Subterranean clover mottle virus (SCMoV) is a positive sense, single-stranded RNA virus that infects subterranean clover (Trifolium subterraneum) and a number of related legume species. The ultimate aim of this research was to investigate aspects of SCMoV that would support its use as a gene silencing vector for legume species, since RNA (gene) silencing is now a potential tool for studylng gene function. The ability of viruses to induce an antiviral defense system is being explored by virus-induced gene silencing (VIGS), in which engmeered viral genomes are used as vectors to introduce genes or gene ii-agments to understand the function of endogenous genes by silencing them. To develop a gene silencing vector, a number of aspects of SCMoV host range and molecular biology needed to be studied. A requirement for a useful viral vector is a suitably wide host range. Hence the first part of this work involved study of the host range and symptom development of SCMoV in a range of leguminous and non-leguminous plants. The aim of this work was to identify new and most suitable hosts among economically important crop and model legumes for functional genomic studies, and also to study symptom development in these hosts for comparison with host responses to any SCMoV-based viral vectors that might be used in later infection studies. A total of 61 plant genotypes representing 52 species from 25 different genera belonging to 7 families were examined for their response to SCMoV infection, including established and new crop legumes, established pasture, and novel pasture and forage legumes, and 12 host indicator plants belonging to the families Amaranthaceae, Apiaceae, Chenopodiaceae, Cruciferae, Cucurbitaceae and Solanaceae. Following mechanical inoculation, plants were examined for symptoms and tested for primary and secondary infection by RT-PCR andlor ELISA after 2-3 weeks and 3-9 weeks, respectively. Thirty-six legume hosts belonging to eight different genera of legumes were identified as suitable hosts of SCMoV, 22 of them systemic hosts and 15 were infected locally. Only two non-legumes were infected with SCMoV-P23, one systemically and one as a local host, so confirming that SCMoV is essentially a legume-infecting virus. This work considerably expanded knowledge of the host range of SCMoV. To provide the information needed to modify the SCMoV genome to develop gene vectors, the virus was characterized in detail. The complete genomes of four isolates, SCMoV-AL, SCMoV-MB, SCMoV-MJ and SCMoV-pFL, were sequenced using high fidelity RT-PCR and molecular cloning, and compared to the first sequenced isolate (SCMoV-P23) to give a complete picture of the genome organisation of the virus. The 4,258 nucleotide (nt) sequence of SCMoV RNA is not polyadenylated. The 5' non-coding region (NCR) is 68 nt in length and the 3' NCR is 174 nt. The coding regon contains four overlapping open reading fi-ames (ORFs). The first, OW1 (nt 68-608), encodes a putative protein containing 179 amino acids with a calculated molecular mass (Ma,) of 20.3 kDa. It overlaps with the next ORF, ORF2a, by four bases. ORF2a (nt 605-2347) encodes a putative protein of 580 amino acids with a Ma, of 63.7 kDa and contains a motif characteristic of chymotrypsin-like serine proteases. The ORF2b is probably translated as part of a polyprotein by -1 ribosomal fiameshifting in ORF2a. The transfiame product (Ma, = 107.5 kDa) is made up of 966 amino acids. A GDD motif typical of RNA virus polymerases is present in ORF2b. The 3' terminal ORF3 (nt 3323-4084) encodes the 27.3 kDa coat protein (CP). Nucleotide variation between the complete sequences of the isolates was two to three orders of magnitude larger than base misincoporation rates of the polymerases used in RT-PCR. Molecular relationship analysis between all five isolates, undertaken with the complete nucleotide sequences, clearly separated them into three groups. These groups reflect similar significantly diverse groupings based on the symptoms and their severity in subterranean clover. Intra-isolate sequence variability is therefore a possible cause of the differences in symptom severity. The analysis also showed that there were more nucleotide substitutions at the 5' terminal half of SCMoV than at the 3' end. This observation was confirmed by the higher value of nucleotide diversities