硫磺菌原变种液体培养代谢物生物活性分析

硫磺菌原变种Laetiporus sulphureusvar.sulphureus在液体培养条件下对果蝇具有致死效应,研究发现在液体培养过程中分泌到细胞外的代谢产物是致死效应的主要原因,并且上清液对果蝇的生物活性受pH值的影响。离子交换树脂柱和高效液相色谱分离分析表明草酸存在于硫磺菌原变种培养液的上清液中并且是果蝇致死效应和培养体系pH下降的一个重要因素。硫磺菌原变种在气升式反应器ALR/ff培养体系中草酸的浓度、菌丝体量和pH值呈简单相关。进一步分析发现还有另外一种结构未知、在碱性条件下呈紫红色的色素也具有致死效应。     

Protective Role of Thidiazuron Treatment on Cucumber Seedlings Exposed to Heavy Metals and Chilling

The action of thidiazuron, a synthetic growth regulator, was studied on 7-day-old cucumber seedlings (Cucumis sativus L., cv. Monastyrskii) exposed to chilling and sublethal concentrations of lead and copper ions. The extent of injury was assessed from the electrolyte leakage from cotyledonary leaves into distilled water. Separate application of each stress factor induced an increase in membrane permeability; however, their combined application caused a weaker response. A preliminary treatment of seedlings with thidiazuron fully or partly prevented the stress-induced stimulation of electrolyte leakage from cotyledon segments. It is concluded that thidiazuron elevates plant resistance to adverse environments.     

The Effects of Root Treatment with Various Stress Agents on Plant Cold- and Heat-Tolerance

The root systems of cucumber (Cucumis sativus L.), wheat (Triticum aestivum L.), and barley (Hordeum vulgare L.) plants were subjected to a short-term (6–7-h-long) treatment with low and high temperatures, sodium chloride, and lead nitrate, and the effect of these treatments on the changes in the cold- and heat-tolerance of leaf cells was determined. It was established that chilling of cucumber and wheat seedling roots at 10 and 2°C, respectively, or their heating at 38°C and 40°C, respectively, induced an increase in both cold- and heat-tolerance of leaf cells. An increase in the cold- and heat-tolerance was also observed in roots treated with sodium chloride at concentrations of 0.15 M (cucumber) and 0.2 M (wheat), as well as with lead nitrate at a concentration of 0.1 mM (barley, wheat). The tolerance increase induced by these stress agents was accompanied by a considerable increase in the ABA concentration in leaves. The effect of physical and chemical stress agents is suggested to induce the same nonspecific changes in the aboveground organs. These changes bring about, directly or indirectly, an increase in the cold- and heat-tolerance and are related to an increase in the ABA content.     

Effect of the Pool Size of Stromal Reductants on the Alternative Pathway of Electron Transfer to Photosystem I in Chloroplasts of Intact Leaves

The effect of elevated temperature on electron flow to plastoquinone pool and to PSI from sources alternative to PSII was studied in barley (Hordeum vulgare L.) and maize (Zea mays L.) leaves. Alternative electron flow was characterized by measuring variable fluorescence of chlorophyll and absorption changes at 830 nm that reflect redox changes of P700, the primary electron donor of PSI. The treatment of leaves with elevated temperature resulted in a transient increase in variable fluorescence after cessation of actinic light. This increase was absent in leaves treated with methyl viologen (MV). The kinetics of P700⁺ reduction in barley and maize leaves treated with DCMU and MV exhibited two exponential components. The rate of both components markedly increased with temperature of the heat pretreatment of leaves when the reduction of P700⁺ was measured after short (1 s) illumination of leaves. The acceleration of both kinetic components of P700⁺ reduction by high-temperature treatment was much less pronounced when P700⁺ reduction rate was measured after illumination of leaves for 1 min. Since the treatment of leaves with DCMU and MV inhibited both the electron flow to PSI from PSII and ferredoxin-dependent cycling of electrons around PSI, the accelerated reduction of P700⁺ indicated that high temperature treatment activated electron flow to PSII from reductants localized in the chloroplast stroma. We conclude that the lesser extent of activation of this process by elevated temperature after prolonged illumination of heat-inhibited leaves is caused by depletion of the pool stromal reductants in light due to photoinduced electron transfer from these reductants to oxygen.     

