Desiccation-sensitive Species Research Articles

Dehydration and rehydration differently affect photosynthesis and volatile monoterpenes in bryophytes with contrasting ecological traits.

Bryophytes desiccate rapidly when relative humidity decreases. The capacity to withstand dehydration depends on several ecological and physiological factors. Volatile organic compounds (VOCs) may have a role in enhancing tolerance to desiccating bryophytes. However, the functions of VOCs in bryophytes have received little attention so far. We aimed to investigate the impact of a dehydration-rehydration treatment on primary carbon metabolism and volatile terpenes (VTs) in three bryophytes with contrasting ecological traits: Vessicularia dubyana, Porella platyphylla and Pleurochaete squarrosa. First, we confirmed the desiccation sensitivity gradient of the species. Under fully hydrated conditions, the photosynthetic rate (A) was inversely associated with stress tolerance, with a lower rate in more tolerant species. The partial recovery of A in P. platyphylla and P. squarrosa after rehydration confirmed the desiccation tolerance of these two species. On the other hand, A did not recover after rehydration in V. dubyana. Regarding VT, each species exhibited a distinct VT profile under optimum hydration, with the highest VT pool found in the more desiccation-sensitive species (V. dubyana). However, the observed species-specific VT pattern could be associated with the ecological habitat of each species. P. squarrosa, a moss of dry habitats, may synthesize mainly non-volatile secondary metabolites as stress-defensive compounds. On the other hand, V. dubyana, commonly found submerged, may need to invest photosynthetically assimilated carbon to synthesize a higher amount of VTs to cope with transient water stress occurrence. Further research on the functions of VTs in bryophytes is needed to deepen our understanding of their ecological significance.

The Co-Occurrence of Physiological and Epicotyl Physiological Dormancy in Three Desiccation-Sensitive Castanopsis (Fagaceae) Acorns from China with Specific Reference to the Embryonic Axis Position

Ecological significance of dormancy in desiccation-sensitive seeds is poorly understood. Quercus exhibits mutually exclusive occurrence of physiological (PD) and epicotyl dormancy (ePD), with no reported co-occurrence or dormancy class in other genera. We aimed to understand the dormancy in three Castanopsis species and document desiccation sensitivity and germination patterns concerning the embryonic axis position. We hypothesized that Castanopsis acorns are recalcitrant and potentially dormant. Fresh and cold-stratified acorns of Castanopsis chinensis, Castanopsis purpurella, and Castanopsis sclerophylla were subjected to desiccation and germination. Seedling emergence and internal morphology was monitored following cold (CS) and warm (WS) stratification. Fresh acorns had radicles emerge only after CS but require WS for shoot emergence. Drying to 20% moisture content led to complete death. In C. purpurella and C. sclerophylla, the embryonic axis was near the scar, and germination occurred by cracking the pericarp near the scar, which contrasts with C. chinensis. Moderate drying relieved dormancy due to the mechanical resistance of the pericarp. All three acorns were desiccation-sensitive and dormant. This is the first explicit report on PD and ePD co-occurrence in desiccation-sensitive seeds, but literature surveys allow for inference of such coexistence. CS alleviated PD and WS relieved ePD. Winter temperatures break PD, and acorns germinate during spring, but shoot emergence is delayed until summer. Our results are instructive for research on the dormancy of desiccation-sensitive species and the reproduction of Fagaceae species in subtropical forests.

