The Effects of Priming and Ageing on Resistance to Deterioration of Tomato Seeds

The influence of seed priming and ageing treatments on viabilityand rate of germination of tomato (Lycopersicon esculentum Mill.)seeds was examined under both long-term and controlled-deteriorationstorage conditions. Seeds of a single lot of tomato were eitherprimed or aged to increase or decrease the rate of germination(Argerich and Bradford, 1989). They were then stored at 6% moisturecontent (dry weight basis) at either 4 ?C or 30 ?C for 1 year.Both viability and germination rate were unaffected by eitherstorage temperature in control seeds, or by 4 ?C storage inprimed or aged seeds. At 30 ?C, however, viability and germinationrate of primed and aged seeds was markedly reduced after 6 monthsof storage. The temperature dependence of the germination rateand the spread of germination times within the population wasalso adversely affected by high temperature storage, particularlyfor primed seeds. Under controlled deterioration conditions(13.5% moisture content and 50 ?C), the rate of loss of viabilitywas greater for primed seed than for control or aged seeds.The relationship between seed viability and the mean germinationrate, however, was not influenced by the seed treatments. Thesedata are analysed in relation to current models of seed deteriorationduring storage and seed repair during priming. The results indicatethat enhancement of seed germination rates by priming treatmentssimultaneously lowers the resistance of seeds to deterioration.Primed tomato seeds must, therefore, be considered to be vigorousseeds with a reduced storage life. Key words: Tomato, controlled deterioration, seed germination rate, seed viability     

The Effects of Priming and Ageing on Seed Vigour in Tomato

A comparison was made of the effects of seed priming or ageingtreatments on the performance of tomato (Lycopersicon esculentumMill. cv. UC204C) seeds according to a number of indices ofseed vigour. A single lot of tomato seeds was primed in 120mol m^–3 K2HPO4 + 150 mol m^–3 KNO₃ for 5 d at 20?C, or aged at 13% moisture content (dry weight basis) and 50?C for 6 d. Germination percentage (>98%) was unaffectedby priming and reduced to 85% by ageing. X-ray photographs andlongitudinal sections revealed the formation of free space surroundingthe embryo in dry primed seeds, which was not evident in controlor aged seeds. Priming increased the rate of germination atall temperatures above the base temperature (T_b), while ageingdecreased it. T_b was unaffected by priming and only slightlyincreased by ageing. The variation in individual times to germinationwas approximately doubled in both primed and aged seed comparedto the control, based upon the slopes of probit germinationpercentage versus log thermal time curves. Root growth aftergermination tests and seedling growth in both greenhouse andfield tests were not influenced by either priming or ageing.The conductivity test was found to be unreliable as a vigourtest for tomato seeds. The results identify several indiceswhich can be used to quantify seed vigour in tomato. They alsoillustrate that seed priming can enhance seed performance accordingto some criteria, while having no effect or decreasing qualityaccording to other criteria. Seed vigour can apparently be separatedinto various components which can be independently influencedby seed enhancement treatments. Key words: Tomato, seed germination rate, seed priming, seed vigour     

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     

Evaluation of Seed Storage Conditions and Genetic Diversity of Four Crucifers Endemic to Spain

Traditional assays (germination and vigour), as well as biochemicalassays using isozyme loci, were performed to evaluate the effectof different seed storage conditions in Coincya rupestris, Iberispectinata, Moricandia moricandiodes and Vella pseudocytisus.Seeds had been stored for 24–30 years under long-term(LT) and short-term (ST) conditions. Fresh seeds of the samespecies from natural populations (NP) were also collected in1996 and used for comparison. Isozyme loci were used as markersof genetic deterioration and to determine the genetic diversityof samples. The germinability, vigour and enzyme activity ofLT samples were comparable to values obtained using freshlycollected NP samples, showing the effectiveness of the long-termstorage conditions. Copyright 1999 Annals of Botany Company Endemics, genetic diversity, germination, isozymes, seed ageing, seed storage, vigour.     

