Pea seed mitochondria are endowed with a remarkable tolerance to extreme physiological temperatures

Most seeds are anhydrobiotes, relying on an array of protective and repair mechanisms, and seed mitochondria have previously been shown to harbor stress proteins probably involved in desiccation tolerance. Since temperature stress is a major issue for germinating seeds, the temperature response of pea (Pisum sativum) seed mitochondria was examined in comparison with that of mitochondria from etiolated epicotyl, a desiccation-sensitive tissue. The functional analysis illustrated the remarkable temperature tolerance of seed mitochondria in response to both cold and heat stress. The mitochondria maintained a well-coupled respiration between -3.5 degrees C and 40 degrees C, while epicotyl mitochondria were not efficient below 0 degrees C and collapsed above 30 degrees C. Both mitochondria exhibited a similar Arrhenius break temperature at 7 degrees C, although they differed in phospholipid composition. Seed mitochondria had a lower phosphatidylethanolamine-to-phosphatidylcholine ratio, fewer unsaturated fatty acids, and appeared less susceptible to lipid peroxidation. They also accumulated large amounts of heat shock protein HSP22 and late-embryogenesis abundant protein PsLEAm. The combination of membrane composition and stress protein accumulation required for desiccation tolerance is expected to lead to an unusually wide temperature tolerance, contributing to the fitness of germinating seeds in adverse conditions. The unique oxidation of external NADH at low temperatures found with several types of mitochondria may play a central role in maintaining energy homeostasis during cold shock, a situation often encountered by sessile and ectothermic higher plants.     

Seed Storage Behavior and Seed Germination of Nine Species of Lauraceae from Yunnan,China

Seed banking following internationally agreed standards is an important way for preserving collections of wild plant species ex situ; but this method is not suitable for desiccation sensitive species. Lauraceae comprehends some of the dominant species in the evergreen broadleaved forest in the south of China and contains many species both of ecological and economical importance. However, study on seed biology such as germination and desiccation tolerance of this family is scarce. Seeds of 9 species from 5 genera of this family were collected and their dormancy status and germination requirement were studied; also their desiccation tolerance were determined using a modified 100 seed test. The results showed that seeds of Cinnamomum camphora probably have intermediate physiological dormancy; seeds of Actinodaphne forrestii, Actinodaphne obovata, Cinnamomum migao, Lindera metcalfiana var. dictyophylla, Lindera communis and Neolitsea polycarpa are non deep physiological dormant; Seeds of Cinnamomum burmannii and Phoebe glaucophylla may have no or negligible dormancy. All 9 species lost seed viability after desiccated to 286%-716% moisture content while still retained considerable viability with moisture content ranged from 1732% to 4487% after moist storage; thus seeds of the 9 species are all desiccation sensitive and can not be stored at the conventional seed bank conditions.     

Induction of Hsp70 by desiccation, ionising radiation and heat-shock in the eutardigrade Richtersius coronifer

The physiology and biochemistry behind the extreme tolerance to desiccation shown by the so-called anhydrobiotic animals represents an exciting challenge to biology. The current knowledge suggests that both carbohydrates and proteins are often involved in protecting the dry cell from damage, or in the repair of induced damage. Tardigrades belong to the most desiccation-tolerant multicellular organisms, but very little research has been reported on the biochemistry behind desiccation tolerance in this group. We quantified the induction of the heat-shock protein Hsp70, a very wide-spread stress protein, in response to desiccation, ionising radiation, and heating, in the anhydrobiotic tardigrade Richtersius coronifer using an immuno-westernblot method. Elevated levels of Hsp70 were recorded after treatment of both heat and ionising radiation, and also in rehydrated tardigrades after a period of desiccation. In contrast, tardigrades in the desiccated (dry) state had reduced Hsp70 levels compared to the non-treated control group. Our results suggest that Hsp70 may be involved in the physiological and biochemical system underlying desiccation (and radiation) tolerance in tardigrades, and that its role may be connected to repair processes after desiccation rather than to biochemical stabilization in the dry state.     

