Thesis (M.Sc. (Agronomy)) -- University of Limpopo, 2021 / Soybean (Glycine max L.) is one of the valuable leguminous crops with grain used for human consumption, livestock feeding, bio-fuel (bio-diesel) production, vegetable oil and is a protein resource. The crop also fixes atmospheric nitrogen. The study aimed to evaluate the performance of two soybean varieties to a combination of inoculation and seaweed extract. The research trial was conducted at Syferkuil farm during 2017/2018 summer season in which two soybean varieties (TGx 1937-1F promiscuous) and (PAN 1583R specific-nodulating) were evaluated for their growth, nodulation and yield responses to rhizobium inoculation and seaweed extract (0 % rate (0ml/12L), 50 % rate (30ml/12L) and 100 % (60ml/12L). The research trial was laid out as a split-split plot arrangement fitted in RCBD with four replications. Inoculation significantly influenced grain yield at P≤0.05, seed nutrient content and total above-ground biomass, except for seed potassium. No inoculation significantly achieved higher primary branches per plant at P≤0.05, pod number per plant, stem diameter (P≤0.05), grain yield (P≤0.05), harvest index and total above-ground biomass at P≤0.001. The variety TGx 1937-1F had significantly (P≤0.001) higher nodule number per plant, effective nodules per plant, nodule dry weight, dried shoot biomass, leaf number per plant, pod number per plant, primary branches per plant, stem diameter, plant height, leaf chlorophyll content, total above-ground biomass, grain yield and seed iron (Fe) content. Application of full rate seaweed extract significantly (P≤0.001) increased primary branches per plant, stem diameter, leaf number per plant, plant height, shelling percentage, total above-ground biomass, grain yield, and seed content of calcium (Ca), potassium, magnesium (Mg), manganese (Mn) and sodium (Na) all at P≤0.05. Seaweed extract rate at 0 % obtained the highest harvest index (P≤0.001). Inoculation and variety TGx 1937-1F interaction exhibited a significant increase on leaf number per plant at P≤0.001, primary branches per plant at P≤0.001 and plant height at P≤0.001. Variety TGx 1937-1F, without inoculation, obtained significantly higher pod number per plant (P≤0.001), stem diameter at P≤0.001, grain yield at P≤0.05 and total above-ground biomass at P≤0.001. Variety PAN 1583R, without inoculation, obtained significantly higher harvest index and shelling percentage at P≤0.001. Interaction of vi inoculation and seaweed extract showed that no inoculation × 100 % rate of seaweed extract significantly (P≤0.001) increased primary branches per plant, leaf number per plant, stem diameter, pod number per plant and plant height. Interaction of inoculation × 100 % rate of seaweed extract increased grain yield (P≤0.001) and total above-ground biomass at P≤0.001. Inoculation × 50 % rate of seaweed extract interaction increased shelling percentage at P≤0.001. No inoculation × 0 % rate of seaweed extract interaction obtained significantly higher harvest index (P≤0.001). Interaction of variety and seaweed extract showed that variety TGx 1937-1F × 100 % rate of seaweed extract significantly increased primary branches per plant (P≤0.001), pod number per plant at P≤0.001, grain yield at P≤0.001 and total above-ground biomass at P≤0.001. The variety TGx 1937-1F × 50 % rate of seaweed extract significantly raised the size of stem diameter (P≤0.01) and plant height (P≤0.001). Three-way interactive effects of inoculation × variety TGx 1937-1F × 100 % rate of seaweed extract obtained significantly higher number of shelling percentage at P≤0.001, leaf number per plant at P≤0.05 and primary branches per plant at P≤0.001. Interaction of no inoculation × variety TGx 1937-1F × 100 % rate of seaweed extract obtained significantly high pod number per plant at P≤0.001, grain yield at P≤0.05, total above-ground biomass at P≤0.001 and plant height at P≤0.001. No inoculation × PAN 1583R × 100 % rate of seaweed extract interaction had a higher harvest index at P≤0.001. The study showed that inoculation, seaweed extract, or their combination generally enhanced seed nutrient content, especially in variety TGx 1937-1F. The study further showed that promiscuous soybean (TGx 1937-1F) had higher grain yield, under stressful growing conditions as compared to PAN 1583R variety. This implies that soybean variety TGx 1937-1F, with 50 % or 100 % application rate of seaweed extract could be recommended to smallholder farmers. Key words: Soybean, inoculation, seaweed extract, phenological development, growth, nodulation, grain yield and seed nutrient content. / National Research Foundation (NRF)

