Enzymatic Conversion of Volatile Metabolites in Dry Seeds during Storage

Dry seeds can transform volatile metabolites via enzymes pre-existingin them. The interconversion between acetaldehyde and ethanoloccurred when they were applied gaseously to seeds of rice,lettuce, pea, cocklebur during storage at different relativehumidities (RH) at either 23°C or –3.5°C. Interconversionsbetween the compounds decreased with decreasing RH from 75%to 12% regardless of seed species and storage temperature, butit was still detectable even at 12% RH at –3.5°C.However, the conversion from acetaldehyde to ethanol did notoccur when seeds were killed by heating prior to storage. 2-Methoxyethanol,a competitive inhibitor of alcohol dehydrogenase (ADH) (EC 1.1.1.1 [EC] ),suppressed the transformation from ethanol to acetaldehyde inlettuce seeds. Therefore, ADH pre-existing in dry seeds maybe involved in the interconversion between ethanol and acetaldehydein dry seeds. Propanal, also a substrate of ADH, could be transformedto propanol during storage. However, methanol, which is nota substrate of this enzyme, was hardly converted to formaldehyde.Ethylacetate, applied to seeds during storage, was hydrolyzedby both lettuce and rice seeds, and the amount of hydrolysisincreased with increasing RH. Similarly, other carboxylic esters,such as methylacetate, ethylformate and ethylpropionate, couldbe hydrolyzed by rice, pea, and lettuce seeds. Little hydrolysisof ethylacetate occurred in the seeds killed by heating beforestorage, suggesting that ethylacetate may be hydrolyzed by carboxylic-esterhydrolase (EC 3.1.1.1 [EC] ). On the other hand, the production ofethylacetate through esterification was demonstrated by additionof gaseous acetate and ethanol in lettuce seeds, which occurredmore greatly at 12% RH than at 75% RH. These findings were discussedin relation to the mechanism of seed aging. (Received September 3, 1994; Accepted November 14, 1994)     

Germination and Food Reserves in Bambarra Groundnut Seeds (Voandzeia subterranea Thouars) after Different Periods of Storage

Studies were made on bambarranut seeds (Voandzeia subterraneaThouars) after 0, 6, 12, 18 and 24 months of storage in gunnybags under laboratory conditions (25–35 °C). Seeddeterioration during storage was indicated by delayed germination,reduced germinability, reduced growth of seedlings and increasednumber of stunted seedlings culminating in a total failure ofgermination after two years. Slight depletion of food reserves occurred during seed storage.The loss in fat was higher than starch or protein. Total solublesugars decreased while the content of total fatty acids andamino acids and soluble protein increased. Total nitrogen (N)remained unaffected while soluble-N and amino-N increased. Allthese components showed a rapid change (increase or decrease)from 12 months to 18 months of storage which was associatedwith commencement of rapid decline in germinability of the seedsand growth of the seedlings. Initial rapid imbibition of water was observed in viable aswell as non-viable seeds, though at a higher rate in the latterand followed by a lag period in both. At the end of 24 h ofimbibition, water content in non-viable seeds was less thanthat in viable ones. Key words: Voandzeia subterranea, Seed germination, Seed storage     

Characterisation of Soybean and Wheat Seeds by Nuclear Magnetic Resonance Spectroscopy

The effects of equilibration under different air relative humidities (RH, 1 – 90 %) and temperatures (35 and 45 °C) on soybean (Glycine max) and wheat (Triticum aestivum) seeds were studied using different techniques. Seed moisture content, electrical conductivity (EC) of seed leachate and per cent seed germination were measured following standard procedures, and compared with nuclear magnetic resonance spin-spin relaxation time (T₂) measurements. Moisture contents of soybean and wheat seeds, following the reverse sigmoidal trend, were greater at 35 than at 45 °C at any particular RH. Changes in T₂ were related to the changes in germination percentage and leachate EC of both soybean and wheat seeds. Equilibrating soybean seeds at RH 11 % decreased germination percentage with corresponding decrease in T₂. On the contrary, EC of seed leachate increased. In wheat seeds equilibrated at 45 °C, T² was maximal at RH 5.5 %. T₂ declined in seeds equilibrated at high RH (> 80 %) together with low germination percentage.     

