Microsatellite Polymorphism and the Population Structure of the Black Tiger Shrimp (Penaeus monodon) in Thailand

: Genetic variation and differentiation of Thai Penaeus monodon from five geographic locations (Chumphon, Trad, Phangnga, Satun, and Trang) were investigated using five microsatellite loci (CUPmo18, Di25, Di27, CSCUPmo1, and CSCUPmo2). The number of alleles across the five loci ranged from 19 to 30, and heterozygosities ranged from 0.49 to 0.95. The mean number of alleles and effective number of alleles per locus were 21.0 to 26.6 and 13.1 to 20.4, respectively. The average heterozygosity across all investigated samples was 0.78, indicating high genetic diversity in this species. Geographic heterogeneity analysis of the results from two of the loci, CUPmo18 and Di25, showed significant differences among the Gulf of Thailand (Trad and Chumphon) but not the Andaman samples. Comparison between regions revealed significant heterogeneity of the Andaman and Trad P. monodon (P < .001), whereas those from Chumphon and the Andaman were genetically similar (P > .05). Significant genetic differentiation was consistently observed between the Andaman-Trad samples (FST = 0.0101, P < .0001) and the Chumphon-Trad samples (FST = 0.0101, P < .0001). On the basis of our analyses, the investigated samples from five geographic locations were allocated to three distinct populations composed of the Andaman Sea (A), Chumphon (B), and Trad (C).     

Citrate synthase from Antarctic ciliates: adaptation to low temperatures and comparison with temperate ciliates

The activity of citrate synthase (CS), a mitochondrial enzyme in the energy metabolism pathway, was studied in Euplotes focardii (Hypotrichia) and Uronema marinum (Scuticociliatia), isolated from samples of Antarctic seawater and, for comparison in E. vannus and U. nigricans, related ciliates from the Adriatic. The temperature-dependent characteristics of CS were investigated in the range 0–55°C, to evaluate mechanisms of thermal regulation, such as optimal temperature levels, activation energy, and kinetic parameters. CS activity was up to double that recorded in temperate organisms at reaction temperatures between 0 and 10°C. The optimal temperature for enzymatic activity was independent of Tamb. Activation energy for CS was significantly lower in Antarctic ciliates than in temperate ones, indicating a significant increase in the reaction rate. KM at various temperatures of the reaction substrates indicated the higher affinity of CS for acetyl-CoA in both Antarctic organisms at lower temperatures. The data for thermal profiles for KM, showing greater differences between E. focardii and E. vannus than between U. marinum and U. nigricans, support the distinctive physiological characteristics of each species. E. focardii is an endemic and psychrophilic species, whereas U. marinum is a typically ubiquitous species, more adaptable to environmental changes.     

Characterization of singlet oxygen-accumulating mutants isolated in a screen for altered oxidative stress response in <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>

Background  

When photosynthetic organisms are exposed to harsh environmental conditions such as high light intensities or cold stress, the production of reactive oxygen species like singlet oxygen is stimulated in the chloroplast. In Chlamydomonas reinhardtii singlet oxygen was shown to act as a specific signal inducing the expression of the nuclear glutathione peroxidase gene GPXH/GPX5 during high light stress, but little is known about the cellular mechanisms involved in this response. To investigate components affecting singlet oxygen signaling in C. reinhardtii, a mutant screen was performed.     

A difference in optomotor behaviour of two Antarctic nototheniid fishes is correlated with the presence of a choroid<Emphasis Type="Italic"> rete mirabile</Emphasis> and Root effect

The hypothesis that fish vision is influenced by the presence of a choroid rete mirabile (an oxygen-concentrating apparatus in the eye) and the Root effect was tested using the optomotor response in two nototheniid species from Antarctica. Pagothenia borchgrevinki, an active cryopelagic zooplanktivore, does not exhibit a major Root effect and the rete is absent. In contrast, the sluggish, benthic nototheniid, Trematomus bernacchii, possesses both a rete and a significant Root effect. Whole-eye ocular PO2 was higher in T. bernacchii (18.6 kPa) than in P. borchgrevinki (18.1 kPa) and the optomotor response of T. bernacchii was positively influenced by subtended angle on a moving background whereas P. borchgrevinki did not respond to any subtended angle (0–600 min of arc). The optomotor response of T. bernacchii may therefore have been facilitated by elevated ocular PO2 due to the presence of the choroid rete and the Root effect.     