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According to the American Heart Association, cardiovascular disease (CVD) is the leading cause of death in the U.S., representing about 20-30% of all deaths every year in the U.S. Major risk factors for developing CVD include high blood lipid and LDL-cholesterol levels. A large number of heart attacks and strokes could be prevented by controlling these factors through lifestyle modifications and diet interventions. Epidemiological evidence shows that consumption of dry or common beans (Phaseolus vulgaris L.) has positive effects on reducing blood LDL-cholesterol and lipid levels. These health benefits are mainly attributed to the high content of dietary fiber (DF) of beans, including soluble and insoluble DF (SDF and IDF). Some proposed mechanisms to explain the cholesterol and lipid-lowering effects of DF are related to the physico-chemical properties (e.g. viscosity) of DF, and involve binding to bile salts (BS) in the small intestinal to prevent BS re-absorption which further promote cholesterol catabolism and delay lipid digestion. Nevertheless, the precise mechanisms are not fully understood yet. In addition, cooking and processing operations, and in particular high-hydrostatic pressure (HHP) processing, can modify the composition, structure and functional properties of foods; however, whether HHP affects the ability of beans to interfere with different aspects of lipid digestion remains unknown. The overall goal of this research is to understand how common beans and HHP processing impact the ability of beans to bind BS and influence lipid digestion in vitro. The specific objectives are 1) to evaluate the effect of HHP treatments (and compared it with conventional cooking (HT)) on the thermo-rheological and functional properties of dry beans; 2) to identify the impact of major bean components on the in vitro BS-binding ability of beans, the role played by the bean matrix and how this is affected by HHP processing; 3) to investigate how bean (micro)structure and fiber fractions, as well as HHP processing of dry beans, influence lipid digestion in vitro. Results showed that HT caused complete starch gelatinization and protein denaturation of beans, while HHP treatments induced partial or no starch gelatinization and a lower degree of protein denaturation, which resulted in enhanced protein solubility and emulsifying activity/stability. It was observed that, while HT treatment reduced the capacity of bean flours to retain BS because of severe disruption of the bean cell wall integrity, protein matrices, and starch granules, HHP treatments maintained or enhanced BS retention, possibly by promoting the formation of starch/protein/fiber networks able to entrap BS. Furthermore, by using an in vitro dialysis-based digestion model combined with viscosity measurements and thermal analysis, it was shown that the interaction between bean tissue materials and primary BS was not only related to viscosity but also involved hydrophobic linkages. The contribution of IDF and proteins (other than SDF) to retain BS was also significant. There was a different binding preference of beans to four primary BS with sodium glycochenodeoxycholate, the more hydrophobic BS, showing the largest retention levels while sodium taurocholate being the least effectively retained BS by beans. Diverse sequences of the same processing operations showed distinct impacts on BS-retention by dry beans. By means of an in vitro digestion model simulating conditions in the upper gastrointestinal tract, bean flours delayed the digestion of extrinsic lipids to a higher extent, compared to isolated IDF and SDF. Furthermore, HHP treatment and less severe mechanical disintegration maintained the ability of beans to modulate lipid digestion, which suggests the importance of bean structural integrity in reducing the lipolysis rate and extent by beans. Overall, this research work shows that HHP processing is a promising minimal processing technology to produce bean flours with improved functionality. It also highlights the importance of considering the structure of foods, and not just their nutrient content, when evaluating potential health impacts. This knowledge could be applied to develop a range of bean-based ingredients and formulations with desirable health benefits. This work can be extended to research the influence of beans on the gut microbiota and profile of secondary BS and short-chain fatty acids, which are also closely related to cholesterol and lipid metabolism. / Doctor of Philosophy / According to the American Heart Association, cardiovascular disease (CVD) is the leading cause of death in the U.S., representing about 20-30% of all deaths every year in the U.S. Around the world, millions of people are struggling to control the risk of CVD. Major risk factors for developing CVD include high blood lipid and LDL-cholesterol levels. A large number of heart attacks and strokes can be prevented by controlling the major risk factors through lifestyle modifications and diet interventions. Epidemiological evidence shows that consumption of dry beans (Phaseolus vulgaris L.) has positive effects on reducing blood LDL-cholesterol and lipid levels. These health benefits are mainly attributed to the high content of dietary