Master of Science / Food Science Institute / Fadi M. Aramouni / With an increasing number of people with celiac disease, the need for gluten-free products is inevitable. Sorghum is a grain safe for celiac patients. Therefore, the purpose of this work was to characterize four sorghum hybrids in terms of their grain and flour; then utilize the hybrids in a wheat-free product and test for physical, chemical, textural, and sensory differences. Flour tortillas were chosen for their current popularity and the lack of research and availability for gluten-free flour tortillas. Grain characterization included diameter, weight, and hardness as measured by the Single Kernel Characterization System and Tangential Abrasive Dehulling Device. Flour characterization included flour and starch particle size distributions, total starch, amylose content, starch pasting properties, moisture, crude protein, and ash content. Significant differences were found (p<0.05) among hybrids for each test except total starch. Gluten-free flour tortillas were made from the four sorghum hybrids in addition to a commercial sorghum flour. Tortilla weight, diameter, thickness, color, pH, Aw, and moisture content were measured along with extensibility and stretchability. A descriptive panel was trained and used to analyze the five samples. Significant differences were found (p<0.05) among samples for color, pH, Aw, and moisture content. Significant differences were also found (p<0.05) among samples for extensibility and stretchability. Extensibility was a more effective test in studying quality. The sensory panel found significant differences (p<0.05) for grain specks, angle of bend, rancidity, sweetness, springiness, hardness, and grittiness. The commercial flour had the highest score for angle bend and springiness and was, therefore, utilized in a consumer study. When compared to a gluten-free wrap already in the market, the sorghum flour tortilla made from this study scored significantly higher in all attributes, including overall acceptability. The commercial flour is thought to have preformed better than the other four samples due to its smaller particle size and greater starch damage allowing an increase in water absorption.

Marama bean (Tylosema esculentum (Burch) A. Schreib) is an underutilised, drought- tolerant legume native to the semi-arid areas of Botswana, Namibia and South Africa. The edible seeds of marama beans are good sources of protein and fat, and its flours can potentially enhance the nutritional quality of sorghum porridge, a staple in many African countries. Sorghum has a low protein content and is deficient in lysine. Compositing sorghum with lysine-rich marama flour when preparing porridges would address this problem. Utilisation of marama flour in composite porridge depends on sensory acceptance. In this study nutritional and sensory quality of marama-sorghum composite flours and porridges are investigated. Full fat and defatted flours from unheated and dry heated (150 °C /20 min) marama beans were composited with condensed tannin-free sorghum meal (30:70). Marama flours, sorghum meal, marama-sorghum composite flours and porridges were analysed for proximate composition, amino acid composition, energy value, fatty acid composition, total phenolic content and antioxidant activity. Porridges (10% solids) were subjected to descriptive sensory analysis, consumer acceptance test (n=52), texture analysis, pasting profiles and colour measurements. Sorghum porridge and porridge composite with full fat marama flour from heated beans were more acceptable than other composite porridges. Compositing sorghum meal with marama flour reduced the cooked sorghum aroma/flavour intensities prominent in sorghum porridge. Dry heating marama beans prior to milling resulted in a roasted nutty flavour in the final product. Porridges from heated marama beans were darker in colour than porridges from unheated beans. This could be attributed to flavour and colour development due to Maillard reaction during dry heating. Composite porridges from full fat marama flours were described as buttery/creamy. A bitter taste and aftertaste was perceived in porridges from defatted flours. Presence of fat seems to mask bitterness in porridges. Marama-sorghum composites and Tsabotlhe porridges were less viscous than