Lipid participation in intracellular freezing avoidance mechanisms of lettuce seed

Freeze-fracture electron microscopy was used to study water content related freezing resistance in Grand Rapids lettuce seeds. Consistent and recognizable conformational changes occurred in lipid-water phases of lettuce seeds at different moisture contents. In air-dry lettuce seed cotyledons, the lipids lying in spherical lipid bodies near the cell wall appeared amorphous, while the structure was crystalline above 20% water content. The lipid bodies interassociated into membrane bilayers in seeds containing 20 to 25% water. Such lyotropic phase transitions in membrane lipids during lettuce seed hydration are believed to contribute to the biphasic freezing behavior observed in lettuce seeds at different moisture contents and to provide a natural freezing tolerance mechanism for highly desiccated plant tissues such as seeds.     

Seed Dormancy,Germination and Storage Behavior of Magnolia wilsonii (Magnoliaceae), an Endangered Plant in China

In this paper, we measured the embryo morphology and size of Magnolia wilsonii seeds. And the effects of temperature, stratification, KNO3 and soaking on seed dormancy and germination were studied. Seeds were desiccated to different moisture content, and then stored at 4℃ and -20℃ for 100 d. The effects of desiccation and storage temperature on seed viability of M. wilsonii were analyzed. The results showed that the underdeveloped embryos of Mwilsonii seeds needed cold stratification to achieve physiological afterripening. The favorable temperature for germination of Mwilsonii seeds was 25/20℃. Cold stratification and GA3 could effectively overcome dormancy of Mwilsonii seeds. Therefore, Mwilsonii seeds have morphophysiological dormancy. About 5350% of seeds could survive in the moisture content of 539%. After 100 d storage at -20℃, all seeds in different moisture contents died. However, after 100 d storage at 4℃, 76% of seeds survived. Thus, Mwilsonii seeds exhibited intermediate seed storage behavior. The optimum storage environments was dry or moist storage at 4℃.     

濒危植物西康玉兰种子休眠、萌发及贮藏特性

研究了西康玉兰(Magnolia wilsonii)种胚的形态大小,温度、层积、硝酸钾、浸种处理和GA_3对种子休眠及萌发的影响.通过将种子脱水至不同含水量和在两种温度(4℃和一20℃)下贮藏100 d后测其生活力,分析各处理对西康玉兰种子活力的影响.结果表明:西康玉兰种子胚尚未分化完全,需经过低温层积完成生理后熟;低温层积和GA3可打破种子休眠.种子最适萌发温度为25/20℃.由此推测,西康玉兰种子具有形态生理休眠特性.西康玉兰种子当含水量降至5.39%,存活率为53.50%.不同含水量的种子贮藏在-20℃条件下,100 d后种子全部死亡;但在4℃下含水量为10.31%的种子存活力高达76%.因此,西康玉兰种子极可能属于中间性种子,其适宜贮藏环境为4℃下的干藏或湿藏.     

The Potential for Newly-Germinated Cabbage Seed Survival and Storage at Sub-Zero Temperatures

The moisture content of newly germinated cabbage seed (radicles1 05 mm long) was reduced to 14% of f.wt without loss of viability.As the moisture content was reduced below 45%, the temperatureat which the germinated seeds froze, and therefore died, decreasedprogressively to a minimum of –34 C at 19% moisture content.No freezing exotherms were recorded in seeds with moisture contentsbelow 19%. Seeds with a moisture content between 14 and 16%maintained viability for at least 1 week when cooled at 26C.min^–1to –20 C and held at this temperature, indicating thepotential for prolonged storage of these low-moisture-contentgerminated (LMCG) seeds. Brassica oleracea, cabbage, germinated seed, seed storage, fluid drilling, freezing exotherm, thermal analysis     

Storage of ethyl methanesulphonate-treated barley seeds with low moisture contents

Barley seeds were treated with ethyl methanesulphonate (EMS) for 3 h at 25° C, washed with tap water for 24 h at 25° C, redried at 40° C to different moisture contents below 15% and stored at 25° C in desiccators or in sealed plastic bags. The criteria used for expressing the effect of storage were the M₁ seedling height and the frequency of chromosomal aberrations. With 14·9% seed moisture a strong increase of biological injury occurred in the course of a 2-week storage, while storage of seeds having an initial moisture content of 11·7% led to a significant increase of injury only after 6 weeks. Superdry EMS-treated seeds with 5% or less moisture can be stored at 25° C without any changes in the biological effects. A method is recommended to avoid the EMS-storage effects.     

