Draft genome sequence of Pseudomonas fuscovaginae, a broad-host-range pathogen of plants

Pseudomonas fuscovaginae was first reported as a pathogen of rice causing sheath rot in plants grown at high altitudes. P. fuscovaginae is now considered a broad-host-range plant pathogen causing disease in several economically important plants. We report what is, to our knowledge, the first draft genome sequence of a P. fuscovaginae strain.     

Thermal ecology of the mudskippers, Periophthalmus koelreuteri (Pallas) and Boleophthalmus boddarti (Pallas) of Kuwait Bay

The annual range of body temperatures (14–35°C) of emergent mudskippers are substantially less than that of air temperatures (10–42°C) as a result of behavioural thermoregulation. In winter, low surface temperatures are avoided by remaining in burrows. Newly emerged mudskippers then bask until body temperatures rise above 14°C before they move onto the mud. In summer, body temperatures are kept lower than ambient by selecting areas where evaporative cooling is high. Body temperatures generally match those of wet mud, which can be 7°C lower than air shade temperatures. The smaller, more terrestrial, Periophthalmus koelreuteri have body temperatures which are mainly lower in summer and higher in winter than Boleophthalmus boddarti .     

Thermal ecology and spatio-temporal distribution of the Mediterranean lizard Psammodromus algirus

We studied summer activity patterns and thermoregulation in the Mediterranean lizard Psammodromus algirus in a holm oak wood in Central Spain. The circadian rhythm curve was bimodal, with a pronounced peak after sunrise, a minimum at noon and a second lower peak in the afternoon. Increased activity in the morning could be explained by insolation levels, whereas the midday low was caused by the rise in temperature.
There was a strong positive correlation between body temperature and both air and ground temperatures. Nevertheless, there was no significant correlation between body and air temperatures when the effect of radiation was removed, which was done by considering only individuals in the shade at high temperatures.
Activity showed a bell-shaped relationship with body and air temperatures, with maximum activity at 24.5°C (air temperature) and 31.4°C (body temperature). The spatial distribution pattern of P. algirus (selection of sunny areas and distance to the vegetation edge) reflects the environmental temperatures that are optimal for activity; the lizards show a strong preference for sunny patches when air temperature is lower than the optimum. We conclude that heliothermy is the only viable thermoregulation pattern in Mediterranean forest environments, with heat conduction playing a negligible rote.     

Revision of the relationship between anther morphology and pollen sterility by cold stress at the booting stage in rice

Background and AimsCold stress in rice (Oryza sativa) plants at the reproductive stage prevents normal anther development and causes pollen sterility. Tapetum hypertrophy in anthers has been associated with pollen sterility in response to cold at the booting stage. Here, we re-examined whether the relationships between anther abnormality and pollen sterility caused by cold stress at the booting stage in rice can be explained by a monovalent factor such as tapetum hypertrophy.MethodsAfter exposing plants to a 4-d cold treatment at the booting stage, we collected and processed anthers for transverse sectioning immediately and at the flowering stage. We anatomically evaluated the effect of cold treatment on anther internal morphologies, pollen fertilities and pollen numbers in the 13 cultivars with various cold sensitivities.Key ResultsWe observed four types of morphological anther abnormalities at each stage. Pollen sterility was positively correlated with the frequency of undeveloped locules, but not with tapetum hypertrophy as commonly believed. In cold-sensitive cultivars grown at low temperatures, pollen sterility was more frequent than anther morphological abnormalities, and some lines showed remarkably high pollen sterility without any anther morphological alterations. Most morphological anomalies occurred only in specific areas within large and small locules. Anther length tended to shorten in response to cold treatment and was positively correlated with pollen number. One cultivar showed a considerably reduced pollen number, but fertile pollen grains under cold stress. We propose three possible relationships to explain anther structure and pollen sterility and reduction due to cold stress.ConclusionsThe pollen sterility caused by cold stress at the booting stage was correlated with the frequency of entire locule-related abnormalities, which might represent a phenotypic consequence, but not a direct cause of pollen abortion. Multivalent factors might underlie the complicated relationships between anther abnormality and pollen sterility in rice.     

