Dormancy in Seeds of Charlock: III. OCCURRENCE AND MODE OF ACTION OF AN INHIBITOR ASSOCIATED WITH DORMANCY

The dormancy of charlock seeds appears to be associated withthe presence of an inhibitor which accumulates within the seed.This inhibitor diffuses into the external solution when seedsare placed in water under germination conditions or controlledexperimental conditions. A small quantity of inhibitor diffusesfrom the seed coats but most arises from the embryo. The increasein temperature was measured by comparing the relative growth-ratesof the radicles of excised charlock embryos in water and intest solutions of diffusate. The results suggest that the rateof accumulation of inhibitor in the external solution is controlledby diffusion, and that there is continuous production of inhibitorin the tissues of the embryo which have a low oxgyen tension.The critical concentration of the inhibitor which completelyprevents cell elongation is rapidly attained in seeds whichare dormant. The inhibitor is unlikely to be a mustard oil,such as allylisothio-cyanate.     

Synergism between gibberellic acid and low Pfr levels inducing germination of Kalanchoë seeds

Sown on water, seeds of Kalanchoëbiossfetdiana Poelln. cv. Feuerblute are absolutely light-requiring and show full red/far-red reversibility. In seeds, sown on 2 ×10^-3 M gibberellic acid, red/far-red reversibility disappears and both short red and far-red irradiations induce germination. Gibberellic acid alone does not induce germination, but it increases the physiological activity of P_fr to the extent, that the low P_fr level obtained by far-red irradiation becomes very effective. The synergism between gibberellic acid and far-red light appears after a two-day incubation; period. The nature of this lag phase was examined by measuring both germination and uptake of labelled gibberellic acid in intact seeds and seeds with a punctured seed coat. The lag phase was shown to be independent of the uptake kinetics of gibberellic acid and allows development to a specific stage, necessary for germination after phytochrome-phototransformation. The kinetics of the uptake of gibberellic acid by intact seeds and embryos of intact seeds are different. In intact seeds most of the gibberellic acid is retained in the seed coat; only a small fraction actually penetrates to the embryo where it can exert its physiological activity.     

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.     

准噶尔山楂种壳、种皮、种胚特性与种子休眠的关系

以准噶尔山楂种子为试验材料,检测其种壳和种皮的透水性及超微结构、种胚休眠特性及种子浸提液的抑制作用。结果表明：（1）准噶尔山楂种子具有胚休眠特性,种壳存在一定的机械束缚和透水障碍,存在内源抑制物质是引起准噶尔山楂种子休眠的主要原因。（2）酸蚀处理能使种壳表面结构破损,角质层脱落,种壳变薄,栅栏组织结构和细胞排列未发生变化,种孔增大,种子表面出现裂缝,但并不影响种子生活力。（3）准噶尔山楂种子甲醇浸提液的生物测定结果说明,准噶尔山楂种子的种壳、种皮和种胚均含有内源抑制物质,不同部位浸提液对白菜种子的发芽率、根长和苗高的抑制作用不同。     

Effect of chilling and gibberellic acid on growth potential of excised embryos of Ruellia humilis

Summary Dormancy in intact seeds of Ruellia humilis is broken by chilling or by treatment with gibberellic acid (GA₃). Embryos are nondormant and will grow when the seed coat is removed completely. Embryos from chilled or GA₃-treated seeds have more growth potential than embryos from nontreated seeds. Dormancy is ascribed to the mechanical restriction of the embryo by the seed coat. Chilling and treatment with GA₃ break dormancy by increasing the expansive force of the embryo; thus, chilled or GA₃-treated embryos exert enough expansive force to break through the seed coat, whereas nontreated embryos do not.This work was supported by an NIH Biomedical Sciences Support Grant to the University of Kentucky.     

Seed anatomy and water uptake in relation to seed dormancy in Opuntia tomentosa (Cactaceae, Opuntioideae)

