Reduction of sclerotial inoculum of Sclerotinia sclerotiorum with Coniothyrium minitans

Aspects of the biology of C. minitans and its potential for control of S. sclerotiorum were investigated.Temperatures below 7°C resulted in comparatively slow rates of germination and infection of sclerotia by C. minitans. The optimum temperature for germination, growth, infection of sclerotia, and destructive parasitism by C. minitans was 20°C. The optimum relative humidity for germination, growth and infection by C. minitans was above 95%.Autumn inoculations with suspensions of conidia, pycnidia and mycelium of C. minitans in the field resulted in negligible numbers of sclerotia remaining viable after 1 month. With culture-grown sclerotia 2 months were required for a similar reduction of sclerotial viability. In the absence of C. minitans mulching had no significant effect on sclerotial viability. In the presence of C. minitans mulching did, however, influence the viability and infection by C. minitans of culture-grown sclerotia. Populations of field sclerotia also differed from culture-grown sclerotia in that they harboured an internal population of microorganisms, which included C. minitans, and had a lower level of viability at the commencement of the treatments.A winter application of C. minitans did not result in significant infection of sclerotia nor in a reduction in viability of sclerotia. This failure is believed to have resulted from low temperatures and dry conditions.     

Some factors affecting survival of sclerotia of Macrophomina phaseolina in soil

At least 75% of the sclerotia of Macrophomina phaseolina survived for 1 yr in most natural soils kept at 26°C and at 50–55% of the soil moisture holding capacity (m.h.c.). Although survivability was reduced in a very acid soil (pH 4.5) collected under a pine stand, 33% of the sclerotia survived for 1 yr. Soil pH had very little or no effect on sclerotial survivability. Of three organic amendments tested (alfalfa hay, chitin, pine needles) only ground alfalfa hay at 0.8% (w/w) reduced survivability of sclerotia in soil by about 75% in a year. Alfalfa hay at 0.4% reduced survivability by 36%. Various N sources added at 200 μg Ng^?1 soil had no effect on survival. Of 13 fungicides tested, only benomyl and captan at 20 μg a.i. g^?1 soil appreciably reduced populations of sclerotia in soil.Soil temperature and moisture content were the two most important factors affecting survivability of sclerotia. At ?5 or 5°C the biggest drop in sclerotial survivability occurred when the soil was incubated moist (at 50% m.h.c. or more). At 26°C the biggest drop occurred in air-dried soil (2–3% m.h.c.) and survivability was decreased to some extent at 15 and 30% m.h.c. Survivability also dropped rapidly in moist soil (50–55% m.h.c.) exposed to four cycles each having 3-week freezing (?5°C) and 1 week thawing (26°C). Sclerotia in air-dried soil (2–3% m.h.c.) continuously kept at ?5°C maintained nearly complete survivability after 16 weeks. Sclerotia survived almost 80–90% in moist soil (50–55% m.h.c.) kept for 16 weeks at 26°C or in moist soil exposed to four cycles each having 3-week thawing (26°C) and 1-week freezing (?5°C).     

Carbon loss by sclerotia of Sclerotium rolfsii under the influence of soil pH,temperature and matric potential and its effect on sclerotial germination and virulence

Germinability and virulence of sclerotia of Sclerotium rolfsii were assessed after 50 days of exposure of ¹⁴C-labeled sclerotia to soil at 0, −5 and −15 kPa and pH 6.9, or to soil at 15, 25 or 30 °C, pH 5 or 8 and −1 kPa. Evolution of ¹⁴CO₂ accounted for the greatest share of endogenous carbon loss from sclerotia under all soil conditions, except in water-saturated soil (0 kPa), in which sclerotial exudates contributed the major share of carbon loss. Total evolution of ¹⁴CO₂ from sclerotia in soil at −15 kPa (42.4% of total ¹⁴C) and at −5 kPa (38%) was significantly higher than at 0 kPa (23.8%). Evolution of ¹⁴CO₂ in soil at 25 or 30 °C was more rapid than at 15 °C with regardless of pH. Loss of endogenous carbon by sclerotia was the greater after 50 days of exposure to soil at 0 kPa, or at 25 or 30 °C and pH 8, than at other soil conditions. Sclerotia exposed to water-saturated soil (0 kPa) showed a more rapid decline in nutrient independent germinability, viability and virulence, than to those exposed to −5 or −15 kPa. Sclerotia became dependent on nutrient for germination and lost viability and virulence within 30–40 days in soil at 25 or 30 °C, pH 8. However, more than 60% of sclerotia retained viability in soil at 15 °C regardless of pH, even after 50 days. Radish shoot growth was increased significantly by the sclerotia that had been exposed to soil at 0 kPa, or to soil at 25 or 30 °C and pH 8 for 50 days. In conclusion, carbon loss by sclerotia during incubation on soil at different pH levels, temperatures and water potentials was inversely correlated with sclerotial ability to infect radish seedlings. The relationship between carbon loss by sclerotia and radish shoot length was positive.     

