Thermal shock and germination in North‐West European Genisteae: implications for heathland management and invasive weed control using fire

Question: Is the stimulation of germination by thermal shock (resulting from the passage of fire) commonly observed for Mediterranean‐climate Fabaceae also apparent for NW European Genisteae (Fabaceae) species? Location: Southern England and NW France. Methods: The germination of Cytisus scopiarius, Genista anglica, Ulex europaeus, Ulex gallii and Ulex minor was examined following exposure to a range of temperatures (50°C, 65°C, 80°C, 95°C and 110°C) applied to seeds for 5 min. A sixth Mediterranean‐origin species (Spartium junceum) was also included since it is a common invasive in NW Europe and North America. Results: All five native NW European species displayed increased germination following thermal shock, even when seeds were heated to 110°C. However, there was some variation depending on provenance: in contrast to seeds collected from southern England, germination of French C. scopiarius seeds was unaffected by temperature. Spartium junceum germinated most at 95°C, but was the only species to show reduced germination when seeds were heated to 110°C. Conclusions: The NW European Genisteae appear to be pre‐adapted to the high temperatures associated with fire; a response attributable to their evolutionary origins in the fire‐prone ecosystems of the Mediterranean Basin. Consequently, projected increases in fire frequency linked to climate change may stimulate their regeneration in NW European heathlands, potentially altering the species composition of these ecosystems. Additionally, a clearer understanding of the interaction between thermal shock and germination may explain why fire has so frequently been ineffective in controlling invasive Genisteae populations worldwide.     

Effect of acute pH depression on the survival of the freshwater amphipod Hyalella azteca at variable temperatures: field and laboratory studies

Field observations on temperature and pH of a small pond showed that a amphipod population of Hyalella azteca was exposed to variable seasonal pH between 5.10–5.85, and water temperatures between 2–21 °C. Laboratory experiments were designed to simulate seasonal temperatures and field pHs of a small pond habitat. Laboratory bioassay experiments were conducted to determine the survival of Hyalella azteca at pHs 4, 5, 6 and 7, and varying temperatures of 5°, 10°, 15°, 20° and 25 °C.The LT₁₀₀ at pH 4 and 25 °C was 5.7 ± 0.47 days, compared to 47.3 ± 2.49 days at 5 °C. An Analysis of Variance (ANOVA) showed temperature was a significant (p > 0.0001) source of variation in the acute lethality of pH to H. azteca. A Duncans Multiple Range Test (DMRT) further showed that in laboratory experiments at pH 4, there was a significant difference ( = 0.01) between the LT_100s at 5°, 10°, 15° and 20 °C, but not between temperatures 20° and 25 °C.     

Effect of temperature on infection, development and reproduction of the parasitic nematode Thripinema nicklewoodi in Frankliniella occidentalis

The parasitic nematodeThripinema nicklewoodi Siddiqi (Tylenchida:Allantonematidae) is currently underinvestigation for use in inoculative releasestrategies against western flower thrips (WFT),Frankliniella occidentalis Pergande(Thysanoptera: Thripidae) infesting greenhousefloricultural crops. The aim was to determinewhether temperatures within greenhouses wouldpermit the establishment of T.nicklewoodi. The abilities of T.nicklewoodi to infect, develop and reproducein WFT were assessed under a range of constantand fluctuating temperatures in the laboratory.At constant temperatures, T. nicklewoodiinfected WFT over the range of 1–30 °C,although the temperature-related infectionprofile followed an asymmetric distributionaround an optimum 20 °C (80%infection). The lower and upper thresholds forT. nicklewoodi in vivo development andreproduction were higher than for infection, at10 °C and 35 °C, respectively.Climate data recorded over 1999–2000 in acommercial greenhouse (Texas) revealed atemperature range of 15 °C to31 °C from early March through mid June,when WFT were most abundant. While low(nighttime) greenhouse temperatures areconducive for T. nicklewoodi, upperdaytime temperatures are close to the upperthreshold for infection and may reducereproductive output. However, fluctuatingtemperature bioassays in the laboratorydemonstrated that T. nicklewoodimaintained separately at the upper thresholdtemperatures for infection (30 °C) anddevelopment (35 °C) readily infected anddeveloped in WFT when they were allowedintermittent (10 h daily) exposure to apermissive temperature in the range10–20 °C. Drawing on the results, thediurnal temperature-fluctuations of variousgreenhouses growing ornamentals would permitthe establishment of T. nicklewoodi.     