A mitochondrial late embryogenesis abundant protein stabilizes model membranes in the dry state

Late embryogenesis abundant (LEA) proteins are a highly diverse group of polypeptides expected to play important roles in desiccation tolerance of plant seeds. They are also found in other plant tissues and in some anhydrobotic invertebrates, fungi, protists and prokaryotes. The LEA protein LEAM accumulates in the matrix space of pea (Pisum sativum) mitochondria during late seed maturation. LEAM is an intrinsically disordered protein folding into amphipathic α-helix upon desiccation. This suggests that it could interact with the inner mitochondrial membrane, providing structural protection in dry seeds. Here, we have used Fourier-transform infrared and fluorescence spectroscopy to gain insight into the molecular details of interactions of LEAM with phospholipid bilayers in the dry state and their effects on liposome stability. LEAM interacted specifically with negatively charged phosphate groups in dry phospholipids, increasing fatty acyl chain mobility. This led to an enhanced stability of liposomes during drying and rehydration, but also upon freezing. Protection depended on phospholipid composition and was strongly enhanced in membranes containing the mitochondrial phospholipid cardiolipin. Collectively, the results provide strong evidence for a function of LEAM as a mitochondrial membrane protectant during desiccation and highlight the role of lipid composition in the interactions between LEA proteins and membranes.     

What affects the desiccation tolerance threshold of Brazilian Eugenia (Myrtaceae) seeds?

Desiccation sensitive (DS) seeds are shed at high water contents (WC) and metabolically active, but WC thresholds vary broadly among species even in the same genus. Eugenia is an important ecological genus that has high occurrence in several Brazilian morphoclimatic domains. In this study, we assessed seed desiccation tolerance of five Eugenia species collected in specific meteorological conditions. We reported the species geographical ranges and verified the rainfall and temperature of species sites in the year prior to seed collection. We also assessed initial WC, seed germination and vigor and seedling growth upon desiccation. Eugenia uniflora was the widest spread among the five species, while E. astringens was the most restricted. In this specific study, widespread species showed a higher WC threshold than restricted species. In the same way, the WC of fresh seeds was not correlated to the desiccation tolerance threshold. Seed desiccation tolerance was species dependent and correlated with the environmental status of seed collection sites. Wetter and warmer conditions were correlated to the E. uniflora higher DS threshold. Low rainfall and temperature corresponded to a lower desiccation sensitivity of E. astringens seeds. Seeds of the five species lost half viability between 0.44 and 0.25 g H₂O g DW^??1 and after 65–270 h of desiccation. Our results indicate that abiotic factors impact plant populations during the seed production season and can drive seed desiccation tolerance threshold and physiological behavior. These results should be taken into account in ex-situ plant conservation programs and tropical species management.

     

Inheritance of seed desiccation sensitivity in a coffee interspecific cross: evidence for polygenic determinism

The genetic determinism of seed desiccation sensitivity was studied using a cross between two coffee species exhibiting a large difference for this trait, Coffea pseudozanguebariae (tolerant) and C. liberica (sensitive). Throughout the whole study, seed desiccation tolerance was quantified both in terms of water content and water activity. Whatever the parameter used, the level of seed desiccation tolerance in F1 hybrids corresponded to that of the mid-parent, thus indicating an additive inheritance of seed desiccation tolerance at the F1 level. A broad variation was observed among hybrids backcrossed to C. liberica (BCs) for seed desiccation tolerance, independent of the parameter used to quantify it. This variation was continuous and BCs showed transgression in the direction of the most desiccation sensitive parent, indicating (i) that desiccation tolerance is a polygenic trait in coffee species, and (ii) that C. pseudozanguebariae does not present the most favourable alleles for all the genes involved in seed desiccation tolerance. No significant difference was observed between the two reciprocal backcrosses, F1xC. liberica and C. libericaxF1, for the level of desiccation tolerance of their seeds, showing the absence of a maternal effect on this trait. There was no significant effect of the number of seeds harvested from each BC on the level of desiccation tolerance of its seeds. Moreover, there was no significant correlation within BCs between seed size, seed viability, and water content before desiccation and desiccation tolerance.     

