Made available in DSpace on 2014-06-11T19:33:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-12-12Bitstream added on 2014-06-13T19:04:54Z : No. of bitstreams: 1 almeida_ej_dr_jabo.pdf: 2925633 bytes, checksum: 1794e969fffc6c2ce4cdd74354b559fc (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Este trabalho teve os objetivos de estudar comparativamente os caracteres morfológicos e morfométricos úteis para distinção entre Meloidogyne mayaguensis e M. incognita; quantificar os danos causados à goiabeira ‘Paluma’, em formação, por M. mayaguensis em níveis crescentes de inóculo; caracterizar o desbalanço nutricional e a fenologia da goiabeira ‘Paluma’ face à infecção pelo nematóide; avaliar a hospedabilidade de outras frutíferas e plantas invasoras a esse patógeno; estudar a flutuação populacional do nematóide em pomar de goiabeira ‘Paluma’; prospectar materiais resistentes a M. mayaguensis dentre diversos acessos de Psidium guajava e Psidium spp. e avaliar a compatibilidade desses materiais como porta-enxerto para goiabeira ‘Paluma’. Os dados indicam que a morfologia do padrão perineal e da região labial dos machos são suficientes para a distinção segura entre M. mayaguensis e M. incognita. As medidas do diâmetro do tronco, comprimento das três pernadas, massa fresca e seca da parte aérea, retirada nas podas, área de folhas individuais, comprimento, largura, número por planta e massa dos frutos foram inversamente proporcionais aos níveis de inóculo de 0, 10, 100, 1.000 e 10.000 ovos e juvenis/planta. Os teores de N, P e K decresceram de folhas sem sintomas para folhas com o sintoma leve e severo, nas análises realizadas em fevereiro e agosto. / This work had the objectives of to study the morphologic characters and useful morfometric comparatively for distinction between Meloidogyne mayaguensis and M. incognita; to quantify the damages caused to the guava 'Paluma', in growing, for M. mayaguensis in growing levels of inocula; to characterize the nutritional imbalance and the fenology of the guava 'Paluma' face to the infection for the nematode; to evaluate the host suitability of other fruitful ones and its weeds to that pathogen; to study the population fluctuation of the plant parasitic nematode in guava orchard 'Pedro Sato'; resistant material prospect to M. mayaguensis between several accesses of Psidium guajava and Psidium spp. and to evaluate the compatibility of those materials as rootstock for guava 'Paluma'. The data indicate that the morphology of the perineal pattern and of the labial area of the males they are enough for the distinction it holds between M. mayaguensis and M. incognita. The measures of the diameter of the log, length of the three blanches, fresh and dry mass of the blanches, retreat in the pruning, area of individual leaves, length, width, number for plant and mass of the fruits was inversely proportional at the levels of inocula of 0; 10; 100; 1,000 and 10,000 eggs and juveniles/plant. The tenors of N, P and K decreased of leaves without symptoms for leaves with the light and severe symptom, in the analyses accomplished in February and August. The Ca had your concentration increased with the worsening of the symptoms, in February and August and Mg only in August. Sulfur had a larger concentration in leaves without symptoms, in February. In August, the leaves with severe symptoms presented larger concentrations of S.

Resumo: Este trabalho teve os objetivos de estudar comparativamente os caracteres morfológicos e morfométricos úteis para distinção entre Meloidogyne mayaguensis e M. incognita; quantificar os danos causados à goiabeira 'Paluma', em formação, por M. mayaguensis em níveis crescentes de inóculo; caracterizar o desbalanço nutricional e a fenologia da goiabeira 'Paluma' face à infecção pelo nematóide; avaliar a hospedabilidade de outras frutíferas e plantas invasoras a esse patógeno; estudar a flutuação populacional do nematóide em pomar de goiabeira 'Paluma'; prospectar materiais resistentes a M. mayaguensis dentre diversos acessos de Psidium guajava e Psidium spp. e avaliar a compatibilidade desses materiais como porta-enxerto para goiabeira 'Paluma'. Os dados indicam que a morfologia do padrão perineal e da região labial dos machos são suficientes para a distinção segura entre M. mayaguensis e M. incognita. As medidas do diâmetro do tronco, comprimento das três pernadas, massa fresca e seca da parte aérea, retirada nas podas, área de folhas individuais, comprimento, largura, número por planta e massa dos frutos foram inversamente proporcionais aos níveis de inóculo de 0, 10, 100, 1.000 e 10.000 ovos e juvenis/planta. Os teores de N, P e K decresceram de folhas sem sintomas para folhas com o sintoma leve e severo, nas análises realizadas em fevereiro e agosto. / Abstract: This work had the objectives of to study the morphologic characters and useful morfometric comparatively for distinction between Meloidogyne mayaguensis and M. incognita; to quantify the damages caused to the guava 'Paluma', in growing, for M. mayaguensis in growing levels of inocula; to characterize the nutritional imbalance and the fenology of the guava 'Paluma' face to the infection for the nematode; to evaluate the host suitability of other fruitful ones and its weeds to that pathogen; to study the population fluctuation of the plant parasitic nematode in guava orchard 'Pedro Sato'; resistant material prospect to M. mayaguensis between several accesses of Psidium guajava and Psidium spp. and to evaluate the compatibility of those materials as rootstock for guava 'Paluma'. The data indicate that the morphology of the perineal