at nonsynonymous versus synonymous sites (dN/ds ratio) estimated for the ORF1, compared to the near conservation of sequences of the other ORFs. These results, together with functional and comparative sequence analysis with other sobemoviruses, implicate the ORFl gene product in pathogenicity of SCMoV, possibly as a severity determinant or as a viral suppressor of RNA silencing in plants. Because more information on SCMoV genome function was required, the possible involvement of the ORFl gene product (PI) and the CP in movement of SCMoV was studied in cells of grasspea (Lathyrus clymenum L) and chickpea as C-terminal fusion constructs with jellyfish (Aequorea victoriae) green fluorescent protein (GFP). A transient expression vector, pTEV, for in planta synthesis of reporter gene constructs was developed. The vector was based on pGEM-T with 35s RNA transcriptional promoter of Caulzjlower Mosaic virus (CaMV) and nopaline synthase gene transcription terminator signal (T-Nos) separated by a multiple subcloning site. A custom-made particle inflow gun was used to introduce the constructs into plant cells. The bombardment conditions were fxst optimised for efficient delivery of DNAcoated particles. Transient gene expression of GFP was monitored 24-96 hours after particle bombardment. Fluorescence from GFP alone, GFP:CP and GFP:Pl constructs was observed in the nucleus of single cells, cytoplasm and cell periphery of neighbouring cells. There was limited spread of these fusion proteins from one cell to another 36-48 hours after transformation. These results indicate that the P 1 and CP cannot move independently from cell to cell. Other viral/cellular components might be needed to form a complex with these proteins to transport the viral genome. Putative nuclear export signals in the P1 and CP sequences of SCMoV were identified by sequence comparison. These could be tested by mutagenesis using full-length infectious clones. To determine the possibility of gene expression of vectors based on SCMoV, three forms of a full-length cDNA clone of SCMoV-pFL were developed: one with no heterologous transcriptional factors (pFL), a second under the control of only 35s (p35SFL) and a third with 35s and T-Nos (pTEVFL). Fifteen day-old in vitro-cultured chickpea, grasspea and subterranean clover seedlings were inoculated by particle bombardment using gold particles coated with plasmid pTEVFL. In vivo-transcribed RNA transcripts were detected by RT-PCR after two weeks in grasspea but not in subterranean clover and chickpea. Experiments were undertaken towards developing the SCMoV genome into a VIGS vector. Three forms each of five major GFP chimeric constructs of pFL (the full length SCMoV cDNA clone) were generated from which in vitro- and in vivo-transcribed RNA transcripts could be derived. The rationale used in developing these constructs was gene insertion andlor replacement with d p , and duplication of the putative subgenomic RNA promoter (sgPro) of SCMoV. The major constructs were as follows: pFLCPgfp, pFL with the d p gene fused to the 3' end of the CP gene, pFLP 1 gfp, pFL with gj27 gene fused to the 3 ' end of the ORF 1, pFLCPsgprogfp, pFL with a putative sgPro sequence and a translatable & gene cloned in tandem between the CP gene and the 3' NCR of SCMoV, pFLCPVsgprogf$, pFL with a putative sgPro sequence and a translatable gfp gene cloned in tandem between a truncated CP gene and the 3' NCR and pFLREPsgprog@, pFL with the ORF2b, a putative sgPro sequence and a translatable &fP gene cloned in tandem between a truncated CP gene and the 3' NCR These constructs were all made, but a detailed assessment of their vector potential could not be done because there was a delay of about one year whilst the Office of the Gene Technology Regulator processed the application for permission for glasshouse testing. Although additional work needs to be undertaken to complete development of a final RNA silencing vector, this study has contributed to new knowledge in terms of extending understanding of SCMoV host range, symptoms, sequence variation and control of gene expression. The constructs made have also laid the groundwork for development of a legume gene silencing vector based on SCMoV.

Thesis (Ph. D.)--University of Adelaide, 1984. / Some mounted ill. Includes bibliographical references (leaves 156-186).

