Characterization of rhizospheric bacteria isolated from Deschampsia antarctica Desv.

Deschampsia antarctica Desv. is the only gramineae capable of colonizing the Antarctic due to the region’s extreme climate and soil environment. In the present research, bacteria colonizing the rhizospheric soil of D. antarctica were isolated and characterized. The soil studies showed that D. antarctica possesses a wide spectrum of psychrotolerant bacteria with extensive and varied antibiotic resistance, as well as heavy metal tolerance. The bacterial strains isolated from the rhizosphere of D. antarctica also produced a diverse pattern of enzymes. Based on the strain identification with partial characterization of the 16S rRNA gene, the majority of the isolates correspond to different Pseudomonas species, and species of the genus Flavobacterium sp. and Arthrobacter sp. The isolated strains collected from this research constitute a unique collection for future, more detailed taxonomic analysis and physiological characterization, contributing to the search for potential biotechnological uses. These findings and others have great potential for developing new biotechnological products from Antarctic microorganisms.     

Differential accumulation of dehydrin-like proteins by abiotic stresses in Deschampsia antarctica Desv.

Dehydrins are proteins that accumulate during environmental stresses leading to cell dehydration. Deschampsia antarctica is one of the two vascular plants that have colonized the Maritime Antarctic. This plant is usually exposed to cold, salt and desiccating winds in the field. We proposed that among the factors that allow D. antarctica to survive the harsh environmental conditions is the presence of dehydrins. We studied the accumulation of dehydrins by abscisic acid (ABA), dehydration, NaCl and low osmotic potential. Western blots using an anti-dehydrin antibody revealed a complex pattern of dehydrin-like proteins (DLPs) accumulation in the different treatments. DLPs with apparent molecular weight of 58, 57, 55, 53, 48, 42, 32, 30, 28 and 25 kDa were detected in the different treatments. DLPs accumulation was associated with a decrease in the relative water content (RWC) of the plants. These results suggest that DLPs accumulation could contribute to explain how D. antarctica can survive under adverse Antarctic conditions.     

New ultrastructural features of organelles in leaf cells of Deschampsia antarctica Desv

An electron microscope has been used to investigate the ultrastructure of leaf cells in Deschampsia antarctica Desv. (Poaceae). The leaf anatomy exhibits features typical of xerophytes. New ultrastructural features were found in mesophyll cells. Chloroplasts in mesophyll cells of D. antarctica leaves form small vesicles and pockets. The outer chloroplast membrane forms vesicles, and pockets are invaginations of both membranes. The invaginations contain small vesicles, mitochondria, or lipid droplets. The mitochondria or peroxisomes adhere very tightly to the chloroplasts.     

Regional genetic diversity patterns in Antarctic hairgrass (Deschampsia antarctica Desv.)

Aim  To determine patterns in diversity of a major Antarctic plant species, including relationships of Antarctic populations with those outside the Antarctic zone.
Location  Antarctic Peninsula, Maritime Antarctica, sub-Antarctic islands, Falkland Islands and South America.
Methods  Amplified fragment length polymorphisms (AFLPs) and chloroplast sequences were used to study patterns of genetic diversity in Antarctic hairgrass ( Deschampsia antarctica Desv.) and the genetic relationships between populations over its distribution range. Thirty-eight populations were sampled from a large part of the distribution of D. antarctica , and additionally, herbarium specimens were included for areas from which we could not obtain fresh samples.
Results  A gradient in AFLP diversity was observed going from the Falklands southwards into the Antarctic. This gradient in diversity was also observed within the Antarctic Peninsula: diversity was lower further south. Diversity in the chloroplast genome of D. antarctica was low. Only three chloroplast haplotypes were found, each with a strong regional distribution.
Main conclusions  The phylogenetic construction of AFLP marker frequencies in meta-populations of D. antarctica supports a stepping-stone model of colonization, whereby gene flow mainly occurs between neighbouring populations. It is concluded that long-distance gene flow is very limited in D. antarctica . A very low diversity was found in the sub-Antarctic islands in the Indian Ocean, indicating that these populations have experienced a recent evolutionary bottleneck.     