Time-temperature relationships for the inactivation of sclerotia oF Sclerotium oryzae

The viability of sclerotia of Sclerotium oryzae was tested in both wet and dry soil under simulated elevated temperatures in either constant, cyclic or short temperature regimes. In wet soil viability was reduced to zero by a constant temperature of 45°C and above for 1 day or a minimum oven temperature of 55°C for 2 h. Viability of sclerotia were destroyed by either a 10-day constant temperature of 40°C or a 2-h temperature cycle for 3 days at 50°C or 12 days at 45°C. Below 40°C sclerotia were not affected. In dry soil at 60°C, a 3-day constant temperature or a 12 days 2-h temperature cycle eliminated sclerotial viability. Data on time and temperature relationships on loss in viability of sclerotia in soil could thus be used to predict efficacy of soil solarization.     

Polyethylene mulching of soil to reduce viability of sclerotia of Sclerotium oryzae

Mulching of Sclerotium oryzae infested soil (moist or dry) with polyethylene sheets during hot summer days of May and June increased the soil temperature at 5 cm from 36°C (unmulched) to 48°C (wet) and from 44 to 52°C (dry) and at 20cm from 32 to 38°C (wet) and from 35 to 39°C (dry). In artificially-infested soil, the sclerotia were not eradicated but 95–100% loss in viability was observed at 5 cm by a mulch treatment for 1 week and at 20 cm by mulching for 8 weeks. Mulching effects were not influenced by moisture content of soil or by amendments with lucerne or wheat straw. Mulching of naturally-infested soil at a second site did not eradicate S. oryzae but reduced sclerotial viability by 93%.     

Interactions between Sclerotium rolfsii and Trichoderma spp: Relationship between antagonism and disease control

Ten isolates of Trichoderma spp were examined for their ability to antagonize growth and to parasitize mycelium of Sclerotium rolfsii (Sr-1) on agar media, to inhibit germination of sclerotia of S. rolfsii on natural soil plates and to sporulate on the sclerotia, and to protect bean seedlings against the pathogen in the greenhouse. A high negative correlation (r = ?0.844) was observed between plant stand in the greenhouse and sclerotial germination on soil plates but not with antagonism on agar plates. Three isolates of T. harzianum (Th-7, Th-20, WT-6) and one of T. hamatum (TRI-4) were especially effective in reducing sclerotial germination and controlling disease in the greenhouse. Three isolates of Trichoderma spp (WT-6, TMP, and TRI-4), effective in reducing sclerotial germination of isolate Sr-1, also prevented sclerotial germination in four out of five additional S. rolfsii isolates studied.     

Population and distribution of sclerotia of Rhizoctonia solani kühn in sugar beet field soil

The population and distribution of sclerotia of Rhizoctonia solani Kühn in two sugar beet field soils was determined at harvest by a sieving-flotation method. In rhizosphere soil (RS) and non-rhizosphere soil (NRS) from the most heavily infected roots of sugar beets, 1.43–2.5 and 0.83–1.0 sclerotia g^?1 dry soil were detected, respectively. In the soil around healthy sugar beet, these values were 0.04–0.12 and 0.03–0.04 sclerotia g^?1 dry soil. More sclerotia were always obtained from RS than from NRS. More than 80% of the sclerotia were in the upper 10 cm of soil and within 10 cm of diseased roots. Therefore, there is a non-uniform distribution of sclerotia of R. solani in soil.The sclerotial population in soil increased significantly with disease severity and a good correlation was obtained between the number of sclerotia and the disease severity on infected plants. Most of the sclerotia collected from the field soil ranged in size from 0.5 to 2.0 mm diameter.Viability of sclerotia increased as severity of crown rot increased and as the size of the sclerotia increased. Conversely, there was a progressive decrease in sclerotial germination with increasing depth in soil and increasing distance from the infected root.     

