Orientador: Gisela Ferreira / Resumo: Para que as plantas conseguissem conquistar o ambiente terrestre, foi necessário desenvolver habilidades que as tornassem capazes de sobreviver aos desafios de seu habitat, dentre as quais estão o desenvolvimento de mecanismos de tolerância à dessecação das sementes. Espécies que habitam locais com grande disponibilidade de água tendem a produzir sementes recalcitrantes e por outro lado, as espécies que habitam locais secos, como os Cerrados, tendem a produzir sementes ortodoxas, conseguindo tolerar períodos de seca sem acumular danos que prejudiquem a germinação. Dentre os sistemas que atuam no mecanismo de tolerância à dessecação estão a atividade de enzimas antioxidantes, o acúmulo de açúcares e a atividade das proteínas da embriogênese tardia (LEA). A família Annonaceae possui espécies inseridas em habitats contrastantes, como a Annona crassiflora, oriunda do Cerrado e a Annona glabra, oriunda de manguezais o que as torna interessantes para avaliar se suas sementes apresentam diferentes respostas frente à tolerância à dessecação. Para alcançar esse objetivo, sementes das duas espécies foram coletadas e avaliadas após submissão a diferentes níveis de secagem (teor inicial de água, 20%, 10% e 5%) e secagem seguida de reidratação. Foram realizados testes de germinabilidade, quantificação enzimática (superóxido dismutase (SOD) (EC 1.15.1.1), peroxidase (POD) (EC 1.11.1.7) e catalase (CAT) (EC 1.11.1.6)); de açúcares (frutose, galactose, glicose, manose, sacarose e trealose); ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In order for plants to conquer the terrestrial environment, it was necessary to develop skills that would enable them to survive the challenges of their habitat, including the development of seed desiccation tolerance mechanisms. Species that inhabit places with high water availability tend to produce recalcitrant seeds and, on the other hand, species that inhabit dry places, such as Cerrados, tend to produce orthodox seeds, being able to tolerate periods of drought without accumulating germinating damage. Systems that act on the desiccation tolerance mechanism include antioxidant enzyme activity, sugar accumulation, and late embryogenesis (LEA) protein activity. The Annonaceae family has species in contrasting habitats, such as Annona crassiflora, from the Cerrado and Annona glabra, from mangroves, which makes them interesting to evaluate if their seeds have different responses to desiccation tolerance. To achieve this goal, seeds of both species were collected and evaluated after submission to different drying levels (initial water content, 20%, 10% and 5%) and drying followed by rehydration. Germinability, enzymatic quantification (superoxide dismutase (SOD) (EC 1.15.1.1), peroxidase (POD) (EC 1.11.1.7) and catalase (CAT) (EC 1.11.1.6) tests were performed; sugars (fructose, galactose, glucose, mannose, sucrose and trehalose); determination of LEA proteins and lipoperoxide content. The results showed that A. crassiflora seeds were able to tolerate drying up to 10% water co... (Complete abstract click electronic access below) / Mestre

Uppsatsen behandlar hur de globala frösystemen kan ses som en typ av kolonisering av fröer. Uppsatsen utgår från Sarah Radcliffes definition av kolonialismen och menar att man inte endast kan se kolonialismen som en enskild händelse i tiden, utan att det är en pågående händelse som fortfarande äger rum och påverkar den värld vi lever i. Uppsatsen undersöker främst hur koloniseringen pågår i kunskapsproduktionen. Den gör sig tydligt i de globala frösystemen som är starkt påverkade av diskursen om modernitet som definierar vetenskaplig kunskap som modern och därför önskvärd, medan traditionella kunskaper ses som bakåtsträvande och därmed icke önskvärda. Detta visar sig i de globala frösystemen där allt fler stater övergår till ett jordbruk som är exportinriktat och där ”moderna” jordbruksmetoder används, på bekostnad av traditionella jordbruksmetoder som är anpassade till det lokala klimatet. Det gör att bönder hamnar i en utsatt situation då de ”moderna” jordbruksmetoderna kräver dyra resurser för att upprätthållas, samt att miljön degraderas då metoderna som nämndes inte är anpassade till den lokala miljön. Uppsatsen använder sig av kontraktsteori för att belysa hur frösystemen innebär en maktassymetri genom att de kontrakt som skrivs inom de globala frösystemen endast skrivs av en minoritet som sedan påverkar majoriteten som måste följa kontraktet. Därför krävs det att kontrakten inom de globala frösystemen skrivs om med en större inkludering och därmed ge utrymme för andra typer av kunskaper än endast den vetenskapliga. / This essay will discuss how the global seed systems can be viewed as a form of colonization of seeds. It will be based on Sarah Radcliffe’s definition of colonialism and suggests that you cannot only view colonialism as a separate event in time, but an on-going process still taking place, affecting the world we live in. The essay will mainly examine how the colonization is at work in the knowledge