Alfalfa seed germination tests and stand establishment: The role of hard (water impermeable) seed

John W. Hall1, Darryl G. Stout2, and Barbara M. Brooke2

1Research

Centre, Agriculture and Agri-Food Canada, Pacific Agri-Food Research Centre, Summerland, British Columbia, Canada V0H 1Z0; and 2Range Research Unit, Agriculture and Agri-Food Canada, 3015 Ord Rd., Kamloops, British Columbia, Canada, V2B 8A9. Agriculture Canada contribution no. 1004. Received 14 February 1997, accepted 3 November 1997.
Hall, J. W., Stout, D. G. and Brooke, B. M. 1998. Alfalfa seed germination tests and stand establishment: The role of hard (water impermeable) seed. Can. J. Plant Sci. 78: 295300. Freeze thaw scarification has been observed to increase the germination rate of alfalfa (Medicago sativa L.) seed containing a large proportion of hard (water impermeable) seed in a 7-d laboratory germination test; however, a comparable increase in plant density is not always seen in the field. To investigate this discrepancy, a field experiment with untreated and scarified seed was carried out using cultivars with high (Apica, 35%; Barrier, 32%) and low (Apollo II, 1%; WL316, 0.3%) percentages of hard seed. Plants were counted at the three-trifoliate-leaf and 10% bloom stages in 1992, the planting year, and at 10% bloom in 1993. In the field, effects of scarification were seen only at the 10% bloom stage in 1992, increasing the plant densities of high hard seed cultivars by 17% while decreasing those of low hard seed cultivars by 10%. Two laboratory experiments were also done to determine the effect of temperature, lighting (light, shade, dark) and media (on blotter, in soil) on the germination of hard seed. Germination was highest at alternating 35/5C and higher at 28/14C than at constant 20C. At 20/10C most germinated seed died. Differences among germination media and lighting treatments were small. Scarification may possibly have some negative effect on the quick-germinating fraction of the seed in the field but the net effect for cultivars with a high hard seed content appears positive. Some seed identified as hard in a 7-d test at constant 20C likely germinate and contribute to an alfalfa stand in the field when soil temperatures are higher. Thus the discrepancy previously observed between the laboratory and field results may possibly be accounted for in part by a negative effect of scarification and also by differences in temperature. Key words: Medicago, germination, seed coat, temperature Hall, J. W., Stout, D. G. et Brooke, B. M. 1998. Essais de germination des graines de luzerne et installation : rle des graines dures (impermables leau). can. j. Plant Sci. 78: 295300. Un essai de germination en laboratoire de 7 jours a permis de constater que la scarification par alternance gel-dgel accrot le pouvoir germinatif des graines de luzerne (Medicago sativa L.) contenant une forte proportion de graines dures (impermables leau). Cependant cet accroissement ne semble pas toujours se rpercuter sur la densit de peuplement au champ. Pour lucider cette apparente divergence, une exprience au champ utilisant des graines non traites et des graines scarifies tait ralise sur des cultivars taux lev (Apica : 35 %, Barrier : 32 %) et faible (Appollo II : 1 %, WL316 : 0,3 %) de graines dures. Le dnombrement des plantes tait fait au stade de 3 feuilles trifolies ainsi quau stade de floraison 10 % en 1992 lanne de semis et de floraison 20 % en 1993. Au champ, les effets de la scarification ne se voyaient quau stade de floraison 10 % en 1992. La densit de peuplement de varits forte proportion de graines dures marquait une augmentation de 17 % tandis que celle des varits faible proportion de graines dures diminuait de 10 %. Deux expriences complmentaires en laboratoire ont servi dterminer leffet de la temprature, de lclairement (clair, ombrag, sombre) et du substrat (buvard ou sol) sur la germination des graines dures. Le pourcentage de germination le plus haut tait obtenu sous une alternance de 35 et 5 C. Il tait galement plus lev en alternance 28 C/14 C qu une temprature constante de 20 C. Sous alternance 20/10 C, la plupart des graines germes mouraient. Les diffrences dues au substrat de germination et au rgime dclairement taient de faibles amplitudes. La scarification pourrait avoir des effets ngatifs au champ sur la fraction germination rapide des graines, mais dans lensemble leffet net pour les cultures forte proportion des graines dures apparat positif. Une certaine quantit de graines identifies comme dures au test de germination de 7 jours temprature constante de 20 C ont des chances de germer au champ et denrichir le peuplement de luzerne lorsque le sol se rchauffe. Ainsi les carts observs entre les comportements des graines en laboratoire et au champ sexpliqueraient en partie par leffet ngatif de la scarification, de mme que par les diffrences de temprature. Mots cls: Medicago, germination, tgument sminal, temprature