Isolation of insect pathogenic bacteria, Providencia rettgeri, from Heterorhabditis spp.

Nematodes of the genus Heterorhabditis carry bacteria of the genus Photorhabdus into insects including pests of horticultural crops. The bacteria kill the insect and provide conditions which allow for the growth and development of the nematodes. It is reported here that the majority of Heterorhabditis spp. strains tested contained a second bacterial species which was identified as Providencia rettgeri. Injection of the bacteria into waxmoth larvae showed that P. rettgeri was at least as pathogenic as Photorhabdus sp. K122. Both had LD₅₀ values of less than one bacterial cell/larva, but P. rettgeri killed the insects at a considerably faster rate than K122 at both 28°C and 9°C. Since Photorhabdus kills very slowly at low temperatures, it appeared that P. rettgeri might be a better pest control agent under these conditions. However, P. rettgeri was not pathogenic when carried into insect larvae by the nematode, indicating that the nematode suppressed either its release or pathogenicity. It will be necessary to find ways of bypassing or inhibiting this suppression for P. rettgeri to fulfil its potential in pest control.     

Seed germination ecology of Portulaca oleracea L.: an important weed of rice and upland crops

Portulaca oleracea , a C₄ species, is reported to be a serious weed in 45 crops in 81 countries. Experiments were conducted in the laboratory, the screenhouse and the field to determine the influence of environmental factors on seed germination and seedling emergence of P. oleracea . In the laboratory, germination in the dark was low and was not influenced by the tested temperatures (35/25°C, 30/20°C and 25/15°C alternating day/night temperatures). In the light/dark regime, however, germination was lower at 25/15°C and 35/25°C than at 30/20°C (70%, 75% and 81% germination, respectively). In conditions of 106 mM sodium chloride or −0.34 MPa osmotic potential, seeds germinated to only 50% of maximum germination of the control. Germination was not influenced by buffered pH solutions ranging from 5 to 9. In the screenhouse, germination was greatest for seeds placed on the soil surface, but emergence declined with increasing seed burial depth in soil; no seedlings emerged from the depth of 2 cm. Seedling emergence and seedling dry matter were markedly reduced by the addition of rice residue to the soil surface at rates equivalent to 4 to 6 t ha⁻¹. In the field, seedling emergence of P. oleracea was greater under zero till (ZT) (17–20%) than under minimum tillage (6–10%), a likely reflection of low seed burial and exposure of seeds to light with a ZT system. This study identifies some of the factors enabling P. oleracea to be a widespread weed in the humid tropics, and the information could contribute to improved control strategies.     

Thermophilic methanogens in rice field soil

The soil temperature in flooded Italian rice fields is generally lower than 30°C. However, two temperature optima at ≈ 41°C and 50°C were found when soil slurries were anoxically incubated at a temperature range of 10–80°C. The second temperature optimum indicates the presence of thermophilic methanogens in the rice field soil. Experiments with ¹⁴C-labelled bicarbonate showed that the thermophilic CH₄ was exclusively produced from H₂/CO₂. Terminal restriction fragment length polymorphism (T-RFLP) of archaeal SSU rRNA gene fragments revealed a dramatic change in the archaeal community structure at temperatures > 37°C, with the euryarchaeotal rice cluster I becoming the dominant group (about 80%). A clone library of archaeal SSU rRNA gene fragments generated at 49°C was also dominated (10 out of 11 clones) by rice cluster I. Our results demonstrate that Italian rice field soil contains thermophilic methanogenic activity that was most probably a result of members of the as yet uncultivated euryarchaeotal rice cluster I.     

Temperature influences walking speed and walking activity of Trichogramma brassicae (Hym., Trichogrammatidae)

Walking speeds and walking activities (walking time divided by total time) of Trichogramma brassicae were determined at 12, 16, 20 and 25°C. Walking speed was measured during a 5-min period, and walking activity over a 4-day period. Both walking speed and walking activity were strongly influenced by temperature. Walking speed increased linearly with the temperature and was twice as high at 20 as at 12°C. At 25°C, walking activity was high during the whole day, at 20 and 16°C it decreased during the afternoon, whereas at 12°C the wasps became most active only at noon or later. At low temperatures, there was a strong individual variation in walking activity. At 25°C, T. brassicae was active most of the time, so the area searched per time unit can only increase at temperatures above 25°C if walking speed increases. At temperatures below 20°C, searching was more restricted by low walking activity than by low walking speed. Even disregarding other effects of temperature, the reduction in walking speed and walking activity at suboptimal temperatures means that T. brassicae can only search half of the area at 20°C, and only one-seventh at 15°C that it can search at 25°C.     

