Germination response of four pasture species to temperature,light, and post-harvest period

Abstract

The purpose of this research was to explore the effect of temperature, light, and post-harvest period, and their interactions, on seed germination ecology of four common pasture species in the Mediterranean environment. Mature seeds of Diplotaxis erucoides, Hirschfeldia incana, Hyoseris scabra (Mediterranean distribution) and Sonchus oleraceus (cosmopolitan distribution) were subjected to seven constant temperatures (10–40°C, at intervals of 5°C) under continuous darkness, or a 12 h/12 h light/dark photoperiod at 30, 150 and 270 days after harvest (DAH). Cumulative germination and germination speed were determined. In all the tested species, except S. oleraceus, light significantly enhanced germination. S. oleraceus seeds maintained germination values over 90%, in a wide range of temperatures (10–35°C), in the dark as well as in light. Seeds of H. incana germinated well soon after seed dispersal. In D. erucoides and H. scabra, germination increased with storage period, while in S. oleraceus there was no effect of seed age. In all the species, moreover, no germination was recorded at 40°C. Temperature, light, and post- harvest requirement may be regarded as an adaptation strategy to ensure optimal conditions for seedling development and survival in Mediterranean species, while the species with a cosmopolitan distribution germinates under almost all tested conditions.     

Salinity and Temperature Effects on Germination for Three Salt-resistant Euhalophytes, Halostachys caspica, Kalidium foliatum and Halocnemum strobilaceum

The effects of three salinities (0, 100 and 500 mM NaCl) and four constant temperatures (10, 20, 30 and 35 °C) on seed germination of Halostachys caspica (M. B.) C. A. Mey., Kalidium foliatum (Pall.) Mop. and Halocnemum strobilaceum (Pall.) Bieb. were investigated. After seeds were treated with different concentrations of NaCl at constant temperatures of 10–35 °C for 16 days, ungerminated seeds were transferred to distilled water for 10 days to investigate the total germination; after this time, the ungerminated seeds from the 10 and 20 °C treatments were then moved to 35 °C for another 5 days to determine the final germination. The three plant species in the present experiment are salt-resistant euhalophytes growing in high saline soils in the Zhungur Basin in Xinjiang, a northwest province of China.Compared with germination under control conditions, germination percentages of all three species were not affected by 100 mM NaCl at 10–35 °C, while severely inhibited by 500 mM NaCl; germination percentages were very low at 10 °C up to 100 mM NaCl for all species; the optimum temperature for germination of H. caspica and K. foliatum was 20–30 °C, while 35 °C for H. strobilaceum, up to 100 mM NaCl; seeds did not suffer ion toxicity for all species, as evidenced by the high total germination after ungerminated seeds pretreated with 500 mM NaCl were transferred to distilled water at constant temperatures of 10–35 °C for 10 days, and the high final germination after the ungerminated seeds from the 10 and 20 °C treatments were subsequently moved to 35 °C for another 5 days; Halostachys caspica had greater sensitivity to increasing temperatures from 10 and 20 °C to 35 °C compared with the other two species.     

Combinational dormancy in seeds of Sicyos angulatus (Cucurbitaceae,tribe Sicyeae)

Seeds with a water‐impermeable seed coat and a physiologically dormant embryo are classified as having combinational dormancy. Seeds of Sicyos angulatus (burcucumber) have been clearly shown to have a water‐impermeable seed coat (physical dormancy [PY]). The primary aim of the present study was to confirm (or not) that physiological dormancy (PD) is also present in seeds of S. angulatus. The highest germination of scarified fresh (38%) and 3‐month dry‐stored (36%) seeds occurred at 35/20°C. The rate (speed) of germination was faster in scarified dry‐stored seeds than in scarified fresh seeds. Removal of the seed coat, but leaving the membrane surrounding the embryo intact, increased germination of both fresh and dry‐stored seeds to > 85% at 35/20°C. Germination (80–100%) of excised embryos (both seed coat and membrane removed) occurred at 15/6, 25/15 and 35/20°C and reached 95–100% after 4 days of incubation at 25/15 and 35/20°C. Dry storage (after‐ripening) caused an increase in the germination percentage of scarified and of decoated seeds at 25/15°C and in both germination percentage and rate of excised embryos at 15/6°C. Eight weeks of cold stratification resulted in a significant increase in the germination of scarified seeds at 25/15 and 35/20°C and of decoated seeds at 15/6 and 25/15°C. Based on the results of our study and on information reported in the literature, we conclude that seeds of S. angulatus not only have PY, but also non‐deep PD, that is, combinational dormancy (PY + PD).     