Catalase is a key enzyme in seed recovery from ageing during priming

Ageing induces seed deterioration expressed as the loss of seed vigour and/or viability. Priming treatment, which consists in soaking of seeds in a solution of low water potential, has been shown to reinvigorate aged seeds. We investigate the importance of catalase in oxidation protection during accelerated ageing and repair during subsequent priming treatment of sunflower (Helianthus annuus L.) seeds. Seeds equilibrated to 0.29 g H₂O g⁻¹ dry matter (DM) were aged at 35 °C for different durations and then primed by incubation for 7 days at 15 °C in a solution of polyethylene glycol 8000 at −2 MPa. Accelerated ageing affected seed germination and priming treatment reversed partially the ageing effect. The inhibition of catalase by the addition of aminotriazol during priming treatment reduced seed repair indicating that catalase plays a key role in protection and repair systems during ageing. Ageing was associated with H₂O₂ accumulation as showed by biochemical quantification and CeCl₃ staining. Catalase was reduced at the level of gene expression, protein content and affinity. Interestingly, priming induced catalase synthesis by activating expression and translation of the enzyme. Immunocytolocalization of catalase showed that the enzyme co-localized with H₂O₂ in the cytosol. These results clearly indicate that priming induce the synthesis of catalase which is involved in seed recovery during priming.     

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     

Prolonged aerated hydration for improvement of seed quality in Brassica oleracea L.

Seeds of cauliflower cv. Hipop and Brussels sprouts cv. Asmer Aries were aged at 20% moisture content for 24 h; all seeds retained a germination of over 70% after ageing although the mean germination time increased. Prolonged aerated hydration for up to 32 h at 20°C followed drying resulted in improved performance of both unaged and aged cauliflower seeds and aged Brussels sprouts. Thus, all seed showed reductions in the mean germination time to the extent that after 32 h hydration the aged cauliflower seeds performed as well as high quality unaged seed. The improvement of aged seeds was also revealed an increase in germination after the controlled deterioration test following up to 24 h (cauliflower) or 32 h (Brussels sprouts) aerated hydration. This increase was indicative of a decrease in the extent of deterioration present after aerated hydration. Deleterious effects of prolonged hydration were observed in Brussels sprouts after 32 h although these may be explained desiccation injury after treatment since radicle emergence had occurred during hydration. The improvements in seed performance may be explained the activation of metabolic repair occurring during the early part of the hydration period therereducing the extent of deterioration that has been sustained during ageing, with further improvements due to the advancement of the germination process.     

The Role of Anaerobic Respiration in Germinating Muskmelon Seeds: I. IN RELATION TO SEED LOT QUALITY

Five muskmelon (Cucumis melo L.) cultivar seedlots from a commercialsource and freshly produced seeds of two cultivars, when artificiallyaged, were found to differ in their viability and vigour asdetermined by germination tests. Furthermore, the various commercialseedlots without ageing also exhibit a range of deteriorationlevels. Low vigour seeds had higher respiratory quotient valuesthan the high vigour seeds as a result of a higher level ofCO₂ production. This high level of CO₂ evolution in low vigourseeds may have been due to anaerobic respiration. Levels of acetaldehyde and ethanol produced by imbibing seedswere negatively associated with seed viability and vigour. After6 h of imbibition low vigour seeds produced significantly moreethanol and acetaldehyde than high vigour seeds. After 24 hof imbibition, ethanol continued to accumulate in the commercialseedlots up to 10-fold the amount produced after 6 h of imbibition,whereas, acetaldehyde levels increased less. However, in thefreshly produced, artificially aged seeds (except the most extremeageing), ethanol levels were reduced and no acetaldehyde productioncould be detected, indicating re-utilization of ethanol. It is suggested that ethanol production in the first hours ofimbibition can be used as a reliable index to predict germinationin muskmelon seedlots. Key words: Germination, Cucumis melo L., Seeds, Anaerobic respiration     

The influence of aerated hydration seed treatment on seed longevity as assessed by the viability equations