production. This is evident in the global seed systems that are strongly affected by the discourse of modernity that defines scientific knowledge as modern and therefore desired, while traditional knowledge is viewed as backwards and therefore non-desirable. This is showcased in the global seed systems where an increasing number of states move from an agriculture that is export-oriented and where “modern” agricultural methods are used, at the expense of traditional agricultural methods that are adapted to the local climate. This exposes the farmers to a situation where the “modern” agricultural methods demand expensive resources to maintain, while the environment suffers since the methods mentioned are not adapted to the local climate. The essay will make use of contract theory toilluminate how the seed systems means a power asymmetry through the contracts written within the global seed systems that are only written by a minority, which then affects the majority that must follow the contract. Therefore it is necessary for the contracts within the global seed systems to be rewritten, with a larger inclusion, and hence grant space for other forms of knowledge than merely the scientific.

Thesis (Ph. D. (Plant Production)) -- University of Limpopo, 2015 / Wild watermelon (Cucumis africanus LF.) and wild cucumber (Cucumis myriocarpus Naude.) are known for their ethnomedicine, ethnopesticide, ethnonematicide and nutritional properties, along with nematode resistance. The two Cucumis species were successfully used as inter-generic seedling rootstocks for watermelon (Citrullus lanatus Thunb.) cultivars, where nematode-resistant genotypes are not available. Also, the two Cucumis species are hardy and resilient to inland South Africa conditions, where temperatures are predicted to increase by 6°C in the year 2030. Seeds in the Cucurbitaceae Family contain high concentration of cucurbitacins, which induce auto-allelopathy that inherently inhibits plant growth and germination. Poor germination and non-uniform stands as a result of seed dormancy are a major challenge in sexual propagation of wild Cucumis species for various potential industries. Generally, true-to-type, uniform and disease-free plants in plant production are asexually-generated through in vitro propagation techniques. This study was therefore, initiated to address seed dormancy and related challenges of sexual propagation in the two wild Cucumis species by determining whether: (1) seed dormancy in C. africanus and C. myiocarpus would be ameliorated to allow for in vitro sexual propagation to establish pathogen-free parent stock, (2) the testa in C. africanus and C. myiocarpus seeds would possess structures, which interfere with imbibition and movement of water to the endosperm, (3) all organs of C. africanus and C. myriocarpus would be suitable for in vitro propagation, (4) suitable potting medium for in vitro propagated plantlets of C. africanus and C. myriocarpus would be available for acclimatisation of plantlets and (5) in vitro-produced xxviii plantlets from nematode-resistant C. africanus and C. myriocarpus would retain their resistance to Meloidogyne incognita race 2 under greenhouse conditions. In vitro and ex vitro experiments were conducted to achieve the stated objectives, with treatments in the laboratory and the greenhouse being arranged in completely randomised and randomised complete block designs, respectively. Validity was primarily ensured through the use of factorial trials, while the reliability of data was ensured by using appropriate levels of statistical significance. Leaching alone in C. africanus improved germination, while in C. myriocarpus this treatment had no effect on germination. The optimum leaching time in leached-control seeds of C. africanus was achieved at 7.1 h, with a 25-day mean germination time (MGT) and 52% optimum germination percentage (GP). In the two Cucumis species, the combined effect of leaching seeds in running tapwater and physical scarification of seeds at the chalaza region escalated germination in both Cucumis species, suggesting that both chemical and physical seed dormancies were involved. In C. africanus, cucurbitacin B (C32H48O8) was deposited exogenously to the testa, whereas in C. myriocarpus cucurbitacin A [cucumin (C27H4009) and leptodermin (C27H3808)], was deposited endogenously to the testa. The optimum leaching time in leached-scarified (LS) seeds of C. africanus was achieved at 5.7 h, with at least 40-day MGT and 89% optimum GP. In contrast, in C. myriocarpus LS seeds had the optimum leaching time of 6.3 h, with at least 41 days MGT and 93% optimum GP. Field emission SEM confirmed that there were two “water-gaps”, one at the micropylar region (hilum end) and the other at chalaza region (abaxial end) of seeds in both Cucumis species. Five distinct testa layers in seeds of C. myriocarpus were observed, namely, (i) epidermis, (ii) hypodermis, (iii) sclerenchyma, (iv) aerenchyma xxix and (v) chlorenchyma. In contrast, C. africanus seeds did not have the hypodermis between the micropylar and chalaza regions, but