Bacillus naphthovorans sp. nov. from oil-contaminated tropical marine sediments and its role in naphthalene biodegradation

. A Bacillus sp., designated as strain MN-003, was isolated as the dominant cultivatable naphthalene-degrading organism from oil-contaminated tropical marine sediments. Strain MN-003 is strictly aerobic, rod-shaped, Gram-positive, catalase positive, oxidase negative, and forms endospores. Strain MN-003 grew at salinities ranging from 0.28 to 7.00% and temperatures ranging from 15 to 41°C. Phylogenetic analyses reveal that strain MN-003 is most similar to Bacillus sp. VAN14, with a 16S rRNA sequence identity of 97.9%. Based on taxonomic and 16S rRNA data, strain MN-003 was named Bacillus naphthovorans sp. nov. When grown with naphthalene as sole carbon source, strain MN-003 had a maximal specific growth rate (µmax) of 0.32ǂ.03 h^–1, and a half-saturation constant (Ks) of 22.3dž.2 µM. A batch study of the tropical marine sediments enriched with naphthalene showed that cells of the Bacillus genus grew to become dominant members of the microbial community. The bacilli comprised 39.5Lj.5% of the microbial fraction after 20 days of enrichment.     

A Mediator of Singlet Oxygen Responses in Chlamydomonas reinhardtii and Arabidopsis Identified by a Luciferase-Based Genetic Screen in Algal Cells

All cells produce reactive oxygen species (ROS) as by-products of their metabolism. In addition to being cytotoxic, ROS act as regulators of a wide range of developmental and physiological processes. Little is known about the molecular mechanisms underlying the perception of ROS and initiation of cellular responses in eukaryotes. Using the unicellular green alga Chlamydomonas reinhardtii, we developed a genetic screen for early components of singlet oxygen signaling. Here, we report the identification of a small zinc finger protein, METHYLENE BLUE SENSITIVITY (MBS), that is required for induction of singlet oxygen–dependent gene expression and, upon oxidative stress, accumulates in distinct granules in the cytosol. Loss-of-function mbs mutants produce singlet oxygen but are unable to fully respond to it at the level of gene expression. Knockout or knockdown of the homologous genes in the higher plant model Arabidopsis thaliana results in mutants that are hypersensitive to photooxidative stress, whereas overexpression produces plants with elevated stress tolerance. Together, our data indicate an important and evolutionarily conserved role of the MBS protein in ROS signaling and provide a strategy for engineering stress-tolerant plants.     

A comparative study of the hormonal response to high temperatures and stress reiteration in three Labiatae species

Mediterranean plants are usually exposed to a combination of stresses, which may occur simultaneously or at different times throughout their life. Here, the hormonal response to high temperatures was compared in plants of three Labiatae species, including rosemary (Rosmarinus officinalis L.), sage (Salvia officinalis L.) and lemon balm (Melissa officinalis L.). Plants exposed to increasing temperatures for 5 days were subsequently exposed to heat stress and compared to plants experiencing heat stress for the first time (controls). Despite the three species showing a similar tolerance to a single heat stress event, stress recurrence had a different impact on each species. Lemon balm was the most sensitive species to stress reiteration, showing decreased relative water content upon heat stress repetition, together with enhanced levels of α-tocopherol and salicylic acid (SA). Some acclimation responses were observed in rosemary and sage, including improved water contents and reduced jasmonic acid levels in rosemary, and reduced abscisic acid (ABA) and malondialdehyde (MDA) levels in sage plants previously exposed to high temperatures. Furthermore, the response of plants to a combination of heat stress and water deficit was evaluated in plants previously exposed to heat stress and compared to controls. Rosemary and sage were much more resistant than lemon balm, which died when stresses were combined. Despite stress pre-exposure not having any effect on plant performance in terms of F_v/F_m, MDA and relative water content in rosemary and sage, it resulted in higher α-tocopherol levels in both species. The hormonal response differed between species: while the hormonal content did not change in sage, rosemary showed increased ABA and decreased SA levels as a result of repeated stress exposure. Overall, different stress imprints particular to each species and stress scenario were found in α-tocopherol and hormone levels, which led to similar protective effects in rosemary and sage.     

