Allelopathic Inhibitors in Seeds of Hevacleum laciniatum

Biological inhibitors present in the dry seeds of Heracleum laciniatum Horn have been studied by means of thin layer chromatography and various bioassays. The ethylacetate fraction of methanol extracts contained three main groups of inhibitors having the following R values when chromatographed on silica gel with chloroform: (1) 0.60–0.80; (2) 0.20–0.40; (3) 0.0–0.10. The group (1) inhibited lettuce seed germination and root growth, elongation of Avena first internode segments, growth of the fungus Cladosporium cucumerinum, and it caused leakage of betacyanin from red beet tuber slices. The groups (2) and (3) had no activity in red beet betacyanin leakage test, but they were inhibitory in the other bioassays used. The group (3) were generally less active than the group (2). The group (2) contained the furanocoumarins pimpinellin, bergapten, isobergapten and angelicin. Inhibitors present in the groups (1) and (3) have not been identified. Effects of five synthetic furanocoumarins (pimpinellin, bergapten, isobergapten, isopimpinellin, sphondin) on lettuce seed germination and Cladosporium growth were investigated. Pimpinellin was inhibitory to Cladosporium growth, and lettuce seed germination was inhibited by pimpinellin and sphondin.     

Germination and Dormancy in Immature and Fresh-mature Lettuce Seeds

The effects of red light, far-red light, Gibberellin A₃, andethephon were studied on the germination of lettuce seeds cv.Grand Rapids harvested at different stages of development. Seeds did not become capable of germination until 8 days afteranthesis. Red light promoted seed germination from the age of8–9 days following anthesis up to the newly mature stage.Ten or 11 days following anthesis, a large percentage of seedsbecame capable of germination in the dark and therefore couldbe considered not dormant. They were affected by far-red light,but less so than the mature seeds. The effect of light on the germination of developing seeds appearedto be similar to the known light effect on mature lettuce seedgermination. Gibberellin A₃ and ethephon had no effect on immatureand fresh seed germination. Lactuca sativa L., Lettuce, germination, dormancy, red light, far-red light, gibberellin A₃, ethephon     

Water Content and Phytochrome-induced Potential Germination Responses in Lettuce Seeds

Grand Rapids lettuce seeds (Lactuca sativa L.) with 6 to 8% water content show no light-induced germination responses, whereas in seeds with 15% or more water content, germination is promoted or retarded by red and far red light respectively. By adjusting seed water content, persistent potentiated responses to light are induced in the seeds at seed water levels much below that required for germination itself. Alternate moistening and drying of seeds in conjunction with red and far red irradiations show that potentiated responses may be phototransformed only with sufficient seed water. However, the process of drying and remoistening has no effect on potentiated responses.     

Allelopathy in Heracleum laciniatum: Inhibition of Lettuce Seed Germination and Root Growth

Both dark and red light germination of lettuce seeds (cv. “Maikönig”) as well as their root and hypocotol elongation were inhibited when the seeds were sown in petri dishes together with a few seeds of Heracleum laciniatum Horn. This inhibition was not significantly counteracted by the presence of gibberellic acid (GA₃) or/and 6-benzylaminopurine (BA). However, a large proportion of the lettuce seeds germinated abnormally (only cotyledons emerged) when treated with BA in the presence of Heracleum seeds. GA₃ had alone no significant effect on abnormal germination, but it counteracted the effect of BA to some extent. The inhibitory effect of Heracleum seeds gradually disappeared during a moist incubation period of one to seven days in darkness at 25°C. When lettuce seeds were pre-incubated together with Heracleum seeds for one to five days the remaining, non-germinated lettuce seeds had lost their ability for subsequent germination in darkness in distilled water. This induced dark dormancy was to a great extent broken by red light, but not by GA₃ or/and BA. H. laciniatum seeds inhibited the germination of Salix pentandra seeds and to some extent also the germination of radish but had no effect on the germination of spruce.     

Prehydration and Priming Treatments that Advance Germination also Increase the Rate of Deterioration of Lettuce Seeds

The influence of prehydration in water or priming in –1.5 MPa polyethylene glycol 8000 solution for various periods,followed by redrying, on germination rate and longevity of lettuce(Lactuca sativa L.) seeds (achenes) was determined during controlleddeterioration at 10% moisture content (fresh weight basis) and40°C. Short prehydration treatments (up to 1 h) had littleeffect on either germination rate or longevity, but significantlyimproved root growth rates. Increasing durations of prehydrationor priming reduced the mean time to germination by up to 61%relative to untreated seeds, but also reduced mean seed longevityby as much as 84% Prehydration and priming altered the relationshipsbetween germination rate and viability and between normal andabnormal seedlings during ageing. Prehydration in abscisic acidor at a temperature inhibitory to germination did not preventthe reduction in longevity under controlled deterioration conditions.While prehydration or priming treatments effectively acceleratesubsequent germination rates of lettuce seeds, the redried seedsare nonetheless highly susceptible to deterioration in storage. Key words: Lettuce, Lactuca sativa L., seed priming, seed deterioration, germination rate     