Back to the roots, desiccation and radiation resistances are ancestral characters in bdelloid rotifers

BackgroundBdelloid rotifers are micro-invertebrates distributed worldwide, from temperate latitudes to the most extreme areas of the planet like Antarctica or the Atacama Desert. They have colonized any habitat where liquid water is temporarily available, including terrestrial environments such as soils, mosses, and lichens, tolerating desiccation and other types of stress such as high doses of ionizing radiation (IR). It was hypothesized that bdelloid desiccation and radiation resistance may be attributed to their potential ability to repair DNA double-strand breaks (DSBs). Here, these properties are investigated and compared among nine bdelloid species collected from both mild and harsh habitats, addressing the correlation between the ability of bdelloid rotifers to survive desiccation and their capacity to repair massive DNA breakage in a phylogenetically explicit context. Our research includes both specimens isolated from habitats that experience frequent desiccation (at least 1 time per generation), and individuals sampled from habitats that rarely or never experienced desiccation.ResultsOur analysis reveals that DNA repair prevails in somatic cells of both desiccation-tolerant and desiccation-sensitive bdelloid species after exposure to X-ray radiation. Species belonging to both categories are able to withstand high doses of ionizing radiation, up to 1000 Gy, without experiencing any negative effects on their survival. However, the fertility of two desiccation-sensitive species, Rotaria macrura and Rotaria rotatoria, was more severely impacted by low doses of radiation than that of desiccation-resistant species. Surprisingly, the radioresistance of desiccation-resistant species is not related to features of their original habitat. Indeed, bdelloids isolated from Atacama Desert or Antarctica were not characterized by a higher radioresistance than species found in more temperate environments.ConclusionsTolerance to desiccation and radiation are supported as ancestral features of bdelloid rotifers, with a group of species of the genus Rotaria having lost this trait after colonizing permanent water habitats. Together, our results provide a comprehensive overview of the evolution of desiccation and radiation resistance among bdelloid rotifers.

Transgenic expression of late embryogenesis abundant proteins improves tolerance to water stress in Drosophila melanogaster.

Four lines of Drosophila melanogaster were created that expressed transgenes encoding selected late embryogenesis abundant (LEA) proteins originally identified in embryos of the anhydrobiote Artemia franciscana The overall aim was to extend our understanding of the protective properties of LEA proteins documented with isolated cells to a desiccation-sensitive organism during exposure to drying and hyperosmotic stress. Embryos of D. melanogaster were dried at 57% relative humidity to promote a loss of 80% tissue water and then rehydrated. Embryos that expressed AfrLEA2 or AfrLEA3m eclosed 2days earlier than wild-type embryos or embryos expressing green fluorescent protein (Gal4GFP control). For the third instar larval stage, all Afrlea lines and Gal4GFP controls experienced substantial drops in survivorship as desiccation proceeded. When results for all Afrlea lines were combined, Kaplan-Meier survival curves indicated a significant improvement in survivorship in fly lines expressing AfrLEA proteins compared with Gal4GFP controls. The percent water lost at the LT50 (lethal time for 50% mortality) for the AfrLEA lines was 78% versus 52% for Gal4GFP controls. Finally, offspring of fly lines that expressed AfrLEA2, AfrLEA3m or AfrLEA6 exhibited significantly greater success in reaching pupation, compared with wild-type flies, when adults were challenged with hyperosmotic stress (NaCl-fortified medium) and progeny forced to develop under these conditions. In conclusion, the gain of function studies reported here show that LEA proteins can improve tolerance to water stress in a desiccation-sensitive species that normally lacks these proteins, and, simultaneously, underscore the complexity of desiccation tolerance across multiple life stages in multicellular organisms.

Ecological insights into the coexistence of dormancy and desiccation-sensitivity in Arecaceae species