Influence of heat on seed germination of seven Mediterranean Leguminosae species

The influence of high temperatures (dry heat and hot water) on germination of seven Mediterranean Leguminosae species typical of fire-prone ecosystems in southern Spain is analyzed, in order to know the response of seeds to wildfires and the possible implications in their regeneration after this disturbance. Seeds were heated to a range of temperatures (50 °–150 °C) and exposure times (1–60 min) similar to those registered in the upper soil layers during wildfires. Germination tests were carried out in plastic Petri dishes over 60 days. In general, the degree of seed germination promotion by dry heat treatments showed a wide interspecific variation, although the final germination level was increased in all the studied species except for Scorpiurus muricatus. The thermal pretreatment of 50 °C, however, was not effective for germination in any species, and rising the temperature to 70 °C only slightly enhanced the germination in Cytisus patens. The preheatings of 90 °C (5 and 10 min), 120 °C (5 and 10 min), and 150 °C (1 min) were the most effective in promoting seed germination. Hot water (100 °C) scarification also increased the final germination level in all cases, with the exception of C. patens. The germination rates after preheating were much lower than in mechanically scarified seeds and closely resembled those of the untreated seeds, except for C. reverchonii, whose seed germination rate decreased with heat. The response of species to heat shock had no clear relationship with life trait or with the specific post-fire regeneration strategy (obligate seeder or facultative resprouter). Those species coexisting in the same habitats had different heat optimal requirements for seed germination, an strategy suggested by some authors as minimizing interspecific competition in the secondary succession started after fire.     

Adaptive response to cold temperatures and characterization of cspA in Salmonella typhimurium LT2

Salmonella typhimurium is a major foodborne microbial pathogen which primarily contaminates poultry products causing salmonellosis in humans. S. typhimurium LT2 cultures, when transferred from 37 °C to 5 °C or 10 °C, showed an initial lag period in growth with an approximate generation time of 10–25 h. Western blot assay using E. coli CS7.4 antibody and analysis of radiolabeled total cellular proteins from S. typhimurium cultures after exposure to 10 °C or 5 °C showed elevated expression of a major cold shock protein, CS7.4. Identification of a decreased level of CS7.4 at 37 °C suggests that the expression of this protein may require a large temperature downshift. Putative regulatory protein binding segment on the 5-untranslated region referred as Fragment 7 in S. typhimurium exhibited a 90.6% and a 56.25% nucleotide sequence identity when compared with the Fragment 7 of E. coli and S. enteritidis, respectively. The differences in the nucleotide sequence within the Fragment 7 between S. typhimurium and S. enteritidis may explain the differential expression of CspA at 37 °C. The nucleotide sequence of the open reading frame of S. typhimurium cspA gene showed a single base difference at 816 bp position from a G to a C which altered the amino acid residue from a glycine to an alanine. In addition to CspA, an elevated expression of a 105 kDa, and decreased expression of 6 proteins were evidenced when cultures of S. typhimurium were exposed to 10 °C or 5 °C. Differential expression of the CspA and other proteins in S. typhimurium following exposure to cold temperatures suggest that adaptation and continued growth and survival at cold temperatures in this pathogen may be aided by these cold-responsive proteins.     

Effect of abscisic acid, osmolarity and temperature onin vitro development of recalcitrant mango nucellar embryos

Development of cotyledonary-stage nucellar embryos of mango was arrestedin vitro by exposure to 750–1750 M ABA. The enlargement and germination of nucellar embryos was inhibited for as long as 4 weeks after subculture from ABA-containing medium. Mannitol at concentrations between 7.5 and 12.5% inhibited nucellar embryo development, presumably due to osmotic effects; however, there was no residual effect after subculture of somatic embryos onto medium without mannitol. Temperatures between 22.5 and 37.5°C stimulated embryo development, whereas lower temperatures (7.5 and 15°C) delayed germination. There was no germination 1 month after somatic embryos, pulsed for 8 weeks at 7.5°C, were transferred to 22.5°C; however, after 2 months, 86% of these somatic embryos germinated. These results indicate that it is possible to induce developmental arrest in recalcitrant mango embryos with high concentrations of ABA, mannitol or low temperature (7.5°C).Abbreviations ABA Abscisic acid - MM1 Mango maturation medium     

