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)     

Effect of high temperatures on seed germination of two woody Leguminosae

Cytisus scoparius and Genista florida regenerate after fire by stump-sprouting but also by seed. Seeds of these species were heated to a range of temperatures similar to those registered on the surface soil during natural fires (from 50 to 150 °C) and a range of exposure times (from 1 to 15 min). No germination was observed at high temperatures, 130 °C, when the exposure time was 5 min or more. However, moderate heat treatments (at 70 and 100 °C) significantly increased the rate of germination relative to controls. Cytisus scoparius is more favoured by fire action than Genista florida, with germination rates slightly greater following 100 °C for 5 min and 130 °C for 1 min than after mechanical scarification.     

Dormancy of<Emphasis Type="Italic"> Arabidopsis</Emphasis> seeds and barley grains can be broken by nitric oxide

Seeds of Arabidopsis thaliana (L.) Heynh. and grains of barley (Hordeum vulgare L.) were used to characterize the affects of nitric oxide (NO) on seed dormancy. Seeds of the C24 and Col-1 ecotypes of Arabidopsis are almost completely dormant when freshly harvested, but dormancy was broken by stratification for 3 days at 4°C or by imbibition of seeds with the NO donor sodium nitroprusside (SNP). This effect of SNP on dormancy of Arabidopsis seeds was concentration dependent. SNP concentrations as low as 25 M reduced dormancy and stimulated germination, but SNP at 250 M or more impaired seedling development, including root growth, and inhibited germination. Dormancy was also reduced when Arabidopsis seeds were exposed to gasses that are generated by solutions of SNP. Nitrate and nitrite, two other oxides of nitrogen, reduced the dormancy of Arabidopsis seeds, but much higher concentrations of these were required compared to SNP. Furthermore, the kinetics of germination were slower for seeds imbibed with either nitrate or nitrite than for seeds imbibed with SNP. Although seeds imbibed with SNP had reduced dormancy, seeds imbibed with SNP and abscisic acid (ABA) remained strongly dormant. This may indicate that the effects of ABA action on germination are downstream of NO action. The NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3 oxide (cPTIO) strengthened dormancy of unstratified and briefly stratified Arabidopsis seeds. Dormancy of three cultivars of barley was also reduced by SNP. Furthermore, dormancy in barley grain was strengthened by imbibition of grain with cPTIO. The data presented here support the conclusion that NO is a potent dormancy breaking agent for seeds and grains. Experiments with the NO scavenger suggest that NO is an endogenous regulator of seed dormancy.Abbreviations ABA Abscisic acid - cPTIO 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3 oxide - GA Gibberellin - SNP Sodium nitroprusside - NO_x Gaseous oxides of nitrogen     

Development and pigmentation of chlorosomes in Chloroflexus aurantiacus strain Ok-70-fl

The development of chlorosomes and their pigmentation were studied by growing Chloroflexus aurantiacus strain Ok-7o-fl first under conditions under which BChl c-synthesis is low (50°C, 2000 lux and 30°C, 1500 lux) and subsequently under conditions promoting high BChl c-synthesis (50°C, 400 lux). Electron microscopic observations on and chemical analyses of isolated cell components showed that in BChl c-depleted cells chlorosome-like structures (chlorosome bags) are attached to fragments of cytoplasmic membranes. These chlorosome bags exhibit a periodic fine structure caused by the construction of the baseplates of the chlorosomes. The baseplates are closely attached to the cytoplasmic membrane, they are rich in phospholipids and apparently contain a 790 nm-BChl a-complex. Chlorosome bags of BChl c-depleted cells always contain a limited amount of light-harvesting pigment complexes (BChlc, - and -carotene). The light-harvesting system is restored (50°C, 400 lux) by first refilling the existing chlorosome bags before cell division takes place.Abbreviations BChl Bacteriochlorophyll - LH Light-harvesting complex - RC Reaction center     

Responses of dormant heather (Calluna vulgaris) seeds to light, temperature, chemical and advancement treatments

