>Magister Scientiae - MSc / Molecular methods were used to investigate the microbial diversity and community structure of ammonia-oxidizing bacteria (AOB) associated with the roots of the Proteaceae plant family. The identification of ammonia oxidizing bacteria in this ecosystem is of particular interest since Proteaceae are adapted to acidic, low nutrient (e.g. nitrogen) soils. The ammonia monooxygenase operon was used as a molecular marker to identify ammonia-oxidizing bacteria associated with the proteoid roots of the three Proteaceae members and compared to non-plant associated soil. PCR amplification using primer sets targeting the ammonia monooxygenase gene (amoA subunits) were used to construct a clone library. Sequence diversity was determined by RFLP analysis of amoA to identify major groups of AOB of the ~-subclass of Proteobacteria in total community DNA, and DNA sequencing and phylogenetic analysis were also applied. DGGE analysis was performed to determine the community structure and distribution of ammonia-oxidizing bacteria in plant-associated and non-plant associated soils. The AOB genotypic diversity was similar in the plant-associated samples and non-plant associated soil. All AOB phylotypes belonged to Nitrosospira species and clustered with Nitrosospira cluster 3. The abundance of the amoA was quantified to be approximately 4.2 x 107 copies/g of dry soil, using a real-time PCR assay. These data suggest that the Nitrosospira species are the dominant phylotypes in that environment. This investigation provides new insights into the relationships between plants and ammonia-oxidizing bacteria in natural Fynbos ecosystems.

Thesis (MSc)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: The commercial cultivation of Proteaceae is an important industry in the Western Cape, however, farmers are challenged with arthropod infestation which compels them to solely rely on chemical pesticides. Past studies in South Africa have shown that Proteaceae comprise a rich and diverse arthropod fauna. However, as most of these studies were conducted on wild Proteaceae, they may not be representative of cultivated proteas. Moreover, most of these species remained unidentified due to lack of identification expertise. These past studies, however, form a useful baseline for arthropod studies in proteas, e.g. the feeding guilds found in proteas. The aim of this research was to conduct an intensive and extensive survey of the arthropod-fauna associated with commercially-cultivated proteas across an entire year. Specifically, this survey was designed to document the composition of the arthropod fauna (creating a comprehensive reference collection for pest management purposes) and to assess whether the arthropod fauna differed between seasons and pesticide treatments. Infructescences, inflorescences and foliage of mainly commercial Proteaceae were sampled for arthropods seasonally for a period of twelve months by collection of plant material and direct searching. Seven commercial protea blocks, and a wild protea block (remnant patch of fynbos vegetation), were used as the sampling sites, and two sprayed blocks were used for assessing pesticide efficacy. Individual arthropods were identified as far as possible, with 37% identified to species level. A species accumulation curve showed that rare (minor) arthropod species made up of 70% of arthropods occurring in cultivated proteas. More than 8 700 individuals from more than 140 species and about 80 families were collected and identified, revealing that cultivated proteas have a rich and diverse insect fauna. These arthropods represent the full range of plant-feeding guilds: leaf miners, leaf chewers, flower bud borers, sap suckers and seed feeders. Flower visitors/free living guild was the most abundant (72%) and speciose (25%). In addition to phytophages, there was a large suite of insect predators and parasitoids. A large number of the arthropods were endemic to the Cape Floristic Region (CFR) and some (7.86%) have a pest status, in that they cause significant damage to the protea plants (for example, 60% of Safari sunset cultivar (Leucadendron salignum x L. laureolum) new flush stems and leaves were affected by Epichoristodes acerbella (Tortricidae). Capys alphaeus (Lycaenidae) and Phyllocnistis sp. (Phyllocnistidae) appear to be specialist pests, as they attack mainly Protea cynaroides and Susara cultivar (Protea magnifica x P. susannae) respectively. Arthropod abundance did not differ significantly between seasons, although significant seasonal effects were observed in species richness when the protea cultivars were examined separately. Pesticide application did not affect arthropod abundance, but did decrease species richness in sprayed blocks. Pesticides appeared to negatively affect minor (rare) species disproportionately, probably due to their lack of prior exposure to pesticides and hence sensitivity. Due to this inefficacy of pesticides in cultivated proteas, an increasing emphasis on the importance of non-chemical control measures, and our improved knowledge of the predatory and parasitic species in this system, integrated pest management strategies deserve greater research attention. Monitoring and use of threshold values for arthropod pests were suggested here, as well as the use of biological, cultural, physical and chemical (optimal use) control. For instance, in cultural control, polycropping and intercropping in proteas to increase plant diversity in the monocultures to promote a higher density of predators and parasitoids can be used. Certain flowering plants are known to provide greater temporal and spatial distribution of nectar and pollen sources, which can increase parasitoid reproductive potential and abundance of alternative hosts/prey when the pest species are scarce or at an inappropriate stage. / AFRIKAANSE OPSOMMING: Die kommersiële verbouing van Proteaceae (proteas) is 'n belangrike bedryf in die Wes-Kaap. Menige plantasie wemel egter van artropodes, wat boere noop om slegs van chemiese plaagdoders gebruik te maak. Vorige studies in Suid-Afrika toon dat proteas die gasheerplant vir 'n ryke en diverse artropodefauna is. Aangesien die meeste van hierdie studies egter op wilde proteas uitgevoer is, weerspieël dit moontlik nie die stand van sake met verboude proteas nie. Weens 'n gebrek aan kundigheid om die artropodes te eien word baie van die spesies boonop nooit uitgeken nie. Dié studies voorsien egter 'n nuttige grondlyn vir 'n ondersoek na die artropodes op proteas, veral vir die bestudering van die gilde wat van die protea leef (“the feeding guild”). Hierdie navorsing het ten doel om 'n intensiewe en omvattende opname te maak van die artropodefauna wat oor die tydperk van 'n jaar op kommersieel