fiber (DF) in beans. DF is carbohydrate polymers that are not hydrolyzed by the endogenous enzymes in humans. However, some of them (water-soluble DF) could increase viscosity and retain the absorption of bile salts (BS) in the small intestinal. The BS retention or the binding of BS could promote more cholesterol convert to BS (thus reduce cholesterol levels) and decrease lipid digestion. Therefore, due to the increased viscosity and BS retention ability of DF, dry beans could help to reduce the blood cholesterol and lipid levels and further help to prevent CVD. Moreover, different cooking and processing method could also affect the composition, microstructure and functional properties of foods. The purpose of this research was to determine how common beans and high hydrostatic pressure (HHP) (compared with hydrothermal (HT)) processing, a non-thermal processing, influence the ability of dry beans to retain bile salts and modulate lipid digestion in vitro. This study showed that severe HT treatment disrupted the bean cell wall integrity severally and reduced the BS retaining the efficiency of dry beans, while HHP treatment, produced minimally processed beans, improved the application properties of dry beans and maintained/enhanced BS-retention by dry beans. It also showed that the whole bean matrix (other than soluble DF) also contributes to retain BS and modulate lipid digestion, indicating the importance of retaining intact food structures. The integrity of bean structures through HHP treatment and less severe mechanical treatment could help to retain the ability of dry beans to reduce lipid digestion. These findings suggest that dry beans, with a high content of dietary fiber and resistant starch, have significant health benefits related to lowering cholesterol and lipid levels. Increasing the consumption of dry beans would definitely help to improve overall health. HHP, as a non-thermal processing technology, showed the potential to produce minimally processed bean products with enhanced health benefits and diverse application properties. This study could be extended through continuing research into the influence of beans on the gut microbiota, which are also closely related to the cholesterol and lipid metabolism regulation.

Edamame is becoming more popular in the U.S. due to its high nutritional value and potential health benefits. However, more than 70% of edamame is imported from outside of the U.S. Therefore, developing elite edamame genotypes is critically desirable to increase the domestic production of edamame in the U.S. Genotype, planting location, and harvest time play essential roles in the chemical composition of edamame, which further decide edamame's nutritional value and sensory characteristics. Therefore, the first goal of this study is to comprehensively evaluate the chemical composition of edamame genotypes grown in different locations. Ten selected edamame genotypes were grown in three locations in the U.S. - Whitethorne, Virginia (VA), Little Rock, Arkansas (AR) and Painter, VA. Sugars, alanine, protein, oil, neutral detergent fiber (NDF), starch, ash, and moisture contents, were comprehensively analyzed. The results showed that location had significant effects on all chemical components of edamame with p < 0.05. Compared to Painter and Little Rock, genotypes planted in Whitethorne had higher averaged free sucrose, fructose, glucose, raffinose, stachyose, and starch contents and total sweetness. The highest crude protein and oil contents were found on edamame planted in Painter, while Little Rock produced edamame with the highest free alanine, ash, and moisture contents. Genotype significantly affected chemical compositions except for NDF and raffinose. Therefore, planting location and edamame genotype should be considered when producing elite edamame for the U.S. market. Chemical composition changes with the development of edamame; therefore, harvest time is essential for harvesting high-quality edamame. The second objective of this study is to quantify the changes in both physical and chemical properties of edamame over bean development and apply a combined spectroscopy and machine learning (ML) technique to help determine the optimal harvest time. Physical and chemical properties were analyzed for edamame harvested at R5 (beginning seed), R6 (full seed), and R7 (beginning maturity) growth stages, and the spectral reflectance (360 – 740 nm) of edamame pods was measured using a handheld spectrophotometer. The samples harvested at different stages were labeled as 'early,' 'ready,' and 'late.' At R6, pod/bean weight and pod thickness reached the peak and then stayed stable, while sugar, alanine, starch, and glycine also peaked at R6 but declined afterward. The spectra-based ML method had high accuracy (0.95) when classifying 'early' and 'late' edamame, and the accuracy was 0.87 for classifying 'early' and 'ready' edamame. These results indicated that this spectra-based ML method could