Desiccoplast-etioplast-chloroplast transformation under rehydration of desiccated poikilochlorophyllous Xerophyta humilis leaves in the dark and upon subsequent illumination.

The transformation of desiccoplasts into etioplasts and the parallel appearance of protochlorophyllide (Pchlide) forms were observed with transmission electron microscopy and 77K fluorescence spectroscopy, when air-dried detached leaves of the poikilochlorophyllous desiccation tolerant plant Xerophyta humilis were floated in water in the dark. After 1 week of rehydration, pregranal plastids with newly synthesized prothylakoid (PT) lamellae and mainly non-photoactive Pchlide forms developed, while etioplasts with prolamellar bodies (PLBs) and photoactive, 655nm emitting Pchlide form accumulated primarily in the basal leaf regions after 2 weeks of regeneration. When these latter leaves were illuminated with continuous light for 3 days, the etioplasts transformed into regular chloroplasts and the fluorescence emission bands characteristic of green leaves appeared. These results show that, upon rehydration, the dehydrated chlorenchyma cells are able to regenerate pregranal plastids and etioplasts from desiccoplasts in the dark, which can transform into regular chloroplasts when they are illuminated. This means that the differentiation of pregranal plastids and etioplasts and their greening process is a basic property of fully differentiated cells of X. humilis. Consequently, these processes are not merely characteristic for seedlings with meristematic and differentiating young tissues.

Winter photosynthetic activity of twenty temperate semi-desert sand grassland species.

The winter photosynthetic activity (quantified by net CO(2) assimilation rates and chlorophyll (Chl) a fluorescence parameters) of 20 plant species (including two lichens and two mosses) of a Hungarian temperate semi-desert sand grassland was determined on one occasion per year in 1984, 1989 and 1994. Throughout winter, the overwintering green shoots, leaves or thalli were regularly exposed to below zero temperatures at night and daytime temperatures of 0-5 degrees C. In situ tissue temperature varied between -2.1 and +6.9 degrees C and the photosynthetic photon flux density (PPFD) between 137 and 351 micromol m(-2)s(-1). Under these conditions 18 of the grassland species exhibited photosynthetic CO(2) uptake (range: vascular plants ca. 0.2-3.8 micromol m(-2)s(-1), cryptogams 0.3-2.79 micromol kg(-1)s(-1)) and values of 0.9-5.1 of the Chl fluorescence decrease ratio R(Fd). In 1984, Festuca vaginata and Sedum sexangulare had net CO(2) assimilation at leaf temperatures of -0.85 to -1.2 degrees C. In 1989, all species except Cladonia furcata showed net CO(2) assimilation at tissue temperatures of 0 to +3.3 degrees C, with the highest rates observed in Poa bulbosa and F. vaginata. The latter showed a net CO(2) assimilation saturation at a PPFD of 600 micromol m(-2)s(-1) and a temperature optimum between +5 and +18 degrees C. At the 1994 measurements, the photosynthetic rates were higher at higher tissue water contents. The two mosses and lichens had a net photosynthesis (range: 1.1-2.79 micromol CO(2)kg(-1)s(-1)) at 2 degrees C tissue temperature and at 4-5 degrees C air temperature. Ca. 80% of the vascular grassland plant species maintained a positive C-balance during the coldest periods of winter, with photosynthetic rates of 1.5-3.8 micromol CO(2)m(-2)s(-1). In an extremely warm beginning March of the relatively warm winter of 2006/2007, the dicotyledonous plants had much higher CO(2) assimilation rates on a Chl (range 6-14.9 micromol g(-1)Chl s(-1)) and on a dry weight basis (9-48 micromol kg(-1)dw s(-1)) than in the cold winter of 1994. However, the assimilation rates of the three investigated cryptogams (Tortula and two Cladonia) and the two grasses Festuca and Poa were not affected by this increase. The results indicate that the photosynthetic activity of temperate semi-desert sand grassland species can help somewhat in slowing the general CO(2) rise in winter and function as a potential carbon sink of the investigated semi-desert Hungarian grassland species.

