Abscisic acid and osmoticum prevent germination of developing alfalfa embryos,but only osmoticum maintains the synthesis of developmental proteins

Developing seeds of alfalfa (Medicago sativa L.) acquire the ability to germinate during the latter stages of development, the maturation drying phase. Isolated embryos placed on Murashige and Skoog medium germinate well during early and late development, but poorly during mid-development; however, when placed on water they germinate well only during the latter stage of development. Germination of isolated embryos is very slow and poor when they are incubated in the presence of surrounding seed structures (the endosperm or seed coat) taken from the mid-development stages. This inhibitory effect is also achieved by incubating embryos in 10^?5 M abscisic acid (ABA). Endogenous ABA attains a high level during mid-development, especially in the endosperm. Seeds developing in pods treated with fluridone (1-methyl-3-phenyl-5[3-(trifluoromethyl)-phenyl]-4(1H)-pyridinone) contain low levels of ABA during mid-development, and the endosperm and seed coat only weakly inhibit the germination of isolated embryos. However, intact seeds from fluridone-treated pods do not germinate viviparously, which is indicative that ABA alone is not responsible for maintaining seeds in a developing state. Application of osmoticum (e.g. 0.35 M sucrose) to isolated developing embryos prevents their germination. Also, in the developing seed in situ the osmotic potential is high. Thus internal levels of osmoticum may play a role in preventing germination of the embryo and maintaining development. Abscisic acid and osmoticum impart distinctly different metabolic responses on developing embryos, as demonstrated by their protein-synthetic capacity. Only in the presence of osmoticum do embryos synthesize proteins which are distinctly recognizable as those synthesized by developing embryos in situ, i.e. when inside the pod. Abscisic acid induces the synthesis of a few unique proteins, but these arise even in mature embryos treated with ABA. Thus while both osmoticum and ABA prevent precocious germination, their effects on the synthetic capacity of the developing embryo are quite distinct. Since seeds with low endogenous ABA do not germinate, osmotic regulation may be the more important of these two factors in controlling seed development.     

Seeds of tomato (Lycopersicon esculentum Mill.) which develop in a fully hydrated environment in the fruit switch from a developmental to a germinative mode without a requirement for desiccation

Seed water content is high during early development of tomato seeds (10–30 d after pollination (DAP)), declines at 35 DAP, then increases slightly during fruit ripening (following 50 DAP). The seed does not undergo maturation drying. Protein content during seed development peaks at 35 DAP in the embryo, while in the endosperm it exhibits a triphasic accumulation pattern. Peaks in endosperm protein deposition correspond to changes in endosperm morphology (i.e. formation of the hard endosperm) and are largely the consequence of increases in storage proteins. Storage-protein deposition commences at 20 DAP in the embryo and endosperm; both tissues accumulate identical proteins. Embryo maturation is complete by 40 DAP, when maximum embryo protein content, size and seed dry weight are attained. Seeds are tolerant of premature drying (fast and slow drying) from 40 DAP.Thirty-and 35-DAP seeds when removed from the fruit tissue and imbibed on water, complete germination by 120 h after isolation. Only seeds which have developed to 35 DAP produce viable seedlings. The inability of isolated 30-DAP seed to form viable seedlings appears to be related to a lack of stored nutrients, since the germinability of excised embryos (20 DAP and onwards) placed on Murashige and Skoog (1962, Physiol. Plant. 15, 473–497) medium is high. The switch from a developmental to germinative mode in the excised 30- and 35-DAP imbibed seeds is reflected in the pattern of in-vivo protein synthesis. Developmental and germinative proteins are present in the embryo and endosperm of the 30- and 35-DAP seeds 12 h after their isolation from the fruit. The mature seed (60 DAP) exhibits germinative protein synthesis from the earliest time of imbibition. The fruit environment prevents precocious germination of developing seeds, since the switch from development to germination requires only their removal from the fruit tissue.Abbreviations DAP days after pollination - kDa kilodaltons - SP1-4 storage proteins 1–4 - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis - HASI hours after seed isolation - MS medium Murashige and Skoog (1962) medium This work is supported by National Science and Engineering Research Council of Canada grant A2210 to J.D.B.     