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Viable alfalfa seed may be divided by a standard 7-d laboratory germination test into non-hard, or quick-germinating seed, which produces normal seedlings, and hard seed, which does not take up water during the 7 d. For legumes, percent PLS is calculated by summing percent quick-germinating seed and percent hard seed (Cooper 1977). There is a debate as to whether stand establishment is related to percent PLS or to percent quick-germinating seed. Bass et al. (1988) stated
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that hard seeds have little value when planted because these late-germinating seeds seldom contribute to improving the stand. Others have concluded that hard seeds are nearly

Abbreviations: D, plant density; dD differential change in plant density; dR, differential change in sowing rate; PLS, pure live seed; R, sowing rate

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as important in stand establishment as quick-germinating seeds (Leggatt 1927, 1930; Weihing 1940). Hall et al. (1993) reported that freeze-thaw scarification of Apica alfalfa increased germination in the 7-d laboratory test by 86% (from 50 to 93%) while plant density in the field was increased by only 40%. Several explanations for this discrepancy might be suggested. Possibly, the scarification treatment may have injured some seeds; the injury not being manifested until a later growth stage than that observed in a 7-d germination test. Another possibility is that more of the hard seed may have germinated in the field than in the laboratory, increasing the plant density in the control plots and thus decreasing the advantage conferred by scarification. There is some evidence that hard seeds can be genetically different from quick-germinating seeds. For example, average heights of plants arising from hard seed and from the original unsorted seed were similar for two alfalfa cultivars, but shorter from hard seed than from original seed for a third cultivar (Smith 1961). For all three cultivars winter injury was similar for plants from hard or original seed. Thus it is also possible that freeze-thaw scarification may affect each type of seed differently. On the other hand, this discrepancy may show that the standard germination test, which is conducted at constant 20C for alfalfa (Canada Department of Agriculture 1979), is not satisfactory for predicting germination under the variable environmental conditions experienced in the field; that is, seeds that do not imbibe water at 20C may do so at variable temperatures. This study was conducted to examine further the discrepancy between the 7-d germination test and field results reported by Hall et al. (1993). A field experiment that included cultivars with high and low percentages of hard seed was conducted to examine the effects of the scarification treatment. Laboratory tests were done with alternating temperatures that resembled field germination conditions to determine the effect on the germination of hard seed. Germination tests were also done on the seed used in the field experiment to find out if the germination rates of scarified seed change with time in storage. MATERIALS AND METHODS Field Experiment CULTIVAR SELECTION AND SEED TREATMENT. Four alfalfa cultivars were chosen for this experiment. Seed of two cultivars with high hard seed content, Apica and Barrier, was produced in Alberta in 1990, and was commercially cleaned but not scarified. It was stored during the winter at Kamloops in cloth bags at room temperature. Seed of two cultivars with low hard seed content, Apollo II and WL316, had been produced before 1990; the location and year being unknown. The Apollo II had a barely perceptible clay coating while the WL316 had a 33% lime coating. Subsamples of the four cultivars were freeze-scarified (Stout 1990) in April 1992. This treatment involved five freeze-thaw cycles, of 2 h at 80C in an Ultra Low Bio Freezer (Forma Scientific, Marietta, OH), then 2 h minimum at 20C.