Acclimation by suboptimal growth temperature diminishes photooxidative damage in maize leaves

Leaves of Zea mays L. seedlings which developed at optimal (25°C) or suboptimal (15°C) temperature were exposed to high irradiance (1000 μmol m^?2 s^?1) and a severe chilling temperature (5°C) for up to 24 h to investigate their ability to withstand photooxidative stress. During this stress, the degradation of the endogenous antioxidants ascorbate, glutathione and α-tocopherol was delayed and less pronounced in 15°C leaves. Similarly, the decline in chlorophyll a, chlorophyll b, β-carotene and lutein was slower throughout the stress period. Faster development and a higher level of non-photochemical quenching (NPQ) of chlorophyll fluorescence, related to a stronger de-poxidation of the larger xanthophyll cycle pool in 15°C leaves, could act as a defence mechanism to reduce the formation of reactive oxygen species during severe chilling. Furthermore, plants grown at suboptimal temperature exhibited a higher amount of the antioxidants glutathione and α-tocopherol. The higher α-tocopherol content in leaves (double based on leaf area; 4-fold higher based on chlorophyll content) which developed at suboptimal temperature may play an especially important role in the stabilization of the thylakoid membrane and thus prevent lipid peroxidation.     

Polyphenol oxidase-mediated protection against oxidative stress is not associated with enhanced photosynthetic efficiency

Background and Aims Polyphenol oxidases (PPOs) catalyse the oxidation of monophenols and/or o-diphenols to highly reactive o-quinones, which in turn interact with oxygen and proteins to form reactive oxygen species (ROS) and typical brown-pigmented complexes. Hence PPOs can affect local levels of oxygen and ROS. Although the currently known substrates are located in the vacuole, the enzyme is targeted to the thylakoid lumen, suggesting a role for PPOs in photosynthesis. The current study was designed to investigate the potential involvement of PPOs in the photosynthetic response to oxidative stress.Methods Photosynthesis (A, F_v/F_m, ΦPSII, q_N, q_P, NPQ) was measured in leaves of a wild-type and a low-PPO mutant of red clover (Trifolium pratense ‘Milvus’) under control conditions and under a stress treatment designed to induce photooxidative stress: cold/high light (2 °C/580 µmol m²s^–1) or 0–10 µm methyl viologen. Foliar protein content and oxidation state were also determined.Key Results Photosynthetic performance, and chlorophyll and protein content during 4 d of cold/high light stress and 3 d of subsequent recovery under control growth conditions showed similar susceptibility to stress in both lines. However, more extensive oxidative damage to protein in mutants than wild-types was observed after treatment of attached leaves with methyl viologen. In addition, PPO activity could be associated with an increased capacity to dissipate excess energy, but only at relatively low methyl viologen doses.Conclusions The presence of PPO activity in leaves did not correspond to a direct role for the enzyme in the regulation or protection of photosynthesis under cold stress. However, an indication that PPO could be involved in cellular protection against low-level oxidative stress requires further investigation.     

Control of the appearance of ascorbate peroxidase (EC 1.11.1.11) in mustard seedling cotyledons by phytochrome and photooxidative treatments

