The resurrection plant Sporobolus stapfianus: An unlikely model for engineering enhanced plant biomass?

The resurrection grass Sporobolus stapfianus Gandoger can rapidly recover from extended periods of time in the desiccated state (water potential equilibrated to 2% relative humidity) (Gaff and Ellis, Bothalia 11:305–308 1974; Gaff and Loveys, Transactions of the Malaysian Society of Plant Physiology 3:286–287 1993). Physiological studies have been conducted in S. stapfianus to investigate the responses utilised by these desiccation-tolerant plants to cope with severe water-deficit. In a number of instances, more recent gene expression analyses in S. stapfianus have shed light on the molecular and cellular mechanisms mediating these responses. S. stapfianus is a versatile research tool for investigating desiccation-tolerance in vegetative grass tissue, with several useful characteristics for differentiating desiccation-tolerance adaptive genes from the many dehydration-responsive genes present in plants. A number of genes orthologous to those isolated from dehydrating S. stapfianus have been successfully used to enhance drought and salt tolerance in model plants as well as important crop species. In addition to the ability to desiccate and rehydrate successfully, the survival of resurrection plants in regions experiencing short sporadic rainfall events may depend substantially on the ability to tightly down-regulate cell division and cell wall loosening activities with decreasing water availability and then grow rapidly after rainfall while water is plentiful. Hence, an analysis of gene transcripts present in the desiccated tissue of resurrection plants may reveal important growth-related genes. Recent findings support the proposition that, as well as being a versatile model for devising strategies for protecting plants from water-loss, resurrection plants may be a very useful tool for pinpointing genes to target for enhancing growth rate and biomass production.

Sporobolus stapfianus, a model desiccation-tolerant grass

Sporobolus stapfianus Gandoger, one of ~40 known ‘anabiotic’grass species (i.e. ‘able to regain vital activity from a state of latent life’), is the most versatile tool for research into desiccation tolerance in vegetative grass tissue. Current knowledge on this species is presented, including the features that suit it for investigations into the plant’s ability to survive dehydration of its leaf protoplasm. The main contributors to desiccation tolerance in S. stapfianus leaves appear to be: accumulation during dehydration of protectants of membranes and proteins; mechanisms limiting oxidative damage; a retention of protein synthetic activity in late stages of drying that is linked with changes in gene expression and in the proteomic array; and an ability to retain net synthesis of ATP during drying. S. stapfianus exemplifies an advanced stage of an evolutionary trend in desiccation tolerant plants towards increased importance of the dehydration phase (for induction of tolerance, for synthesis of protectants and for proteomic changes).

Real-time measurement of phloem turgor pressure in Hevea brasiliensis with a modified cell pressure probe

Background: Although the pressure flow theory is widely accepted for the transport of photoassimilates in phloem sieve elements, it still requires strong experimental validation. One reason for that is the lack of a precise method for measuring the real-time phloem turgor pressure from the sink tissues, especially in tree trunks. Results: Taking the merits of Hevea brasiliensis, a novel phloem turgor pressure probe based on the state of the art cell pressure probe was developed. Our field measurements showed that the phloem turgor pressure probe can sensitively measure the real-time variation of phloem turgor pressure in H. brasiliensis but the calculation of phloem turgor pressure with xylem tension, xylem sap osmotic potential and phloem sap osmotic potential will under-estimate it. The measured phloem turgor pressure gradient in H. brasiliensis is contrary to the Münch theory. The phloem turgor pressure of H. brasiliensis varied from 8-12 bar as a consequence of water withdrawal from transpiration. Tapping could result in a sharp decrease of phloem turgor pressure followed by a recovery from 8-45 min after the tapping. The recovery of phloem turgor pressure after tapping and its change with xylem sap flow suggest the importance of phloem water relationship in the phloem turgor pressure regulation. Conclusion: The phloem turgor pressure probe is a reliable technique for measuring the real-time variation of phloem turgor pressures in H. brasiliensis. The technique could probably be extended to the accurate measurement of phloem turgor pressure in other woody plants which is essential to test the Münch theory and to investigate the phloem water relationship and turgor pressure regulation. © 2014 An et al.

