Dormancy in Seeds of Charlock: II. THE INFLUENCE OF THE SEED COAT

An attempt was made to elucidate those factors associated withthe seed coat which are responsible for maintaining the dormancyof the charlock seed. The rates of water uptake by the seedand by the embryo were measured; the permeability of the coveringlayers to water was much the same as that of parenchymatouscells. The action of certain dormancy-breaking treatments, viz.embryo excision, exposure to concentrated sulphuric acid, alternatingtemperatures, and gibberellic acid, were therefore examined.Fromexperiments with excised embryos and seeds exposed to concentratedsulphuric acid, it appeared that the loss of dormancy followingthe removal of the seed coat resulted from an increase in oxygensupply to the embryo. The germination of a small proportionof seeds at low temperature may also be due to an increase inthe oxygen concentration within the seed. The dormancy-breakingeffect of gibberellic acid, however, is unlikely to be due tochanges in the permeability of the seed coat to oxygen. Estimatesof the rate of oxygen uptake and growth of seeds treated withgibberellic acid at various external oxygen concentrations showthat the resistance of the covering layers to diffusion of oxygenretards, but does not prevent, the initiation of growth.It wouldappear that dormancy is maintained in charlock seeds by theaction of a specific growth-inhibiting substance which is producedat low oxygen concentration in the interior of the seed andwhich diffuses to the meristems.     

Models of Oxygen Diffusion in Respiring Seed

Models of oxygen diffusion in respiring seeds of uniform propertiesand with a relatively impermeable seed coat are proposed. Specimencalculations are presented using the data of Edwards (1969)for excised charlock embryos and intact seeds. The calculatedanswers are very sensitive to the assumed values of the oxygendiffusion coefficient and absorption coefficient in the seedmaterial but whatever the assumptions, the apparent dormancyof charlock seed in air may be directly related to an oxygendeficiency within the embryo tissue. The consequences of errorsin measured values of the diffusion and absorption coefficientsin the seed material, and of assumptions concerning the uniformityof seed material, are discussed. Since the ultimate test of a diffusion model depends upon anaccurate measurement of the concentration distribution withinthe seed, the useful limit of development of respiration andoxygen diffusion models may have been reached because of thepractical difficulties in making these measurements.     

水分胁迫条件下高粱种子的吸水机制及其可溶性糖代谢的变化

以高梁杂交种晋杂８６－１为材料研究了种子吸水萌动过程中胚与胚乳两部分的水分状况及可溶性糖变化的过程。结果表明：在吸水萌动过程中种胚和胚乳在水分及糖代谢方面具有不同的特点。     

Dormancy in Seeds of Charlock: I. DEVELOPMENTAL ANATOMY OF THE SEED

The anatomy of the embryo, endosperm, and testa throughout thedevelopment of charlock seeds is described. In mature seedsthe embryos are morphologically fully differentiated. Each embryoconsists of a shoot meristem with two large cotyledons but noleaf primordia, and a root meristem with a root cap. The embryois surrounded by a single layer of aleurone-like cells derivedfrom the endosperm and enclosed within the testa. Mucilagesand phenols which could greatly retard the diffusion of oxygeninto the tissues of the embryo are found in the testa. Dormancy,which appears to be induced by a shortage of oxygen, is probablyassociated with the presence of these covering layers.     

刺楸种子中性与酸性抑制物质的研究

本文研究了刺楸种子抑制物质的提取,分离和鉴定方法,并测定了抑制物质的相对活性及其含量。结果表明,刺揪种子中除含高水平的酸性抑制物外,还存在一种中性抑制物。该中性抑制物同酸性抑制物相似,有较强的生物活性,不但可以抑制白菜种子的萌发与胚根生长,而且对已解除休眠的刺楸种子也有显著作用。利用纸层析,薄层层析,颜色反应和高效液相色谱等手段,证明中性抑制物质为香豆素类物质;酸性抑制物质的主要成分为脱落酸。种子各部位均含这两种抑制物质,但含量有所不同,随种子贮藏时间的延长,抑制物会发生转移。本文还讨论了用不同溶剂提取中性抑制物的效果。     

Photoinhibition of radish (Raphanus sativus L) seed germination: control of growth potential by cell-wall yielding in the embryo