The relationship between fire history and an exotic fungal disease in a deciduous forest

Exotic diseases have fundamentally altered the structure and function of forest ecosystems. Controlling exotic diseases across large expanses of forest has proven difficult, but fire may reduce the levels of diseases that are sensitive to environmental conditions. We examined Cornus florida populations in burned and unburned Quercus–Carya stands to determine if burning prior to anthracnose infection has reduced the impacts of an exotic fungal disease, dogwood anthracnose, caused by Discula destructiva. We hypothesized that fire has altered stand structure and created open conditions less conducive to dogwood anthracnose. We compared C. florida density, C. florida health, and species composition and density among four sampling categories: unburned stands, and stands that had burned once, twice, and 3 times over a 20-year period (late 1960s to late 1980s). Double burn stands contained the greatest density of C. florida stems (770 stems ha⁻¹) followed by triple burn stands (233 stems ha⁻¹), single burn stands (225 stems ha⁻¹) and unburned stands (70 stems ha⁻¹; P < 0.01). We observed less crown dieback in small C. florida trees (<5 cm diameter at breast height) in burned stands than in unburned stands (P < 0.05). Indicator species analysis showed that burning favored species historically associated with Quercus–Carya forests and excluded species associated with secondary succession following nearly a century of fire suppression. Our results suggest that fire may mitigate the decline of C. florida populations under attack by an exotic pathogen by altering forest structure and composition. Further, our results suggest that the burns we sampled have had an overall restorative effect on forest communities and were within the fire return interval of the historic fire regime. Consequently, prescribed fire may offer a management tool to reduce the impacts of fungal disease in forest ecosystems that developed under historic burning regimes.     

Characterization of the heat shock response inEnterococcus faecalis

We have characterized the general properties of the heat shock response of the Gram-positive hardy bacteriumEnterococcus faecalis. The heat resistance (60°C or 62.5°C, 30 min) of log phase cells ofE. faecalis grown at 37°C was enhanced by exposing cells to a prior heat shock at 45°C or 50°C for 30 min. These conditioning temperatures also induced ethanol (22%, v/v) tolerance. The onset of thermotolerance was accompanied by the synthesis of a number of heat shock proteins. The most prominent bands had molecular weights in the range of 48 to 94kDa. By Western blot analysis two of them were found to be immunologically related to the well known DnaK (72 kDa) and GroEL (63 kDa) heat shock proteins ofEscherichia coli. Four other proteins showing little or no variations after exposure to heat are related to DnaJ, GrpE and Lon (La)E. coli proteins and to theBacillus subtilis ⁴³ factor. Ethanol (2% or 4%, v/v) treatments elicited a similar response although there was a weaker induction of heat shock proteins than with heat shock.     

Heat effects on seeds and rhizomes of a selection of boreal forest plants and potential reaction to fire

To analyse the potential reaction to firegenerated heat pulses, seeds of 12 species of plants and rhizomes of three species were exposed to elevated temperatures for 10 min. The tested material split into three groups with respect to heat tolerance: (1) the rhizomes, for which the lethal temperatures were in the range 55–59° C; (2) the seeds of most of the species tested, for which the lethal temperatures were in the range 65–75° C; (3) The seeds of two species of Leguminosae and three species of Geranium for which the lethal temperatures were around 100° C. For all three Geranium species and for one of the legume species, Anthyllis vulneraria, exposure temperatures above ca. 45° C resulted in dormancy release, and maximum germination occurred above 60–65° C. Speed of germination was little affected for most species, except after exposure to nearlethal temperatures, where it slowed down dramatically, although the seedlings emerging were healthy. We conclude that due to sharp temperature gradients in the soil during fire, differences in heat tolerance between species in most cases are not large enough to be a decisive factor in their post-fire colonising success. There are exceptions: the seeds of certain taxa that are impermeable to water in the dormant state, some of which have heat triggered germination.     

