‘Safe sites’ for the seed germination ofRhus javanica: A characterization by responses to temperature and light

Germination responses ofRhus javanica L. seeds to temperature and light were investigated with special reference to their gap-detecting mechanisms in germination, i.e., responses to elevated and/or fluctuating temperatures and sensitivity to leaf-canopy transmitted light. The seeds, which have water-impermeable coats to prevent imbibition, were shown to become permeable and germinable after exposure to higher temperatures of 48–74°C for a brief period depending on the temperature. Once the coat impermeability had been removed by such heat treatment, the seeds became readily germinable over a wide range of temperature and light conditions. The lower and higher temperature limits for germination were around 8° and 36°C, respectively, with an optimal temperature of around 25°C. Simple linear relationships were observed between the temperature and germination rates, i.e., the reciprocals of the time taken by the seed subpopulations to show 10–70% germination in the sub-optimal temperature range, where the required ‘thermal time’ for germination was 2300–3600 Kh. The presence or absence of light or a simulated ‘canopy light’ had little effect on the germination of this species. It was concluded that the seeds ofR. javanica are furnished with a gap-detecting mechanism in the form of a heat requirement for the breakage of water-impermeable seed dormancy, which may be fulfilled by either daytime elevation of the surface temperature of exposed soil, or more effectively by fire.     

Storage Behaviour of Salix alba and Salix matsudana Seeds

Effects of dehydration, storage temperature and humidificationon germination of Salix alba andS. matsudana seeds were studied.Newly released seeds showed 100% germination before and afterdehydration to 11–12% moisture content. Germination ofthe high vigour lot (100% initial normal germination) was notaffected by dehydration to 6.7% moisture content but germinationdecreased with further dehydration to 4.3%. The lower vigourlot (75% initial normal germination) was more susceptible todehydration and germination decreased following dehydrationto 6.7% moisture content. Dry seeds of both species survivedimmersion in liquid nitrogen without loss of viability. Thegermination of seeds stored with 9% moisture content decreasedto 35–40% in 5 months at -20°C or in 2 months at 5°C.However, at 25°C seeds entirely lost viability within 2weeks. Seeds showed improved performance when stored at -70°C> - 20°C > 5°C > 25°C and tolerated dehydrationto a moisture content in equilibrium with 15% relative humidity.Results suggest that they are orthodox in storage behaviouralthough they are short-lived. Humidification treatment of lowvigour seed lots resulted in a remarkable increase in germinationpercentage. Copyright 2000 Annals of Botany Company Salix alba, Salix matsudana, willow, seed storage behaviour, dehydration, humidification, cryopreservation     

Photoinhibition of Seed Germination in Mediterranean Maritime Plants

Photoinhibition of seed germination was shown for Allium staticiforme,Brassica tournefortii, Cakile maritima and Otanthus maritimus,all plant species inhabiting sandy coasts of the MediterraneanSea. Germination of A. staticiforme was found to be typicallyMediterranean in regard to its temperature range (>0–20°C), while B. tournefortii germinated optimally at intermediatetemperatures (15–25 °C). Light sensitivity was morepronounced in the latter species and 50 % inhibition of seedgermination was obtained with photon flux densities of approximately0.1 and 0.015 mol m^–2 d^–1 in A. staticiforme andB. tournefortii, respectively. However, the slopes of the regressionlines of germination plotted against the logarithm of whitelight flux density are similar in both species. From monthlyexperiments performed under fluctuating conditions of temperatureand light, simulating the elimate of Athens throughout the year,optimal germination response peaked in ‘winter’conditions for A. staticiforme, while a bimodal pattern wasobserved for B. tournefortii (maxima at the warm ends of therainy season). In contrast to the other plants, a fifth speciestested, Crithmum maritimum, showed an absolute light requirement,probably related to the rocky habitats of the species; the optimumgermination period coincided with the rainy season. Pot experimentswith seeds of the five species buried at various depths confirmedthat maximum emergence is favoured by shallow depths (0.5–1cm). It is concluded that seed germination in maritime plantsof the Mediterranean rim is mediated by a photoinhibition mechanismwhich can be considered an adaptation strategy against surfaceseedling establishment at the harsh, sandy or shingle, sea coasthabitats. Allium staticiforme, Brassica tournefortii, Cakile maritima, sea rocket, Crithmum maritimum, rock samphire, Otanthus maritimus, ton-weed, seed germination, light, photoinhibition, phytochrome, seedling emergence     