The biophysical mechanism underlying photoinhibition of radish (Raphanus sativus L.) seed germination was investigated using three cultivars differing in sensitivity to continuous irradiation with far-red light (high-irradiance reaction of phytochrome). Sensitivity of germination to the inhibitory action of light was assessed by probing germination under osmotic stress (incubation in media of low water potentials adjusted with polyethylene glycol 6000) and expressed in terms of ‘germination potential’ (positive value of the water potential at which germination is inhibited by 50%). Far-red light decreases the germination potential to various degrees in the different cultivars, reflecting the light-sensitivity of germination in water. Removal of the seed coat increases the germination potential by a constant amount in darkness and light. It is concluded that germination depends on the expansive force of the embryo which can be drastically diminished by far-red light. Seed-coat constraint and expansive force of the embryo interact additively on the level of the germination potential. Photoinhibition of germination was accompanied by an inhibition of water uptake into the seed. Analysis of seed water relations showed that osmotic pressure and turgor assumed higher levels in photoinhibited seeds, compared to seeds germinating in darkness, while the water potential was close to zero under both conditions. Far-red light produced a shift (to less negative values) in the curve relating water-uptake rate to external water potential, i.e. a reduction in the driving force for water uptake. It is concluded that photoinhibition of germination results from the maintenance of a high threshold of cell-wall extensibility in the embryo.     

Water Relations of Tomato Seeds During Imbibition and Osmotic Priming

Water uptake of tomato (Lycopersicon esculentum Mill. cv. Moneymaker) seeds during germination was obviously triphasic. The completion of the first phase of water uptake by whole seed could not be realized until 10~12 h later after sowing though varies in different parts of seed. The mechanical resistance of endosperm and seed coat restricted water uptake of the embryo envoleped by the endosperm. Water potential of the intact embryo was still 0. 6~0. 9 Mpa lower than the whole seed when the equilibrium between seed and imbibing solution was established. GA and ABA had no direct effects on the water uptake of tomato seeds. The water potential of embryo was positively correlated with its moisture content. The osmotic potential of tomato embryos decreased slowly during imbibition in water and osmotic solution as well.     

Lucerne Saponins as Inhibitors of Cotton Seed Germination: THEIR EFFECT ON DIFFUSION OF OXYGEN THROUGH THE SEED COATS

The site of inhibition of cotton-seed germination by lucernesaponins seems to be associated with tissues other than theembryo. The rate of water uptake during imbibition was similarfor intact seeds immersed in water and for those in 0.5 percent saponin solution. Seeds previously immersed in saponinsolution showed a lower rate of respiration, absorbing about60 per cent less oxygen than those pre-immersed in water. Yetthe respiration rate of excised embryos from both treatmentswas the same. The extent of the inhibition of germination wasnot affected when oxygen was either bubbled through the saponin-containingimmersion-solution or passed through the flasks in which theseeds were germinated after 24 h of pre-immersion. The rates of oxygen diffusion through the seed coats and membranesof seeds pre-immersed in water or in saponin solution were measured.They were distinctly lower for seed coats and membranes whichhad been exposed to the saponin treatment. It has been concludedthat the seed coat and/or the membrane of intact cotton seedspre-immersed in saponin solution serve as a barrier for oxygendiffusion to the embryo, resulting in inhibition of germination.     

Water Relations of Beetroot Seed Germination II. Effects of the Ovary Cap and of the Endogenous Inhibitors

Conditions in which seeds of beetroot (Beta vulgaris L.) willgerminate are relatively narrowly limited by a deficient oran excessive water-supply. The ovary cap, which covers eachseed situated within its locule in the seed cluster, has beenshown to be responsible, under wet conditions, for preventingaccess of oxygen to the embryo, owing to its imperviousnessto gas and to the production of mucilage around its rim in thepresence of excess water. Seeds in intact clusters germinatein the wet if the oxygen pressure is increased. Removal of theovary cap enables seeds to germinate even under water. Preliminarywashing of the clusters increases germination under dry conditions,owing to the elimination from them of an endogenous water-solublegermination inhibitor complex. Washing also improves germinationunder wet conditions; and measurements of oxygen uptake suggestthat the dilute inhibitor may further depress the respirationalready obstructed by the ovary cap. Conversely, removal ofthe ovary cap appears to have the same effect as eliminatingthe inhibitor complex. Depressed germination is usually correlatedwith depressed oxygen uptake of the imbibing seeds but an exceptionallyhigh concentration of inhibitor can uncouple respiration. The production of beetroot seeds with a low level of inhibitorin the cluster material and with loose and non-mucilaginousovary caps, or the washing and drying of clusters prior to sowingshould widen the range of moisture conditions over which beetrootseeds are able to germinate.     

