Seeds of a range of crop species were classified according to their ability to germinate after presowing soaking treatments of varying severity. The differences in soaking tolerance were then compared with changes in several metabolic parameters to determine the importance of metabolic adaptation in the avoidance of soaking injury in seeds. After seed soaking treatments those species most susceptible to injury exhibited the highest respiration rates under nitrogen. At the same time the species with the highest respiration rates under nitrogen showed more rapid utilization of their seed reserves. This was demonstrated as a positive correlation of respiration rate under nitrogen against the fall in sucrose content of seeds on soaking. A similar positive correlation of respiration under nitrogen with alcohol content and with fall in sucrose content on soaking seeds was found. However, when the malic acid content of, seeds was assayed the reverse trend was shown, where malic acid was negatively correlated both with respiration under nitrogen and with the fall in sucrose content on soaking seeds. Thus the changes in the alcohol and malic acid contents of seeds on soaking were negatively correlated with each other. Analysis of lactic acid content of soaked seeds was expressed as the percentage of the three anaerobic products measured (ie. alcohol plus malic acid plus lactic acid) and gave a significant negative correlation against respiration rate under nitrogen' on soaking seeds. Parallel studies of both alcohol and malic dehydrogenases were conducted after soaking seeds. The activity of alcohol dehydrogenase was highest in those species which were least tolerant of soaking injury, while the Michaelis constants for alcohol dehydrogenase with respect to acetaldehyde were low in species intolerant of soaking injury but higher in species tolerant of this treatment. Enhanced metabolic damage was observed as a result of both high (30°C) and of low temperature (0°C) soaking treatments, but only in species which were normally susceptible to soaking injury. In studies on a single species (Pisum sativum) metabolic injury followed a similar course to the above studies in high but not in low temperature soaking treatments. Studies on the enzyme invertase showed grossly enhanced activity with a rise in temperature. This increased activity was also related to germination counts.

Bibliography: leaves 137-156. / xiv, 166 leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--Dept. of Horticulture, Viticulture and Oenology, University of Adelaide, 1993

Bibliography: leaves 137-156.

