Lipids are fundamental components of cell membranes and play a significant role in their integrity and fluidity. Alteration in lipid composition of membranes has been reported to be a major response ...to abiotic environmental stresses. This work was focused on the characterization of frond lipid composition and membrane integrity during a desiccation–rehydration cycle of two filmy fern species with contrasting desiccation tolerance: Hymenophyllum caudiculatum (less tolerant) and Hymenophyllum plicatum (more tolerant). The relative water content decreased without differences between species when both filmy ferns were subjected to desiccation. However, H. plicatum reached a higher relative water content than H. caudiculatum after rehydration. Fatty acids profiles showed the presence of a very long chain polyunsaturated fatty acid during the desiccation–rehydration cycle, with eicosatrienoic acid being the most abundant. Additionally, propidium iodide permeation staining and confocal microscopy demonstrated that, following the desiccation–rehydration cycle, H. plicatum exhibited a greater membrane integrity than H. caudiculatum. The lack of some very long chain fatty acids such as C22:1n9 and C24:1n9 in this species contrasting with H. plicatum may be associated with its lower membrane stability during the desiccation–rehydration cycle. This report provides the first insight into the fatty acid composition and dynamics of the membrane integrity of filmy ferns during a desiccation–rehydration cycle. This could potentially play a role in determining the different levels of desiccation tolerance and microhabitat preferences exhibited by Hymenophyllaceae species.
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 ${\mathrm{S}}_{2}{\mathrm{A}}_{\mathrm{A}}^{-}$ 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.
The changes in some proteins involved in the light reactions of photosynthesis of the resurrection plant
Haberlea rhodopensis were examined in connection with desiccation. Fully hydrated (control) ...and completely desiccated plants (relative water content (RWC) 6.5%) were used for thylakoid preparations. The chlorophyll (Chl) a to Chl b ratios of thylakoids isolated from control and desiccated leaves were very similar, which was also confirmed by measuring their absorption spectra. HPLC analysis revealed that β-carotene content was only slightly enhanced in desiccated leaves compared with the control, but the zeaxanthin level was strongly increased. Desiccation of
H. rhodopensis to an air-dried state at very low light irradiance led to a little decrease in the level of D1, D2, PsbS and PsaA/B proteins in thylakoids, but a relative increase in LHC polypeptides. To further elucidate whether the composition of the protein complexes of the thylakoid membranes had changed, we performed a separation of solubilized thylakoids on sucrose density gradients. In contrast to spinach,
Haberlea thylakoids appeared to be much more resistant to the same solubilization procedure, i.e. complexes were not separated completely and complexes of higher density were found. However, the fractions analyzed provided clear evidence for a move of part of the antenna complexes from PSII to PSI when plants became desiccated. This move was also confirmed by low temperature emission spectra of thylakoids.
Overall, the photosynthetic proteins remained comparatively stable in dried
Haberlea leaves when plants were desiccated under conditions similar to their natural habitat. Low light during desiccation was enough to induce a rise in the xanthophyll zeaxanthin and β-carotene. Together with the extensive leaf shrinkage and some leaf folding, increased zeaxanthin content and the observed shift in antenna proteins from PSII to PSI during desiccation of
Haberlea contributed to the integrity of the photosynthetic apparatus, which is important for rapid recovery after rehydration.
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.
Haberlea rhodopensis plants, growing under low irradiance in their natural habitat, were desiccated to air-dry state at a similar light intensity (about 30 μmol m⁻² s⁻¹) under optimal (23/20°C, ...day/night) or high (38/30°C) temperature. Dehydration of plants at high temperature increased the rate of water loss threefold and had a more detrimental effect than either drought or high temperature alone. Water deficit decreased the photochemical activity of PSII and PSI and the rate of photosynthetic oxygen evolution, and these effects were stronger when desiccation was carried out at 38°C. Some reduction in the amount of the main PSI and PSII proteins was observed especially in severely desiccated Haberlea leaves. The results clearly showed that desiccation of the homoiochlorophyllous poikilohydric plant Haberlea rhodopensis at high temperature had more damaging effects than desiccation at optimal temperature and in addition recovery was slower. Increased thermal energy dissipation together with higher proline and carotenoid content in the course of desiccation at 38°C compared to desiccation at 23°C probably helped in overcoming the stress.
The effect of high irradiance (HI) during desiccation and subsequent rehydration of the homoiochlorophyllous desiccation-tolerant shade plant Haberlea rhodopensis was investigated. Plants were ...irradiated with a high quantum fluence rate (HI; 350 μmol m⁻² s⁻¹ compared to ca. 30 μmol m⁻² s⁻¹ at the natural rock habitat below trees) and subjected either to fast desiccation (tufts dehydrated with naturally occurring thin soil layers) or slow desiccation (tufts planted in pots in peat-soil dehydrated by withholding irrigation). Leaf water content was 5 % of the control after 4 d of fast and 19 d of slow desiccation. Haberlea was very sensitive to HI under all conditions. After 19 d at HI, even in well-watered plants there was a strong reduction of rates of net photosynthesis and transpiration, contents of chlorophyll (Chl) and carotenoids, as well as photosystem 2 activity (detected by the Chl fluorescence ratio RFd). Simultaneously, the blue/red and green/red fluorescence ratios increased considerably suggesting increased synthesis of polyphenolic compounds. Desiccation of plants in HI induced irreversible changes in the photosynthetic apparatus and leaves did not recover after rehydration regardless of fast or slow desiccation. Only young leaves survived desiccation.