芡实种子萌发期的生物学特性与结构解剖

本文描述了芡实种子的结构,种子的萌发和幼苗的形态特征。成熟种子必须置于水中保存,以增大胚体,并完成后熟作用。外胚乳是种子萌发和幼苗生长的主要营养来源。萌发后在子叶叶柄基部外侧形成的突起结构可能起固着作用。     

Is The Barley Endosperm a Water Reservoir for the Embryo When Germinating Seeds Are Dried?

The water content of germinating seeds fluctuates in response to water potential changes in the surrounding environment. We tested the hypothesis that the endosperm functions as a water reservoir when imbibed seeds experience drying, and we characterized water uptake and movement within barley (Hordeum vulgare cv. Triumph) caryopses (hereafter referred to as seeds). Water movement into and through germinating barley seeds during imbibition and drying was determined gravimetrically and with the fluorescent dye trisodium 8-hydroxy-1,3,6-pyrenetrisulfonate (PTS). During imbibition, embryo tissues hydrated more rapidly and reached a higher water content (g H20/g dry weight) than did the endosperm, although the endosperm eventually contained nine times as much total water. When barley seeds that had imbibed for 12 h were exposed to moderate (-4 MPa) drying, PTS solution moved from the endosperm into the shoot meristem, radicle, and scutellum, but not vice versa. Radicle emergence and elongation proceeded for up to 8 h. With harsh (-150 MPa) drying, PTS concentrated almost exclusively in the radicle. These data illustrate that the endosperm is at least a temporary water storage compartment external to the embryo itself. We speculate that water supplied by the endosperm may be important in reducing the harmful effects of drying during the critical transition period when a germinating seed changes from a desiccation-tolerant to a desiccation-intolerant organism.     

赤霉素与脱落酸对番茄种子萌发中细胞周期的调控

利用细胞流检仪检测番茄（Ｌｙｃｏｐｅｒｓｉｃｏｎ ｅｓｃｕｌｅｎｔｕｍ Ｍｉｌｌ．） ＧＡ－缺陷型、ＡＢＡ－缺陷型和相应的正常品种（野生型）成熟种子胚根尖细胞倍性水平时发现：ＧＡ－缺陷型和野生型种子绝大多数细胞ＤＮＡ 水平为２Ｃ,而ＡＢＡ－缺陷型种子则含有较多的４Ｃ细胞。在标准发芽条件下,ＡＢＡ－缺陷型和野生型种子浸种１ ｄ 后胚根尖细胞ＤＮＡ 开始复制,随后胚根突破种皮而发芽。然而ＧＡ－缺陷型种子除非加入外源ＧＡ,否则既不发生细胞ＤＮＡ 复制,也不发芽。这说明内源ＧＡ 是启动番茄种子胚根尖细胞ＤＮＡ 复制的关键因素,同时也说明番茄根尖细胞ＤＮＡ 复制是种子发芽的必要条件。实验证明：ＡＢＡ 不抑制细胞ＤＮＡ 合成,但阻止Ｇ２ 细胞进入到Ｍ 期。外源ＡＢＡ处理野生型种子与渗控处理结果相似,可以大幅度提高胚根尖４Ｃ／２Ｃ细胞的比例,但抑制种子的最终发芽     

Dormancy in Rice Seed II: THE INFLUENCE OF COVERING STRUCTURES

Most of the dormancy in rice seed can be accounted for by theinhibitory influence of the husk, and most of the residual dormancyafter dehusking can be attributed to the inhibitory influenceof other covering structures—either the pericarp or testa,or both. It is shown that the rate of water absorption is thesame in dormant and non-dormant seeds and that dormant seedsare capable of absorbing sufficient water for germination. Thecovering structures therefore do not cause dormancy by restrictingthe entry of water. Removal of a small area of the husk breaks the dormancy of alarge proportion of the seeds; but for some seeds this treatmentis ineffective whereas removal of the entire husk would breakdormancy. The site of the excision of a small area of the huskcan alter the effectiveness of the treatment: removal of a portionof husk immediately over the embryo is no more effective thanexcising a similar portion nearby, but the removal of part ofthe husk some distance from the embryo is not as effective.Sealing the perforations with paraffin wax has little effectexcept when carried out as soon as possible after the excisionis made, and then only in positions distant from the embryo. Attempts to extract a water-soluble or ether-soluble germinationinhibitor from the husk and other parts of dormant seed or todemonstrate the presence of inhibitors by indirect methods havenot been successful. Nor has it been found possible to extracta water-soluble germination stimulator from seed which has brokendormancy. The implications of these results are discussed.     