Two seed lots of Calluna vulgaris were obtainedfrom English Nature (seed of Cornish provenance) (EN) and John ChambersWildflower Seeds (JCWS). In laboratory tests, under continuous light untreatedseeds of both seed lots were partially dormant at temperatures between14–35 °C, but JCWS seeds were more deeply dormant thanENseeds. The optimum temperature for germination for both lots was ca 18°C. Germination of EN seeds was much lower in the dark than inthe light at all temperatures; JCWS seeds did not germinate in the dark. In thelight at 22 °C, dormancy of both seed lots was broken whenseeds were incubated in GA_4/7 solution(2×10^–4 M). Dormancy ofJCWSseeds at 22 °C in the light was broken when seeds wereincubated in four different smoke solutions but more so when used incombinationwith GA_4/7. Soaking seeds for 4h insmoke/GA_4/7solutions before sowing improved both the speed andpercentage germination in pot experiments on a mist bench in the glasshouse byat least 10-fold. Soaking with GA_4/7 alone produced a 5-fold increasein germination but seedlings were more etiolated than with thesmoke/GA_4/7 mixtures. A seed advancement treatment modified from thatused commercially on sugar beet seeds also promoted germination in bothlaboratory and glasshouse tests. This entailed soaking seeds in 0.2% thiramsuspension for 4h followed by incubation in excess solution at 22°C for 4 days. This treatment was not as effective as thesmoke/GA_4/7 seed soaks.     

Feeding Ecology of Black-headed Uacaris (Cacajao melanocephalus melanocephalus) in Pico da Neblina National Park,Brazil

From June 1994 to October 1995, I conducted a study of the feeding ecology of one group of black-headed uacaris (Cacajao melanocephalus melanocephalus) in Pico da Neblina National Park, Brazil, near the Venezuelan border (01°10N to 00°26S, 65°03W to 66°52W). Of a total of 429 feeding records, 89% corresponded to fruits from which black-headed uacaris ate ripe and unripe seeds, mesocarps and arils. Seeds were the single most important food item year-around, corresponding to 81% of the feeding records for fruits. Black-headed uacaris used 120 different plant species in 32 families as food. The 3 most important plant species in their diet, Micrandra spruceana, Eperua leucantha, and Hevea braziliensis (all trees), accounted for 38% of the feeding records for fruits. These species produced fruits that were protected by very hard green husks, were barochorous, did not have fleshy mesocarps, and were slow to mature, thus being available as unripe fruits for long periods. The second and third most important species in the diet of black uacaris, Eperua leucantha and Hevea braziliensis, were dominant in the forest: approximately 30% of trees 10 cm dbh sampled in a 2-ha botanical transect at the study site. I discuss the formation of large social groups of black-headed uacaris in Pico da Neblina National Park in light of the peculiar floristic composition of the study site.     

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.     

Overcoming seed dormancy in ex situ plant germplasm conservation programmes; an example in the endemic Argyranthemum (Asteraceae: Anthemideae) species from the Canary Islands

Germplasm of 21 diverse Argyranthemum taxa was collected from contrasting ecological zones in the Canary Islands. Seed dormancy was considerable in the majority of taxa. Extensive investigations, based on a germination test procedure algorithm for Asteraceae, with achenes from ray and disc florets of five contrasting taxa identified a procedure to promote full (85%) germination of the seeds from both ray and disc florets of all five taxa; viz, excision of the seeds from the achenes, followed by testing at 15°C with 2.6×10^-3 m GA₃ co-applied. Subsequent tests showed that this regime was effective in promoting full germination in seeds from both ray and disc florets of the remaining 16 taxa. The results are discussed in the context of ex situ plant germplasm conservation.     

Seasonal variations in the immune system of the tench,Tinca tinca (Cyprinidae): proliferative response of lymphocytes induced by mitogens

The proliferation of tench lymphocytes induced by mitogens was studied during the four seasons of the year. Fish were maintained under natural conditions of photoperiod and temperature (mean ± SD: 12±2°C in winter, 22±3°C in spring, 30±3°C in summer and 21±3°C in autumn). Cultures were performed in vitro at 22°C in all seasons and the results were compared. Subsequently, in seasons with extreme water temperatures, cultures in vitro were performed at the same temperature as that of the water (12°C in winter and 30°C in summer) and the results were compared seasonally at the seasonal temperature, i.e. at 22°C in spring, 30°C in summer, 22°C in autumn and 12°C in winter. Phytohemagglutinin, concanavalin A, lipolisaccharide from E. coli and pokeweed mitogen were used as mitogens. Studies performed at 22°C as assay temperature in all seasons showed profound seasonal changes: while in spring, summer and autumn the mitogenic response of lymphocytes to phytohemagglutinin, concanavalin A, lipolisaccharide from E. coli and pokeweed mitogen was very low, during winter the results obtained were significantly higher. However, when the assays were performed at the corresponding seasonal temperature the differences were not as pronounced between the different seasons, and the mitogenic responses of lymphocytes were found to be the lowest during the winter and the highest during the summer with all mitogens used. This fact suggests that immunosuppression occurs in winter and an immunostimulation occurs in summer. However, the higher response found in winter when assaying at 22°C suggests that this property of lymphocytes needs an assay temperature higher than the in vivo temperature in order to observe accurate mitogenic responses.Abbreviations Con A concanavalin A - cpm counts per minute - LPS E. coli lipolisaccharide - MS222 tricainemethane sulphonate - PBS phosphate-buffered saline - PHA phytohemagglutinin - PWM pokeweed mitogen - SI stimulation index     