A review of computational fluid dynamics applications in pressure-driven membrane filtration

Ongoing advances in computational performance and numerics have led to computational fluid dynamics (CFD) becoming a ubiquitous modelling tool. However, CFD methods have only been adopted to simulate pressure-driven membrane filtration systems relatively recently. This paper reviews various approaches to describing the behaviour of these systems using CFD, beginning with the hydrodynamics of membrane channels, including discussion of laminar, turbulent, and transition flow regimes, with reference to the effects of osmotic pressure, concentration polarisation, and cake formation. The use of CFD in describing mass transfer through the membrane itself is then discussed, followed by some concluding comments on commercial membrane simulation packages and future research directions in membrane CFD. © 2013 Springer Science+Business Media Dordrecht.

Comparison between nanofiltration and forward osmosis in the treatment of dye solutions

Colour removal and the flux behaviour of nanofiltration (NF-DOW FILMTEC-NF245) and forward osmosis (FO-a flat sheet cellulose triacetate membrane with a woven embedded backing support) membranes were investigated in this study. The NF membrane was employed to perform dye removal experiments with aqueous solutions containing 15 g/L of NaCl and different concentrations of Acid Green 25, Remazol Brilliant Orange FR and Remazol Blue BR dyes. The increase in dye concentration resulted in a decline in water permeability and an increase in colour removal. When the concentrations of dye solutions varied from 250 to 1000 mg/L, at 0.8 bar of trans-membrane pressure, the NF system exhibited a steady permeate flux of more 30 L/m2h and a colour removal of more than 99%; salt rejection was more than 20.0%. Furthermore, the FO system possessed high dye rejection efficiency (almost 100%), with low permeate flux of around 2.0 L/m2h, when using dye solutions as feed streams and seawater as draw stream. The mode of operation (either FO or pressure retarded osmosis (PRO) did not change the flux significantly but PRO mode always produced higher fluxes than FO mode under the operating conditions used in this study. While both NF and FO can be used to reduce the volume of effluent containing dyes from textile industries, the energy spent in NF on applied pressure can be substituted by the osmotic pressure of draw solution in FO when concentrated draw solutions such as sea water or reverse osmosis concentrate are readily available.

Latex dilution reaction during the tapping flow course of Hevea brasiliensis and the effect of Ethrel stimulation

The latex dilution reaction during the tapping flow course has been well documented and associated with the facilitation of tapping latex flow. However, its underlying mechanism has not experimentally examined. The latex total solid content, osmotic potential and phloem turgor pressure change during the tapping flow course were simultaneously measured to investigate the cause of water movement during the tapping flow course. It was found that there are three different stages for the laticifer water equilibrium during the tapping flow course. The tapping-induced rapid turgor pressure drop is the cause of the first stage water influx into laticifers, while osmoregulation prevails during water exchange in the second and third stages of tapping flow. Meanwhile, aquaporin expressions were, for the first time, investigated during the tapping flow course. The rapid transcript up-regulation of HbPIP1, HbPIP2;1 and HbPIP2;3 contributes to the latex dilution reaction. However, their activity gating cannot be ruled out. Ethrel stimulation can significantly dilute the corresponding latex fractions during the tapping flow course due to its up-regulations of HbPIP1, HbPIP2;1 and HbPIP2;3. Nevertheless, the latex dilution reaction pattern for the Ethrel treated trees did not change, except for a lower degree of dilution compared with the un-treated trees. All these results suggest that both phloem turgor pressure and aquaporins are involved in the latex dilution reaction during the tapping flow course.

