Hydraulic adjustment in jack pine and black spruce seedlings under controlled cycles of dehydration and rehydration

Drought adjustments were compared in black spruce ( Picea mariana [Mill] B.S.P), and jack pine ( Pinus banksiana [Lamb.]) by subjecting seedlings to five cycles of dehydration and rehydration. A computer-controlled root misting chamber system, supplied low (−1.5 MPa), moderate (−2.0 MPa), and severe (−2.5 MPa) dehydration, respectively, in cycles 1, 3 and 5. Although cell water relations failed to adjust to chronic dehydration, there was limited osmotic adjustment in black spruce (cycle 3), and water was re-allocated from the apoplast to the symplast in jack pine (cycles 1 and 3). Dehydration postponement was more important than dehydration tolerance. Jack pine was better able to postpone dehydration than black spruce. Specific conductivity, the hydraulic conductivity per unit stem cross-sectional area, was lower in jack pine and slower to decline during chronic dehydration. When specific conductivity was corrected for the greater leaf area in black spruce, the leaf-specific conductivity did not differ in the two species. There was no increase in needle leakage in jack pine and stomata in jack pine seedlings reopened fully after rehydration. Black spruce was more of a 'water spender', and less water stress (−2.0 MPa, cycle 3) was required to lower specific conductivity, compared to jack pine (−2.5 MPa, cycle 5). Leakage from needle membranes increased in black spruce, and stomata failed to reopen after rewatering (cycles 3 and 5). A greater needle area, smaller root system, and a higher specific conductivity lowered the water stress threshold for cavitation in black spruce, which is confined to moister sites in the boreal forest. Jack pine had a larger root system, smaller needle area and lower specific conductivity than black spruce. Because of these static features, jack pine is more drought tolerant and it is often found on sites that are too hot and dry for black spruce.     

The signature of seeds in resurrection plants: a molecular and physiological comparison of desiccation tolerance in seeds and vegetative tissues

Desiccation-tolerance in vegetative tissues of angiosperms hasa polyphyletic origin and could be due to 1) appropriation ofthe seed-specific program of gene expression that protects orthodoxseeds against desiccation, and/or 2) a sustainable version ofthe abiotic stress response. We tested these hypotheses by comparingmolecular and physiological data from the development of orthodoxseeds, the response of desiccation-sensitive plants to abioticstress, and the response of desiccation-tolerant plants to extremewater loss. Analysis of publicly-available gene expression dataof 35 LEA proteins and 68 anti-oxidant enzymes in the desiccation-sensitiveArabidopsis thaliana identified 13 LEAs and 4 anti-oxidantsexclusively expressed in seeds. Two (a LEA6 and 1-cys-peroxiredoxin)are not expressed in vegetative tissues in A. thaliana, buthave orthologues that are specifically activated in desiccatingleaves of Xerophyta humilis. A comparison of antioxidant enzymeactivity in two desiccation-sensitive species of Eragrostiswith the desiccation-tolerant E. nindensis showed equivalentresponses upon initial dehydration, but activity was retainedat low water content in E. nindensis only. We propose that theseantioxidants are housekeeping enzymes and that they are protectedfrom damage in the desiccation-tolerant species. Sucrose isconsidered an important protectant against desiccation in orthodoxseeds, and we show that sucrose accumulates in drying leavesof E. nindensis, but not in the desiccation-sensitive Eragrostisspecies. The activation of "seed-specific" desiccation protectionmechanisms (sucrose accumulation and expression of LEA6 and1-cys-peroxiredoxin genes) in the vegetative tissues of desiccation-tolerantplants points towards acquisition of desiccation tolerance fromseeds.     

Insights into the cellular mechanisms of desiccation tolerance among angiosperm resurrection plant species

Water is a major limiting factor in growth and reproduction in plants. The ability of tissues to survive desiccation is commonly found in seeds or pollen but rarely present in vegetative tissues. Resurrection plants are remarkable as they can tolerate almost complete water loss from their vegetative tissues such as leaves and roots. Metabolism is shut down as they dehydrate and the plants become apparently lifeless. Upon rehydration these plants recover full metabolic competence and ‘resurrect’. In order to cope with desiccation, resurrection plants have to overcome a number of stresses as water is lost from the cells, among them oxidative stress, destabilization or loss of membrane integrity and mechanical stress. This review will mainly focus on the effect of dehydration in angiosperm resurrection plants and some of the strategies developed by these plants to tolerate desiccation. Resurrection plants are important experimental models and understanding the physiological and molecular aspects of their desiccation tolerance is of great interest for developing drought‐tolerant crop species adapted to semi‐arid areas.     