Desiccation-sensitive seeds show differential viability characteristics during drying at different rates. A number of studies have demonstrated that rapid dehydration permits survival to lower water contents than does slower desiccation. The aim and objective of the present study was to test the hypothesis which states that rapid drying of desiccation-sensitive seeds removes water sufficiently fast to reduce the accumulation of metabolic damage. In addition, the hypothesis that wet storage subjects desiccation-sensitive seeds to mild, but increasingly severe, water stress causing oxidative damage if additional water is not supplied, was tested. In the present study, axes of germinating orthodox seeds of Pisum sativum and newlyshed recalcitrant counterparts of Quercus robur, Strychnos madagascariensis, Trichilia emetica, Trichilia dregeana and Avicennia marina were subjected to rapid or slow drying or wet storage. For those species where more than one harvest was investigated, differences were observed in water contents at shedding. For all the species studied, the dehydration rate could be described by an exponential and a modified inverse function for both desiccation regimes, and the water content remained constant with wet storage. The level of tetrazolium staining and germination percentage of axes decreased sharply drying and hydrated storage such that the marked decline took place at lower water contents upon rapid than slow dehydration. The conductivity of electrolyte leachate increased progressively during desiccation and moist storage of axes of all species investigated. Greater membrane leakage occurred upon slow, than rapid dehydration in axes of all species studied. Activities of respiratory enzymes which have a potentially regulatory role in glycolysis, phosphofructokinase (PFK), or the tricarboxylic acid cycle, malate dehydrogenase (MDH), and levels of the oxidized form of the coenzyme, nicotinamide adenine dinucleotide (NAD), of the enzymes of the electron transport chain, NADH dehydrogenases ofNADH-ubiquinone (coenzyme Q) reductase (complex I) and NADHcytochrome c reductase (complex IV), were monitored in the present investigation. v In addition, the role of free radical activity in the form of lipid peroxidation, which has been implicated in loss of viability in seeds, was examined by assaying the levels of hydroperoxides. The involvement of the free radical processing enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR), and the antioxidant, ascorbic acid (AsA), was also ascertained. The activity of PFK in axes of P. sativum remained constant during drying and wet storage. However, PFK activity increased as rapid dehydration and hydrated storage of Q. robur axes proceeded. In contrast, the activity of PFK in axes of Q. robur decreased during slow desiccation. Similarly, PFK activity was reduced upon drying, and moist storage, of T. dregeana axes such that higher activity of PFK was seen during rapid than slow dehydration. The activity ofPFK inA. marina axes also declined upon desiccation. The activity ofMDH in axes of P. sativum was also unchanged during drying and wet storage. However, an increase in MDH activity was recorded in Q. robur axes during dehydration and hydrated storage such that the activity of MDH was higher upon slow than rapid desiccation. In contrast, MDH activity in axes of T. dregeana decreased as drying proceeded. Similarly, the activity of J\.1DH declined during dehydration and moist storage of A. marina axes. An increase in the level of NAD occurred in axes of P. sativum during drying. In contrast, a decrease in NAD levels was seen upon dehydration and wet storage of Q. robur axes such that the level of NAD was higher upon rapid than slow desiccation. There was an enhancement of the level of NAD in axes of T. dregeana during hydrated storage. Conversely, NAD levels declined during drying ofA. marina axes. A decrease in the level of hydroperoxides in axes of P. sativum was seen as rapid drying proceeded. In contrast, hydroperoxide levels increased during wet storage of P. sativum axes. Similarly, the levels of hydroperoxides were enhanced upon dehydration and hydrated storage of Q. robur axes such that they were higher in axes during slow desiccation compared to those dried rapidly. Conversely, the hydroperoxide level in axes of T. dregeana was reduced upon rapid dehydration. In contrast, an elevation of the level of hydroperoxides was observed during moist storage. The levels of hydroperoxides remained constant as desiccation and wet storage ofA. marina axes proceeded. vi The activity of SOD in axes of P. sativum decreased during rapid drying. In contrast, SOD activity increased upon slow dehydration and wet storage ofP. sativum axes. There was a decline in the activity of SOD in Q. robur axes during slow desiccation. Similarly, SOD activity was diminished upon drying of axes of T. dregeana. The activity ofSOD in T. dregeana axes was enhanced during hydrated storage. An elevation in SOD activity also took place during rapid dehydration and moist storage of axes ofA. marina. The activity of CAT did not change during drying of axes of P. sativum. However, a decrease in CAT activity in Q. robur axes was seen upon slow dehydration and wet storage. Similarly, the activity of CAT declined as desiccation of axes of T. dregeana proceeded. In contrast, CAT activity inA. marina axes increased during slow drying. Whereas the activity of GR in axes of P. sativum increased during drying and wet storage, GR activity decreased in A. marina axes upon all treatments such that the activity ofGR was higher during rapid than slow dehydration. GR activity also declined upon slow desiccation and hydrated storage ofaxes of Q. robur. Similarly, the activity of GR in T. dregeana axes was reduced during moist storage. Finally, a decrease in the level of AsA in axes of P. sativum took place during drying. Nonetheless, dehydration and wet storage of Q. robur axes were associated with no siginificant change in AsA levels. There was also a decline in the level of AsA in axes of T. dregeana as rapid desiccation proceeded. Similarly, a reduction in AsA level occurred upon slow drying ofaxes ofA. marina. The results presented here are consistent with the observation that drying and wet storage adversely affected the respiratory enzymes, PFK, MDH and NADH dehydrogenase. It is suggested that the resultant metabolic imbalance led to more leakage of electrons from the mitochondrial electron transport chain than normal, and through lipid peroxidation increased levels of hydroperoxides. In addition, dehydration and hydrated storage may depress the activities of free radical processing enzymes, SOD, CAT and GR and levels of antioxidant, AsA. This phenomenon was less pronounced during rapid, in comparison to slow, desiccation and moist storage. However, it appears that the above biochemical events are overtaken by physical damage at higher water contents in the highly recalcitrant seeds. It was concluded that the differential effects of VII the drying rate and wet storage on responses of desiccation-sensitive seeds varies with tissue, harvest, species and the degree of desiccation sensitivity. / Thesis (Ph.D.)-University of KwaZulu-Natal, 2004.

Biochemical and ultrastructural changes associated with chilling injury (CI) in soybean seeds imbibed at 5°C and 25°C were investigated. Soybean seed germination appeared to be affected by chilling temperatures and initial seed moisture content. Seeds with higher moisture contents exhibited 85% germination, while low moisture content seeds had a 32% germination. Leakage rates were greater in chilled seeds, indicating that membrane integrity in the tissues was impaired at chilling. The low rates of potassium ion leakage between 6 and 24 hours of imbibition compared to the high peroxide levels observed during this period led to the suggestion that lipid peroxidation was a better marker of CI than leakage. Transient changes in lipid hydroperoxide levels were observed in chilled and non-chilled seeds and axes. However, in axes, the increase in lipid hydroperoxides after 12 hours of imbibition at chilling temperatures was associated with an 18% decline in linoleic acid levels of total lipid fraction. Similarly, a 10% decline was observed in the polar lipid fraction. These results suggest that the capacity of seeds to control lipid peroxidation may be an important component in CI and that a consequence of peroxidation is likely to be a loss of fatty acid unsaturation. Sugar levels were not affected by chilling and non-chilling temperatures and no relationship could be established with CI. Antioxidant defense enzymes (catalase and superoxide dismutase) were expressed at chilling and non-chilling temperatures and increases were observed after 24 hours of imbibition which showed an apparent correlation with increases to lipid hydroperoxide levels. Enzyme levels decreased after 48 hours of imbibition at a time which coincided with the decline observed in the peroxide levels. Overall, no marked differences were observed in chilled and non-chilled cells at the ultrastructural level, except that vacuolar reserve mobilization was markedly impeded. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1996.

