The roots of the halophyte Salicornia brachiata are a source of new halotolerant diazotrophic bacteria with plant growth-promoting potential

Soil salinity is the major cause limiting plant productivity worldwide. Nitrogen-fixing bacteria were enriched and characterised from roots of Salicornia brachiata, an extreme halophyte which has substantial economic value as a bioresource of diverse and valuable products. Nitrogen-free semisolid NFb medium with malate as carbon source and up to 4% NaCl were used for enrichment and isolation of diazotrophic bacteria. The isolates were tested for plant growth-promoting traits and 16S rRNA, nifH and acdS genes were analysed. For selected strains, plant growth-promoting activities were tested in axenically grown Salicornia seedlings at different NaCl concentrations (0–0.5M). New halotolerant diazotrophic bacteria were isolated from roots of S. brachiata. The isolates were identified as Brachybacterium saurashtrense sp. nov., Zhihengliuella sp., Brevibacterium casei, Haererehalobacter sp., Halomonas sp., Vibrio sp., Cronobacter sakazakii, Pseudomonas spp., Rhizobium radiobacter, and Mesorhizobium sp. Nitrogen fixation as well as plant growth-promoting traits such as indole acetic acid (IAA) production, phosphate solubilisation, and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity were demonstrated. For Brachybacterium saurashtrense and Pseudomonas sp., significant plant growth-promoting activities were observed in Salicornia in salt stress conditions. Salicornia brachiata is a useful source of new halotolerant diazotrophic bacteria with plant growth-promoting potential.     

Variation in the natural abundance of 15N in the halophyte,Salicornia virginica,associated with groundwater subsidies of nitrogen in a southern California salt-marsh

To provide insight into the importance of the salt-marsh ecotone as a sink for inorganic nitrogen in perched groundwater, measurements were made of the natural abundance of ¹⁵N in dissolved NO₃-N and NH₄-N and in the salt-marsh halophyte, Salicornia virginica, along an environmental gradient from agricultural land into a salt-marsh. The increase in the natural abundance of ¹⁵N (expressed by convention as ¹⁵N) of NO₃-N, accompanied by the decrease in NO₃-N (and total dissolved inorganic N, DIN) concentration along the gradient, suggested that the salt-marsh ecotone is a site of transformation, most likely through denitrification, of inorganic nitrogen in groundwater. ¹⁵N enrichment in S. virginica (and the parasitic herb, Cuscuta salina), along the tidal marsh boundary, relative to high and middle marsh locations, indicated the retention of groundwater nitrogen as vegetative biomass. The correlation between ¹⁵N _Salicornia and ¹⁵N_NH4 suggested a preference for NH₄-N over NO₃-N during uptake by this plant. Groundwater inputs enhanced the standing crop, above-ground productivity, and nitrogen content of S. virginica but the ralative effects of pore water salinity and DIN concentration on these parameters were not determined. ¹⁵N enrichment of marsh plants by groundwater DIN inputs could prove useful in tracing the fate of these inputs in the marsh food web.     

Requirement of sodium chloride for the action of gibberellic acid in stimulating hypocotyl elongation of a halophyte, Salicornia herbacea L.

NaCl stimulated hypocotyl elongation of the halophyte Salicorniaherbacea L. grown either in light or dark. Its optimal concentrationwas around 0.1–0.2 M and its promoting effect was muchmore prominent in the dark. Gibberellic acid at 10^–5 Mstimulated hypocotyl elongation in light but not in the dark.Indole-3-acetic acid and kinetin were ineffective in promotinghypocotyl elongation. In light, gibberellic acid and NaCl synergisticallyenhanced hypocotyl elongation when both were given simultaneously.The action of NaCl could be replaced by KCl, but not by mannitol.Osmotic pressure of the epidermis of the Salicornia hypocotylincreased in response to gibberellic acid and/or NaCl treatment.Na⁺ content in the hypocotyl increased with NaCl application.Gibberellic acid and NaCl when given alone increased the extensibilityof the hypocotyl cell wall. Synergistic interaction in increasingthe extensibility was observed between gibberellic acid andNaCl. Stress-relaxation analysis of mechanical properties ofthe hypocotyl wall revealed that gibberellic acid and NaCl actedsynergistically in decreasing minimum relaxation time. Basedon these results, a possible mechanism by which gibberellicacid and NaCl regulate hypocotyl elongation of Salicornia herbaceaL., a typical halophilic plant, is discussed. ¹ Present address: Laboratory of Biology, Tezukayama College,Gakuen Minami, Nara 631, Japan. (Received June 13, 1978; )     