sorghum porridge. This was confirmed by pasting profiles (low final viscosities) and texture analysis indicating reduced firmness. Starch contributes greatly to the textural properties of sorghum porridge. Marama flour has no starch. Compositing the flour with sorghum reduced starch levels in the composite porridges. This explains the reduction in viscosity and firmness of the porridges. Compositing sorghum meal with marama flours significantly increased protein and fat contents in flours and porridges. The fat content in marama-sorghum flours was in the range of 5.6 to 14.6%. Protein increase ranged from 61% to 96% in marama-sorghum composite porridges compared with sorghum porridge. There was 11-24% energy value increase in marama-sorghum composite porridges compared with sorghum porridge. Porridges composited with full fat flours contributed to higher energy values than those composited with defatted flours. This is because of fat energy contribution. Lysine content in marama flour was significantly higher than sorghum meal. Lysine content was 3 to 4 times higher in marama-sorghum porridges than in sorghum porridge. Unsaturated fatty acids were the most present in both marama-sorghum composite porridges and sorghum porridge. There was an increase in oleic acid as marama flour was composited with sorghum. Marama flours had higher levels of total phenolics than sorghum flour. Marama flour has high levels of tyrosine which could react with the Folin Ciocalteu reagent thus leading to an increase in measurable phenols. Marama flours from dry heated beans had higher total phenolic content and antioxidant activity than flours from unheated beans. Heating may have increased phenolic compound extractability due to release of bound phenolics. There was a positive correlation between the total phenolic content and antioxidant activity (r = 0.80, p < 0.05) in all samples. Findings from this study indicate that marama flour can potentially be used to improve the nutritional quality of sorghum porridges. Furthermore this study shows that use of full fat flour in marama-sorghum composite porridges give more acceptable sensory properties. / Dissertation (MScAgric)--University of Pretoria, 2011. / Food Science / unrestricted

Please read the abstract in the 00front part of this document Copyright 2004, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. Please cite as follows: Bayu, W 2004, Growth, development and yield responses of sorghum to water deficit stress, nitrogen fertilizer, organic fertilizer, and planting density, PhD thesis, University of Pretoria, Pretoria, viewed yymmdd < http://upetd.up.ac.za/thesis/available/etd-09202006-093510 / > / Thesis (PhD (Agronomy))--University of Pretoria, 2004. / Plant Production and Soil Science / unrestricted

Von 2005 bis 2011 wurden auf unterschiedlichen Versuchsstandorten in Deutschland die Trockenmasseerträge von Sorghumhirsesorten und Maissorten verglichen. Es stellte sich heraus, dass der Anbau von Sorghumhirsen auf allen geprüften Standorten mit Erfolg möglich ist und eine Alternative zum Mais im Energiefruchtfolgesystem darstellt. Die Sorghum bicolor-Sorten erreichen ein höheres Ertragspotenzial als die Hybridsorten (Sorghum bicolor x sudanense). Die Hybridsorten zeichnen sich durch eine schnellere Abreife aus und erreichen silierfähige Trockensubstanzgehalte. Verglichen mit Mais erzielen die Sorten der Sorghumart Sorghum bicolor insbesondere auf den D-Süd-Standorten gleiche oder bessere Erträge.

Ausgewertet wurde der Anbau von Sorghumhirsen auf Versuchsfeldern unterschiedlicher bodenklimatischer Bedingungen. Dabei variierten Versuchsparameter wie Bodenbearbeitung, Aussaattermin, Saatstärke, Reihenweite, Saattiefe und Unkrautregulierung. Für Sorghum bicolor wurde eine optimale Saatstärke von 25 und für die Sudangräser von 40 Körnern/m² ermittelt. Die Reihenweiten sind von untergeordneter Bedeutung, sodass die übliche Drillweite im Betrieb nicht verändert werden muss. Die Versuche zeigen, dass Sorghum sowohl als Drill-, Einzelkorndrillsaat als auch im Mulchsaatverfahren ausgebracht werden kann. Heft 24/2012 der Schriftenreihe des Landesamtes für Umwelt, Landwirtschaft und Geologie mit dem Titel „Ergebnisse mehrjähriger Sortenversuche Sorghumhirsen“ bilanziert die Trockenmasseerträge von Sorghumhirsesorten im Vergleich zu Mais.