Control of seed-associated micro-organisms is vital in reducing losses of plants of economic importance. Recalcitrant seeds being metabolically active and able to be stored only under conditions of high relative humidity makes it more difficult to control contaminants. Nevertheless, means need to be developed to eliminate, or at least curtail, seed-associated fungi and bacteria. The use of biological control is a highly recommended alternative to chemical control for reducing the risk of killing beneficial organisms, as well as in terms of health and environmental hazards. Furthermore, when working with seed-derived tissues, it is extremely important to optimise a method or methods to control contamination without compromising the viability or further development of the explants. The original aim of the present study was to determine whether the biocontrol agents, EcoT® and Eco77® (commercial products of the spores of Trichoderma harzianum) would effectively control/eliminate micro-organisms from the embryonic axes of Trichilia dregeana, while promoting growth under in vitro conditions. Other means were also tested for their efficacy in controlling contaminants; these were application of Benlate®, Nipastat® (a mixture of parabens), anodic water (the anodic fraction of an electrolysed dilute solution of calcium and magnesium chloride), sodium dichloro-isocyanurate (Medi-Chlor®[NaDCC]) and alginate gel encapsulation of the embryonic axes. Prior to the experiments, fungal contaminants from the embryonic axes were isolated on potato dextrose medium and identified using light microscopy. EcoT and Eco77 were initially individually tested by co-culture as conidial suspensions with the embryonic axes. A further approach used liquid culture (potato dextrose broth) as well as solid culture medium (based on sugarcane bagasse) in/on which the strains of Trichoderma harzianum had been grown. This was aimed at testing for the possible presence of compounds released by T. harzianum into the media, which might prove to be effective in curtailing/eliminating the axis-associated microflora. Among the different treatments tested, the best method was utilised to decontaminate the embryonic axes prior to minimal-growth storage (hydrated axes encapsulated in alginate gel ‘beads’). Penicillium spp. were predominant among the different fungi isolated, which included Fusarium spp., Rhizopus spp., Aspergillus niger and Aspergillus flavus. Co-culturing with T. harzianum for 24 h was successful in terms of the survival of the embryonic axes, although the roots produced were shorter than when axes were cultured alone, but had no effect in eliminating the contaminants. Longer periods of co-culturing with T. harzianum affected the germination of the embryonic axes of T. dregeana compared with axes germinated in the absence of the biocontrol agent (control). The culture filtrate negatively affected germination of the T. dregeana embryonic axes, although it was effective against the associated contaminants. Nipastat was effective in reducing the contamination, and, depending on the concentration, did not affect germination adversely. Medi-Chlor was highly effective in eliminating all the contaminants from axes in vitro. Both these treatments were therefore used to decontaminate axes before minimal-growth storage. All the NaDCC-treated, encapsulated axes examined after 14 d hydrated storage [in Magenta boxes] and after 14-42 d in polythene bags survived; however the axes stored in aluminium foil-lined bags and Eppendorf tubes soon lost viability. The recommendation is therefore made that the decontamination treatment based on use of NaDCC (or other preparations of sodium dichloro-isocyanurate) be tested on embryonic axes of a range of recalcitrant-seeded species, and, if successful, the procedure be introduced into cryopreservation protocols. The use of NaDCC has emerged as a promising method of eliminating contaminating microflora which otherwise compromise in vitro procedures, from seed-derived explants. Furthermore, containment of decontaminated encapsulated axes in sealed polythene bags offers an apparently ideal means of temporary storage and dissemination. The results should find considerable applicability when excised embryonic axes representing the germplasm of recalcitrant seeds, are cryoconserved. / Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2013.

