Sporophyte and gametophyte generations differ in their thermotolerance response in the moss Microbryum

BACKGROUND AND AIMS: Actively growing post-embryonic sporophytes of desert mosses are restricted to the cooler, wetter months. However, most desert mosses have perennial gametophytes. It is hypothesized that these life history patterns are due in part to a reduced thermotolerance for sporophytes relative to gametophytes. METHODS: Gametophytes with attached embryonic sporophytes of Microbryum starckeanum were exposed whilst desiccated to thermal episodes of 35 degrees C (1 hr), 55 degrees C (1 hr), 75 degrees C (1 hr) and 75 degrees C (3 hr), then moistened and allowed to recover for 35 d in a growth chamber. KEY RESULTS: All of the gametophytes survived the thermal exposures and produced protonemata, with the majority also producing shoot buds. Symptoms of gametophytic stress (leaf burning and discoloration of entire shoots) were present in lower frequencies in the 55 degrees C exposure. Sporophyte resumption of growth and maturation to meiosis were significantly negatively affected by thermal treatment. Not a single sporophyte exposed to the two higher thermal treatments (75 degrees C for 1 h and 75 degrees C for 3 h) survived to meiosis, and those sporophytes exposed to 75 degrees C that survived to the post-embryonic phenophase took significantly longer to reach this phase. Furthermore, among the thermal treatments where some capsules reached maturity (35 degrees C and 55 degrees C), maternal shoots that produced a meiotic capsule took longer to regenerate through protonemata than maternal shoots aborting their sporophyte, suggestive of a resource trade-off between generations. CONCLUSIONS: Either (1) the inherent sporophyte thermotolerance is quite low even in this desert moss, and/or (2) a gametophytic thermal stress response controls sporophyte viability.     

Developing sporophytes transition from an inducible to a constitutive ecological strategy of desiccation tolerance in the moss Aloina ambigua: effects of desiccation on fitness

Background and Aims Two ecological strategies of desiccation tolerance exist in plants, constitutive and inducible. Because of difficulties in culturing sporophytes, very little is known about desiccation tolerance in this generation and how desiccation affects sexual fitness.Methods Cultured sporophytes and vegetative shoots from a single genotype of the moss Aloina ambigua raised in the laboratory were tested for their strategy of desiccation tolerance by desiccating the shoot–sporophyte complex and vegetative shoots at different intensities, and comparing outcomes with those of undried shoot–sporophyte complexes and vegetative shoots. By using a dehardened clonal line, the effects of field, age and genetic variance among plants were removed.Key Results The gametophyte and embryonic sporophyte were found to employ a predominantly inducible strategy of desiccation tolerance, while the post-embryonic sporophyte was found to employ a moderately constitutive strategy of desiccation tolerance. Further, desiccation reduced sporophyte fitness, as measured by sporophyte mass, seta length and capsule size. However, the effects of desiccation on sporophyte fitness were reduced if the stress occurred during embryonic development as opposed to postembryonic desiccation.Conclusions The effects of desiccation on dehardened sporophytes of a bryophyte are shown for the first time. The transition from one desiccation tolerance strategy to the other in a single structure or generation is shown for only the second time in plants and for the first time in bryophytes. Finding degrees of inducible strategies of desiccation tolerance in different life phases prompts the formulation of a continuum hypothesis of ecological desiccation tolerance in mosses, where desiccation tolerance is not an either/or phenomenon, but varies in degree along a gradient of ecological inducibility.     

Morphological adaptation to water uptake and transport in the poikilohydric moss Tortula ruralis

Abstract

In this paper different water-holding and transport adaptations to face the uneven and intermittent distribution of water in the ectohydric moss Tortula ruralis are referred to.

The external conduction of water is operated by different and efficient systems (spaces between adjacent shoots, between leaves, leaves and stems, leaves and rhizoids, life form, etc.) which facilitate absorption and transport of solutions.

Also epi-organ capillary systems (bases and revoluted margin of leaves, a groove, a close network of capillary channels determined by the papillae), intra-organ capillary systems (hyalocysts) and certain amount of internal conduction (conducting parenchyma, stereoma) co-operante to absorb and distribute water to the whole gametophyte.

In addition, some adaptations to xerophytism and heliophytism (life form, disposition of phylloids, hair-points) are discussed.     