Aerated hydration (AH) treatments of cauliflower seeds for 12 h (12AH) and 28 h (28AH) at 20 degrees C resulted in improved or reduced storage potential of low or high vigour seeds, respectively. Seeds were stored at their initial seed moisture content (mean 5.5% mc) or at 12% mc at 10 degrees C for 12 months and at 20 degrees C for 4 months. The improved longevity of low vigour seeds was associated with increased K(i) (initial seed viability) and a reduced rate of deterioration (1/sigma) whereas the K(i) of high vigour seeds fell after 28AH and the rate of deterioration increased such that the time to lose one probit of viability decreased from 28.7 to 5.3 months at 10 degrees C and from 10.4 to 1.2 months at 20 degrees C. The improved K(i) of low vigour seeds could be explained by the reduction in the extent of deterioration after AH, as indicated by the increase in germination after cotrolled deterioration (CD), and the possible activation of metabolic repair during treatment. In contrast the reduced germination after CD of AH-treated high vigour seeds was indicative of deterioration as a result of treatment. Both high and low vigour seeds contained constitutive levels of ss-tubulin which increased during AH treatment, the increase being greater in high vigour seeds. High vigour seeds also showed an increase in the proportion of nuclear DNA present as 4C DNA, from 3% (untreated seeds) to 26% (28AH), indicative of germination advancement from the G(1) to G(2) phase of the cell cycle during treatment. This higher proportion of 4C DNA is correlated with the increased sensitivity of seeds to drying and/or storage after AH, leading to their reduced K(i) and storage potential. In contrast, there was little change in %4C in low vigour seeds. Priming in polyethylene glycol (PEG, -1.0 MPa) for 5 d or 13 d also improved the longevity of low vigour seeds stored at their initial and 12% mc at 10 degrees C for 8 months, as reflected in their laboratory and CD germination. In this case, however, the improved longevity of the low vigour seeds following 13 d priming was associated with an increase in 4C DNA from 4% (dry control) to 56% after treatment. The germination of both untreated and primed high vigour seeds remained high throughout the storage period. Increases in the rate of germination (decreased mean germination time) observed after all AH and PEG treatments were not consistently associated with an increase in the proportion of nuclei containing 4C DNA.     

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     

Priming and accelerated ageing affect L-isoaspartyl methyltransferase activity in tomato (Lycopersicon esculentum Mill.) seed

Damage and degradation of cellular proteins is observed duringage-induced seed deterioration. L-Isoaspartyl protein methyltransferase(EC 2.1.1.77 [EC] ) is an enzyme hypothesized to play a role in limitingand repairing age-induced damage to proteins. Tomato (Lycopersiconesculentum Mill. ‘New Yorker’) seeds were assayedfor changes in L-isoaspartyl methyl-transferase activity duringaccelerated ageing and after osmotic priming. Accelerated ageingof seeds for 1–4 d at 45C and 100% relative humidityreduced germination from 94% to 71%, increased the mean timeof germination (MTG) from 2.4 to 5.8 d, and was accompaniedby a correlative decrease in L-isoaspartyl methyltransferaseactivity (r²=0.90). Aged and untreated seeds were primed for7 d at 20C in darkness using aerated solutions of 3% KNO₃ orpolyethylene glycol 8000 (PEG) with equivalent osmotic potential(–1.25 MPa). Priming with KNO₃ decreased the MTG, butdid not improve germination percentage for untreated seeds.Priming did not affect L-isoaspartyl methyltransferase activityin untreated seeds, but restored activity in aged seeds primedin KNO₃ to levels near that of untreated seeds. Priming withPEG did not effectively improve the MTG or increase L-isoaspartylmethyltransferase activity. During germination, L-isoaspartylmethyltransferase activity remained constant for 48 h post-imbibitionand then declined, suggesting that the enzyme was developmentallyregulated and inactivated or degraded as radicle emergence occurred. Key words: L-Isoaspartyl methyltransferase, protein repair, seed priming, accelerated ageing, Lycopersicon esculentum     

Changes in Lipids of Bean Seeds (Phaseolus vulgaris) and Corn Caryopses (Zea mays) Aged in Contrasting Environments

Seeds of French bean (Phaseolus vulgaris L. cv. The Prince)and caryopses of sweet corn (maize; Zea mays L.F1 hybrid Firstof All) were stored in environments of 79% relative humidityand 25 °C, 80% r.h. and 40 °C or 100% r.h. and 45 °C,giving ageing (loss of gcrminability and vigour) over periodsof months, weeks or days, respectively. The relationship betweenchanges in lipids and changes in germinability or vigour wasunaffected in general by the speed of ageing. In the corn caryopsesthere was no evidence of hydrolysis of phospholipids or peroxidationof fatty acids during ageing. In the bean seeds phosphatidicacid increased during the ageing period and phosphatyl cholinedeclined. The percentage of fatty acids as hnolenic acid initiallyfell during bean ageing, but in the slower ageing conditionsit rose again as germination reached zero. In bean, the presenceof phosphatidic acid could be a sensitive indictator of lossof vigour, but relative proportions of the different fatty acidswould be a misleading indicator of quality. Rapid artificialageing may be an adequate model, in some species, of ageingat moderate speeds and of ageing under some ambient conditions. French bean (Phaseolus vulgaris L. cv. The Prince), sweet corn (maize, Zea mays L., Fl hybrid First of All), seed ageing, phospholipids, fatty acids     

