In this study adaptations of bryophytes and lichens to desiccation stress were examined. The aim was to test whether desiccation tolerance in the selected species is constitutive or if desiccation tolerance could be induced by various hardening treatments. In addition, some putative tolerance mechanisms were investigated, including the accumulation of sugars, increase in ROS scavenging systems and other mechanisms e.g. energy dissipating processes. To determine if hardening treatments prior to desiccation stress increased desiccation tolerance, mosses and lichens were partially desiccated or treated with ABA. The effect of hardening treatments on the physiology of the moss Atrichum androgynum and lichens Peltigera polydactyla, Ramalina celastri and Telochistes capensis during a desiccation-rehydration cycle was investigated. Photosynthesis, respiration and chlorophyll fluorescence measurements were used as rapid tools to determine the metabolic activities in these lichens and moss species. In A. androgynum partial desiccation following slow drying at 52% RH increased the rate of recovery of net photosynthesis. Net photosynthesis recovered almost completely following slow drying in the material that was partially dehydrated and/or treated with ABA. This suggests that partial dehydration hardens the moss, and that ABA can fully substitute for partial dehydration. In R celastri and P. polydactyla both partial dehydration and ABA treatments displayed some improvement in desiccation tolerance depending on the duration and severity of stress. The reduction in the re-saturation respiration burst in P. polydactyla, although not quite significant, strongly suggests that hardening increases mycobiont tolerance. However, it is more difficult to establish whether the hardening treatments improve photobiont performance. In the moss A. androgynum ABA treatment increased the rate of recovery of photosynthesis and PSII activity, and also doubled non-photochemical quenching (NPQ). Increased NPQ activity will reduce ROS formation, and may explain in part how ABA hardens the moss to desiccation. In ABA treated, but not untreated mosses, desiccation significantly increased the concentration of soluble sugars in A. androgynum. Sugar accumulation may promote vitrification of the cytoplasm and protect membranes during desiccation. Starch concentrations in freshly collected A. androgynum and R. celastri were only c. 40 and 80 mg g ¯¹ dry mass respectively, and slightly rose during desiccation, but were only slightly affected by ABA pretreatment. ABA did not reduce chlorophyll breakdown during desiccation. In P. polydactyla ABA pretreatment had little effect on any of these parameters. Changes in the activities of the free radical scavenging enzymes ascorbate peroxidase, catalase and superoxide dismutase were measured during wetting and drying cycles in the moss A. androgynum and in the lichens P. polydactyla, R. celastri and T capensis. These species normally grow in the understorey of the Afromontane forest, moist, xeric, and extremely xeric miicrohabitats respectively. In A. androgyum, enzyme activity was measured shortly after collection, after 3 d storage following hardening by partial dehydration and/or 1 h treatment with ABA or distilled water and during desiccation and rehydration. In A. androgynum enzyme activities of CAT and SOD in untreated material were always higher than in the hardened treatments, while both partial dehydration and ABA treatments tended to reduce both CAT and the induction of SOD activity, although these effects were not significant between the treatments. This suggests that ABA may not be involved in the induction of free radical scavenging enzymes and probably these enzymes are not important in desiccation tolerance of A. androgynum. In lichens, the enzyme activity was measured shortly after collection, after hydration for 48 hat 100% RH, after desiccation for 14 d and 28 d, and during the first 30 min of hydration with liquid water. Enzyme activities tended to rise or stay the same following rehydration in all the species tested. After desiccation for 14 d, enzyme activities decreased, and then decreased further to very low values after 4 weeks desiccation. In all species, including T capensis from an extremely xeric habitat, the activities of all enzymes remained at very low values during the 30 min following rehydration, and were therefore unavailable to remove any reactive oxygen species accumulating in lichen tissues as a result of desiccation. Results suggests that the enzymic antioxidants are more likely to be involved in removing reactive oxygen species produced during the normal metabolic processes of lichens than having a role in desiccation tolerance. The Afromontane understorey moss Atrichum androgynum displayed an oxidative burst of H₂O₂ during rehydration following desiccation. Maximum rates of H₂O₂ production occur during the first 15 min of rehydration. While the production of H₂O₂ increases with increasing desiccation times, the moss produced significant amounts of H₂O₂ during rehydration after desiccation for times that did not inhibit photosynthesis or cause K⁺ leakage. A. androgynum may produce more H₂O₂ during desiccation than rehydration, because desiccation artificially induced using polyethylene glycol strongly stimulates production. Experiments involving inhibitors and exogenously supplied reductants indicate that peroxidases are responsible for the synthesis of H₂O₂. Factors that influence the rate of H₂O₂ production during rehydration include light and the hormone ABA. Patterns of H₂O₂ production are discussed in terms of their possible role as a defence against pathogenic fungi and bacteria. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2002.