determine the optimal harvest time of edamame. Food waste and loss not only lead to economic loss but also significant greenhouse gas emissions. With edamame food/snack production increasing, edamame shells, the low-value byproduct from this processing, will potentially threaten the environment. Similar to other food processing byproducts, edamame shell is rich in dietary fiber (DF). However, the high concentration of insoluble dietary fiber (IDF) limits its application as a food additive. Therefore, extraction/modification processes are needed to convert IDF to soluble dietary fiber (SDF) and improve the properties of edamame shell-derived DF. Ball milling is one of the most efficient techniques to break down biomaterials into sub-micro-level particles. Citric acid, as a natural and safe food additive, can help break down cell walls and improve the dissolution of SDF by ionizing the hydrogen ions with carboxyl groups. Therefore, the third objective of this study is to develop a process that combines ball milling and citric acid treatments to produce SDF from edamame shells. We investigated different treatment parameters, including different citric acid concentrations, treatment temperatures and time, and the application of ball milling. To determine if the combined treatment can potentially improve the properties of the produced SDFs, we characterized the physicochemical, morphological, structural, rheological, thermal, and functional properties of SDFs produced at different conditions. The results showed that the highest SDF yield (19.5%) was found when the edamame shells were pretreated by a ball mill. In addition, the combined citric acid and ball milling treatment altered several properties of the produced SDFs, including particle size, morphology, and crystallinity. Moreover, ball milling treatment led to a higher exothermic temperature peak of SDF indicating better thermal stability. All produced SDFs significantly elevated the production of short-chain fatty acids during in vitro fermentation (compared to the control fermentation) which indicated their potential benefits of promoting gut health. Overall, we demonstrated that ball-milling-assisted citric acid processing can be an effective green technique to produce SDF from edamame shells. The SDF produced from edamame shells can be regarded as a promising and novel ingredient with great potential to be used in foods. / Doctor of Philosophy / Edamame is becoming increasingly popular among consumers in the U.S. because it is nutritious and good for health. However, more than 70% of edamame in the U.S. market is imported from other countries. Therefore, having more edamame genotypes that adapt to the growing environment in the U.S. will help increase the domestic production of edamame. Genotype and planting location are essential in deciding edamame's nutritional value and taste. Therefore, the first objective of this study is to comprehensively understand the nutritional value of different edamame genotypes grown in three planting locations. The results showed that both location and genotype affected the nutritional values of edamame, indicating that planting location and edamame genotype should be considered when developing better edamame for the U.S. market. Nutritional value and sweetness change with the growth of edamame beans. Therefore, harvest time is crucial for harvesting edamame with better nutrition and taste. This study's second objective is to observe edamame's nutritional factors and sweetness over bean development and develop a method using a handheld colorimeter to help determine the optimal harvest time. The results showed that the edamame harvested at the full seed stage (called R6) is the sweetest compared to the other two stages. In addition, the handheld colorimeter combined with the machine learning technique showed high accuracy in separating 'early' and 'late' harvested edamame and 'early' and 'ready' harvested samples. These results indicated that the combination of colorimeter and machine learning could help determine the optimal harvest time of edamame. Food waste and loss not only lead to economic loss but also significant greenhouse gas emissions. Edamame shells, the low-value byproduct from edamame snack/food processing, will potentially threaten the environment if edamame consumption keeps increasing. Like other food waste, edamame shell is rich in dietary fiber (DF). Therefore, it is vital to find a way to recover the DF in edamame for other applications. Ball milling is a green technology that can efficiently break down big particles. Citric acid is a natural and safe food additive and can help break down insoluble cell walls. Therefore, this study aims to produce soluble dietary fiber (SDF) from edamame shells using ball milling and citric acid. We proved that ball-milling assisted acid processing can be an environmentally friendly method to produce edamame shell SDF which can potentially be used as a suitable food ingredient.

