Does availability of potassium affect cold hardening of Scots pine through polyamine metabolism?

The effect of different potassium availability on the polyamines and frost resistance of Scots pine seedlings ( Pinus sylvestris L.) during cold hardening was studied. Scots pine seedlings were grown applying different rates of potassium by using the relative addition rate technique followed by a 2- or 9-week hardening period with decreased light intensity, day length and temperature. After 2 weeks of treatment the seedlings were not hardened (LT₅₀, =−11°C) and showed no differences in frost resistance, although differences in the polyamine levels between the K levels were observed. After 9 weeks of hardening the seedlings at the low, medium and high K levels showed a mean frost resistance (LTs.₅₀) of −81, −63, and −47°C, respectively. A negative effect of K on the frost resistance of the needles was also found in adult trees in September. The results indicate that at the early stage of cold hardening, potassium or free polyamine levels do not affect the frost resistance of Scots pine needles. However, in hardened seedlings and adult trees potassium displays a negative and putrescine a positive correlation with frost resistance, whereas spermidine and spermine show no correlation.     

Putrescine metabolism in excised cotyledons of Pinus radiata cultured in vitro

The metabolism of ¹⁴C-putrescine and the changes in the endogenous concentrations of putrescine, spermidine and spermine were studied when cotyledons of Pinus radiata D. Don were cultured under shoot-forming (SF, + N⁶-benzyladenine) and non-shoot-forming (NSF, - N⁶-benzyladenine) conditions. Differences in the total uptake of ¹⁴C-putrescine during a 2 h pulse feeding were not significant between the SF and NSF cotyledons except on day 3. The maximum uptake of label was on day 3 in the SF cotyledons, which released the highest amount of ¹⁴CO₂ as well. ¹⁴C from the labeled putrescine was incorporated mainly into γ-aminobutyric acid, aspartate and glutamate. High performance liquid chromatography of the endogenous polyamines indicated that spermidine was the most predominant polyamine in the cultured cotyledons of radiata pine. Spermine increased by about 60% in the SF and 25% in the NSF cotyledons between days 0 and 3 of culture.     

Requirement for far-red light to maintain secondary needle extension growth in northern but not southern populations of Pinus sylvestris (Scots pine)

Extension growth of secondary needles is under photoperiodic control in Pinus sylvestris . To test for the effects of far-red light on maintaining this extension growth, seedlings of six populations originating from latitudes between 57° and 67°N were raised for 11 weeks in continuous incandescent (metal halogen) light at 300 µmol m⁻² s⁻¹ and 20°C and then transferred at the same temperature to a daily regime of 8 h incandescent light (230 µmol m⁻² s⁻¹) followed by a 16 h day extension with cool white fluorescent light (40 µmol m⁻² s⁻¹, R/FR ratio 7.5) or with incandescent lamps (20 µmol m⁻² s⁻¹, R/FR ratio 2.0). For the seedlings from the three populations north of 64°, needle extension growth over 42 days in the FR-poor day extension treatment was lower by up to 40% than in the FR-rich day extension treatment, whereas for the seedlings from the three southern populations the needle extension growth was similar in both day extension treatments. The requirement for FR in day extensions is characteristic of 'light-dominant' photoperiodic control mechanisms. It appears that P. sylvestris changes from dark-dominant night timekeeping to light-dominant day timekeeping with increasing latitude, as with the photoperiodic control of budset in Picea abies .     

Involvement of polyamines in the interacting effects of low temperature and mineral supply on Pringlea antiscorbutica (Kerguelen cabbage) seedlings

Pringlea antiscorbutica, which is the sole endemic crucifer in the subantarctic zone, undergoes seedling development in a harsh and cold environment. Since, at the mature stage, this species exhibits several adaptations linked to cold tolerance such as high polyamine levels, potential adaptations and polyamine response were investigated in seedlings. In order to assess the specificity of responses, P. antiscorbutica was compared with Arabidopsis thaliana, which is characterized by a life cycle preventing cold exposure at seedling stage. P. antiscorbutica and A. thaliana seedlings were found to have strikingly contrasted responses to temperature changes and to mineral nutrition. Whereas A. thaliana seedlings showed the typical growth arrest of chilling-sensitive plants, P. antiscorbutica seedlings showed optimal root growth at low temperature (5/10 degrees C) and temperate conditions caused the early arrest of root growth. Cold tolerance was associated with increased levels of polyamines or with maintenance of high levels of polyamines. Comparison of both species showed that polyamine levels could be a significant marker of chilling tolerance in seedlings. Treatments with varying mineral supply showed a positive relationship between root growth rate and variations of agmatine and putrescine endogenous contents in roots of P. antiscorbutica. This may be the first demonstration that, even under conditions of accumulation induced by environmental stress, polyamine levels can still be correlated with developmental processes. Com parison of mineral supply and temperature effects strongly indicated a trade-off of polyamine involvement between development and response to stress.     