In photosynthetic cells the plastidic ascorbate-glutathione pathway is considered the major sequence involved in the elimination of active oxygen species. Ascorbate peroxidase (APO; EC 1.11.1.11) is an essential constituent of this pathway. In the present paper control of the appearance of APO was studied in the cotyledons of mustard (Sinapis alba L.) seedlings with the following results: (i) Two isoforms of APO (APO I, APO II) could be separated by anion-exchange chromatography; APO I is a plastidic protein, while APO II is extraplastidic, very probably cytosolic. (ii) The appearance of APO is regulated by light via phytochrome. This control is observed with both isoforms. Moreover, a strong positive control over APO II appearance (very probably over APO II synthesis) is exerted by photooxidative treatment of the plastids. (iii) Additional synthesis of extraplastidic APO II is induced by a signal created by intraplastidic pigment-photosensitized oxidative stress. The response is obligatorily oxygen-dependent and abolished by quenchers of singlet oxygen such as -tocopherol and p-benzoquinone. (iv) A short-term (4 h) photooxidative treatment suffices to saturate the signal. Signal transduction cannot be abolished or diminished by replacing the plants in non-photooxidizing conditions. Several observations indicate that control of APO synthesis by active oxygen is not an experimental artifact but a natural phenomenon.Abbreviations APO ascorbate-specific peroxidase (EC 1.11.1.11) - D darkness - FPLC fast protein liquid chromatography - FR far-red light (3.5 W · m^–2) - NF Norflurazon - R red light (6.8 W · m^–2) This research was supported by a grant from the Deutsche For-schungsgemeinschaft. B. Th. was the recipient of a stipend from the Studienstiftung des Deutschen Volkes.     

Different roles of alpha- and beta-branch xanthophylls in photosystem assembly and photoprotection

Xanthophylls (oxygenated carotenoids) are essential components of the plant photosynthetic apparatus, where they act in photosystem assembly, light harvesting, and photoprotection. Nevertheless, the specific function of individual xanthophyll species awaits complete elucidation. In this work, we analyze the photosynthetic phenotypes of two newly isolated Arabidopsis mutants in carotenoid biosynthesis containing exclusively alpha-branch (chy1chy2lut5) or beta-branch (chy1chy2lut2) xanthophylls. Both mutants show complete lack of qE, the rapidly reversible component of nonphotochemical quenching, and high levels of photoinhibition and lipid peroxidation under photooxidative stress. Both mutants are much more photosensitive than npq1lut2, which contains high levels of viola- and neoxanthin and a higher stoichiometry of light-harvesting proteins with respect to photosystem II core complexes, suggesting that the content in light-harvesting complexes plays an important role in photoprotection. In addition, chy1chy2lut5, which has lutein as the only xanthophyll, shows unprecedented photosensitivity even in low light conditions, reduced electron transport rate, enhanced photobleaching of isolated LHCII complexes, and a selective loss of CP26 with respect to chy1chy2lut2, highlighting a specific role of beta-branch xanthophylls in photoprotection and in qE mechanism. The stronger photosystem II photoinhibition of both mutants correlates with the higher rate of singlet oxygen production from thylakoids and isolated light-harvesting complexes, whereas carotenoid composition of photosystem II core complex was not influential. In depth analysis of the mutant phenotypes suggests that alpha-branch (lutein) and beta-branch (zeaxanthin, violaxanthin, and neoxanthin) xanthophylls have distinct and complementary roles in antenna protein assembly and in the mechanisms of photoprotection.     

The Function of Tocopherols and Tocotrienols in Plants

Referee: Dr. Kozi Asada, Department of Biotechnology, Faculty of Engineering, Fukuyama University, Gakuencho 1, Fukuyama 729-0292, Japan Tocopherols and tocotrienols, which differ only in the degree of saturation of their hydrophobic prenyl side chains, are lipid-soluble molecules that have a number of functions in plants. Synthesized from homogentisic acid and isopentenyl diphosphate in the plastid envelope, tocopherols and tocotrienols are essential to maintain membrane integrity. α-Tocopherol is the major form found in green parts of plants, while tocotrienols are mostly found in seeds. These compounds are antioxidants, thus they protect the plant from oxygen toxicity. Tocopherols and tocotrienols scavenge lipid peroxy radicals, thereby preventing the propagation of lipid peroxidation in membranes, and the ensuing products tocopheroxyl and tocotrienoxyl radicals, respectively, are recycled back to tocopherols and tocotrienols by the concerted action of other antioxidants. Furthermore, tocopherols and tocotrienols protect lipids and other membrane components by physically quenching and reacting chemically with singlet oxygen. The scavenging of singlet oxygen by α-tocopherol in chloroplasts results in the formation of, among other products, α -tocopherol quinone, a known contributor to cyclic electron transport in thylakoid membranes, therefore providing photoprotection for chloroplasts. Moreover, given that α-tocopherol increases membrane rigidity, its concentration, together with that of the other membrane components, might be regulated to afford adequate fluidity for membrane function. Furthermore, α-tocopherol may affect intracellular signaling in plant cells. The effects of this compound in intracellular signaling may be either direct, by interacting with key components of the signaling cascade, or indirect, through the prevention of lipid peroxidation or the scavenging of singlet oxygen. In the latter case, α-tocopherol may regulate the concentration of reactive oxygen species and plant hormones, such as jasmonic acid, within the cell, which control both the growth and development of plants, and also plant response to stress.     