pattern and of the labial area of the males they are enough for the distinction it holds between M. mayaguensis and M. incognita. The measures of the diameter of the log, length of the three blanches, fresh and dry mass of the blanches, retreat in the pruning, area of individual leaves, length, width, number for plant and mass of the fruits was inversely proportional at the levels of inocula of 0; 10; 100; 1,000 and 10,000 eggs and juveniles/plant. The tenors of N, P and K decreased of leaves without symptoms for leaves with the light and severe symptom, in the analyses accomplished in February and August. The Ca had your concentration increased with the worsening of the symptoms, in February and August and Mg only in August. Sulfur had a larger concentration in leaves without symptoms, in February. In August, the leaves with severe symptoms presented larger concentrations of S. / Orientador: Jaime Maia dos santos / Coorientador: Antonio Baldo Geraldo Martins / Banca: José Antonio Alberto da Silva / Banca: Antonio Lúcio Mello Martins / Banca: Luiz Carlos Camargo Barbosa Ferraz / Banca: João Carlos de Oliveira / Doutor

A região da península Antártica possui uma plataforma profunda com características similares ao mar profundo e é uma região muito afetada pela variabilidade climática. Abaixo dos 60 m de profundidade, a meiofauna da baía do Almirantado e do talude adjacente no estreito de Bransfield é desconhecida. Este trabalho visa caracterizar a comunidade da meiofauna, gerando um conhecimento de base sobre sua biodiversidade e estrutura, relacionando com as variáveis ambientais (tipo de sedimento e quantidade/qualidade do alimento). Foi avaliado como a composição e a densidade da meiofauna variaram com a profundidade (100 e 1100 m), verticalmente no sedimento (0-6 cm) e entre dois anos com características climáticas contrastantes (dez/2008 e dez/2009). Também foi analisada a biomassa e diversidade dos Nematoda em nível específico em dez/2008. A densidade da meiofauna e a biomassa de Nematoda foram altas em comparação com outros locais. Houve grande variação na densidade da meiofauna entre os anos. Em dez/2008, ano mais quente, com pouca influência da cobertura de gelo e maior produtividade primária, a densidade variou com a profundidade, sendo alta dentro da baía, com maior concentração de fitodetritos, e baixa no estreito de Bransfield, onde o sedimento foi mais grosso indicando maior hidrodinamismo. Já em dez/2009, ano muito frio, com cobertura de gelo extensa e prolongada, com baixa produtividade primária não houve variação batimétrica da densidade ou das variáveis ambientais. A densidade e diversidade foram mais altas no estrato superficial do sedimento. Foram identificados 83 gêneros de Nematoda, que foram distinguidos em 185 morfoespécies, sendo que a maioria é espécie nova. A diversidade de espécies de Nematoda foi alta, como tem sido reportado para outras regiões antárticas e de mar profundo. A biomassa total de Nematoda foi cinco vezes mais alta que a média mundial, e variou entre as profundidades da mesma forma que a densidade. A biomassa individual, no estrato 0-2 cm, foi baixa na baía e alta no estreito. A morfometria mostrou que os indivíduos são mais longos e finos nas maiores profundidades, tanto batimétrica quanto vertical. A distribuição da meiofauna e da biomassa de Nematoda na área pareceu ser mais afetada por processos físicos (correntes, ressuspensão, topografia íngreme) do que pela quantidade de alimento disponível. / The Antarctic Peninsula has a deep shelf featuring similar conditions to the deep sea and it is highly affected by climatic variations. At Admiralty Bay and the adjacent slope in the Bransfield Strait, the meiofauna below 60 m depth is unknown. The present study aims to characterize the meiofauna community, and so generate a database on biodiversity and structure through the linkage to environmental variables (sediment type and quantity/quality of food). Composition and density of meiofauna have been evaluated across depth gradient (100 and 1100 m), sediment column (0-6 cm) and two years of contrasting climatic conditions (dec/2008 and dec/2009). Nematode biomass and diversity in specific level have been analyzed exclusively in the dec/2008 samples. Meiofauna density and nematode biomass were higher than in other localities. There was great variation of meiofauna density between the years. In dec/2008, density varied along the depth gradient: higher at the Admiralty Bay and lower at the Bransfield Strait, where the sediment was coarser (indicating high hydrodynamics). This year was the warmest, with weak influence of ice cover, high primary productivity and high phytodetritus concentration. Distinctively, in dec/2009, no bathymetric variation in meiofauna density or environmental variables was observed. This year was the coldest, with an extensive and enduring ice cover and low primary productivity. Density and diversity were higher in the superficial layer of the sediment. In total 83 nematode genera were found, comprising 185 morphospecies, mostly of new species. Species diversity of nematodes was elevated as reported in other Antarctic regions and in the deep sea. Nematode total biomass was five times higher than the global mean values, showing variation along the depth gradient. Individual biomass of layer 0-2 cm was low at the bay and high at the strait. Individuals were longer and thinner in deeper depths and sediment layers. Meiofauna and nematode biomass distributions seem to be more influenced by physical processes (currents, sediment ressuspension, steep topography) than by food availability.

