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     

生菜种子低温处理后耐冻性和脂肪酸合成代谢的关系研究

以生菜(Lactuca sativa)种子为研究对象,通过不同时间的吸水处理分析其含水量变化,再通过程序降温处理,分析不同含水量种子发芽率的差异,以及脂肪酸合成有关基因(FAD2、FAD3、PPT、ELOVL)和冷调节基因ICE1的表达。结果表明,种子含水量随吸水时间增加而升高。程序降温至同样的低温冷冻条件下(-20℃、-22℃),吸水时间小于6 h的种子发芽率较高,而吸水8 h以上的种子发芽率显著降低。种子吸水8 h含水量处于饱和状态,在此状态下种子对低温较为敏感,说明含水量对种子耐冻性有影响。冷冻处理后生菜种子基因表达检测结果表明,脂肪酸去饱和酶基因(FAD2、FAD3)、蛋白质棕榈酰基硫脂酶相关基因(PPT）、长链脂肪酸延伸酶相关基因(ELOVL)的表达水平均随着种子含水量增加呈上升趋势,吸胀10 h的种子表达量最高,此时种子由于高含水量所受冷冻伤害最大。基因ICE1在冷冻处理种子中的表达也随着吸水时间增加而升高,在吸水10 h时种子中表达量到最高水平。综上,种子含水量越高,所受冷冻伤害越大。但种子在低温条件下具有一定的抗冷反应,可通过相关基因的过表达调控合成更多不饱和脂肪酸、抗冻蛋白等提高含水种子耐冻性。     

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     

Effect of desiccation on physiological and biochemical indicators associated with the germination and vigor of cryopreserved seeds of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L. cv. Sancti Spíritus 96

Cryopreservation is a valuable technique for the long-term conservation of plant germplasm and complementary to traditional seed storage methods. However, critical factors such as seed moisture content should be optimized before using this technique as a safe strategy for storing seeds such as those of Nicotiana spp. This study aimed to determine the effect of desiccation on physiological and biochemical indicators associated with germination and vigor in cryopreserved seeds of Nicotiana tabacum cv. Sancti Spíritus 96 (SS-96). The germination and vigor of seeds with a range of moisture content were assessed using electrolyte leakage and accelerated aging tests. In addition, these physiological indicators were related to the oxidative state of the seeds, in terms of the rate of O₂ ^·? generation and the H₂O₂ content, and the activity of enzymatic antioxidants superoxide dismutase and catalase. The cryopreserved seeds of N. tabacum SS-96 with a moisture content of 2.1% exhibited higher vigor probably due to the retention of membrane integrity, reflected by lower levels of lipid peroxidation and electrolyte leakage associated with the absence of oxidative stress. The results suggest 2.1% as the optimal moisture content for the storage of seeds of this cultivar, both at cryogenic temperatures and at 5°C.     

Storage behavior of Zygophyllum xanthoxylon (Bge.) Maxim seeds at low moisture contents

This research aimed to determine whether low moisture content storage improves the longevity and storability of Zygophyllum xanthoxylon (Bge.) Maxim seeds. The seeds of Z. xanthoxylon were dried from 11.43 to 4.81%, 3.81 and 2.41% moisture content in a desiccating container with silica gel. After storage for 24 months their level of vigor was measured. After the ageing process, some physiology indices, including the relative electrical conductivity, SOD, PER, CAT, volatile aldehydes and malondialdehyde were tested. The results indicated that SOD, PER and CAT activities of the low moisture content seeds were higher than the control seeds while volatile aldehydes and malondialdehyde were lower than the control group. The electrical conductivity showed that there was obvious statistical difference between low moisture content seeds with the control group. All the results showed that low moisture content storage is beneficial for maintaining the vigor of Z. xanthoxylon seeds. Thus Z. xanthoxylon seeds can be stored with low moisture content at ambient temperature condition for a long time.     

Studies on Desiccation and Storage of Mango Seeds

Mango (Mangifera indica L.) seed is recalitrant which taken from ripened fruits con,ained as high as 69.2'–75.5% moisture content (The moisture content of embryonic axis is 73.8–86.3%). When seeds were naturally dried for 8 days, the moisture content declined to 39.1% (in embryonic axis the moisture content declined to 46.5%) and the viability of seeds completely lost. Embryonic axis lost water slower than whole seed because of the prevention of desiccation by the large cotyledons. During natural desiccation, the conductivity of leachate increased rapidly from 2.2 μΩ·cm-1·g-1 (the same unit below) to 56.7, whereas the activities of dehydrogenase and acid phosphatase decreased drastically. When seeds were rapidly dried for 42 hours, the moisture content declined from 75.5 to 29.9%, the conductivity of leachate increased from 1.8 to 36.9 and the percentage germination changed from 100% to 10%. Desiccation damaged the cell membrane and decreased the activities of enzymes. Rapid drying was better for maintaining longevity than natural (slow) drying because the former did less damage to the cell membrane than natural (slow) drying as shown in the conductivity changes. The moisture content of excised embryonic axis decreased to 11.8% when they were dried for 8 hours by silicagel. The survival percentage of these embryonic axis was 80% when they were incubated in MS+0.2 mg/L BA+2.0 mg/L NAA+500 mg/L gln+3% sucrose+0–9% agar medium. Seeds with 51.0% moisture content (rapidly dried for l0 hours by electric fan blowing) had 65% viability after 7 months wet storage with the polyethylene bag at 15℃.     