Tolerance of the lichen Xanthoria parietina (L.) Th. Fr. to metal stress.

Comparisons were made between the content of Al, Cd, Cu, Fe, Mn, Pb, Sb and Zn in thalli of the lichen Xanthoria parietina and selected physiological processes. Assimilation pigments, activity of photosystem II, parietin, reactive substances produced using thiobarbituric acid (TBA) and CO(2) gas exchange were measured in order to assess tolerance to atmospheric sources of pollution. As expected, lichen thalli accumulated high amounts of the measured elements in relation to distance from pollution sources in Kosice, Slovak Republic (US Steel factory and vehicular traffic in the city center). However, except for TBA reactive substances production and to some extent CO(2) gas exchange at the most polluted station, none of the tested physiological parameters showed a clear correlation between accumulation of elements and physiological damage. This reflected a high degree of pollution tolerance in this lichen and corresponded with its high abundance in Kosice.

Long-term acclimation of plants to elevated CO2 and its interaction with stresses.

Rising atmospheric carbon dioxide (CO(2)) concentration and air temperature are of major concern when considering the possible effects of global climate change on vegetations. Although production has been found to increase in many cases, other experiments have also indicated increased hazards for plant growth because of the increased frequency of weather extremes, such as droughts, floods, and extreme temperatures. Thus at the same time elevated CO(2) and the extreme climatic events, intra- and interannual climatic variability alone can be foreseen as an indirect constraint, which separately influences significantly the carbon cycling in ecosystems, too. In the shorter term the effect of CO(2) is direct and is mediated by photosynthesis. In the longer term the effects of elevated CO(2) became more and more indirect and its effects are mediated by source-sink interactions within plants, resources (nutrients, water), temperature, microbes, herbivores, and land-use management practice. In fact, the plants can make use of their general stress coping mechanisms to avoid or compensate possible negative effects of elevated CO(2). One has to consider that all the classical abiotic, biotic, and anthropogenic stressors are threatening plant growth and development also under elevated CO(2), although at possibly different doses compared to ambient CO(2) concentrations. Therefore, the knowledge of the general stress coping, stress avoiding, and tolerance mechanisms is needed to understand the regulation of the plants' metabolism under normal and elevated CO(2) levels.

Comparison of two metal surveys by moss Tortula ruralis in Budapest, Hungary.

Moss transplants of Tortula ruralis were used as active biomonitoring organisms as part of a monitoring study to assess the metals (Al, Cd, Cu, Cr, Fe, Ni, Pb, V and Zn) associated with ambient particles on mosses in Budapest, the capital town of Hungary. The moss samples were collected in a nature conservation area from a semi-arid sand grassland (Festucetum vaginatae danubiale), less than 1 month before transplantation. Moss cushions were exposed to pollution in Budapest during October-November 1993 and February-March 1994. In the study area, 16 sites were marked out as measuring sites, in accordance with the structure of the city. A similar stand was established in the Botanical Gardens of the Szent István University in Gödöllo as a control site. ICP-AES analysis of moss for metals showed the ability of Tortula ruralis to accumulate the metals under study. Control site showed lower impact in comparison to the other sites.

Photosynthetic activity of homoiochlorophyllous desiccation tolerant plant Haberlea rhodopensis during dehydration and rehydration.

The functional state of the photosynthetic apparatus of flowering homoiochlorophyllous desiccation tolerant plant Haberlea rhodopensis during dehydration and subsequent rehydration was investigated in order to characterize some of the mechanisms by which resurrection plants survive drought stress. The changes in the CO2 assimilation rate, chlorophyll fluorescence parameters, thermoluminescence, fluorescence imaging and electrophoretic characteristics of the chloroplast proteins were measured in control, moderately dehydrated (50% water content), desiccated (5% water content) and rehydrated plants. During the first phase of desiccation the net CO2 assimilation decline was influenced by stomatal closure. Further lowering of net CO2 assimilation was caused by both the decrease in stomatal conductance and in the photochemical activity of photosystem II. Severe dehydration caused inhibition of quantum yield of PSII electron transport, disappearance of thermoluminescence B band and mainly charge recombination related to S2QA- takes place. The blue and green fluorescence emission in desiccated leaves strongly increased. It could be suggested that unchanged chlorophyll content and amounts of chlorophyll-proteins, reversible modifications in PSII electron transport and enhanced probability for non-radiative energy dissipation as well as increased polyphenolic synthesis during desiccation of Haberlea contribute to drought resistance and fast recovery after rehydration.