• Key message Mechanisms controlling germination of > 2600 Arecaceae species adapted to the tropical forests are not clearly understood. This contribution underpins the notion that the coexistence of dormancy and desiccation sensitivity in some Arecaceae species could significantly affect the germination ecology. Although the evolutionary and ecological significance behind this occurrence is unknown, these traits together could limit the germination from inappropriate depths and allow dispersal. • Context Dormancy and desiccation sensitivity, i.e., recalcitrance, are considered as mutually exclusive traits that evolved to maximize seed survival after dispersal; the former by allowing seeds to spread germination risk temporally then synchronize germination with a favorable growing season, and the latter by allowing germination immediately. However, a few species from temperate ecosystems have been known to produce seeds that are both dormant and desiccation-sensitive, but little is known about such a relationship in the tropical forests. • Aims To understand if desiccation sensitivity and dormancy can coexist in Arecaceae species, distributed predominantly in the tropical forests, and compare the seed traits found in Arecaceae species with other desiccation-sensitive species. • Methods Information published in international peer-reviewed journals was reviewed and a database with records on seed dormancy and desiccation sensitivity was created and discussed. • Results This literature survey has identified diaspores of 34 Arecaceae species that have both desiccation-sensitivity and an underdeveloped, small embryo, i.e., morphophysiological dormancy or morphological dormancy. In Arecaceae species, desiccation-sensitivity is neither confined to large-sized diaspores, nor all species adapted to dry ecosystems disperse diaspore during the wet-season; features reported to be prevalent in desiccation-sensitive species of other families. Endocarp is proposed to act as a mechanical barrier preventing water loss and also offer protection against physical damage; however, the extent of protection against water loss from internal structures is contentious. • Conclusion The possible ecological significance of this unique relationship noted in Arecaceae is currently unknown, and this review puts forward “testable” hypotheses that call for more studies on germination ecology focusing on dispersal and burial.

Resurrection plants optimize photosynthesis despite very thick cell walls by means of chloroplast distribution

Resurrection plants are vascular species able to sustain extreme desiccation in their vegetative tissues. Despite its potential interest, the role of leaf anatomy in CO2 diffusion and photosynthesis under non-stressed conditions has not been explored in these species. Net CO2 assimilation (An) and its underlying diffusive, biochemical and anatomical determinants were assessed in 10 resurrection species from diverse locations, including ferns and homoiochlorophyllous and poikilochlorophyllous angiosperms. Data obtained were compared to previously published results in desiccation-sensitive ferns and angiosperms. An in resurrection plants was mostly driven by mesophyll conductance to CO2 (gm) and limited by CO2 diffusion. Resurrection species had a larger cell wall thickness (Tcw) than desiccation sensitive plants, a feature associated with limited CO2 diffusion in the mesophyll, but also greater chloroplast exposure to intercellular spaces (Sc), which usually leads to higher gm. This combination enabled a higher An per Tcw compared to desiccation sensitive species. Resurrection species possess unusual anatomical features that possibly confer stress tolerance (thick cell walls) without compromising the photosynthetic capacity (high chloroplast exposure). This mechanism is particularly successful in resurrection ferns, which display higher photosynthesis than their desiccation sensitive counterparts.

Comparative transcriptome analysis suggests convergent evolution of desiccation tolerance in Selaginella species

BackgroundDesiccation tolerant Selaginella species evolved to survive extreme environmental conditions. Studies to determine the mechanisms involved in the acquisition of desiccation tolerance (DT) have focused on only a few Selaginella species. Due to the large diversity in morphology and the wide range of responses to desiccation within the genus, the understanding of the molecular basis of DT in Selaginella species is still limited.ResultsHere we present a reference transcriptome for the desiccation tolerant species S. sellowii and the desiccation sensitive species S. denticulata. The analysis also included transcriptome data for the well-studied S. lepidophylla (desiccation tolerant), in order to identify DT mechanisms that are independent of morphological adaptations. We used a comparative approach to discriminate between DT responses and the common water loss response in Selaginella species. Predicted proteomes show strong homology, but most of the desiccation responsive genes differ between species. Despite such differences, functional analysis revealed that tolerant species with different morphologies employ similar mechanisms to survive desiccation. Significant functions involved in DT and shared by both tolerant species included induction of antioxidant systems, amino acid and secondary metabolism, whereas species-specific responses included cell wall modification and carbohydrate metabolism.ConclusionsReference transcriptomes generated in this work represent a valuable resource to study Selaginella biology and plant evolution in relation to DT. Our results provide evidence of convergent evolution of S. sellowii and S. lepidophylla due to the different gene sets that underwent selection to acquire DT

Profiling of phenolic compounds in desiccation-tolerant and non-desiccation-tolerant Linderniaceae.