Thesis (Ph.D.(Microbiology)) -- University of Limpopo, 2022 / Traditional sorghum juice is produced in many African countries for human consumption. The juice is very rich in calories, B-group vitamins including thiamine, folic acid, riboflavin, nicotinic acid, and essential amino acids such as lysine. Low earning income women at village level produce sorghum juice for home consumption and sale. The short shelf life (2 to 3 days) of sorghum juice is a major problem for both the brewers and consumers of this drink. The aim of the study was to use 12 Combretum plants to improve the microbiological quality and shelf life of sorghum juice. Fresh stems and leaves of C. caffrum, C. vendae, C. erythrophyllum, C. elaegnoides, C. apiculatum, C. imberbe, C. adenogdium, C. padoides, C. bracteosum, C. kraussii, C. mkuzense and C. zeyherii were collected at Nelspruit, National Botanical Gardens, Mpumalanga, South Africa. Voucher specimens and tree labels were used to verify the identity of the plants. The stems and bark collected were cut into pieces and air dried for 30 days. When dried, the plant material was ground to a fine powder and stored in paper bags at room temperature. The wood was burnt in an open fire; fuel was not used to minimise contamination. The qualitative phytochemical composition of both the leaves and stems of Combretum plants analysed in this study revealed the presence of saponins, tannins, terpenoids, steroids, cardiac glycosides and flavonoids. The following phytoconstituents were lost in the ashes; tannins with the exception of C. mkuzense and C. padoides; cardiac glycosides and flavonoids. The quantitative phytochemical analyses revealed that both the leaves, stems and some ashes such as C. apiculatum and C. vendae contained appreciable levels of phenolic compounds, tannins and flavonoids. Quantitative analysis of antioxidant activity, the 2, 2, diphenyl-1-picrylhydrazyl (DPPH) assay was used as a screen test for the radical scavenging ability of the compounds present in the different 36 70% acetone extracts. DPPH screening method indicated great scavenging activity with the 70% acetone leaf extracts of C. kraussii, C. zeyherii and C. mkuzense. The leaf and stem extracts showed substantial great antioxidant activity in a concentration-dependent manner. There was a significant decrease in the antioxidant activity in the ashes (p=001), when compared to both the leaves and the stems. The proximate and nutritional analysis of the 70% acetone extracts were performed by AOAC and ICPE protocols, respectively. The results indicated that all the extracts had substantial amounts of ash, moisture, protein and energy. Mineral content of the plant parts was analysed as well, calcium had the highest concentration, while zinc was lowest in concentration. The mineral content decreased significantly in the stems (p ≤ 0.05) when compared to the leaves. There was a further decrease in mineral content with regard to the ashes with the exception of calcium. Based on these findings, the leaves and ashes of C. adenogonium and C. apiculatum could provide a good source of calcium in the diet, while C. adenogonium, C. bracteosum and C. apiculatum had high levels of sodium. A serial micro-dilution assay was used to determine the minimum inhibitory concentration (MIC) values for 70% plant extracts using tetrazolium violet reduction as an indicator of growth. Two Gram-positive (Stapylococcus aureus ATCC 29213 and Enterobacter faecalis ATCC 29212) and two Gram-negative (Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853) bacterial strains were used in this study. The leaves had good antibacterial properties with the lowest MIC value being 0.04 mg/ml against E. coli and S. aureus. E. faecalis was found to be resistant against all the leaves with the exception of C. imberbe. The stem extracts of Combretum spp. tested in the study showed antimicrobial properties with the lowest MIC value being 0.04 mg/ml against E. coli shown by C. bracteosum. However, E. faecalis was resistant against all the 12 plants tested. All the test microorganisms showed resistance to the ashes, with the exception of S. aureus, which was found to be susceptible to 75% of the test ash extracts with the lowest MIC value of 0.16 mg/ml. Cytotoxicity and anticancer activity of the acetone extracts of the 12 Combretum plants were evaluated using tetrazolium-based colorimetric assay (MTT assay) on A549 lung carcinoma cells. The assays revealed that 50% of the leaf extracts of tested plants showed cytotoxicity and cell proliferation inhibition in A549 lung carcinoma cells in a concentration-dependent manner. The A549 cells were more sensitive to the following plants: C. elaegnoides, C. erythrophyllum C. imberbe, C. kraussii and C. mkuzense. The following stems extract, C. adenogdium and C. caffrum did not have any anticancer activity, whereas C. apiculatum and C. bracteosum