Nitric oxide enhances desiccation tolerance of recalcitrant Antiaris toxicaria seeds via protein S-nitrosylation and carbonylation

The viability of recalcitrant seeds is lost following stress from either drying or freezing. Reactive oxygen species (ROS) resulting from uncontrolled metabolic activity are likely responsible for seed sensitivity to drying. Nitric oxide (NO) and the ascorbate-glutathione cycle can be used for the detoxification of ROS, but their roles in the seed response to desiccation remain poorly understood. Here, we report that desiccation induces rapid accumulation of H(2)O(2), which blocks recalcitrant Antiaris toxicaria seed germination; however, pretreatment with NO increases the activity of antioxidant ascorbate-glutathione pathway enzymes and metabolites, diminishes H(2)O(2) production and assuages the inhibitory effects of desiccation on seed germination. Desiccation increases the protein carbonylation levels and reduces protein S-nitrosylation of these antioxidant enzymes; these effects can be reversed with NO treatment. Antioxidant protein S-nitrosylation levels can be further increased by the application of S-nitrosoglutathione reductase inhibitors, which further enhances NO-induced seed germination rates after desiccation and reduces desiccation-induced H(2)O(2) accumulation. These findings suggest that NO reinforces recalcitrant seed desiccation tolerance by regulating antioxidant enzyme activities to stabilize H(2)O(2) accumulation at an appropriate concentration. During this process, protein carbonylation and S-nitrosylation patterns are used as a specific molecular switch to control antioxidant enzyme activities.     

与种子耐脱水性有关的基础物质研究进展

耐脱水的获得和维持与种子的类型有关,正常型种子耐脱水,而顽拗形种子对脱水高度敏感,正常型种子的脱水耐性随发良[过程而变化,种子成熟时胚的脱水耐性增强,其萌发时胚变为不耐脱水,当种子获得脱水耐性时,糖,LEA蛋白质和抗氧化防御系统等保护性物质积累,但脱水耐性是一种复杂的数量的特性,任何一种单一的机制都不能 充分地解释脱水耐性,各种保护性物质协同调节脱水耐性,本文综述了近几年来关于种子耐脱水性与保护性物质相关性的研究进展。     

Modelling the effects of microclimate on bean seed desiccation rate and seed storage ability

Bean seed storage ability is of major interest for seed firms and depends, at least partially, on the effect of desiccation microclimate and desiccation rate on the mother plant; however, their effects are contradictory and not quantified. Therefore, our aims were (a) to predict seed desiccation rate from microclimatic variables at the pod level and (b) to measure the effects of seed desiccation rate and temperature on seed storage ability. For 4 years, beans (Phaseolus vulgaris) were sown at different dates in the field and once in a greenhouse. Pods having seeds at the beginning of desiccation stage were selected according to their colour and then dried in the field or under controlled conditions. In the field, pods were dried under both natural conditions and conditions modified either by defoliation or by water spraying. Under controlled conditions, pods were dried under a wide range of temperature and relative humidity combinations. Seed water content, temperature and relative humidity were measured throughout the desiccation phase. Storage ability was measured by a standard germination test after either 11 or 15 days of controlled deterioration. Seed desiccation rate was highly correlated to air vapour pressure deficit, which measures the combined effects of temperature and relative humidity on drying. With a logit transformation, deterioration test results showed a linear decrease in storage ability as seed desiccation rate increased for either controlled or field‐dried lots. However, for controlled lots desiccated at temperatures lower than 24°C, the lower the temperature, the lower the storage ability at a given desiccation rate. Moreover, the relationship between the results of the two deterioration periods differed between controlled and field‐dried lots. Finally, the high correlation between seed storage ability, desiccation rate and vapour pressure deficit in the field has given new insights for building a crop decision‐oriented model for optimising harvesting.     