Induction of secondary dormancy in Amaranthus caudatus seeds

Nondormant A. caudatus seeds germinated in the darkat temperatures between 20 and 35° but not at 45 °C.Incubation at this temperature for at least 10 h inhibited seedgermination over the temperature range 20 to 35 °C,temperatures previously suitable for germination. Thus incubation at 45°C induced secondary dormancy. Mechanical or chemicalscarification or exposure to pure oxygen caused complete or almost completegermination of dormant seeds although more slowly in comparison to nondormantseeds. Secondary dormant scarified seeds required a lower concentration of ABAthan nondormant seeds to inhibit germination. The high temperature, whichinduced dormancy, 45 °C, caused the seed coat to be partiallyresponsible for secondary dormancy. Involvement of ABA (synthesis orsensitivity) in the induction and/or maintenance of this dormancy should beconsidered.     

Shrub invasion into subalpine vegetation: implications for restoration of the native ecosystem

The ability of plant communities to recover after non-native species invasion will depend upon the nature of their soil seed bank and seed rain characteristics. This study assessed changes in the soil seed bank and seed rain associated with the invasion of the non-native shrub Cytisus scoparius in subalpine vegetation. Soil seed bank and seed rain composition, density and richness were investigated at three areas of different stages of invasion: (i) recent (8–10 years), (ii) mature (15–16 years) and (iii) long-term (25 years). There were few changes in seed bank composition or richness regardless of invasion stage. By contrast, the seed rain composition, richness and density was substantially different within long-invaded areas. Very few seeds were able to colonise the dense barrier characteristic of larger, more mature C. scoparius stands. Some prominent herbs from the native vegetation were under-represented or absent from the seed bank, both in invaded and uninvaded areas. Laboratory germination experiments demonstrated that most native species germinate easily, which may imply a transient seed bank, rather than a persistent one. The majority of herbaceous and shrub species were capable of resprouting vegetatively. Therefore, regeneration appeared more reliant on the bud and tuber bank than a persistent soil seed bank. The dominance of graminoid species and C. scoparius rather than other herbaceous, shrub or tree species suggests that the regenerating vegetation will be dominated by grass species and/or C. scoparius. Hence, in areas where long-invaded C.␣scoparius stands are present the recovery of native subalpine vegetation maybe difficult. Recovery may only be possible through wind dispersal from the surrounding intact vegetation or through actively reseeding the area. This study highlights the importance of early intervention in invasive species management.     

Characterisation of Soybean and Wheat Seeds by Nuclear Magnetic Resonance Spectroscopy

The effects of equilibration under different air relative humidities (RH, 1 – 90 %) and temperatures (35 and 45 °C) on soybean (Glycine max) and wheat (Triticum aestivum) seeds were studied using different techniques. Seed moisture content, electrical conductivity (EC) of seed leachate and per cent seed germination were measured following standard procedures, and compared with nuclear magnetic resonance spin-spin relaxation time (T₂) measurements. Moisture contents of soybean and wheat seeds, following the reverse sigmoidal trend, were greater at 35 than at 45 °C at any particular RH. Changes in T₂ were related to the changes in germination percentage and leachate EC of both soybean and wheat seeds. Equilibrating soybean seeds at RH 11 % decreased germination percentage with corresponding decrease in T₂. On the contrary, EC of seed leachate increased. In wheat seeds equilibrated at 45 °C, T² was maximal at RH 5.5 %. T₂ declined in seeds equilibrated at high RH (> 80 %) together with low germination percentage.     

Germination responses of <Emphasis Type="Italic">Caragana korshinskii</Emphasis> Kom. to light,temperature and water stress

Caragana korshinskii Kom. is a very important shrub species for vegetation rehabilitation in northern China for its high ecological and economic values. Experiments were conducted to determine its germination responses to (i) different temperature regimes under light and/or dark conditions, (ii) different light intensities, and (iii) different water potentials combined with varied constant temperatures. Under alternating temperatures (from 5:15 to 25:35°C), final percent germinations of Caragana korshinskii were quite similar. In dark conditions, constant temperatures resulted in lower final percent germinations than alternating temperatures. At a controlled temperature regime of 10:20°C, neither final percent germinations nor germination rates showed significant differences among varied light intensities. As water potentials were reduced from 0 (distilled water) to –0.6MPa, final percent germinations increased slightly and reached the peak at approximately –0.6MPa, however, the increment was not significant. Beyond –0.6MPa, further water potential reduction led to decreased final percent germinations and few seeds could germinate at –1.4MPa. Water stress also strongly inhibited germination at very high or low temperatures. The experimental results suggested that middle May might be a suitable time for aerial seeding for this species.     