Cloning and transcript analysis of type 2 metallothionein gene (SbMT-2) from extreme halophyte Salicornia brachiata and its heterologous expression in E. coli

Aeromonas hydrophila is both a human and animal pathogen, and the cytotoxic enterotoxin (Act) is a crucial virulence factor of this bacterium because of its associated hemolytic, cytotoxic, and enterotoxic activities. Previously, to define the role of some regulatory genes in modulating Act production, we showed that deletion of a glucose-inhibited division gene (gidA) encoding tRNA methylase reduced Act levels, while overproduction of DNA adenine methyltransferase (Dam) led to a concomitant increase in Act-associated biological activities of a diarrheal isolate SSU of A. hydrophila. Importantly, there are multiple GATC binding sites for Dam within an upstream sequence of the gidA gene and one such target site in the act gene upstream region. We showed the dam gene to be essential for the viability of A. hydrophila SSU, and, therefore, to better understand the interaction of the encoding genes, Dam and GidA, in act gene regulation, we constructed a gidA in-frame deletion mutant of Escherichia coli GM28 (dam⁺) and GM33 (?dam) strains. We then tested the expressional activity of the act and gidA genes by using a promoterless pGlow-TOPO vector containing a reporter green fluorescent protein (GFP). Our data indicated that in GidA⁺ strains of E. coli, constitutive methylation of the GATC site(s) by Dam negatively regulated act and gidA gene expression as measured by GFP production. However, in the ?gidA strains, irrespective of the presence or absence of constitutively active Dam, we did not observe any alteration in the expression of the act gene signifying the role of GidA in positively regulating Act production. To determine the exact mechanism of how Dam and GidA influence Act, a real-time quantitative PCR (RT-qPCR) assay was performed. The analysis indicated an increase in gidA and act gene expression in the A. hydrophila Dam-overproducing strain, and these data matched with Act production in the E. coli GM28 strain. Thus, the extent of DNA methylation caused by constitutive versus overproduction of Dam, as well as possible conformation of DNA influence the expression of act and gidA genes in A. hydrophila SSU. Our results indicate that the act gene is under the control of both Dam and GidA modification methylases, and Dam regulates Act production via GidA.     

Optimization of processing conditions for the fractionation of triticale straw using pressurized low polarity water

Pressurized low polarity water (PLPW) fractionation of triticale straw was optimized to maximize hemicellulose and lignin yield, and to produce a cellulose rich fraction for biofuels production. The optimum PLPW conditions for hemicellulose yield was determined to be 165 °C, with a flow rate of 115 mL/min, and a solvent-to-solid ratio of 60 mL/g. Hemicellulose and lignin yield generally increased with increasing temperature and solvent-to-solid ratio. There was a small decrease in hemicellulose yield with an increase in flow rate. Minimum lignin content of the triticale straw residue after extraction was predicted to occur at a processing condition of 206 °C, 160 mL/min, and 67 mL/g. PLPW was successful in removing 73-78% of the hemicellulose, leaving a cellulose rich fraction (65% glucose concentration). Lignin was equally distributed between the solid residues and the extracts and most of the hemicellulose was extracted in oligomer form. Remaining solid residues after fractionation were highly digestible by cellulase enzymes.     