Comparative study on the changes in photosynthetic activity of the homoiochlorophyllous desiccation-tolerant Haberlea rhodopensis and desiccation-sensitive spinach leaves during desiccation and rehydration.

The functional peculiarities and responses of the photosynthetic system in the flowering homoiochlorophyllous desiccation-tolerant (HDT) Haberlea rhodopensis and the non-desiccation-tolerant spinach were compared during desiccation and rehydration. Increasing rate of water loss clearly modifies the kinetic parameters of fluorescence induction, thermoluminescence emission, far-red induced P700 oxidation and oxygen evolution in the leaves of both species. The values of these parameters returned nearly to the control level after 24 h rehydration only of the leaves of HDT plant. PS II was converted in a non-functional state in desiccated spinach in accordance with the changes in membrane permeability, malondialdehyde, proline and H(2)O(2) contents. Moreover, our data showed a strong reduction of the total number of PS II centers in Haberlea without any changes in the energetics of the charge recombination. We consider this observation, together with the previously reported unusually high temperature of B-band (S(2)Q(B)-) emission of Haberlea to reflect some specific adaptive characteristics of the photosynthetic system. As far as we know this is the first time when such adaptive characteristics and mechanism of the photosynthetic system of a flowering HDT higher plant is described. These features of Haberlea can explain the fast recovery of its photosynthesis after desiccation, which enable this HDT plant to rapidly take advantage of frequent changes in water availability.

Steroid epimers studied by different mass spectrometric methods.

The combined use of different mass spectrometric ionization methods and MS/MS techniques provide the possibility to differentiate between stereoisomers or epimers. In this paper the mass spectral decomposition of 11alpha- and 11beta-substituted estrans was studied. Distinctive stereochemical effects have been observed in their fast atom bombardment product ion spectra. In the electron ionisation (EI) mode, the 5alpha- and 5beta-hydroxylated compounds showed significant differences in the abundance of water elimination. Mass spectrometry has proved to be an effective tool when stereoisomer steroids are compared.

Hydrolytic behavior of 5alpha-hydroxy-11beta- and 5beta-hydroxy-11alpha-substituted 19-norsteroids.

Teutsch G. and Bélanger A. treated 5alpha,10alpha epoxides with Grignard-reagents catalyzed by copper(I) ions. The reaction with steroidal epoxides proceeded with complete regio- and stereospecificity, leading exclusively to the 11beta-substituted compounds. According to our synthetic strategy, the 5,10 epoxide isomers were not separated; instead, the pure 11beta, and in some cases, 11alpha-substituted molecules were isolated after the conjugate addition of the Grignard-reagents, followed by deketalization and dehydration. Surprisingly, appearance of a third compound was generally observed beside the expected deprotected products, and this compound turned out to have a 3-keto-5(10),9(11) structural unit. Starting from pure 3-ethylenedioxy-5alpha,10alpha-epoxy-estr-9(11)-ene-17-one and 3-ethylenedioxy-5beta,10beta-epoxy-estr-9(11)-ene-17-one, four model compounds were synthesized (11alpha- and 11beta-[4-[1,1-(ethylenedioxy)-ethyl]phenyl]-estra-, as well as 11alpha- and 11beta-cyclohexyl-estra-derivatives) to study the process of deprotection and dehydration. 3-keto-5(10),9(11)-derivatives were found to form after deketalization and dehydration only from 11alpha-substituted derivatives, while 11beta-derivatives resulted in only the expected 3-keto-5,9-diene structure. After observing this remarkable difference between the behavior of 11alpha-, 11beta-substituted isomers we decided to take a closer look at the processes of deketalization and dehydration. In order to carry out the hydrolysis under mild conditions, pyridinium paratoluenesulfonate, a weakly acidic salt, was applied. All the intermediate products observed by TLC were isolated. The outcome of the deprotection and elimination reactions can be rationalized by two factors: conjugation of olefins (with the 3-oxo-group or the 11-phenyl group) and orientation of groups to be eliminated.