BACKGROUND AND AIMS: There is considerable confusion in the literature concerning impermeability of seeds with 'hard' seed coats, because the ability to take up (imbibe) water has not been tested in most of them. Seeds of Opuntia tomentosa were reported recently to have a water-impermeable seed coat sensu lato (i.e. physical dormancy), in combination with physiological dormancy. However, physical dormancy is not known to occur in Cactaceae. Therefore, the aim of this study was to determine if seeds of O. tomentosa are water-permeable or water-impermeable, i.e. if they have physical dormancy. METHODS: The micromorphology of the seed coat and associated structures were characterized by SEM and light microscopy. Permeability of the seed-covering layers was assessed by an increase in mass of seeds on a wet substrate and by dye-tracking and uptake of tritiated water by intact versus scarified seeds. KEY RESULTS: A germination valve and a water channel are formed in the hilum-micropyle region during dehydration and ageing in seeds of O. tomentosa. The funicular envelope undoubtedly plays a role in germination of Opuntia seeds via restriction of water uptake and mechanical resistance to expansion of the embryo. However, seeds do not exhibit any of three features characteristic of those with physical dormancy. Thus, they do not have a water-impermeable layer(s) of palisade cells (macrosclereids) or a water gap sensu stricto and they imbibe water without the seed coat being disrupted. CONCLUSIONS: Although dormancy in seeds of this species can be broken by scarification, they have physiological dormancy only. Further, based on information in the literature, it is concluded that it is unlikely that any species of Opuntia has physical dormancy. This is the first integrative study of the anatomy, dynamics of water uptake and dormancy in seeds of Cactaceae subfamily Opuntioideae.     

Control of Seed Germination by Abscisic Acid: I. Time Course of Action in Sinapis alba L

The germination process of mustard seeds (Sinapis alba L.) has been characterized by the time courses of water uptake, rupturing of the seed coat (12 hours after sowing), onset of axis growth (18 hours after sowing), and the point of no return, where the seeds lose the ability to survive redesiccation (12 to 24 hours after sowing, depending on embryo part). Abscisic acid (ABA) reversibly arrests embryo development at the brink of radicle growth initiation, inhibiting the water uptake which accompanies embryo growth. Seeds which have been kept dormant by ABA for several days will, after removal of the hormone, rapidly take up water and continue the germination process. Seeds which have been preincubated in water lose the sensitivity to be arrested by ABA after about 12 hours after sowing. This escape from ABA-mediated dormancy is not due to an inactivation of the hormone but to a loss of competence to respond to ABA during the course of germination. The sensitivity to ABA can be restored in these seeds by redrying. It is concluded that a primary action of ABA in inhibiting seed germination is the control of water uptake of the embryo tissues rather than the control of DNA, RNA, or protein syntheses.     

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.     

红松种子休眠与种皮的关系

本文探讨红松(Pinus koraiensis)种子休眠与其种皮之间的关系。夹破中种皮后,种子萌发率很低。在离体胚培养基中外加 ABA 及经 ABA 溶液浸泡种子的萌发实验表明,ABA也不是导致休眠的关键因素。试验确认红松种子存在透气障碍,即中、内种皮对氧气的进入都有阻碍作用。经低温砂藏后,种皮的阻碍作用明显减小。种皮的透气性障碍可能是诱导休限的主导因素。     

血皮槭种子休眠机制研究

利用抑制物生物测定法和酸蚀技术研究了血皮槭种子休眠的原因。血皮槭种子吸水是一个非常缓慢的过程,在140 h以后种子含水量才能达到68%左右。酸蚀处理种子3 h,虽然没有加快种子的吸水速率,但能较好得使果皮变薄,也不影响种子的生活力。种子的各部位（果皮、种皮、子叶、胚根）均含有抑制物质,对小白菜种子的发芽率及胚根生长有很强的抑制作用,子叶各种处理水浸提液的抑制作用最强,果皮和种皮次之。血皮槭种子休眠主要由种壳机械障碍和种胚生理休眠两重因素导致,因此如何克服致密果壳而使激素能接触生理休眠的种胚是打破其种子休眠的关键技术。     

Protein Synthesis in Embryos of Dormant and Germinating Agrostemma githago L. seeds

The time course of protein synthesis in embryos of dormant and afterripened Agrostemma githago seeds was studied. In embryos of afterripened geminating seeds, protein synthesis increased in three successive stages: (a) concurrent with swelling; (b) during the lag phase between the completion of water uptake and the onset of growth; and (c) immediately after protrusion through the seed coat. Embryos of dormant seeds showed the first increase but not the second unless dormancy was broken by imbibition at 4°C. This indicates that dormancy affects processes prior to the onset of growth. The third increase was largely due to higher oxygen availability after the rupture of the seed coat and not to actual growth. It could also be elicited in dormant embryos by isolating them from the seeds.

Electrophoretic analysis of the newly synthesized proteins demonstrated that the patterns of dormant and afterripened embryos became significantly different in both axes and cotyledons only just prior to the onset of axis elongation. Thereafter, the differences became larger.

