College of Agriculture, Engineering and Science

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Effects of pruning and fertilizer on growth, phytochemistry and biological activity of Sutherlandia frutescens (L.) R.Br.
(2017) Raselabe, Maanda Benjamin.; Van Staden, Johannes.; Finnie, Jeffrey Franklin.; Abdelgadir, Hafiz Ahmed.; Ndhlala, Ashwell Rungano.
Sutherlandia frutescens (L.) R.Br. (Fabaceae), commonly known as cancer bush, is a herb with a long history of traditional use by a variety of cultures. The plant mainly grows in the dry parts of southern Africa, mostly in the Western and Eastern Cape as well as the neighbouring countries like Lesotho, Botswana and Namibia. Cancer bush is traditionally used for the treatment of external wounds, internally for fevers, stomach problems, cancer, diabetes, influenza, HIV, depression, eye problems, TB, colds and asthma. The plant is famously known for the treatment of Type 2 diabetes, cancer and HIV. However, these claims remain inconclusive. Recent studies have shown S. frutescens to have antidiabetic, anti-HIV, anti-inflammatory, antibacterial, analgesic, anti-stress, anticonvulsant, antiproliferative and antithrombotic activities. Phytochemical investigations of S. frutescens leaves detected the presence of high levels of free amino acids and non-protein amino acids namely: canavanine, gamma aminobutyric acid (GABA) and pinitol. The presence of these compounds has been reported to be responsible for its reputed effectiveness in a wide range of illnesses. In view of its importance as a multipurpose medicinal crop, it is important to bring this plant under cultivation and determine agronomic requirements for its successful cultivation. Several factors can be investigated to enhance the growth and increase the level of active ingredients. The current study was aimed at evaluating the effect of pruning and fertilizer levels on the growth, phytochemistry and biological activity of Sutherlandia frutescens. Seeds were sown in seedling trays to produce seedlings. One-month-old seedlings were then transplanted on a prepared field. The study trial was carried out at the Agricultural Research Council-Vegetable and Ornamental Plants Research Station (ARC-VOP). The experiment was conducted in a randomized complete block design (RCBD) with three pruning levels, three fertilizer levels and four replicates. There were 9 treatments, namely; no pruning (P0), tip-pruning (P1) and heading back (P2) in combination with levels of fertilizers as follows: 200 kg/ha NPK (F1), 100 kg/ha NPK (F2) and 0 kg/ha NPK (F0). Plants without treatments were considered as controls. Growth parameters taken were plant height, stem diameter, chlorophyll and Leaf Area Index (LAI). Dried leaf samples were analysed for the presence of secondary metabolites and antidiabetic activities. There was no significant interaction effect between pruning and fertilizer levels amongst all parameters measured in this study. Pruning treatments had a significant effect on the LAI at week one and week two but did not affect the plant height, stem diameter and chlorophyll content. Different levels of fertilizers had a significant effect on the LAI, where 100 kg/ha NPK significantly increased LAI at week one and week two. In an investigation which was conducted from January to May 2015, plants showed yellowing, stunting, and high levels of infestation expressed as extensive galling on the roots which led to the nematode infestation study. Nematodes were extracted from the roots of a healthy living, a wilted and a dead plant, as well as from the rhizosphere soil. A small population of Scutellonema, Pratylenchus, Helicotylenchus and Tylenchorhynchus were identified. Examination of the root of an infected plant revealed the presence of root-knot nematodes (Meloidogyne javanica) in large numbers. Juveniles, eggs and females were isolated, and the species were identified on the basis of morphological characteristics. Symptoms usually associated with root-knot nematodes were observed on the roots of the wilted plant and the soil in which the plant was growing. Extracts from all nine treatments showed stronger activity against α-glucosidase than the positive control acarbose. The highest α-glucosidase inhibitory activity was demonstrated by the treatment with no pruning (P0) while the heading back (P2) treatment exhibited the lowest inhibitory activity. Fertilizer levels at 200 kg/ha (F1) NPK resulted in a significantly higher α-glucosidase inhibitory activity compared to other fertilizer treatments. The presence of secondary metabolites (including total phenolics and flavonoids) was determined qualitatively. The total phenolic content was determined using the Folin-Ciocalteu method and flavonoids were determined using the vanillin HCL assay. The study showed that pruning and fertilizer increased the production of secondary metabolites in S. frutescens as compared to the control. Fertilizer at 200 kg/ha NPK (F1) did seem to improve phenolics and flavonoids with pruning but phenolic levels were actually quite low when treatment was P0F1. Total phenolics and flavonoids were significantly increased by the heading back (P2) treatment and decreased in the treatment with no pruning (P0). The application of fertilizer at 200 kg/ha NPK (F1) improved the production of secondary metabolites, and reduced production of secondary metabolites was recorded in plants that received no fertilizer. There was no direct correlation between the level of phytochemicals and the antidiabetic activity recorded. This study examined the effects of different levels of pruning and fertilizers on the growth of S. frutescens. The results showed that there was no significant difference. At this stage, no positive recommendations can be made for cultivating S. frutescens. Plant extracts showed good antidiabetic activities in response to different pruning and fertilizer treatments. This was further seen as an increase in the production for secondary metabolites. However, further investigation of plant cultivation practices and further screening for bioactivities is required. S. frutescens may offer a new source of drugs for diabetes mellitus and other related diseases.