was present around both regions, which may provide some explanation of sporadic germination in non-leached and non-scarified seeds in this Cucumis species. The most suitable plant propagules for in vitro mass propagation of the two Cucumis species were nodal and apical buds. The optimum PGRs for shoot regeneration using both propagules in C. africanus and C. myriocarpus were at 0.80 and 0.35 μM 6-benzyladeninepurine (BAP), respectively. In contrast, the largest number of roots was regenerated at 0.31 and 0.44 μM indole-3-butyric acid (IBA) for C. africanus and C. myriocarpus, respectively. In vitro-produced plantlets were successfully acclimatised to ex vitro conditions, with sand + compost potting medium being the most suitable growing medium for weaning both Cucumis species. The in vitro-produced plantlets retained their resistance to M. incognita race 2. In conclusion, seeds of C. africanus and C. myriocarpus are structurally and chemically different, with strong evidence of chemical and physical dormancies. Structurally, C. myriocarpus seeds consist of five layers, four lignified and one non-lignified, whereas those of C. africanus have four layers, three lignified and one non-lignified. Evidence suggested that in C. africanus seeds, allelochemicals were primarily deposited outside the testa, whereas in C. myriocarpus they were deposited within the testa. The identified seed dormancies could successfully be ameliorated through combining leaching and scarification in both Cucumis species. The developed in vitro propagation protocols accord the two Cucumis species the potential for use as future crops in the context of climate-smart agriculture and research. / Flemish Interuniversity Council (VLIR)

>Magister Scientiae - MSc / Nature through billions of years of trial and error has produced an immeasurable amount of natural systems like plants, birds and animals. The intelligence of nature is hidden in these natural systems and researchers are turning towards “Nature’s intelligence” to find inspiration and advance novelty in the development of nanomaterials. Gold nanoparticles (AuNPs) have unique optical, electronic and physicochemical features which has gained them popularity and widespread exploitation in various applications. The conventional methods used for AuNPs synthesis employs toxic chemicals which makes these NPs unsafe for biomedical applications. Hence, there is a search for new, ‘green’ and more cost effective methods for AuNPs synthesis. Plant extracts are regarded as a highly desirable system for nanoparticle synthesis due to their tremendous capability to produce a wide range of phytochemicals that can act as reducing agents. The main goal of this study was to synthesize AuNPs in a cost effective manner without the use of toxic chemicals in the synthesis process. Avocado seeds which are an agricultural waste by-product were used for the biosynthesis of AuNPs. The study reports on the synthesis optimization, characterization and activities of the biogenic AuNPs. The avocado seed extract mediated - AuNPs (AvoSE-AuNPs) were optimized by varying reaction parameters and characterized by UV-visible, Dynamic Light Scattering (DLS) and High Resolution Transmission Electron Microscopy (HRTEM), Zetasizer and Fourier Transform Infrared Spectroscopy (FTIR). The formation of AvoSE-AuNPs had an absorption maximum at 534 nm. HRTEM and DLS confirmed that the NPs were polydispersed and present in different shapes. The presence of phytochemical constituents on the AvoSE-AuNPs were confirmed by FTIR. Their potential antibacterial activity was tested on bacterial strains known to exhibit resistance to a number of current antibiotics. The catalytic activity of AvoSE-AuNPs was also assessed as a means to contribute to the development of new methods aimed at alleviating organic pollutants such as nitrophenols in the environment. The AvoSE-AuNPs demonstrated excellent catalytic activity in the reduction of 4-NP by NaBH4 as shown by the rapid decrease in the nitrophenolate absorption band at 400 nm and the appearance of new absorption band at 298 nm, revealing the formation of the 4-aminophenol. Furthermore, the rate constants calculated demonstrated that the reaction occurs faster in the presence AvoSEAuNPs. The AvoSE-AuNPs showed low significant cytotoxicity. Cell cycle analysis was conducted to further investigate the apparent exhibited toxicity of the AvoSE-AuNPs. The results showed that in both cell lines treated with AvoSE-AuNPs and AvoSE there was a ii | P a g e disruption in the regulation of cell cycle. Cell cycle analysis helped improve understanding of the low cytotoxicity observed by the MTT assay results. The results presented in this study clearly demonstrate the feasibility of using AvoSE for the synthesis of AuNPs. This study demonstrated that AvoSE mediated AuNPs synthesis is a greener alternative as it abides by the green chemistry principles. Furthermore, the study outcomes contributed to minimizing environmental pollution by finding use for agricultural waste and thus ultimately adding value to the field.