Seed Germination and Seedling Development in Cyclamen persicum

Cyclamen persicum Mill, seeds germinate in a narrow range oftemperature and germination is strongly inhibited by continuousirradiation with white light. The thermal optimum is approx.15 °C in both darkness and light. Seed germination is alsovery sensitive to oxygen deprivation and this sensitivity ismore pronounced at 20 °C than at the optimum 15 °C.Very immature seeds cannot germinate at any temperature, butgerminability increases during seed maturation Seedling development is unusual since seed reserves are usedimmediately for tuber formation. Tuberization is optimal at15–20 °C in light and in darkness. Supra-optimal temperatures(25–30 °C) or hypoxia inhibit tuber formation andlead to very elongated tubers These results allow the producers to improve the productionof homogeneous populations of cyclamen seedlings Wheat seeds, Triticum aestwum L., acetylcholinesterase, electrophoresis, germination, assay     

Effects of Temperature and Water Potential on Germination, Radicle Elongation and Emergence of Mungbean

Controlled environment experiments were performed to determinethe effects of temperature and water potential on germination,radicle elongation and emergence of mungbean (Vigna radiata(L.) Wilczek cv. IPB-M79-17-79). The effects of a range of constant temperatures (15–45°C) and water potentials (0 to –2.2 MPa) on germinationand radicle elongation rates were studied using an osmoticumtechnique, in which seeds were held against a semi-permeablemembrane sac containing a polyethylene glycol solution. Linearrelationships were established between median germination time(Gt₅₀) and water potential at different temperatures, and betweenreciprocal Gt₅₀ (germination rate) and temperature at differentwater potentials. Germination occurred at potentials as lowas –2.2 MPa at favourable temperatures (30–40 °C),but was fastest at 40 °C when water was not limiting, withan estimated base temperature (T_b) of about 10 °C. Subsequentradicle elongation, however, was restricted to a slightly narrowertemperature range and was fastest at 35 °C. The conceptof thermal time was used to develop an equation to model thecombined effects of water potential and temperature on germination.Predictions made using this model were compared with the actualgermination obtained in a related series of experiments in columnsof soil. Some differences observed suggested the additionalimportance of the seed/soil/water contact zone in influencingseed germination in soil. Seedling emergence appeared to reflectfurther the radicle elongation results by occurring within anarrower range of temperatures and water potentials than germination.Emergence had an estimated T_b of 12.6 °C and was fastestat 35 °C. A soil matric potential of not less than about–0.5 MPa at sowing was required to obtain 50% or moreseedling emergence. Key words: Germination, temperature, water potential     

Gap-detecting mechanism in the seed germination ofMallotus japonicus (Thunb.) Muell. Arg., a common pioneer tree of secondary succession in temperate Japan

Germination responses ofMallotus japonicus (Thumb). Muell. Arg. seeds to temperature revealed a gap-detecting mechanism in the seed germination of the species. Among various constant and alternating temperatures examined in the range from 12–40°C, only very limited temperature regimes were found to be favourable for seed germination, specifically, alternating temperatures between 18–32°C and 28–40°C. A single several-hour higher-temperature (32–40°C) treatment could also induce the germination of seeds which had been imbibed for several days at a constant temperature in the range of 20–26°C, suggesting that there is a process requiring higher temperature among the overal germination processes. Seeds located at or near the surface of denuded soil would have a good chance of experiencing such a temperature change when several rainy days are followed by fine weather, while seeds beneath close vegetation would not. On the other hand, the pressence or absence of light or a simulated ‘canopy ligh’ had little effect on the germination. Therefore, it was concluded that the seeds ofM. japonicus have a ‘gapdetecting mechanism’ in the form of a higher-temperature requirement of a certain process involved in the overall germination processes.     

The Effects of Temperature and Osmotic Moisture Stress on the Germination of Lotus corniculatus

Germination was delayed by temperatures lower than 15 ?C, anddelayed and reduced by temperatures of 30 ?C or higher. Germinationwas buffered against temperature and osmotic moisture stressesin the region 10–25 ?C and 0 to –4 bars. But athigher temperatures or osmotic moisture stresses, both factorsdelayed and reduced germination, and one factor intensifiedthe effect of the other. The germination of the Empire varietywas reduced less than that of Viking or Mansfield by temperaturesof 30 ?C or higher. The germination of small seeds was reducedmore than that of large seeds by temperatures of 25 ?C or higher.There were no significant differences between varieties or seedsizes in their over-all responses to moisture stress, but thetimes of some of the responses differed according to the varietyor seed size.     

