Germination of Tagetes minuta L. I. Temperature Effects

Initial studies have indicated that Tagetes minuta achenes haveboth a temperature and a light requirement for germination.Temperatures tested were 10, 20, 25, 30 and 35 °C. Germinationwas optimal at 25 °C under white light conditions. Underthese conditions 100 per cent of achenes germinated within 7days of imbibition. There was no germination at 10 or 35 °Ceither in the light or in the dark. Achenes imbibed and incubatedat 35 °C for 4 days showed no visible signs of germinationbut on transfer to 25 °C, 100 per cent of these achenesgerminated within 24 h. Furthermore, achenes given this hightemperature (35 °C) treatment could be dried at 25 °C,re-imbibed at 25 °C and again 100 per cent of achenes germinatedwithin 24 h of re-imbibition. This rapid germination responsefollowing removal from the high temperature regime could alsobe induced by transfer to temperatures of 20 °C or 20 °C(16 h) alternating with 10 °C (8 h). Tagetes minuta L., weed seeds, germination, temperature, light     

Germination of Tagetes minuta L. II. Role of gibberellins

The effect of the application of gibberellins to Tagetes minuta L. achenes (seeds) was determined at both 25°C, the optimal germination temperature, and 35°C, at which temperature the achenes are thermoinhibited. Both GA₃ and GA₄₊₇ accelerated germination at 25°C. Seed germination at 25°C was inhibited by paclobutrazol, but on subsequent application of GA₄₊₇ rapid germination was induced. Following application of GA₃ or GA₄₊₇ to thermoinhibited seeds, a significantly higher final germination percentage was observed than in the distilled water control. However, endogenous gibberellin levels in germinating and thermoinhibited seeds did not differ significantly.     

Effects of Temperature and dl-Strigol on Seed Conditioning and Germination of Witchweed (Striga asiatica)

Seed conditioning and germination in witchweed (Striga asiatica(L.) Kuntze) were temperature-dependent. With higher conditioningtemperatures, shorter conditioning time was required for germinationwith terminal dl-strigol (strigol) treatment at 30 °C. Maximumgermination (80–100%) was obtained by conditioning inwater at 20, 25, 30 and 35 °C for 14, 7, 5 and 3 d, respectively,and terminally treating with 10^–6 M strigol at 30 °C.Seeds conditioned in 10^–8 M strigol instead of water germinatedmuch less with the same terminal strigol treatment. Generally,conditioning was slower when seeds were conditioned in strigolrather than water. The reduction in germination rate by pretreatmentin strigol or pretreatment at low temperatures could be overcomeby increasing the terminal strigol concentration in the germinationtest. Conditioned seeds did not germinate at 10 and 15 °Cwith a terminal 10^–6 M strigol treatment but yielded closeto maximum germination at 25, 30 and 35 °C with the sameterminal strigol treatment. To obtain maximum germination, boththe minimum conditioning temperature and the minimum germinationtemperature for conditioned seeds were 20 °C. Factors suchas conditioning time, and strigol concentration and temperatureduring conditioning and/or germination determine whether seedsremain in the conditioning phase or shift to a germination phase. dl-Strigol, germination stimulation, parasitic plants, seed conditioning, seed germination, Striga asiatica, temperature, weed control     

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

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

Changes in Membrane Fluidity and Protein Composition during Release of Cucumber Seeds from Dormancy by a Higher Temperature Shift

Dormancy of seeds of cucumber (Cucumis sativus L.) was inducedby imbibing in -1.8 MPa polyethylene glycol 6000 (PEG) solutionand pulsing with far red light for 15 min prior to washing anddrying. When re-imbibed with water at 20 °C, dormancy wasbroken by raising the temperature to 30 °C for 6 h. Thistreatment was also effective when -0.9 MPa PEG was present duringre-imbibition and high temperature. Seeds with broken dormancywere found to germinate in water over a smaller temperaturerange than seeds in which dormancy had not been induced. Whenthe duration of the temperature shift to 30 °C was varied,germination percentage increased from 7 to 60% after 6 h, butlonger exposures up to 12 h had no further promoting effect.The time course of germination after transfer to water following6 h at 30 °C in PEG showed piercing of the perisperm-endospermenvelope after 9–12 h and radicle protusion after 12–15h. If PEG was retained after high temperature treatment no visiblegermination was observed. Thus, to study membrane fluidity andthe protein content associated with germination, seeds weresampled 9 h after high temperature treatment. To study the germinablebut not germinating state, seed held in PEG for 9 h rather thanin water was used. Dormant seed was sampled before the hightemperature treatment. Membrane fluidity was assessed usingfluorescence polarization of membrane fractions treated withDPH (1,6-diphenyl-1,3,5-hexatriene) or its derivatives. Membraneproteins were compared using one-dimensional SDS-PAGE electrophoresis.Intracellular membrane fluidity was not increased in the transitionfrom the dormant to germinable state, but did increase in thetransition to germination. There were no detected changes inintracellular membrane proteins during either transition. Inplasma membrane fractions, fluidity increased during both transitions,while a marked increase in 21, 18 and 17 kD proteins was observedin the transition from germinable to germinating state. Thusmodification of plasma membrane fluidity rather than changesin protein profile is associated with the high temperature releaseof cucumber seeds from dormancy. Copyright 2000 Annals of BotanyCompany     

