Differential proteomics of dehydration and rehydration in bryophytes: evidence towards a common desiccation tolerance mechanism

All bryophytes evolved desiccation tolerance (DT) mechanisms during the invasion of terrestrial habitats by early land plants. Are these DT mechanisms still present in bryophytes that colonize aquatic habitats? The aquatic bryophyte Fontinalis antipyretica Hedw. was subjected to two drying regimes and alterations in protein profiles and sucrose accumulation during dehydration and rehydration were investigated. Results show that during fast dehydration, there is very little variation in protein profiles, and upon rehydration proteins are leaked. On the other hand, slow dehydration induces changes in both dehydration and rehydration protein profiles, being similar to the protein profiles displayed by the terrestrial bryophytes Physcomitrella patens (Hedw.) Bruch and Schimp. and, to what is comparable with Syntrichia ruralis (Hedw.) F. Weber and D. Mohr. During dehydration there was a reduction in proteins associated with photosynthesis and the cytoskeleton, and an associated accumulation of proteins involved in sugar metabolism and plant defence mechanisms. Upon rehydration, protein accumulation patterns return to control values for both photosynthesis and cytoskeleton whereas proteins associated with sugar metabolism and defence proteins remain high. The current results suggest that bryophytes from different ecological adaptations may share common DT mechanisms.     

Hardening by partial dehydration and ABA increase desiccation tolerance in the cyanobacterial lichen Peltigera polydactylon

* BACKGROUND AND AIMS: The ability of partial dehydration and abscisic acid pretreatments to increase desiccation tolerance in the cyanobacterial lichen Peltigera polydactylon was tested. * METHODS: Net photosynthesis and respiration were measured using infrared gas analysis during a drying and rehydration cycle. At the same time, the efficiency of photosystem two was measured using chlorophyll fluorescence, and the concentrations of chlorophyll a were spectrophotometrically assayed. Heat production was also measured during a shorter drying and rehydration cycle using differential dark microcalorimetry. * KEY RESULTS: Pretreating lichens by dehydrating them to a relative water content of approx. 0.65 for 3 d, followed by storing thalli hydrated for 1 d in the light, significantly improved their ability to recover net photosynthesis during rehydration after desiccation for 15 but not 30 d. Abscisic acid pretreatment could substitute for partial dehydration. The improved rates of photosynthesis during the rehydration of pretreated material were not accompanied by preservation of photosystem two activity or chlorophyll a concentrations compared with untreated lichens. Partial dehydration and ABA pretreatments appeared to have little direct effect on the desiccation tolerance of the mycobiont, because the bursts of respiration and heat production that occurred during rehydration were similar in control and pretreated lichens. * CONCLUSIONS: Results indicate that the photobiont of P. polydactylon possesses inducible tolerance mechanisms that reduce desiccation-induced damage to carbon fixation, and will therefore improve the supply of carbohydrates to the whole thallus following stress. In this lichen, ABA is involved in signal transduction pathways that increase tolerance of the photobiont.     

Increased activities of superoxide dismutase and catalase are not the mechanism of desiccation tolerance induced by hardening in the moss Atrichum androgynum

Abstract

The effects of treatments that increase desiccation tolerance were tested on the activity of the enzymes superoxide dismutase (SOD) and catalase (CAT) in the moss Atrichum androgynum subjected to a drying/wetting cycle. Hardening by both abscisic acid (ABA) pretreatment and partial dehydration significantly increased the rate of recovery of photosynthesis during rehydration following desiccation. Hardening treatments had little effect on SOD activity. In non-hardened plants, SOD activity increased three-fold during desiccation for 32 h at 52% rh, but hardened material tended to display smaller increases in activity. During rehydration, SOD activities rapidly declined to their initial values in all treatments. Hardening by partial dehydration, but not ABA, reduced CAT activity. After desiccation for 32 h, material from all treatments displayed about half the initial CAT activity, and activity did not change during subsequent rehydration. Results show that, while the induction of SOD appears to play a role in desiccation tolerance, a similar induction occurred in both hardened and non-hardened mosses. Induction of greater activities of enzymes that scavenge reactive oxygen species is not responsible for the added tolerance induced by hardening treatments.     