Survival at low temperature of larvae of the pine processionary moth Thaumetopoea pityocampa from an area of range expansion

1 Larvae of Thaumetopoea pityocampa (Lepidoptera: Notodontidae) develop throughout the winter, although their feeding activity and survival can be impaired by adverse climatic factors. The present study investigated the survival at low temperature of larvae originating from a population with range expansion in an alpine valley in Northern Italy.
2 The supercooling point of individually analysed larvae averaged at −7 °C. This value insufficiently described the cold hardiness of the larvae; 39% of the tested larvae were alive when returned to room temperature immediately after freezing. When larval colonies inside their nest were exposed to −17 °C for 1 h after gradual temperature decrease, survival was 70.4%.
3 Rearing of larvae in the laboratory at different day/night temperatures indicated an effect of cumulative chill injury on larvae. A logistic regression explained the relationship between negative thermal sum (h°C below 0 °C) received in the laboratory experiment and larval survival. A similar relationship was demonstrated between negative thermal sum and survival of larval colonies in the field.
4 In the laboratory experiment, some tested larvae were able to survive for up to 8 weeks without feeding depending on rearing temperature. As expected, feeding occurred only when larvae were reared at temperatures of 9 °C day/0 °C night.
5 We classify the larvae of T. pityocampa as being moderate freezing tolerant. The winter behaviour allows this species to track climate warming by a rapid expansion into those areas that become compatible with the insect's development.     

Thermophilic sulfate-reducing bacteria in cold marine sediment

Abstract Sulfate reduction was measured with the ³⁵SO₄²⁻ -tracer technique in slurries of sediment from Aarhus Bay, Denmark, where seasonal temperatures range from 0° to 15°C. The incubations were made at temperatures from 0°C to 80°C in temperature increments of 2°C to search for presence of psychrophilic, mesophilic and thermophilic sulfate-reducing bacteria. Detectable activity was initially only in the mesophilic range, but after a lag phase sulfate reduction by thermophilic sulfate-reducing bacteria were observed. No distinct activity of psychrophilic sulfate-reducing bacteria was detected. Time course experiments showed constant sulfate reduction rates at 4°C and 30°C, whereas the activity at 60°C increased exponentially after a lag period of one day. Thermophilic, endospore-forming sulfate-reducing bacteria, designated strain P60, were isolated and characterized as D esulfotomaculum kuznetsovii . The temperature response of growth and respiration of strain P60 agreed well with the measured sulfate reduction at 50°–70°C. Bacteria similar to strain P60 could thus be responsible for the measured thermophilic activity. The viable population of thermophilic sulfate-reducing bacteria and the density of their spores was determined in most probable number (MPN) dilutions. The density was 2.8·10⁴ cells·.g⁻¹ fresh sediment, and the enumerations suggested that they were all present as spores. This result agrees well with the observed lag period in sulfate reduction above 50°C. No environment with temperatures supporting the growth of these thermophiles is known in the region around Aarhus Bay.     

Temperature modulates the cell wall mechanical properties of rice coleoptiles by altering the molecular mass of hemicellulosic polysaccharides