Orientador: Jairo Osvaldo Cazetta / Banca: Samira Domingues Carlin Cavallari / Banca: Cesar Martoreli da Silveira / Resumo: Pesquisas científicas focadas em identificar genótipos tolerantes a estresse abiótico são altamente desejáveis, pois, o uso desses genótipos diretamente no campo, ou indiretamente em processos de melhoramento genético, permite reduzir custos de manejo do solo, da cultura e perdas de produtividade. O objetivo desse trabalho foi determinar a tolerância de 24 genótipos de cana-de-açúcar sob elevados teores de Al3+ e Mn2+, associado a baixa disponibilidade de nutrientes. Para trabalhar com elevado número de unidades experimentais, um novo sistema de hidroponia foi adaptado e testado neste trabalho. O ensaio foi instalado e conduzido em casa de vegetação em delineamento inteiramente casualizado, num esquema fatorial 24x2, correspondendo a 24 genótipos, dois tratamentos (com e sem estresse), com quatro repetições, totalizando 192 unidades experimentais. No tratamento sem estresse, as plantas foram cultivadas em solução nutritiva completa e no tratamento com estresse foi utilizada a mesma solução nutritiva com elevada acidez (4,0 ± 0,1) e com apenas 5% da sua concentração original, além da elevada concentração de alumínio (60 mg L-1) e manganês (700 mg L-1) ambos na forma de cloreto. Os genótipos RB966928, RB855443, IACSP96-3060, SP81-3250, RB867515, CTC 21, RB965902 e IAC91-1099 foram os que tiveram suas características biométricas menos afetadas pelo estresse proposto e consequentemente foram considerados os mais tolerantes. Por outro lado, os genótipos RB965917, CTC 15, CTC17, RB... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The scientific research focused on identifying and understanding abiotic stress-tolerant genotypes is highly desirable since their use directly in the field or indirectly in breeding processes may reduce costs of soil and crop management and productivity losses. This aim of this study was to determine the tolerance of 24 sugarcane genotypes under high contents of Al3+ and Mn2+ associated with the low availability of nutrients. In order to work with a great number of experimental unities, a new way of hidroponic method was adapted and tested in this research. The experiment was carried out in a greenhouse in a completely randomized design, in a 24 × 2 factorial scheme, consisting of 24 genotypes, two treatments (with and without stress), and four replications, totaling 192 experimental units. In the treatments without stress, plants were grown in a complete nutrient solution whereas in the treatments with stress, a nutrient solution with a high acidity (4.0 ± 0.1) and only 5% of its original concentration, as well as a high concentration of aluminum (60 mg L-1 ) and manganese (700 mg L-1 ), was used, both in the form of chloride. The genotypes RB966928, RB855443, IACSP96-3060, SP81-3250, RB867515, CTC 21, RB965902 and IAC91- 1099 were those that had their biometric characteristics less affected by the proposed stress and consequently were considered the most tolerant. genotypes RB965917, CTC 15, CTC17, RB855536, CTC 2, CTC 20, and CTC99-1906 were identified as the most sensiti... (Complete abstract click electronic access below) / Mestre