verboude proteas voorkom. Die opname is meer bepaald ontwerp om die samestelling van die artropodefauna te bestudeer (deur 'n omvattende verwysingsversameling vir plaagbestuurdoeleindes te skep), en om vas te stel of seisoene en plaagbehandelings enige beduidende uitwerking op die artropodefauna het. Oor 'n tydperk van 12 maande is seisoenale monsters van die vrug- en bloeistadia, saadkoppe en blare van hoofsaaklik kommersiële proteas gesoek en ingesamel. Sewe kommersiële proteablokke sowel as 'n blok wilde proteas het as proefpersele gedien, en twee bespuite blokke is gebruik om die doeltreffendheid van plaagdoder te beoordeel. Individuele artropodes is so noukeurig moontlik uitgeken – 37% tot op spesievlak. Volgens 'n spesieakkumulasiekurwe maak seldsame (kleiner) artropodespesies sowat 70% van die artropodes uit wat op verboude proteas voorkom. Die meer as 8 700 individue van meer as 140 spesies en sowat 80 families wat ingesamel en uitgeken is, toon die rykheid en diversiteit van die artropodefauna op verboude proteas. Hierdie artropodes verteenwoordig die volle reeks plantvreterspesies – van blaardelwers en blaarkouers tot blomknopboorders, sapsuiers en saadvreters. Blombesoeker-/vrylewende spesies was die volopste (72%) en mees divers (25%). Buiten plantvreters was daar ook 'n groot aantal roofinsekte en parasitoïede. Baie van die artropodes was inheems, en sommige (7,86%) het boonop plaagstatus, aangesien hulle beduidende skade aan die proteaplant aanrig. [By ongeveer 60% van die Safari Sunset-kultivar (Leucadendron salignum x L. laureolum) is nuwe stamme en blare byvoorbeeld deur die Epichoristodes acerbella (Tortricidae) aangetas.] Capys alphaeus (Lycaenidae) en Phyllocnistis sp. (Phyllocnistidae) blyk spesialisplae te wees wat onderskeidelik hoofsaaklik die Protea cynaroides en die Susarakultivar (Protea magnifica x P. susannae) in die visier het. Artropodegetalle het nie juis tussen seisoene gewissel nie, hoewel 'n afsonderlike ondersoek van die proteakultivars 'n beduidende seisoenale uitwerking op spesierykheid aan die lig gebring het. Eweneens het die toediening van plaagdoder nie die artropodegetalle verminder nie, maar wel spesierykheid op die bespuite blokke verswak. Plaagdoders blyk besonder negatiewe uitwerking op kleiner (seldsame) spesies te hê – waarskynlik omdat dié spesies nie voorheen aan plaagdoders blootgestel was nie, en dus gevoelig is daarvoor. Weens die oënskynlike ondoeltreffendheid van plaagdoders op verboude proteas, verg 'n toenemende klem op die belang van niechemiese beheermaatreëls, 'n behoefte aan meer kennis van die roof- en parasitiese spesies in die stelsel, en die vraag na geïntegreerde plaagbeheerstrategieë, meer navorsing. Die studie moniteer en gebruik drempelwaardes vir artropodeplae, sowel as biologiese, kulturele, fisiese én chemiese (‘optimalegebruik’-) plaagbeheer. Met kulturele beheer kan poli- en interverbouing van proteas byvoorbeeld gebruik word om plantdiversiteit in die monokulture te verbeter, ten einde só 'n hoër digtheid van roofspesies en parasitoïede in die hand te werk. Sekere blomplante bied kenmerkend 'n wyer tyd- en ruimtelike verspreiding van nektar- en stuifmeelbronne, wat parasitoïede se voortplantingsvermoë en die getalle van alternatiewe gashere/prooi kan verbeter wanneer die plaagspesies skaars is of in 'n ontoepaslike stadium verkeer.

[Truncated abstract] In south-western Australia a rare plant community occurs on shallow soils overlaying massive ironstone rock. These 'ironstone communities' are open shrublands, which are subject to extremes in drought and solar radiation and support many rare and endemic species. The restricted distribution of many of these species may be related to their high degree of specialisation to this harsh habitat and their inability to respond plastically to different environmental conditions. Indeed, earlier work has shown that ironstone Hakea species (Proteaceae) have a specialist root-system morphology investing mainly in deep roots, thereby increasing their chance of accessing cracks in the rock surface and obtaining water before the onset of summer drought. In this thesis I further examine aspects of specialisation and its possible consequences for species rarity using two ironstone Hakea species and comparing them with two of their widely distributed congeners. In the first experiment (Chapter 2) I explore inherent drought tolerance, independent of root-system morphology, as a further specialisation to the ironstone environment. All species were grown in sand in pots in a glasshouse for 7 months and then droughted for 5 weeks. There was no evidence that the ironstone species had a greater inherent drought tolerance than their common congeners. During drought all species maintained leaf water content of mature leaves by reducing stomatal conductance and osmotically adjusting, though ironstone species tended to OA (osmotic adjustment) more than common species. ... This suboptimal investment of resources may result in a lower competitive ability in shadier environments, and thus could partially explain their restricted distribution. In Chapter 4, I investigated the plasticity of root traits in response to levels of phosphorus supply. South-western Australian soils are phosphorus impoverished and phosphorus is well known to elicit plastic responses in root allocation and architecture. Ironstone species showed less plasticity in total root length, producing similar root length across P treatments, while common species showed an increase in root length with increasing [P]. Other root characteristics were similarly plastic in response to P treatment between species. However, when supplied with increasing [P], ironstone species invested an increasing proportion of roots in the bottom of pots while common species invested more in the top. This differential response in root allocation in response to P may reflect a fundamental trade-off between nutrient and water acquisition, with the ironstone species mainly foraging for water and investing in deeper roots, while the common species invest more in superficial roots to obtain nutrients. In conclusion, the rarity and restricted distribution of the ironstone Hakea species may be related to their specialist root-system morphology as well as a lowered phenotypic plasticity of functional traits. A reduction in plasticity may reduce their competitive ability outside their ironstone habitats, and thus contribute to the restricted distribution of these species. This may also be the case for other rock-outcrop endemics and more generally, for other rare plant species restricted to particular habitats where a lowered phenotypic plasticity in traits relevant to their particular habitat may contribute to their restricted distribution.