Effect of Temperature Regimes on Germination of Dimorphic Seeds of Atriplex prostrata

Dimorphic seeds of Atriplex prostrata were removed from cold dry storage monthly over a one year period to test for fluctuations in seed dormancy and germination rate. For each seed type, four replicates of 25 seeds were exposed to four alternating night/day temperature regimes mimicking seasonal fluctuations in Ohio: 5/15 °C; 5/25 °C; 15/25 °C and 20/35 °C with a corresponding 12-h photoperiod (20 μmol m⁻² s⁻¹; 400 – 700 nm). We found a significant three-way interaction of seed size, temperature and month for both percent germination and the rate of germination. Large seeds showed the greatest germination at the 20/35 °C and 5/25 °C temperature regimes and small seeds at the 5/25 °C regime. Large seeds had greater germination at all temperatures as compared to small seeds. Large seeds had the fastest germination rates at 20/35 °C followed by 5/25 °C whereas small seeds had the fastest rates at 5/25 °C followed by 20/35 °C. This revised version was published online in July 2006 with corrections to the Cover Date.     

‘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.     

Effect of Temperatures on Dormancy and Germination in Three Species in the Lamiaceae Occurring in Northern Wetlands

In the temperate region temperature is the main factor influencing the germination period of plant species. The purpose of this study was to examine effects of constant and fluctuating temperatures on dormancy and germination under laboratory and field conditions in the three wetland species Lycopus europaeus, Mentha aquatica and Stachys palustris. The results should give indications if the temperature-dependent regulation of dormancy and germination is phylogenetically constrained. Tests for germination requirements showed a minimum temperature for germination of 9 °C in Mentha and 12 °C in Lycopus and Stachys, and a maximum temperature of 33 °C for Lycopus and 36 °C for Mentha and Stachys. Fluctuating temperatures promoted germination in all three species but the amplitude required for high germination (>50%) differed: it was 8 °C in Mentha, 10 °C in Stachys and 14 °C in Lycopus (mean temperature 22 °C). The effect of temperatures on the level of dormancy was examined in the laboratory by imbibing seeds at temperatures between 3 °C and 18 °C for periods between 2 and 28 weeks, as well as by a 30-month burial period, followed by germination tests at various temperatures, in light and darkness. In the laboratory only low temperatures (≤12 °C) relieved primary dormancy in seeds of Lycopus, while in Mentha and Stachys also higher temperatures lead to an increase of germination. Dormancy was only induced in Lycopus seeds after prolonged imbibition at 12 °C in the laboratory. Buried seeds of all species exhibited annual dormancy cycles with lower germination in summer and higher germination from autumn to spring. Exhumed seeds, however, showed considerable differences in periods of germination success. Dormancy was relieved when ambient temperatures were below 12 °C. Ambient temperatures that caused an induction of dormancy varied depending on species and test condition, but even low temperatures (8 °C) were effective. At high test temperatures (25 °C) in light, exhumed seeds of all three species showed high germination throughout the year. The three species showed various differences in the effects of temperatures on dormancy and germination. Similarities in dormancy and germination found among the species are in common with other spring-germinating species occurring in wetlands, so it seems that the temperature dependent regulation of dormancy and germination are related to habitat and not to phylogenetic relatedness.     