Quantal Response of Fruit and Seed Germination Rate in Quercus robur L. and Castanea sativa Mill, to Constant Temperatures and Photon Dose

A linear relationship between constant temperatures in the sub-optimaltemperature range and germination rate is shown in both Quercusrobur L and Castanea sativa Mill germinated under nominal darkconditions. The mean base temperature was interpolated for Qrobur as 0 8 ? or 2-4 ?, depending on seed lot provenance, andfor C. sativa as 1 -4? The optimum temperature for germinationin Q. robur was about 20? compared with around 28 ? in C. sativaOver the sub-optimal temperature range the distribution of thermaltimes was log-normal for each population studied their spreadvarying both between Q robur seed lots and between species However,in C. sativa germinated close to the mean base temperature,the distribution in thermal times was reduced Thermal timesto germination were decreased in Q. robur and C sativa by approximately0 3 and 0-5 log-units, respectively, when the pericarp was removed,i.e in the seeds, but the sensitivity of the response remainedrelatively unaltered In both species the germination rate was the same when nominaldark or safe green light conditions were employed during thegermination test. However, at 21 ? Q robur exhibited the highirradiance reaction (HIR) at photon doses above 30mmol m^–2d^–1. HIR first affected the germination rate by an inhibitionof radicle extension The sensitivity of the response to thermaltime was reduced as photon dose increased. This photo-inhibitionwas exacerbated at supra-optimal temperatures. In contrast,C. sativa germination rate at 26 ? was little influenced bylight at a photon dose of 752 mmol m^–2 d^–1 Key words: Seed germination rate, temperature, thermal time, light, photon dose     

Effect of Water Potential and Temperature on the Germination of Four Species of African Savanna Trees

The germination responses of seeds from the African tree speciesColophospermum mopane, Combretum apiculatum, Acacia tortilisand Acacia karroo under varying regimes of temperature and waterstress (induced by incubation in PEG 8000) are reported Withthe exception of Combretum (at –0.14 and –0.29 MPa)and Colophospermum (at –0.29 MPa), incubation in PEG decreasedthe maximum achieved germination percentage (90–100% forall species), but did not extend the germination lag (exceptin Combretum) or affect the time required to reach maximum germinationCombretum and Colophospermum were found to germinate under thewidest range of temperatures and water potentials, for example,as strongly negative as –1.0 MPa at 20 and 30 °C,respectively These seeds also showed greater or equivalent hypocotylelongation in PEG solutions creating potentials of –0.14,–0.29 or –0.51 MPa when compared with seeds germinatedin water, indicating an additional stress adaptation Acaciaspecies showed progressive reduction in germination rates andradicle elongation in response to decreasing water potentialExperiments giving pre-imbibition treatments in water priorto transfer to PEG solutions showed that both Acacia speciesgerminated at approximately 90% if given such pre-treatmentand less than 10% if transferred directly to PEG It is concludedthat the most stress-adapted species studied are Colophospermummopane and Combretum apiculatum, a finding generally correlatedwith the growth habit of these trees Colophospermum mopane, Combretum apiculatum, Acacia tortilis, Acacia karroo, germination, water stress, Zimbabwe     

A Scanning Electron Microscopy Study of the Seed and Post-seminal Development in Angelonia salicariifolia Bonpl. (Scrophulariaceae)