The ontogenetic shift in thermoregulatory behaviour of newt larvae: testing the 'enemy-free temperatures' hypothesis

Growth and development affect life-history traits, and consequently organismal fitness. The inevitable increase in body size during ontogeny is associated with changes in both resource use and predation risk, which results in the ontogenetic shift in habitat preferences. In this study, we examined whether the shift in preferred body temperatures ( T _ps) of newt larvae Triturus alpestris increases the T _ps deviation of the most vulnerable stages (after hatching and during metamorphosis) from the T _ps range of their main predator, dragonfly larvae Aeshna cyanea – the 'predator-free temperatures' hypothesis. Analyses of thermoregulatory behaviour in the laboratory thermal gradient showed that freshly hatched newt larvae maintained lower water temperatures than larvae in later stages, whose T _ps largely fell into the T _ps range of dragonfly larvae. With respect to the thermal quality of natural habitat, the anti-predator effectiveness of the T _ps shift decreased during development. Water temperatures in natural habitats were located largely below the preferred body temperature range of both newt and dragonfly larvae, which limits their potential thermal niche partitioning. We conclude that factors other than predator avoidance drive the ontogenetic T _ps shift in our model system.     

Hypoxia inhibits maturation and trafficking of hERG K channel protein: Role of Hsp90 and ROS

We previously reported that reactive oxygen species (ROS) generated during hypoxia decrease hERG current density and protein expression in HEK cells stably expressing hERG protein. In the present study, we investigated the molecular mechanisms involved in hypoxia-induced downregulation of hERG protein. Culturing cells at low temperatures and addition of chemical chaperones during hypoxia restored hERG expression and currents to normoxic levels while antiarrhythmic drugs, which selectively block hERG channels, had no effect on hERG protein levels. Pulse chase studies showed that hypoxia blocks maturation of the core glycosylated form in the endoplasmic reticulum (ER) to the fully glycosylated form on the cell surface. Co-immunoprecipitation experiments revealed that hypoxia inhibited interaction of hERG with Hsp90 chaperone required for maturation, which was restored in the presence of ROS scavengers. These results demonstrate that ROS generated during hypoxia prevents maturation of the hERG protein by inhibiting Hsp90 interaction resulting in decreased protein expression and currents.     

Free amino acid composition in blood and muscle of the gobi <Emphasis Type="Italic">Precottus glehni</Emphasis> at the period of preparation and completion of hibernation

The freshwater fish gobi Preccottus glehni survives after wintering in ponds frozen in winter till the very bottom. In adaptation of poikilothermal animals to wintering at near-zero temperatures, an essential role is played by free amino acids; accumulation of a large amount of some particular acid at the period of preparation to the state of hibernation can indicate the protective role of this acid in the low-temperature adaptation. In the present work it has been shown that as soon as by the end of August, in the gobi muscle, the taurine concentration increases three times as compared with that in July, the taurine pool after this reaching 50% of the total pool of free amino acids in the muscle tissue. During December and after the 3-month hibernation in ice, it exceeds the April and July levels 8 and 4 times, respectively, and amounts to 50% of the total free amino acid pool for muscle and to 40% for blood. Several days prior to the beginning of winter hibernation under natural conditions, both in blood and in muscle, there is revealed disappearance or a sharp fall of concentration of essential amino acids. An essential peculiarity of the change in the free amino acid composition after hibernation was a significant rise of alanine concentration in muscle—3.5 times as compared with July and 1.4–1.8 times as compared with changes in December. The total free amino acid pool in muscle in December as compared with that in July increased almost 1.5 times (34.76 ± 1.12 μmol/g wet weight), while after hibernation—2 times. Peculiarities of taurine accumulation long before the beginning of winter cold allow suggesting that role of taurine consists in providing a possibility of existence of eurythermal fish at near-zero temperature.     