Wheat Viability at High Moisture Content under Hermetic and Aerobic Storage Conditions

Wheat seeds (Triticum durum) were stored under both hermeticand aerobic conditions at 25 °C with moisture contents from15 to 33 per cent. Under hermetic storage, seeds lost viabilitymore rapidly the higher the moisture content, whereas in aerobicstorage, seed longevity was enhanced as the moisture contentwas increased from 24 to 31 per cent, and over this range ofmoisture content the seeds survived longer under aerobic thanhermetic storage. On the contrary, an apparent reversal of thistrend occurred when moisture content was increased above 31per cent. The possibility that the changes in longevity occurring at highermoisture contents might be due to the activation of seed metabolismwas supported by the enhanced incorporation of [³H]leucine intoTCA insoluble material (indicating increased protein synthesis)and the reduced leakage of glucose (indicating increased membranerestitution) when seeds were allowed to achieve higher moisturecontents during the prehydration period. The highest level ofseed activation was found in seeds preconditioned to about 31per cent moisture content. Moreover, these seeds, when subsequentlystored under aerobic conditions, maintained a higher rate ofprotein synthesis and lower membrane permeability during thestorage period than seeds at lower water contents. It is suggested that seeds stored at a sufficient hydrationlevel in the presence of oxygen can sustain an effective metabolismduring extended storage, thus permitting the repair of cellulardamage. However, it might be possible that at about 33 per centmoisture content seed could suffer from an excessive advancementof metabolism. Triticum durum, seed storage, effects of high moisture content and oxygen     

顽拗性竹柏种子的贮藏特性

文章对竹柏(Podocarpus nagi)种子的脱水耐性和贮藏特性进行了研究,结果表明:竹柏种子成熟时初始含水量约为(35±0.7)%,种子对脱水敏感,其最低安全含水量约为(16.86±0.73)%,具有顽拗性种子的典型特征;湿藏和半干藏都可以作为短期保存竹柏种子的方法,且以4℃保存优于15℃保存,但不管种子含水量如何,零下低温保存对竹柏种子都是致命的;半干藏法保存实验中,未进行脱水处理的种子(对照)在4℃贮藏6个月,种子萌发率没有发生明显下降,但贮藏期延长到9个月时,临界含水量的种子萌发力保存最高;不管贮藏介质的含水量高低,也无论贮藏在4℃还是15℃,湿藏种子在9个月的贮藏期内萌发率均没有明显的降低,但当贮藏到12个月时,15℃湿藏种子的萌发率显著高于4℃贮藏的种子,但15℃湿藏的种子在贮藏到3个月时即发现种子在贮藏期间萌发,且随着贮藏介质含水量的升高和贮藏期的延长,萌发的种子增多;竹柏的离体胚经过2 h硅胶快速脱水至含水量7%后再冷冻即可获得90%以上的融后存活率,且超低温保存1年的离体胚解冻后,与只保存1周的存活率没有明显差异,表明超低温长期保存竹柏种子是可行的。本研究可以为进一步探究顽拗性种子的短期贮藏和长期保存提供理论基础和基础资料。     

Desiccation and storage studies on Dipterocarpm seeds

The relationship of seed moisture content (fresh weight basis) to germination, and the effect on viability of various storage conditions were examined for five species of the tropical forest tree genus Dipterocarpus. It was shown that seeds fall into two groups with regard to desiccation tolerance. Firstly, D. obtusifolius and D. turbinatus cannot be dried below about 45% moisture content without damage; a sigmoid curve was found to fit the relationship between germination and moisture content for the latter species. Secondly, D. intricatus, D. tuberculatus and D. alatus can be safely dried to 10%, 12% and 17% moisture contents respectively, but desiccation to near 7% moisture content reduced viability by at least a half. Storage studies showed that seed of D. intricatus and D. tuberculatus possessed increased longevity as moisture contents were reduced within the range 6–20%. It was concluded that seeds in the first group are ‘recalcitrant’ and that those in the second group are ‘orthodox’ in their storage physiology, according to the categories described by Roberts (1973). Wide differences between species in seed desiccation rates were observed. In 15% relative humidity D. intricatus dried to 7% moisture content within a week, whilst D. obtusifolius retained 30% moisture content even after 5 wk; other species had intermediate desiccation rates. Seed size and structure may partly account for the differences observed. Correlations were observed between seed storage physiology and other factors which were investigated. ‘Orthodox’ seeds had quicker desiccation rates, were derived from drier habitats, and had smaller embryos than those of ‘recalcitrant’ seeds. ‘Orthodox’ seeds, with the possible exception of D. alatus, should be kept at 0–3°C with about 12% moisture content in the short term and, provided less than 10% germination is lost on freezing, at-18°C with about 8% moisture content in the long term. ‘Recalcitrant’ seeds should be stored in ventilated containers at 21°C and with moisture contents above 45–50%.     

