Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Vermicomposts (VCs) are the solid excreta of earthworms, known to contain plant available nutrients, large amounts of microbial life and diversity, and plant growth regulating hormones. VCs may play an integral role in the nitrogen nutrition of Lupinus angustifolius and function to reduce the reliance of legume crops on chemical fertilizers. Furthermore, the effects of the combination of VC solids and VC teas on legume growth and N nutrition, is unknown. The aim of the present study was to determine the role of varying concentrations of chicken manure VC, with and without the additions of varying VC tea concentrations, on the substrate bacterial functional diversity, plant biomass and N nutrition of the legume, L. angustifolius. In the first experiment the plants were grown in pots under glasshouse conditions and VC was substituted into the quartz sand growth media at rates of 5%, 10% and 100%. Furthermore, rhizobia inoculated and non-inoculated groups were established within the VC treatments. The plants were harvested after 30 days and analysed for tissue nutrient concentrations and biomass production. The VC-containing substrates were assessed for wide-spectrum soil analyses, nematode diversity and microbial diversity via Biolog EcoPlates. In the second experiment the plants were grown under similar conditions for 50 days and amended with 5% VC in the growth media as well as 50ml of aerated VC teas. The teas were brewed for 24 hours in concentrations of 4%, 10% and 20% (v/v), with molasses and kelp extract added as sugar sources. The combined treatments of 5% VC and rhizobia inoculation yielded the greatest biomass response. Furthermore the addition of VC allowed for bacterial nitrogen fixation within non-rhizobia treatments. However, changes in VC concentration had no effect on bacterial guild structure but were found to effect nematode funtional diversity. The additions of VC teas to soil media containing 5% VC had no effect on biomass production but were found to influence bacterial nitrogen fixation. Lower concentration teas increased BNF while the 20% tea reduced this parameter significantly over 50 days. The 20% tea also contained significantly greater bacterial functional diversity than the 5% and 10% teas. The findings of study indicate that the combined treatment of VC solids and teas do not increase the plant biomass of L. angustifolius, but that the additions of 20% teas result in greater microbial diversity in the soil. This in turn may lead to increases in soil fertility. Furthermore, additions of high concentration vermicompost teas (20%) shift the dependence of the plant from atmospheric N sources to soil N sources. / AFRIKAANSE OPSOMMING: Geen opsomming beskikbaar