Physiological consequences of desiccation in the aquatic bryophyte <Emphasis Type="Italic">Fontinalis antipyretica</Emphasis>

The moss Fontinalis antipyretica, an aquatic bryophyte previously described as desiccation-intolerant, is known to survive intermittent desiccation events in Mediterranean rivers. To better understand the mechanisms of desiccation tolerance in this species and to reconcile the apparently conflicting evidence between desiccation tolerance classifications and field observations, gross photosynthesis and chlorophyll a fluorescence were measured in field-desiccated bryophyte tips and in bryophyte tips subjected in the laboratory to slow, fast, and very fast drying followed by either a short (30 min) or prolonged (5 days) recovery. Our results show, for the first time, that the metabolic response of F. antipyretica to desiccation, both under field and laboratory conditions, is consistent with a desiccation-tolerance pattern; however, drying must proceed slowly for the bryophyte to regain its pre-desiccation state following rehydration. In addition, the extent of dehydration was found to influence metabolism whereas the drying rate determined the degree of recovery. Photosystem II (PSII) regulation and structural maintenance may be part of the induced desiccation tolerance mechanism allowing this moss to recover from slow drying. The decrease in the photochemical quenching coefficient (qP) immediately following rehydration may serve to alleviate the effects of excess energy on photosystem I (PSI), while low-level non-photochemical quenching (NPQ) would allow an energy shift enabling recovery subsequent to extended periods of desiccation. The findings were confirmed in field-desiccated samples, whose behavior was similar to that of samples slowly dried in the laboratory.     

Effects of Dehydration and Rehydration on Photosynthesis of Detached Leaves of the Resurrective Plant Boea hygrometrica

Fluorescence induction kinetics was used to investigate the effects of dehydration and rehydration on photosynthesis of detached leaves of the desiccation-tolerant, resurrective plant Boea hygrometrica (Bunge) R. Br. In comparison with the desiccation-intolerant plant Chirita heterotricha Merr., the PSⅡphotochemical activity of Boea hygrometrica was characterized by a faster decline during dehydration and a much higher capacity of recovery during rehydration. By means of native PAGE, it was further shown that the thylakoid pigment-protein complexes of Boea hygrometrica were highly stable during dehydration and rehydration. These features may contribute to the extreme desiccation resistance of photosynthesis apparatus of resurrective plant Boea hygrometrica.     

Photosynthetic limitations and volatile and non‐volatile isoprenoids in the poikilochlorophyllous resurrection plant Xerophyta humilis during dehydration and rehydration

We investigated the photosynthetic limitations occurring during dehydration and rehydration of Xerophyta humilis, a poikilochlorophyllous resurrection plant, and whether volatile and non‐volatile isoprenoids might be involved in desiccation tolerance. Photosynthesis declined rapidly after dehydration below 85% relative water content (RWC). Raising intercellular CO₂ concentrations during desiccation suggest that the main photosynthetic limitation was photochemical, affecting energy‐dependent RuBP regeneration. Imaging fluorescence confirmed that both the number of photosystem II (PSII) functional reaction centres and their efficiency were impaired under progressive dehydration, and revealed the occurrence of heterogeneous photosynthesis during desiccation, being the basal leaf area more resistant to the stress. Full recovery in photosynthetic parameters occurred on rehydration, confirming that photosynthetic limitations were fully reversible and that no permanent damage occurred. During desiccation, zeaxanthin and lutein increased only when photosynthesis had ceased, implying that these isoprenoids do not directly scavenge reactive oxygen species, but rather protect photosynthetic membranes from damage and consequent denaturation. X. humilis was found to emit isoprene, a volatile isoprenoid that acts as a membrane strengthener in plants. Isoprene emission was stimulated by drought and peaked at 80% RWC. We surmise that isoprene and non‐volatile isoprenoids cooperate in reducing membrane damage in X. humilis, isoprene being effective when desiccation is moderate while non‐volatile isoprenoids operate when water deficit is more extreme.     