were only able to reduce cell viability to less than 60%. C. mkuzense, C. padoides, C. vendae and C. zeyherii acted in a concentration-depended manner with the greatest activity seen at the highest concentration (1000 µg/ml). The plants had activity at concentrations between 31.25 and 1000 µg/ml allowing only 20% and 50%, respectively, of the cells to remain viable. Ashes from C. mkuzense showed good anti-cancer activity at the highest concentration (1000 µg/ml) reducing cell viability to around 10%. Enterobacteriaceae, total coliform, S. aureus, B. cereus, E. coli and lactic acid bacteria viability were studied during the four weeks storage period of prepared sorghum juices. Juice samples were collected after preparation; the samples were serially diluted using peptone water. Tempo instrument (Biomereiux) was used to enumerate total coliform, total aerobic count, E. coli, S. aureus, lactic acid bacteria, enteric bacteria, yeast and mould using the most probable number following the manufacturers instruction. C. mkuzense and C. padoides plants were able to inhibit the growth of B. cereus, lactic acid bacteria and S. aureus during the first three weeks of storage. E. coli was not present throughout the four weeks storage time. Vitek 2 Compact (Biomereiux) was used for the characterisation and identification of the dominant bacterial isolates using biochemical reactions. The isolates were characterised by morphological differences. Sixty five percent of the isolates were the Enterobacter genus that are commonly found in soil, water, and sewage. The nutritional composition and sensory properties of the prepared sorghum juices treated with Combretum plants were investigated. All the juices had appreciable amounts of protein, ash and energy. Sorghum juice treated with ashes of C. caffrum, C. erythrophyllum and C. kraussii had the highest levels of proteins when compared with other treated sorghum juices. The sorghum juices prepared in the study had varying levels of trace element or minerals with potassium -(3, 55 – 104 mg/l) and calcium (3.2-148 mg/l). Similarly, cobalt (-1.22), coppr (-0.99), iron(-0.962), magnesium (0.004), sodium (-0.145), nickel (-2.7)) and zinc (-1.2)are present in very low amounts. The juices treated with ashes had relatively higher levels of calcium, potassium, magnesium and sodium. Sorghum juices treated with the ashes had better sensory and organoleptic properties when compared with those treated with the leaves. The juices treated with the ashes of C. caffrum and C. bracteseum were more accepted by most of the panellist when compared with other treated juices. Overall, this study presents valuable information on the phytochemical composition, nutritional composition and antioxidant properties of some Combretum species in South Africa. It recommended its use as food and in pharmaceutical preparations for the local industries. In addition, Combretum plants showing the effects tested in this study may be explored further for development into drugs. functional food as food preservatives and nutraceutical applications, beside their traditional use / University of Limpopo and Foodbev SETA

Thesis (Ph.D. (Plant Production)) -- University of Limpopo, 2022 / Sustainable food production has been a major challenge in the era of climate change and a growing population in the twenty-first century. However, climate change scenarios such as extreme temperatures and fluctuations in annual precipitation continue to pose a great threat to agricultural production systems. On the other hand, anthropogenic activities such as conventional farming continue to contribute to climate change through the emission of greenhouse gases while not sustaining agricultural production. The Food and Agriculture Organization of the United Nations (FAO-UN) developed the concept of Climate-Smart Agricultural (CSA) production with the idea of securing food in the face of global change. No-tillage and intercropping systems are among the traditional practices that are advocated as components of climate-smart traditional practices, especially in the semi-arid regions of Africa like the Limpopo Province. Producing sorghum and cowpeas using CSA practices such as intercropping under no-tillage is envisaged to increase productivity and soil fertility under Limpopo Province's dryland conditions. However, there is still limited information on how grain sorghum-cowpea intercrop will respond in terms of growth, physiological productivity, and carbon dioxide emissions in the system, especially under no-tillage and different growing conditions. Furthermore, more field data is required for predictions of future scenarios using simulating crop models such as the Agricultural Production system sImulator (APSIM). Hence, a no-till Randomized Complete Block Design (RCBD) in a 2 x 4 x 2 factorial arrangement was conducted at two locations (Syferkuil and Ofcolaco) in