A Preliminary Study on Neutral and Acid Inhibitors in the Seed of Kalopanax scptemlobus

The method of extraction, isolation, purification and identification of the neutral and acid inhibitors in the seeds of Kalopanax Septemlobus was given. It is proved that the neutral inhibitor is coumarin, the acid inhibitor is abscisic acid (ABA) by means of paper chromatograph, thin layer chromatograph, high performance liquid chromatograph (HPLC), color reaction and bioassay. The neutral inhibitor can strongly inhibite not only the seed germination of Brassica Chinensis and the radical elongation, but also that of the seed of Kalopanax septemlobus. The study showed that ABA and coumarin exist in the seed coat, endosperm and pericarp of the Kalopanax Septemlobus seed. Both ABA and coumarin can transport from seed cover (pericarp, seed coat)to the interior (endosperm, embryo) as the seeds were stored in refrigerator. In addition, the different results of extracting neutral inhibitor with water, methanol, and alcohol were compared in this paper.     

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.     

Accumulation of Storage Materials,Precocious Germination and Development of Desiccation Tolerance During Seed Maturation in Mustard (Sinapis alba L.)

Under defined environmental conditions (20°C, continuous light of 15 klx) development of mustard seeds from artificial pollination to maturity takes about 60 d. After surpassing the period of embryo cell division and histodifferentiation (12–14d after pollination = dap), the seed enters into a maturation period. The time courses of various physiological, biochemical, and structural changes of embryo and testa during seed maturation were analyzed in detail (dry and fresh mass changes, osmotic and water potential changes, respiration, DNA amplification by endomitosis, total ribosome and polysome formation, storage protein synthesis and accumulation, storage lipid accumulation). In addition to the final storage products protein and lipid, embryo and testa accumulate transiently large amounts of starch within the chloroplasts during early maturation. Concomitantly with the subsequent total breakdown of the starch, the plastids lose most of their internal structure and chlorophyll and shrink into proplastids, typical for the mature seed. At about 30 dap the seeds shift from a desiccation-sensitive to a desiccation-tolerant state and are able then to germinate rapidly upon drying and reimbibition. If isolated from the immature fruit and sown directly on water, the seeds demonstrate precocious germination from about 13 dap onwards. Young seeds (isolated ≦ 38 dap) germinate only after surpassing a lag-phase of several days (after-ripening) during which the embryo continues to accumulate storage protein and lipid at the expense of the surrounding seed tissues. We conclude from these results that the maturing seed represents a rather closed developmental system which is able to continue its development up to successful germination without any specific regulatory influence from the mother plant. Immature seeds are able to germinate without a preceding dehydration treatment, which means that partial or full desiccation does not serve as an environmental signal for reprogramming seed development from maturation to germination. Instead, it is argued that the water relations of the seed are a critical element in the control of maturation and germination: during maturation on the mother plant the embryo is subject to a considerable turgor pressure (of the order of 12 bar) accompanied by a low water potential (of the order of ?12 bar). This turgor permits maturation growth but is subcritical for germination growth. However, upon imbibition in water, the low water potential provides a driving force for a burst of water uptake overcoming the critical turgor threshold and thereby inducing germination.     

HISTOCHEMICAL LOCALIZATION OF ALCOHOL DEHYDROGENASE IN DEVELOPING BEAN SEEDS

Alcohol dehydrogenase activity can be readily localized within bean seeds using the tetrazolium method. In the seed coat, prior to the cotyledon stage of embryo development, staining was seen throughout, especially in the branched parenchyma cells and the integumentary tapetum. As the embryo entered the maturation stage, a gradual decrease in the staining intensity was observed throughout the seed coat. Embryos stained well at all stages of development. In the course of this investigation, a high activity of “nothing dehydrogenase” was observed. Since its activity could be inhibited by methylpyrazole, an ADH inhibitor, it is concluded that the nothing dehydrogenase activity is caused by alcohol dehydrogenase.     