STANDARD 7-DAY GERMINATION TEST TO ADJUST SEEDING RATES. Before planting, standard 7-d germination tests at 20C (Canada Department of Agriculture 1979) were conducted on the original untreated seed for seeding rate calculations. Tests for Apica and Barrier started 26 November 1990 and those for WL316 and Apollo II started 2 April 1992. The 100-seed weight was determined for each cultivar prior to this test, so that unbiased samples could be weighed, rather than counted out, for testing. The coating on the WL316 seed was removed after weighing, by enclosing the sample in open-weave fabric and rinsing and gently rubbing under cold running water. Seeds were incubated on standard blue blotters in petri plates (two plates of approximately 100 seeds each per cultivar) for 7 d in a 20C incubator. After the field experiment had been planted, 7-d germination tests of both untreated and freeze-thaw scarified seed of each cultivar were carried out in the laboratory on four dates: 5 May, 16 June, 24 November and 11 December 1992. Three replicates of approximately 100 seeds each were tested for each of the eight cultivar-treatment combinations on each date. FIELD PLANTING AND OBSERVATIONS. The irrigated site at the Kamloops, British Columbia, Range Research Ranch (lat. 5042N, long. 12024W, elevation 349 m) was on an Orthic Humic Gleysol, fine silty over coarse loamy, mixed, alkaline, moderately warm subarid soil located in the Thompson River Valley bottom (van Ryswyk et al. 1993). The surface layer averages 7.5 pH (H2O), 4.3% organic matter (dichromate oxidation), and 0.54 dS m1 electrical conductivity (paste). Average annual precipitation is 257 mm, and 76 mm (water equivalent) of this annual precipitation occurs as snow. The average annual water deficit is 387 mm yr1. These weather data were obtained at Kamloops Airport (Environment Canada 1982) which is about 1 km south of the research site and at a similar elevation. The land for the experiment had been fallow for at least 2 yr. Three successive identical seeding trials were done on 30 April, 10 June and 30 July 1992. Seeding was carried out at a depth of 6 mm and a rate of 5 kg ha1 PLS using a one-row double-disk-opener plot seeder. Apica had the highest viability in the laboratory germination test so seeding rates for the other cultivars were adjusted to provide the same number of pure live seeds per unit area as 5 kg ha1 of Apica did. Plots consisted of two 6.7-m rows spaced 30 cm apart. Dry rhizobium inoculum (Nitragin Ltd., Milwaukee, WI), was applied to seeds immediately before seeding. The two seed treatments (control and freeze-thaw scarified) and four cultivars were seeded in a randomized complete block design with four blocks. One block was lost from the 30 April seeding. Plots were sprinkler irrigated with 20 mm of water during stand establishment. Additional irrigations of 50 mm were applied as needed throughout the season. On each observation date, plant counts were made for each plot in three 60-cm subplots within the same row and 180 cm apart. Plant counts were made by digging up plants from the three 60-cm subplots at the three-trifoliate-leaf stage (2 June, 7 July, and 3 September for trials one to three, respectively) and at the 10% bloom stage (78 July, 21