Reversible cellular and metabolic changes induced by dehydration in desiccation-tolerant wheat seedling shoots

Wheat seedlings obtained after 2 or 3 days of seed germination in darkness at 20°C (i.e. with a 0.5–0.7 cm long coleoptile) were still viable after drying in darkness in ambient conditions which reduced the shoot moisture content to about 0.30 g H₂O g^?1 dry mass (DM). Coleoptile and primary leaf growth resumed upon rehydration, but primary roots died and new roots regenerated. In the present work we have investigated whether desiccation tolerance of the shoot (coleoptile and primary leaf combined) was related to some reversible cellular or metabolic changes induced by dehydration. Non‐dehydrated shoots were high in moisture content (4.0–5.0 g H₂O g^?1 DM) and exhibited an active metabolism as indicated by a high energy charge (EC = 0.85) and cells with well developed mitochondria, endoplasmic reticulum, polysomes and Golgi bodies. Dehydration induced changes in cell membrane properties since it reduced in vivo capacity of the shoot to convert 1‐aminocyclopropane 1‐carboxylic acid (ACC) to ethylene (i.e. ACC oxidase activity). This effect was already observed at 4–5 h of dehydration, namely when shoot moisture content dropped down below about 3.0 g H₂O g^?1 DM, and ACC‐dependent ethylene production became almost nil when shoot moisture content reached 1.0 g H₂O g^?1 DM. Dehydration also resulted in decreases in ATP and non‐adenylic triphosphate nucleotide (NTP) contents down to 1–2% of their initial values, and in EC value to 0.20. Concomitant with water loss, sucrose content of the shoot increased and was maximal (about 330 mg g^?1 DM, namely three‐fold that of non‐dehydrated organs) after 2 days of drying. Upon rehydration, shoots regained their original moisture content within 3 days, during which they progressively recovered apparent normal metabolism. Reversal of extensive dehydration‐associated cell wall folding occurred between 2 and 3 days of rehydration, when the ultrastructure of coleoptile and primary leaf cells also provided evidence of intensive autophagic activity, indicative of the removal of damaged cell components. Concomitantly, apparently undamaged organelles and endomembranes persisted in the cytoplasm. Restoration of 60–70% of ACC oxidase activity and 80–90% of EC value occurred within 48 and 18 h, respectively. However, the values of the ATP/ADP and NTP/ATP ratios remained lower than in control non‐dehydrated shoots, indicating that not all metabolic deterioration induced by dehydration was completely repaired. Differences in relationships between shoot moisture content and ACC‐oxidase activity or energy metabolism during dehydration and upon rehydration, and cell ultrastruture analyses suggest that desiccation tolerance of wheat seedling shoot is related to mechanisms involved in the maintenance of cell structure during water loss and the cell capacity to repair the dehydration damage.     

Evidence for the presence of photorespiration in desiccation-sensitive leaves of the C4 'resurrection' plant Sporobolus stapfianus during dehydration stress

The possible role of photorespiration as a general stress protectionmechanism, and in C₄ plant metabolism, is controversial. Inparticular, the potential involvement of photorespiration inthe acquisition of desiccation tolerance in ‘resurrection’plants is unknown. An investigation was carried out into whetherphotorespiration is present in leaves of the C₄ resurrectionplant Sporobolus stapfianus Gandoger (Poaceae) and whether itfunctions as a mechanism of stress resistance in the desiccation-tolerantyounger leaves (YL) of this plant. It is shown that the enzymesinvolved in the photorespiratory pathway maintain their activityuntil 88% relative water content (RWC) in both YL and desiccation-sensitiveolder leaves (OL). In subsequent stages of dehydration stress,the enzymatic activity declined similarly in both YL and OL.The content of the phorespiratory metabolite, serine, and ethanolamine,a direct product of serine decarboxylation, is higher in theearly stages of dehydration (88% RWC) in OL, suggesting a transientlyenhanced photorespiratory activity in these leaves. This wasconfirmed by simultaneous gas exchange and fluorescence measurements,showing suppression of the electron transport rate in OL exposedto non-photorespiratory conditions (2% O₂) at 85% RWC. It isconcluded that a higher photorespiratory electron transportoccurs in desiccation-sensitive OL, and it is therefore proposedthat the capacity to scavenge excess electrons through photorespirationdoes not contribute to protect leaves of the desiccation-tolerantYL of S. stapfianus during the stress. Key words: Ethanolamine, glycine, photorespiratory enzymes, photosynthesis, poikilohydric plant, serine Received 5 June 2007; Revised 3 September 2007 Accepted 17 September 2007     