Photosynthetic limitations of a halophyte sea aster (Aster tripolium L) under water stress and NaCl stress

To understand the mechanisms of salt tolerance in a halophyte, sea aster ( Aster tripolium L.), we studied the changes of water relation and the factors of photosynthetic limitation under water stress and 300 mM NaCl stress. The contents of Na(+) and Cl(-) were highest in NaCl-stressed leaves. Leaf osmotic potentials ( Psi(s)) were decreased by both stress treatments, whereas leaf turgor pressure ( Psi(t)) was maintained under NaCl stress. Decrease in Psi(s) without any loss of Psi(t) accounted for osmotic adjustment using Na(+) and Cl(-) accumulated under NaCl stress. Stress treatments affected photosynthesis, and stomatal limitation was higher under water stress than under NaCl stress. Additionally, maximum CO(2) fixation rate and O(2) evolution rate decreased only under water stress, indicating irreversible damage to photosynthetic systems, mainly by dehydration. Water stress severely affected the water relation and photosynthetic capacity. On the other hand, turgid leaves under NaCl stress have dehydration tolerance due to maintenance of Psi(t) and photosynthetic activity. These results show that sea aster might not suffer from tissue dehydration in highly salinized environments. We conclude that the adaptation of sea aster to salinity may be accomplished by osmotic adjustment using accumulated Na(+) and Cl(-), and that this plant has typical halophyte characteristics, but not drought tolerance.     

Demography of the striped mouse (Rhabdomys pumilio) in the succulent karoo

The striped mouse (Rhabdomys pumilio) is widely distributed in southern Africa, inhabiting a wide range of habitats. We describe the demography of the striped mouse in the arid succulent karoo of South Africa, and compare our findings with those of published results for the same species from the moist grasslands of South Africa. In both habitats, breeding starts in spring, but the breeding season in the succulent karoo is only half as long as in the grasslands, which can be explained by different patterns and levels of rainfall; the succulent karoo receives mainly winter rain and rainfall is much less (about 160 mm year⁻¹) than in the grasslands (>1000 mm year⁻¹) which experience summer rain. Population density increased from 37 (start of breeding season) to 171 (end of breeding season) mice per hectare. A high yearly survival rate of 27% during our study from summer to the next breeding season resulted in a population density that was 10 times higher in the succulent karoo than in grasslands. The comparatively high population density may result in habitat saturation and thus forced philopatry, promoting group living in the succulent karoo, which contrasts with the solitary life-style exhibited by populations in moist grasslands.

Zusammenfassung

Demographie der Striemengrasmaus (Rhabdomys pumilio) in der SukkulentenkarooDie Striemengrasmaus (Rhabdomys pumilio) ist im südlichen Afrika weit verbreitet und kommt in unterschiedlichen Habitaten vor. In dieser Studie beschreiben wir die Demographie der Striemengrasmaus in der ariden Sukkulentenkaroo Südafrikas, und vergleichen unsere Ergebnisse mit den Ergebnissen früherer Studien, welche dieselbe Art in den feuchten Grasländern Südafrikas untersucht haben. Die Fortpflanzungssaison beginnt in beiden Habitaten im Frühling, aber sie ist in der Sukkulentenkaroo nur halb so lange wie in den Grasländern. Der Grund dafür wird im unterschiedlichen Niederschlagsmuster vermutet: In der Sukkulentenkaroo regnet es wenig im Winter (ca. 160 mm), während es in den Grasländern viel im Sommer regnet (>1000 mm). Die Überlebensrate in der Sukkulentenkaroo war vom Sommer zur nächsten Fortpflanzungssaison im Frühjahr mit 27% außerordentlich hoch, während die Populationsdichte zehnmal so groß war wie in den Grasländern. Am Anfang der Fortpflanzungssaison betrug die Populationsdichte 37 Mäuse ha⁻¹, am Ende der Fortpflanzungssaison 171 Mäuse ha⁻¹. Die außergewöhnlich hohe Populationsdichte führt womöglich zu Habitatsättigung und einem Fehlen freier Territorien, was mit erklären könnte, warum die Striemengrasmaus in der Sukkulentenkaroo gruppenlebend ist, in den Grasländern hingegen solitär.     