<div>Fresh produce has been the leading source of foodborne illness outbreaks in the US, surpassing typical pathogen carriers such as meat, dairy, and seafood. Among the fresh produce popular to the consumers, cantaloupe and sprouts are mostly susceptible to pathogen contaminations and outbreaks. However, it has been a challenge to address the key factor in the contamination - the biofilms formed by pathogens are highly resistant to conventional washing and cleaning procedures. For cantaloupe, the net-like and porous surface forms a barrier for washing. For sprouts, the fragile texture of seedlings prevents aggressive cleaning operation and biofilm removal.</div><div><br></div><div>In this study, innovative interventions were developed to improve microbial safety of fresh produce, using cantaloupe and alfalfa sprouts as models. For cantaloupe, abrasive brushing was designed to remove pathogen biofilm from cantaloupe. Our research found pathogens could form biofilm at cantaloupe rind surface as the residence time of pathogens increased. Biofilm formed on cantaloupe rind was imaged by cryo-scanning electron microscopy (cryo-SEM), and its resistance to sodium hypochlorite and lauroyl arginate ethyl (LAE) was confirmed. Furthermore, abrasive brushing with peroxyacetic acid (PAA) could effectively remove biofilm formed at cantaloupe rind. The efficacy of this novel cleaning technique was highly desirable, which could achieve 3 log reduction in pathogen population. Mechanism of abrasive brushing to remove biofilm at cantaloupe rind surface was also proposed. Conceivably, brushing with diatomaceous earth (DE) and PAA could be an innovative and cost-effective method to remove pathogen biofilm from cantaloupe rind.</div><div><br></div><div>For alfalfa sprouts, since most of the outbreaks are linked to the sprouting seeds, seed disinfection treatments are considered to be the most effective method to improve microbial safety of sprouts. In this study, a newly developed alginate-based, antimicrobial seed coating treatment was evaluated for its efficacy to reduce foodborne pathogens from alfalfa seeds and sprouts. The calcium alginate coating in the presence of 2.5% lactic acid (CA-LA coating) reduced foodborne pathogens inoculated on alfalfa seeds to an undetectable level on day 1 during 28 day-seed storage, while chlorine (20,000 ppm) or lactic acid (2.5%) treatment took longer time to reach the same level. With sprouts, CA-LA coating resulted in > 2.5 log reduction for pathogen cells. In contrast, log reduction was < 0.6 for either chlorine (20,000 ppm) or lactic acid (2.5%) treatment. In general, this study indicated the effect of calcium alginate coating on reducing bacterial load of alfalfa seeds and sprouts, however, the germination rate of treated seeds was compromised due to the addition of lactic acid in the seed coating. Further study is needed to select antimicrobial compounds with minimum impact on germination rate of seeds.</div><div><br></div>

In common bean (Phaseolus vulgaris), the main seed storage pests are the bruchid beetles. Damage done to the seed by the larvae has a large impact on seed quality and yield. Arcelin (ARC), phytohaemagglutinin (PHA), and α-amylase inhibitor (α-AI) are linked seed storage proteins that form the APA locus on chromosome Pv04 and are associated with resistance. A major breeding objective is to introduce bruchid resistance into common bean from a resistant tepary genotype, G40199, by introgressing the resistant APA locus into susceptible common bean backgrounds. Here we developed a molecular marker that tracks the introgression. A set of PCR primers to the α-amylase inhibitor locus amplified a DNA fragment that showed a 45 base pair insertion in the middle of a lectin Leg_b domain. This enhanced locus characterization and insertion/deletion marker may preclude the need for bruchid resistance screening early in the breeding. / United States. Agency for International Development / United States. Global Hunger and Food Security Initiative (Cooperative Agreement No. EDH-A-00-07-00005-00)