Reduction in viability of sclerotia of Macrophomina phaseolina with polyethylene mulching of soil

Mulching of Macrophomina phaseolina-inksted soil (moist or dry) with transparent polyethylene sheets during the hot days of May increased temperature of wet soil at 5 cm from 37°C (unmulched) to 52°C (mulched) and of dry soil from 52°C (unmulched) to 65°C (mulched). At 20 cm mulching increased temperature from 30°C to 41°C (wet) and from 38°C to 42°C (dry). In artificially-infested soil. the sclerotia of M. phaseolina were eradicated at 5 cm by a mulch treatment for 1 week and at 20 cm depth 50% sclerotia lost viability in wet soil but were not affected in dry soil. In a naturally infested soil (5–7 sclerotia g^?1), which gave 20% infection on Vigna, the sclerotia were reduced to such an extent that after 1 week mulching no disease was observed on Vigna.     

Microbial populations involved in the suppression of Rhizoctonia solani AG1-1B by lignin incorporation in soil

Rhizoctonia solani causes worldwide losses in numerous crops. Sclerotia of R. solani remain viable for several years in soil and are an important source of primary infection. In this study the effect of soil incorporation of Kraft pine lignin, a side product of the paper industry, on viability of R. solani AG1-1B sclerotia was investigated. The efficacy of lignin was assessed in a sandy loam (Oppuurs) and a silt loam soil (Leest) collected from commercial fields in Belgium. Evaluating sclerotial viability after 4 weeks incubation in the two soils amended with 1% (w/w) Kraft pine lignin demonstrated a soil-dependent effect. In Leest soil the addition of lignin resulted in a significantly reduced sclerotial viability, together with an increased mycoparasitism by Trichoderma spp.; in Oppuurs soil, on the other hand, only a slight and insignificant reduction in sclerotial viability was observed. Based on phospholipid fatty acid analysis, different changes in microbial community structure upon lignin amendment were detected in the two soils. Both amended soils showed a significant increase in Gram negative bacteria. In Leest soil this increase was accompanied with a significantly higher increase in fungi and actinomycetes compared with Oppuurs soil. In addition, Kraft pine lignin resulted in both soils in a small but significant increase in manganese peroxidase activity and this increase tended to be higher in Leest soil. Manganese peroxidase produced by lignin-degrading basidiomycetes has previously been shown to degrade melanin, which protects the sclerotia against biotic and abiotic stress. We hypothesize that lignin-degrading fungi increased the susceptibility of the sclerotia to sclerotial antagonists such as Trichoderma, Gram negative bacteria and actinomycetes. Clearly, the effect observed here did not rely on the stimulation of one microbial group, but is the result of an interaction of different groups.     

Diversity of potential microbial parasites colonizing sclerotia of Macrophomina phaseolina in soil

The colonization of Macrophomina phaseolina sclerotia by microbial parasites was evaluated in unsterilized field soil at different levels of soil moisture (0,-5, and-10 kPa) and temperature (20, 30, and 40°C). The maximum colonization of sclerotia was recorded in soil held at-5 or-10 kPa at 30–40°C. Trichoderma harzianum isolate 25–92 and Pseudomonas fluorescens isolate 4–92 were recorded as potential sclerotial parasites, and they significantly (P=0.05) reduced the germination of sclerotia by 60–63%. Cells of P. fluorescens and buffer-washed conidia of T. harzianum were completely agglutinated at 28°C with crude agglutinin of M. phaseolina. The ability of different antagonists to parasitize the sclerotia were correlated with the agglutination ability of the antagonists.     

The effect of dry conditions on subsequent leakage and rotting of fungal sclerotia

Dried sclerotia of Sclerotium delphinii rotted in moist soil whereas those of Sclerotium cepivorum. Botrytis cinerea and Botrytis tulipae did not. A number of fungi invaded dried sclerotia of S. delphinii in soil, the principal coloniser found in the first sampling being Trichodermu hamatum. Leakage of ¹⁴C compounds from dried labelled sclerotia placed in water was rapid and was little affected by variation in leakage temperature from 1 to 25°C or by prolonging the drying period beyond a day. Leakage from dried sclerotia which were allowed to imbibe water through a small part of their surface was much reduced. Sclerotia which were redried after leakage leaked again when returned to water but with all four fungi the first of three leakage cycles gave the highest ¹⁴C levels. Loss in dry weight in the first leakage cycle was greater with S. delphinii than with the other three fungi and this may be related to the poor survival of dried sclerotia of S. delphinii in moist soil. Substances lost during leakage appear to originate from within sclerotial hyphae rather than from the hyphal free space.     