Murine stress-inducible protein 1 (mSTI1) is a co-chaperone that is homologous with the human heat shock protein 70 (Hsp70)/heat shock protein 90 (Hsp90)-organizing protein (Hop). The two proteins are homologues of the highly conserved stress-inducible protein 1 (STI1) family of co-chaperones. The STI1 proteins interact directly and simultaneously at some stage, with Hsp70 and Hsp90 in the formation of the hetero-multi-chaperone complexes that facilitate the folding of signal transducing kinases and functional maturation of steroid hormone receptors. The interactions of mSTI1 with both Hsp70 and Hsp90 is mediated by a versatile structural protein-protein interaction motif, the tetratricopeptide repeat (TPR). The TPR motif is a degenerate 34-amino acid sequence a-helical structural motif found in a significant number of functionally unrelated proteins. This study was aimed at characterizing the structural and functional determinants in the TPR domains of mSTI1 responsible for binding to and discriminating between Hsp70 and Hsp90. Guided by data from Hop's crystal structures and amino acid sequence alignment analyses, various biochemical techniques were used to both qualitatively and quantitatively characterize the contacts necessary for the N-terminal TPR domain (TPR1) of mSTI1 to bind to the C-terminal EEVD motif of heat shock cognate protein 70 (Hsc70) and to discriminate between Hsc70 and Hsp90. Substitutions in the first TPR motif of Lys⁸ or Asn¹² did not affect binding of mSTI1 to Hsc70, while double substitution of these residues abrogated binding. A substitution in the second TPR motif of Asn⁴³ lowered but did not abrogate binding. Similarly, a deletion in the second TPR motif coupled with a substitution of Lys⁸ or Asn¹² reduced but did not abrogate binding. Steady state fluorescence and circular dichroism spectroscopies revealed that the double substitution of Lys⁸ and Asn¹² resulted in perturbations of inter-domain interactions in mSTl1. Together these results suggest that mSTI1-Hsc70 interaction requires a network of electrostatic interactions not only between charged residues in the TPR1 domain of mSTI1 and the EEVD motif of Hsc70, but also outside the TPR1 domain. It is proposed that the electrostatic interactions in the first TPR motif collectively made by Lys⁸ and Asn¹² define part of the minimum interactions required for successful mSTI1-Hsc70 interaction. In the first central TPR domain (TPR1A), single substitution of Lys³°¹ was sufficient to abrogate the mSTI1-Hsp90 interaction. Using a truncated derivative of mSTI1 incapable of binding to Hsp90, residues predicted by crystallographic data to determine Hsp70 binding specificity were substituted in the TPR1 domain. The modified protein had reduced binding to Hsc70, but showed significant binding capacity for Hsp90. In contrast, topologically equivalent substitutions on a truncated derivative of mSTI1 incapable of binding to Hsc70 did not confer Hsc70 specificity on the TPR2A domain. These data suggest that binding of Hsc70 to the TPR1 domain is more specific than binding of Hsp90 to the TPR2A domain. In addition, residues C-terminal of helix A in the second TPR motif of mSTI1 were shown to be important in determining specific binding to Hsc70. Binding assays using surface plasmon resonance spectroscopy showed that the affinities of binding of mSTI1 to Hsc70 and Hsp90 were 2 μM and 1.5 μM respectively. Preliminary in vivo studies revealed differences in the dynamics of binding of endogenous and exogenous recombinant mSTI1 with Hsc70 and Hsp90. The outcome of this study poses serious implications for the mechanisms of mSTI1 interactions with Hsc70 and Hsp90 in the cell.