Craterostigma plantagineum and Lindernia brevidens are resurrection plants, so these plants can tolerate desiccation of their vegetative tissues. Different components and mechanisms contribute to desiccation tolerance and secondary plant metabolites, like phenolic compounds, may play a role during these processes. Secondary plant metabolites of the two resurrection plants, C. plantagineum and L. brevidens as well as the closely related desiccation sensitive species, L. subracemosa, were investigated regarding the polyphenol profile. Secondary plant compounds were extracted with acidified methanol and analysed with ultra-high-performance liquid chromatography electrospray ionisation mass spectrometry (UHPLC-ESI-MS). Phenolic compounds were identified by comparing of ultraviolet (UV) and MSn -spectra with published data. All compounds were quantified as verbascoside equivalents by external calibration at the compound specific wavelength. In total, eight compounds that belong to the subclass of phenylethanoid glycosides and one flavone, luteolin hexoside pentoside, were identified. Two of these compounds exhibited a fragmentation pattern, which is closely related to phenylethanoid glycosides. The predominantly synthesised phenylethanoid in all of the three plant species and in every stage of hydration was verbascoside. The total content of phenolic compounds during the three stages of hydration, untreated, desiccated, and rehydrated revealed differences especially between C. plantagineum and L. brevidens as the latter one lost almost all phenolic compounds during rehydration. The amount of verbascoside correlates with the degree of desiccation tolerance and verbascoside might play a role in the protective system in acting as an antioxidant.

Identification of a master transcription factor and a regulatory mechanism for desiccation tolerance in the anhydrobiotic cell line Pv11.

Water is essential for living organisms. Terrestrial organisms are incessantly exposed to the stress of losing water, desiccation stress. Avoiding the mortality caused by desiccation stress, many organisms acquired molecular mechanisms to tolerate desiccation. Larvae of the African midge, Polypedilum vanderplanki, and its embryonic cell line Pv11 tolerate desiccation stress by entering an ametabolic state, anhydrobiosis, and return to active life after rehydration. The genes related to desiccation tolerance have been comprehensively analyzed, but transcriptional regulatory mechanisms to induce these genes after desiccation or rehydration remain unclear. Here, we comprehensively analyzed the gene regulatory network in Pv11 cells and compared it with that of Drosophila melanogaster, a desiccation sensitive species. We demonstrated that nuclear transcription factor Y subunit gamma-like, which is important for drought stress tolerance in plants, and its transcriptional regulation of downstream positive feedback loops have a pivotal role in regulating various anhydrobiosis-related genes. This study provides an initial insight into the systemic mechanism of desiccation tolerance.

Impacts of rainfall extremes predicted by climate-change models on major trophic groups in the leaf litter arthropod community.

Arthropods in the leaf litter layer of forest soils influence ecosystem processes such as decomposition. Climate-change models predict both increases and decreases in average rainfall. Increased drought may have greater impacts on the litter arthropod community. In addition to affecting survival or behaviour of desiccation-sensitive species, lower rainfall may indirectly lower abundances of consumers that graze drought-stressed fungi, with repercussions for higher trophic levels. We tested the hypothesis that trophic structure will differ between the two rainfall scenarios. In particular, we hypothesized that densities of several broadly defined trophic groupings of arthropods would be lower under reduced rainfall. To test this hypothesis, we used sprinklers to impose two rainfall treatments during three growing seasons in roofed, fenced 14-m2 plots and documented changes in abundance from initial, pre-treatment densities of 39 arthropod taxa. Experimental plots were subjected to either LOW (fortnightly) or HIGH (weekly) average rainfall based upon climate models and the previous 100years of regional weekly averages. Unroofed open plots, our reference treatment (REF), experienced higher than average rainfall during the experiment. The two rainfall extremes produced clear negative effects of lowered rainfall on major trophic groups. Broad categories of fungivores, detritivores and predators were more abundant in HIGH than LOW plots by the final year. Springtails (Collembola), which graze fungal hyphae, were 3× more abundant in the HIGH rainfall treatment. Taxa of larger-bodied fungivores and detritivores, spiders (Araneae), and non-spider predators were 2× more abundant under HIGH rainfall. Densities of mites (Acari), which include fungivores, detritivores and predators, were 1.5× greater in HIGH rainfall plots. Abundances and community structure of arthropods were similar in REF and experimental plots, showing that effects of rainfall uncovered in the experiment are applicable to nature. This pattern suggests that changes in rainfall will alter bottom-up control processes in a critical detritus-based food web of deciduous forests. Our results, in conjunction with other findings on the impact of desiccation on arthropods and fungal growth, suggest that drier conditions will depress densities of fungal consumers, causing declines in higher trophic levels, with possible impacts on soil processes and the larger forest food web.