Deschampsia antarctica Desv. primary photochemistry performs differently in plants grown in the field and laboratory

Primary photochemistry of photosystem II (F _v/F _m) of the Antarctic hair grass Deschampsia antarctica growing in the field (Robert Island, Maritime Antarctic) and in the laboratory was studied. Laboratory plants were grown at a photosynthetic photon flux density (PPFD) of 180 μmol m⁻² s⁻¹ and an optimal temperature (13 ± 1.5°C) for net photosynthesis. Subsequently, two groups of plants were exposed to low temperature (4 ± 1.5°C day/night) under two levels of PPFD (180 and 800 μmol m⁻² s⁻¹) and a control group was kept at 13 ± 1.5°C and PPFD of 800 μmol m⁻² s⁻¹. Chlorophyll fluorescence was measured during several days in field plants and weekly in the laboratory plants. Statistically significant differences were found in F _v/F _m (=0.75–0.83), F ₀ and F _m values of field plants over the measurement period between days with contrasting irradiances and temperature levels, suggesting that plants in the field show high photosynthetic efficiency. Laboratory plants under controlled conditions and exposed to low temperature under two light conditions showed significantly lower F _v/F _m and F _m. Moreover, they presented significantly less chlorophyll and carotenoid content than field plants. The differences in the performance of the photosynthetic apparatus between field- and laboratory-grown plants indicate that measurements performed in ex situ plants should be interpreted with caution.     

Retrotransposon‐based genetic diversity of Deschampsia antarctica Desv. from King George Island (Maritime Antarctic)

Deschampsia antarctica Desv. can be found in diverse Antarctic habitats which may vary considerably in terms of environmental conditions and soil properties. As a result, the species is characterized by wide ecotypic variation in terms of both morphological and anatomical traits. The species is a unique example of an organism that can successfully colonize inhospitable regions due to its phenomenal ability to adapt to both the local mosaic of microhabitats and to general climatic fluctuations. For this reason, D. antarctica has been widely investigated in studies analyzing morphophysiological and biochemical responses to various abiotic stresses (frost, drought, salinity, increased UV radiation). However, there is little evidence to indicate whether the observed polymorphism is accompanied by the corresponding genetic variation. In the present study, retrotransposon‐based iPBS markers were used to trace the genetic variation of D. antarctica collected in nine sites of the Arctowski oasis on King George Island (Western Antarctic). The genotyping of 165 individuals from nine populations with seven iPBS primers revealed 125 amplification products, 15 of which (12%) were polymorphic, with an average of 5.6% polymorphic fragments per population. Only one of the polymorphic fragments, observed in population 6, was represented as a private band. The analyzed specimens were characterized by low genetic diversity (uH_e = 0.021, I = 0.030) and high population differentiation (F_ST = 0.4874). An analysis of Fu's F_S statistics and mismatch distribution in most populations (excluding population 2, 6 and 9) revealed demographic/spatial expansion, whereas significant traces of reduction in effective population size were found in three populations (1, 3 and 5). The iPBS markers revealed genetic polymorphism of D. antarctica, which could be attributed to the mobilization of random transposable elements, unique features of reproductive biology, and/or geographic location of the examined populations.     

The influence of some environmental factors on cytological and biometric parameters and chlorophyll content of Deschampsia antarctica Desv. in the maritime Antarctic

Under the environmental conditions of the Point Thomas Oasis (King George Island, the South Shetland Islands), we studied the influence of month-long artificial treatment with fresh water, salt water, and guano solution on the biometric characteristics, chlorophyll content, as well as the nuclear area of leaf parenchymal cells and nuclear DNA content, in a maritime Antarctic aboriginal plant Deschampsia antarctica. The modeled factors induced an increase in the generative shoot height and the length of the largest leaf, but did not influence the number of flowers. Treatment with guano caused an increase in the chlorophyll a and b contents, while fresh water treatment only led to some increase in chlorophyll a. Fluctuations of physiologically significant traits, such as the nuclear area and DNA content in the leaf parenchyma cells of D. antarctica, have been traced under the influence of the studied factors. Understanding of the hierarchy of influence of these factors as well as and sensitivity of plants of this species to external agents require further investigation.     

Leishmanicidal and antitumoral activities of endophytic fungi associated with the Antarctic angiosperms Deschampsia antarctica Desv. and Colobanthus quitensis (Kunth) Bartl.

A total of 564 isolates of endophytic fungi were recovered from the plants Deschampsia antarctica and Colobanthus quitensis collected from Antarctica. The isolates were screened against parasites Leishmania amazonensis and Trypanosoma cruzi and against the human tumour cell lines. Of the 313 fungal isolates obtained from D. antarctica and 251 from C. quitensis, 25 displayed biological activity. Nineteen extracts displayed leishmanicidal activity, and six inhibited the growth of at least one tumour cell line. These fungi belong to 19 taxa of the genera Alternaria, Antarctomyces, Cadophora, Davidiella, Helgardia, Herpotrichia, Microdochium, Oculimacula, Phaeosphaeria and one unidentified fungus. Extracts of 12 fungal isolates inhibited the proliferation of L. amazonesis at a low IC₅₀ of between 0.2 and 12.5 μg ml⁻¹. The fungus Phaeosphaeria herpotrichoides displayed only leishmanicidal activity with an IC₅₀ of 0.2 μg ml⁻¹, which is equivalent to the inhibitory value of amphotericin B. The extract of Microdochium phragmitis displayed specific cytotoxic activity against the UACC-62 cell line with an IC₅₀ value of 12.5 μg ml⁻¹. Our results indicate that the unique angiosperms living in Antarctica shelter an interesting bioactive fungal community that is able to produce antiprotozoal and antitumoral molecules. These molecules may be used to develop new leishmanicidal and anticancer drugs.     