Evidence of stress imprinting with population‐level differences in two moss species

Plants are often repeatedly exposed to stresses during their lives and have a mechanism called stress imprinting that provides “memories” of stresses they experience and increases their ability to cope with later stresses. To test hypotheses that primed bryophytes can preserve their stress imprinting after 6 days of recovery and induce higher levels of osmolytes and ROS‐scavenging activities upon later stress exposure, and there exist population‐level differentiation in their desiccation defenses, we transplanted samples of two populations of each of two moss species, Hypnum plumaeforme and Pogonatum cirratum, in a nature reserve in southern China. After 16 months of acclimation, sets of each population were subjected to control, one‐time desiccation stress, duplicated desiccation stress and cross‐stress (low temperature stress followed by desiccation stress) treatments. Levels of oxidant enzymes, osmolytes, and phytohormones in the samples were then determined. The desiccation stress generally led to increases in activities or contents of superoxide dismutase, guaiacol peroxidase, catalase, proline, soluble sugars, soluble proteins, and stress hormones including abscisic acid (ABA), jasmonates (JA), and salicylic acid (SA), with differences between both species and populations. After a 6‐day recovery period, contents of phytohormones (including ABA, JA, SA, and cytokinins) in stressed H. plumaeforme had substantially fallen toward control levels. The duplicated and cross‐stress treatments generally led to further accumulation of proline, soluble sugars, and soluble proteins, with further increases in activities of antioxidant enzymes in some cases. Furthermore, significant differences between allochthonous and native populations were found in contents of malondialdehyde and osmolytes, as well as antioxidant enzyme activities. Our results confirm the hypotheses and highlight the importance of osmolytes in mosses' stress responses.     

Physiological history may mask the inherent inducible desiccation tolerance strategy of the desert moss Crossidium crassinerve

Shoots of bryophytes collected in the desiccated state from the field are likely to be hardened to desiccation tolerance (DT) to varying degrees. To account for this, most studies on DT include a relatively short deacclimation period. However, no study has experimentally determined the appropriate deacclimation time for any bryophyte species. Our purposes are to (i) determine if ‘field effects’ are biologically relevant to DT studies and how long a deacclimation period is required to remove them; and (ii) utilise field versus cultured shoot responses within the context of a deacclimation period to elucidate the ecological strategy of DT. Our hypothesis (based on an extensive literature on DT) is that a deacclimation period from 24 to 72 h should be sufficient to eliminate historical stress effects on the physiology of the shoots and allow an accurate determination of the inherent ecological DT strategy (constitutive or inducible). We determined, however, using chlorophyll fluorescence and visual estimates of shoot damage, that field‐collected shoots of the desert moss Crossidium crassinerve required an experimental deacclimation period of >7 days before field effects were removed, and revealed an ecological DT strategy of inducible DT. If the deacclimation period was <6 days, the shoot response conformed to an ecological strategy of constitutive protection. Thus the presence of field effects can obscure the ecological strategy of desiccation tolerance exhibited by the species, and this translates into a need to re‐evaluate previous mechanistic and ecological studies of desiccation tolerance in plants.     

Gender differences and a new adult eukaryotic record for upper thermal tolerance in the desert moss Syntrichia caninervis

1.

Two competing hypotheses relating to thermostress were proposed to understand skewed sex ratios in Syntrichia caninervis, a reproductive investment hypothesis and a wildfire selection hypothesis.     

Diurnal variation in photochemical dynamics and surface reflectance of the desiccation-tolerant moss,Tortula ruralis

Most desiccation-tolerant plants alter shoot structure during drying, making it possible to use changes in surface reflectance as a proxy measure of plant water status. Diurnal courses of surface reflectance (albedo) and chlorophyll fluorescence parameters of the ectohydric moss, Tortula ruralis (Hedw.) Gaertn, were measured to assess the coordination between anatomical and physiological features under field conditions. Albedo showed a sigmoidal relationship with relative humidity and the deviation of moss mat temperature from dew point. Maximum photosynthetic quantum yield (F _v/F _m) also displayed a sigmoidal relationship pooled across three days differing in light, temperature, and relative humidity. Depending on the light conditions and rapidity of drying during the morning, there were distinct differences in the ability of T. ruralis to establish thermal dissipation of excess light energy (NPQ) across a range of light levels following rehydration through the day. These findings suggest that there is a coordinated suite of architectural and physiological characteristics maintaining the photosynthetic integrity of these plants in highly variable arid and semi-arid environments.     