Osmoconditioning as a Means of Counteracting the Ageing of Pepper Seeds during High-temperature Storage

Sweet pepper seeds were osmotically conditioned in 0.4 M mannitolsolution for 4 d (at 25 °C, in darkness) before or afterstorage at 35 °C for up to six months, and their germinationand viability was compared with that of untreated seeds storedunder the same conditions. Seeds that had been osmoconditionedprior to storage retained a high rate of germination and germinatedto a high final percentage (from 80 to 50 per cent) at both15 and 25 °C throughout the storage period. By contrast,both the rate and total level of germination of untreated pepperseeds declined rapidly at both germination temperatures, andby three months of storage the total level of seed viabilitywas already less than 10 per cent. Seeds that were first storedat 35 °C, and then osmoconditioned just prior to germination,showed a decline in germinability which when tested at 25 °Cwas the same as for untreated seeds, while tested at 15 °Coccurred at a slightly slower rate than for untreated seeds. It is evident that osmoconditioning prior to storage, in additionto the acceleration of germination, resulted in a dramatic delayof the ageing rate, thus increasing considerably the longevityof seeds. On the other hand, osmoconditioning after storagedid not seem to have any significant effect on seed viability,though it enhanced the germination rate. Capsicum annuum, sweet pepper, seed, germination, osmoconditioning, priming, storage, viability, ageing, longevity     

Germination of Stored Cassava Seed at Constant and Alternating Temperatures

Cassava seed is only capable of germinating over a restrictedrange of constant temperatures. During storage the optimum constanttemperature for germination decreases from about 35 to 30 °Cor possibly less. The rate at which the optimum temperaturechanges during dry storage increases with increase in storagetemperature over the range 0 to 40 °C. Some alternating-temperatureregimes (16 h at the lower temperature; 8 h at the higher temperature)can provide conditions as favourable for germination as theoptimum constant temperatures. Furthermore, it has been shownthat temperature alternation itself is stimulatory because whenthe range of the alternation does not include the optimum constanttemperature value, percentage germination is often higher thancould be obtained at any constant temperature within the range,though this stimulatory response declines during storage. Forthese reasons it is provisionally recommended that cassava seedshould be germinated at 25/35 °C which is as stimulatorya treatment as any which has so far been investigated and hasthe advantage of encompassing the range over which the optimumconstant temperature changes during storage. Manihot esculenta Crantz, cassava, germination, dormancy, seed viability, storage of seeds, after-ripening     

Ethylene Involvement in Dormancy Release of Ricinodendron rautanenii Seeds

Ricinodendron rautanenii (manketti) seeds respond to as littleas 10^–2 µl I^–1 ethylene. Both dry and imbibedseeds respond to ethylene treatments. The length of the imbibitionperiod prior to the ethylene treatment is not important as seedswhich were simultaneously gassed and imbibed germinated successfully.Seeds were sensitive to the gas at temperatures between 25 and35 °C. No temperature effect was observed in seeds whichwere transferred from temperatures below 20 °C, or above35 °C, to 30 °C. There is no period of carbon dioxidesensitivity associated with the ethylene treatment and the effectsof the ethylene treatment persisted in both dry and imbibedseeds when subsequently maintained (desiccated) at room temperaturefor 8 d Ricinodendron rautanenii, manketti nut, ethylene, germination     

Development of Endopeptidase Activity in Cotyledons of Vigna mungo Seedlings: Effects of Exogenously Applied End-Products and Plant Hormones