Effects of Moisture Content on Germination of Seeds of Hancornia speciosa Gom. (Apocynaceae)

Seeds of Hancornia speciosa germinated best at a temperatureof 20–30 °C. The viability of the seeds during storagewas short and the best storage conditions for viability entailedkeeping the seeds in polyethylene bags. Seed viability was maintainedonly when the seeds were stored at a moisture content above30%; storage conditions which allowed dehydration resulted ina rapid loss of viability (the seeds showed recalcitrant behaviour). Low temperature during storage did not improve longevity. Arelationship between germination and moisture content was established,but when the moisture content fell below 25% there was a drasticreduction of germination. After 9 weeks of storage, even athigh moisture content, seeds lost viability. Loss of seed viability during seed dehydration was associatedwith increased leakage of electrolytes and organic solutes,and reduced tetrazolium staining during subsequent imbibition. Hancornia speciosa, germination, recalcitrant seeds, storage, moisture     

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     

Possible involvement of volatile compounds in the after-ripening of cocklebur seeds

The mechanism of emergence from primary dormancy, the process of after-ripening, in cocklebur (Xanthium pennsylvanicum) seeds was examined in relation to the involvement of volatile compounds and to the relative humidity (RH) in which the seeds were stored. The after-ripening of these seeds proceeds only at water contents between 7 and 14% which are conditioned under RHs of 33% to 53% and are identified with water-binding region II. After-ripening of cocklebur seeds occurred even in water-binding region I. imposed by 12% RH. when exposed to HCN gas during the storage period. Exposure of dormant seeds to acetaldehyde (ethanal) retarded after-ripening. even in water-binding region II. thus decreasing germinability. This decrease of germinability by ethanal was found also in the after-ripened seeds, suggesting that ethanal accelerates seed deterioration rather than retarding the after-ripening. The contents of ethanal. ethanal and HCN were high only in the dormant seeds held at 12% RH. Regardless of RH. a possible conversion of ethanal to ethanol. perhaps via alcohol dehydrogenase. was far larger in dormant than in non-dormant seeds. In contrast, the reverse conversion of ethanol to ethanal was more profound in non-dormant seeds. Pre-exposure of both types of seeds to HCN reduced the contents of both ethanal and ethanol at 12% RH. The contents of various adenylales including ATP in seed tissues were higher in dormant seeds stored at 12% RH than in non-dormant seeds after-ripened at 44% RH. It is suggested that emergence of cocklebur seeds from primary dormancy by HCN treatment at 12% RH may result from the reduction in the contents of ethanal via an unknown mechanism incurring the consumption of ATP. This implies involvement of volatile compound metabolism at the water-binding region II in the after-ripening process of cocklebur seeds.     

Soaking injury and its reversal with polyethylene glycol in relation to respiratory metabolism in high and low vigor soybean seeds

Germination and growth of unaged (high vigor) and accelerated aged (41°C and 100% RH for 4 days; low vigor) soybean [ Glycine max (L.) Merr. cv. Essex] seeds were severely reduced by soaking the seeds under water at 25°C during the first 1 to 8 h of imbibition. Soaking in 30% polyethylene glycol (PEG) did not injure high vigor seeds and improved the performance of low vigor seeds. Soaking in water increased subsequent RQ values and acetaldehyde and ethanol levels. In high vigor seeds, the increase in RQ lasted only 1 h after transfer to aerobic conditions, whereas in low vigor seeds the increase persisted into the second h. Increases in ethanol and acetaldehyde after soaking were more pronounced in low than in high vigor seeds. The data indicate that relatively short periods under water can cause water uptake injury to imbibing soybean seeds and that this injury can be avoided by osmotically reducing the initial rate of water uptake. Seeds whose vigor has declined during accelerated aging show an additional injury which increases with soaking duration and involves alterations in respiratory metabolism including accumulation of acetaldehyde and ethanol to high, and possibly toxic, levels during imbibition.     