Thesis (MSc)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: This study investigated the activities of several of the enzymes involved in the alternative route of PEP metabolism via PEPc (EC 4.1.1.31). This reaction circumvents the adenyl ate-controlled PK (EC 2.7.1.40) reaction of the conventional glycolytic network under conditions of P stress. It was hypothesized that the synthesis of pyruvate under Pi stress would induce the PEPc alternative route and that C for pyruvate synthesis would primarily be imported via this route. This was assessed by looking at how total enzyme activities are perturbed under P stress and also by following the route of radioactive labelled 14C02 under sufficient (2 mM) and deficient P (2 JlM) conditions in either roots or nodules. The significance of the pathway under P stress, was further assessed by determining pool sizes of pyruvate that was synthesized from PEPc-derived C. The experiments were conducted under glasshouse conditions, as two separate studies: one to investigate the phenomenon of Pi stress and its consequences for PEPc-derived C metabolism, and the other one to study the enzymes involved. Seeds of Lupinus angustifolius (cv. Wonga) were inoculated with Rhizobium sp. (Lupinus) bacteria and grown in hydroponic culture. Tanks were supplied with either 2 JlM P04 (LP) or 2 mM P04 (control) and air containing 360 ppm CO2. Roots experienced pronounced P stress with a greater decline in Pi, compared to nodules. LP roots synthesized more pyruvate from malate than LP nodules, indicating the engagement of the PEPc route under Pi stress. In this regard, pyruvate is considered as a key metabolite under Pi stress. The role of pyruvate accumulation under Pi stress, was further highlighted by the metabolism of PEP via both the PK and PEPc routes. The enhanced PK activities supported these high pyruvate levels. Under P stress, PEPc activities increased in roots but not in nodules and these changes were not related to the expression of the enzyme. Root and nodular PEPc were not regulated by expression, but possibly by posttranslational control. The novelty of our results for symbiotic roots demonstrates that using metabolically available Pi is indeed a more sensitive indicator ofP stress. These results show that under Pi stress, nodules are able to maintain their Pi and adenylate levels, possibly at the expense of the root. It is suggested that nodules do not experience P stress to the same extent as roots or alternatively function optimally under conditions of low P availability. The increase in concentration of pyruvate synthesized from malate, indeed suggest that under LP conditions there is an increase requirement for pyruvate. It is clear from this data that the operation of bypass route in nodules should be investigated further. Nevertheless, this study provided incentives for understanding the role of C pathways in Ni-fixation, in particular under conditions ofP limitation. / AFRIKAANSE OPSOMMING: Die doel van hierdie studie was om die aktiwiteite van verskeie ensieme van die alternatiewe metaboliese roete via phosphoenolpirovaat karboksilase (pEPc, EC 4.1.1.31) te ondersoek. Dié reaksie omseil die adenilaat-beheerde pirovaatkinase (PK, EC 2.7.1.40) reaksie van die konvensionele glikolitiese weg onder toestande van fosfaat (P) stremming. Dit is gepostuleer dat die sintese van pirovaat onder toestande van Pstremming die alternatiewe roete via PEPc sou induseer en dat die koolstof (C) vir pirovaatsintese gevolglik hoofsaaklik vanaf hierdie roete sou kom. Dit is bepaal deur die veranderinge in die totale ensiemaktiwiteite wat sou plaasvind onder P-stremming te ondersoek. Daar is ook gekyk na die roete' wat radioaktiewe C C4C02) sou volg in wortles en wortelknoppies wat behandel is deur blootsteling aan eerder lae fosfaat (2 1lM) of genoegsame fosfaat (2 mM; kontrole), Die betekenis van die alternatiewe roete is ook ondersoek deur die poel-groottes van pirovaat, soos gesintetiseer via die PEPc reaksie, te bepaal. Twee eksperimente is in 'n glashuis uitgevoer. Eerstens is die verskynsel van Pstremming, asook die invloed daarvan op PEPc-afgeleide C-metabolisme, bepaal. Tweedens is die betrokke ensieme bestudeer. Sade van Lupinus angustifolius (cv. Wonga) is geïnokuleer met Rhizobium sp. (Lupinus) bakterieë en in 'n waterkultuur gekweek. Die houers is voorsien met óf2 IlM P04 (LP) óf 2 mM P04 (HP) en lug wat 360 ppm C02 bevat het. Wortels, anders as wortelknoppies, het 'n betekenisvolle afname in anorganiese P (Pi) ervaar. Onder P-stremming, het lae fosfaat wortels meer pirovaat vanaf malaat gesintetiseer as wortelknoppies, wat 'n definitiewe bydrae vanaf die PEPc roete impliseer. Hiervolgens is pirovaat 'n sleutel metaboliet onder P-stremming. Die belangrikheid van die akkumulering van pirovaat onder P-stremmende toestande is verder beklemtoon deur die toename in metabolisme van PEP via beide die PK- en die PEPcreaksies. Die toename in PK-aktiwiteite is goed gekorreleer met die verhoogde produksie van pirovaat. Onder toestande van P-stremming het die aktiwiteit van PEPc in wortels verhoog, maar nie in wortelknoppies nie. Dit was nie die gevolg van 'n verhoogde uitdrukking van die ensiem nie. Wortel- en wortelknoppie- uitdrukking van PEPc is derhalwe nie gereguleer deur die uitdrukking daarvan nie, maar eerder deur post-tranlasie kontrole. Hierdie resultate vir wortels met wortelknoppies demonstreer dat metaboliese Pi 'n beter maatstaf is om P-stres aan te dui. Hierdie resultate toon dat wortelknoppies beter daartoe instaat is om hul Pi-vlakke en adenilaatvlakke te reguleer, en dit mag ten koste van die gasheerwortel wees. Ons stel voor dat wortelknoppies nie P-stremming tot dieselfde mate ervaar as die gasheerwortel nie en dat dié knoppies optimaal funksioneer by lae Pi vlakke. Die verhoogde konsentrasie van pirovaat, wat vanaf malaat gesintetiseer is, impliseer dat daar 'n groter vereiste is vir dié metaboliet onder toestande van Pstremming. Hierdie studie het die rol van koolstofmetabolismein stikstofbindende organismes, spesifiek onder toestande van fosfaat-tekort, beklemtoon.