Multivariate effects of plant canopy, soil physico-chemistry and microbiology on Sclerotinia stem rot of soybean in relation to crop rotation and urban compost amendment

The effects of canopy, soil physico-chemical and microbiological variables on Sclerotinia stem rot (SSR) on soybean were assessed in two soils (clay loam and sandy loam) using multiple regression and canonical redundancy analysis (RDA) and their partial form to control for the rotation (2 or 3-y-corn/soybean monoculture) and fertilization (mineral/urban compost) or spatial variables effects. The models revealed the minimal sets of variables that best explain the variance of the survival of Sclerotinia sclerotiorum’s sclerotia, carpogenic germination, disease severity and their associations. In clay loam, the 3-y-corn rotation reduced disease severity mainly through the reduction of weed biomass that favoured carpogenic germination. Urban compost has a conducive effect explained by a better soil surface drainage. Additionally, total N was found suppressive to sclerotial survival. In sandy loam, the carpogenic germination was negatively correlated with high C mineralization quotient and aggregate stability but correlated positively with Ca. Sclerotial survival was negatively correlated with pH and Ca, and positively correlated with biological fertility index. Aggregate stability, Ca and pH were associated with the urban compost. The regression and RDA analyses allowed to identify key variables that drived SSR development and explain their relationship with the cultural practices, soil health, as well as the spatial variation of disease variables.     

Survival of Coniothyrium minitans associated with sclerotia of Sclerotinia sclerotiorum in soil

The development and survival of the mycoparasite Coniothyrium minitans associated with sclerotia of the plant pathogen Sclerotinia sclerotiorum was studied in pasteurised and non-sterile (untreated) soil. Using scanning electron microscopy, developing pycnidia were first seen within the sclerotial medulla at 7 days post-inoculation with the mycoparasite in pasteurised soil. However, by 14 days post-inoculation, pycnidia had developed fully in both pasteurised and non-pasteurised treatments, and conidial droplets were exuded onto the outer surface of the infected sclerotia. Thirty days post-inoculation, irrespective of soil treatment, the majority of the sclerotial medulla had been converted to pycnidia, with the sclerotial rind remaining largely intact. The pycnidia and dried intact droplets were still observed 6 months post-inoculation with C. minitans, although the conidia on the outer surface of the dried droplets had largely collapsed by this stage. Germinability studies at 10 months post-inoculation showed that approximately 13% of the conidia in dried droplets were still viable. This work shows the potential for infected sclerotia of S. sclerotiorum to provide a unique reservoir for the survival of C. minitans.     

Time and burial depth influencing the viability and bacterial colonization of sclerotia of Sclerotinia sclerotiorum

Sclerotia are the primary over wintering inoculum of Sclerotinia sclerotiorum (Lib.) de Bary. The effects of tillage on the primary inoculum are not well understood. The purpose of this research was to study sclerotial viability over time and between burial depths in soil, to identify bacteria colonizing and degrading the sclerotia, and determine whether these bacteria may be utilized as biological control agents. Correlation analysis indicated that a significant negative relationship existed between sclerotial viability and elapsed temporal factors (R²=−0.68, P<0.0001), and depth of burial (R²=−0.58, P<0.0001). After twelve months, sclerotia on the soil surface had the highest viability (57.5%), followed by those at the 5 cm depth (12.5%), and only 2.5% of those placed at the 10 cm depth remained viable. A significant negative relationship between sclerotial viability and bacterial populations also existed (R²=−0.60, P<0.0001). Two hundred and sixty-eight bacteria were isolated from sclerotia, 29 of which showed strong in vitro antagonism to the mycelial growth of S. sclerotiorum. Biodiversity of the inhibitory bacterial isolates was minimal on sclerotia from the soil surface and within all depths sampled at three months (i.e. in January). All burial depths within the April and July sampling dates produced bacterial diversities that were distinct from each other.     

