The causative agent of malignant tertian malaria, Plasmodium falciparum undergoes an arrested growth phenotype of its erythrocytic stage when under drug-stress. Recent artemisinin treatment failures seem to be indicative of such induction followed by recrudescence rather than actual therapeutic failure. Likewise, P. vivax hypnozoites are the prototypic dormants and the latent infections for which they are responsible prove most difficult to treat. Dihydroartemisinin, an artemisinin-derivative, can be used to exploit this mechanism by inducing a dormant state in ring-stage P. falciparum parasites and in turn, their recovery may be used as a screening period for compounds that inhibit or foster growth. Specifically, parasites stably transfected with luciferase were used to quantitatively observe growth (or lack thereof) response of parasites to the phytohormone gibberellic acid and the herbicide, fluridone. Using their behavior as comparative controls, the Medicines for Malaria Venture (MMV) Malaria Box was screened for similar activity. The most active compound, 1,2,3,4-tetrahydroacridin-9-ol a quinoline-derivate caused cells to wake even earlier than expected. Since quinine and other such drugs have historically been most effective in treating malaria, it seems appropriate that such a finding was made. Following this the MMV Box was screened again against uninduced 3D7 parasites to determine if any were capable of causing a dormant response under the hypothesis that such a reaction is a defensive adaptation of P. falciparum. Four compounds were found to be active of which two appear to be inducing dormancy in the second cycle rather than the first akin to DHA. These quiescent periods also appear to be shorter indicating that the latter is more efficient. It is possible that given the length of interaction with artemisinin, P. falciparum is more adept to respond to its derivatives likewise the mechanism of action may be different enough to change the nature of the response.

There is a discrepancy in the current literature concerning the stage of development in which horn flies arrest during pupal diapause. A study was therefore conducted to describe the morphologies of horn fly pupae and its central nervous system (CNS) throughout nondiapause pupal development and diapause. Morphologies of diapausing pupae and CNS indicated that developmental arrest occured early in pupal development during the interval between head eversion and pupal-adult apolysis. Morphological descriptions are necessary for defining compariable tissues between nondiapausing insects and diapausing insects. These tissues can then be used for molecular differential analysis to determine genes specific to either diapause or nondiapause. One such differential analysis technique, subtractive hybridization, was used to isolate putative diapause up-regulated genes from the horn fly. Seven different cDNAs were closed and partially sequenced. Comparisons of the cDNA sequences with known DNA and protein sequences indicated homology with transferrin, cytochrome oxidase I, Kunitz family serine protease inhibitor, tyrosine hydroxylase (TH), and carboxylensterase. Two cDNAs did not have homology to entries in DNA and protein databases. Northern blot analyses were used to study expression of each gene by probing total RNA extracted from whole pupae throughout nondiapause pupal development and diapause. Expression of TH was also determined in total RNA extracted from CNS tissue of nondiapausing and diapausing pupae. Cytochrome oxidase was equally expressed in nondiapause and diapause destined pupae, and therefore not considered to be a diapause up-regualted gene. Expression patterns differed slightly for each of the remaining clones; however, expression tended to be highest in diapause destined pupae during pupation compared to nondiapausing pupae. These genes and their products are involved in many aspects of insect phsiology including metamorphosis, melanization and sclerotization of the puparium and cellular defense. The possible functions of these genes and products are discussed in the context of the diapause process. / xii, 92 leaves : ill. ; 28 cm.

No description available.

