-xylosidase, SXA, to have access to the generated unbranched xylo-oligomers. Here, it was shown that arabinofuranosidases are key enzymes in the efficient saccharification of hetero-xylan into xylose. This study demonstrated that consideration of GH family affiliations of the carbohydrate-active enzymes (CAZymes) used to formulate synergistic enzyme cocktails is crucial for achieving efficient biomass saccharification.Item Open Access Drought tolerance responses in vegetable-type soybean involve a network of biochemical mechanisms at flowering and pod-filling stages(MDPI, 2021) Moloi, Makoena Joyce; Van der Merwe, RouxleneSevere drought stress affects the production of vegetable-type soybean (Glycine max L. Merrill), which is in infancy for Africa despite its huge nutritional benefits. This study was conducted under controlled environmental conditions to establish the effects of severe drought stress on ascorbate peroxidase (APX), guaiacol peroxidase (GPX), and glutathione reductase (GR) activities as well as proline, total soluble sugars (TSS), and hydrogen peroxide (H2O2) contents of five vegetable-type soybean cultivars (UVE8, UVE14, UVE17, AGS354, AGS429) at flowering and pod-filling stages. Drought induced significant increases in the contents of proline (selectively at pod filling for AGS429), TSS (at both stages for AGS429, and only at pod filling for UVE14), and malondialdehyde (AGS354 at flowering; UVE17 at pod filling). UVE8 and AGS354 had the highest H2O2 levels at flowering under drought stress, while AGS429 had the lowest. However, AGS429 was the only cultivar with significantly increased H2O2 under drought stress. Furthermore, drought stress induced significant increases in APX, GPX, and GR activities at flowering for AGS429. AGS354 recorded the highest decline for all antioxidative enzymes, while UVE17 decreased for GPX only. All biochemical parameters, except H2O2, were significantly higher at pod filling than at the flowering stage. The relationship between H2O2 and total seed mass (TSMP) or total seed per plant (TSP) was significantly positive for both stages, while that of TSS (at flowering) and proline (at pod filling) were significantly related to total pods per plant (TPP). The study suggests that during drought, the tolerance responses of vegetable-type soybean, APX, GPX, and GR (especially at the flowering stage), function in concert to minimize H2O2 production and lipid peroxidation, thereby allowing H2O2 to function in the signaling events leading to the induction of drought tolerance. The induction of TSS at flowering and proline at pod filling is important in the drought tolerance response of this crop.Item Open Access Solvent retention capacity and gluten protein composition of durum wheat flour as influenced by drought and heat stress(MDPI, 2021) Labuschagne, Maryke; Guzman, Carlos; Phakela, Keneuoe; Wentzel, Barend; Van Biljon, AngelineDrought and temperature stress can cause considerable gluten protein accumulation changes during grain-filling, resulting in variations in wheat quality. The contribution of functional polymeric components of flour to its overall functionality and quality can be measured using solvent retention capacity (SRC). The aim of this study was to determine the effect of moderate and severe drought and heat stress on SRC and swelling index of glutenin (SIG) in six durum wheat cultivars with the same glutenin subunit composition and its relation with gluten protein fractions from size exclusion high performance liquid chromatography. Distilled water, sodium carbonate and sucrose SRC reacted similarly to stress conditions, with moderate heat causing the lowest values. Lactic acid SRC and SIG reacted similarly, where severe heat stress highly significantly increased the values. SIG was significantly correlated with sodium dodecyl sulphate sedimentation (SDSS) and flour protein content (FPC) under all conditions. Lactic acid SRC was highly correlated with FPC under optimal and moderate heat stress and with SDSS under moderate drought and severe heat. SIG was negatively correlated with low molecular weight glutenins under optimal and drought conditions, and combined for all treatments. The relationship between SRC and gluten proteins was inconsistent under different stress conditions.Item Open Access Salicylic acid improves growth and physiological attributes