Imbibition phases and germination response of Mimosa bimucronata (Fabaceae: Mimosoideae) to water submersion

Mimosa bimucronata is a pioneering tree that occurs predominantly in moist lowlands, floodplains and on margins of rivers and lakes in Latin America. The effect of submergence on seed germination in M. bimucronata was firstly studied. Patterns of water absorption by M. bimucronata seeds were investigated thereafter to assess the imbibition phases of scarified and unscarified seeds. The germination percentage was significantly higher in scarified than in unscarified seeds, and the velocity of seed germination also increased considerably in scarified seeds. Submergence duration did not significantly affect germination percentages of scarified and unscarified seeds. Therefore, seed viability after submersion suggests that M. bimucronata may display hydrochorous dispersal and also that seeds are able to germinate successfully in areas with frequent seasonal flooding. With respect to imbibition phases, phase II was very short or even absent for scarified and unscarified seeds; therefore, a plateau, where water absorption by seeds is established, was not observed. Finally, we verified that the passage from phase I to III was very tenuous and took a long time in seeds without scarification.     

A butenolide, isolated from smoke, can overcome the detrimental effects of extreme temperatures during tomato seed germination

The butenolide, 3-methyl-2H-furo[2, 3-c]pyran-2-one, is an highly active compound isolated from plant-derived smoke. This compound is known to stimulate seed germination in a wide range of plants akin to smoke or aqueous extracts of smoke. The present study attempted to elucidate the role of the butenolide in overcoming detrimental effects of low and high temperatures on tomato seed germination and seedling growth. The germination percentage followed a parabolic curve for temperatures ranging from 10 to 40°C, with 25°C being the optimum for all treatments. Control seeds showed radicle emergence at two extreme temperatures (10 and 40°C) and seedlings failed to develop further, even upon prolonged incubation. By comparison the butenolide-treated seeds grew into phenotypically normal seedlings at these non-optimum temperatures. The smoke–water-treated seeds had an intermediate response as only a fraction of germinated seed developed into normal seedlings. Seedling vigour indices as well as seedling weight were significantly higher (p ≤ 0.05) for butenolide-treated seeds at all temperatures. Furthermore, seedlings developed in the presence of the butenolide had about a 1:1 correspondence between root and shoot length. Butenolide-treated seeds grew better than the control seeds in the temperature shift experiments. A gradual decline in the vigour index values was recorded with an increased duration of incubation at the extreme temperatures. Results of the present study are very important from an horticultural point of view as they indicate the potential use of the butenolide compound in restoring normal seed germination and seedling establishment in tomato below and above optimum temperatures.     

Protocols for Use of Potamogeton perfoliatus and Ruppia maritima Seeds in Large‐Scale Restoration

Restoration of submerged aquatic vegetation from seed has been hampered by a lack of information on the appropriate conditions for collecting, processing, and storing seeds prior to dispersal. Seeds must be processed and stored under conditions that maintain seed viability, meet dormancy requirements, and prevent premature germination. This study examined the effects of collection date, processing technique, aeration, storage and induction temperature and salinity, and storage period on seed germination of two mesohaline aquatic species, Potamogeton perfoliatus and Ruppia maritima. Collection date and processing technique were significant factors affecting seed yield from donor populations. Seeds of both species remained viable and germinated best when stored at 4°C, and then exposed to freshwater induction conditions. However, their responses to other factors differed. Aeration during storage was necessary in order to maintain viability of P. perfoliatus seeds, whereas it was unnecessary for R. maritima seeds. Storage in freshwater at 4°C prevented germination of P. perfoliatus seeds, while high salinity during cold storage was necessary to minimize premature germination of R. maritima. Mean germination time of P. perfoliatus was dependent on storage salinity; in contrast, mean germination time of R. maritima seeds was dependent on induction salinity. These differences indicate that the methods required to produce large quantities of underwater plant seed amenable to large‐scale restoration efforts must be tailored to the specific requirements of individual species and must consider the range of processes from initial harvest through seed testing prior to field establishment.     

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.     

SEEDS OF KOSTELETZKYA VIRGINICA (MALVACEAE): THEIR STRUCTURE,GERMINATION, AND SALT TOLERANCE. I. SEED STRUCTURE AND GERMINATION

Dormancy of Kosteletzkya virginica (L.) Presl. seeds is primarily due to the impermeability of the seed coat to water. The impermeable structure is assumed to be, in other Malvaceae, the palisade layer of the seed coat. The percentage of seeds capable of imbibition and germination increased with increasing time of storage at low temperatures, but the release from dormancy was not accompanied by decreased seed coat resistance to pressure. Under natural conditions, mechanical damage to the seed coat due to changes in temperature and/or abrasion may render the seeds water permeable. It is not clear what causes water permeability during storage under laboratory conditions. During seed maturation and drying, the inner epidermis of the tegmen partly separates from the rest of the seed coat and an air space, which makes the seed buoyant, is formed around the region of the chalazal cleft. The optimal temperature for germination of K. virginica seeds is between 28 and 30 C in light or darkness.     