Dormancy characteristics and optimum conditions for germination of Cleome gynandra seeds have not been explained. Seed storage proteins were extracted, analysed with SDS-PAGE and sequenced. Seed proteins of Cleome were characterised by comparison with those of wild mustard (Brassica kaber). Wild mustard showed seed proteins composed of two α-chains of molecular weight (24-32 kDa) and another two β-chains of 18-22 kDa. The seed proteins of Cleome comprised two α-chain polypeptides of molecular weight (25-30 kDa), two β-chain polypeptides of molecular weight (18-20 kDa) and a smaller β-chain of 13-15 kDa. The storage proteins occurred in the seeds as dimeric complexes of molecular weight 40-65 kDa, which were broken into polypeptide chains of approximately 20 and 30 kDa by the reducing . action of DTT. Comparison with proteins in the proteome library and similarity index further confirmed that the seed proteins of Cleome had similarities with those of wild mustard. Two dimensional SDS-PAGE showed that the two species have nine similar polypeptides and four different ones. Events associated with dormancy release during seed germination still require explanation. Seeds of Cleome are characterised by low germination and there has been no explanation for this. Changes in protein expression during germination of Cleome in the presence or absence of light and at constant or alternating temperatures were examined. The germination of Cleome seeds at 20 degrees C was inhibited by light, but it was improved at 20 degrees C in darkness. There was no photoinhibition when seeds were germinated at constant 30 degrees C or alternating 20/30 degrees C (16 h night and 8 h day) for 10 days. Four proteins were observed to decrease in expression as germination progressed, but remained unchanged during photoinhibition. Photoinhibition was expressed more in seeds that were harvested late, after the pods had turned brown. These seeds showed a fifth, low molecular weight protein (13 kDa) that was absent from the immature seeds and embryos. Photinhibition is a pseudo-dormancy condition during which seed storage proteins are not utilised and the seed coat could partially play a role in it. The temperatures for the germination of Cleome in darkness have been determined. However, prior to this study the effects of temperature, light and pre-germination treatments (chilling, scarification, hydration and germination in the presence of KN0(3) or GA(3) on the germination of the seeds of this species have not been investigated. Seeds were germinated for 10 days and the final count of germination was used to determine seed performance. The highest germination percentage (60% and 80%, for a 2-year old and a l-year old seed lot, respectively) of untreated seeds was achieved when alternating temperatures of 20/30 degrees C (16 h/S h) in the dark or constant 30 degrees C in the dark were used. Among the pre-germination treatments, only scarification (puncturing of seeds at the radicle end) improved germination. Seeds were found to be negatively photoblastic, and the phenomenon was more pronounced when they were germinated at 20 degrees C and 12 h photoperiod or longer. Germination of photoinhibited seeds was, however, improved by treatment with GA(3) It is recommended that the germination of Cleome be undertaken under conditions of darkness and at either alternating 20/30 degrees C or continuous 30 degrees C. Seed lot vigour and seedling vigour are two important seed quality aspects that are used in defining the seed germination process. Seed germination is appropriately characterised by radicle protrusion and the attainment of normal seedling structures. However, the international rules for testing seeds combine radicle protrusion and normal seedling attainment in separating seed germination into the first and final counts. The challenge to a seed analyst testing the germination of a species whose first and final counts are unknown is that there is no statistical guideline to determine these important stages of seed germination. Cauliflower and broccoli, for which the first and final counts are published in the international rules for testing seeds and Cleome, for which there is no data on the first and final counts, were examined to determine the statistical significances of the first and final counts. Analysis of variance, logistic regression, 'broken-stick' regression models and survival analysis procedures were used. Analysis of variance showed that there were no differences between the germination percentages on the fourth, fifth and seventh days of germination. Low and stable standard deviations were recorded when evaluating germination after the fourth day. The germination curves of broccoli and cauliflower did not fit the Gompertz curve but fitted the exponential curve. The broken-stick model 'broke' the cumulative germination curve for the Cleome seed lots into two linear curves that were significantly different, but failed to break those for broccoli and cauliflower. However, this study confirmed the first and final counts for broccoli and cauliflower as determined by the international rules for testing seeds. Broken-stick modelling and life table analyses confirmed the fourth day as being appropriate to determine the first count for Cleome germination. There was no evidence of further seed germination after the seventh day as shown by probability density and hazard rate. It is suggested that for Cleome, the 'first count' and 'final count' be performed on the fourth and seventh day of the germination, respectively. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

Preharvest sprouting (PHS), the germination of seeds on the maternal plant before harvest, is a big challenge for barley producers worldwide. It is attributed mainly to low seed dormancy. The balance between two classical plant hormones, gibberellin (GA) and abscisic acid (ABA) regulates seed dormancy and germination, and the endogenous level of these two hormones in plants is determined by their biosynthesis and catabolism. This thesis characterized the expression patterns of the major GA and ABA metabolism genes in barley cv. Betzes during seed development, and germination in both dormant and non-dormant seeds. The results indicate that specific gene family members of the two hormones play distinct temporal roles in regulating seed development, dormancy onset and release, and germination. Since only two genes encoding the GA deactivating GA 2-oxidase enzyme have been known so far in barley, this study also identified two new GA2ox genes designated as HvGA2ox1 and HvGA2ox3.