Made available in DSpace on 2014-06-11T19:22:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-10-19Bitstream added on 2014-06-13T18:48:43Z : No. of bitstreams: 1 dourado_ws_me_botfca.pdf: 306085 bytes, checksum: 1b0f493f9f26322e6c5f3a7602ed38d5 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / A ervilha forrageira difere da ervilha comum quanto a cor do tegumento, ao elevado teor de aminoácidos e foi selecionada visando a alimentação animal. O objetivo do trabalho foi verificar a eficiência de testes de vigor, com ênfase no envelhecimento acelerado para a avaliação da qualidade de sementes de ervilha forrageira visando seleção de lotes para a emergência de plântulas em campo e longevidade no armazenamento. Foram utilizados 10 lotes de ervilha forrageira avaliados quanto ao teor de água, massa de mil sementes, teste da primeira contagem, germinação, condutividade elétrica, emergência de plântulas em campo e teste de envelhecimento acelerado com as seguintes variações de metodologia: temperaturas de 41 e 42 °C; tempos de exposição das sementes de 48 e 72 horas e umidade da atmosfera de envelhecimento, mediante a utilização de água destilada (100% UR), solução saturada de cloreto de potássio (87% UR) e solução saturada de cloreto de sódio (76% UR). Os resultados encontrados foram comparados à emergência de plântulas em campo e armazenamento pelos tempos de 90, 180, 270, 360, 450, e 540 dias. Os testes de envelhecimento acelerado com água à 41° C por 48 horas, cloreto de potássio a 41 °C por 72 horas e 42 °C por 48 horas: são promissores para avaliar o vigor de lotes de sementes de ervilha forrageira fornecendo informações semelhantes à emergência de plântulas em campo. Nenhum das variáveis do teste de envelhecimento avaliadas mostrou eficiência na seleção de lotes quanto ao desempenho no armazenamento / The pea pea differs common as the color of the coat, the high content of amino acids and was selected in order to feed. The aim of the study was to assess the efficiency of vigor tests, with emphasis on accelerated aging to assess the quality of pea seeds aimed selection of lots for field seedling emergence and longevity in storage. We used 10 batches of pea evaluated for water content, weight of thousand seeds, test the first count, germination, electric conductivity, seedling emergence in field and accelerated aging test with the following variations in methodology: temperatures of 41 and 42 °C; seeds exposure times of 48 and 72 hours and humidity aging, using distilled water (100% UR), saturated potassium chloride (87% UR) and saturated chloride sodium (76% UR). The results were compared to seedling emergence in field and storage for times of 90, 180, 270, 360, 450, and 540 days. The accelerated aging tests with water at 41 °C for 48 hours, potassium chloride at 41 °C for 72 hours and 42 °C for 48 hours: are promising to evaluate the effect of seed lots of pea similar information to seedling emergence in the field. None of the variables evaluated aging test showed efficiency in selecting lots for performance in storage

Orientador: Cibele Chalita Martins / Banca: Carla Gomes Machado / Banca: Sandra Aparecida de Souza / Resumo: A ervilha forrageira difere da ervilha comum quanto a cor do tegumento, ao elevado teor de aminoácidos e foi selecionada visando a alimentação animal. O objetivo do trabalho foi verificar a eficiência de testes de vigor, com ênfase no envelhecimento acelerado para a avaliação da qualidade de sementes de ervilha forrageira visando seleção de lotes para a emergência de plântulas em campo e longevidade no armazenamento. Foram utilizados 10 lotes de ervilha forrageira avaliados quanto ao teor de água, massa de mil sementes, teste da primeira contagem, germinação, condutividade elétrica, emergência de plântulas em campo e teste de envelhecimento acelerado com as seguintes variações de metodologia: temperaturas de 41 e 42 °C; tempos de exposição das sementes de 48 e 72 horas e umidade da atmosfera de envelhecimento, mediante a utilização de água destilada (100% UR), solução saturada de cloreto de potássio (87% UR) e solução saturada de cloreto de sódio (76% UR). Os resultados encontrados foram comparados à emergência de plântulas em campo e armazenamento pelos tempos de 90, 180, 270, 360, 450, e 540 dias. Os testes de envelhecimento acelerado com água à 41° C por 48 horas, cloreto de potássio a 41 °C por 72 horas e 42 °C por 48 horas: são promissores para avaliar o vigor de lotes de sementes de ervilha forrageira fornecendo informações semelhantes à emergência de plântulas em campo. Nenhum das variáveis do teste de envelhecimento avaliadas mostrou eficiência na seleção de lotes quanto ao desempenho no armazenamento / Abstract: The pea pea differs common as the color of the coat, the high content of amino acids and was selected in order to feed. The aim of the study was to assess the efficiency of vigor tests, with emphasis on accelerated aging to assess the quality of pea seeds aimed selection of lots for field seedling emergence and longevity in storage. We used 10 batches of pea evaluated for water content, weight of thousand seeds, test the first count, germination, electric conductivity, seedling emergence in field and accelerated aging test with the following variations in methodology: temperatures of 41 and 42 °C; seeds exposure times of 48 and 72 hours and humidity aging, using distilled water (100% UR), saturated potassium chloride (87% UR) and saturated chloride sodium (76% UR). The results were compared to seedling emergence in field and storage for times of 90, 180, 270, 360, 450, and 540 days. The accelerated aging tests with water at 41 °C for 48 hours, potassium chloride at 41 °C for 72 hours and 42 °C for 48 hours: are promising to evaluate the effect of seed lots of pea similar information to seedling emergence in the field. None of the variables evaluated aging test showed efficiency in selecting lots for performance in storage / Mestre