Thesis (M.Sc. Agriculture (Soil Science)) -- University of Limpopo, 2019 / Nutritional water productivity (NWP) is an assessment tool, which describes the amount of water that has been used to produce selected mineral malnutrition (MMN) elements and micronutrient malnutrition (MNMN) substances. Therefore, it links agricultural production to human nutrition. Deficiencies in MMN elements and/or MNMN substances in human nutrition referred to as malnutrition, had been linked with fatal diseases. Agricultural soils could be affected by soil-borne pathogens such as plant-parasitic nematodes, which could limit the availability of MMN elements and MNMN substances. In some communities, vegetable crops, including chilli are regarded as a major source of MMN elements and MNMN substances. Effects of root-knot (Meloidogyne species) nematodes on NWP of chilli (Capsicum annuum L.) have not been documented. The objective of the study was to determine the effects of increasing population densities of M. incognita race 2 and M. javanica on the NWP of hot chilli plants. A microplot trial was conducted at the Green Biotechnologies Research Centre of Excellence (GBRCE), University of Limpopo, South Africa. Pots were filled with 10-L steam-pasteurised (300oC) sandy clay loam soil sourced from GBRCE and Hygromix-T (Hygrotech, Pretoria North) growth medium in the ratio 3:1 (v/v). Thereafter, three-week-old hot chilli cv. 'Serrano' seedlings were transplanted into each pot, with inoculum prepared by extracting eggs and second-stage juveniles (J2) of M. incognita race 2 and M. javanica from roots of grown nematode susceptible tomato cv. 'Floradade' (Solanum lycopersicum L.) in a 1% NaOCl solution. Fourteen days after transplanting, treatments 0, 50, 125, 250, 625, 1250 and 2000 eggs and second-stage juveniles (J2) of M. incognita race 2 and M. javanica were separately inoculated using a 20 ml plastic syringe into 5-cm-deep holes in pots. At 56 days after the initiation of the treatments, Meloidogyne species xiv decreased soil pH and increased organic carbon, contributing 29 and 43% in total treatment variation (TTV) of the respective variables. Treatment effects caused the pH to decrease. NWP variables against increasing nematode numbers exhibited quadratic relations, with coefficients of determination ranging from 59 to 86% for M. incognita race 2 trial and 80 to 98% for M. javanica trial. Meloidogyne species population densities against plant variables did not show any significant relationship, except for root galling and chlorophyll content where treatments contributed 76, 98 and 47% TTV of the respective variables. Generally, root galling increased with increase in Meloidogyne species population densities, whereas chlorophyll content decreased with increasing inoculum levels. Nematode variables against their increasing population exhibited quadratic relationship with the model explained by 44 to 95% for M. incognita race 2 and 28 to 82%, association, respectively for M. javanica. In conclusion, Meloidogyne species interfered with NWP of mineral elements in chilli plant and therefore, nematode management practices should be done to reduce the nematode population densities that would confer quality to agricultural produce for human health benefits.

