The Effects of Temperature and Moisture on the Induction of Genetic Changes in Seeds of Barley, Broad Beans, and Peas during Storage

Seeds were ‘aged’ by treatments with various combinationsof temperature and moisture content so that viability was reducedto about 50 per cent. It was shown that such treatments inducednon-heritable morphological abnormalities in about 10 per centof the bean plants and 5 per cent of the pea plants; but nonon-heritable morphological abnormalities were observed in thebarley plants. In all species there was an increased frequencyof pollen abortion in the plants produced from ‘aged’seeds. From the examination of second and third selfed generationsit was shown that single-recessive and double-recessive chlorophyll-deficiencymutations of various types were usually induced in sectors ofabout 1–3 per cent of the plants grown from treated seeds. The evidence from pollen-abortion supports previous cytologicalevidence that, under most storage conditions, for a given lossof viability, a predictable amount of chromosome damage is foundin the surviving seeds, irrespective of how fast viability waslost or which environmental factor was mainly instrumental inthe loss of viability. A comparison of the frequency of chlorophyllmutations produced under different storage treatments was notas quantitatively precise, but this evidence also is at leastcompatible with the thesis that loss of seed viability is associatedwith chromosome damage which is reflected in gene mutation.It is concluded, therefore, that percentage seed viability isa good index of the loss of genetic purity resulting from mutationswhich accumulate during storage.     

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     

Loss of Viability in Lettuce Seeds and the Accumulation of Chromosome Damage under Different Storage Conditions

Loss of seed viability in lettuce (Lactuca sativa L.) duringstorage is associated with an increase in the frequency of cellsin the surviving seeds showing chromosome damage during firstmitoses. The relation is linear when probit of the frequencyof aberrant cells is plotted as a function of probit percentagenormal germination. The slope of the relation, however, variesaccording to moisture content so that the proportion of aberrantcells for any given loss of germination increases with decreasein moisture content over the range 13.0–5.5 per cent.At 3.3 per cent moisture content, however, the proportion ofaberrations was no greater than at 5.5 per cent moisture content;and at 18.1 per cent moisture content the proportion was noless than at 13.0 per cent moisture content. Despite these differences,the increase in chromosomal aberrations per unit time for agiven temperature was always less the lower the moisture content.Diplontic selection markedly reduced the frequency of chromosomalaberrations and eliminated the differences in these frequenciesbetween the different storage treatments. But even after fiveweeks' growth, root tips from aged seed still contained abouttwice as many aberrant cells as compared with similar root tipsderived from the original seed stock. Studies on the frequencyof recessive mutations indicated that excessive amounts of heritablemutations were not present in the progenies of aged seed, evenwhen stored at moisture contents as low as 5.5 per cent. Allthis and other evidence reinforces the view that orthodox seedsfor genetic conservation should be stored at not more than about5 per cent moisture content, and that even lower moisture contentsare worth considering. The results also emphasise the need formaintaining a high regeneration standard, i.e. the percentageto which seed viability is allowed to fall during storage beforethe seed stock is regenerated. Lactuca sativa, lettuce, seed storage, seed viability, chromosomal aberrations, phenotypic mutations     

The Influence of Temperature, Moisture, and Oxygen on Period of Seed Viability in Barley, Broad Beans, and Peas

The viability of seeds of barley, broad beans, and peas hasbeen examined in hermetic storage over a range of temperaturesfrom 25? to 45? C and over a range of moisture contents fromabout 12 to 18 per cent. It was found that the survival curvesunder nearly all conditions can be described as negative cumulativenormal distributions. Under very severe storage conditions,however, when the mean viability period is of the order of aweek or less, the survival curves may become slightly skew.The spread of the distribution is linearly proportional to themean viability period. There is a negative linear relationshipbetween log mean viability period and both temperature and moisturecontent. Because of these relationships it is possible to predictthe percentage germination of these species after any givenperiod under a wide range of storage conditions. This patternof loss of viability in barley, broad beans, and peas is consistentwith that previously shown for wheat and rice. Oxygen has been shown to have a deleterious effect on the viabilityof barley, beans, and peas. Most of the deleterious effect isproduced by increasing the oxygen level from 0 per cent (nominal)to 21 per cent at atmospheric pressure; a further increase to100 per cent has comparatively little effect. The deleteriouseffect of oxygen is more pronounced at high moisture contents.Experiments at a low moisture content (12 per cent) have demonstratedthat the effect of oxygen is independent of the activity ofmicro-organisms. There is also some indication that the effectof oxygen is independent of the rate of seed respiration. The gas-exchange of pea seeds has been investigated in hermeticstorage at 25? C and 18.4 per cent moisture content. The seedsshowed a constant rate of gas-exchange and a constant R.Q. (0.63)over a period of time during which the concentration of oxygendecreased from 21 to I.4 per cent and the carbon dioxide concentrationrose from 0.03 to 12 per cent.     