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August, and 67 October for trials one to three, respectively). A final count was made for all three trials at the 10% bloom stage during the period of 915 June 1993. Plant densities were expressed as a percentage by dividing the plant counts by the estimated number of pure live seed in 180 cm of row. Modified Germination Tests LABORATORY EXPERIMENT ONE. Clean Apica and Barrier seed from the 1989 crop was obtained from Bantry Seed Cleaners, Tilley Alberta. Three plates of approximately 100 seeds each were used for a 7-d germination test (Canada Department of Agriculture 1979) at 20C to determine percent hard seed. The weights for these lots were 2.11 and 2.05 mg seed1, and the hard seed contents were 37% and 23% for Apica and Barrier, respectively. Four months before the start of the experiment the hard seed was isolated by incubating all the seed on moist paper towel for 7 d and discarding all the non-hard seed. The remaining hard seed was dried by blotting with paper towel and then stored at room temperature in a glass bottle with a mesh top. Beginning on 28 January 1991, the hard seed was incubated on moist blue blotters for 6 wk, in the dark, under five temperature regimes: 12 h at 10C and 12 h at 20C, constant 5C, constant 20C, 12 h at 5C and 12 h at 35C, and constant 35C. Three petri plates of each cultivar with approximately 100 seeds per plate were exposed to each temperature treatment. At 1-wk intervals germinated seeds were counted and removed. Seeds with radicle length of at least 2 mm were considered germinated. At the end of 6 wk, dead and ungerminated fresh seeds were counted. LABORATORY EXPERIMENT TWO. Uncleaned Apica alfalfa seed from the 1990 seed crop was obtained from the Alberta Wheat Pool. Two plates with approximately 100 seeds each were used for a 7-d germination test at the start of the experiment. The material had 21% hard seed on 11 December 1992, and weighed 2.26 mg seed1. The remaining seed was incubated on moist paper towel for 7 d and all non-hard seed was discarded. The hard seed was then incubated under various combinations of three temperature and three lighting regimes and two media. The temperature regimes were constant 20C, 16 h at 14C followed by 8 h at 28C and 16 h at 5C followed by 8 h at 35C. The lighting regimes were darkness, 8 h light and 8 h light with a paper bag shade draped over the tray. Light was provided by two fluorescent and two incandescent lamps during the high temperature phase of the temperature regime. The two germination media were moist blue blotters on petri plates and moist sieved loam soil in pots. In a few cases Whatman #1 filter paper was used instead of blue blotter. Seedlings emerged from the soil, and those on the blotters having radicles at least 2 mm long were counted for each treatment at 7-d intervals for 8 wk. Statistical Analysis FIELD EXPERIMENT. The data from the 7-d germination tests carried out after the field experiment had been planted, had a 4 2 4 factorial structure with four cultivars, two levels of scarification (with and without) and four dates of testing.

Table 1. Percentage of hard (water impermeable) seed for the four alfalfa cultivars used in the 1992 field experiment % hard seed Cultivar Apica Barrier Apollo II WL316
zStandard

Untreated 35.21.4z 32.01.3 1.00.3 0.30.2 error.

Scarified 1.10.3 1.10.3 0.20.1 0.10.1

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The statistical analysis was carried out by fitting log-linear models estimated by maximum likelihood (Bishop et al. 1975). The field experiment was a 3 4 2 factorial with three trials, four cultivars and two scarification treatments. The effects of these factors and their interactions were determined by analysis of variance. The effects of cultivars were partitioned into the difference between cultivar types (high or low proportion of hard seed) and the differences between cultivars within types. All computations were performed using the SAS statistical package (SAS Institute, Inc. 1990). LABORATORY EXPERIMENTS. The germination data were analysed using log linear models estimated by maximum likelihood. Laboratory experiment 1 was a 2 5 factorial with two cultivars and five temperature regimes. Laboratory experiment 2 was not a complete factorial but had three factors: temperature, lighting and media. Analyses were carried out on the 4 and 8 wk data. These times were chosen because in the field experiment they were the time intervals from planting to the three-trifoliate-leaf and 10% bloom stages, respectively. All computations were performed using the SAS statistical package (SAS Institute, Inc. 1990). RESULTS Standard 7-d Germination Test For Field Experiment There were differences among cultivars in the proportion of viable seed that was hard. Viable Apica and Barrier seed contained 35 and 32% hard seed, respectively, while viable Apollo II and WL316 seed contained 1% or less of hard seed (Table 1). After freeze-thaw scarification all four cultivars had hard seed contents of 1% or less. There was no change in germination rate of either untreated or scarified seed between 5 May and 11 December (data not shown). Plant Density in the Field Plant density varied between cultivars within type (high or low hard seed) (Table 2). In the unscarified seed, high hard seed cultivars had higher plant densities at the three-trifoliateleaf stage than low hard seed cultivars. At the 10% bloom stage in 1992, the plant density of WL316 was similar to that of the high hard seed cultivars while the plant density of Apollo II was higher. In the spring of 1993 this pattern was reversed and Apollo II was similar to the high hard seed cultivars while WL316 had a lower plant density. Freeze-thaw scarification had no detectable effect either at the three-trifoliate-leaf stage or in the spring of 1993 (Table 2). At the 10% bloom stage in 1992, scarification