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.
Haberlea rhodopensis (Gesneriaceae) is endemic plant in a very small group of poikilohydric angiosperms that are able to survive in an almost complete dehydrate state. Upon watering the plants ...rapidly revive and are restored to their former state. H. rhodopensis belongs to the homoiochlorophyllous type of desiccation tolerant plants that keep their chlorophyll content during drying. The high amount of chlorophyll molecules retained during desiccation could be a source for potentially harmful singlet oxygen production. That is why it was important to investigate the changes in the activity of some antioxidant enzymes during dehydration of Haberlea. Plants were desiccated at optimal (23 °C) and high (38 °C) temperature as drought stress is frequently accompanied by high temperatures under field conditions. The results showed that superoxide dismutase (SOD; EC 1.15.1.1) activity gradually decreased, whereas catalase (EC 1. 11. 1. 6) activity significantly increased during desiccation of Haberlea plants under both optimal and high temperature. Exposure of plants to high temperature reduced the activity of these enzymes. The enhanced activity of ascorbate peroxidase (APX; EC 1.11.1.11) and guaiacol peroxidase (GPX; EC 1.11.1.7) was observed under moderate water stress, after which they declined. High temperature stress applied alone did not influence the APX and GPX activity.
Inhibition of photosynthesis is a central, primary response that is observed in both desiccation-tolerant and desiccation-sensitive plants affected by drought stress. Decreased photosynthesis during ...drought stress can either be due to the limitation of carbon dioxide entry through the stomata and the mesophyll cells, due to increased oxidative stress or due to decreased activity of photosynthetic enzymes. Although the photosynthetic rates decrease in both desiccation-tolerant and sensitive plants during drought, the remarkable difference lies in the complete recovery of photosynthesis after rehydration in desiccation-tolerant plants. Desiccation of sensitive plants leads to irreparable damages of the photosynthetic membranes, in contrast the photosynthetic apparatus is deactivated during desiccation in desiccation-tolerant plants. Desiccation-tolerant plants employ different strategies to protect and/or maintain the structural integrity of the photosynthetic apparatus to reactivate photosynthesis upon water availability. Two major mechanisms are distinguished. Homoiochlorophyllous desiccation-tolerant plants preserve chlorophyll and thylakoid membranes and require active protection mechanisms, while poikilochlorophyllous plants degrade chlorophyll in a regulated manner but then require de novo synthesis during rehydration. Desiccation-tolerant plants, particularly homoiochlorophyllous plants, employ conserved and novel antioxidant enzymes/metabolites to minimize the oxidative damage and to protect the photosynthetic machinery. De novo synthesized, stress-induced proteins in combination with antioxidants are localized in chloroplasts and are important components of the protective network. Genome sequence informations provide some clues on selection of genes involved in protecting photosynthetic structures; e.g. ELIP genes (early light inducible proteins) are enriched in the genomes and more abundantly expressed in homoiochlorophyllous desiccation-tolerant plants. This review focuses on the mechanisms that operate in the desiccation-tolerant plants to protect the photosynthetic apparatus during desiccation.
Tolerance to near complete desiccation of vegetative organs is a widespread capability in bryophytes and is also shared by a small group of vascular plants known as resurrection plants. To date more ...than 300 species, belonging to pteridophytes and angiosperms, have been identified that possess this kind of desiccation-tolerance. The vegetative desiccation-tolerance of resurrection plants is an inductive process displayed only under environmental stress with or without the involvement of abscisic acid as molecular signal. The different problems associated with desiccation encountered by resurrection plants render the employment of many interacting mechanisms necessary. Preservation of cell order and correct structure of membranes and macromolecules is underpinned by the synthesis of large amounts of sugars, amino acids, and small polypeptides such as late embryogenesis abundant (LEA) proteins and dehydrins. Some of these compatible solutes, such as sucrose and LEA proteins, are also involved in cytoplasm vitrification, which occurs during the last phase of desiccation. Mechanical damage due to vacuole shrinkage in dehydrating cells is avoided by cell wall folding or by replacing the water in vacuoles with nonaqueous substances. Oxidative stress, due to enhanced production of reactive oxygen species (ROS) especially by chloroplasts, is minimized through two different strategies. The homoiochlorophyllous resurrection plants, which conserve chloroplasts with chlorophylls and thylakoids upon drying, fold leaf blades and synthesize anthocyanins, as both sunscreens and free radical scavengers, and additionally increase the activity of antioxidant systems in cells. In contrast, the chloroplasts in poikilochlorophyllous species degrade chlorophylls and thylakoid membranes yielding desiccoplasts that are devoid of any internal structures. These adaptive mechanisms preserve cells from damage by desiccation and allow them to resume vital functions once rehydrated. Even if based mainly on cell protection during drying, the vegetative desiccation-tolerance of resurrection plants also relies on systems of cell recovery and repair upon rehydration. However, most of these systems are prepared during cell dehydration.
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