Dormancy Breaking and Storage Behavior of Garcinia cowa Roxb. (Guttiferae) Seeds: Implications for Ecological Function and Germplasm Conservation

The dormancy breaking and storage behavior of Garcinia cowa Roxb. seeds were investigated.The seeds of G. cowa had 8-11 months dormancy in their natural habitat. Seeds were matured and dispersed at the end of the rainy season (mid-late August to late September) and were scatter-hoarded by rodents as food for winter after the seeds had fallen to the ground. Seedlings often emerged in the forest during the rainy season (May to August) the following year. Intact seeds of G. cowa failed to germinate after being sown at 30 ℃ for 120 d and the mean germination time (MGT) of seeds cultured in a shade (50% sunlight)nursery was 252 d. The most effective method of breaking dormancy was to remove the seed coat totally,which reduced the MGT to 13 d at 30 ℃. Germination was also promoted by partial removal of the seed coat (excising the hilum and exposing the radicle) and chemical scarification (immersion in 1% H2O2 for 1 d).Unscarified seeds take up water rapidly in the first 96 h, but water was absorbed by the outside seed coat,without penetrating through it. The moisture content (MC) of G. cowa seeds was high (50% in fresh weight)at shedding. The seeds could tolerate desiccation to some extent, until the MC reached approximately 40%;below that, the viability decreases rapidly and all seeds died at approximately 17% of MC. Seed viability decreased rapidly when seeds were chilled at 4 ℃; germination was 2% after storage for 1 week. Even stored at 10 ℃, seeds began to be damaged after 4 weeks. Seed storage for 1 yr revealed that in both dry (relative humidity (35 ± 5)%) and moist (wet sand) storage conditions, seed viability declined, but germination percentages for seeds stored under moist conditions are better than for seed stored under dry conditions.Because of their low tolerance to desiccation, marked chilling sensitivity and relatively short lifespan, G.cowa seeds should be classified into the tropical recalcitrant category. The ecological implications of dormant recalcitrant seeds and cues on storing recalcitrant seeds were discussed.     

Desiccation Intolerance in Seeds of Zizania palustris is Not Related to Developmental Age or the Duration of Post-harvest Storage

Freshly harvested seeds of Zizania palustris L. representingfour distinct stages of development were intolerant of desiccation.Rapid drying did not improve survival. The relationship betweenprobit viability and embryo moisture content for freshly harvestedmature seeds was identical to that described in an earlier publicationfor seeds previously stored fully imbibed at low temperaturesto remove dormancy. A small reduction in embryo moisture contentduring the first 3 d of drying at 15% r.h. and 15°C ledto a highly sigificant (P < 0.01) increase in the proportionof fresh seeds capable of germination in response to a rangeof dormancy breaking factors including giberellins. Resultsare discussed in relation to previous work on recalcitrant seedsand to the widely reported developmental changes in orthodoxseeds which are triggered by desiccation. Zizania palustris, wild rice, seeds, drying, desiccation intolerance, development     

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 fumigation of seeds with methyl bromide

Forty varieties of vegetable, cereal, fodder and grass seeds at 10, 13 and 16% moisture content were fumigated with methyl bromide at 10 and 18^oC, to determine the maximum safe moisture content required to avoid unacceptable phytotoxicity. Dosages (concentration x time products) of 200 and 400 mg h/1 of technical and commercial methyl bromide were used and treated seeds were compared with controls in germination tests. Results were not consistent, but in c. 30 % of the comparisons lower moisture contents, lower temperature and lower dosage each resulted in better germination than the corresponding higher values. The reductions in germination of treated samples were usually less than 5 % and the overall average reduction was only 2.3%. This work showed that under controlled conditions the treatments are safe for most of the tested varieties of seed, but for some seeds, particularly white clovers, Agrostis and runner beans, the highest moisture content (16%) should be avoided.     

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.     