Thermal acclimation of photosynthesis in black spruce [Picea mariana (Mill.) B.S.P.

We investigated the thermal acclimation of photosynthesis and respiration in black spruce seedlings [ Picea mariana (Mill.) B.S.P.] grown at 22/14 °C [low temperature (LT)] or 30/22 °C [high temperature (HT)] day/night temperatures. Net CO₂ assimilation rates ( A _net) were greater in LT than in HT seedlings below 30 °C, but were greater in HT seedlings above 30 °C. Dark and day respiration rates were similar between treatments at the respective growth temperatures. When respiration was factored out of the photosynthesis response to temperature, the resulting gross CO₂ assimilation rates ( A _gross) was lower in HT than in LT seedlings below 30 °C, but was similar above 30 °C. The reduced A _gross of HT seedlings was associated with lower needle nitrogen content, lower ribulose 1·5-bisphosphate carboxylase/oxygenase (Rubisco) maximum carboxylation rates ( V _cmax) and lower maximum electron transport rates ( J _max). Growth treatment did not affect V _cmax :  J _max. Modelling of the CO₂ response of photosynthesis indicated that LT seedlings at 40 °C might have been limited by heat lability of Rubisco activase, but that in HT seedlings, Rubisco capacity was limiting. In sum, thermal acclimation of A _net was largely caused by reduced respiration and lower nitrogen investments in needles from HT seedlings. At 40 °C, photosynthesis in LT seedlings might be limited by Rubisco activase capacity, while in HT seedlings, acclimation removed this limitation.     

Two tertiary branched tetraamines, N4-aminopropylnorspermidine and N4-aminopropylsperimidine, are the major polyamines in thermophilic eubacteria, Thermoleophilum

Abstract Two novel polyamines were found as major polyamines of Thermoleophilum album and Thermoleophilum minutum , which are Gram-negative eubacteria obligate for thermophily and n -alkane substrates. They were identified as tertiary branched tetraamines, N⁴-aminopropylnorspermidine ( tris (3-aminopropyl)amine) [NH₂(CH₂)₃N-((CH₂)₃NH₂)(CH₂)₃NH₂ or N(CH₂CH₂CH₂NH₂)₃] and N⁴-aminopropylspermidine [NH₂(CH₂)₃N((CH₂)₃NH₂)(CH₂)₄NH₂] by high-performance liquid chromatography and gas chromatography-mass spectrometry.     

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.     

Properties of the ATP phosphohydrolase of the facultative thermophile Bacillus coagulans

Abstract The thermostability of the ATP phosphohydrolase of the facultative thermophile Bacillus coagulans has been investigated. Fractionation of disintegrated cell suspensions by differential centrifugation revealed a similar distribution of enzyme activity irrespective of growth temperature. Most of the activity was located in the membrane fraction. Thermostability of solubilized (BF₁) preparation from cells grown at 37°C or 55°C was similar, but membrane-bound BF₀BF₁ from 37°C-grown cells was inactivated at lower temperatures than that from 55°C-grown cells.
Inhibition of the membrane-bound (BF₀BF₁)ATPase by 4-chloro-7-nitro-benzofuran (NbfCl) and quercetin, which both act on the BF₁ portion of the enzyme, was different from that seen with the soluble (BF₁) enzyme. The results show that some modification of BF₁ must occur when the enzyme is membrane-bound.     

Mice with targeted disruption of spermidine/spermine N1-acetyltransferase gene maintain nearly normal tissue polyamine homeostasis but show signs of insulin resistance upon aging

The N¹-acetylation of spermidine or spermine by spermidine/spermine N¹-acetyltransferase (SSAT) is the ratecontrolling enzymatic step in the polyamine catabolism. We have now generated SSAT knockout (SSAT-KO) mice, which confirmed our earlier results with SSAT deficient embryonic stem (ES) cells showing only slightly affected polyamine homeostasis, mainly manifested as an elevated molar ratio of spermidine to spermine in most tissues indicating the indispensability of SSAT for the spermidine backconversion. Contrary to SSAT deficient ES cells, polyamine pools in SSAT-KO mice remained almost unchanged in response to N¹, N¹¹-diethylnorspermine (DENSPM) treatment compared to a significant reduction of the polymine pools in the wild-type animals and ES cells. Furthermore, SSATKO mice were more sensitive to the toxicity exerted by DENSPM in comparison with wild-type mice. The latter finding indicates that inducible SSAT plays an essential role in vivo in DENSPM treatmentevoked polyamine depletion, but a controversial role in toxicity of DENSPM. Surprisingly, liver polyamine pools were depleted similarly in wild type and SSAT-KO mice in response to carbon tetrachloride treatment. Further characterization of SSAT knockout mice revealed insulin resistance at old age which supported the role of polyamine catabolism in glucose metabolism detected earlier with our SSAT overexpressing mice displaying enhanced basal metabolic rate, high insulin sensitivity and improved glucose tolerance. Therefore SSAT knockout mice might serve as a novel mouse model for type 2 diabetes.     