Seed germination ecology of the threatened endemic Iberian <Emphasis Type="Italic">Delphinium fissum</Emphasis> subsp. <Emphasis Type="Italic">sordidum</Emphasis> (Ranunculaceae)

Seeds of Delphinium fissum subsp. sordidum are physiologically dormant at maturity, with underdeveloped embryos; thus they have morphophysiological dormancy (MPD). The aims of this study were to determine the requirements for embryo growth, dormancy break and germination, to characterise the type of seed dormancy and to evaluate the effects of light, seed age, pollination mechanism, and inter-annual and inter-population variability on germinative ability. After 3 months of incubation at 5°C (cold stratification) in darkness conditions, the mean embryo length increased from 5.6 to 2.07 mm, with 76% of seeds germinating. Conversely, embryos of seeds incubated during 3 months at 20/7 or 28/14°C hardly grew and no germination was recorded. Since cold stratification was the only requirement for the loss of MPD, and both dry storage in laboratory conditions and warm stratification prior to cold stratification shortened the cold stratification period required for germination, it could be concluded that D. fissum subsp. sordidum seeds have intermediate complex MPD. Cold stratification and incubation in darkness conditions promoted higher germination percentages than those in light. In addition, germinative ability increased with seed age up to 8 months (reaching 96% at 5°C in darkness), showed a pronounced inter-annual and inter-population variability, as well as a significant decrease in seeds coming from pollination by geitonogamy. High temperatures (25/10 or 28/14°C) induced seeds to secondary dormancy, so seedling emergence in the greenhouse was restricted to February–March. The requirements for dormancy break and germination reflect an adaptation to trigger germination in late winter. This study is the first one to document a gradual increase in germination percentage with seed age for plant species with intermediate complex MPD.     

Environmental cues for germination of the invasive bunch grass <Emphasis Type="Italic">Pennisetum ciliare</Emphasis> (L.) Link

Responses of seed germination to air temperature, water potential, light, and smoke were studied in the laboratory for seeds of the invasive bunch grass Pennisetum ciliare (L.) Link (syn. Cenchrus ciliare L.; buffel grass). First introduced to North America during the mid-twentieth Century for establishing pastures, this African bunch grass has become an invasive species of concern. Across all the experiments conducted, a low germination was observed for P. ciliare fascicles that never exceeded 30 % at 21 days after sowing. Optimal day/night air temperatures for germination, controlled with an environmental chamber, were 25/15 and 30/20 °C, while extreme temperatures of 15/5 and 45/35 °C inhibited germination. By sowing seeds of P. ciliare under different water potentials, created with aqueous solutions of polyethylene glycol, an optimum of ?0.03 MPa led to the highest germination, while no germination was observed at ?1.0 MPa. Monochromatic optical filters were utilized to germinate seeds under various wavelengths, of which red (650 nm) and far red (730 nm) led to the highest germination. In addition, seeds that were incubated in the dark had higher germination than those incubated under white light. Incubation in smoke water, which can stimulate germination of pyrophytic species, resulted in a marginal inhibition of germination compared with imbibition with distilled water.     