Massive Tandem Proliferation of ELIPs Supports Convergent Evolution of Desiccation Tolerance across Land Plants.

Desiccation tolerance was a critical adaptation for the colonization of land by early nonvascular plants. Resurrection plants have maintained or rewired these ancestral protective mechanisms, and desiccation-tolerant species are dispersed across the land plant phylogeny. Although common physiological, biochemical, and molecular signatures are observed across resurrection plant lineages, features underlying the recurrent evolution of desiccation tolerance are unknown. Here we used a comparative approach to identify patterns of genome evolution and gene duplication associated with desiccation tolerance. We identified a single gene family with dramatic expansion in all sequenced resurrection plant genomes and no expansion in desiccation-sensitive species. This gene family of early light-induced proteins (ELIPs) expanded in resurrection plants convergent through repeated tandem gene duplication. ELIPs are universally highly expressed during desiccation in all surveyed resurrection plants and may play a role in protecting against photooxidative damage of the photosynthetic apparatus during prolonged dehydration. Photosynthesis is particularly sensitive to dehydration, and the increased abundance of ELIPs may help facilitate the rapid recovery observed for most resurrection plants. Together, these observations support convergent evolution of desiccation tolerance in land plants through tandem gene duplication.

Desiccation tolerance in plants: Structural characterization of the cell wall hemicellulosic polysaccharides in three Selaginella species

Drought-induced dehydration of vegetative tissues in lycopods affects growth and survival. Different species of Selaginella have evolved a series of specialized mechanisms to tolerate desiccation in vegetative tissues in response to water stress. In the present study, we report on the structural characterization of the leaf cell wall of the desiccation-tolerant species S. involvens and two desiccation-sensitive species, namely S. kraussiana and S. moellendorffii. Isolated cell walls from hydrated and desiccated leaves of each species were fractionated and the resulting oligosaccharide fragments were analyzed to determine their structural features. Our results demonstrate that desiccation induces substantial modifications in the cell wall composition and structure. Altogether, these data highlight the fact that structural remodeling of cell wall hemicellulosic polysaccharides including XXXG-rich xyloglucan, arabinoxylan and acetylated galactomannan is an important process in order to mitigate desiccation stress in Selaginella.

Analysis of pcC13-62 promoters predicts a link between cis-element variations and desiccation tolerance in Linderniaceae.