Characterization of a novel antarctic plant growth-promoting bacterial strain and its interaction with antarctic hair grass (Deschampsia antarctica Desv)

Deschampsia antarctica is the only hair grass that has been able to successfully colonize the Antarctic continent. However, there is little research on the role of microorganisms associated with the rhizosphere that may participate in its growth and development. The objective of this research was to characterize a psychrotolerant bacterial strain isolated from the rhizosphere of D. antarctica. Biochemical and molecular studies were performed to characterize this bacterium. It was determined that this strain secretes a neutral polysaccharide that presents different compositions at different temperatures (4 and 20 °C). Based on biochemical and phylogenetic analyses, the Antarctic rhizobacterium could be a new species of Pseudomonas. To determine their ability to solubilize different sources of inorganic phosphate, qualitative and quantitative analyses were conducted to determine P released at 4 °C. The Antarctic strain of Pseudomonas sp. was able to solubilize all sources of phosphates, and 34.2 mg P/L was released from rock phosphate. Growth physiological parameters were evaluated for seedlings of D. antarctica inoculated with the rhizobacteria. It was found that the bacterial inoculation promoted plant root development. SEM analysis of the roots showed that the bacterium is mainly located in the root hairs of D. antarctica.     

Effects of salinity and temperature on Deschampsia antarctica

Deschampsia antarctica is one of two species of vascular plants native to Antarctica. Populations of D. antarctica have become established on recently exposed glacial forelands on the Antarctic Peninsula and these plants may rely upon nutrient inputs from hauled out mammals, seabirds and sea spray. However, not much is known about the ability of these plants to tolerate salinity stress. We examined the effects of salinity and temperature on growth, reproduction, chlorophyll fluorescence and water relations of D. antarctica. In addition, we analysed concentrations of free proline in leaves and roots as previous studies have found large increases in the concentration of this amino acid in response to environmental stress. The growth chamber experiment was a 3 × 3 (temperature × salinity) complete factorial. Plants were grown at three temperature regimes: 7°/7°C, 12°/7°C, and 20°/7°C day/night and three salinity levels: <0.02 decismen per metre (dS m⁻¹; “low salinity”), 2.5 dS m⁻¹ (“medium salinity”), and 5.0 dS m⁻¹ (“high salinity”) for 66 days. Warmer temperatures improved leaf and tiller production as well as leaf and root length, which is consistent with previous findings on this species. Salinity reduced final root length by 6 and 13% in the medium and high-salinity treatments, respectively. Plants growing in medium and high-salinity treatments had xylem pressures that were more negative and higher free-proline concentrations, suggesting that proline may act as an osmoregulant in D. antarctica.     

The effects of growth regulators and a scanning electron microscope study of somatic embryogenesis in Antartic hair grass (Deschampsia antarctica Desv.)

Deschampsia antartctica Desv. is a type of grass that is physiologically and biochemically adapted to the extreme environmental conditions of the Antarctic continent, which is of interest to many investigators. To explore the potential use of somatic embryogenesis as a biotechnological tool for the mass micropropagation of this grass, the effects of three dosages of 2,4-dichlorophenoxyacetic acid, dicamba, and picloram were evaluated. The developmental and morphological stages of somatic embryo formation were evaluated using scanning electron microscopy (SEM). Plant regeneration was evaluated under the effects of different dosages of 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA), alone and combined. The results indicated that a Murashige and Skoog basal medium supplemented with 3 mg/l of dicamba was the best for inducing somatic embryogenesis, while the combination of 1 mg/l BAP and 0.1 mg/l of NAA was the most efficient for the regeneration and development of the plants. This work demonstrates, for the first time with the use of SEM, that it is possible to apply somatic embryogenesis for the regeneration of superficial and morphological structures of somatic embryos in the species D. antarctica.     

Deschampsia antarctica: genetic and molecular-biological aspects of spreading in Antarctica

The genetical and molecular biological attributes of Deschampsia antarctica are considered in relation to its remarkable success in the colonization of the maritime Antarctic. However, none of the data from several published research studies provide convincing evidence that this species possess any obvious unique or specialised adaptations which might account for this success. While its occurrence appears to have spanned much of the Holocene, there is no evidence that there have ever been any other native vascular plant species in the maritime Antarctic. It is still unclear why no other taxa have succeeded in colonising this region. Further intensive studies of genetics and molecular biology of this species, and of other potential immigrants, may provide a better understanding of their enigmatic success in the maritime Antarctic.