Effects of pre-storage conditions on storage of in vitro cultures of Nephrolepis exaltata (L.) Schott and Cordyline fruticosa (L.) A. Chev.

In vitro cultures of Nephrolepis exaltata and Cordyline fruticosa were stored at 5°, 9° or 13°C, at a low irradiance (3–5 mol m^–2 s^–1) or in darkness. Prior to storage the cultures were subjected to 18°, 21°, 24° or 27°C and 15, 30 or 45 mol m^–2 s^–1 in a factorial combination.The optimal storage conditions for Nephrolepis were 9°C in complete darkness. These cultures were still transferable to a peat/perlite mixture at the end of the experimental period of 36 months.The optimal storage conditions for Cordyline were 13°C and a low light level (±3–5 mol m^-2 s^-1). When the pre-storage conditions were normal growth room conditions (24°C and 30 mol m^-2 s^-1), in vitro cultures could be stored for 18 months. With the most favourable pre-storage treatment (18°C and 15 mol m^-2 s^-1) some cultures still had green shoots after 36 months of storage, but did not survive transfer to peat/perlite.Pre-conditioning before storage was most favourable for Nephrolepis, and not that important, but still favourable, for Cordyline. There was an interaction between pre-storage temperature and pre-storage irradiance. For both species a high irradiance level was less favourable than a low irradiance level when combined with high growth room temperatures.Abbreviations BA 6-benzyladenine - IAA indole-3-acetic acid - NOA 2-naphthoxyacetic acid     

Response of β-Glucosidase to Fungal Infections in Seed, Ovary and Fruit

Localization and changes in the activity of -glucosidase were investigated in wheat caryopsis and glumes infected with Stagonospora nodorum as well as in lily ovaries and harvested tomato fruits both inoculated with Botrytis cinerea. It was established that the pathogen invasion caused splitting of wheat seed coat, xylem blocking in lily carpel and decay in tomato fruits. B. cinerea invasion evoked disorders of the embryogenesis accompanied by a decreased activity of -glucosidase in all ovules. The activity of the enzyme was not changed considerably in wheat seeds as the infection occurred in the late embryonal stages and the embryonal processes were not affected. In the seeds of harvested tomatoes distant from the invaded area the enzyme activity was not changed as well.     

Dispersal of Eucalyptus torelliana seeds by the resin-collecting stingless bee,Trigona carbonaria

Ants are the only group of invertebrates currently identified as significant dispersers of seeds, but we report here the dispersal of Eucalyptus torelliana seeds by bees. Fruits of E. torelliana produce resin which is collected by workers of the stingless bee Trigona carbonaria. Seeds adhere to resin in the workers' corbiculate and are transported to the nest. Workers transported seeds distances of more than 300 m from the parent tree and seeds at the nest were viable and capable of germination. Seeds were removed from the nests by workers and discarded away from the nest, and E. torelliana trees became established in the vicinity of colonies of T. carbonaria. Mellitochory may be a novel method of seed dispersal where bees are dispersers, and associated with fruits that produce resin as an attractant for bees.     

The temperature-dependent duration of development and parasitism of three cereal aphid parasitoids, Aphidius ervi, A. rhopalosiphi, and Praon volucre