Migrating swans profit from favourable changes in wind conditions at low altitude

Because energy reserves limit flight range, wind assistance may be of crucial importance for migratory birds. We tracked eight Bewick's swans Cygnus columbianus bewickii, using 95-g satellite transmitters with altimeters and activity sensors, during their spring migration from Denmark to northern Russia in 1996. During the 82 occasions where a swan's location was recorded in flight, average flight altitude was 165 m a.s.1. with a maximum of 759 m a.s.1., despite winds often being more favourable at higher altitudes. We also counted Bewick's swans departing from the Gulf of Finland and subsequently passing an observatory in the next major stop-over area 800 km further north in the White Sea, northern Russia, during the springs of 1994, 1995 and 1996. A comparison of these counts with wind data provided evidence for Bewick's swans using favourable changes in wind conditions to embark on migration. Changes in the numbers of birds arriving in the White Sea correlated best with favourable changes in winds in the Gulf of Finland 1 day earlier. Again, migratory volume showed a correlation with winds at low altitudes only, despite wind conditions for the swans being more favourable at high altitudes. We conclude that the relatively large Bewick's swan tends to gear its migration to wind conditions at low altitude only. We argue that Bewick's swans do not climb to high altitudes because of mechanical and physiological limitations with respect to the generation of power for flight and to avoid rapid dehydration.

Flight costs and fuel composition of a bird migrating in a wind tunnel

We studied the energy and protein balance of a Thrush Nightingale Luscinia luscinia, a small long-distance migrant, during repeated 12-hr long flights in a wind tunnel and during subsequent two-day fueling periods. From the energy budgets we estimated the power requirements for migratory flight in this 26 g bird at 1.91 Watts. This is low compared to flight cost estimates in birds of similar mass and with similar wing shape. This suggests that power requirements for migratory flight are lower than the power requirements for nonmigratory flight. From excreta production during flight, and nitrogen and energy balance during subsequent fueling, the dry protein proportion of stores was estimated to be around 10%. A net catabolism of protein during migratory flight along with that of fat may reflect a physiologically inevitable process, a means of providing extra water to counteract dehydration, a production of uric acid for anti-oxidative purposes, and adaptive changes in the size of flight muscles and digestive organs in the exercising animal.

Predicting migratory flight altitudes by physiological migration models

Using the altitudinal profiles of wind, temperature, pressure, and humidity in three flight models, we tried to explain the altitudinal distributions of nocturnal migrants recorded by radar above a desert in southern Israel. In the simplest model, only the tailwind component was used as a predictor of the most preferred flight altitude (T model). The energy model (E model) predicted flight ranges according to mechanical power consumption in flapping flight depending on air density and wind conditions, assuming optimal adjustment of airspeed and compensation of crosswinds, and including the influence of mass loss during flight. The energy-water model (EW model) used the same assumptions and parameters as the E model but also included restrictions caused by dehydration. Because wind was by far the most important factor governing altitudinal distribution of nocturnal migrants, differences in predictions of the three models were small. In a first approach, the EW model performed slightly better than the E model, and both performed slightly better than the T model. Differences were most pronounced in spring, when migrants should fly high according to wind conditions, but when climbing and descending they must cross lower altitudes where conditions are better with respect to dehydration. A simplified energy model (Es model) that omits the effect of air density on flight costs explained the same amount of variance in flight altitude as the more complicated E and EW models. By omitting the effect of air density, the Es model predicted lower flight altitudes and thus compensated for factors that generally bias height distributions downward but are not considered in the models (i.e. climb and descent through lower air layers, cost of ascent, and decrease of oxygen partial pressure with altitude). Our results confirm that wind profiles, and thus energy rather than water limitations, govern the altitudinal distribution of nocturnal migrants, even under the extreme humidity and temperature conditions in the trade wind zone.