Induction of secondary dormancy in sunflower seeds by high temperature. Possible involvement of ethylene biosynthesis

High temperature (45°C) inhibits seed germinition and seedling sunflower ( Helianthus annuus L. cv. Mirasol). Treatment of imbibed seeds at 45°C for more than 48 h induces a secondary dormancy, which is associated with progressive decrease of germination ability at optimal temperature (25°C) as well as with abnormal seedling growth. Ethylene (55μl l⁻¹) and 2-chloroethylphosphonic acid (ethephon) (2.5 m M ) improve germination of thermodormant seeds at 25°C. but the abnormal growth of the seedlings remains. O₂-enriched atmosphere and dry storage improve germination and normal seedling growth. The induction of thermodormancy in sunflower seeds seems associated with loss of their ability to convert 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene. Possible effects of high temperature on membranes and ethylene forming enzyme (EFE) are discussed.     

Regulation of the loss of frost hardiness in Pinus radiata by photoperiod and temperature

Abstract. In controlled environments, the interactive effects of warm (16: 8°C, day: night) and cool (12: 4°C, day: night) temperatures and long (13.5 h) and short (10 h) photoperiods on the dehardening of seedlings of Pinus radiata D. Don were investigated. In another experiment, the effect of four photoperiods from 9 to 14 h was examined. In a third, dehardening at constant temperatures from 5 to 17°C was followed. There was no evidence for an interaction between photoperiod and temperature. Dehardening was temporarily delayed by photoperiods below about 10 h, but there was no other quantitative effect of photoperiod. At constant temperatures, the rate of dehardening was initially constant but declined as the minimum summer frost hardiness was reached. In the initial phase the rate of dehardening was a linear function of temperature, increasing from 0.05°C day⁻¹ at 8°C to 0.30 °C day⁻¹ at 17°C. Temperature controlled the loss of frost hardiness by regulating the rate of dehardening.     

Latitudinal cline of requirement for far‐red light for the photoperiodic control of budset and extension growth in Picea abies (Norway spruce)

To test for the effects of far‐red light on preventing budset in Picea abies , seedlings of six populations originating from latitudes between 67°N and 47°N were grown for 4–8 weeks in continuous incandescent (metal halogen) light at 300 µmol m⁻² s⁻¹ and 20°C and then transferred, at the same temperature, to a daily regime of 8 h incandescent light (300 µmol m⁻² s⁻¹) followed by 16 h cool white fluorescent light (40 µmol m⁻² s⁻¹). (Cool white lamps are deficient in far‐red light, with a R/FR ratio of 7.5 compared with 2.0 for the incandescent lamps.) All the seedlings from 67° and 80% of those from 64° stopped extension growth and set terminal buds within 28 days of the change of regime. The seedlings from 61° and further south continued growing, as did control seedlings from 67° grown as above but with incandescent light at 20 µmol m⁻² s⁻¹ replacing cool white illumination. To distinguish between a clinal and ecotypic pattern of variation, the interval between 64° and 59° was investigated by growing populations originating from that area in the same regimes as before. After 28 days in the cool white day‐extension regime, the percentage budset was 86 for the population from 64°, 0 for the population from 59° and 25–50 for the intermediate populations; i.e. the populations showed a clinal variation in requirement for far‐red light according to latitude. Thus northern populations of Picea abies appear to behave as 'light‐dominant' plants for the photoperiodic control of extension growth and budset, whereas the more southern populations behave as 'dark‐dominant' plants.     