Thesis (Ph.D.)--Murdoch University, 2005. / Thesis submitted to the Division of Science and Engineering. Bibliography: leaves 184-207.

Until recently, it was assumed that African mole-rats search for food randomly, as it is in concordance with Aridity food distribution hypothesis. However, recent studies indicate that some subterranean rodent species including mole-rats could be able to use plant chemicals (kairomones) for food localization. In my master thesis I conducted a battery of experiments on four mole-rat species to prove, that these species also posses the ability of kairomone guided foraging independently of their social organization.

Submitted by Laís Alves Maroubo null (lmaroubo@hotmail.com) on 2017-06-21T17:51:45Z No. of bitstreams: 1 20-06-17 - Dissertação de Mestrado_LaisMaroubo_para impressao_c ficha catalografica_CAPA DURA.pdf: 3730756 bytes, checksum: 2c57a3634736f0933b0e37a586f0a2a8 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-06-21T18:29:01Z (GMT) No. of bitstreams: 1 maroubo_la_me_sjrp.pdf: 3730756 bytes, checksum: 2c57a3634736f0933b0e37a586f0a2a8 (MD5) / Made available in DSpace on 2017-06-21T18:29:01Z (GMT). No. of bitstreams: 1 maroubo_la_me_sjrp.pdf: 3730756 bytes, checksum: 2c57a3634736f0933b0e37a586f0a2a8 (MD5) Previous issue date: 2017-05-05 / Os diagnósticos ambientais que abrangem as águas subterrâneas são importantes para manutenção e conservação da qualidade das águas dos aquíferos. Assim, identificar e quantificar os elementos potencialmente tóxicos presentes nas águas de poços corrobora para o desenvolvimento de ações mitigadoras do impacto ambiental a fim de prevenir que determinada contaminação evolua. A região de Presidente Prudente/SP é banhada pelo aquífero Bauru, que tem a água subterrânea captada para fins domésticos, agrícolas e industriais como fonte alternativa para o consumo. Deste modo, o principal objetivo deste trabalho foi a determinação e quantificação de íons metálicos potencialmente tóxicos, sendo estes, o cádmio (Cd2+), chumbo (Pb2+) e cobre (Cu2+) em amostras de água subterrânea, a fim de realizar um diagnóstico ambiental na área urbana de Presidente Prudente/SP. A determinação e quantificação dos metais Cd2+, Pb2+ e Cu2+ em amostras de água subterrânea foi realizada pela técnica voltamétrica de pulso diferencial por redissolução anódica utilizando como eletrodo de trabalho o eletrodo de mercúrio (Hanging Mercury Droping Electrode - HMDE), que se mostrou eficiente e satisfatória, comprovado pelos baixos limites de detecção dos metais. Para cada amostra, foram efetuadas medidas de pH, nas quais foram identificados o caráter básico em regiões situadas na FCT-Unesp e arredores, mantendo sua neutralidade em regiões mais distantes e menos populosa. Por meio de análises estatísticas, analisaram-se as características dos poços em função das profundidades dos poços artesianos, valores de pH e concentração dos metais. Os resultados das análises estatísticas pelo método das análises dos componentes principais (PCA) foram avaliados e verificou-se um comportamento semelhante e excedente para Cd e Pb em poços mais rasos, e independente e regular para Cu dentre os demais poços. Por fim, observou-se a relação de proporcionalidade entre profundidade e os metais Cd e Pb e a similaridade entre as medidas de pH entre dois poços próximos entre sí. / Environmental diagnostics covering groundwater is important for maintenance and conservation of water supplies quality. Thus, identifying and quantifying potentially toxic elements in the well water supports for the development of environmental impact mitigation actions to prevent contamination that particular evolve. The region of Presidente Prudente/SP is bathed in Bauru aquifer, which has groundwater abstracted for domestic, industrial and agricultural production as an alternative source for consumption. Thus, the main objective of this work was the determination and quantification of potentially toxic metal ions, which are, cadmium (Cd2+), lead (Pb2+) and copper (Cu2+) in groundwater samples in order to carry out an environmental diagnosis in the urban area of Presidente Prudente/SP. The determination and quantification of Cd2+ metals, Pb2+ and Cu2+ in groundwater samples was performed by technique anodic stripping voltammetric with mercury electrode (Hanging Mercury Droping Electrode - HMDE), which has proved efficient and satisfactory as evidenced by the low limits of detection of metals. For each sample, pH measurements were taken, where it was identified the basic character in regions situated in the FCT-Unesp and surroundings, maintaining its neutrality in more remote and less populated regions. Through statistical analysis, we analyzed the characteristics of the wells as a function of depths of the wells, pH and concentration of the metals. The results of statistical analysis by the method of principal components analysis (PCA) were evaluated and found a similar behavior to and exceeding Cd and Pb in shallower wells, and Cu regular independent from the other wells. Finally, there is the proportionality relationship between depth and Cd and Pb and similarity between pH measurements in two wells next to each other.