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Table 2. Effect of hard seed content and scarification on plant density expressed as a percentage of viable seed sown in 1992 Hard seed content before scarification (HSC) High Low SE 10% bloom stage, 1992 High Low Apica Barrier Apollo II WL316 SE High Low SE Source Trial Block (trial) HSC Cv (HSC) S HSC S Cv S (HSC) HSC Trial Cv (HSC) Trial S Trial S HSC Trial Error df 2 8 1 2 1 1 2 2 4 2 2 56 Apica Barrier Apollo II WL316 10% bloom stage, 1993 43.0 46.1 47.7 39.0 3.10 Analysis of variance mean squares 3-trifoliate leaf stage, 1992 10% bloom stage, 1992 6070** 190NS 1220** 1300** 350NS 290NS 130NS 590** 90NS 60NS 180NS 95 3270** 180NS 1NS 1530** 40NS 960** 8NS 70NS 110NS 220NS 90NS 130 46.7 45.6 61.8 44.2 3.40 48.4 46.0 51.1 40.0 45.8 46.0 49.4 39.5 2.19 10% bloom stage, 1993 6270** 180NS 40NS 530** 130NS 1NS 50NS 210NS 60NS 30NS 70NS 100 53.7 54.6 55.8 39.6 Cultivar (Cv) Apica Barrier Apollo II WL316 Scarification (S) No 3-trifoliate leaf stage, 1992 64.4 60.4 56.2 46.2 2.97 Yes 59.5 49.9 59.8 41.9 Mean 62.0 55.2 58.0 44.0 2.10

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*, ** Significant at the 0.05 and 0.01 probability levels respectively; NS, not significant.

Table 3. Effect of seeding date on plant density expressed as a percentage of viable seed sown in 1992 Seeding date Growth stage 3-trifoliate leaf, 1992 10% bloom, 1992 10% bloom, 1993 30 April 72 63 62 10 June 47 46 32 30 July 45 42 42 SE 2.1 2.3 2.1

increased the plant density of high hard seed cultivars by about 17% but decreased the plant density of low hard seed cultivars by 10% (significant seed type scarification interaction; Table 2). The plant density at the three-trifoliate-leaf stage declined with the trial planting date (Table 3). At the 10% bloom stage and in the spring of 1993 the plant density was still highest for the earliest planting date. Modified Germination Tests In the first laboratory experiment, the effect of temperature regime on the germination of hard seed differed for the two

cultivars (P < 0.001) though the patterns were similar (Table 4). The highest rates of germination after 6 wk occurred with alternating 35/5C. The germination rate for constant 35C was almost as high but 11% of Barrier seedlings died. The percentage of germinated seed was similar at constant 20C and 5C. Alternating 20/10C killed most germinating seedlings. During May, June, and August of 1992, when the field trials were seeded, the average daily maximum air temperature ranged from 22.6 to 28.9C and the average minimum air temperature ranged from 8.8 to 14.2C. The temperatures chosen for study in the second germination experiment were: 1) the standard constant temperature (20C), 2) a temperature regime similar to air temperatures in the field (28/14C) and 3) the best regime from the first experiment (35/5C). The diurnal variation of the temperature in the soil where the seed is will be greater than in the air (Campbell 1977). However the second temperature regime represents a wide range of temperature and so should be a reasonable representation of at least some field conditions. More hard seed germinated in the first 4 wk of the second laboratory experiment than in the second 4 wk (Table 5).

HALL ET AL. ALFALFA GERMINATION AND ESTABLISHMENT

Table 4. Effect of temperature and cultivar on germination of hard alfalfa seed on moist blotting paper for 6 wk Cultivar Apica Temperature (C) 35/5 35 20 5 20/10 35/5 35 20 5 20/10 error. Germinated (%) 532.9z 362.8 131.9 152.0 10.6 682.7 272.7 232.5 182.6 20.8 Ungerminated Hard (%) 462.9 632.8 871.9 852.0 812.3 312.7 622.9 762.5 822.6 612.8 Dead, abnormal or fresh (%) 10.6 10.6 00.1 00.1 182.2 10.6 111.9 10.6 00.1 372.8

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Number tested 303 294 299 320 300 299 275 291 215 301