The Influence of Pre and Post-storage Hydration Treatments on Chromosomal Aberrations, Seedling Abnormalities, and Viability of Lettuce Seeds

A 2 h soaking treatment in distilled water, or in aqueous solutionsof cysteine, potassium iodide, or sodium thiosulphate, had nosignificant effect (P > 0.25) on the subsequent longevityof lettuce seeds (Lactuca sativa L.) in two different storageenvironments. Neither did these treatments influence relationsbetween loss in germination and the frequency of chromosomalaberrations observed during first mitoses after storage. Incontrast partial hydration of lettuce seeds after storage byexposure to moist air (humidification) or to an osmoticum (priming)reversed some of the damage which resulted from ageing. Mostof the benefits occurred during the first 3 d of humidificationduring which seed moisture content rose to 34 per cent, or duringthe first 7 d of priming when seed moisture content increasedto 44 per cent. Both post-storage hydration treatments reducedthe frequency of chromosomal aberrations, increased the rateof root growth, and decreased the frequency of morphologicallyabnormal seedlings. Either treatment could be of practical use,but it is suggested that humidification is more convenient.Consideration should be given to adopting a humidification treatmentas standard practice following long-term seed storage for geneticconservation. Lactuca sativa, lettuce, seed storage, seed viability, chemical pre-treatment, seed longevity, seed humidification (conditioning), seed priming, chromosome repair, seedling abnormalities     

Effect of malformin on phytochrome- and Ethrel-mediated responses

On etiolated cuttings of Phaseolus vulgaris malformin inhibitedseveral phytochromemediated responses. This included hypocotylhook-opening, leaf expansion, and inhibition of stem elongation.Malformin also inhibited anthocyanin synthesis by sorghum, buthad no effect upon lettuce seed germination in the light ordark. Malformin alleviated Ethrel-induced hook-retention, inhibitionof stem elongation, and root curvatures, but not Ethrel-inducedstimulation of lettuce seed germination in the dark. (Received February 19, 1975; )     

Some Aspects of the Reproductive Biology of Limonium vulgare Mill., and Limonium humile Mill.

The germination of two species of salt-marsh plants Limoniumvulgare Mill., and L. humile Mill., is investigated in relationto their ecology. Germination is inhibited by sea water, butgermination even in fresh water is low. Inhibition of germinationby sea water is negatively correlated with water uptake. However,a treatment in sea water stimulates subsequent germination infresh water. The effect of the sea-water treatment appears tobe one of osmotic shock which weakens the seed coat. Variationin response to the treatment is partly genetical and partlyenvironmental. Seeds in different parts of the salt-marsh willbe subject to differing conditions, both during seed developmentand during the interval between seed maturation and seed dispersal.In connection with this the effect of leaching by sea waterand fresh water is considered. The response to a sea-water treatmentvaries with time. Considered in the long term it shows somecyclical variation which might correspond with the tidal cycle.It is shown that seeds of Limonium can survive long periodsin sea water. There is a slow rate of germination during immersion.Most of those seeds which do not germinate in sea water surviveto germinate rapidly in fresh water. The response to oxygenconcentration, light, and temperature is also investigated.Experiments on seedling growth and establishment show the needfor fresh water and a suitable substrate. Mud gives a lowergermination but a higher survival, while sand gives a highergermination but a lower survival. The dispersal of Limoniumseeds in nature is investigated. Seeds of Limonium are mostabundant in the drift line, but even these only represent asmall fraction of the total produced. It is considered thatreproduction by seed is only important when a new area is beinginvaded. Subsequent reproduction is largely by vegetative spread.The ecological implications of the germination experiments arediscussed.     