A range of Proteaceous shrubs and other nursery plants were grown in containers with soilless media and various N levels. Plants demonstrated a range of responsiveness. Supplying soilless media with Osmocote (26% N) and other short term fertilisers proved to be a satisfactory method of studying the comparative nutrition of a wide range of container grown nursery plants using factorial experiments incorporating N, P, K and lime. Nutrient response surfaces were obtained using a central composite incomplete block design. Most Proteaceous shrubs were intolerant of high P levels due to excessive luxury consumption resulting in toxic foliar nutrient levels, especially in the presence of high N. Phosphorus sensitivity in plants appeared to correspond with the soil nutrient levels in their native habitat. This applied to species studied in the 2 main sub-families in the Proteaceae while similar findings were indicated for other Australian genera. A range of optimum N requirements in the Proteaceae was found – lowest needs for Protea which also had the greatest tolerance of very low fertiliser additions, compared to Grevillea robusta with much higher N requirements and strong foliage growth inhibition if nutrient levels are very low. Proteoid root growth on Grevillea rosmarinifolia only occurred at low nutrient levels and was not required for satisfactory foliage growth of container grown plants. Pot plants and seedlings, especially tomato, responded strongly to N and often there were positive NK interactions influencing foliage growth. Lime requirements were studied and in erica increasing lime rates depressed foliage growth. Comparative nutrition studies indicate that general or broad spectrum container media may be unsuitable for some groups of nursery plants and that they could be replaced by potting mixes designed to meet the widely differing needs of the species often grown. The number of specialist mixes would depend on the range of plants and be governed by management considerations.

[Truncated abstract] The species-rich Southwest Australian Floristic Region (SAFR) is a global biodiversity hotspot. Characterised by a Mediterranean-type climate and nutrient deficient landscape, this region is endowed with 7380 native vascular plant species/sub species, of which 49% are endemic and 2500 are of conservation concern. Despite the global significance of this region, there is still only a poor understanding of the factors influencing high diversity and endemism, and especially the population genetic consequences of narrow endemism and naturally fragmented species distribution. Holly leaved banksia (Banksia ilicifolia R. Br.), although widespread through Southwest Western Australia (SWWA), has a naturally fragmented distribution, with generally small populations restricted to swales and wetland fringes with depth to groundwater less than 10 m. As such, it provides an excellent model to better understand the ecological genetic consequences of local endemism, population size and natural population fragmentation . . . Products of wide outcrossing (over 30 km) showed a heterosis effect over local outcrossing, indicating increased ecological amplitude of offspring following interpopulation mating. These results suggest that the breeding and mating biology of B. ilicifolia counters the negative genetic erosion effects of narrow ecological amplitude and small population size. Recent habitat fragmentation, and reductions in population size and increased isolation, is impacting on these processes, but further research is required to assess the ultimate consequences of these genetic effects for population viability.

Thesis (DTech (Tourism and Hospitality Management))--Cape Peninsula University of Technology, 2009 / The growing global perspective on conservation in combination with the rapid changes in the environment due to ever increasing human demands has placed more emphasis on the plight of threatened and endangered Proteaceae on the Agulhas Plain. Large parts of the Agulhas Plain contribute to the commercial cut flower export industry. The population numbers of Leucadendron elimense subsp. elimense, L. laxum, L. platyspermum and L. stelligerum (PROTEACEAE) have been drastically reduced as many species are illegally harvested. The continued destruction of natural habitats has made environmentalists and Protea flower producers aware of the need for developing future conservation strategies. An increase in ecotourism on the Agulhas Plain is important in view of its role in extended conservation and job creation. The Agulhas Plain is a region where conservation, ecotourism and agriculture could work together to maintain a balance of protection, enjoyment and commercial gain from the habitat. Potential developments of ecotourism on the Agulhas Plain are hindered through poor agricultural practices and a lack of conservation of the natural habitat. Landowners and cut flower producers have to stay abreast of global changes if they are to be responsible for the protection of the environment. In this respect the usage of land is linked to skills, attitudes, knowledge and an understanding of the environment. There is a lack of guidance and available information in the conservation of the Agulhas Plain while the ecotourism potential of the Cape Floral Kingdom remains undeveloped. The objectives of the study are: a) to collect and survey scientific data on current practices of landowners, flower producers and exporters to determine the probable causes of destruction of Red Data species and their influences on ecotourism development on the Agulhas Plain. This study aims to make recommendations on the propagation and the conservation of threatened species and the ecotourism potential on the Agulhas Plain; b) to test the rooting ability of L. laxum using four liquid hormone concentrations of IBA or IAA and four different rooting mediums. Differences in rooting in an environmentally controlled greenhouse environment with bottom heat and a shaded tunnel is also tested. The study aims to develop new propagation techniques to increase successful and economical propagation of the species, to solve problematic and difficult propagation techniques and to relieve the threatened status of species. The study used a self-administered survey questionnaire distributed amongst growers, farmers and exporters to determine the knowledge, skills, values and attitudes in the harvesting, propagation, conservation and ecotourism development of Red Data species on the Agulhas Plain.