Effect of different night temperature regimes on the assimilation, transport and metabolism of carbon in rose plants

A decrease in CO₂ uptake, an increase in leaf starch and sucrose content and a decrease in the content of reducing sugars were found in rose ( Rosa hybrida cvs: Sonia and Golden Times) plants exposed to lower night temperature (12°C) in comparison with those grown at 18°C. These responses were not present when plants were grown under a night temperature regime of alternating temperature, 2 h at 18°C followed by 2 h at 12°C for 3 repetitive periods of a total of 12 h. The export of labelled carbon from the source leaves and translocation into the adjacent axillary buds were inhibited by lower night temperature, but not by the alternating temperature regime. The translocation of labelled carbon toward the basal plant parts was promoted by the lower temperature but not by alternating temperature. The partitioning of labelled carbon between the 2 uppermost lateral buds was also affected by the night temperature regime.     

嫁接对铜胁迫下黄瓜幼苗根系多胺代谢的影响

采用营养液栽培法,研究了嫁接(以黑籽南瓜为砧木)对铜胁迫下黄瓜幼苗根系活力及多胺代谢的影响.结果表明:铜胁迫下黄瓜幼苗根系活力下降,电解质渗漏率升高,而嫁接苗的变化幅度显著小于黄瓜自根苗;铜胁迫下黄瓜嫁接植株根系中除游离态腐胺(Put)含量显著低于自根苗外,结合态和束缚态Put、3种形态亚精胺(Spd)和精胺(Spm)含量均显著高于自根苗,嫁接苗根系中游离态Put含量及腐胺/多胺(Put/PAs)显著低于自根苗;铜胁迫下,嫁接苗根系精氨酸脱羧酶(ADC)、鸟氨酸脱羧酶(ODC)和S-腺苷蛋氨酸脱羧酶(SAMDC)活性高于自根苗,而二胺氧化酶(DAO)和多胺氧化酶(PAO)活性显著低于自根苗.表明嫁接黄瓜幼苗根系PAs的合成增加,降解减少,使PAs含量维持在较高水平,从而提高了黄瓜幼苗抗铜胁迫能力.     

Characterization of phe B gene encoding catechol 2,3-dioxygenase

A shaken thermal gradient device providing temperatures between 8.3 and 33.5° C was used to investigate the effects of silver ion on the duration of the lag phase and on minimum apparent growth temperatures of Hyphomicrobium spp. grown at 29 and 9°C. With 29°C-grown inocula, at lower temperatures, an increased time was required for growth initiation in the presence of silver ion added at 5 ng ml^-1. With silver ion added at 10 or 100 ng ml^-1, growth initiation was not observed at lower temperatures. With 100 ng ml^-1 added silver ion, this effect also was observed with 9°C-grown inocula. This increased sensitivity to silver ion could limit the ability of Hyphomicrobium spp., and possibly other microbes, to initiate growth and to contribute to microbial functioning in silver-impacted low temperature environments.     

Gas exchange and carbon isotope composition of Ananas comosus in response to elevated CO2 and temperature

Ananas comosus L. (Merr.) (pineapple) was grown at three day/night temperatures and 350 (ambient) and 700 (elevated) μ mol mol^–1 CO₂ to examine the interactive effects of these factors on leaf gas exchange and stable carbon isotope discrimination ( Δ ,‰). All data were collected on the youngest mature leaf for 24 h every 6 weeks. CO₂ uptake (mmol m^–2 d^–1) at ambient and elevated CO₂, respectively, were 306 and 352 at 30/20 °C, 175 and 346 at 30/25 °C and 187 and 343 at 35/25 °C. CO₂ enrichment enhanced CO₂ uptake substantially in the day in all environments. Uptake at night at elevated CO₂, relative to that at ambient CO₂, was unchanged at 30/20 °C, but was 80% higher at 30/25 °C and 44% higher at 35/25 °C suggesting that phosphoenolpyruvate carboxylase was not CO₂-saturated at ambient CO₂ levels and a 25 °C night temperature. Photosynthetic water use efficiency (WUE) was higher at elevated than at ambient CO₂. Leaf Δ -values were higher at elevated than at ambient CO₂ due to relatively higher assimilation in the light. Leaf Δ was significantly and linearly related to the fraction of total CO₂ assimilated at night. The data suggest that a simultaneous increase in CO₂ level and temperature associated with global warming would enhance carbon assimilation, increase WUE, and reduce the temperature dependence of CO₂ uptake by A. comosus .     