Temperature dependencies were established for the egg-to-mummy and mummy-to-adult phases, for mummy mortality, and for parasitism of Aphidius ervi Haliday, Aphidius rhopalosiphi De Stefani-Perez, and Praon volucre (Haliday) (Hymenoptera, Aphidiidae), three parasitoids of Sitobion avenae (Fabricius) (Homoptera, Aphididae), at 8°C, 12°C, 16°C, 20°C, and 25°C on winter wheat (cv. Haven). A physiological model described temperature-dependent development over the full temperature range, whereas a linear model was fitted for data above 8°C and used to estimate the lower temperature thresholds and day-degrees (° D) required for development. The thresholds for A. ervi were 2.2°C for egg-mummy development and 6.6°C for mummy-adult development, those for A. rhopalosiphi were 4.5°C and 7.2°C, and those for P. volucre were 3.8°C and 5.5°C. The time to develop into mummies and adults differed significantly between the three species: A. ervi development into mummies required an average of 159 ° D, while development into adults took an average of 73 ° D. The corresponding average times required for A. rhopalosiphi and P. volucre to develop mummies were 124° D and 126° D, while their development into adults required an average of 70° D and 150° D, respectively. Mummy mortality was 25–35% at 8°C and less at the higher temperatures tested, but began to increase again at 25°C, showing a quadratic relationship between mortality and temperature. Parasitization was very low or, in the case of P. volucre, absent up to 12°C and thereafter increased with increasing temperature. The relationship between parasitization, recorded as percent aphids mummified, and temperature was linear at the temperatures tested and depended on species. A. ervisuperparasitized 11.1% aphids at 20°C and 16.6% aphids at 25°C, whereas superparasitism was low in A. rhopalosiphi and absent in P. volucre. From 16°C to 25°C the P. volucre sex ratio increased. For A. ervi and A. rhopalosiphi there was no trend with temperature, but at 20°C and 25°C it was close to even. Field data for 1996 and 1997 allowed for a comparison of actual and expected emergence of overwintering mummies. In both years, parasitoids were predicted to have emerged from overwintering mummies well in advance of the onset of aphid infestation, and more than a month earlier than the first parasitized aphids were found in winter wheat. Observations from trap plants in other crops supported the predictions of the models. Other factors that can affect biological control by cereal aphid parasitoids are discussed.     

Abscisic acid and mefluidide in the regulation of cold hardiness development in Solanum tuberosum L. potato

Plants of Solanum tuberosum L. potato do not cold acclimate when exposed to low temperature such as 5°C, day/night. When ABA (45 M) was added to the culture medium, stem-cultured plantlets of S. tuberosum, cv. Red Pontiac, either grown at 20°C/15°C, day/night, or at 5°C, increased in cold hardiness from –2°C (killing temperature) to –4.5°C. The increase in cold hardiness could be inhibited in both temperature regimes if cycloheximide (70 M) was added to the culture medium at the inception of ABA treatment. Cycloheximide did not inhibit cold hardiness development, however, when it was added to the culture medium 3 days after ABA treatment.When pot-grown plants were foliar sprayed with mefluidide (50 M), ABA content increased from 10 nmol to 30 nmol g^–1 dry weight and plants increased in cold hardiness from –2°C to about –3.5°C. The increases in free ABA and cold hardiness occurred only in plants grown at 20°C/15°C; neither ABA nor cold hardiness increased in plants grown at 5°C.The results suggest that an increase in ABA and a subsequent de novo synthesis of proteins are required for the development of cold hardiness in S. tuberosum regardless of temperature regime, and that the inability to synthesize ABA at low temperature, rather than protein synthesis, appears to be the reason why S. tuberosum does not cold acclimate.     

The respiratory chain of the facultative thermophile,Bacillus coagulans

Two oxidases were found to be present in membranes from the facultative thermophile Bacillus coagulans grown at 55°C, compared to one in cells grown at 37°C. Cytochrome spectra and inhibitors of the respiratory chain identified them as cytochrome oxidases aa ₃ and d. Both were present in membranes from 55°C grown cells, but only cytochrome oxidase aa ₃ was found in membranes from 37°C grown cells. The presence of cytochrome d in 55°C grown cultures was found to be due to decreased oxygen tension and not to the high growth temperature. This was confirmed by (a) induction of cytochrome d at 37°C under conditions of oxygen limitation and (b) its repression at 55°C under conditions of high aeration and its subsequent induction on lowering the dissolved oxygen concentration in chemostat cultures. Two cytochromes b (_max 558 and _max 562) were present in both 37°C and 55°C grown cells. Results from the inhibition of substrate oxidation by membranes suggested different pathways of electron transport by the respiratory chain.     