Effects of Repetitive Acid Immersion, Red Light, and Gibberellin A3 Treatments on Phytochrome-mediated Germination Control in Skotodormant Lettuce Seeds

Dry lettuce seeds (Lactuca sativa L. cv. Grand Rapids), whichreceived 5 min far-red light (FR) 0.5 h after the onset of waterimbibition, showed 17% and 50% germination without and withacid immersion treatment (pH 0.1) for 1 h and rinsing with water,respectively. The acid treatment caused only 6% germinationor less in FR-treated seeds held for 10 to 30 d in dark storage.The 10 to 30 d skotodormant seeds did not respond to red light(R) or gibberellin A₃ (GA₃) singly, but showed 84% or higherpercentage germination if 1 h acid immersion was given beforeR or GA₃. The 20 d skotodormant seeds, which received R treatmentat day 10 but remained dormant showed 89% germination with onlyacid treatment. Similar values were obtained with 30 d skotodormantseeds which received one or two R treatments at day 10 or 20,i.e. the only requirement for these R-treated dormant seedswas an acid immersion. This releases the skotodormancy and rendersthe seeds more sensitive to R or GA₃, but the skotodormancywas initiated again if no light or hormone treatments were givenimmediately. The repetitive R or GA₃ treatments, which did notcause skotodormant seeds to germinate, lessened the degree ofskotodormancy. The germination of these skotodormant seeds canonly be induced by the synergistic action of R and GA₃. In thisstudy, GA₃ caused higher germination percentages in R-treatedskotodormant seeds than R stimulated in GA3-treated seeds. Itis suggested that (i) repetitive R or Ga₃ treatments maintaina high endogenous level of the far-red-absorbing form of phytochrome(P_fr) and GA activity, respectively, (ii) the accumulated stableintermediates of phytochrome persist in fully-imbibed skotodormantseeds for up to 20 d, without phytochrome expressing its functionuntil the seeds are acidified and (iii) a model is formulatedto interpret the results of acidification, growth promotersand R effects on germination of light-sensitive lettuce seeds. Key words: Phytochrome, Latuca saliva, seed germination, dark reversion of phytochrome, gibberellin A₃, acidification, skotodormancy     

Interaction of Moisture Stress and Three Phenolic Compounds on Lettuce Seed Germination

No interactions between water stress and three phenolic acids(p-coumaric, caffeic and ferulic acids) on lettuce (Lactucasativa L. var. Grand Rapids) seed germination were found. Probitanalysis indicated that mechanisms of action of water stressand the phenolic inhibitors were similar. The relative effectivenessof the compounds was p-coumaric > ferulic > caffeic. Nointeraction was found between p-coumaric and ferulic acid, whereasantagonism was found between caffeic acid and each of the othertwo phenolic acids. Lactuca sativa L., lettuce, germination, phenolic compounds, moisture stress, allelopathy, seed     

Stimulation of lettuce seed germination by ethylene

Ethylene increased the germination of freshly imbibed lettuce (Lactuca sativa L. var. Grand Rapids) seeds. Seeds receiving either red or far-red light or darkness all showed a positive response to the gas. However, ethylene was apparently without effect on dormant seeds, those which failed to germinate after an initial red or far-red treatment. Carbon dioxide, which often acts as a competitive inhibitor of ethylene, failed to clearly reverse ethylene-enhanced seed germination. While light doubled ethylene production from the lettuce seeds, its effect was not mediated by the phytochrome system since both red and far-red light had a similar effect.     

The use of water and some inorganic salt solutions to advance sugar beet seed. I. Laboratory studies

The emergence of sugar beet seedlings is often slow or irregular and insufficient plants may be established for the crop to yield fully. Treating the seed prior to sowing, such that the subsequent germination percentage is not reduced but germination is more rapid and better synchronised should be beneficial. A series of laboratory experiments was made to investigate seed treatment procedures involving water and various inorganic salt solutions. Many treatment combinations were identified which gave faster germination without decreasing germination percentage and left the seed dry and intact and suitable for sowing with conventional precision drills. No treatment was found which ‘primed’ the sugar beet seed and the term seed ‘advancement’ describes, more accurately, the effects observed. There was little difference in the performance of seeds optimally treated with water or some salt solutions generating an osmotic potential of between about –10 and –20 bars. However, the use of salt solutions, although more complicated, was preferred to water as inadvertent germination during treatment was less likely. The optimum treatment for the one bulk of sugar beet seed studied was, firstly, to wash the seeds for 3.5 h with water, then after air-drying, to moisten them on filter paper dampened with –15 bar (0–34 M) sodium chloride solution for 6 days at 15^oC followed by a final wash and air-drying.     