The germination characteristics of <Emphasis Type="Italic">Scrophularia marilandica</Emphasis> L. (Scrophulariaceae) seeds

Investigations on seeds of Scrophularia marilandica L. were undertaken to determine their germination requirements. Seeds were collected from three naturally occurring sites and one greenhouse-grown population in London, Ontario in September and October of 1997. Some were set to germinate immediately after collection; others were stored in or on soil outside and/or under controlled laboratory conditions before testing. Germination was assessed under two light/temperature regimes (35°C 14 h light, 20°C 10 h dark and 25°C 14 h light, 10°C 10 h dark), in continuous darkness, and in the presence of two germination-promoting chemicals (GA₃ and KNO₃). Fresh seeds germinated best at 35/20°C, while stored seeds germinated best at 25/10°C. No differences in percent germination were found among three seed-maturity stages. All chemical treatments, except 0.01 M KNO_3, increased percent germination. Significant differences were found both among and within sites for most chemical treatments, but exposure to 3 × 10⁻⁴ M GA₃ caused almost every seed to germinate. When compared to the control, both the gibberellic acid and the soil-storage treatments contributed to faster germination. Exposure of seeds to naturally prevailing conditions on the soil surface followed by testing under the 25/10°C regime produced the highest percent germination. No seeds germinated in the dark. In summary, seeds of S. marilandica exhibit physiological dormancy, which can be alleviated by exposure to light, after-ripening and/or cold stratification. It is probable that the differences in germination response among sites can be attributed to differences in environmental conditions during seed production. These experiments indicate that the seeds of S. marilandica must be buried shortly after dispersal in order to form a persistent seed bank.     

Effects of Temperature of Mother-plant Environment on Yield and Germination of Seeds of Lettuce (Lactuca sativa)

Experiments on the production of two separate crops of lettuceseeds, each in three different temperature environments, andsubsequent tests on the seed are described. Low production temperatures(20 °C day, 10 °C night) gave a low yield of large seeds,and high temperatures (30 °C, 20 °C) gave a higher yieldof much smaller seed; the highest yield came from medium temperatures(25 °C, 15 °C), which gave medium-sized seed. After-ripening,manifested as an increase in percentage germination at hightemperatures with increase in seed age, occurred in seed fromall three production environments of the first crop, thoughthere were differences in degree, and in that from the two higherproduction temperatures, but not the lowest, of the second crop.Measurements of the forces required to penetrate the layerssurrounding the embryo showed an inverse relationship with temperatureof the production environment for pericarps but not for endosperms,and a gradual reduction during storage for pericarps but notendosperms. Measurements of germination potential showed thatembryos from seeds produced in cool conditions were less ableto cope with high temperatures than those from warner conditions.These results are discussed in relation to the control of germinationin lettuce. Lettuce, Lactuca sativa (L.), seed production, germination, seed coverings, germination potential     

Artificial ageing of Vicia faba L. seeds caused by prolongation of G-1 phase

Broad bean (Vicia faba L. “Inovec”) seeds were artificially aged by means of storage at 30 %, resp. 25 % water content at 25 °C for 7-days to study the consequences on germination, root length and frequency of chromosomal aberrations. Under these conditions, significant changes in all parameters were observed. An increase of frequency of chromosomal aberrations in ana-telophase cells was confirmed by evaluation of c-metaphase cells. Synergic effect of artificial seed ageing was studied on different harvests of old seeds. Possible principles of this effect on cell level are discussed.     

Germination response of black and yellow seed coated canola (<Emphasis Type="Italic">Brassica napus</Emphasis>) lines to chemical treatments under cold temperature conditions

Seed quality is a key critical component to produce well established and vigorous seedlings under cool soil (<10°C) conditions experienced in Western Canada. A simple, relatively quick germination assay is required to separate small differences in seed germination which can have a significant impact on seedling growth. It has long been established that phytohormones regulate seed germination: abscisic acid inhibits germination whereas gibberellins enhance germination. We investigated the effects of ABA, GA, ethylene and inhibitors of these phytohormones alone and in combination on the germination rate of a black and a yellow seed canola (Brassica napus) imbibed at 8°C. The effects of either saline solutions, osmotic solutions, fusicoccin or testa on the germination of canola seeds imbibed at 8°C were also investigated. This temperature is representative of the soil temperatures experienced in the early spring of Western Canada. The two canola seed lines, especially the yellow seed line, were very sensitive to increasing concentration of saline solutions at 8°C, but not at 23°C; however, iso-osmotic solutions that reduced water potential were more inhibitory. The seed coat (testa) including the endosperm was a major factor affecting the germination rate of the yellow seed line at 8°C, however, GA₄₊₇ overcame the inhibitory effect of the testa, whereas ABA exacerbated it. Fusicoccin was more stimulatory to germination than GA₄₊₇, however, unlike GA₄₊₇, it was unable to overcome the inhibitory effect of paclobutrazol, a GA biosynthesis inhibitor. Fluridone, an ABA biosynthesis inhibitor, was unable to overcome the inhibitory effects of a saline solution suggesting that the inhibitory effect was not due to elevated ABA levels. Ethylene, a stimulator of germination did not appear to be involved in the germination of these two lines. Controlled deterioration at 35°C, 85% RH could be either partially or completely overcome by exogenous GA₄₊₇. This study demonstrated the effect of hormones, salinity and testa on the germination of canola seeds under less than ideal environmental conditions.     