Effects of Temperature, Moisture, and Oxygen on the Induction of Chromosome Damage in Seeds of Barley, Broad Beans, and Peas during Storage

The effects of temperature, seed moisture content, and oxygenlevel on the production of chromosome aberrations during seedstorage have been investigated. It has been found that an increasein any of these factors increases the rate of loss of seed viabilityand that any treatment which leads to a loss of viability alsoleads to an accumulation of aberrant cells in the embryo. Undermost storage conditions, irrespective of the combination offactors which leads to loss of viability or the rate at whichviability is lost, the relationship between percentage viabilityand mean frequency of aberrant cells in the surviving seed populationis always the same. Under very severe storage conditions, whichresult in half-viability periods of about a week or less, however,the relationship is altered so that for any given percentageviability the mean frequency of aberrant cells in the survivingseeds is less than is typical of more normal storage conditions.In all treatments (except the most severe) the curve showingmean frequency of aberrant cells in surviving seeds againsttime eventually became asymptotic to a critical value peculiarto the species. It is suggested that these results are compatible with the hypothesisthat under most storage conditions death of the embryo is theresult of the accumulation of nuclear damage which is reflectedin chromosome breakage; it is emphasized, however, that thevisible chromosome breakage itself is probably not criticalto the seed's survival. When the nuclear damage has reacheda critical level, of which the frequency of aberrant cells isan index, the seed is no longer capable of germinating. Undervery severe storage conditions, additional factors appear tobe involved. An effort was made to test the frequently suggested hypothesisthat chromosome aberrations produced during the ageing of seedsare the result of the accumulation of automutagenic substances.Attempts to demonstrate the presence of mutagenic substancesin water-, ethanol-, and ether-extracts of fresh and aged seedsgave negative results. Aqueous extracts showed antimitotic andgermination inhibitory activity, but this was the same fromboth aged and fresh seeds.     

Logarithmic Relationship between Moisture Content and Longevity in Sesame Seeds

The relationship between seed moisture content and seed longevityin sesame (Sesamum indicum L.) in hermetic storage at 50 °Cis logarithmic. The logarithmic relationship is maintained from15 per cent down to 2 per cent – the lowest moisture contenttested — but above 15 per cent this ‘air-dry’relationship no longer holds since further increase in seedmoisture content does not reduce longevity. Tentative estimatesof constant values for the improved seed viability equationare provided, and implications for long-term storage are discussed. Sesame, Sesamum indicum L., seed storage, improved viability equation, seed moisture content, seed longevity prediction     

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     

Improved Equations for the Prediction of Seed Longevity

Equations for predicting seed longevity in storage have beenimproved so that they now take into account variations withina species in initial seed quality—which is affected bygenotype and pre-storage environment—and so that theyare more accurate over a wider range of storage environmentsThese improvements have been incorporated into a seed viabilitynomograph for barley (Hordeum distichum L.) which may be usedto predict percentage viabihty of any seed lot after any timein any storage environment within the range –20 to 90°C and 5–25 per cent moisture content. Applicationsof the improved equations to seed drying and to long-term seedstorage for genetic conservation are discussed. Hordeum distichum L., barley, seed viability, seed longevity prediction, seed storage, seed drying, storage temperature, seed moisture content, genetic resources conservation     

