Shoot Apical Meristems of Picea sitchensis Seedlings Accelerate in Growth Following Bud-set

First-year seedlings of Picea sitchensis were induced to setbuds by transferring them from long days (17 h) to short days(10 h at 20 °C, 12 h night at 12 °C). After 20 shortdays, the seedlings stopped growing in height, the shoot apicaldomes began enlarging and the transverse growth rates of theshoot apical dome tissues increased. For about 25 days thereafterprimordia were produced faster within the developing buds thanat the apices of seedlings which had been kept in long daysto prevent bud-set. Accelerated primordia production after bud-setenables the largest possible buds to be formed during the frost-freeperiod before winter. Picea sitchensis, apical meristem, bud-set     

Leaf Extension in Zea mays: II. LEAF EXTENSION IN RESPONSE TO INDEPENDENT VARIATION OF THE TEMPERATURE OF THE APICAL MERISTEM, OF THE AIR AROUND THE LEAVES, AND OF THE ROOT-ZONE

The temperatures of the roots, the apical meristem, and theshoots of Zea mays plants were varied independently of eachother and the rates of leaf extension were measured. When thetemperature of the apical meristem and region of cell expansionat the base of the leaf was kept at 25 °C, changes of leafextension in response to changes of root and shoot temperatureswere less pronounced. When the temperature of the meristematicregion was changed by increments of 5 or 10 °C from 0 to40 °C, and the root and shoot temperatures were kept at25 °C, rapid changes in leaf extension occurred. It was concluded that the rates of leaf extension were controlledat root-zone temperatures of 5 to 35 °C by heating or coolingof the meristematic region. Changes in rates of leaf extensionin response to changes in air temperature were attributed todirect effects on the temperature of the meristematic regionand on the physiology of the leaf.     

Micronuclei and Radiosensitiviy in the Root Meristem of Vicia faba

Percentages of cells with micronuclei in four regions of theroot meristems of Vicia faba are used as measures of sensitivityto acute X-irradiation. There are two peaks in these percentages,occurring at about four and eight days after 360 rads and twoand six days after 180 rads. Two peaks exist, probably becausethe radiation delays cells that were in G₁ much more than cellsthat were in G₂ in reaching the first post-irradiation mitosisand consequently in displaying micronuclei in the followinginterphase. The relative heights of the two peaks thereforereflect the relative numbers of cells in G₁ and G₂ as well asthe relative sensitivity of the two phases to chromosomal damage. The cells of the quiescent centre are injured least by the radiationas they are mostly held at Gt. The meristem thus obeys the lawof Bergonié and Tribondeau, but differs from that ofZea in that the meristematic cells of the cap initials and steleimmediately adjacent to the quiescent centre resemble the quiescentcentre much more closely than the stele 250 µ away inthe numbers of micronuclei produced. This is consistent withthe differences already known between the two species concerningrates of division in the different regions of the meristem andthe behaviour of the meristem after severe radiation injury.     

Effects of Low Temperature on Growth and Nutrient Accumulation in Rye (Secale cereale) and Wheat (Triticum aestivum)

Rye (Secale cereale cv. Rheidol) and wheat (Triticum aestivumcv. Mardler) were grown at shoot/root temperatures of 20/20°C (warm grown, WG plants), 8/8 °C (cold grown, CG plants)and 20/8 °C (differential grown, DG plants). Plants fromcontrasting growth temperature regimes were standardized andcompared using a developmental timescale based on accumulatedthermal time (°C d) at the shoot meristem. Accumulationof dry matter, nitrogen and potassium were exponential overthe time period studied (150–550 °C d). In rye, therates of plant dry matter and f. wt accumulation were linearlyrelated to the temperature of the shoot meristem. However, inwheat, although the rates of plant dry matter and f. wt accumulationwere temperature dependent, the linear relationship with shootmeristem temperature was weaker than in rye. The shoot/rootratio of rye was stable irrespective of growth temperature treatment,but the shoot/root ratio of wheat varied with growth temperaturetreatment. The shoot/root ratio of DG wheat was 50% greaterthan WG wheat. In both cereals, nutrient concentrations anddry matter content tended to be greater in organs exposed directlyto low temperatures. The mean specific absorption rates of nutrientswere calculated for the whole period studied for each species/temperaturecombination and were positively correlated with both plant shoot/rootratio and relative growth rate. The data suggest that nutrientuptake rates were influenced primarily by plant demand, withno indication of specific nutrient limitations at low temperatures. Nutrient accumulation, relative growth rate (RGR), rye, Secale cereale cv. Rheidol, temperature, thermal time, Triticum aestivum cv. Mardler, wheat     