the Limpopo Province during the 2018/19 and 2020/21 cropping seasons to generate data on sorghum and cowpea growth, physiology, productivity as well as carbon dynamics under planted and simulated intercropping system. Leaf gaseous exchange and leaf area index (LAI) were measured on fully developed grain sorghum and cowpea leaves in both the binary and sole cultures of sorghum and cowpea. The CO2 measurements were taken from each plot using a GMP343 CO2 probe along with an MI70 data logger. Aboveground biomass was collected for each crop from two plants at vegetative, flowering, physiological and harvest maturity and oven-dried at 65 oC for 48 hours. In the 2020/21 cropping season, cowpea at Ofcolaco failed to produce grain. Hence, only the grain yield of the 2018/19 cropping season from Ofcolaco is presented in this thesis. Grains collected for each crop from a 2.7m2 area were taken to the laboratory to determine grain yield and yield components. Harvest index (HI) and land equivalent ratio (LER) for each crop were also determined. In the laboratory, the total nitrogen (%) and natural abundance of 15N (δ15N‰) were determined using an isotope ratio mass spectrometer with an N analyzer. Growth (biomass) and yield (grain) data obtained from APSIM were compared with data collected from a two-year field experiment at Syferkuil. Multi-variate analysis of variance (ANOVA) model to fit each response variable using the Statistical Analysis System (21 SAS version 9.4). Mean separation was done where the means were different using the least significant difference (LSD) at probability levels of p ≤ 0.05. Intercropping system and the density of the companion crop cowpea had a significant (p ≤ 0.05) effect on the physiological responses of sorghum and cowpea, cowpea yield and yield components at the two experimental sites across seasons. However, grain yield and yield components of sorghum were not affected by intercropping or the density of cowpea. Only cultivars of sorghum were significantly different for grain yield and yield components. At Syferkuil, Enforcer produced the highest grain yield of 4338 kg ha-1 in 2018/19, while NS5511 accumulated the highest grain yield of 2120 kg ha-1 during the 2020/21 cropping seasons. At Ofcolaco, Enforcer and Avenger were observed to be relatively high-yielding cultivars with a mean grain yield of 2625 kg ha- 1 and 1191 kg ha-1 during the 2018/19 and 2020/21 cropping seasons, respectively. In the 2018/19 and 2020/21 cropping seasons, respectively, cowpea accumulated about 93% and 77% more grain yield in sole compared to binary culture. At Ofcolaco, about 96% more grain yield was obtained in sole compared to binary cultures during the 2018/19 cropping season. Furthermore, cowpea accumulated over 55% and 49% of grain yield when grown at high compared to low population density at Syferkuil and Ofcolaco, respectively. The investigation on the impact of the intercropping system on CO2 emissions and soil carbon stocks revealed that in 2018/19 at Syferkuil and 2020/21 at Ofcolaco, intercropping systems emitted 11% and 19% less CO2 respectively than the sole cropping systems. In both diverse agro-ecological sites, low cowpea density consistently resulted in higher CO2 emissions than high density. The sorghum-cowpea intercropping system significantly influenced the biological nitrogen fixation of cowpea. Intercropping was found to improve the biological nitrogen fixation of cowpea if a density of 74074 plants ha-1 is used. The APSIM model was able to capture the dynamics of biomass and grain yields in the sole and intercropping system under different densities of cowpea. The findings of this study revealed some useful insights. Firstly, biomass accumulation depended on the cultivar in intercrop as well as the density of cowpea. Secondly, cowpea at a density of 74074 plants ha1 was found to be a good crop to intercrop with grain sorghum as it did not show any significant variation in terms of grain yield and yield components of sorghum. The sorghum cultivar, Enforcer and NS5511 were the best performing cultivars in terms of grain yields at Syferkuil and Ofcolaco. Thirdly, the intercropping system under high cowpea density reduced CO2 emission rates while improving soil nitrogen (N) and carbon stocks. Based on the results of this study, grain sorghum-cowpea intercrop can be adopted as a component of a climate-smart practice to improve crop growth, physiology, as well as productivity compared to sole cropping. However, the grain sorghum cultivar and the density of cowpea should be taken into consideration as they affect the productivity of the two crops. The two seasons data generated from this study was useful in simulating the productivity of intercropping practice using APSIM. However, more field and weather data is required to run long-term simulations on intercropping as a component of the climate-smart method using crop modelling techniques. / National Research Foundation (NRF), Departments of Science and Innovation (DSI) and VLIR-IUC (Belgium)