The external morphology of seeds and post-germination developmentalstages of Angelonia salicariifolia Bonpl. (Scrophulariaceae)were investigated using scanning electron microscopy. Some structuralfeatures of the seed exotesta and seedling in Angelonia arepresented for the first time and are of potential taxonomicvalue for this neotropical genus. The seeds are very small (0.9–1.7mm long and 0.5–0.9 mm wide), ovate, with a reticulate-crestedexotesta, reticules arranged uniformly in longitudinal rows,with a high density of microcilia-like projections on the cellwall of the reticule base and on the edge of the crests. Thehilum is located beside the micropyle at the narrow end of theseed. Germination is epigeal. During germination the radicledevelops, followed by elongation of the hypocotyl and primaryroot. At this stage dense root hairs develop on the lower partof the hypocotyl. The apical bud—located between the cotyledons—beginsto develop after the cotyledons have unfolded. The cotyledonsare equal in size, sessile and ovate. The seedlings have twotypes of trichomes, one characteristic of the cotyledons andfirst pair of leaves (glandular, sessile, four-celled head withquadrangular shape) and the other characteristic of the hypocotyland epicotyl (stalked, erect, elongate and three-celled withdome-shaped unicellular head). Copyright 2001 Annals of BotanyCompany Seed morphology, exotesta-ornamentation, micromorphology, post-seminal stages, seedling, trichomes, SEM, Angelonia salicariifolia Bonpl., Scrophulariaceae     

The Effects of Priming and Ageing on Seed Vigour in Tomato

A comparison was made of the effects of seed priming or ageingtreatments on the performance of tomato (Lycopersicon esculentumMill. cv. UC204C) seeds according to a number of indices ofseed vigour. A single lot of tomato seeds was primed in 120mol m^–3 K2HPO4 + 150 mol m^–3 KNO₃ for 5 d at 20?C, or aged at 13% moisture content (dry weight basis) and 50?C for 6 d. Germination percentage (>98%) was unaffectedby priming and reduced to 85% by ageing. X-ray photographs andlongitudinal sections revealed the formation of free space surroundingthe embryo in dry primed seeds, which was not evident in controlor aged seeds. Priming increased the rate of germination atall temperatures above the base temperature (T_b), while ageingdecreased it. T_b was unaffected by priming and only slightlyincreased by ageing. The variation in individual times to germinationwas approximately doubled in both primed and aged seed comparedto the control, based upon the slopes of probit germinationpercentage versus log thermal time curves. Root growth aftergermination tests and seedling growth in both greenhouse andfield tests were not influenced by either priming or ageing.The conductivity test was found to be unreliable as a vigourtest for tomato seeds. The results identify several indiceswhich can be used to quantify seed vigour in tomato. They alsoillustrate that seed priming can enhance seed performance accordingto some criteria, while having no effect or decreasing qualityaccording to other criteria. Seed vigour can apparently be separatedinto various components which can be independently influencedby seed enhancement treatments. Key words: Tomato, seed germination rate, seed priming, seed vigour     

Promotion by gibberellin of lettuce seed germiation as a function of presoaking period

Germination of lettuce seeds (Lactuca sativa L. cv. Grand Rapids)was examined in the presence of various doses (10^–5.0–10^–3.0M) of gibberellic acid applied at various times (hour 0–8)of soaking. Germination promotion by gibberellic acid was greateras the dose of gibberellic acid was increased and attenuatedwith the length of the presoaking period. As an exception, ca.95% germination was always evoked by the largest dose (10^–3M) of gibberellic acid given at any time of soaking. The dose-responsecurve obtained for each presoaking period had a distinct sigmoidalprofile. Synergistic and photoreversible promotion by red light of thegibberellin-induced germination was also investigated. Far-redlight pulse given 6 hr after the red pulse was still effectivein removing the red light action. Application of enzyme kineticsto the gibberellin action and also to the synergism betweengibberellin and red light was suggested. ¹National Institute for Basic Biology, Myodaiji, Okazaki 444,Japan. (Received December 18, 1979; )     

Effects of Moisture Content on Germination of Seeds of Hancornia speciosa Gom. (Apocynaceae)

Seeds of Hancornia speciosa germinated best at a temperatureof 20–30 °C. The viability of the seeds during storagewas short and the best storage conditions for viability entailedkeeping the seeds in polyethylene bags. Seed viability was maintainedonly when the seeds were stored at a moisture content above30%; storage conditions which allowed dehydration resulted ina rapid loss of viability (the seeds showed recalcitrant behaviour). Low temperature during storage did not improve longevity. Arelationship between germination and moisture content was established,but when the moisture content fell below 25% there was a drasticreduction of germination. After 9 weeks of storage, even athigh moisture content, seeds lost viability. Loss of seed viability during seed dehydration was associatedwith increased leakage of electrolytes and organic solutes,and reduced tetrazolium staining during subsequent imbibition. Hancornia speciosa, germination, recalcitrant seeds, storage, moisture     