Seed Maturity and the Effects of Different Drying Conditions on Desiccation Tolerance and Seed Longevity in Foxglove (Digitalis purpurea L.)

The effects of different drying methods on desiccation toleranceand longevity in seeds of foxglove (Digitalis purpurea L.) wereassessed from just prior to mass maturity (when seeds have attainedmaximum dry weight), and at intervals during the post-abscissionphase of development. Tolerance of drying under seed conservationconditions (15% relative humidity, RH, and 15 °C), was acquiredclose to mass maturity at 36 d after flowering (DAF). Increasesin desiccation tolerance were induced when drying was delayedfor 4 d by placing seeds in a near-saturated atmosphere (approx.100% RH), or if seeds were pre-dried for 7 d at either approx.32% or approx. 73% RH. Irrespective of the drying treatment, seed longevity increasedthroughout the sampling period, i.e. beyond the point of massmaturity and throughout the post-abscission phase, up to thepoint of incipient natural dispersal. At each developmentalstage, delayed drying or pre-drying led to an increase in seedlongevity under controlled ageing conditions compared with seedsdried directly under seed conservation conditions. Increasesin longevity were apparent as increases in the estimates forthe intercept of transformed seed survival curves (K_i) and forthe standard deviation of the normal distribution of seed lifespans,and also in the mean time to death of individuals in storage,consistent with a continuation of ripening events. The results are discussed in relation to the assessment of seedlongevity and to current post-harvest drying practices for seedsintended for long-term ex-situ conservation.Copyright 1995,1999 Academic Press Digitalis purpurea L., foxglove, seed development, seed drying, seed longevity     

Studies on Prolonged Storage of Beauveria bassiana Conidia: Effects of Temperature and Relative Humidity on Conidial Viability and Virulence against Chickpea Borer,Helicoverpa armigera

Unformulated conidia of Beauveria bassiana were stored at five different temperatures (0°, 10°, 20°, 30° and 40°C) at six different relative humidities (RH) (0, 33, 53, 75, 85 and 98%). Conidial viabilities and virulence against third instar larvae of Helicoverpa armigera were determined over a 24‐month period. Conidia survived longest at lower temperatures (0–20°C) and lower RH levels (0–53% RH). At higher temperatures (30–40°C) conidia did not survive. When the temperature was decreased from 30°C to 0°C, at nearly all RH levels the longevity of conidia increased. Conidia remained virulent for third instar larvae of H. armigera under favourable storage conditions for 24 months.     

Factors influencing the storability of Fagus sylvatica L. seeds after release from dormancy

Seeds of beech (Fagus sylvatica L.) that have been subjected to dormancy breaking consisting of 10 weeks of prechilling at 3 °C and 34 % water content (WC) and then desiccation to 10 % WC, are non-dormant (ND). ND seeds are characterised by greater sensitivity to storage conditions, than no prechilled, dormant (D) seeds. The aim of the present work was to investigate factors affecting the loss of seed viability during storage of D and ND beech seeds at different temperatures (4 and 20 °C) and humidity levels (45 and 75 % RH) for 3 weeks. In general, both D and ND seeds maintained a high germination capacity after storage at 4 °C. At 20 °C and 45 and 75 % RH the germination capacity of D seeds diminished to 80 and 28 %, respectively. Under the same conditions, ND seeds lost germination capacity to a greater degree, with only 62 and 7 % germinated seeds, respectively. At 20 °C, an increase in production of reactive oxygen species was observed, and the increase was significantly higher in ND seeds. The loss of germination capacity was coincident with an increase in electrolyte leakage and accumulation of free fatty acids, which suggests that membrane deterioration was the cause of the decline in germinability. ND seeds stored at 20 °C and 45 and 75 % RH showed a greater decrease than D seeds in contents of the primary phospholipids phosphatidylcholine (PC) and phosphatidylethanolamine (PE) as well as in polyunsaturated fatty acids (18:2 and 18:3). ND seeds possessed more unsaturated fatty acids, especially 18:3, than D seeds in the phospholipid fraction before storage. D seeds were characterised by a significantly higher level of α-tocopherol and UV-absorbing phenols. The level of ascorbate was similar in D and ND seeds. D seeds contained glutathione in both reduced (GSH) and oxidised (GSSG) forms, and GSSG dominated GSH. ND seeds contained more GSSG form than D seeds. We concluded that the membranes of ND seeds are exposed to greater oxidative stress during storage due to higher levels of unsaturation and lower levels of α-tocopherol, the main antioxidant that protects membranes against free radical attack.     