The Panama Canal Watershed (PCW) represents a special opportunity for studies related to seed dormancy in the tropics with both applied and basic research objectives. There is a clear need for seed information for nursery planning in restoration projects that involves use of a large number of local species. Moreover, the strong rainfall gradient along the 60 km of continuous lowland forest in the PCW represents an excellent opportunity to understand dormancy and its role as an adaptive trait that evolved in response to environmental factors. This study presents useful seed information for 100 tree species native to the PCW. For each species, it includes collection system, fruiting time, seed mass, seed moisture content, germination, and longevity in storage at 20??C. For the first time, an attempt is made to classify (to class sensu Baskin andamp; Baskin 2004) the class of dormancy in seeds of the PCW. An analysis of the relationship between class of dormancy and seed mass, moisture content, longevity, germination patterns, and seasonality is presented. The relationship between class of seed dormancy and longevity and geographical distribution of species within the watershed based on the rainfall gradient was investigated.

Inconsistent seed germination poses a problem for efficient seedling production of Echinacea species. Evidence suggests that ethylene can be effective for improving germination in Echinacea species. The objectives of this research were: to develop an ethylene pre-germination treatment that enhances germination in Echinacea species that is retained following drying and storage, and to determine if the ethylene effect on enhanced germination was an important mode of action for dormancy release. Four species of Echinacea (E. purpurea, E. tennesseensis, E. angustifolia and E. simulata) treated with 1-aminocyclopropane-1-carboxylic acid (ACC) or ethephon resulted in faster and generally higher germination. Pre-treatment of seeds with ACC or ethephon followed by drying was as effective as chilling stratification for enhancing germination depending on the species. While ethylene pretreatments did increase germination to some extent depending on species, it was concluded that 60-day stratification alone was a more commercially-viable treatment. Ethylene production or perception was not necessary for germination in untreated or stratified seeds as shown by aminoethoxyvinylglycine (AVG), silver thiosulfate (STS), and 1-methylcyclopropene (MCP) treatments. Both stratification and ACC treatment reduced Echinacea seed sensitivity to ABA and could be a common mechanism for enhanced germination. However, it does not appear that the increased germination seen after stratification was mediated through ethylene production because final germination percentages were generally unchanged following inhibition of ethylene production or action. In contrast, inhibition of ethylene production and perception reduced early 3-day germination suggesting that ethylene was more involved in seed vigor than final germination. It was determined that there is no physiological significance of ethylene for dormancy release in these Echinacea species.

Preharvest sprouting (PHS), the germination of seeds on the maternal plant before harvest, is a big challenge for barley producers worldwide. It is attributed mainly to low seed dormancy. The balance between two classical plant hormones, gibberellin (GA) and abscisic acid (ABA) regulates seed dormancy and germination, and the endogenous level of these two hormones in plants is determined by their biosynthesis and catabolism. This thesis characterized the expression patterns of the major GA and ABA metabolism genes in barley cv. Betzes during seed development, and germination in both dormant and non-dormant seeds. The results indicate that specific gene family members of the two hormones play distinct temporal roles in regulating seed development, dormancy onset and release, and germination. Since only two genes encoding the GA deactivating GA 2-oxidase enzyme have been known so far in barley, this study also identified two new GA2ox genes designated as HvGA2ox1 and HvGA2ox3.

The sucrose-nonfermenting-1 related kinase 1 (SnRK1) protein complex is a heterotrimeric serine/threonine protein kinase complex conserved in eukaryotes that acts as a regulator of carbon metabolism and energy homeostasis. The objective of this study was to determine if the SnRK1 protein complex has a role in the nitrogen response and during dormancy acquisition in poplar. Gene expression profiling of the PtdSnRK1, PtdAKINbeta, and PtdAKINgamma gene family members was carried out using a robust qRT-PCR assay. A subset of these genes showed modified expression patterns under differential nitrogen availability and during dormancy acquisition, suggesting that SnRK1 complexes comprised of specific subunits may be involved in the regulation of the response to nitrogen and during dormancy acquisition. The regulatory subunits PtdAKINbeta1.1, PtdAKINgamma1.1 and PtdAKINgamma2.3 were often identified using principal component analysis as significantly responsible for distinguishing treatments from one another and therefore merit further study. / Plant Biology

No description available.