Seasonal changes in the levels of compatible osmolytes in three halophytic species of inland saline vegetation in Hungary.

Seasonal changes in the leaf concentration of compatible osmolytes were investigated in three halophytic species (Lepidium crassifolium, Camphorosma annua and Limonium gmelini subsp. hungaricum) native to a salty-sodic grassland. The investigated species were shown to accumulate both carbohydrate- and amino acid-derived osmolytes. The leaf tissues of C. annua (Chenopodiaceae) preferentially stored glycine betaine and pinitol, while in L. gmelini (Plumbaginaceae) beta-alanine betaine, choline-O-sulphate, and pinitol were accumulated. In the leaves of L. crassifolium (Brassicaceae) a very high amount of proline, associated with a high level of soluble carbohydrates was found. Not only the biochemical nature of the osmolyte, but also the seasonal pattern of osmolyte accumulation showed significant species-specific fluctuations. In addition, the cellular levels of the observed osmolytes changed with the growth period and according to the environmental parameters. The highest concentrations of osmolytes were found in March, when low temperatures, hypoxic conditions and high salt concentrations were the main constraints to plant growth. The high structural diversity of osmolytes combined with their multifunctionality and the seasonal flexibility of the metabolism in plants facing multiple stresses is discussed.

Investigations on ecological effects of heavy metal pollution in Hungary by moss-dwelling water bears (Tardigrada), as bioindicators.

The authors demonstrate a possible relationship between the concentrations of metals (Cd, Cr, Cu, Fe, Ni, Pb, V, Zn) measured in mosses and the composition of Tardigrade species detected in the same samples. Cushions of Hypnum cupressiforme were collected at 18 sites distributed in the whole of Hungary to estimate the background air pollution, then analyzed by ICP-AES to determine the concentrations of heavy metals. Data reflect the expected correlation; elevated heavy metal contents decrease the number of water bear species and of specimens, and abundance of Tardigrada depends strongly on air pollution. Higher concentrations of cadmium and chromium seem to have particularly damaging and toxic influence on community structure. As the biologically relevant effects of pollution can only be evaluated by carrying out measurements on the organisms themselves, our method applied species of Bryophyte and of Tardigrade, as bioindicators were appeared to be an adequate method to show the effect of air pollution on abundance of water bears.

Synthesis and structure-activity relationships of neuromuscular blocking agents.

The first use of neuromuscular blocking agents (muscle relaxants) in clinical practice (1942) revolutionised the practice of anaesthesia and started the modern era of surgery. Since 1942 introduction of tubocurarine (18) neuromuscular blocking agents have been used routinely to provide skeletal muscle relaxation during surgical procedures allowing access to body cavities without hindrance from voluntary or reflex muscle movement. After the introduction of tubocurarine and the depolarizing suxamethonium chloride (4) (1949) several nondepolarizing steroidal and nonsteroidal neuromuscular blocking agents with different onset time and duration of effect were introduced e.g. gallamine triethiodide (1) (1949), methocurine (2) (1949), alcuronium chloride (3) (1963), pancuronium bromide (9) (1968), vecuronium bromide (11) (1982), pipecuronium bromide (10) (1982), atracurium besylate (5) (1982), doxacurium chloride (6) (1991), mivacurium chloride (8) (1992), rocuronium bromide (12) (1994) cisatracurium besylate (7) (1996), and rapacuronium bromide (13) (2000). SZ 1677 (14) a steroid type nondepolarizing neuromuscular blocking agent under development (preclinical phase). This review article deals with a comprehensive survey of the progress in chemical, pharmacological and, in some respects, of clinical studies of neuromuscular blocking agents used in the clinical practice and under development, including the synthesis, structure elucidation, pharmacological actions, structure activity relationships studies of steroidal and nonsteroidal derivatives.