Thesis (MSc)--Stellenbosch University, 2011. / ENGLISH ABSTRACT: With growing concerns about current environmental issues, such as climate change, that affect ecosystems around the world, understanding ecosystem function is becoming increasingly important. In this study, I investigate the plant – seed disperser mutualism between an endemic scatter-hoarding mouse Acomys subspinosus and its Proteaceae plant counterpart Leucadendron sessile in the biodiversity hotspot of the Cape Floristic Region, South Africa. The main objective of this thesis is to investigate the seed selection and caching behaviour of A. subspinosus. First, I determined the seed selection strategy for dispersal and burial by A. subspinosus. Acomys subspinosus may exert stabilizing selection pressure onto L. sessile seeds by dispersing and burying medium seeds with medium hull-thicknesses. Small seeds were eaten in situ and large seeds left at depots. I concluded that the buried L. sessile seeds may have a competitive advantage when it comes to seedling establishment in a post-fire environment, since seeds dispersed by rodents in the fynbos, such as L. sessile, are much larger in size and therefore have more stored nutrients and rapid growth capabilities than seeds dispersed by other vectors. Secondly, I documented rodent dispersal behaviour over a full years’ time. Acomys subspinosus dispersal behaviour changed significantly over the seasons, which I attributed to a change in food availability as the year progressed. Acomys subspinosus buried seeds in autumn after mass seed drop but began to recover caches and consume seeds during winter and spring. The rodent switched to an insectivorous diet in spring. I propose that the A. subspinosus – L. sessile relationship is mutualistic during the year, but the relationship does shift in the favour of the rodent during winter and spring. Lastly, I address the scatter-hoarding behaviour of A. subspinosus and cache recovery ability of its assumed closest food competitor Rhabdomys pumilio. I found that cache size has a profound influence on pilferage rates of L. sessile seeds. Acomys subspinosus scatter-hoarded the majority of seeds singly in the field and R. pumilio had difficulties finding those single-cached seeds in dry substrate under controlled experimental conditions, serving as evidence that scatter-hoarding is an effective method of pilferage mitigation by A. subspinosus during the dry summer months. Relatively little was known about this plant – disperser mutualism and how it functions before this thesis were conducted. I have provided insights into the influence of rodent disperser behaviour on seed morphology development, seed fate and seed persistence in the field and suspect that the plant –disperser relationship may have a larger influence on ecosystem dynamics than previously anticipated. Further research on this system is of importance, especially with today’s emerging environmental instability and human interference that threaten the robustness of highly interconnected ecosystems like the fynbos. / AFRIKAANSE OPSOMMING: Met die huidige omgewingskwessies, soos die klimaatsverandering, wat ekosisteme wêreldwyd affekteer, word die begrip van ekosisteemfunksionering toenemend belangrik. In hierdie studie ondersoek ek die dier – saadverspreidingsmutualisme tussen die endemiese verstrooiings-storing muis Acomys subspinosus en sy Proteaceae plant eweknie Leucadendron sessile in die biodiversiteit 'hotspot’ van die Kaapse Floristiese Ryk, Suid-Afrika. Die hoof doelwit van die tesis is om die saadseleksie en storingsgedrag van A. subspinosus te ondersoek. Eerstens het ek die saadseleksie strategie vir die verspreiding en begrawing deur A. subspinosus bepaal. Acomys subspinosus het direksionele druk uitgeoefen op L. sessile sade deur mediumgrootte sade met medium saadhuiddiktes te versprei en te begrawe. Klein sade was in situ geëet en groot sade was gelaat by afgesette plekke. Ek het die gevolgtrekking gemaak dat die L. sessile sade wat begrawe is ‘n kompeterende voordeel mag hê wanneer dit kom by die vestiging van saailinge in ‘n afgebrande omgewing, aangesien sade wat in die fynbos deur knaagdiere versprei word, soos L. sessile, baie groter is en dus meer gestoorde voedingstowwe en spoedige groeivermoëns het, as sade wat deur ander vektore versprei word. Tweedens het ek die knaagdier verspreidingsgedrag oor die tydperk van ‘n jaar gedokumenteer. Acomys subspinosus se verspreidingsgedrag het beduidend verander deur die verloop van die jaar, wat ek toegeskryf het aan die verandering in voedselbesbikbaarheid soos wat die jaar gevorder het. Acomys subspinosus het sade begrawe in die herfs na grootskaalse vrylating en val van die sade, maar het gestoorde sade begin terug kry en sade begin eet gedurende die winter en lente. Die knaagdier het na ‘n insekvretende dieët omgeskakel in die lente. Ek stel voor dat die A. Supspinosus – L. sessile verhouding nie die hele jaar mutualisties is nie, maar eerder antagonisties, in die knaagdier se guns, gedurende die winter en lente. In die laaste hoofstuk spreek ek die verstrooiings-storingsgedrag van A. subspinosus en storingsverkrygingvermoeë van sy naaste voedselmededinger en deponeringsdief Rhabdomys pumilio aan. Ek het gevind dat die storingsgrootte ‘n beduidende invloed het op die koers van diefstal van L. sessile sade. Acomys subspinosus het die meerderheid van die sade gestoor in die veld en R. pumilio het die enkel-gestoorde sade in droeë substraat onder gekontroleerde eksperimentele kondisies moeiliker gevind. Dit is ondersteunende bewyse dat verstrooings-storingsgedrag ‘n effektiewe metode is om diefstal te verminder in die droë somer in die fynbos. Relatief min was bekend oor hierdie dier – saad verspreidingsmutualisme en hoe dit funksioneer voordat die studie uitgevoer was. Ek het insig verskaf oor die invloed van knaagdier verspreidingsgedrag op saadmorfologie ontwikkeling, die lot van sade en die tydperk wat dit begrawe is in die veld. Ek vermoed dat die mutualisme ‘n hoeksteenproses is in die fynbos en die invloed daarvan op ekosisteemdinamieka mag dalk groter wees as wat voorheen verwag was. Verdere navorsing oor hierdie sisteem is belangrik, veral met vandag se opkomende omgewingsonstabiliteit en menslike inmenging wat die robuustheid van hoogs verbonde-netwerk ekosisteme soos die fynbos bedreig.