Tree seed traits’ response to monsoon climate and altitude in Indian subcontinent with particular reference to the Himalayas

Seed traits are related to several ecological attributes of a plant species, including its distribution. While the storage physiology of desiccation‐sensitive seeds has drawn considerable attention, their ecology has remained sidelined, particularly how the strong seasonality of precipitation in monsoonal climate affects their temporal and spatial distribution. We compiled data on seed mass, seed desiccation behavior, seed shedding, and germination periodicity in relation to monsoon and altitude for 198 native tree species of Indian Himalayas and adjoining plains to find out (1) the adaptive significance of seed mass and seed desiccation behavior in relation to monsoon and (2) the pattern of change in seed mass in relation to altitude, habitat moisture, and succession. The tree species fall into three categories with respect to seed shedding and germination periodicities: (1) species in which both seed shedding and germination are synchronized with monsoon, referred to as monsoon‐synchronized (MS, 46 species); (2) species in which seed germination is synchronized with monsoon, but seeds are shed several months before monsoon, referred to as partially monsoon‐synchronized (PMS, 112 species); and (3) species in which both shedding and germination occur outside of monsoon months, referred to as monsoon‐desynchronized (MD, 39 species). The seed mass of MS species (1,718 mg/seed) was greater than that of PMS (627 mg/seed) and MD (1,144 mg/seed). Of the 40 species with desiccation‐sensitive seeds, 45% belong to the MS category, almost similar (approx. 47%) to woody plants with desiccation‐sensitive seeds in evergreen rain forests. Seed mass differed significantly as per seed desiccation behavior and successional stage. No relationship of seed mass was found with altitude alone and on the basis of seed desiccation behavior. However, seed mass trend along the altitude differed among monsoon synchronization strategies. Based on our findings, we conclude that in the predicted climate change (warming and uncertain precipitation pattern) scenario, a delay or prolonged break‐spell of monsoon may adversely affect the regeneration ecology of desiccation‐sensitive seed‐bearing species dominant over large forest areas of monsoonal climate.     

Desiccation tolerance in developing soybean seeds: The role of stress proteins

The consistent correlation between desiccation tolerance in orthodox seed tissue and an accumulation of certain "late embryogenesis abundant" (LEA) proteins suggests that these proteins reduce desiccation-induced cellular damage. The aim of the present work was to test this hypothesis. Exogenous abscisic acid (ABA) was used to elevate the level of heal-soluble LEA-like proteins in axes from immature (30 days after flowering: mid-development) seeds of soybean ( Glycine max [L.] Merrill cv. Chippewa 64). As the LEA-like proteins accumulated in response to ABA, the leakage of all elements after desiccation and subsequent rehydration markedly declined. Both LEA-like protein accumulation and the decline in desiccation-induced electrolyte leakage were apparently dependent on the presence of ABA. Both effects of ABA were inhibited by cycloheximide. Light microscopy revealed a marked effect of the ABA on cellular integrity following desiccation. Osmotic stress also caused a decrease in desiccation-induced electrolyte leakage and stimulated the accumulation of LEA-like proteins. Our data are consistent with the hypothesis that the LEA-like proteins contribute to the increase in desiccation tolerance in response to ABA, and are consistent with a general protective role for these proteins in desiccation tolerance.     

Prediction of desiccation sensitivity in seeds of woody species: a probabilistic model based on two seed traits and 104 species

BACKGROUND AND AIMS: Seed desiccation sensitivity limits the ex situ conservation of up to 47 % of plant species, dependent on habitat. Whilst desirable, empirically determining desiccation tolerance levels in seeds of all species is unrealistic. A probabilistic model for the rapid identification of woody species at high risk of displaying seed desiccation sensitivity is presented. METHODS: The model was developed using binary logistic regression on seed trait data [seed mass, moisture content, seed coat ratio (SCR) and rainfall in the month of seed dispersal] for 104 species from 37 families from a semi-deciduous tropical forest in Panamá. KEY RESULTS: For the Panamanian species, only seed mass and SCR were significantly related to the response to desiccation, with the desiccation-sensitive seeds being large and having a relatively low SCR (i.e. thin 'seed' coats). Application of this model to a further 38 species, of known seed storage behaviour, from two additional continents and differing vegetation types (dryland Africa and temperate Europe) correctly predicted the response to desiccation in all cases, and resolved conflicting published data for two species (Acer pseudoplatanus and Azadirachta indica). CONCLUSIONS: This model may have application as a decision-making tool in the handling of species of unknown seed storage behaviour in species from three disparate habitats.     