Many stress situations limit plant growth, resulting in crop production difficulties. Population growth, limited availability and over-utilization of arable land, and intolerant crop species have resulted in tremendous strain being placed on agriculturalists to produce enough to sustain the world's population. An understanding of the principles involved in plant resistance to environmental stress will enable scientists to harness these mechanisms to create stress-tolerant crop species, thus increasing crop production, and enabling the farming of previously unproductive land. This research project uses computational and bioinformatics techniques to explore the promoter regions of genes, encoding proteins that are up- or down-regulated in response to specific abiotic stresses, with the aim of identifying common patterns in the cis-elements governing the regulation of these abiotic stress responsive genes. An initial dataset of fifty known genes encoding for proteins reported to be up- or down-regulated in response to plant stresses that result in water-deficit at the cellular level viz. drought, low temperature, and salinity, were identified, and a postgreSQL database created to store relevant information pertaining to these genes and the proteins encoded by them. The genomic DNA was obtained where possible, and the promoter and intron regions identified. The Neural Network Promoter Prediction (NNPP) software package was used to predict the transcription start signal (TSS) and the promoter searching software tool, TESS (Transcription Element Search Software) used to identify known and user-defined cis-elements within the promoter regions of these genes. Currently available promoter prediction software analysis tools are reported to predict one promoter per kilobase of DNA, whilst functional promoters are thought to only occur one in 30-40 kilobases, which indicates that a large perccntage of predictions are likely to be false positives (pedersen et. al., 1999). NNPP was chosen as it was rated as the highest performing promoter prediction software tool by Fickett and Hatzigeorgiou (1997) in a thorough review of eukaryotic promoter prediction algorithms, however results were less than promising as very few predicted TSS were identified in the area 50 bps up- and downstream of the gene start site, where biologically functional TSSs are known to occur (Reese, 2000; Fickett and Hatzigeorgiou, 1997). TESS results seemed to support the hypothesis that drought, low-temperature and high salinity plant stress response proteins have similar as-elements in their promoter regions, and suggested links to various other gene regulation mechanisms viz. gibberellin-, light-, auxin- and development-regulated gene expression, highlighting the vast complexity of plant stress response processes. Although far from conclusive, results provide a valuable basis for future comparative promoter studies that will attempt to deduce possible common transcriptional initiation of abiotic stress response genes. / KMBT_363 / Adobe Acrobat 9.54 Paper Capture Plug-in

Appreciation of the ecological and economic values associated with healthy salt marshes has led to a recent rise in the number of marshes that are being targeted for restoration by dike removal. The success of restoration is often measured by the return of marsh plants, though this overlooks a key component of salt marshes, that of the invertebrate community within marsh sediments. To evaluate the short-term recovery of these invertebrates, sediment cores were collected across an elevational gradient in a recent dike removal marsh, one and two years post removal, and a nearby reference marsh. Abundance, richness and diversity as well as morphospecies community composition were compared across treatment groups (Reference, Removal) and elevation zone (High Marsh, Low Marsh). Morphospecies richness, abundance and diversity were significantly higher in Low Marsh samples than in High Marsh samples, though no statistically significant differences were found across treatments of the same elevation (e.g., Reference Low Marsh versus Removal Low Marsh). Pair-wise ANOSIM results found significant differences between community compositions across treatments, specifically Reference Low Marsh and Removal Low Marsh. The marsh edge, the lowest point of vascular plant growth before transitioning to tide flats, is considered a high stress environment for emergent vegetation. Plant establishment and survival in this low elevation zone is limited by the tolerance to inundation duration and frequency and anoxic sediments. Bioturbation and burrowing by macroinvertebrates increases the surface area exposed to surface water for gas exchange, increasing the depth of the redox potential discontinuity layer. Crabs that make stable, maintained burrows have been shown to increase oxygen penetration into sediment, improving plant productivity. Such crabs are not found in salt marshes of the Pacific Northwest of North America. However, other burrowing invertebrates may have a positive impact on plant health in these areas by reducing abiotic stress due to anoxic sediments, thereby allowing plants to establish and survive lower in the intertidal zone. To assess this potential relationship, study plots of Distichlis spicata were selected at equivalent elevations at the lowest point of plant establishment at the marsh edge. Focal plant rhizomes were severed from upland ramets and assigned an invertebrate abundance treatment based on a visual burrow count surrounding each plant (9 cm diameter). Focal plants were visited monthly from July to September 2016, plant health variables of chlorophyll content and chlorophyll fluorescence (photosynthetic efficiency), and sediment ORP readings were collected. Plant survivorship was significantly higher in plots with invertebrates, 96% of plants in 'With Invertebrate' plots and 50% of plants in 'No Invertebrates' plots survived the duration of the study. Plant health (chlorophyll content and chlorophyll fluorescence) generally increased with increased invertebrate presence though, not statistically significant. There may be potential for improved plant productivity and resilience to plants at the marsh edge due to invertebrate burrowing activity. This benefit could help mitigate projected losses in plant productivity due to sea level rise, though more research is needed to investigate the mechanism by which these invertebrates confer a health benefit to plants at the marsh edge.