The present thesis aims to introduce process-based model for species range dynamics that can be fitted to abundance data. For this purpose, the well-studied Proteaceae species of the South African Cape Floristic Region (CFR) offer a great data set to fit process-based models. These species are subject to wildflower harvesting and environmental threats like habitat loss and climate change. The general introduction of this thesis presents shortly the available models for species distribution modelling. Subsequently, it presents the feasibility of process-based modelling. Finally, it introduces the study system as well as the objectives and layout. In Chapter 1, I present the process-based model for range dynamics and a statistical framework to fit it to abundance distribution data. The model has a spatially-explicit demographic submodel (describing dispersal, reproduction, mortality and local extinction) and an observation submodel (describing imperfect detection of individuals). The demographic submodel links species-specific habitat models describing the suitable habitat and process-based demographic models that consider local dynamics and anemochoric seed dispersal between populations. After testing the fitting framework with simulated data, I applied it to eight Proteaceae species with different demographic properties. Moreover, I assess the role of two other demographic mechanisms: positive (Allee effects) and negative density-dependence. Results indicate that Allee effects and overcompensatory local dynamics (including chaotic behaviour) seem to be important for several species. Most parameter estimates quantitatively agreed with independent data. Hence, the presented approach seemed to suit the demand of investigating non-equilibrium scenarios involving wildflower harvesting (Chapter 2) and environmental change (Chapter 3). The Chapter 2 addresses the impacts of wildflower harvesting. The chapter includes a sensitivity analysis over multiple spatial scales and demographic properties (dispersal ability, strength of Allee effects, maximum reproductive rate, adult mortality, local extinction probability and carrying capacity). Subsequently, harvesting effects are investigated on real case study species. Plant response to harvesting showed abrupt threshold behavior. Species with short-distance seed dispersal, strong Allee effects, low maximum reproductive rate, high mortality and high local extinction are most affected by harvesting. Larger spatial scales benefit species response, but the thresholds become sharper. The three case study species supported very low to moderate harvesting rates. Summarizing, demographic knowledge about the study system and careful identification of the spatial scale of interest should guide harvesting assessments and conservation of exploited species. The sensitivity analysis’ results can be used to qualitatively assess harvesting impacts for poorly studied species. I investigated in Chapter 3 the consequences of past habitat loss, future climate change and their interaction on plant response. I use the species-specific estimates of the best model describing local dynamics obtained in Chapter 1. Both habitat loss and climate change had strong negative impacts on species dynamics. Climate change affected mainly range size and range filling due to habitat reductions and shifts combined with low colonization. Habitat loss affected mostly local abundances. The scenario with both habitat loss and climate change was the worst for most species. However, this impact was better than expected by simple summing of separate effects of habitat loss and climate change. This is explained by shifting ranges to areas less affected by humans. Range size response was well predicted by the strength of environmental change, whereas range filling and local abundance responses were better explained by demographic properties. Hence, risk assessments under global change should consider demographic properties. Most surviving populations were restricted to refugia, serving as key conservation focus.The findings obtained for the study system as well as the advantages, limitations and potentials of the model presented here are further discussed in the General Discussion. In summary, the results indicate that 1) process-based demographic models for range dynamics can be fitted to data; 2) demographic processes improve species distribution models; 3) different species are subject to different processes and respond differently to environmental change and exploitation; 4) density regulation type and Allee effects should be considered when investigating range dynamics of species; 5) the consequences of wildflower harvesting, habitat loss and climate change could be disastrous for some species, but impacts vary depending on demographic properties; 6) wildflower harvesting impacts varies over spatial scale; 7) The effects of habitat loss and climate change are not always additive. / Das Ziel dieser Studie bestand daher darin, prozess-basierte Modelle zu entwickeln, die mit Daten zur Abundanz von Arten parametrisiert werden können. Die außergewöhnlich gut erforschten Proteaceen der südafrikanischen Kapregion (CFR), für die ein umfangreicher Datensatz zur Verfügung steht, stellen ein sehr geeignetes Untersuchungssystem zur Erstellung derartiger prozess-basierter Modelle dar. In Kapitel 1 beschreibe ich ein prozess-basiertes Modell für die Verbreitungsdynamik sowie die Methoden zur Parametrisierung des Modells mit Daten zu Abundanzverteilungen. Das Modell umfasst ein räumlich-explizites demographisches Modul und ein Beobachtungsmodul. Das demographische Modul verbindet artspezifische Habitatmodelle, die das geeignete Habitat beschreiben, und prozess-basierte demographische Modelle, die die lokale Dynamik und die Windausbreitung von Samen umfassen. Nach der Überprüfung der Parametrisierungs¬methoden mit simulierten Daten, wende ich die Modelle auf acht Proteaceenarten mit unterschiedlichen demographischen Eigenschaften an. Außerdem untersuche