Patterns of desiccation tolerance and recovery in bryophytes

The study of desiccation tolerance in bryophytes avoids thecomplications of higher-plant vascular systems and complex leaf structures, butremains a multifaceted problem. Some of the pertinent questions have at leastpartial analogues in seed biology – events during a drying-rewettingcyclewith processes in seed maturation and germination, and the gradual loss ofviability on prolonged desiccation, and the relation of this to intensity ofdesiccation and temperature, with parallel questions in seed storage. Pastresearch on bryophyte desiccation tolerance is briefly reviewed. Evidence ispresented from chlorophyll-fluorescence measurements and experiments withmetabolic inhibitors that recovery of photosynthesis in bryophytes followingdesiccation depends mainly on rapid reactivation of pre-existing structures andinvolves only limited de novo protein synthesis. Followinginitial recovery, protein synthesis is demonstrably essential to themaintenanceof photosynthetic function in the light, but the rate of maintenance turnoverinthe dark appears to be slow. Factors leading to long-term desiccation damagearediverse; indications are that desiccation tolerant species often survive bestinthe range –100 to –200 MPa.     

Gene expression during seed maturation in Brassica napus in relation to the induction of secondary dormancy

Gene expression in two cultivars of Brassica napus (AC Excel and DH12075) has been compared at the full-size embryo, desiccation, and mature stages of seed development. Seed of these cultivars differ in their potential to exhibit secondary dormancy following environmental stress; Excel has high potential and DH12075 has low potential. A majority of genes were down-regulated during maturation in both cultivars but a significant number of differences in gene expression between the cultivars were apparent in the transition from full-size embryo to mature seed. However, most differences were apparent in the desiccation stage and some of the differences were in genes related to signaling processes and protein biosynthesis. We suggest that the propensity of Brassica seeds to manifest secondary dormancy may be determined by changes in gene expression that occur during late seed development.     

Basis of coffee seed sensitivity to liquid nitrogen exposure: oxidative stress or imbibitional damage?

Two hypotheses, namely the occurrence of post‐thaw oxidative stress or imbibitional damage, were tested to explain the high sensitivity of coffee seeds to liquid nitrogen (LN) exposure. Oxidative stress was studied by measuring primary and secondary products of lipid peroxidation in seeds during the desiccation and rehydration periods. The 4‐hydroxynonenal (4‐HNE) content of seeds remained constant throughout the desiccation step. No significant difference was observed between desiccated seeds and seeds desiccated and exposed to LN for the evolution of their 4‐HNE and hydroperoxide contents during rehydration. In both cases, an increase in 4‐HNE and hydroperoxide contents of seeds was observed during the first hours of culture under germination conditions, followed by a progressive decrease down to values comparable to those observed in desiccated seeds. The hydroperoxide composition of frozen seeds was not significantly different from that of control seeds. The (S)/(R) enantiomeric ratios of 9‐ and 13‐hydroxy‐octadecadienoic acid extracted from rehydrating seeds were chiral, suggesting that they originated from lipoxygenase activity. These results suggest that the high sensitivity of coffee seeds to LN exposure is not directly associated with the occurrence of an oxidative stress during post‐thaw rehydration. The effect on seed viability of different rehydration procedures previously identified to reduce membrane imbibitional injury was studied after desiccation and LN exposure. Desiccation tolerance increased with, by increasing order, seed osmoconditioning, pre‐heating and pre‐humidifying prior to their culture under germination conditions. Among the four combinations of pre‐humidification durations (24 or 48 h) and temperatures (25 or 37°C) tested, pre‐humidification for 24 h at 37°C gave the highest level of desiccation tolerance. This rehydration procedure also dramatically increased seed viability after LN exposure. Seed desiccation sensitivity modelling in combination with the calculation of the decrease in seed water activity during cooling facilitated the explanation of the beneficial effect of controlled rehydration after desiccation and LN exposure. These results support the hypothesis that imbibitional membrane damage is involved in the sensitivity of coffee seeds to LN exposure.     