Survival of the gemmules of Anheteromeyenia ryderi (Potts) following aerial exposure during winter in New England

Gemmules of Anheteromeyenia ryderi survived 24 h exposure to air temperatures as low as –20 °C under laboratory conditions. Drying the gemmules of A. ryderi at 5 °C under laboratory conditions resulted in a reduced viability and a slower germination rate following rehydration compared with undried control gemmules. Only 25% of the gemmules germinated after drying for one month. Up to 25% of the gemmules of A. ryderi that were tested survived aerial exposure from early November to early April when a pond in Connecticut inhabited by this sponge was drained. During this period air temperatures dropped to as low as –16 °C. Continued aerial exposure of the gemmules during the summer resulted in nearly complete gemmule mortality.     

The effect of ethrel and temperature on the germination of Manihot glaziovii Muell. Arg. seeds

Dormancy in seeds of Manihot glaziovii is overcome at 25°C by application of ethrel at effective ethylene concentrations equal to and greater than 10 ll^–1. Imbibition of seeds in ethrel broadens the temperature optimum for germination but does not prevent the development of secondary dormancy at temperatures of 35°C and greater and 15°C and lower. Secondary dormancy must be due to factors other than reduced ethylene production.     

The CO2/O2 specificity of ribulose 1,5-bisphosphate carboxylase/oxygenase

The substrate specificity factor, V _cK_o/V_oK_c, of spinach (Spinacia oleracea L.) ribulose 1,5-bisphosphate carboxylase/oxygenase was determined at ribulosebisphosphate concentrations between 0.63 and 200 M, at pH values between 7.4 and 8.9, and at temperatures in the range of 5° C to 40° C. The CO₂/O₂ specificity was the same at all ribulosebisphosphate concentrations and largely independent of pH. With increasing temperature, the specificity decreased from values of about 160 at 5° C to about 50 at 40° C. The primary effects of temperature were on K _c [Km(CO₂)] and V _c [Vmax (CO₂)], which increased by factors of about 10 and 20, respectively, over the temperature range examined. In contrast, K _o [Ki (O₂)] was unchanged and V _o [Vmax (O₂)] increased by a factor of 5 over these temperatures. The CO₂ compensation concentrations () were calculated from specificity values obtained at temperatures between 5° C and 40° C, and were compared with literature values of . Quantitative agreement was found for the calculated and measured values. The observations reported here indicate that the temperature response of ribulose 1,5-bisphosphate carboxylase/oxygenase kinetic parameters accounts for two-thirds of the temperature dependence of the photorespiration/photosynthesis ratio in C₃ plants, with the remaining one-third the consequence of differential temperature effects on the solubilities of CO₂ and O₂.Abbreviations RuBPC/O(ase) ribulose 1,5-bisphosphate carboxylase/oxygenase - RuBP ribulose 1,5-bisphosphate - CO₂ compensation concentration     

A highly thermostable ferritin from the hyperthermophilic archaeal anaerobe Pyrococcus furiosus

A ferritin from the obligate anaerobe and hyperthermophilic archaeon Pyrococcus furiosus (optimal growth at 100°C) has been cloned and overproduced in Escherichia coli to one-fourth of total cell-free extract protein, and has been purified in one step to homogeneity. The ferritin (PfFtn) is structurally similar to known bacterial and eukaryal ferritins; it is a 24-mer of 20 kDa subunits, which add up to a total Mr 480 kDa. The protein belongs to the non-heme type of ferritins. The 24-mer contains approximately 17 Fe (as isolated), 2,700 Fe (fully loaded), or <1 Fe (apoprotein). Fe-loaded protein exhibits an EPR spectrum characteristic for superparamagnetic core formation. At 25°C V_max=25 mole core Fe³⁺ formed per min per mg protein when measured at 315 nm, and the K_0.5=5 mM Fe(II). At 0.3 mM Fe(II) activity increases 100-fold from 25 to 85°C. The wild-type ferritin is detected in P. furiosus grown on starch. PfFtn is extremely thermostable; its activity has a half-life of 48 h at 100°C and 85 min at 120°C. No apparent melting temperature was found up to 120°C. The extreme thermostability of PfFtn has potential value for biotechnological applications.     