SEED GERMINATION OF AMERICAN POKEWEED (PHYTOLACCA AMERICANA). I. LABORATORY TECHNIQUES AND AUTOTOXICITY

Seeds of pokeweed (Phytolacca americana L.), an underinvestigated species, were studied to determine optimal conditions for laboratory germination. Soaking seeds in acid solutions prior to sowing increased rates of germination. Concentrated (conc) H₂SO₄ was more effective than conc NO₃ or conc HCl acids. Physiological evidence from seed germination studies suggests that autotoxicity, or intraspecific inhibition, exists in pokeweed, a species known to possess several biologically active compounds. Seed germination was investigated in the laboratory with aqueous extracts of vegetative and reproductive structures of the plant. The presence of extracts from most plant parts correlated with reduced or no germination by seeds of its own species, whereas the presence of distilled water correlated with high percentages of germination by control seeds. Whether diluted with water by 5-fold (20% v/v) or undiluted, juice of pokeweed fruits completely inhibited the laboratory germination of pokeweed seeds. Also, extracts of freshly harvested mature leaves, stems, and immature fruits inhibited seed germination. However, results with root extracts, obtained from a single individual, depended more on concentration, since the highest concentration (50% v/v) inhibited germination, and lower concentrations (10 and 20% v/v) increased germination percentages over control samples. Results with extracts of juvenile leaves correlated with neither inhibition nor promotion of germination. Thus, except for juvenile leaves and the root, most extracts of the pokeweed plant inhibited seed germination with more mature structures exerting more inhibition and less mature structures exerting less or no inhibition. The ecological implication of autotoxicity is that seeds are more dispersed through time and space. In regard to seed germination by other species, especially those taxa known to possess biologically active compounds, these and other results suggest that the phenomenon of autotoxicity might be more widespread than previously suspected.     

Germination activity of smoke residues in soils following a fire

A simple and rapid bioassay was implemented to detect the germination activity of extracts from soils in pre/post-burn conditions. Soil samples taken from burnt, unburnt and adjacent plots at depths of 0–2, 2–4, 4–6 and 6–8 cm before and after burning mesic grassland in South Africa were analysed for germination activity over an eight-week period. Soil samples were extracted using dichloromethane and bioassayed using Grand Rapids lettuce (Lactuca sativa L.) achenes (seeds). The Grand Rapids lettuce seeds exhibited greater germination percentages when treated with extracts from burnt soil compared to the other plots. The magnitude of the germination activity declined with time since the burn. The Grand Rapids lettuce seeds also exhibited significantly higher germination when treated with unburnt soil extracts compared to the control (distilled water) which indicates the existence of other factors controlling germination in unburnt soil. Germination activity in the adjacent plots decreased with time. These findings indicate that the germination activity of the smoke derived from burning plant-material diffuses into the soil and its persistence declines with time. Considering that the soil seed bank contains viable seeds, at a moderate depth, and that they are initially unaffected by the heat of the fire, then smoke residues following a fire can influence the germination and recruitment of plant species that are responsive to smoke-derived compounds and are represented in the germinable soil seed bank.     

Interaction between a plant-derived smoke extract,light and phytohormones on the germination of light-sensitive lettuce seeds

Plant-derived smoke extracts mimics the effect of red light on germination in light-sensitive lettuce seeds and partially overcomes the inhibitory effect of far-red light. Interaction between a smoke extract and gibberellins, cytokinins, abscisic acid and ethephon was investigated. Smoke acted synergistically with GA₃ and increased the sensitivity of the lettuce seeds to ABA. It seems likely that smoke affects membrane permeability or receptor sensitivity rather than influencing the phytochrome system of these seeds.Abbreviations R red light - FR far-red light - SM smoke extract     

Germination of Seeds in the Shadow of Plants

An attempt to prove the ecological significance of red-far red control mechanisms in seed germination was made. The seeds of 30 species were exposed beneath the plant canopies. All the normally light-stimulated seeds, and also seeds of 14 (out of 19) “insensitive’ species and seeds of 1 (out of 4) light-inhibited species, were inhibited or significantly retarded in their germination, as compared with seeds exposed to diffuse light in an artificial construction. Further experiments with “insensitive’ seeds of Lactuca sativa L. cv. Cud Vorburgu showed that after prolonged plant-shadow treatment the seeds became light-sensitive in the usual phytochrome-mediated manner. Seeds exposed under the plant canopies during a few days were extremely sensitive to red or white light, but this sensitivity diminished slowly in the course of treatment. The spectral composition of light filtered through the leaves shows great preponderance of far red radiation. The red-far red reversion can be simply obtained with the natural light and a leaf. In open stands bright weather retards considerably the germination of lettuce, cloudy weather brings about full germination. Some considerations on the ecological significance of seed behaviour, particularly as connected with plant competition, are given.     