The state of water in acclimating vegetative buds from Malus and Amelanchier and its relationship to winter hardiness

The relationship between freezable water and cold hardiness during acclimation was studied using vegetative buds from several apple ( Malus domestica Borkh) cultivars and from one saskatoonberry ( Amelanchier alnifolia Nutt. cv. Smoky) cultivar. According to leakage data and visual assessments of cortical browning, vegetative buds of all cultivars were most tolerant to subfreezing temperatures in January. The hardy condition was also associated with maximum tolerance to desiccation. Qualitative features of freezing exotherms (number of peaks and temperature of the transition) were not correlated with the hardy condition in the tissues. However, the amount of unfrozen water, determined by quantifying the energy of the exotherms, increased with increasing hardiness. In buds that survived exposure to −45°C, freezing reduced the intracellular water content, but only to levels above the critical moisture content for desiccation damage. In buds that did not survive exposure to −45°C, freezing reduced the water content to levels equal to or less than the critical moisture content for desiccation damage. These observations suggest that the freezing of water in nonhardy tissue dried the tissue to moisture levels at which severe dehydration damage occurred. It appears that acclimation of vegetative apple buds involves at least two processes: (1) an increase in tolerance to dehydration and (2) an increase in the level of unfreezable water.     

Sugar ratios, glutathione redox status and phenols in the resurrection species Haberlea rhodopensis and the closely related non-resurrection species Chirita eberhardtii

Because of their unique tolerance to desiccation, the so‐called resurrection plants can be considered as excellent models for extensive research on plant reactions to environmental stresses. The vegetative tissues of these species are able to withstand long dry periods and to recover very rapidly upon re‐watering. This study follows the dynamics of key components involved in leaf tissue antioxidant systems under desiccation in the resurrection plant Haberlea rhodopensis and the related non‐resurrection species Chirita eberhardtii. In H. rhodopensis these parameters were also followed during recovery after full drying. A well‐defined test system was developed to characterise the different responses of the two species under drought stress. Results show that levels of H₂O₂ decreased significantly both in H. rhodopensis and C. eberhardtii, but that accumulation of malondialdehyde was much more pronounced in the desiccation‐tolerant H. rhodopensis than in the non‐resurrection C. eberhardtii. A putative protective role could be attributed to accumulation of total phenols in H. rhodopensis during the late stages of drying. The total glutathione concentration and GSSG/GSH ratio increased upon complete dehydration of H. rhodopensis. Our data on soluble sugars suggest that sugar ratios might be important for plant desiccation tolerance. An array of different adaptations could thus be responsible for the resurrection phenotype of H. rhodopensis.     

Gene structure and expression analysis of the drought- and abscisic acid-responsive CDeT11-24 gene family from the resurrection plant Craterostigma plantagineum Hochst

In order to understand the molecular mechanisms which are responsible for desiccation tolerance in the resurrection plant Craterostigma plantagineum Hochst. a thorough analysis of the CDeT11-24 gene family was performed. CDeT11-24 comprises a small gene family whose genes are expressed in response to dehydration, salt stress and abscisic acid (ABA) treatment in leaves. The gene products are constitutively expressed in roots and disappear only when the plants are transferred to water. It is therefore suggested that the proteins are involved in sensing water status. The predicted proteins are very hydrophilic; they share some features with late-embryogenesis-abundant proteins, and sequence similarities were found with two ABA- and drought-regulated Arabidopsis genes. The analysis of β-glucuronidase reporter genes driven by the CDeT11-24 promoter showed high activity in mature seeds in both transgenic Arabidopsis and tobacco. In vegetative tissues the promoter activity in response to ABA was restricted to young Arabidosis seedlings. The responsiveness to ABA during later developmental stages was regained in the presence of the Arabidopsis gene product ABI3. Dehydration-induced promoter activity was only observed in Arabidopsis leaves at a particular developmental stage. This analysis indicates that some components in the signal transduction pathway of the resurrection plant are not active in tobacco or Arabidopsis. Received: 26 April 1997 / Accepted: 16 July 1997     