The Quiescent Centre and Rates of Mitosis in the Root Meristem of Allium sativum

The root apices of Allium sativum have been examined by continuous-and pulse-labelling with tritiated thymidine and by colchicinetreatment to measure the time parameters of the mitotic cyclein various parts of the meristem. There is a quiescent centre of 30–50 cells whose averagerate of mitosis is low because the G₁ period is extended toabout 140 h compared with about 4 h in the othe regions of themeristem. The stele just above the quiescent centre and at 200microns above it and the cap initials just below the quiescentcentre are very similar in their mitotic cycles, the total lengthsof which are about 30 h of which nearly half is taken up byDNA synthesis. Allium thus differs from Zea in having root capinitials whose mitotic cycle is not telescoped by the eliminationof the G₁ phase. These facts are discussed in relation to theradiosensitivity of the meristem.     

Feeding of freshwater filter-feeders at very low food concentrations: Poor evidence for "threshold feeding" and "optimal foraging" in Daphnia longispina and Eudiaptomus gracilis

The feeding behaviour of two freshwater zooplankton species,the calanoid copepod Eudiaptomus gracilis and the cladoceranDaphnia longispina, at extremely low food concentrations hasbeen studied in order to test whether the animals react as predictedby the models of optimal foraging. Three different sized algae were offered in concentrations downto 1µg C/litre at 7°C during winter and 19°C duringsummer. Ingestion rates were measured by use of a flow-through¹⁴C-technique In contrast to the findings of several authors working withmarine copepods a threshold concentration where the animalsceased filtering could be detected only in one experiment byregression analysis (Daphnia, 19°C, food: Stichococcus).A depression of the filtering rate at low concentrations wasobserved only in Daphnia at 19°C with Stichococcus and Scenedesmusas food. It is supposed that in this case the reduction of thefiltering rate is not a direct response of the filtrator, butis caused by exhaustion due to the experimental conditions. As freshwater zooplankton is very rarely exposed to food abundancesas low as in the marine environment, there seems to be no selectionpressure favouring the evolution of an "optimal forager" behaviour. ⁽¹⁾Supported by Deutsche Forschungsgemeinschaft     

Temperature-dependent consumption and gut-residence time in the opossum shrimp Mysis relicta

Maximum daily consumption was estimated for Mysis relicta fedad libitum rations of Daphnia pulex at 4,10,15 and 18°C.Gut-residence time was also evaluated for M.relicta fed clado-ceranprey at 4, 10 and 157deg;C. Mean daily consumption (g dry weightof Daphnia g^–1 dry weight of Mysis day^–1) rangedfrom 6% at 4%C to 12% at 10°. At 18°C, Mysis feedingrate declined to 9% day1. Mean, weight-adjusted consumptionrates exhibited a ‘dome-shaped’ response in relationto water temperature. Consumption rate was highest at 10°Cand lowest at 4°C. Estimated Q₁₀ was more sensitive from4 to 10°C (Q₁₀= 3) than from 10 to 15°C (Q₁₀=1.2). Gut-residencetime for Mysis was inversely related to water temperature, implyingthat evacuation rate increases linearly with water temperature.Feeding and gut-evacuation rates become disassociated at watertemperatures >10°C. As water temperature increased above1°C, relative evacuation rate increased, whereas feedingrate declined. It is postulated that at higher water temperatures,disassociated feeding and gut-evacuation rates reduce the scopefor growth of vertically migrating Mysis and impose a physiologicalconstraint that isolates Mysis from warm, epilimnetic waterduring thermal stratification. ¹Present address: Center for Aquatic Ecology, Illinois NaturalHistory Survey, Sam Parr Biological Station, 6401 Meacham Road,Kinmundy, IL 62854, USA     

Effect of Root and Shoot Meristem Temperature on Shoot to Root Dry Matter Partitioning and the Internal Concentrations of Nitrogen and Carbohydrates in Maize and Wheat