The Role of Maturation Drying in the Transition from Seed Development to Germination: VII. EFFECTS OF PARTIAL AND COMPLETE DESICCATION ON ABSCISIC ACID LEVELS AND SENSITIVITY IN RICINUS COMMUNIS L. SEEDS

During mid-development (25–40 d after pollination: DAP)of the castor bean seed the amount of abscisic acid (ABA) increasesin both the endosperm and the embryo, declining substantiallythereafter until there is little present in the mature dry (60DAP) seed. Premature desiccation of the seed at 35 DAP alsoleads to a major decline in ABA within the embryo and endosperm.Partial water loss from the seed at 35 DAP which, like naturaland premature desiccation, leads to subsequent germination uponreturn of the seed to full hydration, causes a much smallerdecline in ABA levels. In contrast, ABA declines substantiallyin the non-dried (hydrated) control at 35 DAP, but the seedsdo not germinate. Hence, a clear negative correlation betweenABA content and germinability is not observed. Both drying,whether natural or imposed prematurely, and partial drying decreasethe sensitivity of the isolated embryo to exogenous ABA by about10-fold. The protein synthetic response of the castor bean embryo exposedto 0.1 mol m^–3 ABA following premature desiccation exhibitssome similarity to the response of the non-dried developingembryo—in both cases the synthesis of some developmentalproteins is enhanced by ABA, and germination is suppressed.Germination of mature seeds is also suppressed by 0.1 mol m^–3ABA, but the same developmental proteins are not synthesized.In the cotyledons of prematurely-desiccated seed, some proteinsare hydrolysed upon imbibition in 0.1 mol m^–3 ABA, a phenomenonthat occurs also in the cotyledons of similarly treated matureembryos, but not in developing non-dried embryos. Hence theembryo exhibits an ‘intermediate’ response uponrehydration in 0.1 mol m^–3 ABA following premature desiccation;viz. some of the responses are developmental and some germinative.Following natural or imposed drying, the isolated embryo becomesrelatively insensitive to 0.01 mol m^–3 ABA: germinationis elicited and post-germinative reserve breakdown occurs inthe radicle and cotyledons. The reduced sensitivity of the embryoto ABA as a consequence of desiccation may be an important factorin eliciting the switch to germination and growth within thewhole seed. Key words: Abscisic acid, desiccation, astor bean endosperm, seed development, germination, protein synthesis, isolated embryos, hormone sensitivity     

Effect of Water Stress, Seed Coat Restraint, and Abscisic Acid upon Different Germination Capabilities of Two Tomato Lines at Low Temperature

Two tomato (Lycopersicon esculentum Mill.) lines with greatly different capabilities to germinate at 10°C were compared with respect to sensitivity to experimental treatments which affect the water status of the embryo. Germination rates and final percentages could be drastically changed (at 25°C) by (a) removing the mechanical constraint from the radicle tip, (b) imposing water stress by an osmoticum, (c) stress hardening of the seeds through osmotic pretreatment, and (d) inhibiting embryo expansion by abscisic acid (ABA). All treatments showed a similar difference in germination vigor between the two lines indicating that cold sensitivity is in fact a matter of water relations rather than of phase transitions in membrane lipids. Inhibition of germination by ABA was completely abolished by removing the mechanical constraint from the radicle tip. Osmotic stress of −3 bar which quantitatively replaced this constraint in inhibiting germination also restored the sensitivity to ABA. It is concluded that all these treatments act on the balance between the hydraulic extension force of the embryo radicle and the opposing force of the seed layers covering the radicle tip. The difference in cold sensitivity between the two seed lines resides either in the osmotic potential or the pressure potential of the germinating embryo.     