Loss of nutrient-independence for germination by fungal propagules incubated on soils or on a model system imposing diffusive stress

Nutrient independent conidia of Cochliobolus victoriae and sclerotia of Sclerotium cepivorum, Macrophomina phaseolina, and Verticillium dahliae were incubated aseptically on sand through which a dilute salts solution percolated at a flow rate sufficient to inhibit germination. Propagules were then transferred to a static salts solution to assess germination. Conidia of C. victoriae and sclerotia of S cepivorum became nutrient-dependent (6% germination in salts solution) after 9 and 15 days on sand, respectively. Thirty-five days of diffusive stress were required to attenuate the nutrient-independence of M. phaseolina sclerotia. Sclerotia of V. dahliae lost little of their nutrient-independence even after 45 days of diffusive stress. Viability of C. victoriae and S. cepivorum was reduced after 45 days of diffusive stress, but viability of V. dahliae and M. phaseolina was not. Conidia of C. victoriae gradually became nutrient-dependent when incubated for several weeks on each of five soils. A loam and two sandy loam soils were more effective in decreasing the nutrient-independence of conidia than were two clay loams. Sclerotia of M. phaseolina also lost nutrient-independence when incubated on four of the five soils.Interruption of artificially imposed diffusive stress resulted in increased [¹⁴C]exudation from conidia of C. victoriae and sclerotia of M. phaseolina. Germinability on salts solution of C. victoriae conidia previously made nutrient-dependent was significantly increased, when the conidia were kept at 4°C for 3.5 days before germination assay. Conidia of C. victoriae made nutrient-dependent and then incubated on soils labeled with [¹⁴C]glucose, absorbed twice as much ¹⁴C from a loam and two sandy loam soils as from two clay loam soils. Following incubation on four of the five ¹⁴C-labeled soils, the germinability of the conidia in the absence of nutrients was significantly increased over that of conidia not incubated on these soils.The results suggest that a continued minimal stress may be needed to maintain the nutrient-dependence of some fungal propagules in soil. Interruption of nutrient stress appears to allow nutrient-dependent propagules an opportunity to recoup nutrients from the soil solution or to reorganize endogenous energy reserves whereby the potential for germination is increased.     

Soil temperature,mycorrhizal infection and nodulation of Medicago truncatula and Trifolium subterraneum

Establishment of vesicular-arbuscular mycorrhizal fungi in plant roots involves a pre-infection phase of propagule germination, hyphal growth and appressorium formation, followed by growth of the fungus within the root. The effect of soil temperature on the pre-infection stage was examined by counting the numbers of fungal “entry-points” on the main roots of Medicago truncatula and Trifolium subterraneum, grown at soil temperatures of 12°, 16°, 20° and 25°C for periods up to 12 days. Increased root temperature was positively associated with increased numbers of “entry-points”. This effect was more marked between 12° and 16°C than at higher temperatures, as shown by comparing plants at the same stage of development (emergence of spade leaf) and by calculating the results as entry points per cm root.The first root nodules appeared sooner at higher temperatures (20° and 25°), but subsequent development of nodules (measured as nodule number and aggregate volume of nodules per plant, up to 21 days) was best at 16°C for both host Rhizobium combinations in non-sterile and autoclaved soil. There was no evidence that competition between mycorrhizal fungi and Rhizobium for infection sites occurred.A method of obtaining numbers of infective propagules of vesicular-arbuscular mycorrhizal fungi in soil is described.     

Evaluation of different Coniothyrium minitans inoculum sources and application rates on apothecial production and infection of Sclerotinia sclerotiorum sclerotia

The effects of three Coniothyrium minitans isolates (Conio, IVT1 and Contans^®), applied to soil as conidial suspensions or as maizemeal-perlite (MP) inocula (Conio), on apothecial production and infection of Sclerotinia sclerotiorum sclerotia were assessed in two soil pot bioassays and two novel box bioassays in the glasshouse at different times of the year. C. minitans isolate Conio applied as either MP or ground MP at full rate (10⁶-10⁷ cfu cm⁻³ soil) consistently decreased the carpogenic germination, recovery and viability of sclerotia and increased C. minitans infection of the sclerotia of S. sclerotiorum by in comparison with either MP or conidial suspension treatments applied at lower rates (10³-10⁴ cfu cm⁻³ soil). Additionally, when applied at the same rate, MP inoculum of C. minitans was consistently more effective at reducing carpogenic germination than a conidial suspension. The effect of MP and ground MP at full rate on carpogenic germination was expressed relatively early as those sclerotia recovered before apothecia appeared on the soil surface already had reduced numbers of apothecial initials. In general, there were few differences between the isolates of C. minitans applied as conidial suspensions. Box bioassays carried out at different times of the year indicated that temperature and soil moisture influenced both apothecial production and mycoparasitism. Inoculum concentration of C. minitans and time of application appear to be important factors in reducting apothecial production by S. sclerotiorum.     