Temporal profiling of the heat-stable proteome during late maturation of Medicago truncatula seeds identifies a restricted subset of late embryogenesis abundant proteins associated with longevity

Developing seeds accumulate late embryogenesis abundant (LEA) proteins, a family of intrinsically disordered and hydrophilic proteins that confer cellular protection upon stress. Many different LEA proteins exist in seeds, but their relative contribution to seed desiccation tolerance or longevity (duration of survival) is not yet investigated. To address this, a reference map of LEA proteins was established by proteomics on a hydrophilic protein fraction from mature Medicago truncatula seeds and identified 35 polypeptides encoded by 16 LEA genes. Spatial and temporal expression profiles of the LEA polypeptides were obtained during the long maturation phase during which desiccation tolerance and longevity are sequentially acquired until pod abscission and final maturation drying occurs. Five LEA polypeptides, representing 6% of the total LEA intensity, accumulated upon acquisition of desiccation tolerance. The gradual 30-fold increase in longevity correlated with the accumulation of four LEA polypeptides, representing 35% of LEA in mature seeds, and with two chaperone-related polypeptides. The majority of LEA polypeptides increased around pod abscission during final maturation drying. The differential accumulation profiles of the LEA polypeptides suggest different roles in seed physiology, with a small subset of LEA and other proteins with chaperone-like functions correlating with desiccation tolerance and longevity.     

Trehalose loading through the mitochondrial permeability transition pore enhances desiccation tolerance in rat liver mitochondria

Trehalose has extensively been used to improve the desiccation tolerance of mammalian cells. To test whether trehalose improves desiccation tolerance of mammalian mitochondria, we introduced trehalose into the matrix of isolated rat liver mitochondria by reversibly permeabilizing the inner membrane using the mitochondrial permeability transition pore (MPTP). Measurement of the trehalose concentration inside mitochondria using high performance liquid chromatography showed that the sugar permeated rapidly into the matrix upon opening the MPTP. The concentration of intra-matrix trehalose reached 0.29 mmol/mg protein (∼190 mM) in 5 min. Mitochondria, with and without trehalose loaded into the matrix, were desiccated in a buffer containing 0.25 M trehalose by diffusive drying. After re-hydration, the inner membrane integrity was assessed by measurement of mitochondrial membrane potential with the fluorescent probe JC-1. The results showed that following drying to similar water contents, the mitochondria loaded with trehalose had significantly higher inner membrane integrity than those without trehalose loading. These findings suggest the presence of trehalose in the mitochondrial matrix affords improved desiccation tolerance to the isolated mitochondria.     

Disappearance of desiccation tolerance of imbibed crop seeds is not associated with the decline of oligosaccharides

The relationship between sugar content and loss of desiccation tolerance of hydrated crop seeds (tomato, okra, snow pea, mung bean, and cucumber) was evaluated by imbibing seeds with or without ABA, followed by dehydration and germination. During the process of hydration, but before the seeds lost desiccation tolerance, monosaccharide content increased only slightly, sucrose increased in snow peas, mung bean and cucumber, but maintained its original level in other species and the oligosaccharides declined dramatically. At the time of losing desiccation tolerance, the sucrose content of imbibed seeds was 2-3 times higher than the original level in most species. Positive significant correlation coefficients (r) were found in many, but not all crop seeds between desiccation tolerance and the oligosaccharide mass, or oligo/sucrose ratio. The ratio of oligo/sucrose in intact seeds at the time of losing desiccation tolerance, however, was not a fixed value and varied among species. Oligosaccharides declined significantly in different seed parts of imbibed cucumber seeds while sucrose increased to a higher level in the radicle than in the hypocotyl. Radicles were far more sensitive to desiccation than hypocotyls. The same observation was found for cucumber seeds imbibed in 100 M ABA, yet desiccation tolerance was largely maintained in hypocotyls and cotyledons. It is concluded that sucrose and oligosaccharides are not the determinants of the loss of desiccation tolerance in hydrated seeds.Imbibed seeds did not show any differences between seed parts in their ability to resynthesize sugars during the process of slow dehydration. Differences in sensitivity to desiccation among seed parts were not due to differences in the initial water content or to the rate of water content increase among seed parts. Physiological regulation of the loss of desiccation tolerance in crop seeds during hydration is discussed.