脱水导致的胞内溶质变化与植物耐干性的获得

植物耐干性是指许多植物个体和部分植物种子能在含水量极低的条件下存活,在回水的过程中迅速启动修复机制,细胞经重新水合修复所受损伤的能力。在脱水过程中,植物会合成和积累某些小分子物质、碳水化合物和特殊的蛋白质;在极度脱水状态下,多组分参与的玻璃化的形成和两性物质的重新分配、耐干性植物中特有的抗氧化机制都是植物获得耐干性的重要条件。复苏植物(resurrection plant)和部分被子植物种子是当前研究植物耐干性的模式材料。     

Freezing and desiccation tolerance of imbibed canola seed

Depending on the environmental conditions, imbibed seeds survive subzero temperatures either by supercooling or by tolerating freezing-induced desiccation. We investigated what the predominant survival mechanism is in freezing canola ( Brassica napus cv. Quest) and concluded that it depends on the cooling rate. Seeds cooled at 3°C h⁻¹ or faster supercooled, whereas seeds cooled over a 4-day period to −12°C and then cooled at 3°C h⁻¹ to−40°C did not display low temperature exotherms. Both differential thermal analysis and nuclear magnetic resonance (NMR) spectroscopy confirmed that imbibed canola seeds undergo freezing-induced desiccation at slow cooling rates. The freezing tolerance of imbibed canola seed (LT₅₀) was determined by slowly cooling to −12°C for 48 h, followed with cooling at 3°C h⁻¹ to −40°C, or by holding at a constant −6°C (LD₅₀). For both tests, the loss in freezing tolerance of imbibed seeds was a function of time and temperature of imbibition. Freezing tolerance was rapidly lost after radicle emergence. Seeds imbibed in 100 μ M abscisic acid (ABA), particularly at 2°C, lost freezing tolerance at a slower rate compared with water-imbibed seeds. Seeds imbibed in water either at 23°C for 16 h, or 8°C for 6 days, or 2°C for 6 days were not germinable after storage at −6°C for 10 days. Seeds imbibed in ABA at 23°C for 24 h, or 8°C for 8 days, or 2°C for 15 days were highly germinable after 40 days at a constant −6°C. Desiccation injury induced at a high temperature (60°C), as with injury induced by freezing, was found to be a function of imbibition temperature and time.     

Molecular characterization of two alanine-rich Lea genes abundantly expressed in the resurrection plant C. plantagineum in response to osmotic stress and ABA

Expression of many genes is induced during dehydration in vegetative tissues of the desiccation tolerant resurrection plantCraterostigma plantagineum. The most abundant group of desiccation-related gene products belong to the LEA (= Late Embryogenesis Abundant) proteins. Here we describe structures and expression patterns of members of group 3 and group 4Lea genes fromC. plantagineum. The most intriguing observation is the strong conservation of repeat motifs inLea genes found across divers plant species includingC. plantagineum and non-desiccation tolerant plants. This conservation of structural elements leads to speculations about evolution of desiccation tolerance in the resurrection plant.     

Desiccation tolerance in Physcomitrella patens: Rate of dehydration and the involvement of endogenous abscisic acid (ABA)

The moss Physcomitrella patens , a model system for basal land plants, tolerates several abiotic stresses, including dehydration. We previously reported that Physcomitrella patens survives equilibrium dehydration to ?13 MPa in a closed system at 91% RH. Tolerance of desiccation to water potentials below ?100 MPa was only achieved by pretreatment with exogenous abscisic acid (ABA). We report here that gametophores, but not protonemata, can survive desiccation below ?100 MPa after a gradual drying regime in an open system, without exogenous ABA. In contrast, faster equilibrium drying at 90% RH for 3–5 days did not induce desiccation tolerance in either tissue. Endogenous ABA accumulated in protonemata and gametophores under both drying regimes, so did not correlate directly with desiccation tolerance. Gametophores of a Ppabi3a/b/c triple knock out transgenic line also survived the gradual dehydration regime, despite impaired ABA signaling. Our results suggest that the initial drying rate, and not the amount of endogenous ABA, may be critical in the acquisition of desiccation tolerance. Results from this work will provide insight into ongoing studies to uncover the role of ABA in the dehydration response and the underlying mechanisms of desiccation tolerance in this bryophyte.     