The development of endopeptidase activity in cotyledons of Vignamungo seedlings was examined after application of exogenousamino acids, sugars and plant hormones. The endopeptidase activityin the cotyledons fell when germinating seeds were allowed toabsorb a solution of amino acids at high concentrations, andit was postulated that this effect might have been caused inpart by osmotic stress and in part by end-product repression.Protein immunoblotting with an antiserum against SH-EP, themajor cysteine endopeptidase occurring in the cotyledons, showedthat sugars and amino acids at high concentrations also delayedthe post-translational processing of SH-EP intermediates. Endopeptidaseactivity equivalent to nearly twice that in controls was observedwhen GA₃ was applied at 10 to 100 µM to cotyledons thathad been detached from the embryonic axis. In addition, naphthaleneaceticacid at 1 to 100 µM, kinetin at 1 to 10 µM and jasmonicacid at 1 to 10 µM also increased the activity to a limitedextent. Results of pulse-chase experiments suggested that theeffect of GA₁ on the endopeptidase activity in the detachedcotyledons was attributable to suppression of the degradationof the enzyme. Protein immunoblotting revealed the presenceof 34-kOa and 35-kDa intermediates of SH-EP in addition to previouslyreported 36-kDa and 43-kDa intermediates. (Received June 26, 1995; Accepted October 16, 1995)     

Lipase Activity in Germinating Seedlings of Cucumeropsis edulis

Acetone powders prepared from dormant and germinating seedlingsof Cucumeropsis edulis contained an active lipase system whichshowed increasing lipase activity with germination. Kineticproperties indicated maximum activity at pH 5 and at a temperatureof 37 ° C, and the enzyme exhibited a zero-order reactionrate for the first 2 h of hydrolysis. The results obtained showedthat the enzyme is non-specific in its mode of action, hydrolysingfatty acids irrespective of chain length, degree of unsaturation,and the position of fatty acids in the triglycerides. The deterioration and spoilage of ‘ground’ seedsin storage has been shown to be due to the release of endogenouslipase during grinding. The release of lipase is greater thegreater the water content of the seeds     

Storage Behaviour of Salix alba and Salix matsudana Seeds

Effects of dehydration, storage temperature and humidificationon germination of Salix alba andS. matsudana seeds were studied.Newly released seeds showed 100% germination before and afterdehydration to 11–12% moisture content. Germination ofthe high vigour lot (100% initial normal germination) was notaffected by dehydration to 6.7% moisture content but germinationdecreased with further dehydration to 4.3%. The lower vigourlot (75% initial normal germination) was more susceptible todehydration and germination decreased following dehydrationto 6.7% moisture content. Dry seeds of both species survivedimmersion in liquid nitrogen without loss of viability. Thegermination of seeds stored with 9% moisture content decreasedto 35–40% in 5 months at -20°C or in 2 months at 5°C.However, at 25°C seeds entirely lost viability within 2weeks. Seeds showed improved performance when stored at -70°C> - 20°C > 5°C > 25°C and tolerated dehydrationto a moisture content in equilibrium with 15% relative humidity.Results suggest that they are orthodox in storage behaviouralthough they are short-lived. Humidification treatment of lowvigour seed lots resulted in a remarkable increase in germinationpercentage. Copyright 2000 Annals of Botany Company Salix alba, Salix matsudana, willow, seed storage behaviour, dehydration, humidification, cryopreservation     

Characterization of a Small Sulphur-Rich Storage Albumin in Seeds of Alfalfa (Medicago sativa L.)

A 2S albumin fraction was characterized in seeds of alfalfa{^{Medicago sativa} L.). This low molecular weight (LMW) familyof disulphide-bonded proteins represents a major nitrogen andsulphur storage reserve for the alfalfa seed Characteristicof seed storage proteins, the 2S albumins are abundant in nitrogen-richglutarrune/glutamate/asparagine/aspartate (32%) In addition,this LMW fraction is high in cysteine (9%) and methionine (4%),amino acids which are under-represented in legume seed globulins.These 2S proteins start to accumulate during the early cotyledonstage of development, and are mobilized following germinationPulse-chase labelling experiments show that the 2S proteinsare synthesized as 'preproproteins', similar to 2S proteinsin other seeds. However, alfalfa 2S albumins are immunologicallyunrelated to these proteins. Key words: Seed development, sulphur-containing 2S storage protein, alfalfa (Medicago sativa)     