ich die Rolle von positiver (Allee-Effekte) und negativer Dichte-Abhängigkeit. Die Ergebnisse zeigen, dass Allee-Effekte und überkompensatorische Dynamik für viele Arten tatsächlich eine Rolle spielen. Der Großteil der geschätzten Parameter stimmt quantitativ mit unabhängigen Daten und beschreibt erfolgreich, wie die Abundanzverteilung aus der Bewegung und Interaktion der Individuen entsteht. Die vorgestellten Methoden scheinen daher zur Untersuchung von Ungleichgewichtsszenarien geeignet, die die Ernte von Infloreszenzen in Wildbeständen (Kapitel 2) und Umweltwandel (Kapitel 3) einschließen. In Kapitel 2 untersuche ich die Effekte der Ernte von Infloreszenzen in Wildbeständen. Das Kapitel beinhaltet eine Sensitivitätsanalyse über mehrere räumliche Skalen sowie demographische Eigenschaften. Darauf folgend wurden die Effekte der Ernte anhand von drei realen Arten untersucht. Die Reaktion der Pflanzen auf die Ernte zeigte ein Verhalten mit abrupten Schwellenwerten. Die durch die Ernte am stärksten gefährdeten Arten zeichneten sich durch kurze Samenausbreitungsdistanzen, starke Allee Effekte, geringe maximale Reproduktionsrate, hohe Mortalität und hohe lokale Aussterbewahrscheinlichkeit aus. Die Betrachtung größerer räumlicher Skalen wirkte sich trotz schärferer Grenzwerte positiv auf die Reaktion der Arten aus. Die drei untersuchten realen Arten konnten sehr geringe bis mittlere nachhaltige Ernteraten ertragen. Zusammenfassend lässt sich sagen, dass Kenntnisse über die Demographie des Untersuchungssystems und die umsichtige Identifizierung der zu betrachtenden räumlichen Skala zu einer besseren Einschätzung der Ernteintensität und der Naturschutzziele führen sollten. In Kapitel 3 wird die Reaktion der Arten auf vergangene Habitatverluste und zukünftigen Klimawandel sowie die Interaktion der beiden untersucht. Der Klimawandel wirkte sich dabei vornehmlich negativ auf die Größe des Verbreitungsgebiets und die Ausnutzung des potentiellen Habitats (‚Range Filling’) aus, wobei es zu einer Verschiebung des Habitats ohne erfolgreiche Kolonisierung kam. Der Habitatverlust reduzierte vor allem die lokalen Abundanzen. Die meisten Arten wurden vor allem durch das Szenario mit beiden Klimawandel und Habitatsverlust stark beeinträchtigt. Der negative Effekt war allerdings geringer als nach einer einfachen Aufsummierung der Einzeleffekte zu erwarten wäre. Dies erklärt sich aus einer Verschiebung des Verbreitungsgebiets der Arten in Regionen, in denen es in der Vergangenheit zu geringeren Habitatverlusten kam. Die Größe des Verbreitungsgebiets wurde am besten durch die Stärke des Umweltwandels vorhergesagt, wogegen das Range Filling und die lokalen Abundanzen hauptsächlich von den demographischen Eigenschaften abhingen. Aus diesen Ergebnissen lässt sich schließen, dass Abschätzungen des Aussterbensrisikos unter Umweltwandel demographische Eigenschaften einbeziehen sollten. Die meisten überlebenden Populationen waren auf Refugien reduziert, die im Fokus der Naturschutzmaßnahmen stehen sollten. Zusammenfassend zeigen die Ergebnisse, dass 1) prozess-basierte demographische Modelle für die Verbreitungsdynamik von Arten mit Daten parametrisierbar sind; 2) die Einbeziehung demographischer Prozesse die Modelle für die Verbreitung von Arten verbessert; 3) verschiedene Arten von unterschiedlichen Prozessen beeinflusst werden und unterschiedlich auf Umweltwandel und Beerntung reagieren; 4) Dichteregulierung und Allee-Effekte bei der Untersuchung der Verbreitungsdynamik von Arten berücksichtigt werden sollten; 5) die Ernte von Infloreszenzen in Wildbeständen, sowie Habitatverlust und Klimawandel für manche Arten katastrophale Folgen haben können, deren Effekte aber von den demographischen Eigenschaften abhängen; 6) der Einfluss der Beerntung in Abhängigkeit von der betrachteten räumlichen Skala variiert; 7) die Effekte von Habitatverlust und Klimawandel nicht additiv sind.

Flowers from members of the genus Leucadendron have colourful bracts and long vase life that make them highly desirable cut-flowers. Breeding programs based on interspecific hybridization would encounter difficulty if pre- or post-fertilization barriers exist in the distant crosses. Embryo rescue is one of the commonly used approaches to overcome post-fertilization barriers in wide hybridization. In this study, intersectional and intersubsectional hybridization of Leucadendron was attempted. Observation of pollen-pistil interactions revealed that post-zygotic rejection was the main reason for the incompatibility of the crosses, therefore embryo rescue was adopted and a protocol was developed to raise the hybrids. To better understand the genome structure in the genus, karyotypes of selected species were analyzed. Chromosome examination indicated that all (27) Leucadendron species examined were diploid and had a chromosome number of 2n = 26. The chromosomes were small in size and had predominantly median to submedian centromeres. The karyotypes of the species were rather symmetrical and seemed to be primitive according to Stebbins' karyotype classification. DNA based PCR-RFLP and RAMP markers were developed to identify Leucadendron hybrids at an early age. RAMP analysis showed more discrimination in identifying Leucadendron hybrids than did PCR-RFLP. The occurrence of PCR recombination also proved to be a troublesome issue when using the PCR-RFLP method, whereas the clarity of the interpretion of the RAMP method was not influenced by PCR recombination. Interspecific hybridization in a breeding program can provide valuable information on grouping of the species for systematic purposes. Regression analysis between cross success rate and cpDNA character difference revealed that there was a highly significant correlation between them. Patterns of success for intersectional hybridizations in Leucadendron were generally consistent with current taxonomic hypotheses regarding the sectional division of the genus. Success was generally lower for intersectional crosses than for intrasectional crosses.