Water uptake and distribution in non-dormant and dormant wild oat (Avena fatua L.) caryopses

The influence of seed coat modification and light quality onwater uptake and distribution in caryopses of dormant and non-dormantlines of wild oat (Avena fatua L.) was determined using NMRmicroimaging. Non-dormant seeds absorbed water more rapidlythan dormant seeds during imbibition on distilled water. Thiseffect was detected first in the embryo-scutellar region (8h) and later in the proximal endosperm (12 h). Cutting the testaand pericarp close to the embryo or scarification with KOH promotedrapid embryo/scutellum hydration and germination. Cutting atthe middle part of the caryopsis did not enhance embryo hydrationnor did it greatly improve germination. The sensitivity of waterdistribution to the phytochrome germination effect was examined.Significant differences in imbibitional water uptake by embryos-scutellumtissue were detected by 18 h following red-light (germinationpromoter) compared with far-red (germination inhibitor) treatment.The results indicated that both the rate and the sequence ofembryo/scutellum hydration were important in initiating germinationin dormant seeds. A refinement of the model that describes waterimbibition in wild oat seeds during the early stages of germinationis discussed. Key words: Water uptake, water distribution, Avena fatua, seed coat modification, light quality, dormant and non-dormant seeds     

The Role of Germination Inhibitors and Oxygen in the Dormancy of the Light-densitive Seed of Betula spp.

Although unchilled, intact seeds of Betula pubescens and B.verrucosa require light for germination, isolated embryos germinateequally well in both light and darkness. An aqueous extract of these seeds has germination-inhibitoryproperties correlated with the presence of a non-fluorescent,single substance. The light requirement of isolated embryosis restored by the inhibitor. When intact seeds are leachedwith water to remove some inhibitor, it is found that the lightrequirement is reduced, short days and single light periodsthen eliciting greater germination than in unleached seeds. It has been found that scratching, pricking, and cutting theseed coat increases the germination of intact seeds in darkness,and that this is probably due to enhanced oxygen entry. Further,it has been found that germination in short days is increasedin oxygen-enriched atmospheres. It has been found that although the inhibitory effect of theseed coat in intact seeds is partially due to the reductionof the oxygen supply to the embryo, a low oxygen concentrationdoes not prevent germination of isolated embryos. Experimentalresults suggest that the inhibitor in the seed coat increasesthe oxygen requirement of the embryo.     

Hypoxia and Imbibition Injuries to Aging Seeds

The development of hypoxia and primary injuries were examined during the imbibition of aging pea seeds (Pisum sativum L., cv. Nemchinovskii). The distribution of air-dry pea seeds by their room-temperature phosphorescence revealed the presence of two fractions (I and II) in a seed lot with 72% germinability and three fractions (I, II, and III) in a seed lot with 50% germinability. The water uptake during imbibition was slower in the fraction I seeds than in the fraction-II seeds. The fraction-I seeds produced normal seedlings, whereas the fraction-II seeds either produced seedlings with morphological defects (abnormal) or did not germinate at all. The fraction-III seeds were all dead. The phosphorescence of endogenous porphyrins, emitted only at low O₂ content, was measured after 20-h seed imbibition. The fraction-I seeds emitted no discernible phosphorescence. The fraction-II comprised highly phosphorescent seeds incapable of radicle protrusion and moderately phosphorescent seeds producing abnormal seedlings. The fraction-II seeds experienced hypoxia during the imbibition because of rapid oxygen consumption by the embryo and restrictions to O₂ diffusion imposed by the seed coat. In the fraction-I seeds, the rate of oxygen consumption by the embryo was slower and the seed coat resistance to oxygen diffusion was lower than in the fraction-II seeds. Therefore, hypoxia did not arise in the fraction-I seeds. The submergence of seeds in water caused lethal injuries. The imbibition of seeds without any contact with water caused no lethal damages but did not reduce the percentage of seeds dying of hypoxia. A slow imbibition of seeds in the media containing either an osmoticum (PEG) or an inhibitor of aquaporin channels (p-chloromercuribenzoate) prevented the lethal injuries at early stages of seed hydration and retarded the appearance of oxygen deficiency in fraction-II seeds. Different rates of water uptake by fraction-I and fraction-II seeds were controlled by permeability of cell membranes rather than by permeability of seed coat. It is proposed that low permeability of plasma membranes to water in fraction-I seeds results from the predominantly closed aquaporin channels, whereas a higher permeability of weak seeds (fraction II) is due to open channels.