Osmoconditioning and Ageing of Pepper Seeds During Storage

The effects of osmoconditioning on the germination at 15 and25 °C of pepper (Capsicum annuum L.) seeds were studiedover a 3-year period with respect to temperature of storage.Untreated seeds stored at 5 °C showed high germinabilitythroughout the entire storage period, whereas untreated seedsstored at 25 °C showed a progressive decline in germinability,especially when assayed at 15 °C. Seeds that had been osmoconditionedprior to storage retained a high level of germinability irrespectiveof either storage or germination temperatures. When seeds thathad been stored at 25 °C were osmoconditioned after storage,there was a significantly higher germinability (assayed at 15 °C) in comparison with the corresponding untreated seeds.Seeds that were osmoconditioned twice (prior to and after storage)germinated in a similar way to those that had been osmoconditionedonce only Lactuca saliva L., lettuce, Hordeum oulgare L., barley, seed storage, moisture content, relative humidity, water potential, temperature, oxygen     

The Survival of Germinating Orthodox Seeds after Desiccation and Hermetic Storage

Seeds of barley (Hordeum vulgare L.) and mung bean (Vigna radiata(L.) Wilczek), with orthodox seed storage behaviour, were imbibedfor between 8 h and 96 h at 15 °C and 25 °C, respectively,while barley seeds were also maintained in moist aerated storageat 15 °C for 14 d. These seeds and seedlings, together withcontrols, were then dried to various moisture contents between3% and 16% (wet basis) and hermetically stored for six monthsat —20°C, 0°C or 15°C. In both species, neitherdesiccation nor subsequent hermetic storage of the control lotsresulted in loss in viability. The results for barley seedsimbibed for 24 h were similar to the control, but desiccationsensitivity increased progressively with duration of imbibitionbeyond 24 h in barley or 8 h in mung bean; these treatmentsalso reduced the longevity of the surviving seeds in air-drystorage. Loss in viability in barley imbibed for 48 h was mostrapid at the two extreme seed storage moisture contents of 3·6%and 14·3%, and in both these cases was more rapid at15 °C than at cooler temperatures. Similarly, for mung beanimbibed for 8 h, loss in viability was most rapid at the lowest(4·3%) moisture content, but in this case it was morerapid at –20 °C than at warmer temperatures. Thus,these results for the storage of previously imbibed orthodoxseeds conform with the main features of intermediate seed storagebehaviour Key words: Barley, Hordeum vulgare L., mung bean, Vigna radiata (L.) Wilczek, desiccation sensitivity, seed longevity, seed storage behaviour     

Dormancy in seeds of Phellodendron wilsonii is Mediated in Part by Abscisic Acid

A regime of temperatures alternating between 35°C (8 h)and 10°C (16 h) proved to be a specific method for breakingdormancy of seeds of Phellodendron wilsonii. The relationshipbetween the germination capacity after about 13 days of incubationunder this regime and the logarithm of the amount of ABA inthe seed before incubation yielded a high correlation coefficient.The final germination capacity under this regime was poorlycorrelated with the amount of ABA in seeds before incubation.These observations suggest that at least part of the germinationbehavior is controlled by the amount of abscisic acid in theseed. Stratification decreased the amount of ABA but resultedin only 8–25% germination relative to other dry storedseeds failed to germinate under suboptimal conditions, namely,incubation at 22°C with 12 h light. This phenomenon indicatesthat some unknown mechanism is the major factor that controlsthe dormant state, and the effect of this mechanism are overcomeonly by the specific alternating-temperature regime. Fluctuationsin temperature failed, however, to reduce the ABA content ofseeds during the incubation period. We conclude that some unknownfactor contributes to the dormancy of seeds of P. wilsonii althoughthe level of ABA definitely plays a minor role in the maintenanceof seed dormancy. (Received January 25, 1993; Accepted November 25, 1993)     

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.     

Seed Germination of a Halophytic Grass Aeluropus lagopoides

Aeluropus lagopoides(Linn.) Trin. Ex Thw. (Poaceae) is a perennialgrass distributed from coastal Sindh and Balochistan to salineflats of Punjab, Pakistan. Seeds collected from an inland populationofA. lagopoides located on the University of Karachi campuswere germinated under various levels of salinity (0, 100, 200,300, 400 and 500 m M NaCl) and temperature regimes (10/20, 15/25,20/30 and 25/35 °C) in a 12 h dark/12 h light photoperiod.Highest germination was obtained under non-saline conditions,and an increase in NaCl concentration progressively inhibitedgermination. Inhibition of germination was greater at coolertemperatures (10/20 °C) when no seed germinated above aconcentration of 300 m M NaCl. The germination response at moderatetemperatures (20/30 °C) was optimal, with 30% of seeds germinatingin 500 m M NaCl. The rate of germination decreased as salinityincreased. Germination rate was highest at 20/30 °C andlowest at 10/20 °C. Seeds were transferred from salt solutionsto distilled water after 20 d and those from high salinitiesrecovered quickly at warmer temperatures with an optimal responseat 20/30 °C. Copyright 2001 Annals of Botany Company Aeluropus lagopoides, germination, halophyte, Karachi, salinity, temperature     