Industry demand for local sources of grain for animal feed has increased sorghum production in the mid-Atlantic region of the U.S. Sorghum anthracnose (causal agent Colletotrichum sublineola) and the grain mold complex, which includes mycotoxin-producing Fusarium spp., limit the yield and quality of grain sorghum in humid climates worldwide. A majority of U.S. grain sorghum production is in arid regions, and management strategies have not been developed for the mid-Atlantic U.S. where warm, wet conditions favor disease. The specific objectives of this research were to: (1) determine the effectiveness of fungicides and their application timing for the management of sorghum foliar anthracnose, (2) compare five grain sorghum hybrids for their susceptibility to foliar anthracnose, grain mold and mycotoxin contamination under field conditions, (3) integrate host resistance and fungicide application to manage anthracnose and grain mold, and (4) identify Fusarium spp. associated with grain mold and mycotoxin contamination of sorghum in the mid-Atlantic U.S. For Objective 1, it was determined that a single application of pyraclostrobin-containing fungicide no later than flowering reduced anthrancose, protected yield and maximized farm income. Objective 2 focused on sorghum hybrid selection as a disease management tactic, and it was determined that hybrids with high yield potential and moderate disease resistance should be selected for mid-Atlantic sorghum production in order to maximize grain yield and quality while minimizing the need for fungicide inputs. Objective 3 focused on integrated management and demonstrated that under moderate disease pressure, a high-yielding susceptible hybrid required a single application of pyraclostrobin-based fungicide to minimize fungal diseases and maintain acceptable yields, whereas under high disease pressure it was necessary to integrate hybrid resistance and judicous applications of fungicides. The aim of Objective 4 was to characterize potential causal agents of mycotoxin contamination in mid-Atlantic sorghum, and thirteen phylogenetically distinct Fusarium species (F. lacertarum, F. graminearum. F. armeniacum, F. proliferatum, F. fujikuroi, F. verticillioides, F. thapsinum and several in Fusarium incarnatum-equiseti species complex) were found to be associated with grain mold and fumonisin and/or deoxynivalenol contamination of sorghum grain. This work has provided insights into the impacts of fungal diseases on grain sorghum yield and quality in the mid-Atlantic and has aided in development of best management practices for the region. / Doctor of Philosophy / Sorghum is grown in tropics, sub-tropics and semi-arid region worldwide for food, feed, forage and fuel. Sorghum acreage in the mid-Atlantic is increasing due the demand for locally grown grain by poultry and swine industries. During the growing season, warm and humid conditions are common in the southeastern and mid-Atlantic states favoring fungal diseases development that reduce the grain yield and quality. Anthracnose and grain mold, which includes toxic mycotoxin-producing Fusarium species, are the two major constraints in sorghum production in the region. However, management alternatives have not been developed. The main goal of this research was to develop management strategies to protect yield and maximize farm profitability by controlling anthracnose and grain mold of sorghum using chemicals and/or host resistance. The specific objectives were to: (1) determine the effectiveness of fungicides and their application timing for the management of sorghum foliar anthracnose, (2) compare grain sorghum hybrids for their susceptibility to foliar anthracnose, grain mold and mycotoxin contamination under field conditions, (3) assess the value of integrating host resistance and judicious use of fungicides to manage sorghum anthracnose and grain mold, and (4) identify Fusarium spp. associated with grain mold and mycotoxin contamination of sorghum in the mid-Atlantic U.S. Results from this research indicate that a single application of pyraclostrobin-containing fungicides no later than flowering reduces anthrancose, protects yield, and increases farm income. Sorghum hybrids varied in susceptibility to anthrancnose and grain mold, and planting a moderately resistant hybrid and applying a fungicide under high disease risk conditions provided the greatest return on investment. Both fumonisin and deoxynivalenol were frequently detected from sorghum grain, and mycotoxin contamination was associated with 13 different Fusarium species from three distinct species complexes. Based on the results of this work, best management practices for minimizing sorghum disease losses were developed for the mid-Atlantic region.