Maize (Zea mays L.) and spring wheat (Triticum aestivum L.)were grown in nutrient solution at uniformly high air temperature(20 °C), but different root zone temperatures (RZT 20, 16,12 °C). To manipulate the ratio of shoot activity to rootactivity, the plants were grown with their shoot base includingthe apical meristem either above (i.e. at 20 °C) or withinthe nutrient solution (i.e. at 20, 16 or 12 °C). In wheat, the ratio of shoot:root dry matter partitioning decreasedat low RZT, whereas the opposite was true for maize. In bothspecies, dry matter partitioning to the shoot was one-sidedlyincreased when the shoot base temperature, and thus shoot activity,were increased at low RZT. The concentrations of non-structuralcarbohydrates (NSC) in the shoots and roots were higher at lowin comparison to high RZT in both species, irrespective of theshoot base temperature. The concentrations of nitrogen (N) inthe shoot and root fresh matter also increased at low RZT withthe exception of maize grown at 12 °C RZT and 20 °Cshoot base temperature. The ratio of NSC:N was increased inboth species at low RZT. However this ratio was negatively correlatedwith the ratio of shoot:root dry matter partitioning in wheat,but positively correlated in maize. It is suggested that dry matter partitioning between shoot androots at low RZT is not causally related to the internal nitrogenor carbohydrate status of the plants. Furthermore, balancedactivity between shoot and roots is maintained by adaptationsin specific shoot and root activity, rather than by an alteredratio of biomass allocation between shoot and roots.Copyright1994, 1999 Academic Press Wheat, Triticum aestivum, maize, Zea mays, root temperature, shoot meristem temperature, biomass allocation, shoot:root ratio, carbohydrate status, nitrogen status, functional equilibrium     

Effect of Temperature on Interception and Conversion of Solar Radiation by Stands of Groundnut

Stands of groundnut were grown in controlled environment glasshousesat air temperatures of 19, 22, 25, 28, and 31°C. Leaf areaindex (L) increased with rise of temperature, and after 85 dwas about 10-fold larger at 31°C than 19°C. Over mostof the range of temperature, both L and fractional interceptionof solar radiation (f) were functions of thermal time accumulatedfrom sowing (above a base of 10°C). In this respect, theywere tightly coupled to developmental rate at the main apex.In one experiment, only 38% of seeds emerged at 22°C and21% at 19°C, compared with more than 70% at 25°C and31°C, but the low population density was compensated byfaster leaf expansion by each plant (at 22°C only) and agreater fraction of solar radiation intercepted by unit leafarea. The amount of solar radiation intercepted by stands increasedwith rise in temperature, but the greatest differences betweentreatments occurred before the canopies achieved complete groundcover (i.e.f>0.9) and the relative effect of a rise in temperaturediminished the longer the duration of growth. The dry matterproduced for unit solar radiation intercepted was not stronglyaffected by temperature between 22°C and 31°C, wherethe mean was 2.1 g MJ^–1; the value at 19°C was uncertainsince the stands were sparse throughout the experiment. After85 d, the stand at 31°C had produced eight times the drymatter of that at 19°C—a difference caused mainlyby the effect of temperature on the rates of development andexpansion. Key words: Dry matter production, groundnut, radiation interception, temperature, thermal time, roses     

Plant Regeneration Through Meristem Culture from Vegetative Buds of Mulberry (Morus bombycis Koidz.) Stored in Liquid Nitrogen

Intact vegetative buds of mulberry (Morus bombycis) attachedto shoot segments were prefrozen, stored in liquid nitrogen,thawed, and the meristems excised for culture on Murashige andSkoog's medium supplemented with 1 mg 1^–1 BA to regenerateplants. Either prefreezing at –10 °C or –20°C along with rapid thawing at 37 °C or prefreezingat –20 °C or –30 °C along with slow thawingat 0 °C was a suitable condition for high percentages ofsurvival and shoot regeneration. Potted mulberry plants couldbe finally obtained from the cryopreserved material. The systemusing intact buds as the material for cryopreservation is quitesimple when compared to conventional systems using isolatedmeristems together with cryoprotectants. Morus bombycis Koidz., mulberry, cryopreservation, meristem culture, plant regeneration     

Effect of temperature on growth and ingestion rates of Favella sp

This study describes the effect of temperature on the growthand ingestion rates of the tintinnid, Favella sp. cultured withthe dinoflagellate Heterocapsa triquetra. In vivo fluorescencewas used to monitor the change in density of the H. triquetrapopulation over 4- to 5-day periods in control tubes containingonly algae, and in experimental tubes containing algae and tintinnids.A ‘switchover point’ occurred in the temperaturedependency of the growth rate such that below 11.4°C, H.triquetra grew more quickly than Favella sp. and above thistemperature the situation was reversed. Ingestion rates of Favellaon H. triquetra were found to be temperature dependent in anonlinear fashion. The rate doubled (from 2.5 to 5.3 cells animal^–1h^–1) between 11.4 and 16.4°C whereas there was nochange in ingestion rates between 8.0 and 11.4°C, or between16.4 and 21.1°C.     