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     

Biological,physical and chemical changes in stored wheat

Six lots of wheat from six farmers' bins in Manitoba were adjusted to a range of 5 moisture contents, held at 6 temperatures and sampled at 6 times during storage. After sampling, seeds were surface sterilized with mercuric chloride and subsamples plated on Czapek's agar, on filter paper moistened with water, or on filter paper moistened with a 7.5% sodium chloride solution. The microflora on the seed was determined, germination counts were made, and the seed was examined by government inspectors for its condition and grade. A total of 1192 samples were examined. In addition, 180 samples were subjected to a fat acidity test. Deterioration for each combination of temperature and moisture, the fungi involved, and the consequent effect on condition, grade and fatty acid content were observed. The effect of high moisture content (>20%) and low temperatures (3–10 ° C) on infection of seed by Penicillium and the consequent effect on germination, condition, grade and fatty acid value are stressed. The interrelationship among fungi, and among fungi and temperature, moisture, storage time, fat acidity values and germination are indicated by correlation coefficient matrices.Deceased July 25, 1982.     

Ergosterol accumulation and oil quality changes in stored soybean invaded by Aspergillus ruber (A. glaucus group)

Soybean seed samples inoculated with spores of Aspergillus ruber were stored for 20 to 140 days at 25 ± 1 °C with moisture content varying from 11.3% to 17.7%. Seeds were colonised by the fungus within 20 days at all moisture levels. Ergosterol concentration in seeds increased with time of storage, being slow in samples with moisture content of 11.3% to 13.1% and more rapidly in those with higher moisture content. Free fatty acid (FFA) content also increased following the pattern of ergosterol. Equations were used to predict minimum safe storage period of the seeds at 25 °C. The rise in FFA is due to increase of A. ruber mass within the seed. Fungal growth did not affect fatty acids profiles or iodine index of the extracted oil. The results suggest revision of the present recommendations regarding seed moisture during storage. The FFA concentration of a seed lot can be used as a sensitive indicator of seed deterioration due to storage fungi. This revised version was published online in June 2006 with corrections to the Cover Date.     

Viability of rape (Brassica napus L.) seeds following selection on the basis of newly-emerged radicles then subsequent drying and storage

Germinating rape seeds selected on the basis of newly-emerged radicles (1 ± 0.5 mm) were dried to an equilibrium moisture content (c. 11%) in air at 20°C and 80% relative humidity without loss of viability. Storage life of these low-moisture-content germinating (LMCG) seeds at 15°C was limited to 7 days before viability was significantly reduced. However, viability of LMCG seeds was maintained for 84 days in storage at -20°C. Longer periods in store reduced viability, but 96% of seeds still remained viable after 336 days at - 20°C. Increasing periods of storage at -20°C reduced the subsequent seed longevity at 15°C, indicating a reduction in vigour during storage. Storage under reduced pressure or in a nitrogen atmosphere had little significant effect on seed longevity. Reduction of moisture content below 11% using vacuum drying at a range of temperatures reduced seed vigour.     

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     

Water Content and the Conversion of Phytochrome Regulation of Lettuce Dormancy

In an effort to determine which biological reactions can occur in relation to the water content of seeds, the regulation of lettuce seed dormancy by red and far red light was determined at various hydration levels. Far red light had an inhibiting effect on germination for seeds at all moisture contents from 4 to 32% water. Germination was progressively stimulated by red light as seed hydration increased from 8 to 15%, and reached a maximum at moisture contents above 18%. Red light was ineffective at moisture contents below 8%. Seeds that had been stimulated by red light and subsequently dried lost the enhanced germinability if stored at moisture contents above 8%. The contrast between the presumed photoconversion of phytochrome far red-absorbing (Pfr) to (Pr) occurring at any moisture content and the reverse reaction occurring only if the seed moisture content is greater than 8% may be explained on the basis of the existence of unstable intermediates in the Pr to Pfr conversion. Our results suggest that the initial photoreaction involved in phytochrome conversion is relatively independent of water content, while the subsequent partial reactions become increasingly facilitated as water content increases from 8 to 18%.     