Thesis (Ph.D.)--University of Western Australia, 2006.

Thesis (Ph.D.)--University of Western Australia, 2007.

[Truncated abstract] Narrow-leaf lupin (Lupinus angustifolius L.) was first recorded as having been introduced into Germany during the mid-19th century for use as green manuring and as fodder crops. However, it was not until post World-War I that there was any serious attempt to domesticate the species. Since that time several key domestication genes have been incorporated to enable the species to be grown as a crop over a range of climates, harvested as a bulk commodity and, the seed used for both animal and human consumption. However, the recent domestication of this species has seen a rather limited use of wild germplasm largely as a result of the difficulty in retaining these key domestication genes. To make the task of retaining these genes manageable, it was decided to resort to molecular technology. A mapping population of F8 derived recombinant inbred lines (RILs) has previously been established by the Department of Agriculture and Food, Western Australia, from a cross between a domesticated breeding line 83A:476 and a wild type P27255 in narrow-leaf lupin. The parents together with 89 RILs (of a population of 115) were subjected to DNA fingerprinting using microsatelliteanchored fragment length polymorphism (MFLP) to rapidly generate DNA markers for construction of a linkage map. Five hundred and twenty two unique markers of which 21% were co-dominant, were generated and mapped. Phenotypic data for the domestication traits: mollis (soft seeds), leucospermus (white flower and seed colour); Lentus (reduced pod-shattering), iucundis (low alkaloid), Ku (early flowering) and moustache pattern on seed coats; were included. Three to 7 molecular markers were identified within 5 cM of each of these domestication genes. The anthracnose resistance gene Lanr1 was also mapped. Linkage groups were constructed using MapManager version QTXb20, resulting in 21 linkage groups consisting of 8 or more markers. ... Five pairs of QTLs were found to be involved in epistasis, 2 of these having an effect on early vigour and another 3 influencing the time to opening of the first florets. Variation explained for each trait ranged from 28% for seed size, to 88% for days to flowering. We showed that it was possible to use this data to predict genotypes of superior progeny for these traits under Mediterranean conditions. QTL regions were compared on a second published linkage map and regions of conserved synteny with the model legume Medicago truncatula high-lighted. The work presented in this thesis demonstrates the importance of tight linkage between markers and genes of interest. It is especially important when dealing with genetically diverse material as found in the wild. One of the main problems faced by molecular scientists is the phenomenon known as linkage disequilibrium in marker populations caused by either small population size or 4 insufficient opportunity for recombination. This frequently results in the development of markers with little or no application outside of the population in which it was developed. Although the relatively small size of the population used in this study exposes it to such constraints, in this case excellent and valuable results were achieved in developing useful markers to at least 3 of the domestication traits within a relatively short time period of less then 4 years.

This diploma work was assessed effects of different seed treatment (untreated flour from whole seed, flour from the uterus after soaking seeds, flour from the seeds sprouted cotyledon) by three species of legumes (Pisum sativum conv. Sativum L., Vicia faba L., Lupinus angustifolius L.) yield and composition of protein isolate obtained by isoelectric precipitation. It was found that the seeds of change most affects yield precipitated N (mg) and 45%. When determining the value of the yield of protein was affected by significant interactions (treatment and the type of legume seeds). The largest share was, however, precipitate in untreated and germinated lupine seeds, in which values are around 57%. Furthermore, the thesis was to reverse the precipitation of proteins, using which we obtained protein concentrates from different species of legumes. The yields of protein concentrates reach values in the range of 60-80%. Spectra of soluble proteins is clearly visible high concentration of isolated proteins. Variation of sprouted seeds and soaked that occurred during treatment of seeds for the collapse of proteins with higher molecular weight. Furthermore, these grafts are transferred protein extraction at pH 9.0 in the later produced protein isolates. After acid precipitation is observed on a spectrum that is re-soluble protein isolates.