Fusarium root rot (FRR) disease, caused by the fungus Fusarium solani f. sp. phaseoli (FSP), is an important soil-borne disease reducing common bean (Phaseolus vulgaris L.) yields, and hence food security, in Uganda and elsewhere in developing countries where the crop is grown without fungicides. The key aim of this study was to elucidate the significance of bean root rot (BRR), appraise methods for screening germplasm for resistance to FRR, determine the genotypic variability of resistance, and the inheritance of resistance to FRR in common bean. This information was deemed useful in devising an appropriate strategy for breeding FRR resistance in beans. A participatory rural appraisal (PRA) was conducted in south-western and eastern Uganda to ascertain farmers’ awareness of BRR and their influence on preferred bean varieties. Bean root rot is considered to be the most devastating and most recognised disease, especially in south-western Uganda. Control measures for BRR were very minimal, and in some cases, non-existent. Use of resistant varieties to control the disease was not evident, because the most popular varieties were susceptible to the disease. The resistant bean varieties currently available have undesirable characteristics such as small seed size, black seed and late maturity. Large-seeded bean varieties, even though cited as being more susceptible to BRR than the small-seeded varieties, are still very popular. The study highlighted the need for breeding FRR resistance in the large-seeded bean varieties that are highly preferred by farmers. Four isolates of FSP (FSP-1, FSP-2, FSP-3 and FSP-4) were tested for pathogenicity under screenhouse and laboratory conditions. In addition, three methods of storing and maintaining the viability of FSP isolates were appraised. The isolate FSP-3, was found to be the most pathogenic, resulting in 100% disease incidence on all bean varieties tested, with high severity scores. The potato dextrose agar (PDA) slants stored at 5oC were found to be the best method of storage for pathogenic isolates. The FSP-3 isolate was subsequently utilised for screening bean lines for resistance to FRR. The influence of soil composition, irrigation frequency, and inoculation technique on the severity of FRR was studied on six bean lines. Interactions of irrigation frequency, soil composition, and bean lines were not significant. The 50% swamp soil:50% forest soil composition and forest soil alone categorized the varieties most distinctly according to their reaction to FRR. Also, the best distinct classification for the varieties was obtained under treatments that were watered daily and once in a week. Based on economic considerations, the standard forest soil and daily irrigation were subsequently adopted for screening bean germplasm for resistance to FRR. It was also found that sorghum seed as a medium for pathogen inoculation was better than the agar slurry medium. One hundred and forty seven common bean varieties were evaluated for resistance to FRR (isolate FSP-3) under screenhouse conditions. In order to confirm this resistance, 46 common bean lines selected from the screenhouse trial were further evaluated using natural inoculum in a BRR-infested field. Forty-four varieties comprising ten large-seeded, four medium-seeded and 30 small-seeded varieties showed moderate resistance to FRR; but none were resistant or immune to the disease. Based on adaptability, eight moderately resistant varieties were selected for use as parents in the study of inheritance of resistance to FRR. A 12 x 12 diallel mating design was utilised to develop 66 F1 and F2 populations, plus their reciprocal crosses, with the aim of studying the mode of inheritance of resistance to FRR. The F1 and F2 progeny evaluations showed that FRR resistance was mainly governed by additive genes in most populations. However, there were a few crosses which displayed highly significant specific combining ability (SCA) effects, implying that dominant effects were important in some populations. Maternal effects were also highly significant at both the F1 and F2 generations, suggesting that resistance was modified by cytoplasmic genes. The non-maternal effects were also significant in some populations, suggesting that the cytoplasmic genes were interacting with nuclear genes. The number of genes governing resistance to FRR varied from two to nine among the eight sources of resistance. The allelism test of resistant x resistant populations, and the observation of continuous distributions of severity scores, suggested the presence of many loci governing FRR resistance in beans. Broad sense heritability of disease resistance varied from 0.22-0.69, while heritability in the narrow sense was estimated as 0.35-0.49 in the populations. These results suggested that selection and backcrossing to both parents would be the best breeding procedures for improving resistance in the popular large-seeded bean varieties in Uganda. However, there could be complications in breeding for resistance to FRR in beans, because resistance was modified by cytoplasmic gene effects and their interaction with nuclear genes in some of the populations. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.