The nematode Auanema rhodensis has recently been established as a new model organism. A. rhodensis is characterized by the simultaneous existence of three phenotypical sexes (males, females and hermaphrodites; called trioecy), skewed sex ratios which do not follow Mendel's laws, and variant segregation patterns according to sex and type of gametogenesis. Recently, A. rhodensis has been used to study the possible mechanisms for sex determination in three-sexed species and the variability of basic processes during sexual reproduction including meiotic divisions. During male meiosis, a diploid primary spermatocyte undergoes two consecutive divisions to form four haploid spermatids. Surprisingly, male meiosis in A. rhodensis results in two functional and two nonfunctional spermatids depending on the presence of an X-chromatid. A. rhodensis males exhibit a set of paired autosomes and one single X-chromosome, like males of the wellestablished nematode model organism Caenorhabditis elegans. In contrast to C. elegans, however, the X-chromosome in A. rhodensis divides precociously into its sister chromatids during the first meiotic division followed by a lagging X-chromatid and its uneven distribution during meiosis II. Additionally, the second meiotic divi-sion within this species is characterized by an asymmetric organelle distribution and a spindle structure reminiscent of a monopolar spindle. In this study, serial section electron tomography was used to analyse the ultrastructure of the microtubule skeleton in spermatocytes of A. rhodensis. The analysis of spermiogenesis using electron tomography posed some key advantages compared to standard transmission electron microscopy. First, the microtubule (MT) network could be studied in detail including spindle formation, organization of spindle poles, rearrangement of MTs, and inter-action between MTs and chromosomes. Second, the number and shape of chromosomes could be visualized. And third, the morphology of organelles could be observed at high resolution, and different organelles as well as their distribution pattern could be distinguished and quantified. This study provides highresolution 3D information about male meiosis in A. rhodensis. The results of this thesis confirm the complexity of the male meiotic program and the promi-nent position of the X-chromatid in meiosis II in this organism. Like previous light microscopic studies, electron tomography supports the hypothesis of an X-chromatid-dependent distribution of cellular organelles such as fibrousbody membranousorganelles (FB-MOs) and mitochondria. Furthermore, the formation of an asymmetric spindle could be observed with progressing anaphase II and might be associated with the X-chromatid distribution. Additionally, the analysis of the number of chromosome-associating MTs and their association character gives new insights into possible chromosome segregation mechanisms. Finally, significant differences to the male meiotic program in C. elegans have been identified. For the first time, the MT network in A rhodensis spermatocytes of different division stages has been observed in detail, and several different analyses could be done, including an analysis of the length distribution of MTs in the spindles. Because this ultrastructural analysis is based on fixed samples, live-cell imaging should be performed in the future to gain further information on the chromosome dynamics in this species. / Der Fadenwurm Auanema rhodensis hat sich in den letzten Jahren mehr und mehr zu einem neuen Modellorganismus entwickelt. Typisch für A. rhodensis ist das zeitgleiche Vor-kommen dreier phänotypischer Geschlechter (Männchen, Weibchen und Hermaphroditen; die sogenannte Triözie) und deren zahlenmäßig ungleiches Verhältnis zueinander, welches sich nicht durch Mendelsche Regeln erklären lässt. Ebenfalls auffällig sind nach Geschlecht und Gametogenese (Oogenese oder Spermatogenese) abweichende chromosomale Segregationsmuster. Unlängst hat A. rhodensis zu Erkenntnissen über die Geschlechtsdeterminierung in dreigeschlechtlichen Arten und die Varianz grundlegender Prozesse in der Meiose beigetragen. Während der männlichen Meiose (Spermatogenese) teilt sich eine diploide primäre Vorläuferzelle (primäre Spermatozyte) in zwei aufeinanderfolgenden Teilungen in insgesamt vier haploide Spermatiden. Bei A. rhodensis führt die Spermatogenese ungewöhnlicher-weise zu zwei funktionalen und zwei nicht-funktionalen Spermatiden, wobei die Funktionsfähigkeit vom Auftreten eines X-Chromatids abhängt. Männchen von A. rhodensis besitzen, ähnlich wie im Modellorganismus Caenorhabditis elegans, eine Reihe gepaarter Autosomen sowie ein einzelnes X-Chromosom. Im Gegensatz zu C. elegans teilt sich das ungepaarte X-Chromosom in A. rhodensis vorzeitig schon während der ersten meiotischen Teilung in seine Chromatiden, wodurch es zu einer verzögerten und ungleichen Verteilung des X-Chromatids während der zweiten meiotischen Teilung kommt. Diese zweite meiotische Teilung bei A. rhodensis ist außerdem durch eine asymmetrische Verteilung der Organellen und Mikrotubuli gekennzeichnet, letztere ähneln einer monopolaren Spindel. In dieser Arbeit wurde die Methode der seriellen Elektronentomographie genutzt, um die Ultrastruktur der Mikrotubuli in meiotischen Spindeln in Spermatozyten von A. rhodensis zu untersuchen. Zum einen wurden mittels Elektronentomographie das Netzwerk der Mikrotubuli und die Spindelorganisation, die Struktur der Spindelpole sowie die Interaktion zwischen Mikrotubli und Chromosomen drei-dimensional (3D) analysiert. Zum anderen wurde die Form der Chromosomen und die Morphologie und Verteilung der verschiedenen Organellen quantitativ erfasst. Somit stellt diese Studie hochauflösende 3D-Information über den Ablauf der männlichen Meiose in A. rhodensis zur Verfügung und bestätigt damit die Komplexität der männlichen Meiose und die zentrale Rolle des X-Chromatids während der zweiten meiotischen Teilung in diesem Organismus. Basierend auf vorangegangenen lichtmikroskopischen Experimenten an fixierten Proben unterstützt die Elektronentomographie die Hypothese einer vom X-Chromatid abhängigen Verteilung zellulärer Organellen wie spermienspezifischer FB-MOs oder Mitochondrien während der zweiten meiotischen Teilung. Außerdem konnte die Ausbildung einer asymmetrischen Spindel beobachtet werden, welche ebenfalls mit der ungleichen Verteilung des X-Chromatids in Zusammenhang stehen könnte. Eine zusätzliche Analyse chromosomenassoziierter Mikrotubuli brachte erste Erkenntnisse über mögliche zugrundeliegende Mechanismen der Chromosomensegregation. Die Ergebnisse dieser Arbeit konnten mit ähnlichen Untersuchungen in C. elegans verglichen und Unterschiede herausgearbeitet werden. Zum ersten Mal wurden hier meiotische Spindeln unterschiedlicher Teilungsstadien in 3D untersucht und unterschiedliche quantitative Analysen zur Längenverteilung der Mikrotubuli durchgeführt. Da alle hier gewonnenen Ultrastrukturdaten auf fixierten Proben basieren, sollte eine Betrachtung einer transgenen Wurmlinie mit Fluoreszenzmarkern mittels live-cell imaging auf diese Ultrastrukturanalyse folgen.