Includes bibliographies. / The unique vegetation in the lowlands of the fynbos biome is threatened by alien Acacia encroachment. The seed bank dynamics of the two most widespread invaders in the region, Acacia saligma and A. cyclops, was studied to elucidate those factors contributing most to their invasive success. This information was then used to assist in developing optimal control methods. On the basis of information available prior to this study, it was predicted that both species would have large, persistent seed banks in the so.il, and that seed bank processes would provide the - key to invasive success: namely, high seed longevity and heat-stimulated germination. Seed banks were monitored for several years following clearing of the parent stand, using either sites sampled in an earlier study, or sites providing chronosequences of clearing dates. Concurrently, a demographic study of the species' seed banks, including processes from seed rain through to seedling emergence and survival, was done in dense Acacia stands and in fynbos vegetation. Acacia saligma seed banks conformed to predictions, being large and persistent owing to high percentage viability and water-impermeable dormancy. Seed banks accumulate rapidly under dense stands and are "disturbance-coupled" as they have potentially high longevity unless stimulated to germinate by fire. Although A. cyclops seed banks also may be large and long-lived, they display variable percentage viability and dormancy, with the majority of a seed cohort surviving less than a year. Acacia cyclops seed banks do not respond to heat treatment and appear to be "disturbance-uncoupled".

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This dissertation explores the differential effects of fires and rodent communities on native and invasive desert plant communities. Chapter one examines the impacts of fire and repeat fires on fuel loads in two different Utah desert sites, one in the cool Great Basin Desert and one in the hyper-arid Mojave Desert, over the course of four years. We found that both desert sites responded with varying intensities to a single burn, but the effects of a reburn were not as pronounced. We also found that our Great Basin Desert site had a stronger response to fire than our Mojave Desert site, producing a higher plant fuel loads that could potentially exacerbate the changing fire regimes. These data can be used to help map the effects that climate change may have on fuel loads and the fire potential of these deserts. Chapter two uses a full factorial experimental design to compare the effects of rodent herbivory and fire on native versus invasive seedlings at our Mojave Desert field site. We found that rodent herbivory has a more negative impact on the survival of native seedlings than invasive seedlings. This could be because the invasives are grasses that tend to tolerate herbivory better than the native shrubs and forbs. Chapter three again uses a full factorial experimental design to assess the impacts of rodents and fire on the fate of native and invasive seeds at our field site in the Mojave Desert. We found that rodents removed seeds, and they did show preferences for some species of seed over others, but these preferences were not different between native and invasive seeds or seed mass. We propose that the preferences may be influenced by other seed traits such as water content, handling time or physical and chemical defenses. As with the seedlings in chapter two, fire did not have any impact on rodent seed preferences. These data highlight the importance of considering rodent effects on seeds used in restoration effects following wildfires.

WRINKLED1 (AtWRI1) is a key transcription factor in the regulation of plant oil synthesis in seed and non-seed tissues. The structural features of WRI1 important for its function are not well understood. Comparison of WRI1 orthologs across many diverse plant species revealed a conserved 9 bp exon encoding the amino acids “VYL”. Site-directed mutagenesis of amino acids within the ‘VYL’ exon of AtWRI1 failed to restore the full oil content of wri1-1 seeds, providing direct evidence for an essential role of this small exon in AtWRI1 function. Arabidopsis WRI1 is predicted to have three alternative splice forms. To understand expression of these splice forms we performed RNASeq of Arabidopsis developing seeds and queried other EST and RNASeq databases from several tissues and plant species. In all cases, only one splice form was detected and VYL was observed in transcripts of all WRI1 orthologs investigated. We also characterized a phylogenetically distant WRI1 ortholog (EgWRI1) as an example of a non-seed isoform that is highly expressed in the mesocarp tissue of oil palm. The C-terminal region of EgWRI1 is over 90 amino acids shorter than AtWRI1 and has surprisingly low sequence conservation. Nevertheless, the EgWRI1 protein can restore multiple phenotypes of the Arabidopsis wri1-1 loss-of-function mutant, including reduced seed oil, the “wrinkled” seed coat, reduced seed germination, and impaired seedling establishment. Taken together, this study provides an example of combining phylogenetic analysis with mutagenesis, deep-sequencing technology and computational analysis to examine key elements of the structure and function of the WRI1 plant transcription factor.