Orientador: Ana Maria Costa Leonardo / Banca: José Chaud Netto / Banca: Maria Santina de Castro Morini / Resumo: O forrageamento ou busca por alimento nos cupins subterrâneos envolve os operários, os soldados e em menor número as ninfas. O presente estudo foi realizado visando esclarecer aspectos da biologia do forrageamento de ninfas, o qual é pouco conhecido na espécie exótica Coptotermes gestroi. O monitoramento por meio de coletas periódicas em um período de 22 meses em 7 diferentes colônias mostrou que as ninfas forrageiras são produzidas durante todos os meses do ano. Provavelmente, devido à influência tanto de fatores externos como internos, cada colônia de Coptotermes gestroi possui uma dinâmica diferente de formação desses indivíduos. O estudo da biometria das ninfas forrageiras foi realizado para determinar os ínstares presentes nas colônias, sendo que as variáveis analisadas foram largura da cabeça, largura do pronoto, comprimento do broto alar, comprimento da tíbia, região de crescimento da antena e comprimento do corpo. Entretanto, as variáveis região de crescimento da antena e comprimento do corpo, inicialmente propostas, não se mostraram viáveis para a discriminação dos ínstares. Os dados foram submetidos à Análise dos Componentes Principais (ACP) e plotados em diagramas de dispersão para a discriminação dos ínstares. Das 533 ninfas forrageiras coletadas, 34 eram de 3° instar, 280 eram de 4° instar e 219 eram de 5° instar. Apesar de terem sido coletadas ninfas de 3° instar, provavelmente, estas não participam das atividades de forrageamento uma vez que foram coletadas em apenas uma colônia e em quantidade ínfima. O 5° instar ninfal foi coletado nas 8 colônias estudadas, contudo o 4° instar ninfal foi mais numeroso, mesmo tendo sido coletado em apenas 4 das colônias... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The foraging or search for food in subterranean termites involves workers, soldiers and in a small number nymphs. The present study was carried out aimed at clarifying aspects of the nymph foraging biology, which is little known in the exotic species Coptotermes gestroi. The monitoring through regular collections in 7 different colonies for a period of 22 months showed a production of forager nymphs all year round. Probably due to the influence of both internal and external factors each colony of Coptotermes gestroi has a different formation dynamic of these individuals. The biometric study of the forager nymphs was conducted to determine the instars present in the colonies, and the variables head width, pronotum width, wing bud length, right hind tibia length, antennal growing region and body length were measured. However, the variables antennal growing region and body length, originally proposed, were not feasible to instars discrimination. The data were submitted to the Principal Component Analysis (PCA) and plotted on a scatter diagram to determine the instars. Of the 533 forager nymphs collected, 34 were from 3rd - instar, 280 were from 4th - instar and 219 were from 5th - instar. Although 3rd - instar nymphs have been collected, probably, they do not participate in the foraging activities since they were collected only in one colony and in small quantity. The 5th - nymphal instar was collected in 8 colonies, however the 4th - nymphal instar was more numerous, even being collected in only 4 of all colonies. In order to characterize the instars, the eye color and the number of antennal segments in the of forager nymphs were recorded. These individuals may have light brown or white eyes, the nymphs with white eyes appeared in larger number and were present in all colonies. The sex determination of male and female nymphs was conducted... (Complete abstract click electronic access below) / Mestre

Sight in subterranean mammals living in a dark ecotope has generally been assumed as not needed and therefore greatly diminished in its function. Recent neuroanatomical studies demonstrate unexpected preservation of the visual system of several African mole-rats (Bathyergidae, Rodentia). Only a few behavioural studies, testing visual abilities and discussing their adaptive significance in these rodents, have been published to date. A spontaneous preference to light stimuli of two mole-rat species, the silvery mole-rat (Heliophobius argenteocinereus) and the giant mole-rat (Fukomys mechowii), was tested in this study. Assessed results showed convincingly that both species are able to perceive light. The following experiments provided the first behavioural support to the perception of short-wavelengths in this intensively studied group of subterranean rodents.