and salt tolerance differentially in two bread wheat cultivars(MDPI, 2022) Abdi, Neila; Van Biljon, Angeline; Steyn, Chrisna; Labuschagne, Maryke TineAbiotic constraints such as salinity stress reduce cereal production. Salicylic acid is an elicitor of abiotic stress tolerance in plants. The aim of this study was to investigate the effects of salicylic acid on two bread wheat cultivars (SST806 and PAN3497) grown under salt stress (100 and 200 mM NaCl) in the presence and absence of 0.5 mM salicylic acid. The highest salt concentration (200 mM), in both PAN3497 and SST806, increased the days to germination and reduced the coleoptile and radicle dry weights. The shoot dry weight was reduced by 75 and 39%, root dry weight by 73 and 37%, spike number of both by 50%, spike weight by 73 and 54%, grain number by 62 and 15%, grain weight per spike by 80 and 45%, and 1000 grain weight by 9 and 29% for 200 and 100 mM NaCl, respectively. Salicylic acid in combination with 100 mM and 200 mM NaCl increased the shoot, root, and yield attributes. Salicylic acid increased the grain protein content, especially at 200 mM NaCl, and the increase was higher in SST806 than PAN3497. The macro-mineral concentration was markedly increased by an increase of NaCl. This was further increased by salicylic acid treatment for both SST806 and PAN3497. Regarding micro-minerals, Na was increased more than the other minerals in both cultivars. Mn, Zn, Fe, and Cu were increased under 100 mM and 200 Mm of salt, and salicylic acid application increased these elements further in both cultivars. These results suggested that salicylic acid application improved the salt tolerance of these two bread wheat cultivars.Item Open Access Identifying quality protein maize inbred lines for improved nutritional value of maize in Southern Africa(MDPI, 2022) Amegbor, Isaac; Van Biljon, Angeline; Shargie, Nemera; Tarekegne, Amsal; Labuschagne, MarykeMalnutrition, as a result of deficiency in essential nutrients in cereal food products and consumption of a poorly balanced diet, is a major challenge facing millions of people in developing countries. However, developing maize inbred lines that are high yielding with enhanced nutritional traits for hybrid development remains a challenge. This study evaluated 40 inbred lines: 26 quality protein maize (QPM) lines, nine non-QPM lines, and five checks (three QPM lines and two non-QPM lines) in four optimum environments in Zimbabwe and South Africa. The objective of the study was to identify good-quality QPM inbred lines for future hybrid breeding efforts in order to increase the nutritional value of maize. The QPM lines had a lower protein content (7% lower) than that of the non-QPM lines but had 1.9 times more tryptophan and double the quality index. The lysine- and tryptophan-poor y-zein protein fraction was 41% lower in QPM than in non-QPM, with a subsequent increase in y-zein. There was significant variation within the QPM inbred lines for all measured quality characteristics, indicating that the best lines can be selected from this material without a yield penalty. QPM lines that had both high protein and tryptophan levels, which can be used as parents for highly nutritious hybrids, were identified.Item Open Access The photosynthetic efficiency and carbohydrates responses of six edamame (Glycine max. L. Merrill) cultivars under drought stress(MDPI, 2022) Hlahla, Jeremiah M.; Mafa, Mpho S.; Van der Merwe, Rouxlene; Alexander, Orbett; Duvenhage, Mart-Mari; Kemp, Gabre; Moloi, Makoena J.Vegetable-type soybean, also known as edamame, was recently introduced to South Africa. However, there is lack of information on its responses to drought. The aim of this study was to investigate the photosynthetic efficiency and carbohydrates responses of six edamame cultivars under drought stress. Photosynthetic efficiency parameters, including chlorophyll fluorescence and stomatal conductance, were determined using non-invasive methods, while pigments were quantified spectrophotometrically. Non-structural carbohydrates were quantified using Megazyme kits. Structural carbohydrates were determined using Fourier-transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). Drought stress significantly increased the Fv/Fm and PIabs of AGS429 and UVE17 at pod filling stage. Chlorophyll-a, which was most sensitive to drought, was