Thesis (MScAgric)--University of Stellenbosch, 2005. / ENGLISH ABSTRACT: Quality of certain Leucospermum and Leucadendron cultivars after approximately 21 days shipping has been reported to be substandard due to ‘drying out’ of leaves and, in the case of Leucadendrons, involucral leaves. The nature of the symptoms of this ‘drying out’ and the conditions under which they form, viz. long exposures to low temperatures, has led us to hypothesize that these are symptoms of chilling injury (CI). Chilling injury, as far as we are aware, has not been documented on Leucospermums or Leucadendrons. Typical CI symptomology is discussed and shown for Leucospermum ‘Gold Dust’, ‘High Gold’ and ‘Succession’ and for Leucadendron ‘Chameleon’, ‘Laurel Yellow’ and ‘Safari Sunset’. The nature of CI symptoms for Leucospermums and Leucadendrons was generally membranous breakdown that manifested in some cases as a ‘water soaked’ appearance which, at a more advanced stage, was generally visible as ‘dried out’ patches on the leaves. In the case of the Leucadendrons CI was also visible on the immature involucral leaves which are more sensitive to chilling conditions than mature leaves. Dark discoloration of especially immature involucral leaves is also a symptom of CI. As water uptake of shoots with chilling injury is hindered the styles of the Leucospermums wilt. As can be expected, the lower the temperature below the threshold temperature and the longer the exposure the more severe the symptoms. CI was recorded on cut flower shoots of Leucospermum ‘Gold Dust’, ‘High Gold’, ‘Rigoletto’, ‘Succession’ and ‘Vlam’ after 21 and 24 days storage at 1ºC. After 24 days storage the chilling injury was more severe than after 21 days storage in most cases. Each cultivar was pulsed with 5 ml per stem of a 2% (w/v) sugar solution of either lactulose, sucrose, glucose, fructose or mannose before storage. After storage, CI was recorded on day 0, 3, 7 and 10 of the vase phase. Of the cultivars tested ‘Vlam’ and especially ‘Rigoletto’ were more prone to chilling injury development. ‘High Gold’ and ‘Vlam’ shoots were pulsed with 0 (control), 1.5, 3 or 4% (w/v) solutions of either mannose or fructose. The best control of CI for both cultivars was achieved with 1.5% (w/v) solution. Lower concentrations of mannose and fructose were tested on ‘High Gold’ shoots, with a 1% (w/v) solution giving the best control for both. At high concentrations signs of toxicity became evident directly after pulsing. ‘High Gold’ shoots were pulsed with 1% (w/v) solutions of mannose and fructose and sugar analyses were performed on shoots at different stages of storage and after 10 days in the vase. A slight increase in mannose and fructose was detectable in the stems of the shoots directly after pulsing but not in the leaves or the inflorescences. This is due to the low concentrations being used. The levels of all the carbohydrates decreased during the 21 days storage and more so during the vase phase of the flowering shoots. The fact that such low concentrations were effective in controlling chilling injury suggests that the sugars may have an effect other than on the osmotic potential. Cut ‘flower’ shoots of Leucadendron ‘Chameleon’, ‘Laurel Yellow’ and ‘Safari Sunset’ were stored for 14, 21 and 28 days, at 1º, 3º and 5ºC and CI development recorded during the subsequent 10 day vase phase. ‘Laurel Yellow’ and ‘Safari Sunset’ showed signs of chilling injury on the leaves after 28 days storage at 3ºC or lower and ‘Safari Sunset’ stored for 21 days developed chilling injury during the vase phase. Immature involucral leaves were more sensitive to chilling injury than leaves. CI increased with longer exposure times and lower storage temperatures for all three cultivars evaluated. ‘Chameleon’ was the most chilling tolerant of the cultivars up to 21 days. At 5ºC chilling injury was low irrespective of cold storage duration but longer exposures to 1º and 3ºC resulted in increased chilling injury development during the vase phase. All three cultivars were pulsed with 5 ml per stem of a 1% (w/v) solution of lactulose, sucrose, glucose, fructose or mannose and stored for 14, 21 and 28 days at 1ºC. The sugars reduced chilling injury on the leaves for ‘Safari Sunset’ when stored for 28 days and, to a lesser extent, in ‘Chameleon’. The sugars failed to reduce chilling injury of the involucral leaves of ‘Chameleon’ and ‘Laurel Yellow’ whereas there was some control especially after 28 days for ‘Safari Sunset’. In some cases the sugar pulse exacerbated chilling injury. Chilling injury generally increased rapidly after storage during the first three days in the vase and then at a lower rate for the next seven days. Leucadendron ‘Chameleon’, ‘Laurel Yellow’ and ‘Safari Sunset’ ‘cut flower’ shoots were pulsed with a 1% (w/v) glucose solution. Expressed on a dry weight basis, an increase in glucose concentration was not detected. The reduction in chilling injury of leaves by a sugar pulse is speculated, as for the Leucospermums, to be as a result of their presence in the apoplast and not the symplast and that their presence there protects the membranes against chilling conditions in some way. / AFRIKAANSE OPSOMMING: Die kwaliteit van