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     

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     

温度和水分及盐分胁迫对银沙槐种子萌发的影响

利用控制实验研究了温度、湿度、干旱和盐分胁迫等生态因子对银沙槐种子萌发的影响,以探索银沙槐种子对各种生态因子的适应性。结果显示:(1)银沙槐种子在20℃、25℃恒温和15℃/25℃、20℃/30℃、10℃/20℃变温环境中的发芽率较高且无显著差异,其在20℃恒温、15℃/25℃、20℃/30℃变温条件下的发芽指数较高,但差异不显著。(2)土壤含水量在1%～5%之间,各水分处理间种子发芽率差异显著(P<0.05),而在5%～25%间种子发芽率变化不显著。(3)盐胁迫和水分胁迫对银沙槐种子的萌发均有明显的抑制作用,可显著降低种子萌发率(P<0.05);种子发芽指数和活力指数均随渗透势和NaCl浓度增大而显著减小(P<0.05);恢复萌发率随渗透势和NaCl浓度的增加而显著增加(P<0.05)。研究发现,银沙槐种子萌发最适温度为20℃恒温和15℃/25℃变温,最适土壤含水量为10%～25%;种子萌发对盐分和干旱胁迫表现出不同程度的耐受性,萌发过程中主导抑制因素为渗透胁迫,离子毒害作用甚微;银沙槐种子休眠机制和萌发特征表现出它对生境的良好适应性。     

濒危植物多毛坡垒种子萌发的生理生态特性

研究了多毛坡垒（Hopea mollissima）种子在自然条件下的寿命及成熟度、温度、光照、土壤水分条件对多毛坡垒种子萌发的影响,结果表明：多毛坡垒种子适宜萌发的温度范围是20～30℃,最适宜温度为25℃;光照对多毛坡垒种子的萌发没有显著性影响,但在周期性光照条件下以25～30℃萌发结果最好,全黑暗条件下以20～25℃萌发结果最好;种子适宜萌发的土壤含水量为20％～60％,最适含水量为50％～60％;种子成熟后在树上长时间宿存明显降低了种子的发芽率和活力;在室内自然摊放条件下,10天后发芽率减半,16天后完全丧失发芽能力。研究认为,该物种对热带湿性季节性雨林原生境有很强的依赖性,目前多毛坡牟的濒危状态主要是由于滥砍乱伐和森林破坏造成的,由于其种子具有顽拗性种子的一些特点,日前该物种宜采取活体保仔的方法,以就地保护为主,活植物迁地保护为辅。     

Changes in soluble amino nitrogen,protein, nitrate reductase activity and abscisic acid during development of wheat grain

Trends in the time course, changes in the moisture, soluble amino acids, proline, abscisic acid contents and nitrate reductase activity determined byin vivo method in the developing seeds of wheat were studied. Maximum dry matter augmentation in the seed took place in the period between 10–30 days after anthesis. Per cent moisture and moisture content started declining 15 days and 25 days after anthesis, respectively. Levels of soluble amino acids, proline and nitrate reductase activity were higher during initial stages of seed development, but decreased with increasing magnitude of dehydration and accumulation of abscisic acid (ABA) in the maturing seeds.     

Viability of Lettuce Seeds: II. SURVIVAL AND OXYGEN UPTAKE IN OSMOTICALLY CONTROLLED STORAGE

Lettuce seeds were stored under aerobic conditions at varioustemperatures from 20 ?C to 35 ?C, under anaerobic conditionsat 35 ?C, and at various moisture contents ranging from 17%to 44%, controlled osmotically by polyethyleneglycol. Survivalof these seeds was compared with others maintained over a similarrange of temperatures and moisture contents but in which themoisture contents were controlled by hermetic storage. Irrespectiveof the method of controlling moisture content, under anaerobicconditions the trend (shown previously in drier seeds) of decreasein longevity with increase in hydration continued up to about27% moisture content, but above this value there was no furtherchange in longevity. In contrast, under aerobic conditions longevityincreased exponentially up to 44% moisture content, this increasestarting at about 15% moisture content at 20 ?C and at about24% at 35 ?C. Under all circumstances longevity was less athigher temperatures. On a water substrate, thermodormant seeds showed a rapid increasein rate of oxygen uptake, followed by a rapid decline, and thena slower decline to a steady state. Similar phases were shownin seeds of various moisture contents controlled osmoticallydown to 17%, although the corresponding rates were lower withdecrease in moisture content and temperature, and the phaseswere extended at lower temperatures. The implications of these results are discussed in relationto the conditions needed to sustain sub-cellular repair andturnover as postulated by Villiers. Key words: Lettuce, Lactuca sativa L, Seed viability, Seed storage, Respiration