The potential of Moringa oleifera (Lam.) leaves for use in complementary foods to combat child food and nutrition insecurity among South African rural and peri-urban communities.
(2017) Ntila, Sithandiwe Linda.; Siwela, Muthulisi.; Kolanisi, Unathi.; Abdelgadir, Hafiz Ahmed.; Ndhlala, Ashwell Rungano.
The inadequate nutritional quality of complementary foods is a major public health problem in developing countries such as South Africa, and contributes towards the reported high rates of child malnutrition. Consequently, there is an urgent need to investigate interventions aimed at improving the nutritional quality of complementary foods. The aim of this study was to assess the potential of Moringa oleifera (Lam.) leaves for use in complementary foods to combat food and nutrition insecurity amongst children in South African communities. Mothers from Lebowakgomo village in Limpopo province (n=106) and Hammanskraal Township in Gauteng province (n=106), were recruited on a voluntary basis to participate in the survey which assessed the food and nutrition security status of their children, aged 7-12 months old. Additionally, six focus group discussions were conducted per study area to assess mothers’ perceptions about children’s food access and to further establish a recipe for a popular traditional cereal-based complementary food. The Children’s Food Insecurity Access Scale (CFIAS) was used to assess the food security status (access) of the children. The Individual Dietary Diversity Score (IDDS) together with the unquantified food consumption frequency survey were used as a proxy measure of the nutritional quality of the children’s diets. The age and weight of the children were obtained from the children’s clinic health cards and were used to calculate Weight-for-Age Z scores (WAZ). These values were interpreted to determine the prevalence of underweight children. The popular cereal-based traditional complementary food recipe (white maize soft porridge) used by mothers from the studied communities was modified by substituting maize meal with MLP at 1, 2 and 3% (w/w) levels. The nutritional, phytochemical and antioxidant analysis of the popular traditional complementary soft porridge (control) and Moringa-added porridges (test samples) were determined using standard methods. The sensory acceptability of the porridges was evaluated by the mothers who were recruited on a voluntary basis (n= 60 per study area) and six focus group discussions were conducted per study area to assess mothers’ perceptions on the inclusion of Moringa in complementary foods. The findings of the survey showed that a large percentage of children were severely food-insecure, 87% and 78%, at Lebowakgomo and Hammanskraal, respectively. Additionally, children from Lebowakgomo (23.6%) and Hammanskraal (17.9%) were severely underweight. Overall, children’s diets in both study areas were characterized by nutrient-deficient complementary foods. Cheaper foods with a higher satiety value such as white maize meal (WMM) and sugar were the most commonly purchased and used. Hence, the children consumed very limited amounts of foods rich in proteins, minerals, and vitamins, which significantly increased the risk of malnutrition. As the MLP was increased from 0 to 3% in the Lebowakgomo white maize soft porridge, a corresponding significant increase in nutrient content was observed: ash (from 0.52 to 0.87 g/100 g), calcium (0.01 to 0.09 mg/100 g), potassium (0.10 to 0.14 mg/100 g), protein (8.70 to 9.68 g/100 g), threonine (0.14 to 0.66 g/100 g), glutamine (1.28 to 1.56 g/100 g), provitamin A (0.81-1.16 μg/g), lutein (0.04-0.30 μg/g), zeaxanthin (0.21-2.18 μg/g), β-cryptoxanthin (0.11-0.14 μg/g), β-carotene (0.25-0.50 μg/g) and 9-cis-β-cryptoxanthin (0.25-0.31 μg/g). Increasing the MLP from 0 to 3% in the soft porridge at Hammanskraal, caused a significant increase in the levels of iron (from 52.50 to 101.0 mg/100 g), manganese (1.00 to 4.00 mg/100 g), phenylalanine (0.35 to 0.47 g/100 g), provitamin A (0.87-1.01 μg/g), lutein (0.05-0.22 μg/g), zeaxanthin (0.22-1.29 μg/g) and β-carotene (0.27-0.39 μg/g). Additionally, the antioxidant activity, total phenolic and flavonoid contents of Hammanskraal and Lebowakgomo white maize soft porridges increased as the concentration of MLP was increased. The sensory evaluation results showed that the acceptance of each of the two traditional complementary foods decreased as the level of MLP increased. Mother’s indicated in the focus group discussions that Moringa-added soft porridges had a bitter taste, which would not be suitable for children. Only the Moringa-added soft porridge from Lebowakgomo containing 1% of Moringa was rated similar in overall acceptability as the corresponding white maize soft porridge (control). Nevertheless, all mothers expressed willingness to use Moringa in complementary foods if they would be trained on how to process it into foods suitable for children. There is a need to vary product formulation and processing methods, which may contribute to increased acceptance of Moringa-based foods. Additionally, the safety of using MLP in complementary foods should be investigated.