Um dos principais objetivos da Biologia Subterrânea é identificar e entender as modificações morfológicas apresentadas pelos organismos subterrâneos que podem ser relacionadas a seu modo de vida. No presente trabalho, realizamos essa investigação utilizando como modelo as espécies subterrâneas de Ituglanis Costa & Bockmann, 1993 (Siluriformes: Trihomycteridae). Atualmente são conhecidas sete espécies subterrâneas do gênero, as quais se distribuem em regiões geográficas muito próximas entre si, as áreas cársticas de São Domingos e de Mambaí e Posse, no nordeste do estado de Goiás. Analisamos caracteres externos (tamanho e alonga-mento corporal, nadadeiras pares e pigmentação) e sensoriais (olhos, barbilhões, sistema láte-rosensorial e encéfalo), e identificamos tendências de modificação em todos eles. As espécies subterrâneas de Ituglanis tendem a apresentar: alongamento do corpo e, nas espécies que tran-sitam ou ocupam espaços confinados, redução do tamanho corporal; nadadeiras peitorais mais longas e com mais raios, e as pélvicas reduzidas; pigmentação reduzida, os cromatóforos me-nos abundantes e com formas irregulares; olhos menores, deformados, assimétricos e encober-tos por pele; barbilhões bem desenvolvidos, os maxilares mais longos; sistema láterosensorial de canais fragmentado e reduzido, com grande variabilidade e assimetria; e o encéfalo com lobos vagais e faciais desenvolvidos, lobos ópticos reduzidos e aumento do tamanho do cere-belo, da bexiga natatória e dos canais semicirculares do ouvido interno. Muitas dessas caracte-rísticas já haviam sido observadas em espécies subterrâneas de outros grupos de peixes, suge-rindo que elas de fato estejam relacionadas a esse modo de vida. Nem todas as espécies sub-terrâneas de Ituglanis apresentam todas as características em um mesmo grau, de forma que é possível dividi-las entre as menos troglomórficas, Ituglanis sp. 1 e I. mambai, as intermediá-rias, Ituglanis sp. 2 e I. bambui, e as mais troglomórficas, I. passensis, I. ramiroi e I. epikars-ticus. A grande variabilidade tanto intra- quanto interespecífica sugere que essas característi-cas evoluem em mosaico, corroborando hipóteses anteriores de colonização independente do ambiente subterrâneo por cada uma das espécies. As modificações identificadas podem ser divididas em progressivas e regressivas, e discutimos os mecanismos evolutivos que explicam seu surgimento nas espécies subterrâneas. A evolução de características que conferem vanta- gem evolutiva é explicada por mecanismos de seleção natural direta, enquanto que os caracte-res regressivos que não apresentam vantagem ou desvantagem óbvia se encaixam em cenários de evolução neutra. Algumas das características das espécies subterrâneas são típicas de indi-víduos em estágios iniciais de desenvolvimento, o que sugere que a heterocronia pode ter um papel importante na evolução do troglomorfismo. Além disso, a linhagem a que pertencem as espécies subterrâneas apresenta características consideradas preaptativas para o modo de vida subterrâneo, o que favoreceria o desenvolvimento desse modo de vida repetidas vezes na fa-mília Trichomycteridae e, mais especificamente, no gênero Ituglanis. / One of the main objectives of Subterranean Biology is to identify and understand the morpho-logical modifications exhibited by subterranean organisms which can be related to their hypogean life habit. In the present work, we performed this investigation using as model the subterranean species of Ituglanis Costa & Bockmann, 1993 (Siluriformes: Trihomycteridae). Nowadays, seven subterranean species of the genus are recognized, distributed in close geo-graphic regions, the São Domingos karstic area and the Mambaí and Posse karstic area, both on