Meloidogyne incognita (Kofoid and White) is a major pest of soybean in South Africa and due to its high level of pathogenicity to the crop it is quintessential that research in this regard should receive priority. Root-knot nematode control has in the past mostly included the use of nematicides, while crop rotation and inclusion of cultivars with genetic host plant resistance (henceforth referred to as resistance only) to these pests were also used. Since no synthetically-derived and/or biological agents are registered locally as nematicides on soybean, the use of resistant cultivars represents one of the most viable and environmentally-friendly strategies to protect local soybean crops against damage resulting from parasitism by M. incognita. Although numerous exotic soybean cultivars have been identified with resistance to M. incognita, only a few locally adapted ones have proved to exhibit resistance to the latter species. Moreover, at present Egret is the only cultivar still available for commercial use in South Africa. Little and fragmented information is, however, available on the use of plant enzymes, that are interrelated in biochemical pathways that are expressed in root-knot nematode resistant cultivars, for its use as an additional parameter to exploit such a trait. Therefore, the present study was undertaken to identify M. incognita resistance in selected, locally adapted soybean cultivars by quantifying and exploiting the latter trait by using enzyme activities as an additional parameter. In addition, resistance to M. incognita in selected resistant soybean cultivars was also verified by means of histopathological studies to identify cellular changes associated with the trait. In the first part of the present study, 31 locally adapted soybean cultivars of which 23 were commercially available in the 2012 growing season were evaluated for resistance to M. incognita. The latter was done by means of traditional screening protocols for which M. incognita-gall rating, egg and second-stage juvenile as well as the reproductive factor data per root system for each cultivar screened were recorded. Two greenhouse experiments were subsequently conducted concurrently, one of which the abovementioned nematode parameters were recorded 30 and the other 56 days after inoculation. Reproduction factor values were used as the main criterium to identify M. incognita resistance in local soybean cultivars since it is considered as a more reliable parameter for this specific type of evaluations. Reproduction factor values equal to and lower than one, indicating resistance to the M. incognita population used in this study, were recorded only for cultivar LS5995, as well as seven pre-released GCI cultivars. These eight cultivars also had very low egg, as well as egg and second-stage juvenile counts per root system, all of which differed significantly from the susceptible control, as well as a number of other cultivars. Root gall indices, on the other hand, did not show consistent results in terms of the identification of the host status of the 31 cultivar screened during this study. Using reproduction factor values, local farmers can thus be supplied with information on the resistance of commercially-available soybean cultivars. Eventually, such M. incognita-resistant cultivars can be used to reduce population levels of this nematode pest in fields of producers and also as valuable germplasm sources in breeding programs to introgress/stack this trait in newly-developed soybean cultivars. The second part of the study aimed to verify and exploit M. incognita-resistance in soybean either identified as resistant or susceptible during the screenings experiments, using enzymatic activity as biochemical markers. Cultivar LS5995 was included as the resistant and Dundee as the susceptible standard. The activity of three enzymes, namely guaiacol peroxidase, lipoxygenase and catalase were recorded at different time intervals in roots and leaf samples of the latter cultivars, of both nematode-inoculated and nematode-free plants of each cultivar. Significant (P ≤ 0.05) increases in guaiacol peroxidase activity in leaf and root samples of the M. incognita-resistant cultivars GCI7 and LS5995 (inoculated with J2) were recorded 24 hours (h) after onset of the experiment. Use of this enzyme thus emanated as a useful parameter to identify soybean cultivars that exhibit resistance against M. incognita, especially in leaves, which could substantially reduce the time needed to screen cultivars. In terms of lipoxygenase activity recorded, substantial variation existed between the cultivars tested. The M. incognita-susceptible cultivar Egret was the only cultivar for which a significant (P ≤ 0.05) increase in lipoxygenase activity in the roots was evident 24 h after inoculation. However, during the 48 h sampling time, significant (P ≤ 0.05) differences in lipoxygenase activity were also recorded for the two M. incognita resistant cultivars GCI7 and LS5995. Although the increase in lipoxygenase activity for the susceptible cultivar Egret was unexpected, it may indicate that some level of resistance is present in the latter cultivar, which has in previous studies been identified as resistant to M. incognita. Other factors such as a different M. incognita populations used and temperature differences in greenhouse conditions that applied in this study compared to that for an earlier study may, however, serve as explanations for the latter differences in host status identification of cultivar Egret. In terms of catalase activity recorded in leaf samples of the M. incognita-resistant cultivar