When afterripened or dormant seeds were transferred from a low, germination-permitting to a high, germination-inhibiting temperature, the seeds germinated at the high temperature if they had completed the lag phase to a sufficient extent at the low temperature. This shows that the processes during the lag phase were inhibited by the high temperature while the onset of growth was not affected.

     

桃儿七种子休眠解除过程中细胞壁代谢及种皮超微结构的变化北大核心CSCD

为探究低温层积过程中桃儿七种子细胞壁代谢及种皮超微结构与休眠解除的内在联系,该研究通过低温层积解除桃儿七种子休眠,分析休眠解除过程中种子不同部位细胞壁组分及相关代谢酶的变化,同时利用扫描电镜对种皮的超微结构进行观察。结果表明,(1)桃儿七种皮主要由角质层、栅状石细胞层及海绵组织层3层构成,在层积过程中,种皮内部的海绵组织逐步疏松膨胀,种皮表面破损加剧;(2)种子不同部位的细胞壁组分具有明显差异,整个层积过程中,种胚、种皮和胚乳中的纤维素含量均在层积中期(45 d和60 d)降至最低,3个部位的纤维素酶活性在层积中期对应升高;种胚和种皮内的半纤维素含量均在层积中期显著下降,种皮中甘露聚糖酶活性和木糖苷酶活性在层积中期时相应达到最大;3个部位的果胶含量均在层积后期(75 d和90 d)时显著下降,而种皮和胚乳中多聚半乳糖醛缩酶活性也在层积后期相应升高;(3)种胚和胚乳内过氧化物酶活性在层积75 d和90 d时明显下降,而SOD活性在此时显著上升。(4)种子不同部位3种木质素单体的组成比例具有明显区别,同时3种木质素单体含量均随层积时间的延长而显著降低,且胚乳和种皮中的S-木质素含量对种子萌发存在显著的负向影响关系。研究认为,在低温层积过程中,桃儿七种子内细胞壁组分纤维素、半纤维素及木质素的逐步酶解,活性氧作用下的细胞壁松弛以及海绵组织层的疏松膨胀和种皮的破裂,破坏了细胞壁的刚性结构,促使种子机械束缚力降低,吸水性能提高、胚根生长能力增强,最终导致其休眠解除。     

Germination des semences duChenopodium bonus- henricus. Recherche du mécanisme de l’inaptitude á la germination des semences récoltées á une altitude élevée

A decreased germination capacity of the seeds ofChenopodium bonus- henricut collected at high altitude is not due to an embryo dormancy. It is caused by a seed coat inhibition. The seed coats contain large amount of strongly oxidisable phenolic compounds which deprive the embryo of oxygen.     

The Role of Water Uptake in the Transition of Recalcitrant Seeds from Dormancy to Germination

In recalcitrant seeds of horse chestnut (Aesculus hippocastanum L.) maintaining a high water content during winter, dormancy is determined by the presence and influence of the seed coat, while the axial organs of the embryos excised from these seeds are not dormant. Such axial organs were capable for active water uptake and rapid fresh weight increase, so that their fresh weights exceeded those in intact seeds at the time of radicle protrusion. Fructose plays an essential role in the water uptake as a major osmotically active compound. ABA interferes with the water uptake by the axial organs and thus delays the commencement of their growth. The manifestation of seed response to ABA during the entire dormancy period indicates the presence of active ABA receptors and the pathways of its signal transduction. The content of endogenous ABA in the embryo axes doubled in the middle of dormancy period, which coincided with a partial suppression of water uptake by the axes. During seed dormancy release and imbibition before radicle protrusion, the level of endogenous ABA in axes declined gradually. Application of exogenous ABA can imitate dormancy by limiting water absorption by axial organs. Fusicoccin A (FC A) treatment neutralized completely this ABA effect. Endogenous FC-like ligands were detected in the seed axial organs during dormancy release and germination. Apparently, endogenous FC stimulates water uptake via the activation of plasmalemmal H⁺-ATPase, acidification of cell walls, their loosening, and turgor pressure reduction. FC can evidently counteract the ABA-induced suppression of water uptake by controlling the activity of H⁺-ATPase. It is likely that, in dormant intact recalcitrant seeds, axial organs, maintaining a high water content, are competent to elevate their water content and to start their preparation for germination under the influence of FC when coat-imposed dormancy becomes weaker.     