Responses of Vegetable Seeds to Controlled Hydration

Leek, onion and carrot seeds were imbibed in water and in solutionsof polyethylene glycol (PEG) 6000 over the range –0.5to –4.0 MPa osmotic potential, for periods up to 28 dat 15 C. Seeds started to germinate after 7 and 14 d at –0.5MPa and –1.0 MPa PEG, respectively, but in the lattercase, germination did not exceed 5%. No germination occurredin solutions of lower (more negative) osmotic potential. Seedmoisture content increased with osmotic potential in all threespecies and the relationships were unaffected by the durationof imbibition in solutions of –1.0 to –4.0 MPa,though leek seeds had higher moisture contents than the otherspecies for any given osmotic potential. Linear relationships between response to priming (differencebetween mean germination times of primed and untreated seeds)and seed moisture content were obtained for each species, positiveresponses being obtained above 30–35% seed moisture contentwith optima at 46, 44.5 and 44% seed moisture contents in leek,onion and carrot, respectively. Priming had no effect on embryovolume or cell number per embryo in leek and onion. Carrot embryovolume increased by 43% and cell number per embryo doubled inprimed compared with untreated seeds, whereas seeds imbibedin water showed only a slight increase in cell number per embryoat the stage when radicles were beginning to penetrate the seedcoat. Allium cepa L. cv. Rijnsburger Robusta, onion, Allium porrum L. cv. Winterreuzen, leek, Daucus carota L. cv. Nantaise, carrot, germination, priming, polyethylene glycol, seed moisture, cell number, embryo volume     

Ageing and Osmotic Priming in Wheat Seeds: Effects Upon Certain Components of Seed Quality

Deteriorated wheat seed lots were osmotically primed and surface-dried,dried-back, then aged for two more weeks Osmotic priming didnot affect the final germination in any of the aged seed lots,but mean germination time was decreased The response of unagedseed lot to treatments was studied in the following the timecourses of germination, as well as L-[4, 5-²H]leucine and [6-³H]thymidineincorporation (as a measure of protein and DNA synthesis, respectively)into dissected embryos during the early hours of germinationIt was found that the variation in mean germination time andthe rate of synthetic reactions were related The greatest improvementin the components of seed lot quality was achieved after primingand surface drying The beneficial effects of priming were apparenteven after dehydration followed by two further weeks of ageingCumulative correlations between mean germination time and variousbiochemical tests in variously treated aged seed lots were highlysignificant The physiological processes involved in deteriorationand recovery and the potential application of biochemical testsin detecting vigour changes are discussed Triticum durum, seed, ageing, osmotic priming, mean germination time, protein DNA synthesis     

{beta}-Amylase Activity as an Index for Germination Potential in Rice

Seeds of different vigour also differ in their germination ability.In rice (Oryza sativa), this difference was correlated withthe level of incorporation of ³⁵S-methionine into 25-60% ammoniumsulphate precipitable material that was rich in amylase proteins.This protein fraction, from dry seeds, contained no -amylaseactivity. In contrast, ß-amylase activity was presentin all seed stocks capable of 99% germination, although thelevel was lower in seeds that grew slowly when germinated. Inlow viability low vigour stock (i.e. extensively deterioratedseeds) ß-amylase activity was absent. Alpha-amylaseactivity in all stocks was detected only after 24 h from thestart of imbibition. These results indicate that ß-amylaseactivity is reliable indicator of the germination ability ofrice seed stocks and of their vigour during germination.Copyright1995, 1999 Academic Press Rice (Oryza sativa L.,), germination, ß-amylase, -amylase, seed vigour     

Survival and Vigour of Lettuce (Lactuca sativa L.) and Sunflower (Helianthus annuus L.) Seeds Stored at Low and Very-low Moisture Contents