Feeding and growth rates of the doliolid, Dolioletta gegenbauri Uljanin (Tunicata, Thaliacea)

The goal of this research is to enhance our knowledge of thecontributions of doliolids to the planktonic community as consumersand secondary producers. The objectives are to quantify feedingand growth rates of Dolioletta gegenbauri gonozooids at fourfood concentrations and four temperatures in order to determinetheir impact as grazers throughout the water column. Althoughdoliolids are abundant in numerous regions of the coastal ocean,and are considered to be major planktonic grazers, data on ratesof feeding and growth are scarce. Laboratory experiments wereconducted at 16.5, 20, 23.5 and 26.5°C to quantify removalof a 50:50 volumetric concentration of Thalassiosira weissflogiiand Rhodomonas sp. at four different food concentrations of20, 60, 160 and 390 µg C l^–1. Results from theseexperiments suggest that clearance rates are similar at concentrationsfrom 20 to 60 µg C l ^–1, and decrease as the foodconcentrations increase to 160 and 390 µg C l ^–1.The ingestion rates increase over a range of phytoplankton concentrationsfrom 20 to 160 µg C l ^–1, then decrease when abnormallyhigh concentrations of 390 µg C l ^–1 are offered.Clearance and ingestion rates increase as temperature increasesfrom 16.5 to 26.5°C. The exponential growth rates rangefrom k = 0.2–0.7, with the lowest rates occurring at thehighest food concentration. Growth rates increase with increasingtemperature from K = 0.1–0.3 at 16.5°C to 0.45–0.7at 26.5°C. In each case, the small- and medium-sized zooidshad higher growth rates than the larger gonozooids. These resultssuggest that doliolid feeding and growth rates are a functionof environmental food concentrations and temperatures, and implythat they can be important consumers in a changing neritic environment.     

Changes in Ultrastructure and Abscisic Acid Level, and Response to Applied Gibberellins inTaxus maireiSeeds Treated With Warm and Cold Stratification

Seeds ofTaxus maireiare known for their deep dormancy whichcan only be broken by a procedure involving warm stratificationfollowed by cold stratification. Treatments with alternatingtemperatures of 25/15 or 23/11 °C (12 h light) for 6 monthsfollowed by 5 °C for 3 months were successful in overcomingseed dormancy. After 6 months of warm stratification, cytologicalchanges observed included: enlargement of the embryo; a decreasein the number of lipid bodies; appearance of ER; and increasesin mitochondria, plastids, dictyosomes, vacuoles and microbodiesin the shoot apical meristem. Cold stratification followingthe warm treatment induced cell division, and one or two distinctnucleoli in the shoot apical meristem cells were observed. Bothwarm and cold stratification reduced endogenous ABA concentrationsfrom the original 8888 pg per freshly harvested seed to 392and 536 pg, respectively. Treatment with exogenous gibberellinsafter seeds had been warm-stratified showed that GA₄and GA₇wereeffective at promoting seed germination, but GA₃was not. Theseresults suggest that the strong seed dormancy ofT. maireicouldbe caused by a high ABA content and underdevelopment of theembryos in freshly shed seeds. We conclude that warm stratificationwith alternating temperatures increases the growth of embryosby cell expansion and enlargement and decreases ABA content,but seeds still remain ungerminated. Cold stratification mayinduce the response to GAs and initiate cell division resultingin release from physiological dormancy and subsequent germinationofT. maireiseeds.Copyright 1998 Annals of Botany Company Taxus mairei; ultrastructure; abscisic acid; gibberellin; seed dormancy; stratification; germination.     