Germination and Dormancy of Abscisic Acid- and Gibberellin-Deficient Mutant Tomato (Lycopersicon esculentum) Seeds (Sensitivity of Germination to Abscisic Acid,Gibberellin, and Water Potential)

Germination responses of wild-type (MM), abscisic acid (ABA)-deficient (sitw), and gibberellin (GA)-deficient (gib-1) mutant tomato (Lycopersicon esculentum Mill. cv Moneymaker) seeds to ABA, GA4+7, reduced water potential ([psi]), and their combinations were analyzed using a population-based threshold model (B.R. Ni and K.J. Bradford [1992] Plant Physiol 98: 1057-1068). Among the three genotypes, sitw seeds germinated rapidly and completely in water, MM seeds germinated more slowly and were partially dormant, and gib-1 seeds did not germinate without exogenous GA4+7. Times to germination were inversely proportional to the differences between the external osmoticum, ABA, or GA4+7 concentrations and the corresponding threshold levels that would either prevent ([psi]b, log[ABAb]) or promote (log[GAb]) germination. The sensitivity of germination to ABA, GA4+7, and [psi] varied widely among individual seeds in the population, resulting in a distribution of germination times. The rapid germination rate of sitw seeds was attributable to their low mean [psi]b (-1.17 MPa). Postharvest dormancy in MM seeds was due to a high mean [psi]b (-0.35 MPa) and a distribution of [psi]b among seeds such that some seeds were unable to germinate even on water. GA4+7 (100 [mu]M) stimulated germination of MM and gib-1 seeds by lowering the mean [psi]b to -0.75 MPa, whereas ABA inhibited germination of MM and sitw seeds by increasing the mean [psi]b. The changes in [psi]b were not due to changes in embryo osmotic potential. Rather, hormonal effects on endosperm weakening opposite the radicle tip apparently determine the threshold [psi] for germination. The analysis demonstrates that ABA- and GA-dependent changes in seed dormancy and germination rates, whether due to endogenous or exogenous growth regulators, are based primarily upon corresponding shifts in the [psi] thresholds for radicle emergence. The [psi] thresholds, in turn, determine both the rate and final extent of germination within the seed population.     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.) : III. Sensitivity of Germination to Water Potential and Abscisic Acid during Development

Muskmelon (Cucumis melo L.) seeds are germinable 15 to 20 days before fruit maturity and are held at relatively high water content within the fruit, yet little precocious germination is observed. To investigate two possible factors preventing precocious germination, the inhibitory effects of abscisic acid and osmoticum on muskmelon seed germination were determined throughout development. Seeds were harvested at 5-day intervals from 30 to 65 days after anthesis (DAA) and incubated either fresh or after drying on factorial combinations of 0, 1, 3.3, 10, or 33 micromolar abscisic acid (ABA) and 0, −0.2, −0.4, −0.6, or −0.8 megapascals polyethylene glycol 8000 solutions at 30°C. Radicle emergence was scored at 12-hour intervals for 10 days. In the absence of ABA, the water potential (Ψ) required to inhibit fresh seed germination by 50% decreased from −0.3 to −0.8 megapascals between 30 and 60 DAA. The Ψ inside developing fruits was from 0.4 to 1.4 megapascals lower than that required for germination at all stages of development, indicating that the fruit Ψ is sufficiently low to prevent precocious germination. At 0 megapascal, the ABA concentration required to inhibit germination by 50% was approximately 10 micromolar up to 50 DAA and increased to >33 micromolar thereafter. Dehydration improved subsequent germination of immature seeds in ABA or low Ψ. There was a linear additive interaction between ABA and Ψ such that 10 micromolar ABA or −0.5 megapascal osmotic potential resulted in equivalent, and additive, reductions in germination rate and percentage of mature seeds. Abscisic acid had no effect on embryo solute potential or water content, but increased the apparent minimum turgor required for germination. ABA and osmoticum appear to influence germination rates and percentages by reducing the embryo growth potential (turgor in excess of a minimum threshold turgor) but via different mechanisms. Abscisic acid apparently increases the minimum turgor threshold, while low Ψ reduces turgor by reducing seed water content.     