A graphical method for identifying the six types of non‐deep physiological dormancy in seeds

We present a new seed dormancy classification scheme for the non‐deep level of the class physiological dormancy (PD), which contains six types. Non‐deep PD is divided into two sublevels: one for seeds that exhibit a dormancy continuum (types 1, 2 and 3) and the other for those that do not exhibit a dormancy continuum (types 4, 5 and 6). Analysis of previous studies showed that different types of non‐deep PD also can be identified using a graphical method. Seeds with a dormancy (D) ? conditional dormancy (CD) ? non‐dormancy (ND) cycle have a low germination percentage in the early stages of CD, and during dormancy loss the germination capacity increases. However, seeds with a CD/ND (i.e. D→CD?ND) cycle germinate to a high percentage at a narrow range of temperatures in the early stages of CD. Cardinal temperatures for seeds with either a D/ND or a CD/ND cycle change during dormancy loss: the ceiling temperature increases in seeds with Type 1, the base temperature decreases in seeds with Type 2 and the base and ceiling temperatures decrease and increase, respectively, in seeds with Type 3. Criteria for distinguishing the six types of non‐deep PD and models of the temperature functions of seeds with types 1, 2 and 3 with both types of dormancy cycles are presented. The relevancy of our results to modelling the timing of weed seedling emergence is briefly discussed.     

Tryptophan scanning mutagenesis of the first transmembrane domain of the innexin Shaking-B(Lethal)

The channel proteins of gap junctions are encoded by two distinct gene families, connexins, which are exclusive to chordates, and innexins/pannexins, which are found throughout the animal kingdom. Although the relationship between the primary structure and function of the vertebrate connexins has been relatively well studied, there are, to our knowledge, no structure-function analyses of invertebrate innexins. In the first such study, we have used tryptophan scanning to probe the first transmembrane domain (M1) of the Drosophila innexin Shaking-B(Lethal), which is a component of rectifying electrical synapses in the Giant Fiber escape neural circuit. Tryptophan was substituted sequentially for 16 amino acids within M1 of Shaking-B(Lethal). Tryptophan insertion at every fourth residue (H27, T31, L35, and S39) disrupted gap junction function. The distribution of these sites is consistent with helical secondary structure and identifies the face of M1 involved in helix-helix interactions. Tryptophan substitution at several sites in M1 altered channel properties in a variety of ways. Changes in sensitivity to transjunctional voltage (Vj) were common and one mutation (S39W) induced sensitivity to transmembrane voltage (Vm). In addition, several mutations induced hemichannel activity. These changes are similar to those observed after substitutions within the transmembrane domains of connexins.     

Toxicity of Crude Extracellular Products of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis> on Rohu, <Emphasis Type="Italic">Labeo rohita</Emphasis> (Ham.)

In the present study the haemolytic and proteolytic activity of extracellular products (ECP) secreted from Aeromonas hydrophila (CAHH14 strain) were studied with respect to temperature and different time of incubation as well as its lethal toxicity on rohu, Labeo rohita. The strain was isolated from Catla catla (showing abdominal dropsy symptom) collected from the pond of Central Institute of Freshwater Aquaculture (CIFA), Bhubaneswar, India and was characterized on the basis of biochemical tests. The highest production of haemolysin was achieved when the bacteria was grown at 35°C for 30 h. The proteolytic activity was found to be highest when the bacterium was grown at 30°C for 36 h. The haemolytic and proteolytic toxin produced by Aeromonas hydrophila was found to be lethal to rohu (LD₅₀ 1.7 × 10⁴ cfu/ml). The lethality of ECP was decreased by heating and completely inactivated by boiling at 100°C for 10 min. This indicates that protease activity and haemolytic activity of A. hydrophila ECP was temperature dependant.