Bryophytes as experimental models for the study of environmental stress tolerance: Tortula ruralis and desiccation-tolerance in mosses

The development of a complete understanding of how plants interact with the environment at the cellular level is a crucial step in advancing our ability to unravel the complexities of plant ecology particularly with regard to the role that many of the less complex plants (i.e., algae, lichens, and bryophytes) play in plant communities and in establishing areas for colonization by their more complex brothers. One of the main barriers to the advancement of this area of plant biology has been the paucity of simple and appropriate experimental models that would enable the researcher to biochemically and genetically dissect the response of less complex plants to environmental stress. A number of bryophytes model systems have been developed and they have been powerful experimental tools for the elucidation of complex biological processes in plants. Recently there has been a resurgent interest in bryophytes as models systems due to the discovery and development of homologous recombination technologies in the moss Physcomitrella patens (Hedw.) Brach & Schimp. In this report we introduce the desiccation-tolerant moss Tortula ruralis (Hedw.) Gaert., Meyer, and Scherb, as a model for stress tolerance mechanisms that offers a great deal of promise for advancing our efforts to understand how plants respond to and survive the severest of stressful environments. T. ruralis, a species native to Northern and Western North America, has been the most intensely studied of all bryophytes with respect to its physiological, biochemical, and cellular responses, to the severest of water stresses, desiccation. It is our hope that the research conducted using this bryophyte will lay the foundationfor not only the ecology of bryophytes and other less complex plants but also for the role of desiccation-tolerance in the evolution of land plants and the determination of mechanisms by which plant cells can withstand environmental insults. We will focus the discussion on the research we and others have conducted in an effort to understand the ability of T. ruralis to withstand the complete loss of free water from the protoplasm of its cells.     

Exaggeration of desiccation stress by heavy metal pollution in Tortula ruralis: a pilot study

Glutathione pool and redox status, as well as chlorophyll fluorescencewere measured in Tortula ruralis which was treated withheavy metals and exposed to different desiccation treatments. Two hours afterre-wetting, the ratio of oxidised glutathione to total glutathione poolreturnedto the steady state level (14%) in slowly dried unpolluted plants. Cdtreatment doubled this ratio, as did rapid drying without the heavy metaltreatment. When Cd and rapid drying were applied together, the ratio of GSSGreached 45% indicating a clear additive effect of these two stressfactors. RFd, a chlorophyll fluorescence parameter followed a similar pattern.Lead did not cause the depletion of the glutathione pool but increased theratioof GSSG. It is suggested that Cd and rapid desiccation exert their damageadditively. This might also entail a lowered degree of desiccation tolerance inareas polluted with metals and therefore a retreat of the mosses to mesicmicrohabitats.     

Ecological and evolutionary consequences of desiccation tolerance in tropical fern gametophytes

Ferns have radiated into the same diverse environments as spermatophytes, and have done so with an independent gametophyte that is not protected by the parent plant. The degree and extent of desiccation tolerance (DT) in the gametophytes of tropical fern species was assessed to understand mechanisms that have allowed ferns to radiate into a diversity of habitats. Species from several functional groups were subjected to a series of desiccation events, including varying degrees of intensity and multiple desiccation cycles. Measurements of chlorophyll fluorescence were used to assess recovery ability and compared with species ecology and gametophyte morphology. It is shown that vegetative DT (rare in vascular plants) is widely exhibited in fern gametophytes and the degree of tolerance is linked to species habitat preference. It is proposed that gametophyte morphology influences water-holding capacity, a novel mechanism that may help to explain how ferns have radiated into drought-prone habitats. Fern gametophytes have often been portrayed as extreme mesophytes with little tolerance for desiccation. The discovery of DT in gametophytes holds potential for improving our understanding of both the controls on fern species distribution and their evolution. It also advances a new system with which to study the evolution of DT in vascular plants.     

Sex expression and sex dimorphism in sporophytic populations of the desert moss Syntrichia caninervis