Dormancy and Germination of Abscisic Acid-Deficient Tomato Seeds : Studies with the sitiens Mutant

The role of abscisic acid (ABA) in the dormancy induction of tomato (Lycopersicon esculentum) seeds was studied by comparison of the germination behavior of the ABA-deficient sitiens mutant with that of the isogenic wild-type genotype. Freshly harvested mutant seeds, in contrast to wild-type seeds, always readily germinate and even exhibit viviparous germination in overripe fruits. Crosses between mutant and wild-type and self-pollination of heterozygous plants show that in particular the ABA fraction of embryo and endosperm is decisive for the induction of dormancy. After-ripened wild-type seeds fully germinate in water but are more sensitive toward osmotic inhibition than mutant seeds. Germination of both wild-type and mutant seeds is equally sensitive toward inhibition by exogenous ABA. ABA content of mature wild-type seeds is about 10-fold the level found in mutant seeds. Nevertheless, it is argued that the differences in dormancy between the seeds of both genotypes are not a result of actual ABA levels in the mature seeds or fruits but a result of differences in ABA levels during seed development. It is hypothesized that the high levels of ABA that occur during seed development in wild-type seeds induce an inhibition of cell elongation of the radicle that can still be observed after long periods of dry storage.     

The Role of Maturation Drying in the Transition from Seed Development to Germination: V. RESPONSES OF THE IMMATURE CASTOR BEAN EMBRYO TO ISOLATION FROM THE WHOLE SEED: A COMPARISON WITH PREMATURE DESICCATION

Kennode, A. R, and Bewley, J. D. 1988. The role of maturationdrying in the transition from seed development to germination.V. Responses of the immature castor bean embryo to isolationfrom the whole seed; a comparison with premature desiccation.—J.exp. Bot. 39: 487–497. Desiccation is an absolute requirement for germination and post-germinativegrowth of whole seeds of the castor bean, whether desiccationis imposed prematurely during development, at 35 d after pollination(DAP) or occurs naturally during late maturation (50–60DAP). Desiccation also plays a direct role in the inductionof post-germinative enzyme synthesis in the cotyledons of embryosin the intact seed; this event is not simply due to the presenceof a growing axis. Isolation of embryos from the developingcastor bean seed at 35 DAP results in both germination and growth,despite the absence of a desiccation event. We have comparedthe metabolic consequences of premature drying of whole seeds(35 DAP) and isolation of the developing 35 DAP embryos. Inboth cases, hydrolytic events involved in the mobilization ofstored protein reserves proceed in a similar manner and mirrorthose events occurring within germinated mature seeds. Thereare differences, however, for post-germinative enzyme (LeuNAaseand isocitrate lyase) production occurs to a lesser extent innon-dried isolated embryos than in those from prematurely dried(35 DAP) whole seeds, or from mature dry (whole) seeds. Desiccationof the 35 DAP whole seed does not alter the subsequent responseof the embryo upon isolation. Thus, while drying does not affectthe metabolism of isolated embryos, it has a profound effecton that of embryos within the intact seed. Tissues surroundingthe embryo in the developing intact seed (viz. the endosperm)maintain its metabolism in a developmental mode and inhibitgermination. This effect of the surrounding tissues can onlybe overcome by drying or by their removal. Key words: Metabolism, isolation, desiccation, embryo, endosperm, castor bean, development, germination     