Temperature regulation of oleate desaturase in sunflower (Helianthus annuus L.) seeds

The effect of temperature on oleate desaturation in developing sunflower (Helianthus annuus L.) seeds has been examined. When seeds from plants grown at low (20/10° C, day/night) temperature were transferred for 24 h to 10° C, an increase in the linoleate/oleate ratio in phosphatidylcholine and triacylglycerol was observed, but not when transfer was to 20 or 30° C. The same effect was observed in triacylglycerol, phosphatidylcholine and phosphatidylethanolamine in the newly synthesized lipids after in-vivo incubation with [1-¹⁴C]oleate at 10° C. The microsomal oleoyl phosphatidylcholine desaturase (ODS) activity of the seeds maintained at 10 C was also enhanced. The stimulation was observed after only 3 h in plants grown at high temperature (30/20° C). This effect was inhibited by cycloheximide, implying that the low-temperature stimulation of the ODS activity was caused by the synthesis of new enzyme. As a consequence, seeds from plants grown at low temperature had higher ODS activities and linoleate contents than those grown at high temperature. The microsomal ODS activity of seeds from plants grown at low temperature was dependent on incubation temperature and showed a maximum at 20° C. By contrast, this activity was almost temperature-insensitive in seeds from plants grown at high temperature. These results could explain how temperature regulates the fatty-acid composition in sunflower-seed lipids.Abbreviations DAF days after flowering - ODS oleoyl phosphatidylcholine desaturase - PC phosphatidylcholine - PE phosphatidylethanolamine - TAG triacylglycerol - 181 oleic acid - 182 linoleic acid To whom correspondence should be addressedThanks are due to M.C. Ruiz for skillful technical assistance. This work was supported by a grant from Junta de Andalucia, Spain.     

Phase transitions in plant cuticles

The effect of temperature on wet plant cuticles has been investigated with the following techniques: Calorimetry, densitometry, spin-label electron-spin-resonance-(ESR)-spectroscopy, photo bleaching, and light and electron microscopy. At low temperatures cuticles ofCitrus aurantium L. andHedera helix show, at 16.3°C, a sharp transition (T0.5°C) with a latent heat of 4.7±0.5 J g^-1-cuticle. Below transition: The main orientation of the polymer matrix is parallel to the normal of the cuticle and the main orientation of the layer with soluble lipids is perpendicular to the normal. The cuticle is in a rigid state. Above transition (between 16.3°C and 38°C): Only the orientation of the polymer matrix has changed (tilted in parts). There exist several very sharp (T0.1°C) transitions (38°C, 41°C, 45°C, 49°C, ...) with a latent heat in the order of 0.4 J g^-1-cuticle. Above 38°C: The lamella of the soluble lipids is in a fluid state. Above 45°C there is a change in the molecular orientation of the soluble lipids as well as in the polymer matrix. The soluble lipids are mainly oriented parallel to the normal. The dry cuticles show no phase transition between 0°C and 200°C. At room temperature a dry/wet transition can be observed.Abbreviations ESR-spectroscopy electron-spin-resonance-spectroscopy     

Differences in polypeptide expression in thermoinhibited and germinating achenes of Tagetes minuta L.

When imbibed at their optimum germination temperature of25°C, achenes of Tagetes minutaL.germinate over a period of 48 h. At a temperature of36°C, the achenes become thermoinhibited and do notgerminate. This elevated temperature does not harm the achenes, however, and assoon as the temperature is reduced below 35°C,germination occurs, but is spread over a reduced period of only 24h. The polypeptide patterns of germinating and thermoinhibitedachenes were compared, to test whether thermoinhibition is the result ofdifferential gene expression or repression. Ten polypeptides which weredifferentially expressed in thermoinhibited achenes were observed. Thesepolypeptides were all extremely specific to thermoinhibited achenes anddeclinedrapidly when the incubation temperature was decreased, in a manner whichcorrelated with the increase in germinability of the achenes. The tightlyregulated nature of the expression of these polypeptides suggests thatthermoinhibition in T. minuta may be under positivecontrol. These proteins may therefore result in the prevention of radicleemergence at unfavourable temperatures.     

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.     

Improvement of Kefir yeast by mutation with N-methyl-N- nitrosoguanidine

A commercial culture of Kefir grew well at 30°C and 37°C with NaCl up to 4% (w/v) but at 42°C and 4% (w/v) NaCl it grew very slowly. These conditions were selected for the mutation with N-methyl-N-nitrosoguanidine in order to improve cell yield at high temperatures and salt concentration. The mutated cells had higher biomass and growth rate compared with the original culture. The improvement of Kefir at high temperatures and salt concentrations offers advantages for uses in single cell protein production and alcohol production from whey containing high salt.