Fluid intake, hydration, work physiology of wildfire fighters working in the heat over consecutive days

PURPOSE: (i) To evaluate firefighters' pre- and post-shift hydration status across two shifts of wildfire suppression work in hot weather conditions. (ii) To document firefighters' fluid intake during and between two shifts of wildfire suppression work. (iii) To compare firefighters' heart rate, activity, rating of perceived exertion (RPE), and core temperature across the two consecutive shifts of wildfire suppression work. METHOD: Across two consecutive days, 12 salaried firefighters' hydration status was measured immediately pre- and post-shift. Hydration status was also measured 2h post-shift. RPE was also measured immediately post-shift on each day. Work activity, heart rate, and core temperature were logged continuously during each shift. Ten firefighters also manually recorded their food and fluid intake before, during, and after both fireground shifts. RESULTS: Firefighters were not euhydrated at all measurement points on Day one (292±1 mOsm l(-1)) and euhydrated across these same time points on Day two (289±0.5 mOsm l(-1)). Fluid consumption following firefighters' shift on Day one (1792±1134ml) trended (P = 0.08) higher than Day two (1108±1142ml). Daily total fluid intake was not different (P = 0.27), averaging 6443±1941ml across both days. Core temperature and the time spent ≥ 70%HRmax were both elevated on Day one (when firefighters were not euhydrated). Firefighters' work activity profile was not different between both days of work. CONCLUSION: There was no difference in firefighters' pre- to post-shift hydration within each shift, suggesting ad libitum drinking was at least sufficient to maintain pre-shift hydration status, even in hot conditions. Firefighters' relative hypohydration on Day one (despite a slightly lower ambient temperature) may have been associated with elevations in core temperature, more time in the higher heart rate zones, and 'post-shift' RPE.

Comparison of cell disruption methods for improving lipid extraction from thraustochytrid strains

Lipid extraction is an integral part of biodiesel production, as it facilitates the release of fatty acids from algal cells. To utilise thraustochytrids as a potential source for lipid production. We evaluated the extraction efficiency of various solvents and solvent combinations for lipid extraction from Schizochytrium sp. S31 and Thraustochytrium sp. AMCQS5-5. The maximum lipid extraction yield was 22% using a chloroform:methanol ratio of 2:1. We compared various cell disruption methods to improve lipid extraction yields, including grinding with liquid nitrogen, bead vortexing, osmotic shock, water bath, sonication and shake mill. The highest lipid extraction yields were obtained using osmotic shock and 48.7% from Schizochytrium sp. S31 and 29.1% from Thraustochytrium sp. AMCQS5-5. Saturated and monounsaturated fatty acid contents were more than 60% in Schizochytrium sp. S31 which suggests their suitability for biodiesel production.

Artificial water points facilitate the spread of an invasive vertebrate in arid Australia

Summary: The spread of invasive species after their initial introduction is often facilitated by human actions. In some cases, invaders only become established in habitats where dominant native species have been displaced as a result of human actions or where humans inadvertently provide essential resources such as food, water or shelter. We investigated if dams that provide water for livestock have facilitated the cane toad's (Rhinella marina) invasion of a hot semi-arid landscape by providing toads with a resource subsidy and hence refuge from extreme heat and aridity. To determine the relationship between the presence of surface water and habitat occupancy by toads, we surveyed natural and artificial water features for cane toads during the annual dry season. We used radiotracking and acoustic tags to determine whether movement patterns and shelter use of cane toads were focussed around dams. To determine whether dams provide toads with refuge from extreme heat and aridity, we deployed plaster models with internal thermometers to estimate ambient temperatures and toad desiccation rates in shelter sites. To determine whether dams alleviate the stress experienced by toads, we measured plasma corticosterone levels of toads that sheltered in and away from dams. Toads were present in sites with standing water and absent from waterless sites. Most radiotracked toads sheltered within 1 m of water. Toad movements were focussed around water. Toads tracked with passive acoustic telemetry over a 6-month dry season were highly resident at dams. Plaster models placed in toad shelter sites away from the water lost 27% more mass and experienced higher temperatures than models placed near the water's edge. Toads that sheltered in terrestrial shelters exhibited higher plasma corticosterone levels compared to toads that sheltered near dams. Dams provide toads with refuge habitats where they are less at risk from overheating and dehydration. Synthesis and applications. Artificial water points can facilitate biological invasions in arid regions by providing a resource subsidy for water-dependent invasive species. Our study suggests that there is scope to control populations of water-dependent invasive vertebrates in arid regions by restricting their access to artificial water points.