Barrier

zStandard

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After both 4 and 8 wk, there was a significant interaction (P = 0.03) among lighting, medium and temperature but the effect of the temperature regime predominated (P < 0.0001). The highest rates of germination again occurred at 35/5C. The germination rate was lowest at constant 20C and intermediate at 28/14C. The lighting and medium made little difference at constant 20C at either time or at 35/5C after 4 wk. After 8 wk at 35/5C the percent germination was highest on blotter under shade. After 4 wk at 28/14C, the percent germination on blotter under light was significantly higher than on the other two treatments. But after 8 wk at 28/14C the percent germination in soil under light was significantly lower than on the other two treatments. DISCUSSION The rate of germination of hard alfalfa seeds was increased by higher temperature during incubation (Tables 4 and 5). This effect of temperature was more pronounced when the higher temperature was present for only 8 h of the day. The effects of lighting and medium on the germination rate were much smaller than those of temperature. Thus, for cultivars having a high percentage of hard seed, the standard 7-d germination test which uses a temperature of 20C may underestimate the germination and consequent plant density of unscarified seed sown during spring or summer relative to that of scarified seed (Table 2) (Hall et al. 1993). The discrepancy between laboratory and field measurements of the effect of scarification reported by Hall et al. (1993) may be partly explained by the relationship between sowing rate and plant density. Using their data (Hall et al. 1993, Table 2, count 3) the relationship between plant density (D, plants m2) and sowing rate (R, kg ha1) was D = 19.9 + 28.6R. Thompson and Stout (1996) found a similar equation for irrigated alfalfa at 10% bloom. No significant non-linearity was detected in either case. The effect of a change in sowing (or germination) rate (dR) on the change in plant density (dD) determined from this equation is dD/D = dR/(R + 0.69). Averaging over the three sowing rates (1, 5, 10 kg ha1) used by Hall et al. (1993), this predicts that the 86% change in germination rate, observed in the laboratory would result in a 69% change in plant density in the field. There remains a residual discrepancy between this predicted 69% change and 40% change that Hall et al. (1993) observed.

Table 5. Percent germination of hard alfalfa seed (cv. Apica) as affected by lighting, medium and temperature regime. Sample sizes in parentheses Lighting Dark Shade Light Light Medium blotter blotter blotter soil 20C 28/14C 35/5C 472.5 (396) 442.5 (400) 452.0 (600) 4 weeks 71.0z (699) 182.7 (198) 51.5 (203) 232.1 (400) 61.7 (200) 151.5 (600) 8 weeks Dark Shade Light Light
zStandard

blotter blotter blotter soil error.

111.2 71.8 71.8

253.1 262.2 161.5

622.4 512.5 512.0

In the present study, a 1520% increase in plant density was observed at the 10% bloom stage in the 1992 seeding year (Table 2) when Apica and Barrier seed were scarified. However, no difference was detected at the earlier three-trifoliate-leaf stage or at the 10% bloom stage in 1993. Using the germination data (Table 1) and the relationship dD/D = dR/(R + 0.69), the predicted increase at the 10% bloom stage was 47% for Apica, 40% for Barrier and less than 1% for Apollo II and WL316. Thus, consistent with previous observations (Hall et al. 1993), the effect of scarification on plant density of the two cultivars with high percentages of hard seed was lower than predicted from the laboratory germination test results. In the year of seeding, freeze-thaw scarified seed of cultivars with a low percentage of hard seed had a 10% lower plant density at 10% bloom than unscarified seed (Table 2). No difference was detected in the earlier three-trifoliate-leaf stage or at 10% bloom in 1993. Thus we cannot rule out some small negative effect of scarification on the quick-germinating portion of the seed. Nevertheless, for cultivars with high hard seed content the net effect of scarification should be positive. The 1520% advantage observed for the high hard seed cultivars is a net value which includes any negative effects of scarification on the quick-germinating portion of the seed.