Nucleotide Levels and Loss of Vigour and Viability in Germinating Wheat Embryos

At an optimal germination temperature loss of viability in wheatseed is reflected in reduced levels of nucleoside triphosphatesand nucleotide sugars in the embryo compared to the levels foundin high viability embryos during the early hours of imbibition.These differences are magnified on germination at 10° Cand are accompanied by a greatly reduced rate of protein synthesisin embryos of low viability compared with the correspondingrate in embryos of high viability. Loss of vigour precedes lossof viability in seed but differences in biochemical parametersbetween high and medium vigour seed do not become apparent unlessgermination under stress conditions, e. g. low temperature,occurs. Under these stress conditions, wheat seed of reducedvigour can be distinguished from high vigour seed lots of similarviability by the decreased levels of nucleotides and nucleotidesugars found in embryos from reduced vigour seed lots duringthe early hours of imbibition and by reduced rates of proteinsynthesis in these embryos during this same period. Nucleotideanalysis is a potentially useful method for rapid assessmentof the viability or vigour of a seed lot. Key words: Nucleotides, Vigour, Viability, Wheat     

The effect of iprodione on seed germination and seedling emergence in onion

Iprodione seed treatment at 100 g a.i./kg seed had little effect on final emergence in peat-based compost at 60% moisture content (wet weight basis) but caused up to 1.5 days' delay in the time to 50% of the final emergence (t₅₀). The effect was independent of initial seed viability, the conditions and duration of seed storage after treatment and germination temperature. Raising the compost moisture content to 70% resulted in a 9% reduction in final emergence and a 3 day increase in t₅₀. Iprodione had a greater effect in a filter paper test, reducing final emergence by up to 24% and increasing t₅₀; the effects were again independent of initial seed viability and storage conditions.     

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     

The Influence of Temperature and Moisture on Seed Viability Period in Barley (Hordeum distichum L.)

Storage experiments were carried out on barley seed (Hordeumdistichum L.) lasting from 1 min to 926 days, including 52 hermeticstorage environments covering a range of temperatures from 3to 90 °C and 5·5 per cent to 24·6 per centmoisture content (f. wt basis). Over the entire range of conditionssurvival curves conformed to negative cumulative normal distributionsand, for any given measure of longevity, e.g. half-viabilityperiod, longevity was roughly proportional to the negative exponentof both temperature and moisture content. Although previouslyreported viability equations were adequate to describe theserelationships over restricted ranges of environments, over theextended range of conditions tested here it was shown that therelationship between log seed viability and temperature is infact slightly convex, whilst that between log seed viabilityand moisture content is slightly concave. An improved viabilityequation was applied which takes into account those curvaturesand, at the same time takes into account the initial viabilityof a seed lot which reflects pre-storage deterioration. Thefit was excellent and thus it is now possible to predict percentageviability of any lot of barley seed after any storage periodunder a very wide range of conditions. Hordeum distichum L., barley viability, seed longevity, seed storage, moisture content of barley seed, temperature, influence of seed viability     

Seed Viability Constants for Lettuce

Seeds of two lettuce cultivars (Lactuca sativa L., cv. Meikoninginand cv. Grand Rapids) were hermetically stored with constantmoisture contents ranging between 3.6 and 17.9 per cent (freshweight basis) at constant temperatures ranging between 5 and75 °C. The decline with time in percentage germination andpercentage normal seedlings was determined for each storagetreatment. The data were fitted to an equation which containsthe constants: K₁, the probit of the initial percentage germinationor normal seedlings; K_E, a species constant; C_W, the constantof a logarithmic moisture term; C_H, the constant of a lineartemperature term and C_Q, the constant of a quadratic temperatureterm. Regression analysis of data from storage periods up to5.5 years at temperatures of 5–75 °C and seed moisturecontents of 3.6–13.6 per cent yielded the following values:K_E= 8.218, C_W=4.797±0.163, C_H=0.0489±0.0050 andC_Q=0.000365±0.000056. Although this equation consistentlyprovided a better fit, simplified equations, assuming eithera log-linear relationship between seed longevity and temperature,or a log-linear relationship between seed longevity and bothmoisture content and temperature, accounted for more than 94per cent of the variation at the restricted temperature rangeof 5–40 °C. Longevity of the same seed lots at sub-zero temperatures (–5,–10 and –20 °C) was studied in separate tests.Freezing damage, resulting in abnormal seedlings in the germinationtest, occurred at –20 °C when the moisture contentof the seeds exceeded 12 per cent. No decline in percentagenormal seedlings was observed after a storage period of 18 monthsor longer at –20 °C, provided the seed moisture contentdid not exceed 9.5 per cent. For seeds stored at –5 and–10 °C with 9.6–12.5 per cent moisture content,the observed rate of decline of percentage normal seedlingswas adequately predicted by the viability equation, using theabove values for the constants. This suggests that for low moisturecontents the viability equation can be applied to estimate longevityat sub-zero temperatures. Lettuce, Lactuca sativa (L.), seed longevity, seed storage, viability constants, storage conditions     