sekere Leucospermum en Leucadendron kultivars na ongeveer 21 dae verskeping is waargeneem as substandaard as gevolg van die uitdroog van blare en, in die geval van Leucadendrons, die ‘involucral’ blare. Die aard van die simptome van hierdie uitdroging en die toestande waaronder dit plaasvind nl. lang periodes van blootstelling aan lae temperature, het ons tot die hipotese gebring dat hierdie simptome van koueskade is. Sover as wat ons bewus is, is koueskade nog nie gedokumenteer op Leucospermums of Leucadendrons nie. Tipiese koueskade simptomologie word bespreek en gewys vir Leucospermum ‘Gold Dust’, ‘High Gold’ en ‘Succession’ en vir Leucadendron ‘Chameleon’, ‘Laurel Yellow’ en ‘Safari Sunset’. Die koueskade simptome vir Leucospermums en Leucadendrons was oor die algemeen membraan afbraak wat ‘n water deurdrenkte voorkoms tot gevolg gehad het wat in ‘n meer gevorderde stadium sigbaar was as uitgedroogde kolle op die blare. In die geval van Leucadendrons was koueskade ook sigbaar op die onvolwasse ‘involucral’ blare wat meer sensitief is vir koue toestande as volwasse blare. Donker verkleuring van veral onvolwasse ‘involucral’ blare is ook ‘n simptoom van koueskade. Aangesien wateropname van stele met koueskade verhinder word, verwelk die ‘styles’ van die Leucospermums. Soos verwag kan word hoe laer die temperature onder die drempel temperatuur en hoe langer die blootstelling, hoe meer ernstig die simptome. Koueskade is aangeteken op gesnyde blomstele van Leucospermum ‘Gold Dust’, ‘High Gold’, ‘Rigoletto’, ‘Succession’ en ‘Vlam’ na 21 en 24 dae opberging by 1°C. Na 24 dae opberging was die koueskade meer ernstig as na 21 dae opberging in meeste gevalle. Elke kultivar het 5ml per steel van ‘n 2% (g/v) suiker oplossing van laktolose, sucrose, glucose, fruktose of mannose voor opberging opgeneem. Na opberging is koueskade aangeteken op dag 0, 3, 7 en 10. Van die kultivars wat getoets is, was ‘Vlam’ en veral ‘Rigoletto’ meer geneig tot koueskade ontwikkeling. ‘High Gold’ en ‘Vlam’ stele is geplaas in oplossings van 0 (kontrole), 1.5, 3 of 4 % (g/v) oplossings van mannose of fruktose. Die beste beheer van koueskade vir beide kultivars is deur die 1.5 (g/v) oplossing behaal. Laer konsentrasies van mannose en fruktose is getoets op ‘High Gold’ stele met ‘n 1% (g/v) mannose oplossing wat die beste beheer gegee het. Met hoë konsentrasies het tekens van toksisiteit sigbaar geword direk na opneem van die oplossing. ‘High Gold’ stele is geplaas in 1% (g/v) oplossings van mannose of fruktose en suiker analises is uitgevoer op stele by verskillende stadiums van opberging en na 10 dae in die vaas. ‘n Effense toename in mannose en fruktose is waargeneem in die stele van die blomme direk na opname van die oplossing, maar nie in die blare of die blomme nie. Dit is as gevolg van die lae konsentrasies wat gebruik is. Die vlakke van al die koolhidrate het afgeneem gedurende die 21 dae opberging en nog meer so gedurende die vaas periode van die blommende stele. Die feit dat sulke lae konsentrasies effektief is in die beheer van koueskade dui daarop dat die suikers ‘n effek het anders as op die osmotiese potensiaal. Snyblomme van Leucadendron ‘Chameleon’, ‘Laurel Yellow’ en ‘Safari Sunset’ is opgeberg vir 14, 21 en 28 dae, by 1º, 3º en 5°C en koueskade ontwikkeling is aangeteken gedurende die opvolgende 10 dae vaas periode. ‘Laurel Yellow’ en ‘Safari Sunset’ het tekens gewys van koueskade op die blare na 28 dae opberging by 3°C of laer en ‘Safari Sunset’ opgeberg vir 21 dae het koueskade ontwikkel gedurende die vaas periode. Onvolwasse ‘involucral’ blare was meer sensitief vir koueskade as die blare. Koueskade het toegeneem met langer blootstellingstye en laer opbergins temperature vir al drie kultivars geëvalueer. ‘Chameleon’ was die mees koueverdraagsaam van die drie kultivars tot op 21 dae. By 5°C was laag ongeag van die koue opberging tydperk, maar langer blootstellings aan 1º en 3°C het gelei tot toename in koueskade ontwikkeling gedurende die vaas periode. Al drie kultivars is voorsien met 5ml per steel van ‘n 1% (g/v) oplossing van lactulose, sucrose, glucose, fruktose of mannose en opgeberg vir 14, 21 en 28 dae by 1°C. Die suikers het koueskade verminder op die blare van ‘Safari Sunset’ wanneer opgeberg vir 28 dae en, tot ‘n mindere mate, in ‘Chameleon’. Die suikers het egter nie koueskade verminder van die ‘involucral’ blare van ‘Chameleon’ en ‘Laurel Yellow’ nie, waar daar egter wel in ‘n mate beheer was veral na 28 dae vir ‘Safari Sunset’. In sommige gevalle het die voorsiening van suiker die koueskade vererger. Koueskade het oor die algemeen vinnig toegeneem na opberging gedurende die eerste drie dae in die vaas en dan teen ‘n laer tempo vir die volgende sewe dae. Leucadendron ‘Chameleon’, ‘Laurel Yellow’ en ‘Safari Sunset’ snyblom stele is voorsien van ‘n 1% (g/v) glukose oplossing. Uitgedruk op ‘n droëmassa basis is ‘n toename in glukose konsentrasie nie waargeneem nie. Die afname in koueskade van blare deur die voorsiening van ‘n suiker oplossing is gespekuleer vir die Leucospermums, om ‘n resultaat te wees van hulle teenwoordigheid in die apoplas en nie die simplas nie, en dat die teenwoordigheid daar die membrane op ‘n manier beskerm teen koue toestande.