LS5995, substantial reductions of as much as 35.6 % were recorded for J2-inoculated plants compared to those of the J2-free control plants. In leaf samples of the susceptible cultivars, Egret and Dundee, catalase was also reduced, but to a lesser extent and ranged from 6 to 26 %. Conversely, catalase activity in the leaves of J2-inoculated plants of the highly susceptible cultivar LS6248R was substantially increased by as much as 29.3 %. Enzyme data obtained as a result of the current study thus generally complemented those of traditional screening assays in which resistance in locally adapted cultivars were identified to a certain degree. It is, however, recommended that enzyme activity, to be used as bio-markers, still needs further refinement and more investigation to optimise their use in identification, verification and exploitation of M. incognita resistance in soybean cultivars. The third and final part of the study encompassed a comparison of cellular changes induced by M. incognita in resistant and susceptible soybean cultivars to verify the resistant reactions expressed in the enzyme data. According to light- and transmission electron microscope observations, distinct differences in the appearance and development of giant cells in roots of the M. incognita-resistant cultivars LS5995 and GCI7 existed when compared to those in roots of the susceptible cultivars Dundee and LS6248R. In the latter cultivars, giant cells that formed were characteristically large and contained a dense cytoplasm, with thick irregularly surfaced cell walls. Cell walls also displayed thick aggregations that appeared to be cell-wall ingrowths. These giant cells are optimal to facilitate M. incognita development and reproduction. In contrast, giant cells that were associated with the resistant cultivars LS5995 and GCI7 were small, irregularly shaped and contained increased amounts of deposited cell-wall material in the cytoplasm known as cell wall inclusions. Necrosis was also present in M. incognita-infected root cells of both cultivars. Such giant cells have been associated with retarded feeding, development and reproduction of the latter root-knot nematode species. However, it was evident that neither GCI7 nor LS5995 are immune to M. incognita since J2 survived and developed to third- and fourth and ultimately mature females that reproduced in their roots. Optimal giant cells that were formed in the roots of the M. incognitasusceptible cultivars Dundee and LS6248R thus supported the nutritional needs of the developing M. incognita individuals and led to significant increases in M. incognita populations 56 days after inoculation as was evident from the high reproduction factor values that were obtained for such cultivars during host status assessments that represented the first part of this study. The opposite was recorded the M. incognita-resistant cultivars LS5995 and GCI7 since sub-optimal giant cells in their roots could not sustain high offspring from such mature females. The presence of necrotic root tissue adjacent to giant cells, furthermore, indicated that hypersensitive reactions occurred in the latter resistant cultivars. Enzyme data obtained in the second part of this study supported the presence of hypersensitive reactions in root cells of the latter resistant cultivars. Guaiacol peroxidase and lipoxygenase inductions in particular in plant tissues have been reported to play integral roles in hypersensitive reactions that are exhibited by cultivars that are resistant to pests and diseases. Finally, results obtained from the different parts of this study complemented each other. It resulted in the resistance that was identified in the GCI7 pre-released cultivar being verified and exploited against that of the resistant standard LS5995. Research that was done during this study also represented the first investigations into the use of enzymes as biochemical markers of resistance against M. incognita in soybean in South Africa. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Meloidogyne incognita (Kofoid and White) is a major pest of soybean in South Africa and due to its high level of pathogenicity to the crop it is quintessential that research in this regard should receive priority. Root-knot nematode control has in the past mostly included the use of nematicides, while crop rotation and inclusion of cultivars with genetic host plant resistance (henceforth referred to as resistance only) to these pests were also used. Since no synthetically-derived and/or biological agents are registered locally as nematicides on soybean, the use of resistant cultivars represents one of the most viable and environmentally-friendly strategies to protect local soybean crops against damage resulting from parasitism by M. incognita. Although numerous exotic soybean cultivars have been identified with resistance to M. incognita, only a few locally adapted ones have proved to exhibit resistance to the latter species. Moreover, at present Egret is the only cultivar still available for commercial use in South Africa. Little and fragmented information is, however, available on the use of plant enzymes, that are interrelated in biochemical pathways that are expressed in root-knot nematode resistant cultivars, for its use as an additional parameter to exploit such a trait. Therefore, the present study was undertaken to identify M. incognita resistance in selected, locally adapted soybean cultivars by quantifying and exploiting the latter trait by using enzyme activities as an additional parameter. In addition, resistance to M. incognita in selected resistant soybean cultivars was also verified