Microwave-assisted Stille-coupling of steroidal substrates.

Steroidal dienes were synthesised by Stille-coupling of the corresponding alkenyl iodides with vinyltributyltin under microwave irradiation in a domestic microwave oven in drastically reduced reaction times. Rate acceleration was observed also in the one-pot Stille-coupling-Diels-Alder reaction of 17-iodo-5alpha-androst-16-ene. Stereoselectivity of cycloaddition was slightly improved with diethyl maleate as the dienophile, compared to that achieved with thermal heating.

Facile synthesis of 17-formyl steroids via palladium-catalyzed homogeneous carbonylation reaction.

17-formyl-androst-16-ene and its analogues were synthesized from the corresponding 17-iodo-16-ene derivatives in palladium-catalyzed formylation reaction using tributyltin hydride as hydrogen source under mild reaction conditions. The formation of androst-16-ene and its isomerization products, as well as that of analogous steroidal olefins as side-products, was found to be dependent on the reaction conditions. The formylation reaction tolerates various functional groups on the A and B rings of the steroids.

Synthesis of steroidal diacyl hydrazines and their 1,3,4-oxadiazole derivatives.

Homogeneous catalytic hydrazinocarbonylation of some steroid derivatives possessing iodo-alkenyl moiety (17-iodo-androst-16-ene 1, 17-iodo-3-methoxy-estra-1,3,5(10),16-tetraene 2, 17-iodo-4-aza-4-methyl-androst-16-en-3-one 3 and 17-iodo-6beta-hydroxy-3alpha,5alpha-cycloandrost-16-ene 4) were carried out in the presence of a palladium catalyst, a base and acetic or benzoic hydrazide as the nucleophilic reagent. The corresponding N-acetamido-carbamoyl 1a-4a or N-benzamido-carbamoyl derivatives 1b-4b were obtained in high yields. Some of these derivatives served as starting materials for the synthesis of new steroidal 1,3,4-oxadiazole compounds.

Ecophysiological consequences of contrasting microenvironments on the desiccation tolerant moss Tortula ruralis.

Tortula ruralis is a homoiochlorophyllous-desiccation-tolerant (HDT) moss that retains all pigments when dehydrated and rapidly recovers physiological function upon rehydration. This moss forms extensive cover in exposed and shaded areas in the sandy semi-arid grasslands of Central Europe. We hypothesized that contrasting drying regimes between these microhabitats would affect plant N status, constraints to gas exchange and growth, as well as result in altered pigment concentrations and ratios, and photochemical light-response dynamics. Furthermore, we believed T. ruralis's HDT habit would limit its ability to acclimate to altered light environment. We found that sun plant T. ruralis had lower plant mass, as well as lower tissue N, C, total photosynthetic pigment concentrations and carbon isotope discrimination (Δ) values compared to shade plant counterparts. Carotenoid/chlorophyll ratios in sun plants were typical of high light-adapted tissue, but chlorophyll a/chlorophyll b ratios were lower, more characteristic of low light-adapted tissue. This unique combination of pigment responses was accompanied by sustained lower levels of optimal quantum efficiency of PSII (F v/F m) in sun plant T. ruralis, even during favorable diurnal conditions, and reduced engagement of energy-dependent thermal dissipation (NPQ). Reciprocal transplants of sun and shade plants showed that T. ruralis is capable of short-term adjustment to altered light level, as evidenced by increases in F v/F m, NPQ, and light-adapted PSII yield (φPSII) in transplanted sun plants, and concurrent decreases in sun-transplanted shade plants. However, the performance of transplanted sun plants remained consistently below that of undisturbed shade plants. These findings show that microenvironmental variation results in different patterns of resource acquisition in this HDT moss, and that growth in the open imparts greater desiccation tolerance, and the development of a greater standing engagement of slowly reversing photoprotective mechanisms. In contrast, prolonged activity and greater resource acquisition in shaded populations may allow T. ruralis to rapidly adjust to changes following disturbance to the plant canopy, fostering the persistence of T. ruralis in these semi-arid grasslands.