Effects of Chemicals Often Regarded as Germination Stimulants on Seed Conditioning and Germination of Witchweed (Striga asiatica)

Gibberellin A₃ (GA₃), kinetin and zeatin promoted germinationof conditioned witchweed (Striga asiatica [L.] Kuntze) seedssimilar to dl-strigol and a natural stimulant from corn rootexudates. GA₃ promoted germination only with scarified seeds.However, these regulators, as well as dl-strigol and naturalstimulants, inhibited the conditioning of seeds and their subsequentgermination. On the other hand, abscisic acid induced 74% germinationof conditioned seeds but had no effect on seed conditioning.Other chemicals such as sucrose, NaNO₃, vitamin B₁, L-methionine,nicotinamide, indoleacetic acid and colchicine had no apparenteffect on seed conditioning, while sucrose, L-methionine andinositol induced about 30 % germination of conditioned seeds.Seeds generally required either a longer conditioning time ora higher terminal dl-strigol concentration to eliminate anyinhibition of seed conditioning induced by a compound. dl-Strigol, germination stimulation, parasitic plants, seed conditioning, seed germination, Striga asiatica (L.) Kuntze, weed control, growth regulating substances     

Selective inhibitors of germination of legume seeds in activated sludge compost

Water extracts of the compost produced from activated sludge and coffee residue were found to be selectively inhibitory to seed germination of some legumes. Germination rate of white clover (Trifolium repens L.), red clover (Trifolium pratense L.) and alfalfa (Medicago sativa L.) seeds were reduced to 2, 29 and 73% of the control, respectively, by water extracts of the compost (20 g l^–1). However, the extracts did not show any inhibition to seed germination of sorghum (Sorghum bicolor Moench), African millet (Eleusine coracana Gaertn.), and Komatsuna (Brassica rapa L.) at the same concentration. The inhibitors in the compost extracts were separated by ion-exchange chromatography and reverse-phase high performance liquid chromatography (HPLC) and the inhibitory activities of seed germination were tested with white clover seeds. Five inhibitors were isolated and identified as 3,4-dichlorophenylacetic acid (3,4-DCP), 3,4-dichlorobenzoic acid (3,4-DCB), 3,4,5-trichlorophenylacetic acid, 3,4,5-trichlorobenzoic acid and mono-2-ethylhexylphthalate by ¹H-, ¹³C-NMR spectroscopy and mass spectrometry. The inhibitory activities of some authentic chemicals of the inhibitors and the related compounds were compared. The results indicated that the main inhibitor in the compost could be 3,4-DCB, which was contained at the concentration of 6.58 mg kg^–1 compost and showed the strongest inhibitory effect on seed germination of white clover among the tested compounds.     

Seed dormancy in Acer: Endogenous germination inhibitors and dormancy in Acer pseudoplatanus L.

Summary Dormant seeds of Acer pseudoplatanus L. contain two zones of inhibition on paper chromatograms in 10:1:1 as detected by the lettuce and cress seed germination, and the wheat coleoptile bioassays. One zone at R_f 0.6–0.8 was partitioned into ethyl acetate at acid pH and was shown to contain ABA by its behaviour on GLC and isomerization under ultra-violet light. The other zone at R_f 0.9 was detected only in the germination bioassays and was partitioned into ethyl acetate over a range of pH indicating the presence of one or more neutral compounds.The inhibitors present in the embryo of dormant sycamore seeds inhibited the germination of non-dormant sycamore seeds at relatively low concentrations. A comparison with the effects of application of exogenous ABA indicated that endogenous ABA could not solely account for the inhibitory activity of seed extracts, which appeared to be due partly to the presence of ABA and partly to that of neutral compounds present in the embryo. Leaching treatments that removed dormancy led to a decrease in the level of inhibitors present mainly in the basic fraction. The exogenous application of kinetin to dormant sycamore seeds increased germination whereas gibberellic acid had no effect. Similar responses were obtained with lettuce seeds inhibited by the basic fraction of dormant sycamore seeds.It is suggested that an inhibitor-cytokinin interaction may be involved in the dormancy of sycamore seeds.     

Breaking of Seed Dormancy by Nitrate as a Gap Detection Mechanism

Germination of Planlago lanceolata seeds buried in a chalk grasslandwas higher in bare soil than in vegetated soil, and measurementof soil nitrate concentrations showed that they were high enoughto account for this stimulation. When seeds of P. lanceolatawere sown in pots of soil with or without plants, and wateredwith nutrient solution containing either no nitrate, or 14 mMnitrate (sufficiently high that not all nitrate was absorbedby the plants), the presence of plants inhibited germinationonly when the nutrient solution contained no nitrate. It wasconcluded that breaking of seed dormancy by nitrate can functionas a gap detection mechanism if nitrate concentrations in baresoil are high enough to break seed dormancy, but are too lowto break dormancy when vegetation is present Plantago lanceolata, seed, dormancy, germination, nitrate, gap detection