浙江早稻谷中稻水象甲的存活研究

研究了早稻收获和储藏期间稻水象甲Lissorhoptrus oryzophilus Kuschel一代成虫的数量动态及其存活时间与温湿度的关系。早稻收割时尚未迁离稻田的象甲中,仅3～13%的个体随脱落操作落入稻谷中,且这些个体经太阳曝晒2天后存活率不足1%;收割时残留于稻草中的虫量为稻谷中的5.1～12.3倍。储藏1个月的稻谷中所有成虫均死亡,但储藏相同时间的秕谷和稻草中存活成虫分别达到250头/50 kg秕谷和17头/50 kg稻草。成虫在≥90% RH下的存活时间显著长于35～75%RH下,在玻璃管中的存活时间显著长于稻谷中。在稻谷中,温度对成虫存活时间的影响不明显,但在玻璃管中,20℃（恒温）下的存活时间显著长于27.0～33.2℃（室温）下。利用生存分析的指数模型分析了成虫存活与湿度、温度和存放基质的关系,表明在75～98% RH之间,湿度越低则成虫死亡风险越大,较高温度（27.0～33.2℃）和处于稻谷中其死亡风险亦高。这些结果为开展稻谷中稻水象甲的检疫提供了依据。     

Some Morphological and Chemical Characteristics of Developing Fruits of Raphia hookeri

Ndon, B. A. 1985. Some morphological and chemical characteristicsof developing fruits of Raphia hookeri.—J. exp. Bot. 36:1817–1830. Fruits which were at different stages of development were randomlysampled from different inflorescences of Raphia hookeri palms.The morphological characteristics and chemical (the dry matter,lipid and carbohydrate) contents of the exocarp and seeds weredetermined. The results showed that the seed length, circumferenceand volume were optimal at 24 months after pollination whichindicates that Raphia seeds attained maximum size at that period.The seed endosperm was liquid or semi-liquid between 6–18months after pollination but became solid with a prominent embryoat 24 months. The seed dry matter was low at the early stagesof development but there was a rapid increase in seed dry weightat 18–33 months after pollination. The seeds were physiologicallymatured at 30–33 months after pollination, while the exocarpmatured at 24–30 months after pollination. The Raphia seeds were low in lipid (about 2%) compared to theexocarp which had 30–40% lipid at full maturity. Maximumamount of lipid was accumulated within the exocarp at 36–42months after pollination and this period indicates the timefor harvesting Raphia fruits for maximum oil which is probablythe most economic part of the fruit. The total sugar concentration increased in the exocarp withincrease in maturity. Conversely the concentration of sugarsdecreased within the seeds as the fruit matured. Maximum totalsugar concentration (about 309 mg g^–1 dry fat free sample)was found in the exocarp at 36–42 months after pollination.Mature seeds at 48 months after pollination had about 50 mgof total sugars per g of fat free sample. There was insignificantaccumulation of starch in the exocarp. The mature seeds werelow in starch (5–10% of the dry weight). Key words: Raphia hookeri, development, fruit     

Use of Coating to Protect Lyophilized Bacillus popilliae from Moisture

Lyophilized cells of Bacillus popilliae were protected from moisture when suspended in pellets of tung oil polymer which were then coated with paraffin wax. The survival of the protected cells at various levels of relative humidity (RH) and under various storage conditions was determined. During 6 months of storage, moisture appeared to have little effect on survival of the cells when the RH level was 22% or less; but, at higher RH levels, survival declined upon storage. Viable cells were recovered when pellets were stored for 3 months at 33% RH, 2 months at 42% RH, 1 month at 50% RH, and 4 days in distilled water. Under field conditions, some cells survived at least 1 week of storage.     