Endogenous abscisic acid during the germination of chick-pea seeds

Over the past twenty years many studies have been undertaken to elucidate the regulation of seed germination. Abscisic acid (ABA) and the gibberellins (GAs) are the hormones proposed to control this process, the first by inhibiting and the second by inducing germination. It has been proposed that a high water potential increases the growth potential of the embryo, presumably permitting the production or activation by GA of the cell wall hydrolases and thus decreasing the yield threshold of the endosperm close to the radicle tip. A low water potential, e.g., imbibition in an osmoticum. imposes a stress on cell metabolism, by reducing the turgor of the radicle cells, and there is a decrease in growth potential. Exogenous ABA also causes a decline in growth potential of the radicle: however, the actions of low water potential in preventing germination are not mediated through an increase in ABA in the seeds. In the present paper an attempt is made to asses the role of ABA and polyethylene glycol (PEG) in the germination of chick-pea (Cicer arietinum L.) seeds. The endogenous ABA of chick-pea seeds was purified by reversed-phase HPLC and quantified by GC-ECD. The variations in the ABA levels in the embryonic axes and the cotyledons were studied during 120 h. of imbibition. The highest ABA level in the embryome axes was found at 18 h. coinciding with an increase in fresh weight and a high germination percentage. ABA was not detected in the cotyledons during incubation which probably indicates that the hormone is more involved in the active growth of the embryonic axes itself than in the mobilization process of the reserves. When seeds were treated with different PEG-cycles. PEG delayed germination, reduced the fresh weight of embryonic axes, and retarded the onset of ABA synthesis. It is concluded that endogenous ABA is related to the onset of germination and the growth of the embryonic axis. In addition, there is no correlation among the different PEG-cycles and the level of ABA and germination. Germination was related more to the water conditions inside the embryo's cells than to ABA levels.     

内源脱落酸和赤霉素对番茄发育果实和种子水分关系的影响

利用热偶湿度计（ｔｈｅｒｍｏｃｏｕｐｌｅｐｓｙｃｈｒｏｍｅｔｅｒ）研究了野生型、ＧＡ－缺陷型和ＡＢＡ－缺陷型番茄发育过程中果实种子的水分关系,发现除ＡＢＡ－缺陷型种子胶囊和果肉水势变化特殊外,３种类型果实水分状况变化基本一致;在整个发育时期内．前期种子胶囊和果肉水分流向种子,中期种子水分流向种子胶囊和果肉,后期种子和果实间的水势达到平衡。鉴于种胚脱水是一种主动过程,种胚水势一直低于整个种子、种子胶囊和果肉。内源赤霉素可明显增加果实和种子的重量,但对增加种胚溶质的作用不大。由于内源脱落酸可以促使果实成熟和衰老,促进果实细胞解体,大大降低种子胶囊和果肉水势,因而抑制成熟种子在果实内萌发。     

Embryo and hip development in hybrid roses

Roses are known to produce seeds with high concentrations of abscisic acid (ABA), both in the pericarp and in the testa tissues of the seed coat. No studies on roses have documented embryo morphological differentiation or the concentration of ABA in the embryo, which is known to inhibit premature germination. In this study, hip and seed growth of two hybrid roses were characterised from 3 to 60 days after pollination (DAP). An increase of about five times the hip mass at 3 DAP was necessary to obtain fully developed seeds. Fully developed embryos were found at 15 DAP and completely developed seeds at 30 DAP. The same pattern of hip mass increase was shown in both genotypes. In parallel, quantification of ABA in the developing embryos was carried out by ELISA. An exponential decay in ABA concentration was found in embryos of both genotypes, with basal levels (<0.5 pmol mg^?1) registered at 30 DAP. These changes in ABA, during the embryo development, could be used to formulate time points for embryo rescue and understanding of the pollination to seedling stage.     

Storage behavior of Chionanthus retusus seed and asynchronous development of the radicle and shoot apex during germination in relation to germination inhibitors, including abscisic acid and four phenolic glucosides