Morphological and physiological responses of the halophyte, Odyssea paucinervis (Staph) (Poaceae), to salinity

In this study, salt tolerance was investigated in Odyssea paucinervis Staph, an ecologically important C₄ grass that is widely distributed in saline and arid areas of southern Africa. Plants were subjected to 0.2%, 10%, 20%, 40%, 60% and 80% sea water dilutions (or 0.076, 3.8, 7.6, 15.2, 22.8, and 30.4 parts per thousand) for 11 weeks. Increase in salinity from 0.2% to 20% sea water had no effect on total dry biomass accumulation, while further increase in salinity to 80% sea water significantly decreased biomass by over 50%. Morphological changes induced by salinity included reductions in the number of culms, leaves and internodes as well as decreases in internode length and leaf length:leaf width ratios. Carbon dioxide exchange, leaf conductance and transpiration decreased at salinities of 40% and higher, while quantum yield of photosystem II (PSII), electron transport rate (ETR) through PSII and intrinsic photosynthetic efficiency generally decreased at salinities of 60% and higher compared to 0.2% sea water. Concentrations of Na⁺ and Cl⁻ increased significantly with salinity increase in both roots and shoots. Na⁺/K⁺ ratios in the roots and shoots ranged from 0.66 to 3.28 and increased with increase in substrate salinity. The maximal rate of secretion at 80% sea water was 415 nmol cm⁻² d⁻¹ for Na⁺ and 763 nmol m⁻² d⁻¹ for Cl⁻ with high selectivity for these two ions. Predawn and midday ψ decreased with increase in salinity and were more negative than those of the treatment solutions. The concentration of proline increased with increase in salinity in both roots and shoots. The data clearly indicated that O. paucinervis is a highly salt-tolerant species that is morphologically and physiologically adapted to a saline environment.     

Comparison of agarophytes (Gelidium, Gracilaria, and Gracilariopsis) as potential resources for bioethanol production

This study explores the possibility of producing ethanol using the acid hydrolysate of three abundant agar-containing red seaweeds (agarophytes): Gelidium amansii, Gracilaria tenuistipitata, and Gracilariopsis chorda. The main component in the seaweed samples was agar, which ranged from 20 to 51 % (g g^?1 dry weight). After optimizing acid hydrolysis, 100 g of seaweed was hydrolyzed at 130 °C for 15 min with 0.2 M H₂SO₄. Then, 120 mL of a 1:2 mixture of the hydrolysate broth and basal medium was fermented in a 200-mL bottle at 30 °C for 96 h. Of the three seaweeds, G. amansii had the best ethanol yield, producing 0.23 g g^?1 of galactose or 45 % of the theoretical yield. This yield increased to 60 % after detoxification of the hydrolysate with activated carbon.     

Steam pretreatment of H(2)SO(4)-impregnated Salix for the production of bioethanol

In the bioconversion of lignocellulosic materials to ethanol, pretreatment of the material prior to enzymatic hydrolysis is essential to obtain high overall yields of sugar and ethanol. In this study, steam pretreatment of fast-growing Salix impregnated with sulfuric acid has been investigated by varying the temperature (180-210 degrees C), the residence time (4, 8 or 12 min), and the acid concentration (0.25% or 0.5% (w/w) H(2)SO(4)). High sugar recoveries were obtained after pretreatment, and the highest yields of glucose and xylose after the subsequent enzymatic hydrolysis step were 92% and 86% of the theoretical, respectively, based on the glucan and xylan contents of the raw material. The most favorable pretreatment conditions regarding the overall sugar yield were 200 degrees C for either 4 or 8 min using 0.5% sulfuric acid, both resulting in a total of 55.6g glucose and xylose per 100g dry raw material. Simultaneous saccharification and fermentation experiments were performed on the pretreated slurries at an initial water-insoluble content of 5%, using ordinary baker's yeast. An overall theoretical ethanol yield of 79%, based on the glucan and mannan content in the raw material, was obtained.     