Effects of Photoperiod on Germination of Seeds of Talinum triangulare (Jacq.) Willd

Seed germination in Talinum triangulare as affected by photoperiod,with or without previous incubation in the dark in water at25 or 4 °C, was studied. The time course and quantity ofseed germination in photoperiods of 1 h and above were similarwith or without dark pretreatment, but the time to half maximumgermination was reduced from 12 days in non-dark pretreatedseeds to 4 days in seeds given 20 days in the dark at 25°C.A photoperiod of 0·25 h gave a lower rate and total germinationthan photoperiods of 1 h and above. Un-pretreated seeds required17 cycles of 24 h photoperiod for maximum germination as comparedwith 7 or less cycles if the seeds received more than 10 daysdark pretreatment at 25 °C. Both the rate and total germinationin light increased as the length of dark pretreatment at 25°C was increased from zero to 30 days. Incubation of theseeds in water in the dark at 4 °C for 5 to 30 days priorto illumination at 21 °C, reduced both the rate and quantityof seed germination in light as compared with those similarlyincubated in the dark at 25 °C. However, previous incubationin the dark for 30 days at 4 °C partially substituted forthe light requirement. The possible mechanism of breakage ofseed dormancy in Talinumis discussed in relation to these andother findings. Talinum triangulare (Jacq.), Willd, light, photoperiod, seed germination     

Characteristics of Sugar, Amino Acid and Phosphate Release from the Seed Coat of Developing Seeds of Vicia faba and Pisum sativum

After removal of the embryo from developing seeds of Vicia fabaL. and Pisum sativum L., the ‘empty’ ovules werefilled with a standard solution (pH 5.5). Seed coat exudatesof both species were collected during relatively long experiments(up to about 12 h) and the concentration of sugar (mainly sucrose),amino acids and phosphate in the exudate measured. A discussionis presented on the amino acid/sugar ratio and the phosphate/sugarratio in the seed coat exudate. A pretreatment (15 min) withp-chloromercuribenzenesulphonic acid (PCMBS) reduced the releaseof sugar, amino acids and phosphate from broad bean seed coats.After excision of ‘empty’ ovules of Vicia faba andPisum sativum from the maternal plant, 2–4 h after thistreatment a strong difference became visible between sucroserelease from excised seed coats and sucrose release from attachedseed coats. Similarly, when the rate of phloem transport ofsucrose into an ‘empty’ ovule of Vicia faba or Pisumsativum was reduced by a sub-optimal mannitol concentrationin the solution, a reduced rate of sugar release from the seedcoat could be observed. Excision and treatment with a sub-optimalmannitol concentration reduced the release of amino acids toa lesser extent than for sucrose. These treatments did not reducethe rate of phosphate release from the seed coat. Key words: Seed development, Seed coat exudate, Phloem transport     

Phenotypic Mutations Induced During Storage in Barley and Pea Seeds

Previous work on chlorophyll-deficiency mutations in pea andbarley has shown that a significant increase in mutations isinduced by storing seeds under various conditions which leadto losses of viability to about 50 per cent. The work here showsthat a detectable increase in mutation frequency is also associatedwith much smaller losses of viability. Pea seeds were storedat 35 °C and 16.5 per cent moisture content for 40 and 57d when viability fell from 99 to 93 and 82 per cent, respectively.At the same time mutation frequency (percentage of seeds containingrecessive point mutations) increased from 1.62 per cent in thecontrol treatment to about 3 to 4 per cent. Barley seeds at15.5 per cent moisture content were stored at 50 °C for42 and 54 h, and at 35 °C for 28 and 39 d. During theseageing treatments viability fell from 98 to 75, 26, 93 and 48per cent respectively and the mutation frequency increased fromzero to between about 0.3 to 0.9 per cent. In both species theinduction of mutation by ageing treatments was significant butthe differences between the various ageing treatments were not.It is concluded that there is probably no safe threshold lossof viability which completely avoids mutation, and these resultssupport the view that for genetic conservation seeds shouldbe stored under conditions which minimise loss of viability. Pisum sativum L., pea, Hordeum distichum L., barley, mutation frequency, seed storage, seed viability