Proteome analysis of leaves from the resurrection plant <Emphasis Type="Italic">Boea hygrometrica</Emphasis> in response to dehydration and rehydration

Resurrection plants differ from other species in their unique ability to survive desiccation. In order to understand the mechanisms of desiccation tolerance, proteome studies were carried out using leaves of the resurrection plant Boea hygrometrica to reveal proteins that were differentially expressed in response to changes in relative water content. This opportunity was afforded by the rare ability of excised B. hygrometrica leaves to survive and resume metabolism following desiccation in a manner similar to intact plants. From a total of 223 proteins that were reproducibly detected and analyzed, 35% showed increased abundance in dehydrated leaves, 5% were induced in rehydrated leaves and 60% showed decreased or unchanged abundance in dehydrated and rehydrated leaves. Since the induction kinetics fall into clearly defined patterns, we suggest that programmed regulation of protein expression triggered by changes of water status. Fourteen dehydration responsive proteins were analyzed by mass spectrometry. Eight proteins were classified as playing a role in reactive oxygen species scavenging, photosynthesis and energy metabolism. In agreement with these findings, glutathione content and polyphenol oxidase activity were found to increase upon dehydration and rapid recovery of photosynthesis was observed.     

Amino acid pattern and glutamate metabolism during dehydration stress in the 'resurrection' plant Sporobolus stapfianus: a comparison between desiccation-sensitive and desiccation-tolerant leaves

The present study analyses changes in nitrogen compounds, amino acid composition, and glutamate metabolism in the resurrection plant Sporobolus stapfianus during dehydration stress. Results showed that older leaves (OL) were desiccation-sensitive whereas younger leaves (YL) were desiccation-tolerant. OL lost their soluble protein more rapidly, and to a larger extent than YL. Enzymes of primary nitrogen assimilation were affected by desiccation and the decrease in the glutamine synthetase (GS, EC 6.3.1.2) and ferredoxin-dependent GOGAT (Fd-GOGAT, EC 1.4.7.1) activities was higher in OL than in YL, thus suggesting higher sensibility to dehydration. Moreover, YL showed higher total GS enzyme activity at the end of the dehydration stress and was shown to maintain high chloroplastic GS protein content during the entire stress period. Free amino acid content increased in both YL and OL between 88% and 6% relative water content. Interestingly, OL and YL did not accumulate the same amino acids. OL accumulated large amounts of proline and gamma-aminobutyrate whereas YL preferentially accumulated asparagine and arginine. It is concluded (i) that modifications in the nitrogen and amino acid metabolism during dehydration stress were different depending on leaf development and (ii) that proline and gamma-aminobutyrate accumulation in S. stapfianus leaves were not essential for the acquisition of desiccation tolerance. On the contrary, the accumulation of large amounts of asparagine and arginine in the YL during dehydration could be important and serve as essential nitrogen and carbon reservoirs useful during rehydration. In this context, the role of GS for asparagine accumulation in YL is discussed.     