The present study was conducted to investigate the mechanism inducing the difference in the cell wall extensibility of rice ( Oryza sativa L. cv. Koshihikari) coleoptiles grown under various temperature (10–50°C) conditions. The growth rate and the cell wall extensibility of rice coleoptiles exhibited the maximum value at 30–40°C, and became smaller as the growth temperature rose or dropped from this temperature range. The amounts of cell wall polysaccharides per unit length of coleoptile increased in coleoptiles grown at 40°C, but not at other temperature conditions. On the other hand, the molecular size of hemicellulosic polysaccharides was small at temperatures where the cell wall extensibility was high (30–40°C). The autolytic activities of cell walls obtained from coleoptiles grown at 30 and 40°C were substantially higher than those grown at 10, 20 and 50°C. Furthermore, the activities of (1→3),(1→4)- β -glucanases extracted from coleoptile cell walls showed a similar tendency. When oat (1→3),(1→4)- β -glucans with high molecular mass were incubated with the cell wall enzyme preparations from coleoptiles grown at various temperature conditions, the extensive molecular mass downshifts were brought about only by the cell wall enzymes obtained from coleoptiles grown at 30–40°C. There were close correlations between the cell wall extensibility and the molecular mass of hemicellulosic polysaccharides or the activity of β -glucanases. These results suggest that the environmental temperature regulates the cell wall extensibility of rice coleoptiles by modifying mainly the molecular mass of hemicellulosic polysaccharides. Modulation of the activity of β -glucanases under various temperature conditions may be involved in the alteration of the molecular size of hemicellulosic polysaccharides.     

Effects of constant and fluctuating temperatures on life span of Aedes taeniorhynchus adults

Male and virgin female Aedes taeniorhynchus were maintained on a sugar solution at constant temperatures, at split-temperatures, and at alternating temperatures from immediately after emergence until death in order to study the effect of temperatures on their longevity. Life spans were found to be temperature dependent at constant temperatures of 22, 27, and 32°C, but they were divided into ‘ageing’ and ‘dying’ phases at split-temperatures. The rate of ageing, which was independent of temperatures, was the same in males whether they were transferred from low to high or high to low temperatures. The rate of ageing was the same in females transferred from 22°C or 27 to 32°C, but much longer than expected when transferred from 22 to 27°C. Also, the rate of ageing was the same for females transferred from 32°C to either 22 or 27°C and from 27 to 22°C. The rate of dying was essentially temperature dependent in both males and females with slight temperature compensation occurring in some cases. Life spans were the same in males when alternated between 22?27°C, 27?32°C, and 22?32°C. In females they were same at 27?32°C and 22?32°C, but were much longer than expected at 22?27°C. It is concluded that the threshold theory is confirmed when mosquitoes are maintained at split-temperatures and at alternating temperatures in their optimum range of temperatures.     

Wheat catalase expressed in transgenic rice can improve tolerance against low temperature stress

We examined whether the expression of wheat catalase (EC 1.11.1.6) cDNA in transgenic rice ( Oryza sativa L.) could enhance tolerance against low temperature injury. Transgenic rice plants expressing wheat CAT protein showed an increase of activities in leaves at 25°C, 2- to 5-fold that in non-transgenic rice. At 5°C, catalase activities were about 4–15 times higher than those in non-transgenic rice were. A comparison of damage observed in leaves as they withered due to chilling at 5°C showed that transgenic rice displayed an increased capability to resist low temperature stress. The exposure of these plants to low temperature at 5°C for 8 days resulted in decreased catalase activities in leaves at 25°C, but the transgenic plants indicated 4 times higher residual catalase activities than those of non-transgenic ones. The concentration of H₂O₂ in leaves was kept lower in transgenic rice than that of the control plants during the 8 days chilling. These results suggest that the improved tolerance against low temperature stress in genetically engineered rice plants be attributed to the effective detoxification of H₂O₂ by the enhanced catalase activities.     

Adaptation to sub- and supraoptimal temperatures of inbred maize lines differing in origin with regard to seedling development and photosynthetic traits