Temperature Dependence of Viability and Dormancy of Zizania palustris var. interior Seeds Stored at High Moisture Contents

Seeds (caryopses) of North American wild rice (Zizania palustrisvar. interior), a temperate aquatic grass, have been thoughtto require storage at low temperatures and high moisture contentsto preserve viability. The seeds are also deeply dormant atmaturity and require up to 6 months of stratification to breakdormancy. We report here that wild rice seeds can retain viabilityat moisture contents 30% (f. wt. basis) for up to 6 monthsat temperatures as high as 30 °C, and for at least 1 yearat temperatures below 20 °C. Dormancy is not broken at temperaturesabove 10 °C, but subsequent stratification requirementsare unaffected by prior warm storage. Cold storage is thereforenot required to maintain viability of wild rice seeds, but isnecessary to break dormancy. Hydrated wild rice seeds can befrozen to –10 °C without damage, but dormancy is notlost at subfreezing temperatures. These results provide newoptions for long-term storage of wild rice seeds. Zizania palustris var. interior (Fassett) Dore, wild rice, seed, germination, dormancy, storage, moisture content     

An Intermediate Category of Seed Storage Behaviour?: I. COFFEE

Seeds of four cultivars of arabica coffee (Coffea arabica L.)were tested for germination following hermetic storage for upto 12 months at several different combinations of temperaturesbetween –20 °C and 15 °C and moisture contentsbetween 5% and 10% (wet basis). Most of the seeds from one cultivarwithstood desiccation to between 5% and 6% moisture content,a seed water potential of approximately –250 MPa, butthose of the remaining three cultivars were much more sensitiveto desiccation damage. Moreover, in all four cultivars, seedlongevity at cool and sub-zero temperatures, and at low moisturecontents did not conform with orthodox seed storage behaviour:viability was lost more rapidly under these conditions thanat either warmer temperatures or higher moisture contents. Theresults confirm that coffee seeds fail to satisfy the definitionsof either typical orthodox or recalcitrant seed storage behaviour.These results, therefore, point to the possibility of a thirdcategory of storage behaviour intermediate between those oforthodox and recalcitrant seeds. One of the main features ofthis category is that dry seeds are injured by low temperatures. Key words: coffee, Coffea arabica L., seed storage, seed longevity, desiccation, temperature     

Development of the Major Triterpenoids and Oil in the Fruit and Seeds of Neem (Azadirachta indica)

In order to learn the best time for harvesting Neem (Azadirachtaindica) seeds, the amount of the five major triterpenoids, togetherwith the oil content have been determined throughout a fruitingseason in six selected trees in Sri Lanka. The triterpenoidcontent and the relative proportions of the major compoundschanged little from the hard green fruit stage to mature seeds,while the oil content increased markedly with time. The highestcontent of azadirachtin (10 mg g^-1seed kernels) was recordedin newly ripened seeds. There was some loss of salannin andazadirachtin in storage after harvesting for up to 6 months. Azadirachtin; salannin; nimbin; neem seeds; neem fruit; seasonal change; neem oil; seed storage; Azadirachta indica ; Meliaceae     