How do Mediterranean shrub species cope with shade? Ecophysiological response to different light intensities

• Under natural conditions, light exposure for Mediterranean shrubs can be highly variable, especially during cloudy days or under a canopy, and can interfere with other environmental factors such as temperature and water availability.
• With the aim of decoupling the effect of radiation and temperature from water availability, we conducted an experiment where two perennial and three summer semi‐deciduous shrub species were subjected to different levels of irradiation. In order to follow plant responses to light exposure, we measured gas exchange, photosystem II photochemical efficiency, photosynthetic pigments and leaf mass area in spring and summer.
• Results showed that all study species presented a plastic response to different light conditions, and that light‐related traits varied in a coordinated manner. Summer semi‐deciduous species exhibited a more opportunistic response, with higher photosynthesis rates in full sun, but under shade conditions, the two strategies presented similar assimilation rates. Stomatal conductance did not show such a drastic response as photosynthetsis, being related to changes in WUE. Daily cycles of F_v/F_m revealed a slight photoinhibitory response during summer, mainly in perennial species. In all cases photosynthetic pigments adjusted to the radiation level; leaves had lower chlorophyll content, higher pool of xanthophylls and higher proportion of the de‐epoxydaded state of xanthophylls under sun conditions. Lutein content increased in relation to the xanthophyll pool under shade conditions.
• Our results evidenced that radiation is an important driving factor controlling morphological and physiological status of Mediterranean shrub species, independently of water availability. Summer semi‐deciduous species exhibit a set of traits with higher response variability, maximising their photosynthetic assimilation under different sun conditions.

     

Effects of salt stress on low molecular antioxidants and redox state of plastoquinone and P700 in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> (glycophyte) and <Emphasis Type="Italic">Eutrema salsugineum</Emphasis> (halophyte)

The effects of NaCl treatment were analysed in two species of considerably different resistance. In glycophyte, the content of ascorbate decreased but lipophilic antioxidants (α-tocopherol, plastochromanol, and hydroxy-plastochromanol) increased due to 150 mM NaCl. In halophyte, 300 mM NaCl caused a significant increase in hydrophilic antioxidants (ascorbate, total glutathione) but not in the lipophilic antioxidants. The redox states of plastoquinone (PQ) and P700 were also differently modulated by salinity in both species, as illustrated by an increased oxidation of these components in glycophyte. The presented data suggest that E. salsugineum was able to avoid a harmful singlet oxygen production at PSII, which might be, at least in part, attributed to the induction of the ascorbate-glutathione cycle. Another important cue of a high salinity resistance of this species might be the ability to sustain a highly reduced states of PQ pool and P700 under stress, which however, drastically affect the NADPH yield.     

Switch from inhibition to activation of the mitochondrial permeability transition during hematoporphyrin-mediated photooxidative stress.: Unmasking pore-regulating external thiols

We have studied the mitochondrial permeability transition pore (PTP) under oxidizing conditions with mitochondria-bound hematoporphyrin, which generates reactive oxygen species (mainly singlet oxygen, ¹O₂) upon UV/visible light-irradiation and promotes the photooxidative modification of vicinal targets. We have characterized the PTP-modulating properties of two major critical sites endowed with different degrees of photosensitivity: (i) the most photovulnerable site comprises critical histidines, whose photomodification by vicinal hematoporphyrin causes a drop in reactivity of matrix-exposed (internal), PTP-regulating cysteines thus stabilizing the pore in a closed conformation; (ii) the most photoresistant site coincides with the binding domains of (external) cysteines sensitive to membrane-impermeant reagents, which are easily unmasked when oxidation of internal cysteines is prevented. Photooxidation of external cysteines promoted by vicinal hematoporphyrin reactivates the PTP after the block caused by histidine photodegradation. Thus, hematoporphyrin-mediated photooxidative stress can either inhibit or activate the mitochondrial permeability transition depending on the site of hematoporphyrin localization and on the nature of the substrate; and selective photomodification of different hematoporphyrin-containing pore domains can be achieved by fine regulation of the sensitizer/light doses. These findings shed new light on PTP modulation by oxidative stress.     