by means of histopathological studies to identify cellular changes associated with the trait. In the first part of the present study, 31 locally adapted soybean cultivars of which 23 were commercially available in the 2012 growing season were evaluated for resistance to M. incognita. The latter was done by means of traditional screening protocols for which M. incognita-gall rating, egg and second-stage juvenile as well as the reproductive factor data per root system for each cultivar screened were recorded. Two greenhouse experiments were subsequently conducted concurrently, one of which the abovementioned nematode parameters were recorded 30 and the other 56 days after inoculation. Reproduction factor values were used as the main criterium to identify M. incognita resistance in local soybean cultivars since it is considered as a more reliable parameter for this specific type of evaluations. Reproduction factor values equal to and lower than one, indicating resistance to the M. incognita population used in this study, were recorded only for cultivar LS5995, as well as seven pre-released GCI cultivars. These eight cultivars also had very low egg, as well as egg and second-stage juvenile counts per root system, all of which differed significantly from the susceptible control, as well as a number of other cultivars. Root gall indices, on the other hand, did not show consistent results in terms of the identification of the host status of the 31 cultivar screened during this study. Using reproduction factor values, local farmers can thus be supplied with information on the resistance of commercially-available soybean cultivars. Eventually, such M. incognita-resistant cultivars can be used to reduce population levels of this nematode pest in fields of producers and also as valuable germplasm sources in breeding programs to introgress/stack this trait in newly-developed soybean cultivars. The second part of the study aimed to verify and exploit M. incognita-resistance in soybean either identified as resistant or susceptible during the screenings experiments, using enzymatic activity as biochemical markers. Cultivar LS5995 was included as the resistant and Dundee as the susceptible standard. The activity of three enzymes, namely guaiacol peroxidase, lipoxygenase and catalase were recorded at different time intervals in roots and leaf samples of the latter cultivars, of both nematode-inoculated and nematode-free plants of each cultivar. Significant (P ≤ 0.05) increases in guaiacol peroxidase activity in leaf and root samples of the M. incognita-resistant cultivars GCI7 and LS5995 (inoculated with J2) were recorded 24 hours (h) after onset of the experiment. Use of this enzyme thus emanated as a useful parameter to identify soybean cultivars that exhibit resistance against M. incognita, especially in leaves, which could substantially reduce the time needed to screen cultivars. In terms of lipoxygenase activity recorded, substantial variation existed between the cultivars tested. The M. incognita-susceptible cultivar Egret was the only cultivar for which a significant (P ≤ 0.05) increase in lipoxygenase activity in the roots was evident 24 h after inoculation. However, during the 48 h sampling time, significant (P ≤ 0.05) differences in lipoxygenase activity were also recorded for the two M. incognita resistant cultivars GCI7 and LS5995. Although the increase in lipoxygenase activity for the susceptible cultivar Egret was unexpected, it may indicate that some level of resistance is present in the latter cultivar, which has in previous studies been identified as resistant to M. incognita. Other factors such as a different M. incognita populations used and temperature differences in greenhouse conditions that applied in this study compared to that for an earlier study may, however, serve as explanations for the latter differences in host status identification of cultivar Egret. In terms of catalase activity recorded in leaf samples of the M. incognita-resistant cultivar LS5995, substantial reductions of as much as 35.6 % were recorded for J2-inoculated plants compared to those of the J2-free control plants. In leaf samples of the susceptible cultivars, Egret and Dundee, catalase was also reduced, but to a lesser extent and ranged from 6 to 26 %. Conversely, catalase activity in the leaves of J2-inoculated plants of the highly susceptible cultivar LS6248R was substantially increased by as much as 29.3 %. Enzyme data obtained as a result of the current study thus generally complemented those of traditional screening assays in which resistance in locally adapted cultivars were identified to a certain degree. It is, however, recommended that enzyme activity, to be used as bio-markers, still needs further refinement and more investigation to optimise their use in identification, verification and exploitation of M. incognita resistance in soybean cultivars. The third and final part of the study encompassed a comparison of cellular changes induced by M. incognita in resistant and susceptible soybean cultivars to verify the resistant reactions expressed in the enzyme data. According to light- and transmission electron microscope observations, distinct differences in the appearance and development of giant cells in roots of the M. incognita-resistant cultivars LS5995 and GCI7 existed when compared to those in roots of the susceptible cultivars Dundee and LS6248R. In the latter cultivars, giant cells that formed were characteristically large and contained a dense cytoplasm, with thick irregularly surfaced cell walls. Cell walls also displayed thick aggregations that appeared to