A study of the optimum drying conditions for cabbage seed following selection on the basis of a newly-emerged radicle

A series of experiments were carried out to study the effects of different drying regimes on the survival of newly-germinated (radicle emerged 0·5-1·5 mm through the seed coat) cabbage seeds and to establish a treatment for reducing the moisture content of these germinated seeds. A preliminary surface-drying step proved necessary to avoid seed agglutination during the main drying stage. Of the drying regimes compared both viability and vigour were most effectively maintained in air, conditioned to between 20 and 30°C and with 80% r.h., flowing through the seeds at 0·25 m s^-1. Under these conditions, the viability and vigour of newly-germinated seed was maintained during drying to an equilibrium moisture content of c. 14%. At this moisture content further radicle growth was prevented and seeds remained free-flowing and separate for sowing or for further treatment.     

沙芥种子发育过程中的脱水耐性

以发育过程中经脱水和未脱水处理的沙芥种子为试验材料,测定了其含水量、萌发率和抗氧化酶系统,探讨了沙芥种子脱水耐性与抗氧化系统之间的关系。结果表明：在20～60DAF,沙芥种子含水量逐渐下降,干重逐渐增加;60DAF种子具有萌发能力,萌发率为24％;且脱水可促进沙芥种子的萌发,人工脱水至含水量为12％和5％时,萌发率分别为56％和44％,自然脱水至含水量为12％和5％时,萌发率分别为52％和60％;发育过程中沙芥种子SOD活性逐渐降低,而CAT、POD、LOX活性以及MDA含量均呈上升趋势;在脱水过程中,随着种子含水量的下降SOD活性逐渐降低,CAT和LOX活性逐渐升高,而POD活性呈先降低后升高的变化趋势。脱水后,种子中MDA含量均高于CK。60DAF的沙芥种子已获得脱水耐性。     

Biochemical Characteristics Associated with the Development of the Desiccation-sensitive Seeds of Machilus thunbergii Sieb. & Zucc.

Biochemical properties, i.e. endogenous abscisic acid, proline,sugars, respiration, adenosine phosphates and adenylate energycharge, and growth and moisture content were measured duringthe development of seeds of Machilus thunbergii. As dry matteraccumulated in the embryo during development, moisture content,ABA, proline, respiration and sugars all declined. At maturity,the dry mass of the seeds failed to attain a plateau beforethe period of natural seed shedding; the axis and cotyledonsreached moisture contents of 58 and 45%, respectively. Dryingof immature seeds at 73% relative humidity and 25 °C for30 d resulted in a complete loss of viability at all developmentalstages tested with the exception of mature seeds that were ableto tolerate a 5% decrease in moisture content before germinationdeclined. ABA was detected in all embryos tested, with a maximum value16.·16 µg g^-1 d. wt about midway through development.Although the presence of ABA induced no tolerance to desiccationof mature seeds, it did coincide with decreased content of waterin the developing seeds and decreased respiration. Desiccationdamage of M. thunbergii seeds occurred when moisture contentwas still high (45%) and this damage was not related to theabsence of oligosaccharides in the mature seeds. We concludethat developing embryos and mature seeds of M. thunbergii haveproperties common to many recalcitrant seeds, with seeds beingsensitive to desiccation at all stages, having a prominent ABApeak, little proline, lacking oligosaccharides, and specifically,little dormancy and a moderate rate of respiration of matureseeds (0·9 µmol O₂ min^-1 g^-1 f. wt). Adenosinetriphosphate content and energy charge decreased from stagefour to stage eight of seed development, then increased againto 103 nmol g^-1 d. wt and 0·73, respectively, in matureseeds. The moderate energy charge observed in mature seeds indicatesthat continuous metabolism is also a characteristic of recalcitrantseeds.Copyright 1995, 1999 Academic Press Machilus thunbergii, seed development, recalcitrant seed, abscisic acid, energy charge     

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.     

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     

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     

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。     

Different quiescence states of three culture cell lines detected by acridine orange staining of cellular RNA

Three cell lines (CHO, L-929, and R1H) were investigated for their growth kinetics and the difference of exponential and quiescent state of monolayers in medium with and without serum (L-929). The noncycling populations of L-929 and R1H in medium with serum contained increased G1-phase percentages but also considerable proportions of SQ and G2Q cells. Although about 90% of the cells excluded trypan blue, the viability tested by colony assay was clearly lower than for exponentially growing cultures. CHO cells showed similar fractions of cells in G1-, S-, and G2-Q compartments but in addition considerable cell loss. The RNA content of these cells was reduced in plateau phase by 7-48% depending on cell type and residence time in the noncycling state. The data suggest that the cells suffered from nutrition depletion and were arrested in all phases of the cycle. In contrast, L-929 cells in medium without serum reduced their RNA content down to one-third that of proliferating cells and still retained the full viability as shown by the same plating efficiency in a colony assay. Since about 90% of the cells had G1 DNA content, these cells resemble true G1Q or G0 cells controlled by growth factors rather than nutritional depletion.     

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.