A Mathematical Model to Utilize the Logistic Function in Germination and Seedling Growth

Several models have been proposed to describe germination rates,but most are limited in statistical analysis and biologicalmeaning of indices. Therefore, a mathematical model is proposedto utilize the logistic function. The function was defined asan overall response including time, temperature, and the interactionbetween time and temperature. Cumulative germination percentagesover time were used to develop the model. Germination tests were conducted on indiangrass (Sorghastrumnutans (L.) Nash) strain ‘IG-2C-F1’, at constanttemperatures of 9, 12, 15, 20, 25, and 30 °C. The functionfitted the observed data over six temperatures at r² = 0.99.Time to reach 10% of final germination (Gt₁₀) increased from2.5 d at 30 °C to 44.0 d at 9 °C, and Gt₅₀ (time toreach 50% of final germination) increased from 3.6 d at 30 °Cto 53.8 d at 9 °C. True germination rate (% d^–1) foreach temperature was maximum at Gt₅₀. A linear model of 1/Gt₅₀versus temperature was used to estimate the base temperatureof 8.3 °C for germination. An Arrhenius plot indicated achange occurred between 20 °C and 25 °C for temperatureresponse of germination. Published data on hypocotyl growthof Cucumis melo L. were recalculated using the model. Absolutegrowth rates showed a temperature response similar to the publishedweighted-mean elongation rates. Base temperature for hypocotylgrowth of C. melo was estimated as 8.8 °C. The proposedmodel proved to be useful in calculating and interpreting germinationand growth kinetics. Key words: Indiangrass, Sorghastrum nutans (L.) Nash, Germination rate, Threshold temperature, Arrhenius plot, Growth rate, Cucumis melo L     

Light, temperature and nitrogen as interacting factors affecting diel vertical migrations of dinoflagellates in culture

Diel vertical migrations of the marine dinoflagellates Gonyaulaxpolyedra Stein and Ceratium furca (Ehr.) Clap, et Lachm. werefollowed in a laboratory tube (2.02 m x 0.25 m) under a 12:12hlight:dark cycle. The effects of temperature stratification,two levels of surface irradiance and nitrogen depletion on patternsof vertical migrations were examined. At temperatures between22–26°C with small temperature gradients, both speciesmigrated at a rate of 0.7 –1.0 m h^–1. Steeper thermoclines(ca. 0.8°C 0.1 m^–1) with temperatures below ca. 20°Ccaused a marked decrease in swimming speed which resulted inaccumulations of cells in these thermocline regions. Under conditionsof nutrient sufficiency both algae migrated into the surfacelayers at irradiance values of over 1000 µE m^–2s^–1. Increasing nitrogen depletion caused the downwardmigration of both algae to commence progressively earlier inthe day and before the end of the light period. The earlierdownward migrations enabled a more complete descent throughthe thermocline. Nitrogen depleted cells of Gonyaulax continuedto undertake vertical migrations but avoided high irradiancesthus forming subsurface maxima at irradiance levels close to150 µE m^–2 s^–1. Ceratium cells which exhaustedboth inorganic nitrogen and phosphorus ceased to migrate accompaniedby a large change in cellular fluorescence.     

Theoretical Basis of Protocols for Seed Storage III. Optimum Moisture Contents for Pea Seeds Stored at Different Temperatures

In previous work, we demonstrated that there was an optimummoisture level for seed storage at a given temperature (Vertucciand Roos, 1990), and suggested, using thermodynamic considerations,that the optimum moisture content increased as the storage temperaturedecreased (Vertucci and Roos, 1993b). In this paper, we presentdata from a two year study of aging rates in pea (Pisum sativum)seeds supporting the hypothesis that the optimum moisture contentfor storage varies with temperature. Seed viability and vigourwere monitored during storage under dark or lighted conditionsat relative humidities between 1 and 90%, and temperatures between-5 and 65°C. The optimum moisture content varied from 0·015g H₂O g^-1 d.wt at 65°C to 0·101 g H₂O g^-1 d.wt at15°C under dark conditions and from 0·057 at 35°Cto 0·092 g H₂O g^-1 d.wt at -5°C under lighted conditions.Our results suggest that optimum moisture contents cannot beconsidered independently of temperature. This conclusion hasimportant implications for 'ultra-dry' and cryopreservationtechnologies.Copyright 1994, 1999 Academic Press Seed storage, seed aging, seed longevity, water content, temperature, glass, desiccation damage, ultradry, Pisum sativum L., pea, cryopreservation     