Adrenocortical stress responses influence an invasive vertebrate's fitness in an extreme environment

Continued range expansion into physiologically challenging environments requires invasive species to maintain adaptive phenotypic performance. The adrenocortical stress response, governed in part by glucocorticoid hormones, influences physiological and behavioural responses of vertebrates to environmental stressors. However, any adaptive role of this response in invasive populations that are expanding into extreme environments is currently unclear. We experimentally manipulated the adrenocortical stress response of invasive cane toads (Rhinella marina) to investigate its effect on phenotypic performance and fitness at the species' range front in the Tanami Desert, Australia. Here, toads are vulnerable to overheating and dehydration during the annual hot-dry season and display elevated plasma corticosterone levels indicative of severe environmental stress. By comparing unmanipulated control toads with toads whose adrenocortical stress response was manipulated to increase acute physiological stress responsiveness, we found that control toads had significantly reduced daily evaporative water loss and higher survival relative to the experimental animals. The adrenocortical stress response hence appears essential in facilitating complex phenotypic performance and setting fitness trajectories of individuals from invasive species during range expansion.

Transformação genética em cevada por bombardeamento de partículas

A presente Tese de Doutorado objetivou: (1) definir um método eficiente de transformação genética, por bombardeamento de partículas, para a obtenção de plantas transgênicas de cultivares brasileiras de cevada e (2) identificar gene(s) codificante(s) de quitinase(s) potencialmente capaz(es) de conferir resistência ao fungo patogênico de cevada Bipolaris sorokiniana. Culturas de calos obtidos a partir de escutelos imaturos das cultivares Brasileiras de cevada MN-599 e MN-698 (Cia. de Bebidas das Américas, AMBEV) foram bombardeadas com partículas de tungstênio e avaliadas quanto à expressão do gene repórter gusA através de ensaios histoquímicos de GUS e quanto ao efeito dos bombardeamentos na indução estruturas embriogênicas e regeneração de plantas. As condições de biobalística analisadas incluíram a região promotora regulando a expressão de gusA, tipo e pressão de gás hélio de dois aparelhos de bombardeamento, distância de migração das partículas, número de tiros e a realização de pré e pós-tratamento osmótico dos tecidos-alvo. No presente trabalho foram obtidos um número bastante alto de pontos azuis por calo, a indução de calos embriogênicos e embriões somáticos em uma freqüência de até 58,3% e a regeneração de 60 plantas, sendo 43 de calos bombardeados. As melhores condições observadas foram o promotor e primeiro íntron do gene Adh de milho (plasmídeo pNGI), o aparelho de bombardeamento “ Particle Inflow Gun” (PIG) utilizando-se a distância de migração de partículas de 14,8 cm, dois tiros disparados por placa e a realização de tratamento osmótico dos explantes com 0,2 M de manitol e 0,2 M de sorbitol 4-5 horas antes e 17-19 horas depois dos bombardeamentos. Das 43 plantas obtidas de calos bombardeadas, 3 apresentaram atividade de GUS na base das suas folhas. A utilização de primers sintéticos definidos a partir de genes de quitinases descritos na literatura em PCRs resultou na amplificação de dois fragmentos de aproximadamente 700 e 500 pb a partir de DNA total das cvs. MN-599 e MN-698 de cevada e um fragmento, com aproximadamente 500 pb, a partir do DNA total do isolado A4c de Trichoderma sp. Estes fragmentos foram purificados dos géis de agarose e diretamente seqüenciados de forma manual e automática. Os fragmentos de 700 e 500 pb amplificados do genoma da cultivar MN-599 foram identificados como genes de quitinases de cevada e o fragmento de 500 pb do isolado A4c de Trichoderma sp. não apresentou homologia com seqüências conhecidas de quitinases depositadas no EMBL/GenBank. A utilização de novos pares de primers, representando seqüências conservadas de quitinases do fungo Metarhizium anisopliae, resultou na amplificação de 3 fragmentos a partir do DNA total do isolado A4b de Trichoderma sp., que estão sendo purificados para realização de seqüenciamento.