Ageing of tomato seeds involves glutathione oxidation

The effect of seed ageing on the oxidation of reduced glutathione (GSH) and the role of GSH oxidation in ageing-induced deterioration were studied in seeds of tomato ( Lycopersicon esculentum Mill. cv. Lerica, Moneymaker and Cromco). Both long-term storage at 15°C/30% relative humidity (RH) and artificial ageing at 20°C/75% RH, 30°C/45% RH and 60°C/45% RH resulted in a marked loss of GSH and a simultaneous, though not proportional, increase in its oxidized form GSSG. The glutathione thiol-disulfide status shifted towards a highly oxidized form, while the total glutathione pool decreased. The extent of GSH oxidation differed between ageing conditions and was not directly related to the extent of seed deterioration. Thiobarbituric acid-reactive substances did not increase in ageing tomato seeds, suggesting that lipid peroxidation did not take place. Hydration of seeds, either upon imbibition in water or by priming in an osmotic solution, resulted in a rapid decrease in GSSG, a shift of the glutathione redox couple to a mainly reduced status and an increase in the glutathione pool, in both control and aged seeds. The results indicate that, in tomato seeds, (1) seed ageing involves GSH oxidation into GSSG, which is indicative of oxidative stress, (2) ageing does not affect the GSSG reduction capacity upon subsequent imbibition, and (3) the lowered viability of aged seeds cannot directly be ascribed to the decreased GSH pool or To the highly oxidized glutathione redox status.     

Effect of Different Seed Storage Conditions on Germination and Isozyme Activity in Some Brassica Species

Sixteen seed accessions of four Brassica species (B. cretica,B. incana, B. montana and B. oleracea) were examined following5-22 years' storage in the germplasm bank. Germinability andisozyme of seeds stored under long-term (-10 °C and 3% moisturecontent) and short-term (5% °C and 8% moisture content)storage conditions were compared. Long-term storage producedno deterioration and the ability to germinate was satisfactorilymaintained over 8-22 years. Short-term storage conditions maintainedgermination ability up to 10-12 years in all accessions of B.cretica and B. montana. However, seed multiplication might beessential every 10 years for some accessions of B. oleraceastored in this way. In the 8 to 9-year-old accessions of B.cretica, B. incana and B. montana, no significant differenceswere detected between conservation systems for germinabilityand frequency of seeds showing isozyme activity for seven enzymesystems (ACO, IDH, MDH, ME, PGI, PGM and 6-PGD). However, significantdifferences were found for the ADH enzyme system. Moreover,in the 5-22-year-old B. oleracea accessions, significant differenceswere found between storage conditions with respect to isozymeactivity for all enzyme systems studied.Copyright 1995, 1999Academic Press Brassica cretica, Brassica incana, Brassica montana, Brassica oleracea, accessions, isozymes, germination, seed storage     

Seed Dormancy in Acer: The Role of Testa-imposed and Embryo Dormancy in Acer velutinum

The embryo dormancy shown in freshly harvested samples of Acervelutinum seeds is weakly established and very short-lived.Loss of this embryo dormancy occurred during post-harvest fruitstorage at either 5 or 17 C. In contrast, the dormancy of intactfruits and seeds was overcome only during storage at the lowertemperature. Removal of the cotyledons from embryos of freshlyharvested fruits allowed more rapid germination of the embryonicaxes, indicating that the cotyledons exert an inhibitory effect,although the axes still retained a measure of innate dormancy.The inhibitory effect of the cotyledons became less marked withincreasing duration of fruit storage, this loss of inhibitoryeffect occurring at both storage temperatures. Applied ABA stronglysuppressed germinative capacity in intact embryos and isolatedembryonic axes from freshly harvested fruits, but when ABA wasapplied to embryos of fruits that had been stored for variousperiods at 5 or 17 C, the inhibitory effect was first weakenedand then lost with increased storage. Although dormancy in the seeds of A. velutinum may be describedas intermediate between testa-imposed dormancy and true dormancy,it is perhaps more properly included in the former category. Acer velutinum Boiss. var. vanvolxemii, abscisic acid, embryo dormancy, germination, seed storage, testa-imposed dormancy, tissue sensitivity     

Survival of Lyophilized Bacillus popilliae in Soil

Survival of lyophilized vegetative Bacillus popilliae in dry soil and in soil kept in atmospheres with different relative humidities (RH) was determined. Cells survived for at least 1 year when the RH was 22% or lower, but an RH of 33% or higher reduced viability. At 33% RH, no bacteria were recovered after 5 months of storage; at 42% or above, no bacteria were recovered after 1 month of storage.