Ten sporophytic populations of Syntrichiacaninervis contained an average of 22 individualscm^–2, with a sex ratio of 7.9 Female:1 Male: 3.1 Nonexpressing(N = 300). In each of the populations, female individuals outnumberedmale individuals. A representative population size of 50 × 25cm was estimated to contain 27,250 organically distinct individualplants. Given the wide disparity in reproductive investment in this dioeciousspecies, sex-specific traits were investigated at the level of the individualinbiomass, total stem length, number of ramets, number of branches, length ofannual growth interval, length of longest ramet, age, number of inflorescences,number of ramets expressing sex, and consecutive seasons of sex expression. Theonly significant sex dimorphism recorded was in consecutive seasons of sexexpression, with nonsporophytic female individuals exhibiting a higherfrequencythan males. However, nonexpressing individuals had lower biomass, shorter totalstem length, fewer branches, and shorter ramets than males and females, andfewer ramets than female individuals. When the biomass of female and maleindividuals was controlled for inflorescence number, no significant differencesbetween the sexes in biomass were found. There appears to be a threshold sizefor sex expression, with all individuals above 2.0 mg dry weightexpressing sex (N = 108). Biomass and total stem length are strongpositive correlates (r = 0.88), and individual biomass is a betterpredictor of the number of inflorescences produced per individual than is stemlength (r = 0.85 vs. r = 0.69, respectively). Independent of stemlength, individual biomass had a positive and significant relationship withinflorescence number; however, independent of individual biomass, stem lengthwas not associated with inflorescence number. Overall sex expression was 0.74(individual level) and 0.65 (ramet level). The functional sex ratio wasassessedat the inflorescence level, and ranged from 9.7:1, to14.9:1 over the most recent four years. Fertilization frequencywas 0.69 (individual level) and 0.34 (perichaetial level). Over the last threeyears, 63% of all fertilized perichaetia resulted in an abortivesporophyte.     

The use of plant stress biomarkers in assessing the effects of desiccation in zygotic embryos from recalcitrant seeds: challenges and considerations

Zygotic embryos from recalcitrant seeds are sensitive to desiccation. In spite of their sensitivity, rapid partial dehydration is necessary for their successful cryopreservation. However, dehydration to water contents (WCs) that preclude lethal ice crystal formation during cooling and rewarming generally leads to desiccation damage. This study investigated the effects of rapid dehydration on selected stress biomarkers (electrolyte leakage, respiratory competence, rate of protein synthesis, superoxide production, lipid peroxidation, antioxidant activity and degree of cellular vacuolation) in zygotic embryos of four recalcitrant‐seeded species. Most biomarkers indicated differences in the levels of stress/damage incurred by embryos dried to WCs < and >0.4 g·g^?1, within species; however, these changes were often unrelated to viability and percentage water loss when data for the four species were pooled for regression analyses. Dehydration‐induced electrolyte leakage was, however, positively related with percentage water loss, while biomarkers of cellular vacuolation were positively related with both percentage water loss and viability. This suggests that electrolyte leakage and degree of cellular vacuolation can be used to quantify dehydration‐induced stress/damage. Biomarkers such as superoxide production, whilst useful in establishing the nature of the dehydration stress incurred may not be able to distinguish the effects of different WCs/drying times. Irrespective of which biomarker is used, the data suggest that understanding differences in desiccation sensitivity across recalcitrant‐seeded species will remain a challenge unless these biomarkers are related to a generic desiccation stress index that integrates the effects of percentage water loss and drying time.     

Analysis of the decrease in photosynthesis on desiccation of mosses from xeric and hydric environments

Three moss species [ Tortula ruraliformis (Besch.) Grout. Bryum pseudotriquetrum (Hedw.) Schaegr and Dicranella palustris (Dicks.) Crund. ex. E. F. Warb. ( D. squarrosa (Starke) Schp.] collected from a range of habitats differing in water availability were desiccated in controlled conditions. All species became photosynthetically inactive when dried below a water content of 100–200% dry weight. Only Tortula ruraliformis , a moss from arid sand dunes. was able to recover fully to pre-desiccated rates of photosynthetic electron transport during subsequent rehydration. The rate of recovery was influenced by irradiance during desiccation. Mosses from hydric habitats showed some resumption of photosynthetic electron transport (following rehydration) if dried in the dark. but did not do so if dried even in low light. In these circumstances the mosses showed evidence of lasting photoinhibition of photosynthesis after rehydration. The desiccation-tolerant T. ruraliformis became significantly photoinhibited only when continually exposed to high irradiance (1200 μmol m⁻² s⁻¹) in the hydrated state. If allowed to desiccate whilst exposed to high irradiance this species showed less evidence of photoinhibition after rehydration, and was not at all affected by desiccation in low irradiance. Photon flux absorption in dry moss was 50–60% less than that in hydrated moss as a result of leaf curling. However, the reduction in absorption of photosynthetically active radiation cannot account for the total loss of photosynthetic oxygen evolution and variable chlorophyll fluorescence observed in the desiccated mosses.