Cell growth and water relations of the halophyte,Atriplex nummularia L., in response to NaCl

Summary Growth reduction or cessation is an initial response of Atriplex nummularia L. cells to NaCl. However, A. nummularia L. cells that are adapted to 342 and 428 mM NaCl are capable of sustained growth in the presence of salt. Cells that are adapted to NaCl exhibit a reduced rate of division compared to unadapted cells. Unlike salt adapted cells of the glycophyte Nicotiana tabacum L., A. nummularia L. cells do not exhibit reduced rate of cell expansion after adaptation. However, the cell expansion rate of unadapted A. nummularia L. cells is considerably slower than that of unadapted glycophyte cells and this normally low rate of cell expansion may contribute to the enhanced capacity of the halophyte to tolerate salt. Turgor of NaCl adapted cells was equivalent to unadapted cells indicating that the cells of the halophyte do not respond to salt by osmotic over adjustment as reported for the glycophyte tobacco (Binzel et al. 1985, Plant Physiol. 79:118–125).     

Sediment phosphate fractionation and interstitial water phosphate concentration in two coastal lagoons (Albuferas de Adra,SE Spain)

The concentrations of o-phosphate and dissolved inorganic carbon were monitored in the interstitial water of the sediment in two highly eutrophic coastal lagoons, Laguna Honda and Laguna Nueva (SE, Spain) from July 2000 to August 2001. Additionally, the organic matter concentration and the P-fractions in the sediment were analysed. Despite their proximity, these two lagoons showed many significant differences (P<0.05). The P-concentration in the interstitial water of the top sediment was higher in the Laguna Honda (up to 1 mg l^–1), especially during summer, when both sites registered their highest values. By contrast, the concentration of organic matter and organic P-fractions were higher in the sediment of the Laguna Nueva. The concentration of the organic matter in the vertical profile of the sediment decreased with depth in the Laguna Nueva (from 17% at 0 cm to 7% at 15 cm, annual mean value) while it increased in the sediment of Laguna Honda (from 9% at 0 cm to 12% at 15 cm, annual mean value). The P-concentration in the interstitial water and the organic matter in the top sediment of Laguna Honda followed a seasonal pattern and, were both correlated with the temperature of the bottom water (r=0.706, P<0.05 and r= -0.929, P<0.01, respectively). The inorg-P fractions comprised 63% of the Tot-phosphate in the sediment of Laguna Honda, whereas org-P fractions (68%) dominated in the sediment of Laguna Nueva. The concentrations of Tot-P in the sestonic material collected at three different depths of the water column averaged 2.2 mg g^–1d.w. in Laguna Honda and 2.5 mg g^–1d.w. in Laguna Nueva. The average atomic C:P ratio of the sestonic material was significantly higher in Laguna Honda than in Laguna Nueva, but the top sediment in Laguna Nueva had a C:P ratio significantly higher than that of the sestonic material (average: 416 and 162, respectively).The different biodegradability of the organic matter in the top sediment probably explained the differences found in sediment composition between these two lagoons. We suggest that the P-reclying from the sediment to the interstitial water was faster in Laguna Honda due to the higher biodegradability of the organic matter. By contrast, the lower biodegradability of the organic matter in the top sediment of Laguna Nueva was probably due to a higher contribution of vascular plants which could explain the organic matter accumulation detected in this lagoon.     