Dormancy in seeds of Phellodendron wilsonii is Mediated in Part by Abscisic Acid

A regime of temperatures alternating between 35°C (8 h)and 10°C (16 h) proved to be a specific method for breakingdormancy of seeds of Phellodendron wilsonii. The relationshipbetween the germination capacity after about 13 days of incubationunder this regime and the logarithm of the amount of ABA inthe seed before incubation yielded a high correlation coefficient.The final germination capacity under this regime was poorlycorrelated with the amount of ABA in seeds before incubation.These observations suggest that at least part of the germinationbehavior is controlled by the amount of abscisic acid in theseed. Stratification decreased the amount of ABA but resultedin only 8–25% germination relative to other dry storedseeds failed to germinate under suboptimal conditions, namely,incubation at 22°C with 12 h light. This phenomenon indicatesthat some unknown mechanism is the major factor that controlsthe dormant state, and the effect of this mechanism are overcomeonly by the specific alternating-temperature regime. Fluctuationsin temperature failed, however, to reduce the ABA content ofseeds during the incubation period. We conclude that some unknownfactor contributes to the dormancy of seeds of P. wilsonii althoughthe level of ABA definitely plays a minor role in the maintenanceof seed dormancy. (Received January 25, 1993; Accepted November 25, 1993)     

Responses of Vegetable Seeds to Controlled Hydration

Leek, onion and carrot seeds were imbibed in water and in solutionsof polyethylene glycol (PEG) 6000 over the range –0.5to –4.0 MPa osmotic potential, for periods up to 28 dat 15 C. Seeds started to germinate after 7 and 14 d at –0.5MPa and –1.0 MPa PEG, respectively, but in the lattercase, germination did not exceed 5%. No germination occurredin solutions of lower (more negative) osmotic potential. Seedmoisture content increased with osmotic potential in all threespecies and the relationships were unaffected by the durationof imbibition in solutions of –1.0 to –4.0 MPa,though leek seeds had higher moisture contents than the otherspecies for any given osmotic potential. Linear relationships between response to priming (differencebetween mean germination times of primed and untreated seeds)and seed moisture content were obtained for each species, positiveresponses being obtained above 30–35% seed moisture contentwith optima at 46, 44.5 and 44% seed moisture contents in leek,onion and carrot, respectively. Priming had no effect on embryovolume or cell number per embryo in leek and onion. Carrot embryovolume increased by 43% and cell number per embryo doubled inprimed compared with untreated seeds, whereas seeds imbibedin water showed only a slight increase in cell number per embryoat the stage when radicles were beginning to penetrate the seedcoat. Allium cepa L. cv. Rijnsburger Robusta, onion, Allium porrum L. cv. Winterreuzen, leek, Daucus carota L. cv. Nantaise, carrot, germination, priming, polyethylene glycol, seed moisture, cell number, embryo volume     

Light Actions in the Germination of Cocklebur Seeds: I. DIFFERENCES IN THE LIGHT RESPONSES OF THE UPPER AND LOWER SEEDS

Germination responses to light were studied in the upper andlower seeds of cocklebur (Xanthium pennsylvanicum Wallr.). Thelower seed was dark-germinating and negatively photoblastic;the upper one had a red-light (R) requirement and was positivelyphotoblastic. Germination of the lower seeds was inhibited bya prolonged single irradiation with R, blue (B) or far-red (FR)light applied during imbibition. The maximal inhibitory effectof a single irradiation occurred 9 h and 13 h after the startof soaking at 33 °C and 23 °C, respectively. However,the inhibitory effect of R differed from that of B and FR, byonly delaying germination. A single exposure to B or FR lightcould be replaced by intermittent B or FR irradiation, and theireffects were repeatedly reversible by the following R irradiation.If the upper seeds were not exposed to R during imbibition,they failed to germinate even at 33 °C which was optimalfor germination, and the promotive effect of R increased withdelay of its application time. The photoperceptive locus incocklebur seeds was the axial tissue for all B, R and FR. Lightreceived by the cotyledonary tissue had little effect. Germinationdimorphism in response to light is discussed with respect tothe phytochrome content and the ageing of axial tissues. Key words: Blue light, Dimorphism, Far red light, Germination, Red light, Xanthium seed     