Reproductive structures of plants (e.g. seeds) and vegetative tissues of resurrection plants can tolerate desiccation. Many genes encoding desiccation-related proteins (DRPs) have been identified in the resurrection plant Craterostigma plantagineum, but the function of these genes remains mainly hypothetical. Here, the importance of the DRP gene pcC13-62 for desiccation tolerance is evaluated by analysing its expression in C. plantagineum and in the closely related desiccation-tolerant species Lindernia brevidens and the desiccation-sensitive species Lindernia subracemosa. Quantitative analysis revealed that pcC13-62 transcripts accumulate at a much lower level in desiccation-sensitive species than in desiccation-tolerant species. The study of pcC13-62 promoters from these species demonstrated a correlation between promoter activity and gene expression levels, suggesting transcriptional regulation of gene expression. Comparison of promoter sequences identified a dehydration-responsive element motif in the promoters of tolerant species that is required for dehydration-induced β-glucuronidase (GUS) accumulation. We hypothesize that variations in the regulatory sequences of the pcC13-62 gene occurred to establish pcC13-62 expression in vegetative tissues, which might be required for desiccation tolerance. The pcC13-62 promoters could also be activated by salt stress in Arabidopsis thaliana plants stably transformed with promoter::GUS constructs.

Zygotic embryo cell wall responses to drying in three gymnosperm species differing in seed desiccation sensitivity.

Plant cell walls (CWs) are dynamic in that they can change conformation during ontogeny and in response to various stresses. Though seeds are the main propagatory units of higher plants, little is known of the conformational responses of zygotic embryo CWs to drying. This study employed cryo-scanning electron microscopy to compare the effects of desiccation on zygotic embryo CW morphology across three gymnosperm species that were shown here to differ in seed desiccation sensitivity: Podocarpus henkelii (highly desiccation-sensitive), Podocarpus falcatus (moderately desiccation-sensitive), and Pinus elliottii (desiccation-tolerant). Fresh/imbibed (i.e. fresh Podocarpus at shedding and imbibed Pi. elliottii) embryos showed polyhedral cells with regular walls, typical of turgid cells with an intact plasmalemma. Upon desiccation to c. 0.05gg-1 (dry mass basis), CWs assumed an undulating conformation, the severity of which appeared to depend on the amount and type of dry matter accumulated. After desiccation, intercellular spaces between cortical cells in all species were comparably enlarged relative to those of fresh/imbibed embryos. After rehydration, meristematic and cotyledonary CWs of P. henkelii and meristematic CWs of P. falcatus remained slightly undulated, suggestive of plasmalemma and/or CW damage, while those of Pi. elliottii returned to their original conformation. Cell areas in dried-rehydrated P. henkelii root meristem and cotyledon were also significantly lower than those from fresh embryos, suggesting incomplete recovery, even though embryo water contents were comparable between the two states. Electrolyte leakage measurements suggest that the two desiccation-sensitive species incurred significant plasmalemma damage relative to the tolerant species upon desiccation, in agreement with the CW abnormalities observed in these species after rehydration. Immunocytochemistry studies revealed that of the four CW epitopes common to embryos of all three species, an increase in arabinan (LM6) upon desiccation and rehydration in desiccation-tolerant Pi. elliottii was the only difference, although this was not statistically significant. Seed desiccation sensitivity in species like P. henkelii and P. falcatus may therefore be partly based on the inability of the plasmalemma and consequently CWs of dried embryos to regain their original conformation following rehydration.

Taxonomic affinity, habitat and seed mass strongly predict seed desiccation response: a boosted regression trees analysis based on 17 539 species.

Seed desiccation response plays an important role in plant regeneration ecology, and has significant implications for species conservation. The majority of seed plants produce desiccation-tolerant (orthodox) seeds, whilst comparatively few produce desiccation-sensitive (recalcitrant) seeds that are unable to survive dehydration, and which cannot be conserved in traditional seed banks. This study develops a set of models to predict seed desiccation response in unstudied species. Taxonomy, trait, location and climate data were compiled to form a global data set of 17 539 species. Three boosted regression trees models were then developed to predict species' seed desiccation responses based on habitat and trait information for the species, and the seed desiccation responses of close relatives (either members of the same genus, family or order, depending on the model). Ten-fold cross-validation was used to test model predictive success. The utility of the models was then demonstrated by predicting seed desiccation response for two floras: Ecuador, and Britain and Ireland. The three models had varying success rates for identifying the desiccation-sensitive species: 89 % for the genus-level model, 79 % for the family-level model and 60 % for the order-level model. The most important predictor variables were the seed desiccation responses of a species' relatives, seed mass and annual precipitation. It is predicted that 10 % of seed plants from Ecuador and 1.2 % of those from Britain and Ireland produce desiccation-sensitive seeds. Due to data availability, prediction accuracy is likely to be higher for the British and Irish flora, where it is estimated that a desiccation-sensitive species had a 96.7 % chance of being correctly identified, compared with 80.8 % in the Ecuador flora. These models can utilize existing data to predict species' likely seed desiccation responses, providing a gap-filling tool for global studies of plant traits, as well as critical decision-making support for plant conservation activities.