Six inbred lines of maize ( Zea mays L.) from cool temperate regions (C) and from warm regions (W) were grown at 14, 22, 30 and 38°C up to the same physiological age, the full expansion of the third leaf. Generally, plants developed smaller shoot dry weights and leaf areas at extreme temperatures. The shoot:root ratio was lowest at 22°C and highest at 30°C. Most lines had a minimum for specific leaf dry weight at 30°C, but W lines had a second lower minimum at 14°C. Phosphofructokinase activity scarcely reacted to temperature between 22° and 38°C; at 14°C one C line and all W lines had rather low activities. Generally, the chlorophyll content increased steeply from 14 to 22°C and decreased somewhat from 30 to 38°C. In C lines the carotenoid level decreased from 14 to 38°C. No uniform temperature response was found for PEP carboxylase activity, but the highest activity was mostly attained at 38°C. RuBP carboxylase activity increased considerably from 14 to 22°C and remained comparatively constant at higher temperatures. The highest activity of NADP malate dehydrogenase was found at 22°C, with a decrease up to 38°C and with second lowest values at 14°C. C lines possessed larger leaf areas, shoot dry weights and higher shoot:root ratios than W lines at 14 and 22°C, and higher specific leaf dry weights over the whole temperature range. The genotypic pattern of shoot dry weight at 14°C corresponded reasonably well with that of phosphofructokinase activity. A better adaptation of C lines to suboptimal temperatures was mostly clearly indicated for photosynthetic traits which have a well proven relationship with the chloroplast membranes: chlorophyll, carotenoids and RuBP carboxylase. The least distinct effects of origin were observed at 38°C; a tendency prevailed for a better performance of C lines with regard to phosphofructokinase, carotenoids, RuBP carboxylase and NADP malate dehydrogenase.     

Thermal preference in the pool frog Rana lessonae: impact on the reproductive behaviour of a northern fringe population

A documented warmth preference in the pool frog Rana lessonae is also expressed in the reproductive behaviour of a northern fringe population (60°N). During the breeding period of 31 days, individually marked frogs were observed during chorus. Male choruses were formed in the warmest parts of the pond, but moved between three areas. At night, these areas maintained the highest water temperatures in the pond, especially at low temperatures prevailing in the beginning of the breeding period. Oviposition occurred in the current chorus areas, and 86% took place during the first two weeks of the breeding period. That the choruses and oviposition changed locations within the favourable breeding areas can be explained by a pronounced thermal preference in the frogs, and winds moving the warm surface water in different directions.
The water temperature required to initiate oviposition was 16°C, which is similar to those reported from related more southern populations. The thermal preference expressed in the breeding behaviour of both sexes and the tendency for early oviposition may be behavioural responses to environmental constraints on reproduction in the short season at 6O°N.     

Generation of a membrane-bound, oligomerized pre-pore complex is necessary for pore formation by Clostridium septicum alpha toxin

Low-temperature inhibition of the cytolytic activity of alpha toxin has facilitated the identification of an important step in the cytolytic mechanism of this toxin. When alpha toxin-dependent haemolysis was measured on erythrocytes at various temperatures it was clear that at temperatures ≤15°C the haemolysis rate was significantly inhibited with little or no haemolysis occurring at 4°C. Alpha toxin appeared to bind to and oligomerize on erythrocyte membranes with similar kinetics at 4°C and 37°C. The slight differences in these two processes at 4°C and 37°C could not account for the loss of cytolytic activity at low temperature. At 4°C alpha toxin neither stimulated potassium release from erythrocytes nor formed pores in planar membranes. In contrast, at temperatures ≥25°C both processes proceeded rapidly. Pores that were opened in osmotically stabilized erythrocytes could not be closed by low temperature. Therefore, low temperature appeared to prevent the oligomerized complex from forming a pore in the membrane. These data support the hypothesis that alpha toxin oligomerizes into a membrane-bound, pre-pore complex prior to formation of a pore in a lipid bilayer.     

Heat treatment to decrease chilling injury in tomato fruit. Effects on lipids, pericarp lesions and fungal growth