Safe Site and Seed Limitation in Cytisus Scoparius (Scotch Broom): Invasibility, Disturbance, and the Role of Cryptogams in a Glacial Outwash Prairie

In all plant populations, establishment is controlled by two factors: the supply of propagules and their access to ‘safe sites’ for growth. An infestation of invading pest plants results in a seed-production gradient, from the edge where seeds are limiting, to the center where seeds may be in excess. Do invaded sites become ‘saturated’ with seeds? How rapidly does this occur, and how does the process depend on the availability of safe sites? Are safe sites, and consequently invasion, promoted by disturbance? I quantified the response of seedling establishment to seed input and disturbance in Cytisus scoparius (Scotch broom), an exotic shrub invading glacial outwash prairie remnants in western Washington, USA. I used disturbance treatments to investigate the role of the thick cryptogamic layer in these prairies, disturbing cryptogams by scraping or by fire. The effect of fire was partitioned into two factors: burning of the background vegetation/substrate versus breaking C. scoparius seed dormancy, by adding seeds either before or after the burn. Seed treatments ranged from 20 to 1000 seeds per m². Both seed number and surface treatment showed significant effects on seedling density, along with a significant interaction between the two factors. Disturbance did not promote C. scoparius establishment; undisturbed plots produced more seedlings than burned or scraped plots. Within the burned plots, fire scarification appeared to increase germination but this effect was not significant. For germinated seedlings, mortality through the dry season (June–August) was not significantly different among surface treatments, nor did survivorship depend on density, with the result that initial differences in germination among the treatments persisted. The message that the undisturbed cryptogam layer facilitates C. scoparius establishment suggests that ‘ecosystem management’ strategies promoting healthy, undisturbed sites will not always be effective against invasive pest species.     

Altered development of polysomal RNA activity in chick-pea (Cicer arietinum L.) embryonic axes. Effects of abscisic acid and temperature

The in vitro activity of polysomal polyadenylated RNA (poly(A)RNA) was studied using chick-pea (Cicer arietinum L.) embryonic axes subjected to treatments retarding germination (H₂O 30°C and abscisic acid [ABA] 30°C) or inducing a false germination (thiourea 30°C) in which normal protein synthesis and growth did not occur. All treatments induced a smaller proportion of poly(A)RNA compared with the control (H₂O 25°C). However, poly(A)RNA obtained in the presence of ABA had a similar in vitro activity to that of the control. The translation of mRNA from embryonic axes germinated at high temperatures was extensively blocked (70%) by methyl-7-guanosine-5-triphosphate, whereas mRNA translation from axes treated with H₂O-25°C and ABA was completely blocked (100%), indicating a greater cap dependence in the latter cases. Polyacrylamide gel electrophoresis showed that ABA and H₂O-30°C each induced the synthesis of a polypeptide with an approximate M_r of 32 kDa, probably a germination regulator. It is suggested that ABA and high temperatures could regulate germination at the translational level as well as affecting ionic-exchange properties, as has been previously demonstrated (Hernández-Nistal et al. 1983, Physiol. Plant. 57, 273–278).Abbreviations ABA abscisic acid - Poly (A) RNA polyadenylated RNA - TU thiourea     

Responses of florence fennel (Foeniculum vulgare azoricum) seeds to light,temperature and gibberellin A4/7

The percentage and the rate of germination of seeds of three varieties of Florence fennel was higher in the dark than in the light, the high temperature cut-off points being between 27.2 and 29.4°C. The optimum temperature for germination was between 20 and 25°C. Seeds of all three varieties responded to incubation in solutions containing gibberellin A_4/7 mixture (GA_4/7; Regulex), giving higher germination in the light at temperatures from 20 to 30°C. Seeds steeped for 4 h at 25°C or for 24 h at 5°C in GA_4/7 solutions gave a higher percentage and increased rate of emergence as compared with untreated dry seeds, when sown in compost at 25°C; steeping in water alone was also beneficial. In general, drying the treated seeds before sowing reduced the rate but sometimes increased the percentage of germination as compared with seeds sown when still moist. Seeds harvested from secondary umbels of var. Zefa fino germinated better both in the light and dark than those taken from primary umbels.Abbreviations GA_4/7 a commercial mixture of the gibberellins A₄ and A₇ (trade name Regulex)