To date, there has been little research into the role of microbial community structure in the functioning of the soil ecosystem and on the links between microbial biomass size, microbial activity and key soil processes that drive nutrient availability. The maintenance of structural and functional diversity of the soil microbial community is essential to ensure the sustainability of agricultural production systems. Soils of the same type with similar fertility that had been under long-term organic and conventional crop management in Canterbury, New Zealand, were selected to investigate relationships between microbial community composition, function and potential environmental impacts. The effects of different fertilisation strategies on soil biology and nitrogen (N) dynamics were investigated under field (farm site comparison), semi-controlled (lysimeter study) and controlled (incubation experiments) conditions by determining soil microbial biomass carbon (C) and N, enzyme activities (dehydrogenase, arginine deaminase, fluorescein diacetate hydrolysis), microbial community structure (denaturing gradient gel electrophoresis following PCR amplification of 16S and 18S rDNA fragments using selected primer sets) and N dynamics (mineralisation and leaching). The farm site comparison revealed distinct differences between the soils in microbial community structure, microbial biomass C (conventional > organic) and arginine deaminase activity (organic > conventional). In the lysimeter study, the soils were subjected to the same crop rotation (barley (Hordeum vulgare L.), maize (Zea mays L.), rape (Brassica napus L. ssp. oleifera (Moench)) plus a lupin green manure (Lupinus angustifolius L.) and two fertiliser regimes (following common organic and conventional practice). Soil biological properties, microbial community structure and mineral N leaching losses were determined over 2½ years. Differences in mineral leaching losses were not significant between treatments (total organic management: 24.2 kg N per ha; conventional management: 28.6 kg N per ha). Crop rotation and plant type had a larger influence on the microbial biomass, activity and community structure than fertilisation. Initial differences between soils decreased over time for most biological soil properties, while they persisted for the enzyme activities (e.g. dehydrogenase activity: 4.0 and 2.9 µg per g and h for organic and conventional management history, respectively). A lack of consistent positive links between enzyme activities and microbial biomass size indicated that similarly sized and structured microbial communities can express varying rates of activity. In two successive incubation experiments, the soils were amended with different rates of a lupin green manure (4 or 8t dry matter per ha), and different forms of N at 100 kg per ha (urea and lupin) and incubated for 3 months. Samples were taken periodically, and in addition to soil biological properties and community structure, gross N mineralisation was determined. The form of N had a strong effect on microbial soil properties. Organic amendment resulted in a 2 to 5-fold increase in microbial biomass and enzyme activities, while microbial community structure was influenced by the addition or lack of C or N substrate. Correlation analyses suggested treatment-related differences in nutrient availability, microbial structural diversity (species richness or evenness) and physiological properties of the microbial community. The findings of this thesis showed that using green manures and crop rotations improved soil biology in both production systems, that no relationships existed between microbial structure, enzyme activities and N mineralisation, and that enzyme activities and microbial community structure are more closely associated with inherent soil and environmental factors, which makes them less useful as early indicators of changes in soil quality.

[Truncated abstract] Obesity is now a major public health problem worldwide. More than half the Australian population is now overweight. This is an important public health concern primarily because of the impact of overweight and obesity on risk of diabetes, hypertension and cardiovascular disease. Many strategies have been proposed to fight the obesity epidemic. One possible strategy involves understanding of the role of dietary components in the control of food intake. In this regard, dietary protein and fibre appear to be the most satiating nutrients. Foods enriched in protein, replacing energy from carbohydrate, or dietary fibre can increase satiety and reduce energy intake in the short-term. Longer-term trials suggest benefits of increasing protein or fibre intake on weight loss and features of the metabolic syndrome. The effects of dietary approaches which increase both protein and fibre at the expense of refined carbohydrate are uncertain. A practical approach to increasing both protein and fibre content of processed foods is to incorporate high protein and fibre ingredients into high carbohydrate foods. Lupin kernel flour is a novel food ingredient derived from the endosperm of lupin. It contains 40 to 45% protein, 25 to 30% fibre, and negligible sugar and starch. Lupin kernel flour can be incorporated into refined carbohydrate rich foods such as bread to increase protein and fibre content at the expense of refined carbohydrate. ... Body weight was measured every 2 weeks throughout the 16 week intervention, and these data were analysed to determine whether there was any between group difference in the rate of change in weight over 16 weeks. Over 16 weeks, lupin bread compared to white bread resulted in a significant increase in protein (13.7 (2.3, 25.0) g/d) and fibre (12.5 (8.8, 16.2) g/d) intakes, and a decrease in carbohydrate intake (-19.9 (-45.2, 5.5) g/d). There was a significant difference between groups in the rate of weight change over the 16 weeks (P=0.05). However, at 16 weeks there was no significant effect on body weight (-0.4 (-1.3, 0.6) kg), fat mass (-0.5 (-1.2, 0.2) kg) or fat free mass (0.2 (-0.5, 0.8) kg). Plasma adiponectin and leptin were not altered. Mean 24 hour systolic blood pressure (-2.4 (-3.4, -1.3) mm Hg) and pulse pressure (-3.1 (-3.9, -2.3) mm Hg) were lower for lupin relative to white bread, but diastolic blood pressure was not significantly different between groups. Apart from a lower HDL cholesterol for lupin relative to white bread (-0.09 (-0.17, -0.01) mmol/L), there were no significant differences in other blood lipids and glucose and insulin concentrations. Interpretation of the results was not influenced after adjustment for potential confounding factors. These studies assessed effects of bread enriched in lupin kernel flour relative to white bread, resulting in a higher protein and fibre intake and lower refined carbohydrate intake. This increased satiety and reduced energy intake acutely, but did not significantly influence body weight over 16 weeks. Systolic blood pressure and pulse pressure were significantly reduced. There were no significant improvements in blood lipids or glucose and insulin concentrations. Therefore, increasing protein and fibre intake at the expense of refined carbohydrate using lupin kernel flour may benefit satiety and blood pressure. Longer-term trials incorporating weight loss may be needed to observe benefits on body weight.