Biochemical changes induced in seeds of <Emphasis Type="Italic">Brassicaceae</Emphasis> wild species during ageing

The aim of the present study was to determine whether the loss of seed germination capacity and vigour in seeds of four wild Brassicaceae species (Brassica repanda, Moricandia arvensis, Rorippa nasturtium-aquaticum and Sinapis alba) during ageing at 45°C and 90% relative humidity was related to changes in lipid peroxidation and membrane integrity. For all of the species, ageing reduced the final germination percentage and increased the length of time required to reach 50% of final germination (T ₅₀). Large differences in longevity were observed among the species. The times required for viability to be reduced to 80 and 50% of maximum germination (P80 and P50) were the lowest for B. repanda, and these values were two times longer for M. arvensis and R. nasturtium-aquaticum and five times longer for S. alba compared with B. repanda. A loss of seed viability was not associated with malondialdehyde accumulation, suggesting that lipid peroxidation did not cause seed deterioration under these conditions. However, the conductivity test effectively detected seed deterioration in these wild Brassicaceae species, and membrane permeability correlated with both germination and vigour loss. This correlation may provide a valuable mean for early detection of seed viability in wild Brassicaceae species.     

Freezing injury to ovules, pollen and seeds in winter rape

Winter rapeseed (Brassica napus, cv. Samouraï) flowersearly in spring and, under field conditions, short freezingperiods can occur. Unacclimatized plants were freeze-stressed(–3°C for 4 h) at different developmental stages ofbuds, open flowers and seeds. The dissection of pistils from stressed plants showed that freezingresults in shrivelled ovules. We assessed freezing injury onthe basis of per cent of shrivelled ovules: ovule sensitivitybegins early (8 d before anthesis) but increases up to anthesis.Crosspollination of stressed pistils with non-stressed pollenshowed that recording of freeze-injured ovules is a good methodfor early estimation of the effect of stress on seed yields. On the other hand, stress does not reduce the viability of pollen,except when it was applied at the binucleate pollen stage. Useof frozen pollen x nonstressed pistils has little effect onseed yields. Freeze injury on seeds was assessed by seed filling:seeds are very susceptible just after fertilization until 20d after fertilization (DAF). Freezing seems to inhibit seedfilling. A germination test of stressed seeds during their developmentindicated that embryo viability is not affected if the stressoccurs after 35 DAF. As the embryos develop, resistance to stressincreases. Key words: Brassica napus, rapeseed, freeze injury, pollen and ovule, seed filling     

Water and Seed Survival

Between about –350 and –14 MPa the rate of lossof viability in orthodox seeds is a positive function of waterpotential. The relative effect of water potential has been analysedin an oily seed (lettuce) and a non-oily seed (barley) and foundto be more or less identical. The lower limit for the relationin various species coincides with a seed moisture content (wetbasis) between about 2 and 6%. Below this level there is littleor no improvement in longevity with reduction in moisture content.The upper limit coincides with moisture contents of between15 and 28%, depending on whether the seeds are oily or non-oily.A water potential of about –14 MPa is the threshold forrespiration which increases more-or-less linearly with increasein water potential above this level. Above this threshold, andproviding oxygen is available to sustain respiration, seed longevityincreases with increase in water potential except that, unlessthe seeds are dormant, germination may be initiated at a waterpotential of about –1·5 to –0·5 MPa.In the absence of oxygen there may be a slight further declinein longevity with increase in water potential above –14MPa before longevity reaches a minimum value Since they cannot be dried very much without immediate lossof viability, recalcitrant seeds survive longest in the presenceof oxygen at maximum water potential commensurate with preventinggermination. The threshold water potential for immediate lossof viability has not been determined for most species but itis probable that it is close to the water potential typicalof the permanent wilting point in these plants, say –2MPa Lactuca saliva L., lettuce, Hordeum oulgare L., barley, seed storage, moisture content, relative humidity, water potential, temperature, oxygen     