Strategies to improve seed production in Jatropha curcas : a potential seed oil crop for biodiesel.
(2009) Abdelgadir, Hafiz Ahmed.; Van Staden, Johannes.; Johnson, Steven Dene.
Interest in planting Jatropha curcas L. for the production of biodiesel is growing exponentially. The properties of the crop and its oil have persuaded investors to consider J. curcas oil as a substitute for fossil fuels. However, this plant is still undomesticated, basic agronomic properties are not thoroughly understood and the environmental effects on growth have not been investigated. This thesis investigated different approaches that may contribute to improving the productivity of this plant. Seed germination and methods of propagation are usually the first consideration in any plant development programme. The effects of aerosol smoke, smoke water, potassium nitrate, naphthalene acetic acid and indole-3-butyric acid on germination and seedling growth of J. curcas were investigated. Seed coat removal accelerated water imbibition and germination occurred within 48 h. Seeds exposed to aerosol smoke failed to germinate over the whole study period of three months. There were no significant differences in total germination between the treatments and the untreated control (intactand shelled-seed). However, shelled-seeds had a shorter mean germination time. The seedlings were subsequently sown in trays under shade house conditions and different seedling growth traits measured after three months. Smoke water, potassium nitrate and naphthalene acetic acid produced significantly heavier seedlings with longer stems and roots, wider stems and a higher vigour index compared to the control treatments. Smoke water, potassium nitrate and naphthalene acetic acid stimulated seedling growth and vigour of J. curcas. This opens the possibility of applying these treatments to produce quality seedlings for large scale planting and accelerated plant establishment in production orchards. Effective pollination is a prerequisite for many crops to increase seed-set and fruit production. Experiments were conducted to determine factors that could influence seed production in this potential biofuel seed crop. Controlled pollination experiments showed that plants required pollinator visits for seed production and were genetically selfx compatible. Pollen-supplementation did not lead to increased fruit set, suggesting that seed production in the study population was not pollen-limited. Both male and female flowers produced nectar and were highly attractive to honeybees. These insects were effective pollinators of J. curcas, as shown by experiments in which flowers exposed to single or multiple visits by honeybees set significantly more fruit than those from which visits were precluded. Pollinator-mediated self-pollination led to marginally lower levels of seed production relative to cross-pollination. Progeny from selfed plants had significantly shorter roots than progeny of outcrossed plants. However, in general, there was little evidence of inbreeding depression. The present results provide empirical evidence that honeybees are effective pollinators of J. curcas. Fruit arising from selfpollination were almost as numerous and as large as those arising from crosspollination, suggesting that promotion of cross-pollination does not have to be a priority in orchard management for fruit yield. Manipulation of pollen development and function is of vital importance for crop development and improvement. Experiments were conducted to investigate pollen viability, in vitro pollen germination and in vivo pollen tube growth in J. curcas. Light and fluorescence microscopy were employed to examine the different developmental stages. It was possible to determine pollen viability and distinguish between fresh and dead pollen using 2,3,5-triphenyltetrazolium chloride (TTC). Pollen germination was significantly higher in an agar-based medium composed of sucrose, boric acid and calcium nitrate compared with the control treatment (distilled water). Supplementation of IAA to the different media significantly increased pollen germination and pollen length compared with the control treatment. Pollen from hermaphrodite flowers had a lower viability, lower germination rates and shorter pollen