fur (-) mutation increases the survival time of Escherichia coli under photooxidative stress in aquatic environments

We investigated the survival of the wild type Escherichia coli (QC771) and fur- mutant strain (QC1732) under photooxidative stress in different water sources. The survival of fur- mutant and wild type E. coli was seen as a significant decrease in the visible light samples in the presence of methylene blue (MB). The fur-E. coli strain lived longer than the wild type E. coli strain on exposure to MB and visible light, which generates singlet oxygen, in both lake water (48-h) and pure water (16-h). It is interesting to note that the survival of both wild type and the fur- mutant strain was more protected at 24 °C than at other temperatures. The Fur protein does not have any relation to the entry of E. coli into the viable but nonculturable state (VBNC) under photooxidative stress. This is the first study which shows that fur- mutation increases the resistance of E. coli to photooxidative stress in aquatic environments, and the Fur protein does not have any relation to the entry of E. coli into the VBNC state.     

Plastoquinol is more active than α-tocopherol in singlet oxygen scavenging during high light stress of Chlamydomonas reinhardtii

In the present study, we have performed comparative analysis of different prenyllipids in Chlamydomonas reinhardtii cultures during high light stress under variety of conditions (presence of inhibitors, an uncoupler, heavy water). The obtained results indicate that plastoquinol is more active than α-tocopherol in scavenging of singlet oxygen generated in photosystem II. Besides plastoquinol, also its oxidized form, plastoquinone shows antioxidant action during the stress conditions, resulting in formation of plastoquinone-C, whose level can be regarded as an indicator of singlet oxygen oxidative stress in vivo. The pronounced stimulation of α-tocopherol consumption and α-tocopherolquinone formation by an uncoupler, FCCP, together with the results of additional model system studies, led to the suggestion that α-tocopherol can be recycled in thylakoid membranes under high light conditions from 8a-hydroperoxy-α-tocopherone, the primary oxidation product of α-tocopherol by singlet oxygen.     

Photoinhibition of Photosynthesis: Role of Carotenoids in Photoprotection of Chloroplast Constituents

Exposure of plants to irradiation, in excess to saturate photosynthesis, leads to reduction in photosynthetic capacity without any change in bulk pigment content. This effect is known as photoinhibition. Photoinhibition is followed by destruction of carotenoids (Cars), bleaching of chlorophylls (Chls), and increased lipid peroxidation due to formation of reactive oxygen species if the excess irradiance exposure continues. Photoinhibition of photosystem 2 (PS2) in vivo is often a photoprotective strategy rather than a damaging process. For sustainable maintenance of chloroplast function under high irradiance, the plants develop various photoprotective strategies. Cars perform essential photoprotective roles in chloroplasts by quenching the triplet Chl and scavenging singlet oxygen and other reactive oxygen species. Recently photoprotective role of xanthophylls (zeaxanthin) for dissipation of excess excitation energy under irradiance stress has been emphasised. The inter-conversion of violaxanthin (Vx) into zeaxanthin (Zx) in the light-harvesting complexes (LHC) serves to regulate photon harvesting and subsequent energy dissipation. De-epoxidation of Vx to Zx leads to changes in structure and properties of these xanthophylls which brings about significant structural changes in the LHC complex. This ultimately results in (1) direct quenching of Chl fluorescence by singlet-singlet energy transfer from Chl to Zx, (2) trans-thylakoid membrane mediated, pH-dependent indirect quenching of Chl fluorescence. Apart from these, other processes such as early light-inducible proteins, D1 turnover, and several enzymatic defence mechanisms, operate in the chloroplasts, either for tolerance or to neutralise the harmful effect of high irradiance.