significantly reduced in AGS429 and UVE17, but chlorophyll-b was relatively stable in all cultivars, except UVE17, which showed a significant decline at flowering stage. AGS354 and AGS429 also showed reduced chlorophyll-b at pod filling. UVE17 showed a significant reduction in carotenoid content and a substantial reduction in stomatal conductance during pod filling. Drought stress during pod filling resulted in a significant increase in the contents of trehalose, sucrose and starch, but glucose was decreased. Chlorophyll-a positively correlated with starch. The FTIR and XRD results suggest that the cell wall of UVE14, followed by UVE8 and AGS429, was the most intact during drought stress. It was concluded that carotenoids, stomatal conductance, starch and hemicellulose could be used as physiological/biochemical indicators of drought tolerance in edamame. This information expands our knowledge of the drought defense responses in edamame, and it is essential for the physiological and biochemical screening of drought tolerance.Item Open Access Defining the frontiers of synergism between cellulolytic enzymes for improved hydrolysis of lignocellulosic feedstocks(MDPI, 2021) Mafa, Mpho S.; Pletschke, Brett I.; Malgas, SamkeloLignocellulose has economic potential as a bio-resource for the production of value-added products (VAPs) and biofuels. The commercialization of biofuels and VAPs requires efficient enzyme cocktail activities that can lower their costs. However, the basis of the synergism between enzymes that compose cellulolytic enzyme cocktails for depolymerizing lignocellulose is not understood. This review aims to address the degree of synergism (DS) thresholds between the cellulolytic enzymes and how this can be used in the formulation of effective cellulolytic enzyme cocktails. DS is a powerful tool that distinguishes between enzymes’ synergism and anti-synergism during the hydrolysis of biomass. It has been established that cellulases, or cellulases and lytic polysaccharide monooxygenases (LPMOs), always synergize during cellulose hydrolysis. However, recent evidence suggests that this is not always the case, as synergism depends on the specific mechanism of action of each enzyme in the combination. Additionally, expansins, nonenzymatic proteins responsible for loosening cell wall fibers, seem to also synergize with cellulases during biomass depolymerization. This review highlighted the following four key factors linked to DS: (1) a DS threshold at which the enzymes synergize and produce a higher product yield than their theoretical sum, (2) a DS threshold at which the enzymes display synergism, but not a higher product yield, (3) a DS threshold at which enzymes do not synergize, and (4) a DS threshold that displays anti-synergy. This review deconvolutes the DS concept for cellulolytic enzymes, to postulate an experimental design approach for achieving higher synergism and cellulose conversion yields.Item Open Access Fusarium casha sp. nov. and F. curculicola sp. nov. in the Fusarium fujikuroi species complex isolated from Amaranthus cruentus and three weevil species in South Africa(MDPI, 2021) Vermeulen, Marcele; Rothmann, Lisa A.; Swart, Wijnand J.; Gryzenhout, MariekaTrials are currently being conducted in South Africa to establish Amaranthus cruentus as a new pseudocereal crop. During recent surveys, Fusarium species were associated with weevil damage in A. cruentus fields. Preliminary studies showed that some of these Fusarium species grouped into two distinct clades within the F. fujikuroi species complex. The aim of this study was to characterize these isolates based on the morphology and phylogeny of the translation elongation factor 1 (TEF1 ) gene region, ß-tubulin 2 (ßT) gene region and RNA polymerase II subunit (RPB2), and to determine if these isolates are pathogenic to A. cruentus. Phylogenetic and morphological studies showed that these two clades represent two novel species described here as F. casha and F. curculicola. Both species were shown to have the potential to be pathogenic to A. cruentus during routine greenhouse inoculation tests. While isolations indicate a possible association between these two species and weevils, further research is needed to understand this association and the role of weevils in disease development involving F. casha and F. curculicola in A. cruentus.