Tomato fruits are sensitive to low temperature and develop chilling injury, while at nonchilling temperatures they ripen rapidly. Previously, a hot-air treatment was found to reduce the sensitivity of the fruit to low temperatures. In the present study hot air was compared to hot water and their effects on reducing chilling injury and fungal decay were investigated. Tomatoes ( Lycopersicon esculentum cv. Daniella) at the breaker stage were subjected to hot air, 48 h at 38°C, or various hot water dips, 30 min at 40°C or 2 min at 46, 48 or 50°C, before holding at 2°C. The unheated tomatoes developed chilling injury and fungal infections at 2°C, but not at 12°C. All the heat treatments reduced chilling injury and decay in tomatoes held for 3 weeks at 2°C. The outer pericarp tissue of heated tomatoes had higher phospholipid and lower sterol contents than unheated tomatoes. Heated tomatoes also had less saturated fatty acids than unheated tomatoes held at 2°C, but not at 12°C. Scanning electron micrograph observations showed that all the fruits had microcracks in their surface, but the unheated chilled tomatoes had also fungal growth in the cracks, while those of the heated tomato fruit did not. In the areas of chilling injury collapsed cells were present under the peel and could also support pathogen development. It is suggested that the heat treatment institutes a response to high temperature stress in the fruit tissue that leads to strengthened membranes. This prevents the loss of function and cell collapse which was found in the chilling-injured areas of affected fruit.     

Effect of temperature on the vegetative growth of type and field strains of Bacillus cereus

Growth of eight selected potentially pathogenic strains of Bacillus cereus was evaluated in a rich medium at different temperatures. No strain grew at 50°C; maximal growth-permissive temperatures were in the range 46–50°C for six strains and under 46°C for one strain. Faster growth occurred at 42°C. Growth may be delayed at 20°C, where the lag phase can reach 7 h. Furthermore at 20°C, cells generally show an aggregation immediately in the early exponential stage, except for two strains. Owing to this aggregation, growth is more difficult to estimate by turbidimetry at lower temperatures. These data describe the behaviour of type and field strains between 50° and 20°C and can help the prediction of shelf-life of potentially contaminated products.     

Shoot temperatures and thermal tolerances for succulent species of Haworthia and Lithops

Abstract. Temperatures of small succulent plants, such as species in the genera Haworthia and Lithops , are highly influenced by temperatures of the surrounding soil. Indeed, the minimum and the maximum temperatures of the upper leaf epidermis of Haworthia retusa. H. turgida. Lithops leslei , and L. turbiniformis were generally within 1°C of the accompanying soil surface temperatures. An energybudget model closely predicted such soil-to-plant temperature differences as well as the effect of the greater convective exchange for the protruding Haworthia species compared with the Lithops species, which were flush with the soil surface. Although a lower shortwave absorptance would reduce maximum shoot temperatures, the shortwave absorptances of all four species were similar to those of the soil in their respective native habitats in South Africa. Tolerances of the four species to low and to high temperatures at three different day/night air temperatures (15°C/5°C, 30°C/20°C, and 45°C/35°C) were analysed using cellular accumulation of a vital stain, neutral red. Chlorenchyma cells were slightly more tolerant of extreme temperatures than were cells of the water-storage parenchyma. In this regard, H. retusa survived low and high temperatures that killed the water-storage parenchyma but not the chlorenchyma. Acclimation to low temperatures and to high temperatures, which was exhibited by all four species, led to estimated tolerances to 1 h at −16°C and 68°C. Although the low temperature tolerance is not particularly noteworthy, very few vascular plants are reportedly able to tolerate such high temperatures.     

Mode of high temperature injury to wheat. II. Comparisons of wheat and rice with and without inflorescences

High temperature injury to wheat ( Triticum aestivum L.) during grain development is manifested as acceleration of senescence. Experiments were conducted to elucidate the mode of senescence and site of high temperature responses. Wheat (cv. Chris) and rice ( Oryza sativa L. cv. Newbonnet), which have C₃ photosynthesis but different temperature responses, were grown with and without inflorescences under three temperature regimes after anthesis. Plant growth and constituents associated with senescence were measured weekly until physiological maturity. Increasing temperatures from 25°C/15°C to 35°C/25°C day/night after anthesis decreased growth, leaf viability, chlorophyll and protein concentrations, and RuBP carboxylase activity and increased protease and RNase activities in wheat. Inflorescence removal increased vegetative weights and slowed most senescence processes more in wheat than in rice, but did not alter the course of high temperature responses. Results are interpreted as indicating that diversion of nutrients from roots by inflorescence sinks at normal temperatures and by increased respiration at high temperatures caused similar responses. Source and sink activities appeared to be regulated jointly, probably by cytokinins from roots, during senescence at normal and elevated temperatures.