A sustainable approach to restoration of the post-mining landscape involves creating conditions where a viable growth medium can be established with the minimal amount of fertiliser inputs over a period of time. Ideally, this growth medium should comprise a combination of minerals, organic matter, and biological components with have the capacity to retain and release nutrients over time. When combined with a re-vegetation scheme appropriate to the biogeographic area, a self-sustaining ecosystem can be established. Regulatory requirements for mine restoration typically require the use of topsoil, which is often in limited supply. However, overburden is commonly present in vast quantities and has the potential to be used as a restoration substrate with appropriate amendments. There are significant gaps in our understanding of factors relating to the use and amendment of overburden as a growing medium. The main objective of this study was to investigate and quantify the effects of inorganic and organic amendments and weathering on nutrient availability and plant growth in topsoil and overburden material obtained from the OceanaGold Globe Progress mine at Reefton, New Zealand. A series of glasshouse experiments were carried out on topsoil, fresh overburden and mullock (old waste rock) amended with biosolids, green manure, sawdust, lime and mineral fertiliser nutrients (nitrogen (N) and phosphorus (P)). R esults demonstrated that 300 kg N ha⁻¹ was sufficient to overcome N deficiency in topsoil, while 100 kg P ha⁻¹ was required to overcome P deficiency. Addition of biosolids to topsoil and overburden increased plant biomass production, without increasing heavy metal concentrations. Amendment of overburden significantly affected weathering rates by accelerating the formation of secondary minerals. In particular, lupin green manure accelerated the formation of organic iron (Fe) and aluminium (Al) organic complexes, resulting from the decomposition of the green manure and release of Fe from primary minerals. Results revealed that wineberry and red beech required 200 kg N ha⁻¹ for optimum growth. Overall, the findings of this research identified key parameters for effective utilisation of overburden as a restoration substrate. A sustainable restoration method for managing the topsoil and/or any overburden or waste rock material remaining at mine sites can be achieved by careful selection of organic amendments and allowing an initial weathering period.

The aim of the diploma thesis was to assess changes in pattern of legume storage proteins during germination. Four species of legumes were chosen for analyses ? Glycine max L., Lupinus angustifolius L., Pisum sativum L. and Vicia faba L. Seeds for analyses were sampled at the beginning, middle and end of germination. Proteins were extracted from lyophilised and homogenised material. These proteins were analysed by SDS-PAGE electrophoresis. The results proved that during seed germination the seed storage proteins cleave into smaller peptides, which forms new proteins. The intensity of protein bands in pea seeds was decreased in the area at around 48-45 kda and 40-36 kDa and the intensity of the proteins bands was increased at around the protein bands 25-23 kDa and 19-7 kDa. In lupine were not detected the protein bands over 39 kDa and during germination amount of protein bands in areas 15-7 kDa was increased