tubes, with abnormal shapes, compared to the pollen from male flowers. Pollen tubes from both self- and crosspollinated flowers entered the ovary within 8 hours after pollination (HAP). However, at 6 HAP, the pollen tube length and growth rate were significantly higher in crosscompared to self-pollinated pollen. Our results suggest that TTC is a reliable test for pollen viability; boric acid, calcium nitrate, sucrose and addition of IAA are essential and beneficial for pollen germination in this plant. Pollen germination and pollen tube growth were not inhibited, nor interfered with, as a result of self-pollination treatments. During, both types of pollination, fertility is maintained as evidenced by ovule penetration by pollen tubes. This suggests that type of pollination has no influence on the success of fertilization in J. curcas. Manual pruning is one of the major management practices in commercial plantations of J. curcas, resulting in production of more branches and thus increased potential for more inflorescences leading to a higher seed yield. Experiments were conducted to determine the response of J. curcas plants to manual pruning under summer and winter conditions. The results showed that manual pruning under both conditions significantly increased the number of branches per plant. However, there were no significant differences in number of branches between winter and summer manual pruning. Winter pruning, however, had a significantly wider crown diameter compared to the control and summer pruning. Both treatments produced significantly less fruits/per plant in the subsequent season compared to the un-pruned control. This study revealed that winter and summer manual pruning may be suitable practice to promote branching. Manual pruning, however, is time consuming, labour intensive and expensive. A study was conducted to determine the potential of different plant growth regulators (PGRs) to increase the number of lateral branches of J. curcas plants. A single foliar application of BA (benzyladenine) at 12 mmol l-1 significantly increased branches in both the pot (4) and field (13.2) trials compared to manual pruning (MP) (1.8 and 5.7 respectively) and control (no new branches) plants. In the field, treatment with TIBA (2,3,5-triiodobenzoic acid) (1 mmol l-1) significantly increased the number of branches (15.9) after seven months from application. Of all the PGRs examined, DK (Dikegulac) (2,3:4,6-di-Oisopropylidene- 2-keto-L-gulonic acid) at 2 mmol l-1 produced the maximum number of branches (18) in the field seven months after application. Concentrations of 2 and 3 mmol I-1 of MH (Maleic hydrazide) (1,2-dihydro-3,6-pyridazinedione, coline salt) significantly increased the number of branches, four and seven months after spraying in both the pot trial in the shade house and field respectively. Under field conditions J. curcas plants responded better to all the PGRs (DK < TIBA < BA < MH) when treated once, with insignificant variations of other growth parameters. This study indicates that a single foliar application of PGRs under field conditions can be an alternative method to MP for increasing the number of lateral branches of J. curcas plants. The field chemical pruning experiment was continued to determine the potential subsequent effects of the different PGRs on seed production. In the subsequent year following the single foliar application, the parameters of flowering, fruit set, fruit characteristics, total oil content and free fatty acid (FFA) content were evaluated. Number of flowers per plant and number of fruits per bunch were significantly affected by the different treatments. However, there were no variations in the degree of fruit set. A single foliar application of BA (6-benzylaminopurine) produced more flowers per plant, more fruits per bunch, heavier and bigger fruits and seeds with more oil compared to MP (manual pruning). TIBA (2,3,5-Triiodobenzoic acid) produced significantly more flowers per plant and heavier fruits compared to the control and MP treatments. However, it produced significantly bigger fruits with more seeds and a higher oil content than MP. DK (Dikegulac) (2,3:4,6-di-O-isopropylidene-2-keto-Lgulonic acid) produced more flowers per plant and seeds with high oil content compared to the control and MP. However, it produced more fruit per bunch and more seeds per fruit compared to MP. MH (Maleic hydrazide) produced more flowers per plant, heavier and bigger fruits with numerous, heavier and oil rich seeds compared to the control and MP. This study indicates that foliar application of PGRs can be used in J. curcas to increase seed production and improve fruit quality.

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