Malts prepared from sorghum and cowpeas that was germinated at 25°C for 0, 1, 2 or 3 days and dried at 60°C for either 2.5, 5, 7.5 and 10 hours, were evaluated for α-amylase, β-amylase and endo-(1,3)(1,4)-β-D-glucanase activity. Germination had a highly significant effect on enzyme activity (P < 0.01). However, cowpea endo-(1,3)(1,4)-β-D-glucanase was not significantly affected by the combined action of germination and drying. β-Amylase was not detected in any of the samples of this experiment. Further analysis using Duncan’s Multiple Range Test, indicated that there was no significant difference in α-amylase activity in samples dried between 5 to 10 hours. Compared to α-amylase, sorghum and cowpea samples had low endo-(1,3)(1,4)-β-D-glucanase activity. / Master of Science

Sorghum (Sorghum bicolor) is an important cereal crop in sub-Saharan Africa. The parasitic weed Striga hermonthica is a major biotic constraint to sorghum production. A novel technology where planting seeds are coated with herbicide to kill Striga that attach to the roots of the host has been shown to be effective in protecting the cereal crop from Striga damage. However, the host plant must have herbicide tolerance. This technology has not been tested in sorghum because there are no herbicide tolerant sorghum varieties available in Kenya and is, therefore, unavailable for subsistence farmers. One of the ways in which genetic variation can be enhanced and herbicide resistance developed is through chemical mutagenesis with ethyl methane sulfonate (EMS). The objectives of this project, therefore, were to: 1) identify sorghum production constraints through farmer PRA in order to determine breeding priorities.in two Striga endemic districts in western Kenya; 2) develop an EMS mutagenesis protocol for sorghum and to enhance the genetic variability of the crop using chemical mutagenesis; 3) evaluate EMS-derived sorghum mutants for improved agronomic performance; 4) develop acetolactate synthase (ALS) herbicide resistance in sorghum and to characterize the mode of inheritance of the trait; 5) determine the effect of herbicide coating of seed of herbicide tolerant sorghum on Striga infestation. In order to determine breeding priorities and constraints in sorghum production and the likelihood of adoption of herbicide seed coating technology, a survey involving 213 farmers was conducted in two Striga endemic rural districts of Nyanza province in Kenya. Results indicated that local landraces like Ochuti, and Nyakabala were grown by more farmers (> 60%) than the improved varieties like Seredo and Serena (48%). Popularity of the landraces was linked to Striga tolerance, resistance to drought, bird damage and storage pests, yield stability and high satiety value. Major constraints to sorghum production were drought, Striga weed, storage pests, bird damage and poverty among the rural farmers. Important characteristics farmers wanted in new varieties were Striga and drought resistance, earliness, resistance to bird and weevil damage and good taste. Striga infestations in sorghum fields were > 70%. Cultural Striga control options were considered inadequate while inorganic fertilization and chemical control were considered effective but unaffordable. Farmers’ willingness to pay a premium of over 30% for a Striga solution gave indication that herbicide seed coating if effective could be adopted by farmers. As a prerequisite to development of herbicide resistance, a comparative study was carried out to determine optimum conditions for mutagenesis and to induce genetic variation in the sorghum. Two sorghum varieties were mutagenized using varying concentrations (0.1 to 1.5% v/v) of EMS and two exposure times (6h and 12h). In laboratory and greenhouse experiments, severe reduction of sorghum root and shoot lengths indicated effective mutagenesis. The LD50 based