Glycerolipid analysis during desiccation and recovery of the resurrection plant Xerophyta humilis (Bak) Dur and Schinz.

Xerophyta humilis is a poikilochlorophyllous monocot resurrection plant used as a model to study vegetative desiccation tolerance. Dehydration imposes tension and ultimate loss of integrity of membranes in desiccation sensitive species. We investigated the predominant molecular species of glycerolipids present in root and leaf tissues, using multiple reaction monitoring mass spectrometry, and then analysed changes therein during dehydration and subsequent rehydration of whole plants. The presence of fatty acids with long carbon chains and with odd numbers of carbons were detected and confirmed by gas chromatography. Dehydration of both leaves and roots resulted in an increase in species containing polyunsaturated fatty acids and a decrease in disaturated species. Upon rehydration, lipid saturation was reversed, with this being initiated immediately upon watering in roots but only 12-24hr later in leaves. Relative levels of species with short-chained odd-numbered saturated fatty acids decreased during dehydration and increased during rehydration, whereas the reverse trend was observed for long-chained fatty acids. X.humilis has a unique lipid composition, this report being one of the few to demonstrate the presence of odd-numbered fatty acids in plant phosphoglycerolipids.

Expression profile of desiccation tolerance factors in intertidal seaweed species during the tidal cycle.

The transcriptional modulation of desiccation tolerance factors in P. orbicularis explains its successful recuperation after water deficit. Differential responses to air exposure clarify seaweed distribution along intertidal rocky zones. Desiccation-tolerant seaweed species, such as Pyropia orbicularis, can tolerate near 96% water loss during air exposure. To understand the phenotypic plasticity of P. orbicularis to desiccation, several tolerance factors were assessed by RT-qPCR, Western-blot analysis, and enzymatic assays during the natural desiccation-rehydration cycle. Comparative enzymatic analyses were used to evidence differential responses between P. orbicularis and desiccation-sensitive species. The results showed that during desiccation, the relative mRNA levels of genes associated with basal metabolism [trehalose phosphate synthase (tps) and pyruvate dehydrogenase (pdh)] were overexpressed in P. orbicularis. Transcript levels related to antioxidant metabolism [peroxiredoxin (prx); thioredoxin (trx); catalase (cat); lipoxygenase (lox); ferredoxin (fnr); glutathione S-transferase (gst)], cellular detoxification [ABC transporter (abc) and ubiquitin (ubq)], and signal transduction [calmodulin (cam)] increased approximately 15- to 20-fold, with the majority returning to basal levels during the final hours of rehydration. In contrast, actin (act) and transcription factor 1 (tf1) transcripts were down-regulated. ABC transporter protein levels increased in P. orbicularis during desiccation, whereas PRX transcripts decreased. The antioxidant enzymes showed higher specific activity in P. orbicularis under desiccation, and sensitive species exhibited enzymatic inactivation and scarce ABC and PRX protein detection following prolonged desiccation. In conclusion, the reported findings contribute towards understanding the ecological distribution of intertidal seaweeds at the molecular and functional levels.

A New Physiological Role for the DNA Molecule as a Protector against Drying Stress in Desiccation-Tolerant Microorganisms.