Winter growth phenology and leaf orientation in Pachypodium namaquanum (Apocynaceae) in the succulent karoo of the Richtersveld,South Africa

Pachypodium namaquanum (Nyley ex Harb.) Welw., an unusual arborescent stem succulent from the succulent karoo of the arid Richtersveld in north-western South Africa and adjacent Namibia, is characterized by a striking curvature of the terminal 20–60 cm of the trunk toward the north. This orientation displays the single terminal whorl of drought-deciduous leaves with their flat surface angled at a mean inclination of 55° from horizontal. Inclination of 50–60° was found in 65% of individuals sampled, and 85% were inclined between 45 and 65°. Northward azimuth was also quite regular, but varied slightly between populations. The fixed leaf orientation in P. namaquanum maximizes radiation absorption during the winter months when leaves are present. Leaves normally form in early fall (April) and abscise early in spring (October). Growing season conditions in the Richtersveld are relatively mild, with mean maximum temperature dropping only to 21.6°C in July, the coldest month of the year. Frosts are rare. By the fixed orientation of its leaf whorl, P. namaquanum is able to maintain nearly twice the midwinter radiation absorptance that it would have with horizontal orientation. Over an annual cycle the angled leaves receive more radiation than would horizontal leaves for each of the 6 months in which they are present on the plant. This increased winter irradiance is hypothesized to singificantly increase net primary production by concentrating growth activities in winter months and allowing the species to remain dormant during the hyperarid conditions of the hot summer months. Midwinter flowering from apical buds in P. namaquanum may also be aided by its stem orientation. The evolution of this characteristic pattern of winter growth phenology and nodding stem orientation may have come about because of low but relatively regular autumn precipitation and moderate winter temperatures. Slow and regular growth of P. namaquanum leads to long lifespans which may reach 300 years or more.     

Camera-trapping density estimates suggest critically low population sizes for the Wet Tropics subspecies of the spotted-tailed quoll (Dasyurus maculatus gracilis)

Accurate estimates of distribution and population density are critical for the management of threatened species. This is particularly pertinent for mammalian predators, whose generally low population density, elusive nature, and large home range requirements make it difficult to detect declines. We aimed to refine population estimates of the northern spotted-tailed quoll (Dasyurus maculatus gracilis) in the Wet Tropics bioregion, to estimate the total number of adults, the likely size of subpopulations across the known distribution of the subspecies, and its associated conservation status. We performed targeted upland camera-trapping surveys from June 2017 to May 2019. To calculate population densities, we used a combination of the number of individuals identified from each survey and the mean maximum distance moved from three life history stages. We then extrapolated these estimates to modelled suitable habitat areas, refined by the camera-trapping surveys. Population sizes for the six defined subpopulations were estimated, and ranged from approximately 5 to 105 individuals. The total population was estimated to be 221 individuals. This total population estimate, and the estimates for each of the subpopulations, are lower than previous published estimates and are cause for concern. Given the low population estimates presented here and unresolved threats driving declines in some subpopulations, we suggest elevation of this subspecies to Critically Endangered under the EPBC Act.     

Development of a low input system for growing wheat (Triticum vulgare) in a permanent understorey of white clover (Trifolium repens)

In three field experiments carried out during 1989-91, a permanent sward of pure white clover (Trifolium repens) was established to provide a source of N for winter or spring wheat crops (Triticum vulgare) directly drilled into the legume. Spring-sown wheat failed to compete with the clover, but wheat sown in the autumn established successfully. N fertiliser was applied to all three experiments at rates of 0, 50 and 100 kg N ha“¹ measurements of grain and whole-crop silage yields were made. Yields were low for all treatments, probably because of the dry conditions prevailing and the low soil N status of the site used. Yield responses to fertiliser were significant, despite the contribution to plant nutrition that the clover was intended to make. A key feature of the work was that the clover survived successive cereal crops and could be grazed and used as an understorey for later crops. Further, response to fertiliser N diminished with a successive crop implying a build-up of available soil N, which measurements confirmed had occurred. Use of the system obviated the need to use pesticides, although reasons for the lack of pest damage were not clear.