Influence of germination date on Dioon edule (Zamiaceae) seedling tolerance to water stress

Dioon edule seedling mortality is mostly attributed to dehydration by prolonged drought, even when they present xeromorphic characteristics like the adult plants. The effect of germination date (GD) and soil water deficit on seedling tolerance to water stress was assessed. The seedlings germinated and grown from mature seeds every month from December to April GD were selected to evaluate the leaf area, photosynthetic pigment content, crassulacean acid metabolism (CAM) activity, stomatal conductance (gs) and leaflet anatomy at soil water potential (Ψs) of 0.0 MPa (day 1), ?0.1 MPa (day 40), ?1.0 MPa (day 90), ?1.5 MPa (day 130), and a control (0.0 MPa at day 130) to recognize differences due to leaf development. The seedlings shifted from C₃ to CAM cycling when exposed to water stress at Ψs of ?1.0 MPa, like adult plants. The March–April GD seedlings with undeveloped sclerified hypodermis and stomata, presented reduced leaf area, lower Chlorophyll a/b ratio, higher CAM activity and midday partial stomatal closure when reached Ψs of ?1.0 MPa. These have higher probability of dehydration during severe drought (February–April) than those of the December–February GD with similar Ψs. Plants used for restoration purposes must have full leaf development to increase the survival.     

A Putative LEA Protein,but no Trehalose,is Present in Anhydrobiotic Bdelloid Rotifers

Some eukaryotes, including bdelloid rotifer species, are able to withstand desiccation by entering a state of suspended animation. In this ametabolic condition, known as anhydrobiosis, they can remain viable for extended periods, perhaps decades, but resume normal activities on rehydration. Anhydrobiosis is thought to require accumulation of the non-reducing disaccharides trehalose (in animals and fungi) or sucrose (in plant seeds and resurrection plants), which may protect proteins and membranes by acting as water replacement molecules and vitrifying agents. However, in clone cultures of bdelloid rotifers Philodina roseola and Adineta vaga, we were unable to detect trehalose or other disaccharides in either control or dehydrating animals, as determined by gas chromatography. Indeed, trehalose synthase genes (tps) were not detected in these rotifer genomes, suggesting that bdelloids might not have the capacity to produce trehalose under any circumstances. This is in sharp contrast to other anhydrobiotic animals such as nematodes and brine shrimp cysts, where trehalose is present during desiccation. Instead, we suggest that adaptations involving proteins might be more important than those involving small biochemicals in rotifer anhydrobiosis: on dehydration, P. roseola upregulates a hydrophilic protein related to the late embryogenesis abundant (LEA) proteins associated with desiccation tolerance in plants. Since LEA-like proteins have also been implicated in the desiccation tolerance of nematodes and micro-organisms, it seems that hydrophilic protein biosynthesis represents a common element of anhydrobiosis across several biological kingdoms.     

Desiccation tolerant plants as model systems to study redox regulation of protein thiols

While the majority of plants and animals succumb to water loss, desiccation tolerant organisms can lose almost all of their intracellular water and revive upon rehydration. Only about 300 ‘resurrection’ angiosperms and very few animals are desiccation tolerant. By contrast, many bryophytes and most lichens are desiccation tolerant and so are the seeds and pollen grains of most flowering plants. The current literature reveals that the extreme fluctuations in water content experienced by desiccation tolerant organisms are accompanied by equally extreme changes in cellular redox state. Strongly oxidizing conditions upon desiccation can cause irreversible oxidation of free cysteine residues of proteins, which can change protein structure and function, and contribute to protein denaturation. It appears likely that reversible formation of disulphide bonds, in particular through protein glutathionylation, contributes to the set of protection mechanisms that confer desiccation tolerance. Upon rehydration, de-glutathionylation can be catalyzed by glutaredoxins (GRXs) and protein disulphide bonds can be reduced through NADPH-dependent thioredoxins (TRXs). Due to their ability to survive severe oxidative stress, desiccation tolerant plants and seeds are excellent models to study protein redox regulation, which may provide tools for enhancing tolerance to drought and more generally, to oxidative stress, in crops.     

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.