Significant alterations in the environmental conditions can have pronounced effects on plant genome stability. Recent evidence argues for a global involvement of the components of epigenetic modules in the regulation of genome homeostasis both immediately after stress exposure and long after environmental cues were acquired. The last observation is of particular interest as the memory of imposing stress can be maintained at the molecular level throughout plant ontogenesis and may be faithfully propagated into the following generation. Our study provides evidence that epigenetic repercussions exerted by stress exposure of parental plants manifest themselves in untreated progeny at all three levels of the epigenetic module: DNA methylation, histone posttranslational modifications and small RNA metabolism. Additionally, the results of our study shed new light on the engagement of the epigenetic machinery in the maintenance of plant genome integrity by counteracting the activity of invading nucleic acids. / xv, 280 leaves : ill. ; 29 cm

[No abstract available] / xi, 176 leaves : ill. (chiefly col.) ; 29 cm

Physiological responses of six sorghum (Sorghum bicolor (L.) Moench) hybrids and their respective parental lines were evaluated under high and low soil moisture conditions at Tucson, Arizona in 1980 and 1981. Apparent photosynthesis, transpiration, diffusive resistance, and temperature differential (ambient temperature minus leaf temperature) were measured under field conditions at weekly intervals. To measure apparent photosynthesis, a small plexiglas chamber was sealed over a section of leaf blade and gas was sampled with two syringes pulled at a 30 or 60 second interval. The gas samples were injected from the syringes into an infrared gas analyzer to measure CO(,2) concentrations. Transpiration, diffusive resistance, and temperature differential were measured with a steady state porometer. Regression analysis was used to compare the physiological performance of the germplasm sources over a wide range of environmental conditions. The physiological characteristics of the highest yielding sorghum hybrid in 1980 were the most stable across all environments. This hybrid was superior to both its male and female parent for all four physiological characteristics. In 1980, this hybrid was superior to the other hybrids in temperature differential and transpiration. The superior yielding hybrid had the highest mean apparent photosynthesis and the lowest mean diffusive resistance. All hybrids had higher yields than their respective male parents under both irrigation treatments. Thirty-five days after planting, the superior yielding hybrid had the greatest leaf, stem, and root dry weights. This hybrid also had the highest relative leaf area expansion rate. Heterosis for stomatal density was not observed either year. Regression analysis proved to be an effective tool for analyzing the sorghum germplasm over a diverse range of environmental conditions. By comparing the response of an individual germplasm against the mean response of the population under numerous environmental conditions, the slope, mean, and coefficient of correlation may be used to evaluate genotype-environment interactions.