Seeds of lettuce (Lactuca sativa L.) and sunflower (Helianthusannuus L.) were stored hermetically at 35 °C with 11 differentmoisture contents between 1·3 and 6·9%, and between1·3 and 7·1% of fresh mass, respectively. Germinationand vigour (mean germination time, root length, seedling dryweight) were determined after storage for 0, 8, or 16 weeks(sunflower) or 0, 8, 16, or 48 weeks (lettuce) in these environmentsfollowed by various humidification treatments (to avoid imbibitioninjury). The range of seed storage moisture contents over whichdeterioration was minimized depended upon the criterion of deteriorationused, and varied somewhat between species. Comparison of theseranges for seeds stored for the longest durations showed thatfor some criteria seed performance was poorer (P < 0·05)at both the lowest and highest moisture contents investigatedthan at certain of the intermediate storage moisture contents(e.g, most rapid germination occurred in sunflower followingstorage at 2·2-4·7% moisture content), whereasfor other criteria all the drier storage moisture contents weresuperior to the more moist (e,g. greatest seedling growth occurredin sunflower following storage at 1·3-5·1% moisturecontent). But none of these results suggested that lettuce andsunflower seeds stored hermetically at 2·5-3·0%or 2·2-2·5% moisture content, respectively, wereless vigorous than at any other moisture content tested. Inboth species, these storage moisture contents are in equilibriumwith about 8-10% relative humidity (r.h.) at 20 °C, whichis similar to and indeed marginally less than the 10-13% r.h.recommended following earlier studies on the longevity of seedsin hermetic storage at much warmer temperatures. Thus, theseresults show no evidence that the optimum seed moisture contentfor storage increases with decrease in temperature, at leastover the range 35-65 °C, as has been suggested elsewhere.We conclude that the international recommendation for the long-termseed storage for genetic conservation at 5 ± 1% moisturecontent should not be revised upwardly, and that in situationswhere refrigeration cannot be provided storage at even lowermoisture contents is worthy of further investigation for thoseseeds in which desiccation at 20 °C to equilibrium at 10%r.h. results in moisture contents well below 5%.Copyright 1995,1999 Academic Press Helianthus annuus L., sunflower, Lactuca sativa L., lettuce, desiccation, seed storage, seed vigour     

Cell cycle inhibitors improve seed storability after priming treatments

Seed priming is a treatment that controls seed water content to partially activate germination processes such as metabolism but prevents full germination of the seeds. The treatment is well known to enhance seed performance, including germination, but sometimes reduces seed storability or longevity as a side effect. Toward developing a novel priming technique that can maintain seed longevity for a longer time period, chemicals that suppress the seed deterioration under a controlled condition were screened from 80 known biologically active compounds contained in the RIKEN NPDepo authentic library using Arabidopsis thaliana seeds. Seeds primed with mimosine, a cell cycle inhibitor, retained higher survival rate after a controlled deterioration treatment compared to seeds primed without the chemical. In addition, other cell cycle inhibitors such as aphidicolin, hydroxyurea and oryzalin had similar effects on the seed storability after priming. Our results suggest that progression of the cell cycle during priming is an important checkpoint that determines the storability of seeds after the treatment.

     

Investigation of the relationship between seed leachate conductivity and the germination of Brassica seed

Large differences in the leachate conductivities of single seeds after 24 h soaking were observed in five Brussels sprout cultivars. Within each cultivar non-germinable seeds tended to have higher conductivities than germinable seeds, but the conductivity above which seeds failed to germinate varied between cultivars. No conductivity reading or partition value could therefore be considered as an absolute boundary between germinable and non-germinable seeds to allow the prediction of seed germination for all seed lots of the same species. However, the mean conductivity for each sample was significantly correlated with germination (r = 0.83, P < 0.05) and was clearly higher for samples with lower germinations. Differences in the partition values for the five cultivars may have been attributable to the natural age of the seeds. Seeds of Asmer Aries artificially aged rapidly at 20% moisture content for up to 48 h at 45° C showed increasing leachate conductivity with increased time of ageing (r = 0.97, P < 0.01) and decreased germination (r = - 0.88, P < 0.05). Again however the values for germinable and non-germinable seeds of different artificial ages varied. Leakage from seeds was limited by the testa in the first 6 h of imbibition. Solute leakage from embryos of the five cultivars was therefore greater than those for intact seeds but a clear separation of germinable and non-germinable seeds was still not possible from the conductivity readings. The implications of the observed increases in conductivity with increased seed age and decreased germination for the development of a rapid vigour test for small seeded vegetables in conjunction with controlled deterioration are discussed.