northeastern Goias state, Brazil. We analyzed external (body size and elongation, paired fins and pigmentation) and sensorial characters (eyes, barbels, laterosensory system and brain), and identified tendencies of modification in all of them. Subterranean species of Ituglanis tend to shown: body elongation, and those species occupying confined spaces, body-size reduction; longer pectoral fins, with increased number of rays, and pelvic fins reduced; reduced pigmentation, the cromatophores less numerous and with irregular forms; smaller, deformed and asymmetric eyes, covered by skin; well developed barbels, the maxillary long-er; the canals of the laterosensory system fragmented and reduced, with variability and asymmetry; and the brain with well developed vagal and facial lobes, reduced optic lobes and increased cerebellum, swim bladder and semicircular canals of the inner ear. Many of these characters were already reported for subterranean fishes of other groups, suggesting a rela-tionship with this life habit. The subterranean species of Ituglanis does not exhibit all the characters on the same degree, in such a way that it is possible to classify them as the less troglomorphic, Ituglanis sp. 1 and I. mambai, the intermediary, Ituglanis sp. 2 and I. mambai, and the most troglomorphic, I. passensis, I. ramiroi and I. epikarsticus. The large inter- and intraspecific variability observed suggests a mosaic of evolution for these characters, corrobo-rating previous hypothesis of independent colonization of the subterranean environment in each species. The modifications identified can be divided in progressive and regressive, and we discuss the evolutionary mechanisms that explain their origin. The evolution of characters that can be evolutively advantageous is explained by mechanisms of direct natural selection, whilst regressive characters that do not provide any obvious advantage os disadvantage fit to scenarios of neutral evolution. Some of the characters observed in the subterranean species are typical of individuals in early stages of development, suggesting that heterocrony can play an important role in the evolution of troglomorphism. Furthermore, the lineage to which the subterranean species belong exhibit some characters considered preadaptative for the subter-ranean life habit, favouring its development repeatedly in the family Trichomycteridae and, more specifically, in the genus Ituglanis.

No description available.

Reticulitermids were significantly more likely to discover subterranean baits connected by physical guidelines than freestanding baits under both laboratory and field conditions. In the laboratory, subterranean termites built significantly longer tunnels adjacent to cellulosic guidelines than plastic guidelines. In the field, all guideline materials were equally effective at directing tunneling activity. Reticulitermes spp. workers were tested to determine their preferred substrate temperature. The preferred range for Reticulitermes spp. workers was found to be 18 to 27 degrees C. A laboratory bioassay was performed to determine if Reticulitermes spp. aggregates within thermal shadows. Significantly more Reticulitermes spp. workers aggregated within cool thermal shadows than control areas. In a multiple choice bioassay, mean consumption was higher for paper baits treated with fructose, galactose, glucose, raffinose, sucrose, trehalose and uric acid than for control baits. In a multiple choice bioassay, mean consumption was significantly lower for baits treated with arbutin, and most amino acids than for control baits. In the no-choice bioassay, the amount of paper bait consumed did not differ significantly for any of the treated baits tested and control baits. / Ph. D.