Effects of Irradiance and Temperature on Flowering of Chrysanthemum morifolium Ramat. in Continuous Light

The short-day plant Chrysanthemum morifolium cv. Polaris initiatedflower buds in all irradiances of continuous light from 7.5to 120 W m^–2. As the irradiance increased, the transitionto reproductive development began earlier and the number ofleaves initiated before the flower bud was reduced. The autumn-floweringcultivars Polaris and Bright Golden Anne, and the summer-floweringGolden Stardust were also grown in continuous light at differenttemperatures; all initiated flower buds at temperatures from10 to 28 °C but only the buds of Golden Stardust developedto anthesis and then only at 10 and 16°C. Flower initiationbegan earliest at 16–22 °C, and the number of leavesformed before the flower bud was increased at 28°C. GoldenStardust was exceptional in that the number of leaves formedwas also increased at 10 °C. Axillary meristems adjacentto the terminal meristem initiated flower buds rapidly at 10°C but not at 28 °C in all three cultivars. These resultsare discussed in relation to the autonomous induction of flowerinitiation and the effects of the natural environment on floweringof chrysanthemum. Chrysanthemum morifolium Ramat, flowering, irradiance, temperature     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.): VII. INFLUENCE OF AFTER-RIPENING AND AGEING ON GERMINATION RESPONSES TO TEMPERATURE AND WATER POTENTIAL

The effects of storage conditions on the germination of developingmuskmelon (Cucumis melo L.) seeds were tested to determine whetherafter-ripening is required to obtain maximum seed vigour. Seedswere harvested at 5 d intervals from 35 (immature) to 60 (fullymature) days after anthesis (DAA), washed, dried, and storedat water contents of 3·3 to 19% (dry weight basis) at6, 20, or 30°C for up to one year. Germination was testedin water and in polyethylene glycol 8000 solutions ( –0·2to –1·2 MPa osmotic potential) at 15, 20, 25 or30°C. Germination percentages and rates (inverse of meantimes to radicle emergence) were compared to those of newlyharvested, washed and dried seeds. For 40 and 60 DAA seeds,one year of storage at 20°C and water contents <6·5%significantly increased germination percentages and rates at20°C, but had little effect on germination at 25 and 30°C.Storage reduced the estimated base temperature (T_b) and meanbase water potential (_b) for germination of both 40 and 60 DAAseeds by approximately 5°C and 0·3 MPa, respectively.Immature 35 DAA seeds showed the greatest benefit from storageat 3 to 5% water content and 30°C, as germination percentagesand rates increased at all water potentials (). Storage underthese same conditions had little effect on the germination ofmature seeds in water, but increased germination percentagesand rates at reduced 's. Accelerated ageing for one month at30°C and water contents from 15 to 19° increased germinationrates and percentages of mature seeds at reduced 's, but longerdurations resulted in sharp declines in both parameters. Immatureseeds lost viability within one month under accelerated ageingconditions. An after-ripening period is required at all stagesof muskmelon seed development to expand the temperature andwater potential ranges allowing germination and to achieve maximumgerminability and vigour. Post-harvest dormancy is deepest atthe point of maximum seed dry weight accumulation and declinesthereafter, both in situ within the ripening fruit and duringdry storage. Key words: Muskmelon, Cucumis melo L., seed, development, dormancy, germination, vigour, after-ripening     

Androgenic Stimulating Factors in the Anther and Isolated Pollen Grain Culture of Datura innoxia Mill

Flower buds, anthers, and/or pollen grains collected at thetime of first haploid mitosis and 1–2 d before or afterthis division were submitted to different treatments beforeculturing anthers or isolated pollen grains. In the case ofanther culture, the percentages of androgenic anthem were notedat the end of 2, 3, 4, and 5 weeks of culture. Statistical studiesof the results thus obtained showed that some factors were highlyeffective in favouring androgenesis. The best results were obtained1–2 d after the first haploid mitosis with anther's centrifugedat 40 g for 5 min after cold treatment of the flower buds (48h at 3 °C); these treatments increased the percentage ofandrogenic pollen grains 12-fold. In case of isolated pollengrains, a system of culture particularly favourable for inductionand development of androgenic embryos was found. This systemincluded a cold treatment of the flower buds (48 h at 3 °C),the centrifugation of the isolated pollen grains (120 g for15 min), and culturing them for 20 d in the dark and then incontinuous light.