Comparaison des aptitudes parasitaires de clones de Trichoderma vis-a-vis de quelques champignons a sclerotes

Nineteen monoconidial isolates (referred to as clones) of Trichoderma from different species aggregates, one isolate of Gliocladium virens, and one isolate of an Acrostalagmus sp. (that was naturally associated with sclerotia of Sclerotinia spp and Macrophomina phaseolina) were tested. They were incubated in controlled conditions, in sterile soil, with sclerotia of Corticium rolfsii, Sclerotinia minor, or S. sclerotiorum. At the end of appropriate periods of incubation (respectively 26, 20 and 8 days), the sclerotia were retrieved from soil and checked for invasion by the antagonist. Important differences between the parasitic ability of Trichoderma clones were noted. Clones from at least three different species (T. aureoviride, T. hamatum, T. harzianum) exhibited a high antagonistic activity. Activity of the G. virens isolate was at the same level as the best clones of Trichoderma, whereas no parasitic tendencies were found in the isolate of Acrostalagmus sp., thus confirming previous results.A rather good correlation was found between the capacity of the clones for attacking C. rolfsii sclerotia and their ability to parasitize both Sclerotinia.In conclusion, it is proposed that a screening with only one of the sclerotial species would give clones efficient against all three, and possibly against related sclerotial types.     

Survival of sclerotia of Macrophomina phaseolina and Sclerotium cepivorum after drying and wetting treatments

Exposure of sclerotia of Macrophomina phaseolina to 0 and 33% relative humidity (r.h.) for 12 weeks and of Sclerotium cepivorum to 0, 33 and 55% r.h. for 20 weeks did not reduce their germinability on agar. Exposure to 78% r.h. caused high loss of germinability in M. phaseolina and complete loss in S. cepivorum. After 7-day exposures respective moisture contents of sclerotia of M. phaseolina and S. cepivorum were 1 and 2% at 0% r.h.; and 10 and 14% at 78% r.h. M. phaseolina sclerotia held at 0% and 33% r.h. in desiccators for several times up to 12 days did not decrease in subsequent survivability in moist soil, unlike sclerotia held at 78% r.h. for 4 days.More sclerotia of M. phaseolina were colonized by fungi and Streptomyces spp. on alkaline soil than on acid soil. On alkaline soil twice as many sclerotia were colonized after exposure to 0% r.h. as after exposure to 33, 55 and 78% r.h. Colonization of S. cepivorum sclerotia was as high on acid as on alkaline soil and 3 times as high on sclerotia treated at 0% r.h. as on those treated at higher r.h. Attempts to ascertain the effects of colonization on sclerotial viability were unsuccessful. Incubation of sclerotia of M. phaseolina in moist Rumsford sandy loam (50% m.h.c.) for 20 weeks reduced survivability by 43%. At room temperature, alternate drying and wetting of soil containing sclerotia did not appreciably affect survivability of either pathogen. Survivability of S. cepivorum sclerotia was highest when the sclerotia were incubated in air-dried soil (2–3% m.h.c.) for 20 weeks.Incidence of white rot on onion seedlings transplanted to S. cepivorum-infested soil was higher in soil that had been air-dried for 20 weeks than in soil that had been alternately wetted and dried. Sclerotia that were exposed to 0% r.h. for 7 days before soil incubation produced little white rot.     

Effects of light and temperature on seed germination of Poa pratensis from high latitudes

One‐to‐three‐year‐old seed of Poa pratensis "Lavang”;, “Leikra”; and “Ryss”; was germinated under various temperature and light regimes on a thermogradient plate and on paper or in soil in phytotron compartments. While the optimal constant temperature for germination of Lavang and Leikra in 12 h light/12 h dark cycles was around 16°C, Ryss germinated nearly 100% over the 10–28°C temperature range. Compared with constant temperatures, daily fluctuations, even at the small amplitude 18/15°C, stimulated germination of Lavang and Leikra. Dark germination was inferior to germination in light/ dark cycles at constant temperatures of 15°C and higher, but light showed no advantage at alternating temperature. Continuous light inhibited germination regardless of light source (fluorescent, incandescent or natural). The stimulating effect of daily light/dark cycles increased, but the inhibiting effect of continuous light decreased with increasing seed age. It is concluded that seed of Poa pratensis ought to be covered by a 0.5–1.0 cm soil layer when sown in a northern environment involving continuous light and low soil temperatures.