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As well as being affected by the germination rate, field establishment is influenced by other environmental factors and inter-plant competition. Inter-plant competition is unlikely to be a factor in this study or in that of Hall et al. (1993) because the relationship between plant density and sowing rate was linear (Thompson and Stout 1996) over the range of sowing rates used. Differences in environmental factors such as seeding depth, moisture, nutrients and temperature may be reflected in the differences in plant density observed at the different planting dates (Table 3). However at individual planting dates, these environmental factors were constant so they cannot explain the differences among treatments. Hall et al. (1993) observed that the percent germination in a 7-d test and the percent establishment for unscarified Apica were similar. In agreement, the Apica in the present study had 65% germination and a plant density of 65% at the three-trifoliate stage. For Barrier, the corresponding figures were 68% germination and a plant density of 60%. However, for cultivars with less than 1% hard seed, the plant density averaged only 51%. It seems unlikely that 100% of quickgerminating seed from cultivars with a high hard seed content would produce seedlings, but only 51% of quick-germinating seed from cultivars with very little hard seed would. This suggests that a fraction of the hard seed germinates and produces plants under field conditions. The results of the modified germination tests at field temperatures also support this view. Thus, the view that percent establishment is simply related to percent germination in the 7-d test is likely in error. To conclude, no changes in the germination rates of stored scarified seed were found over a 7-mo period. There may be some negative effect of scarification on the establishment of quick-germinating seed. However, for cultivars with a high hard seed content, the net effect of scarification was positive in the field. The rate of germination of hard seed was increased by higher temperatures during incubation in the laboratory. Thus some seeds identified as hard in a 7-d test at constant 20C are likely to germinate and contribute to an alfalfa stand grown in the field where temperatures are higher. A portion of hard seed in some lots may require scarification but not all hard seed require scarification to germinate.

ACKNOWLEDGEMENTS The authors acknowledge the technical assistance of Michelle Marginet, Darren Hines, Kim Dupuis and Carol Davidson.
Bass, L. N., Gunn, C. R., Hesterman, O. B. and Roos, E. E. 1988. Seed physiology, seedling performance and seed sprouting. Pages 961983 in A. A. Hanson et al., eds. Alfalfa and alfalfa improvement. Agronomy monogr. no. 29. ASA, Madison, WI. Bishop, Y. M. M., Fienberg, S. E. and Holland, P. W. 1975. Discrete multivariate analysis: Theory and practice. The MIT Press, Cambridge, MA. Campbell, G. S. 1977. An introduction to environmental biophysics. Springer-Verlag, New York, NY. Canada Department of Agriculture. 1979. Methods and procedures for testing seed. Laboratory Services Division, Food Production and Marketing Branch, Ottawa, ON. Cooper, C. S. 1977. Growth of the legume seedling. Adv. Agron. 29: 119139. Environment Canada. 1982. Canadian climate normals 19511980. Vol 2 Temperature. Vol. 3 Precipitation. Atmospheric Environment Service, Ottawa, ON. Hall, J. W., Stout, D. G. and Brooke, B. M. 1993. Hard seed and field establishment of irrigated alfalfa. Crop Sci. 33: 10251028. Leggatt, C. W. 1927. The agricultural value of hard seeds of alfalfa and sweet clover under Alberta conditions. Sci. Agric. 8: 243266. Leggatt, C. W. 1930. Further studies on the hard seed problem: alfalfa and sweet clover. Sci. Agric. 11: 418427. SAS Institute, Inc. 1990. SAS/STAT users guide, Version 6, 4th ed. SAS Institute, Inc., Cary, NC. Smith D. 1961. Note on the growth characteristics of alfalfa and red clover plants derived from hard seeds and seeds of different sizes. Can. J. Plant Sci. 41: 675678. Stout, D. G. 1990. Effect of freeze-thaw cycles on hard-seededness of alfalfa. J. Seed Technol. 14: 4755. Thompson, D. J. and Stout, D. G. 1996. Influence of sowing rate on dry matter yield, plant density and survival of lucerne (Medicago sativa) under dryland and irrigated conditions. J. Agric. Sci. (Camb.) 126: 301306. van Ryswyk, A. L., Stout, D. G., Hogue, E. J., Hall, J. W. and Roddan, B. H. 1993. Soil properties associated with alfalfa winter survival at Kamloops, B.C. Can. J. Soil Sci. 73: 141146. Weihing, R. M. 1940. Field germination of alfalfa seed submitted for registration in Colorado and varying in hard seed content. J. Am. Soc. Agron. 32: 944949.