THE COMBINED EFFECTS OF LOW TEMPERATURE AND SO2+NO2 POLLUTION ON THE NEW SEASON'S GROWTH AND WATER RELATIONS OF PICEA SITCHENSIS

Picea sitchensis (Bong.) Carr. seedlings were exposed to SO₂, NO₂ and SO₂+ NO₂ during dormancy in controlled environments, and were taken to night temperatures of 4, 0, −5, −10 and −15 °C in a freezer. Conditions in the freezer were carefully monitored during the low–temperature treatments. In two experiments, different photoenvironments and temperature regimes were imposed prior to the cold treatments, and different effects were observed. In the first, only limited frost hardiness was achieved and night temperatures of −15 °C were lethal. Temperatures of −5 and − 10 °C led to poor survival of lateral buds, particularly in plants exposed to 45 ppb SO₂. The poor bud break in plants exposed to SO₂ and to − 5 °C resulted in a loss of the effectiveness of this temperature as a chill requirement. Pressure-volume analysis showed that the shoots of plants exposed to NO₂ had greater elasticity (lower elastic moduli, e), so that loss of turgor occurred at lower relative water contents. In contrast, a hardening period (2 weeks in night/day temperatures of 3/10 °C and 8 h days at 50 μmol m⁻² s⁻¹ PAR) gave decreased elasticity and lower solute potentials of spruce shoots. In the second experiment, exposure to 30 ppb SO₂ and SO₂+ NO₂ led to slight, but consistent, increases in frost injury to the needles of plants frozen to − 5 and − 10 °C. The results suggest that the main interaction of low temperatures and winter pollutants may be on bud survival rather than on needle damage, but that effects are subtle, only occurring with certain combinations of pollutant dose and cold treatment.     

Stress responses of tobacco cells to high temperature and salinity. Proline accumulation and phosphorylation of polypeptides

Stress responses to high temperatures (35–48°C) and salinity (170–340 mM NaCl) and thermotolerance were investigated for the salt-sensitive wild type and the salt-tolerant N_rE_s-1 strain of Nicotiana sylvestris L. using suspension-cultured cells. Under saline conditions, N_rE_s-1 strain cells accumulated proline, polyamine (putrescine and spermidine) and betaine in contrast to wild-type cells. The simultaneous treatment of salt-tolerant cells with high temperature (40°C) and NaCl (170 or 340 mM ) resulted in a transient overproduction of proline accompanied by an increase in their thermotolerance. At the high temperature, the synthesis of polypeptides and the accumulation of heat shock protein HSP70 mRNA were not affected by salinity. The higher thermotolerance of the NaCl-tolerant cells could not be related to osmoprotecting sugar-starch interconversions but could rather be associated with selective phosphorylation of several polypeptides (23–24, 27, 31–32, 47 kDa) prior to the accumulation of proline. The possible role of polyamines and polypeptide phosphorylation in this respect is discussed.     

Mobilisation of reserves and the earliest synthesis of sterols and latex triterpenes during germination and early seedling growth of Euphorbia lathyris

Seedlings of Euphorbia lathyris L. were grown in the dark at 25°C. Levels of amino acids, sugars and soluble phosphate in the endosperm increased upon germination after 4 days of imbibition, while the amounts of mineral reserves Mg²⁺ and K⁺ started decreasing. Ca²⁺ was not translocated from the endosperm to the seedling. Maximum values for amino acids were found on day 7, and the highest amounts of sugars were present on day 10. The endosperm was completely depleted by day 12.
Before germination (days 1–3) a low level of sterol synthesis in the embryo was detected with labeled sucrose and serine and to a lesser extent with labeled pyruvate. The label of [2-¹⁴C]-mevalonate proceeded exclusively to squalene. Laticifers started the synthesis of their triterpenes upon germination (day 4), using sucrose as a main substrate. A concurrent increase of sterol synthesis outside the laticifers was traced with labeled serine. Radioactive triterpenes, 4 α-methylsterols and sterols were detected in growing seedlings after [¹⁴C]-mevalonate uptake, but most of its label accumulated in squalene. The use of labeled mevalonate in sterol synthesis in growing seedlings is discussed.     

Effect of α-tocopherol on growth, membrane-bound adenosine triphosphatase activity of the roots, membrane fluidity and potassium uptake in rice plants

The effect of application of α-tocopherol on the physiology of young rice seedlings ( Oryza sativa L. cv. Dunghan Shali) was studied. Application of α-tocopherol resulted in a growth stimulation of the shoots of 17.5% at the suboptimal 13°/8°C day/night temperature. Growth stimulation was accompanied by stimulation of K⁺-uptake of the roots and by stimulation of the Ca²⁺-stimulated ATPase activity of the microsomal fraction of the root cell membranes at low temperature in such a way that the non-linearity ('broken curve') of the Arrhenius plot of control plants of this chilling-sensitive species was absent and replaced by a linear relation, as in chillingresistant species. Application of α-tocopherol also resulted in increased membrane fluidity, and the data are explained as a structural disordering effect of α-tocopherol on the cell membranes of the rice seedlings.