on shoot length reduction was 0.35% and 0.4% EMS for 6h for Seredo and Kari/mtama-1, respectively. The highest mutation frequency based on chlorophyll abnormalities was 56% for 0.3% EMS for 6h. In the M2 generation, phenotypic variances for panicle characteristics were increased on treatment with EMS. However, significant effects of exposure time and variety indicated the necessity of genotype optimization for some traits. In order to determine the significance of mutation breeding in sorghum, 78 mutant lines derived from EMS mutagenesis, their wild type progenitor (Seredo) and two local checks (Kari/mtama-1 and Serena) were evaluated for agronomic performance in two locations in Kenya. There were significant (P = 0.05) effects among entries for grain yield, 1000-seed weight and visual scores for height uniformity, head exertion, head architecture and overall desirability. The highest yielding entry-mutant line “SB2M13” had a yield of 160% and 152% relative to the wild type (Seredo) and the best check Kari/mtama-1, respectively. Mutant line “tag27” had the highest 1000-seed weight which was 133% relative to the wild type. Seven mutant lines were rated superior to the wild type for panicle characteristics, head exertion and overall desirability. However, the majority of mutants were inferior to the wild type for most characteristics. Superior mutant lines may be developed into direct mutant varieties after multi-location trials or used as breeding material for sorghum improvement. In order to develop acetolactate synthase (ALS) herbicide resistance in sorghum, over 50,000 seeds of Seredo were mutagenized with 0.3% EMS. Over four million M2 plants were screened using 20g ha-1 of the ALS herbicide, sulfosulfuron. Five mutants (hb46 hb12, hb462, hb56 and hb8) survived the herbicide treatment and were confirmed to be tolerant. Mutant lines displayed differential herbicide tolerance, and the general order of tolerance after spray or seed coat application was hb46 > hb12 > hb462 ~ hb56 > hb8. The LD50 values for herbicide application as a spray, or seed coat, showed mutant lines to be up to 20 and 170 fold, respectively, more resistant than the wild type. Chi square analysis of data from herbicide screening of F2 generation of mutant X wild type crosses indicated no difference from the Mendelian segregation of 1:2:1 indicating the herbicide tolerance was inherited as a single semi-dominant gene. Mutant X mutant crosses did not show allelism indicating that the tolerance in all five mutants could be a result of the same gene mutation. To determine effect of herbicide seed coating on Striga infestation, the five herbicide tolerant mutant lines, hb46, hb12, hb462, hb56 and hb8 and the wild type progenitor Seredo were coated with varying concentrations (0.5-1.5% g ha-1) of sulfosulfuron and planted in a Striga endemic field. There were significant (P=0.05) effects of herbicide concentration on Striga density, Striga flowering and seed set, and sorghum plant stand and biomass. All treatments with herbicide coated on sorghum seeds had lower Striga emergence. Coating sorghum seed with 1g ha-1 sulfosulfuron reduced Striga infestation, Striga flowering and Striga seed set by 47%, 52% and 77%, respectively, and was considered the most effective rate as it did not result in sorghum biomass reduction. Mutants displayed differential herbicide tolerance and Striga resistance. Combining seed coating with high herbicide tolerance and inherent Striga resistance would be most effective for Striga control. Overall, the study showed that EMS mutagenesis is effective in inducing variation in sorghum for several traits including herbicide resistance. The mutants developed in this study will be important for sorghum breeding and for protection of sorghum against the Striga weed. / Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2009.