An Intermediate Category of Seed Storage Behaviour?II. EFFECTS OF PROVENANCE, IMMATURITY, AND IMBIBITION ON DESICCATION-TOLERANCE IN COFFEE

Seeds of four cultivars of arabica coffee (Coffea arabica L.)received from three continents survived desiccation to between7-2% and 11-3% moisture content (wet basis), i.e. to seed waterpotentials of –90 MPa to –150 MPa, but further desiccationreduced germination (criterion, normal seedling development)in all seed lots. Only a few individuals from four of the lotsgerminated after being dried to 4–5% moisture content.Differences in desiccation sensitivity were apparent among lotswithin each cultivar. Desiccation sensitivity in these lotswas similar to that observed in seeds of orthodox species whichhave begun to germinate. Seeds extracted from fruits of intermediatematurity (yellow) were able to tolerate greater desiccationthan those from either ripe (red) or immature (green) fruits.Imbibed storage increased desiccation sensitivity. The resultsare compatible with the view that arabica coffee seeds are unableto tolerate extreme desiccation because germination has beeninitiated before harvest.     

Genotypic Differences in the Temperature Responses of Tropical Crops: I. GERMINATION CHARACTERISTICS OF GROUNDNUT (ARACHIS HYPOGAEA L.) AND PEARL MILLET (PENNISETUM TYPHOIDES S. & H.)

Mohamed, H. A., Clark, J. A. and Ong, C. K. 1988. Genotypicdifferences in the temperature responses of tropical crops.I. Germination characteristics of groundnut (Arachis hypogaeaL.) and pearl millet (Pennisetum typhoides S. & H.).—J.exp. Bot. 39: 1121–1128. The germination at constant temperature of several genotypesof groundnut and pearl millet was investigated between 0?C and50?C on a thermal gradient plate. Large differences in bothgermination rate and percentage germination were observed inboth species. Base temperatures vary from 8–11.5?C and 8–13.5?Cin groundnut and millet, respectively and optimum temperaturesfrom about 29–36.5?C in both. Maximum temperatures forgermination ranged from 41–47?C. The results are discussedin terms of adaptation to high soil temperature and crop establishmentin the semi-arid tropics. Key words: Temperature, germination, millet, groundnut     

Water as a Storage Medium for Elaeis guineensis (pisifera) Seeds under Aseptic Conditions

Over 50 per cent germination has been obtained from Elaeis guineensisform pisifera seeds stored in unaerated sterile distilled waterfor 6 months. The moisture levels of the seeds and excised embryoswere of the same order (20–30 and 60–70 per cent,respectively) as those of fully imbibed fresh seeds. The implicationsof an apparent lower oxygen requirement by seeds stored underwater as against germinating seeds are discussed in the contextof the successful storage. Elaeis guineensis, pisifera, germination, seed dormancy, embryo     

A Model for Germination Responses to Alternating Temperatures

When seeds of Chenopodium album are imbibed in 0–01 Mpotassium nitrate solution in the light at constant temperature,percentage germination increases to an optimum at 24 °C,above which it decreases. These relationships are linear ifpercentage germination values are transformed to normal deviates.At supra-optimal temperatures, alternating temperatures havelittle or no effect on this basic relationship. However, atsub-optimal temperatures normal deviate germination increaseslinearly with amplitude at constant mean temperature and theincreases are relatively greater at lower mean temperatures.The effect of amplitude is also greater when more time is spentat the wanner temperature in the diurnal cycle. Seeds of Panicummaximum show very similar responses except that the effect ofamplitude is greater when a shorter time is spent at the warmertemperature in the diurnal cycle. These observations form thebasis of a quantitative model which, at sub- and supra-optimaltemperatures, respectively, accounts for 90 and 75% of the variationin germination of C. album seeds subject to a very wide rangeof thermal environments on a two-dimensional temperature gradientplate. The data presented for P. maximum are less comprehensive,but again the model accounts for 80% of the variation. The relevanceof the same model to two unrelated species from different climatesand of different ecological behaviour suggests that it may begenerally useful in determining optimum temperatures for seedviability testing regimes and ultimately for predicting fieldbehaviour These results allow the producers to improve the productionof homogeneous populations of cyclamen seedlings Chenopodium album L., Panicum maximum Jacq., seed dormancy, germination, alternating temperatures, temperature gradient plate