Studies on seed storage of Chionanthus retusus Lindl. & Paxt. revealed an orthodox behavior, one which showed both desiccation and freezing tolerance. An epicotyl after-ripening dormancy was expressed in C. retusus seeds by slow growth of the shoot apex relative to more rapid growth of the radicle when seeds were germinated at 30/20 degrees C. Although these seeds exhibit radicle protrusion, they must be after-ripened for another 8-10 weeks at 30/20 degrees C in order to obtain normal shoot growth. Removal of the endosperm, however, quickly stimulated cotyledon and shoot emergence without the additional after-ripening. Water-soluble glucoside phenolics, GL-3, Nuzhenide, ligustroside and oleoside dimethyl ester are present at relatively high levels in endosperm of freshly harvested seeds. These glucoside phenolics are excreted from the endosperm during subsequent after-ripening. Embryo and endosperm tissue from seed germinating at 30/20 degrees C (germination being defined by protrusion of the radicle) had a 10 times lower abscisic acid (ABA) content than similar tissues from freshly harvested mature seed. However, no shoot growth occurred even with the 10-fold reduction in ABA and a concomitant increase in endogenous gibberellins A1, A4 and A20. Thus, epicotyl dormancy during the first 8 weeks of after-ripening at 30/20 degrees C may be controlled by factors other than high ABA, i.e., the slow development of the shoot apex following radicle protrusion may be controlled more by high levels of glucoside phenolics than by diminished ABA and elevated GA levels.     

一种刺激肉苁蓉种子萌发和吸器发育的方法

以3种肉苁蓉属（Cistanche）植物种子为材料,研究了氟啶酮对肉苁蓉种子萌发和吸器发育的影响。结果表明：氟啶酮处理10天后种子开始萌发,然后,胚根顶端膨大,发育出吸器。管花肉苁蓉种子萌发率最高,达到82％;盐生肉苁蓉吸器诱导率最高,达到52％;对照处理的种子不萌发。这说明氟啶酮是有效刺激肉苁蓉种子萌发和吸器发育的信号物质。     

Involvement of ABA in induction of secondary dormancy in barley (Hordeum vulgare L.) seeds

At harvest, barley seeds are dormant because their germination is difficult above 20 degrees C. Incubation of primary dormant seeds at 30 degrees C, a temperature at which they do not germinate, results in a loss of their ability to germinate at 20 degrees C. This phenomenon which corresponds to an induction of a secondary dormancy is already observed after a pre-treatment at 30 degrees C as short as 4-6 h, and is optimal after 24-48 h. It is associated with maintenance of a high level of embryo ABA content during seed incubation at 30 degrees C, and after seed transfer at 20 degrees C, while ABA content decreases rapidly in embryos of primary dormant seeds placed directly at 20 degrees C. Induction of secondary dormancy also results in an increase in embryo responsiveness to ABA at 20 degrees C. Application of ABA during seed treatment at 30 degrees C has no significant additive effect on the further germination at 20 degrees C. In contrast, incubation of primary dormant seeds at 20 degrees C for 48 and 72 h in the presence of ABA inhibits further germination on water similarly to 24-48 h incubation at 30 degrees C. However fluridone, an inhibitor of ABA synthesis, applied during incubation of the grains at 30 degrees C has only a slight effect on ABA content and secondary dormancy. Expression of genes involved in ABA metabolism (HvABA8'OH-1, HvNCED1 and HvNCED2) was studied in relation to the expression of primary and secondary dormancies. The results presented suggest a specific role for HvNCED1 and HvNCED2 in regulation of ABA synthesis in secondary seed dormancy.     

一种刺激肉苁蓉种子萌发和吸器发育的方法

以3种肉苁蓉属(Cistanche)植物种子为材料, 研究了氟啶酮对肉苁蓉种子萌发和吸器发育的影响。结果表明: 氟啶酮处理10天后种子开始萌发, 然后, 胚根顶端膨大, 发育出吸器。管花肉苁蓉种子萌发率最高, 达到82%; 盐生肉苁蓉吸器诱导率最高, 达到52%; 对照处理的种子不萌发。这说明氟啶酮是有效刺激肉苁蓉种子萌发和吸器发育的信号物质。     

Expression of an expansin is associated with endosperm weakening during tomato seed germination