Germination responses to temperature responsible for the seedling emergence seasonality ofPrimula sieboldii E. Morren in its natural habitat

Temperature requirements for the breaking of seed dormancy and germination inPrimula sieboldii E. Morren and the annual surface-soil temperature regime in one of its natural habitats were investigated in order to clarify the germination responses determining the seedling emergence seasonality of the species. In a grassland nature reserve in an abandoned flood plain of the Arakawa River, natural seedling emergence of the species was shown to be restricted to mid- to late-spring before the closure of seasonal vegetational gaps, when the daily mean soil surface temperature reached about 15°C, accompanied by large daily fluctuations of about 10°C. Mature seeds collected in late June were never able to germinate at any constant temperature in the range of 8–40°C unless they had been previously subjected to moist-chilling treatment. The proportion of seeds which were released from dormancy increased with increasing duration of the moist-chilling treatment at 2°C, 70–85% of seeds becoming germinable at 16–28°C after 12 weeks of pretreatment at 2°C. The thermal time required for the germination of the thus-pretreated seed population was 905–1690 Kh with a base temperature of around 5°C. Fluctuating temperatures between 24°C and 16 or 12°C had a remarkable dormancy-breaking effect, inducing considerably quick germination in most of the seeds previously subjected to 2°C moist-chilling for 8 weeks.     

Factors Affecting Attachment of Enterobacter cloacae to Germinating Cotton Seed

Abstract Attachment of Enterobacter cloacae EcCT-50,—a biological seed protectant used to control the seed-rotting fungi, Pythium ultimum—to cotton seed was examined using conventional fluorescent microscopy (CFM), scanning electron microscopy (SEM), and laser scanning microscopy (LSM). In sand microcosms, E. cloacae quickly attached to the seed coat, with maximum attachment, 3 to 5 h after inoculation at 24°C. In contrast, initial attachment of non-bacterized seed by Pythium ultimum was not observed until 6 h (and not until 8 h on bacterized seeds). Comparison of the movement of E. cloacae and P. ultimum in seed exudate gradient semi-soft agar showed faster movement by the bacterium within the first 6 h, and reduction of P. ultimum hyphal and germ tube growth in the presence of the bacterium. Microscopic observation of the seed coat revealed an early, intimate association, mediated, in part, by fimbriae, and confirmed a loose association of E. cloacae with the seed coat previously reported. Spatially, the attached E. cloacae cells were distributed over the entire surface of the seed coat, but were especially abundant in the groves and near cracks where water imbibition and seed exudate release may occur. As the seed germinated and exposed various seed tissues, the bacterium rapidly attached to these tissues. Attachment of the bacterium to the surface of intact germinating seeds, excised seed coat, polystyrene, and glass was 300, 110, 51, and <1 cell field⁻¹ 3 h⁻¹, respectively, suggesting that attachment is enhanced by seed germination. Attachment of E. cloacae to the seed coat was optimum in sands with high water concentrations, at temperatures of 18 to 30°C, and at times that corresponded with optimum water imbibition during germination. Using several assays, attachment was shown to be enhanced by seed exudate, and compounds such as methanol, fructose, and calcium. The results suggest that the release of certain nutrients and water imbibition during germination may play a role in the rapid attachment to the seed by E. cloacae. The ability of E. cloacae to rapidly move and attach to the seed coat may be related to its ability to function as a biocontrol agent. Received: 24 April 1997; Accepted 29 October 1997