The development, yield and water use of six sorghum (Sorghum bicolor (L.) Moench) hybrids and their respective male and female parents were evaluated under stressed and well irrigated conditions during 1980 and 1981 at Tucson, Arizona. Changes in soil moisture storage were measured by neutron modulation on a semiweekly schedule. Transpiration, diffusive resistance and leaf-ambient temperature differentials were monitored biweekly using a steady state porometer. Meteorological data was collected on a daily basis. The 1980 season had higher maximum temperatures and pan evaporation than the 1981 season. Differences in soil moisture extraction among sorghum entries were not apparent within water treatments. Mean cumulative evapotranspiration (ET) for the stressed treatment was 270 and 261 mm, for 1980 and 1981, respectively. Mean cumulative ET for irrigated treatment was about twice that at 520 and 648 mm during 1980 and 1981, respectively. There were no apparent differences in cumulative ET for entries in the stressed treatment while genotypic differences were manifested under well irrigated conditions. Temperature differential demonstrated a significant and negative correlation with diffusive resistance especially under stressed conditions (r = -.64 in 1981). Temperature differential was positively correlated with transpiration (r = .70 in 1980 stressed treatment). Plant height was significantly affected by water level both years while stem weight was affected by water level only in 1980. Soil moisture treatments did not affect leaf area either year and genotypic differences were demonstrated only in 1981. Hybrids produced greater grain yield than their male parents under both water treatments. This was due to greater seed number for hybrids. Seed number was also more stable for hybrids under both moisture levels. Hybrids four and seven had the greatest grain yield in 1980 and 1981, respectively. Harvest index was improved with increased water application due to increased seed number. Hybrid four in 1980 and hybrid seven in 1981 were extremely efficient in water use exhibiting ET ratios of 283 and 378, respectively under high water application. Reduced water application had little affect on the performance of these entries.

Orientador: Sonia Marli Zingaretti / Coorientador: Durvalina Maria Mathias dos Santos / Banca: Isabela Cristina Gomes Honório / Banca; Jackson Antonio Marcondes de Souza / Resumo: A cana-de-açúcar é atualmente uma das principais culturas da agroindústria mundial. Devido à ampla expansão de seu plantio é submetida constantemente a solos não produtivos. A presença de moléculas tóxicas no solo, como o alumínio (Al3+), interfere diretamente no desenvolvimento radicular ocasionando baixa absorção de água e nutrientes levando a pouca produtividade e desenvolvimento das plantas. Os microRNAs tem sido descritos como um dos fatores responsáveis pela regulação gênica e a descoberta dessas moléculas abre um novo caminho para a elucidação da tolerância e adaptação das plantas aos estresses abióticos. Este estudo visou avaliar em duas cultivares de cana-de-açúcar (CTC-2 e RB855453) a expressão dos microRNAs miR159, miR164 e miR168, associados à resposta ao alumínio em espécies como Arabidopsis thaliana, arroz (Oriza sativa) e tabaco (Nicotiana tabacum). Algumas características foram avaliadas, como densidade de raízes (DS), área foliar (AR), produção de massa seca (MS) e teor de prolina nas folhas, em quatro cultivares de cana-de-açúcar submetidas a diferentes concentrações de alumínio. A cultivar CTC 2 foi classificada como tolerante e a RB855453 como sensível ao estresse. Ambas foram selecionadas e em sistema de hidroponia submetidas novamente ao estresse pela toxidez de alumínio na concentração de 221 μmol L -1 . Parâmetros fisiológicos foram mensurados (área foliar, potencial osmótico, taxa de fotossíntese, transpiração, condutância estomát... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The sugarcane is currently one of the main crops of global agribusiness. Due to the wide expansion of its plantation is constantly subjected unproductive soils. The presence of toxic molecules in the soil, such as aluminum (Al3+), directly affects root development, leading to poor absorption of nutrients and water leading to low productivity and development of plants. Studies of the interactions of plants with the environment are being conducted to clarify the resistance or susceptibility of various cultures, favoring the discovery of important mechanisms that participate in physiological and molecular responses to environmental stresses. MicroRNAs have been described as one of the factors responsible for gene regulation and the discovery of these molecules opens a new path for the elucidation of tolerance and adaptation of plants to abiotic stresses. This study evaluated in two sugarcane varieties the expression of microRNA miR159, miR164 and miR168, associated with the response to the aluminum species such as Arabidopsis thaliana, rice (Oryza sativa) and tobacco (Nicotiana tabacum). Some characteristics were evaluated, such as density (DS), area (AR), dry mass production (DM) and proline content, in four sugarcane cultivars submitted to different concentrations of aluminum. CTC 2 cultivar was classified as tolerant and RB855453 as stress sensitive. Both were selected and in a hydroponics system again submitted to stress by the aluminum toxicity in the conc... (Complete abstract click electronic access below) / Mestre