The Influence of Genotype, Temperature and Moisture on Seed Longevity in Chickpea, Cowpea and Soya bean

Seed of three chickpea (Cicer arietinum L.), three cowpea [Vignaunguiculata (L.) Walp.] and four soya bean [Glycine max (L.)Merr.] cultivars were hermetically stored for up to 2 yearsin various constant environments which included temperaturesfrom —20 to 70 °C and moisture contents (fresh weightbasis) from 5 to 25 per cent. In all cases the survival curvescould be described by negative cumulative normal distributions.The longevity of the various seed lots differed but the valueof the standard deviation (the reciprocal of which gives theslope of the survival curve when percentage germination is transformedto probit) was the same for all cultivars within a species whenstored under similar conditions. Within each species the relativeeffects of moisture and temperature on longevity did not differsignificantly between cultivars. In all three species therewas a negative logarithmic relationship between seed moisturecontent and longevity, but the relative effect of moisture contentdiffered between the species: differences in the longevity ofsoya bean seed as a function of moisture content were less thanfor either cowpea or chickpea. The relative effect of temperatureon seed longevity did not differ between the three species,and the seed of all three species showed increasing temperaturecoefficients for the change in rate of loss of viability withincrease in temperature. The complete pattern of loss in viabilityin all three species can be described by a single equation whichwas developed for barley and has also been shown to apply toonion seed. The constants applicable to the three grain legumeshave been calculated so that it is now possible to predict percentageviability of any seed lot of these species after any storageperiod under a very wide range of storage conditions. Cicer arietinum L., chickpea, Glycine max (L.) Merr., soya bean, Vigna unguiculata (L.) Walp., cowpea, seed longevity, seed storage, moisture content, temperature     

Presevvation of Xanthomonas campestris in Brassica oleracea seeds

Summary Brassica oleracea seeds were sterilized by gamma radiation and sodium hypochlorite washing. Xanthomonas campestris was inoculated into the seeds by incubating, under vacuum, a suspension of the bacteria with the seeds. After thorough washings with sterile distilled water, the seeds retained about 13 000 cells per seed. The seeds were maintained at 4°C during 21 months, during which the viability of the bacteria and their capacity to produce xantham gum in shake flasks, were evaluated. Bacterial viability showed oscillations but after 20 months it was 10% of the initial. When these seeds were used as a pre-inoculum for a culture to produce xanthan, the final polymer concentration increased slightly with time of seed storage and the final broth viscosity was fairly constant. The specific polymer production (per weight of final bacterial cells) increased about three-fold after 21 months of experimentation. The method, besides being able to produce xantham in quantity and quality, has the advantages of an easy inoculation procedure, no need for transfers, less contamination risk and improved growth rate of the bacteria in the inoculation medium. Correspondence to: E. Galindo     

The Influence of Gaseous Environment on the Storage Life of Araucaria hunsteinii Seed

The effect of various gas mixtures on the longevity of hydratedseeds of Araucaria hunsteinii K. Schum. was assessed under controlledconditions. The length of storage life decreased as oxygen concentrationwas reduced from 21 to zero per cent. No effect of carbon dioxideon seed longevity was detected within the range 1–50 percent when combined with 10 or 21 per cent oxygen. Ethylene at0.01 per cent, and sealed foil or polyethylene bag storage reducedthe period of seed germinability compared with that for 21 percent oxygen. Ethanol accumulation took place in stored seedswhen the environmental oxygen concentration was below a thresholdvalue which lay between 1 and 5 per cent. It is proposed that the observed effects of gases on longevityof hydrated seeds may be mediated through an influence on aerobicrespiration rate. Practical implications of the results areconsidered. Araucaria hunsteinii, Klinkii pine, seed longevity, seed storage, gas environments, oxygen, carbon dioxide, ethanol accumulation