be cell-wall ingrowths. These giant cells are optimal to facilitate M. incognita development and reproduction. In contrast, giant cells that were associated with the resistant cultivars LS5995 and GCI7 were small, irregularly shaped and contained increased amounts of deposited cell-wall material in the cytoplasm known as cell wall inclusions. Necrosis was also present in M. incognita-infected root cells of both cultivars. Such giant cells have been associated with retarded feeding, development and reproduction of the latter root-knot nematode species. However, it was evident that neither GCI7 nor LS5995 are immune to M. incognita since J2 survived and developed to third- and fourth and ultimately mature females that reproduced in their roots. Optimal giant cells that were formed in the roots of the M. incognitasusceptible cultivars Dundee and LS6248R thus supported the nutritional needs of the developing M. incognita individuals and led to significant increases in M. incognita populations 56 days after inoculation as was evident from the high reproduction factor values that were obtained for such cultivars during host status assessments that represented the first part of this study. The opposite was recorded the M. incognita-resistant cultivars LS5995 and GCI7 since sub-optimal giant cells in their roots could not sustain high offspring from such mature females. The presence of necrotic root tissue adjacent to giant cells, furthermore, indicated that hypersensitive reactions occurred in the latter resistant cultivars. Enzyme data obtained in the second part of this study supported the presence of hypersensitive reactions in root cells of the latter resistant cultivars. Guaiacol peroxidase and lipoxygenase inductions in particular in plant tissues have been reported to play integral roles in hypersensitive reactions that are exhibited by cultivars that are resistant to pests and diseases. Finally, results obtained from the different parts of this study complemented each other. It resulted in the resistance that was identified in the GCI7 pre-released cultivar being verified and exploited against that of the resistant standard LS5995. Research that was done during this study also represented the first investigations into the use of enzymes as biochemical markers of resistance against M. incognita in soybean in South Africa. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Master of Science / Department of Agronomy / William T. Schapaugh Jr / Fusarium virguliforme, the soilborne fungus which causes sudden death syndrome (SDS) of soybean, and Heterodera glycines Ichinohe, soybean cyst nematode (SCN), are two economically important pathogens in the Midwest. The pathogens are often found together in soybean (Glycine max (L.) Merr.) fields. This study was conducted to determine the effect of soybean genotype, F. virguliforme populations, and H. glycines populations have upon yield and to examine the interaction between the two pathogens. In 2008 and 2009, four genotypes with different levels of resistance to SDS and H. glycines were planted at seven environments. F. virguliforme and H. glycines soil populations were quantified at planting, midseason, and harvest. At the end of the growing season, area under the disease progress curves of SDS, F. virguliforme root populations, and H. glycines reproductive indices were determined and plots harvested for seed yield. Soil populations of F. virguliforme and H. glycines at planting, midseason, and harvest varied across environments. Within environments, generally, they were not significantly different. Seed yield varied within and across environments. As disease pressure increased, the performance of resistant genotypes increased compared to susceptible genotypes. Genotypes resistant to SDS yielded higher than susceptible genotypes. There were negative correlations between yield and disease rating and F. virguliforme root populations. F. virguliforme soil populations and H. glycines populations at planting were positively correlated. It is important to manage both SDS and H. glycines in fields with a history of the two diseases. This can be achieved through genetic resistance. Information in this study will improve decisions regarding genotype selection to minimize losses to SDS and H. glycines.

Master of Science / Department of Statistics / Leigh W. Murray / Traditionally the Poisson process is used to model count response variables. However, a problem arises when the particular response variable contains an inordinate number of both zeros and large observations, relative to the mean, for a typical Poisson process. In cases such as these, the variance of the data is greater than the mean and as such the data are over-dispersed with respect to the Poisson distribution due to the fact that the mean equals the variance for the Poisson distribution. This case study looks at several common and uncommon ways to attempt to properly account for this over-dispersion in a specific set of nematode count data using various procedures in SAS 9.2. These methods include but are not limited to a basic linear regression model, a generalized linear (log-linear) model, a zero-inflated Poisson model, a generalized Poisson model, and a Poisson hurdle model. Based on the AIC statistics the generalized log-linear models with the Pearson-scale and deviance-scale corrections perform the best. However, based on residual plots, none of the models appear to fit the data adequately. Further work with non-parametric methods or the negative binomial distribution may yield more ideal results.

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