Expansins are extracellular proteins that facilitate cell wall extension, possibly by disrupting hydrogen bonding between hemicellulosic wall components and cellulose microfibrils. In addition, some expansins are expressed in non-growing tissues such as ripening fruits, where they may contribute to cell wall disassembly associated with tissue softening. We have identified at least three expansin genes that are expressed in tomato (Lycopersicon esculentum Mill.) seeds during germination. Among these, LeEXP4 mRNA is specifically localized to the micropylar endosperm cap region, suggesting that the protein might contribute to tissue weakening that is required for radicle emergence. In gibberellin (GA)-deficient (gib-1) mutant seeds, which germinate only in the presence of exogenous GA, GA induces the expression of LeEXP4 within 12 hours of imbibition. When gib-1 seeds were imbibed in GA solution combined with 100 microM abscisic acid, the expression of LeEXP4 was not reduced, although radicle emergence was inhibited. In wild-type seeds, LeEXP4 mRNA accumulation was blocked by far-red light and decreased by low water potential but was not affected by abscisic acid. The presence of LeEXP4 mRNA during seed germination parallels endosperm cap weakening determined by puncture force analysis. We hypothesize that LeEXP4 is involved in the regulation of seed germination by contributing to cell wall disassembly associated with endosperm cap weakening.     

Secondary dormancy (skotodormancy) in seeds of lettuce (Lactuca sativa cv. Grand Rapids) and its release by light, gibberellic acid and benzyladenine

Lettuce seeds (Lactuca sativa L. cv. Grand Rapids) imbibed in darkness at supra-optimal temperatures (23 ± 1°C) develop a secondary dormancy, termed skotodormancy. The seeds first lose their ability to be promoted to germinate by gibberellic acid, and then lose their ability to be promoted by red light. A combination of red light and gibberellic acid will break skotodormancy for longer than either alone, but red light and benzyladenine together are much more effective. Desiccation of skotodormant seeds does not diminish their dormancy. Embryos dissected from skotodormant seeds will germinate, and are as capable of radicle expansion in the osmoticum polyethylene glycol as are newly-imbibed seeds. Hence skotodormancy is a whole seed dormancy and does not reside within the embryo as an inherent block to germination processes, but as an inability to respond to the stimulation of red light or to hormone.     

The effect of excess moisture on the germination of Spinacia oleracea L.

Summary The reversible inhibition of the germination of spinach (Spinacia oleracea L.) seeds in conditions which are even slightly wetter than optimal has been traced to the production, in a wet environment, of a layer of mucilage around and within the fruit coat which surrounds the true seed. Such wet seeds may however germinate readily when the temperature is lowered, or the oxygen pressure of the environment is raised, or the intact seeds are placed for a short time in hydrogen peroxide before being transferred to what normally would be an excess of water. Even in the absence of an increased oxygen supply the seeds will germinate under water provided the fruit coat, or even a small part of it where it covers the radicle, is crefully removed. No evidence has been found of a water soluble inhibitor and the findings are consistent with the hypothesis that germination is dependent on a sufficiently high rate of supply of oxygen to the sites embryonic respiration. The mucilage which is formed under wet conditions forms a barrier which prevents the transfer of oxygen to the embryo by gaseous diffusion or aqueous convection currents and restricts it to the process of aqueous diffusion, and under these conditions the rate of oxygen supply may not reach the threshold level required for germination.     

香圆种子萌发特征及催芽技术

以香圆（Citrus wilsonii Tanaka）种子为实验材料,通过测定种皮透水性、种皮机械障碍、发芽温度等对种子萌发的影响从而探讨种子萌发特征;通过切割种子、热水浸种、赤霉素浸种、低温层积等处理方式探索种子的催芽方法。结果显示：香圆种皮不存在透水性障碍,但其机械障碍对种子萌发具有抑制作用,且内种皮对萌发影响较大;种子在15℃、25℃、35℃恒温及15℃/30℃变温条件下均可萌发,其最适萌发温度为25℃;采用低温层积、热水浸种、切除1/3种子等方法均可一定程度上促进种子萌发。生产中宜采用始温60℃热水浸种24h的方法对香圆种子进行催芽。