The DNA molecule is associated with the role of encoding information required to produce RNA which is translated into proteins needed by the cell. This encoding involves information transmission to offspring or to other organisms by horizontal transfer. However, despite the abundance of this molecule in both the cell and the environment, its physiological role seems to be restricted mainly to that of a coding and inheritance molecule. In this paper, we report a new physiological role for the DNA molecule as involved in protection against desiccation, in addition to its well-established main information transfer and other recently reported functions such as bio-film formation in eDNA form. Desiccation-tolerant microorganisms such as Microbacterium sp. 3J1 significantly upregulate genes involved in DNA synthesis to produce DNA as part of their defensive mechanisms to protect protein structures and functions from drying according to RNA-seq analysis. We have observed the intracellular overproduction of DNA in two desiccation-tolerant microorganisms, Microbacterium sp. 3J1 and Arthrobacter siccitolerans 4J27, in response to desiccation signals. In addition, this conclusion can be made from our observations that synthetic DNA protects two proteins from drying and when part of a xeroprotectant preparation, DNA from various organisms including desiccation-sensitive species, does the same. Removal of DNA by nuclease treatment results in absence of this additive protective effect. We validated this role in biochemical and biophysical assays in proteins and occurs in trans even with short, single chains of synthetically produced DNA.

Establishment of an isogenic strain of the desiccation-sensitive tardigradeIsohypsibius myrops(Parachela, Eutardigrada) and its life history traits

Tardigrades are aquatic micrometazoans found in various habitats. Many limno-terrestrial species can tolerate desiccation by entering a metabolically inactive dehydrated state called anhydrobiosis, whereas most species in constantly hydrated habitats, such as freshwater or marine species, cannot undergo anhydrobiosis and are susceptible to desiccation. Although comparative analyses between anhydrobiotic species and desiccation-sensitive species will lead to a better understanding of anhydrobiosis, almost all studies on anhydrobiosis have been performed only in anhydrobiotic species. In the present study, we established a rearing system and an isogenic strain of Isohypsibius myrops collected from activated sludge. This strain is highly sensitive to desiccation and cannot tolerate exposure to relative humidity (RH) of 97–98% for 2 days or 94–95% for 1 day. Life history traits revealed that this strain has a short hatching time (3.6 days) and a short lifespan (18.8 days). The strain frequently lays eggs, on average 19 eggs per oviposition at mean interval of 2.5 days, enabling their rapid proliferation. The high fecundity of desiccation-sensitive species implies a possible trade-off between anhydrobiotic ability and fecundity. The established strain provides a good experimental model of desiccation-sensitive tardigrades, especially for comparative molecular analyses of anhydrobiosis mechanisms.

LEA gene expression, RNA stability and pigment accumulation in three closely related Linderniaceae species differing in desiccation tolerance

Desiccation-tolerant plants (Craterostigma plantagineum and Lindernia brevidens) evolved a highly efficient strategies to prevent dehydration-induced irreversible damage. The protection system involves synthesis of LEA proteins, decrease of photosynthetic activity and activation of antioxidant systems. The regulation of these processes requires joint action of multiple proteins. Here, we present comparative analyses of accumulation of transcripts encoding components of the protection machinery, such as selected LEA proteins, enzymes of the chlorophyll degradation pathway and anthocyanin biosynthesis enzymes in total and polysomal RNA pools. The analyses revealed that desiccation-tolerant plants recruit mRNAs to ribosomes with higher efficiency than the desiccation-sensitive species L. subracemosa. Desiccation-tolerant species accumulated high amounts of LEA transcripts during dehydration and precisely controlled the amounts of chlorophyll keeping it at a level sufficient to activate photosynthesis after rehydration. In contrast, mRNA of L. subracemosa was prone to dehydration-induced degradation, decomposition of the photosynthetic apparatus and degradation of free chlorophyll. Thus, the results of the studies point to differences in the control of gene expression and degradation of chlorophyll in desiccation-tolerant versus desiccation-sensitive species when the plants were subjected to dehydration.