更苏被子植物的光合作用

更苏植物是一类在极度干燥条件下组织会迅速脱水后遇水又能很快复苏的植物.极少数被子植物有这种能力,在双子叶植物中尤其罕见,而且脱水时叶绿素含量和叶绿体完整性变化较少,称为叶绿素保持型(HDT).该类植物的复苏机理简单,研究方便,因而得到更广泛注意.更苏被子植物光合作用的最新研究进展说明,光化学活性是研究更苏植物脱水复苏生理状态的灵敏指标.和普通植物一样,在光下,更苏被子植物的光化学活性随着叶片失水而受到抑制,但奇怪的是在失去95%以上的水分后复水仍可迅速复活.在脱水过程中叶黄素循环和抗氧化系统的上调以及光合膜完整性和稳定性的保持,可能对更苏被子植物的耐脱水性起非常重要的作用.磷酸盐对复苏的影响也表现在复水阶段而且与上述两种保护机理关系不大,因此应该加强更苏被子植物复水阶段的研究.     

Dynamics of Endogenous Phytohormones during Desiccation and Recovery of the Resurrection Plant Species Haberlea rhodopensis

Drought is one of the most significant threats to world agriculture and hampers the supply of food and energy. The mechanisms of drought responses can be studied using resurrection plants that are able to survive extreme dehydration. As plant hormones function in an intensive cross-talk, playing important regulatory roles in the perception and response to unfavorable environments, the dynamics of phytohormones was followed in the resurrection plant Haberlea rhodopensis Friv. during desiccation and subsequent recovery. Analysis of both leaves and roots revealed that jasmonic acid, along with and even earlier than abscisic acid, serves as a signal triggering the response of the resurrection plants to desiccation. The steady high levels of salicylic acid could be considered an integral part of the specific set of parameters that prime H. rhodopensis desiccation tolerance. The dynamic changes of cytokinins and auxins suggest that these hormones actively participate in the dehydration response and development of desiccation tolerance in the resurrection plants. Our data contribute to the elucidation of a global complex picture of the resurrection plant’s ability to withstand desiccation, which might be successfully utilized in crop improvement.     

更苏被子植物的光合作用

更苏植物是一类在极度干燥条件下组织会迅速脱水后遇水又能很快复苏的植物。极少数被子植物有这种能力,在双子叶植物中尤其罕见,而且脱水时叶绿素含量和叶绿体完整性变化较少,称为叶绿素保持型(HDT)。该类植物的复苏机理简单,研究方便,因而得到更广泛注意。更苏被子植物光合作用的最新研究进展说明,光化学活性是研究更苏植物脱水复苏生理状态的灵敏指标。和普通植物一样,在光下,更苏被子植物的光化学活性随着叶片失水而受到抑制,但奇怪的是在失去95％以上的水分后复水仍可迅速复活。在脱水过程中叶黄素循环和抗氧化系统的上调以及光合膜完整性和稳定性的保持,可能对更苏被子植物的耐脱水性起非常重要的作用。磷酸盐对复苏的影响也表现在复水阶段而且与上述两种保护机理关系不大,因此应该加强更苏被子植物复水阶段的研究。     

Desiccation-tolerant plants in dry environments

The majority of terrestrial plants are unable to survive in very dry environments. However, a small group of plants, called ‘resurrection’ plants, are extremely desiccation-tolerant and are capable of losing more than 90% of the cellular water in vegetative tissues. Resurrection plants can remain dried in an anabiotic state for several years and, upon rehydration, are able to resume normal growth and metabolism within 24 h. Vegetative desiccation tolerance is thought to have evolved independently several times within the plant kingdom from mechanisms that allow reproductive organs to survive air-dryness. Resurrection plants synthesise a range of compounds, either constitutively or in response to dehydration, that protect various components of the cell wall from damage during desiccation and/or rehydration. These include sugars and late embryogenesis abundant (LEA) proteins that are thought to act as osmoprotectants, and free radical-scavenging enzymes that limit the oxidative damage during dehydration. Changes in the cell wall composition during drying reduce the mechanical damage caused by the loss of water and the subsequent shrinking of the vacuole. These include an increase in expansin or cell wall-loosening activity during desiccation that enhances wall flexibility and promotes folding.