Poikilohydry and homoihydry: antithesis or spectrum of possibilities?

Plants have followed two principal (and contrasting) strategies of adaptation to the irregular supply of water on land, which are closely bound up with scale. Vascular plants evolved internal transport from the soil to the leafy canopy (but their 'homoihydry' is far from absolute, and some are desiccation tolerant (DT)). Bryophytes depended on desiccation tolerance, suspending metabolism when water was not available; their cells are generally either fully turgid or desiccated. Desiccation tolerance requires preservation intact through drying-re-wetting cycles of essential cell components and their functional relationships, and controlled cessation and restarting of metabolism. In many bryophytes and some vascular plants tolerance is essentially constitutive. In other vascular plants (particularly poikilochlorophyllous species) and some bryophytes tolerance is induced by water stress. Desiccation tolerance is adaptively optimal on hard substrates impenetrable to roots, and on poor dry soils in seasonally dry climates. DT vascular plants are commonest in warm semiarid climates; DT mosses and lichens occur from tropical to polar regions. DT plants vary widely in their inertia to changing water content. Some mosses and lichens dry out and recover within an hour or less; vascular species typically respond on a time scale of one to a few days. Contents Summary 327 I. Introduction 328 II. The soil-plant-atmosphere continuum 329 III. Desiccation-tolerant plants: taxonomic distribution and functional characteristics 331 IV. Anatomical and physiological requirements and implications of desiccation tolerance 333 V. Time-scale considerations and ecological adaptation 340 VI. Conclusion 344 Acknowledgements 344 References 344.

Photosynthetic responses of a moss, Tortula ruralis, ssp. ruralis, and the lichens Cladonia convoluta and C. furcata to water deficit and short periods of desiccation, and their ecophysiological significance: a baseline study at present-day CO2 concentration.

We report the changes in CO2 assimilation, potential photochemical activity (as measured by slow fluorescence), photosynthetic pigment concentrations, and dark respiration of two desiccation-tolerant (DT) lichens (Cladonia convoluta (Lam.) P. Cout. and C. furcata (Huds.) Schrad.), and a DT moss (Tortula ruralis (Hedw.) Gaertn. ssp. ruralis) during slow drying, and on rehydration following a 12 h period of desiccation. Initially there was a two to fourfold increase in net CO., assimilation due to reduction of CO2 -diffusion resistance by elimination of excess water. Optimum water content for photosynthesis was 100-150 % of dry mass (DM) in C. convoluta, c. 100 % DM in C. furcata, and 120-200 % DM in T. ruralis. The intensity of maximum and steady-state slow fluorescence showed little change above water contents of 56%, DM in the lichens and 73 % DM in T. ruralis (corresponding to c. 30-40 % cell relative water content), but fell sharply at lower water content. The variable duorophyll-fluorescence decrease ratio (Rfd) at 690 nm peaked at 56 % DM water content in the two lichens, and at 45% DM in T. ruralis. Photochemical activity ceased at the same point in the experiments as CO, assimilation; dark respiration ceased only when desiccation was complete. In all three species, the photosynthetic apparatus remained in a fully and quickly recoverable state. Chlorophyll and carotenoid concentrations remained substantially unaltered throughout. On rehydration, chlorophyll fluorescence parameters returned within 30 min to pre-desiccation levels, and photosynthesis recovered fully and rapidly (< 1 h). All three species attained a positive carbon balance within 20 min of re-moistening, in spite of high rates of dark respiration. The results confirm the significance of extracellularly-stored water to poikilohydric DT lichens and bryophytes. The measurements, in conjunction with published data on the full-turgor water content of similar mosses and lichens, show that the cell-physiological response of photosynthesis to water deficit is not greatly different from that of either normal or DT vascular plants. Small plant size and small cell volume in DT lichens and mosses, together with rapid recovery of photosynthesis after desiccation, allow the plants to utilize the small amounts of intermittently available water from brief showers or dew.