Vitrificação de embriões Mus domesticus domesticus contidos em volumes diferentes de 9,0 m de etileno glicol.

Os experimentos tiveram como objetivo determinar a taxa de eclosão dos embriões vitrificados em volumes diferentes de 9,0 M de etileno glicol. Simultaneamente, testou-se dois procedimentos de estocagem dos fios de teflon, denominados caixa de aço inoxidável e globete/raque. No experimento I, os 881 embriões coletados foram distribuídos em 4 tratamentos: tratamento 1 (T1= controle): 307 embriões foram cultivados in vitro em meio PBSm, acrescido de 0,4% de BSA; tratamento 2 (T2): 292 embriões foram expostos à solução de glicerol 10% acrescida de 0,4% de BSA, envasados em palhetas de 0,25 mL e submetidos ao congelamento pelo método rápido em Biocool; tratamento 3 (T3): 138 embriões foram expostos durante 2 minutos à solução de desidratação (10% de EG + 6% BSA em PBSm) e então transferidos para a solução de vitrificação (50% de EG + 6% de BSA em PBSm), onde permaneceram por 30 segundos e foram colocados em volume de 1 μL no interior de um fio de teflon, medindo 0,4 mm de diâmetro, 2,0 cm de comprimento e 0,05 mm de espessura. Os fios foram acondicionados em uma caixa de aço inoxidável para serem armazenados em nitrogênio líquido; tratamento 4 (T4): 144 embriões foram expostos à solução de desidratação (10% de EG + 6% BSA em PBSm) e após 2 minutos, foram transferidos para a solução de vitrificação (50% de EG + 6% BSA em PBSm), onde permaneceram por 30 segundos, sendo após transferidos para um volume de 1 μL no interior do fio de teflon. Os fios de teflon foram estocados em globetes unidos às raques e mantidos em nitrogênio líquido. Após o aquecimento, os embriões foram cultivados em PBSm suplementado com 0,4% de BSA. As taxas de eclosão embrionária observadas foram: T1=76,29% (245/307); T2=41,05% (117/292); T3=37,98% (54/138) e T4=26,78% (37/144). No segundo experimento, 747 embriões foram distribuídos em 3 tratamentos: tratamento 1 (T1= controle): 80 embriões foram cultivados in vitro em meio KSOM acrescido de 0,4% de BSA; tratamento 2 (T2): 334 embriões expostos em solução de glicerol 10% acrescida de 0,4% de BSA, foram envasados em palhetas de 0,25 mL e submetidos ao congelamento pelo método rápido em Biocool; tratamento 3 (T3): 333 blastocistos foram expostos durante 2 minutos à solução de desidratação (10% de EG + 0,4% BSA em PBSm) e então transferidos para tubos eppendorf de 2,0 mL em contato com a solução de vitrificação (50% de EG + 0,4% BSA em PBSm). Após o cultivo in vitro, as taxas de eclosão embrionária observadas nos 3 tratamentos foram respectivamente: 88,75% (71/80), 40,44% (141/334) e 19,70% (66/333). Baseado nesses resultados conclui-se que embriões Mus domesticus domesticus submetidos à técnica de vitrificação após exposição à solução de 9,0 M de etileno glicol e envase em fios de teflon assegurou índices satisfatórios de sobrevivência embrionária. As taxas de sobrevivência dos embriões Mus domesticus domesticus foi independente do procedimento de estocagem em botijão de nitrogênio líquido. A vitrificação em solução de 9,0 M de etileno glicol com envase em tubos eppendorf não foi eficiente para promover altas taxas de sobrevivência embrionária, mas proporcionou segurança biológica aos embriões, durante o armazenamento.