SEEDS OF KOSTELETZKYA VIRGINICA (MALVACEAE): THEIR STRUCTURE,GERMINATION, AND SALT TOLERANCE. I. SEED STRUCTURE AND GERMINATION

Dormancy of Kosteletzkya virginica (L.) Presl. seeds is primarily due to the impermeability of the seed coat to water. The impermeable structure is assumed to be, in other Malvaceae, the palisade layer of the seed coat. The percentage of seeds capable of imbibition and germination increased with increasing time of storage at low temperatures, but the release from dormancy was not accompanied by decreased seed coat resistance to pressure. Under natural conditions, mechanical damage to the seed coat due to changes in temperature and/or abrasion may render the seeds water permeable. It is not clear what causes water permeability during storage under laboratory conditions. During seed maturation and drying, the inner epidermis of the tegmen partly separates from the rest of the seed coat and an air space, which makes the seed buoyant, is formed around the region of the chalazal cleft. The optimal temperature for germination of K. virginica seeds is between 28 and 30 C in light or darkness.     

Seed Coat Dormancy in Two Species of Grevillea(Proteaceae)

The role played by the seed coat in seed dormancy of Grevillealinearifolia(Cav.) Druce and G. wilsonii(A. Cunn.) was testedby a series of manipulations in which the seed coat was dissectedand removed, dissected and returned to the decoated seed, ordissected, removed and given a heat shock, and returned to thedecoated seed. Germination of intact seeds of both species wasalso examined after exposure to heat shock, smoke, or heat shockand smoke combined. Water permeability of the seed coat wasinvestigated by examining imbibition. For intact seeds, virtuallyno germination occurred under any treatment (G. wilsonii), orgermination was increased by exposure to either heat or smoke(G. linearifolia). Removal of the seed coat led to germinationof all decoated seeds for G. linearifolia, or a proportion ofdecoated seeds for G. wilsonii. Inclusion of smoked water inthe incubation medium led to a higher proportion of decoatedseeds germinating for G. wilsonii. Returning the seed coat,either with or without heat shock to the seed coat, did notsignificantly affect germination in either species. Seed coatswere permeable to water in both species. For the two Grevilleaspecies, there were different dormancy mechanisms that werecontrolled by the seed coat (G. linearifolia) or by both theseed coat and embryo (G. wilsonii). Copyright 2000 Annals ofBotany Company Grevillea linearifolia, Grevillea wilsonii, dormancy, seed coat dormancy, seed coat permeability, smoke, heat shock, germination     

The Effect of Oxygen on Endogenous Cytokinin Levels and Germination of Leucadendron daphnoides Seed

The soybean callus assay was used to study the effect of high oxygen tensions on the cytokinin levels of Leucadendron daphnoides Meisn. seed, where dormancy is apparently due to the restricting effect of the seed coat on oxygen diffusion to the embryo. High oxygen tensions led to a six-fold increase in germination compared to seed incubated in air and resulted in significant increases in butanol soluble cytokinins prior to visible germination. It is suggested that the primary effect of oxygen is to increase the rate of respiration and thus, to provide the energy required for the synthesis of butanol soluble cytokinins which leads to cotyledon expansion and subsequent radicle elongation. Present indications are that untreated seeds remain dormant due to low concentrations of butanol soluble cytokinins in their embryos.     

Anatomical and chemical characteristics of the seed coat of <i>Medicago sativa</i> L. (alfalfa) cv. Baralfa 85 seeds and their association with seed dormancy

Seeds of alfalfa (Medicago sativa L.) can exhibit seedcoat imposed dormancy, which produces hard seeds within a seed lot. These seeds do not germinate because they do not imbibe water due to a barrier to water entry in the seed coat. The aim of this work was to analyze the anatomical and chemical characteristics of the testa of alfalfa seeds with respect to water permeability levels. The anatomy of seeds of the cv. Baralfa 85 was studied and structural substances, polyphenols, tannins and cutin present in the testa of seeds of different water permeability levels were determined. The anatomical characteristics of the seed coat and the proportions of components were found to determine the permeability level of the seed coat, an aspect that is associated with the physical seed dormancy level. Anatomically, increased thickness of the testa was associated with a lower permeability level. The difference may be attributed to the variation in cuticle thickness, length of macrosclereids and thickness of the cell wall, and presence and development of osteosclereids. From the physiological and chemical points of view, the mechanism of physical dormancy of the testa is explained by a greater amount of components that repel water and cement the cell wall, such as polyphenols, lignins, condensed tannins, pectic substances, and a lower proportion of cellulose and hemicellulose.     

Investigating seed dormancy in cotton (Gossypium hirsutum L.): understanding the physiological changes in embryo during after‐ripening and germination

• The dormancy of seeds of upland cotton can be broken during dry after‐ripening, but the mechanism of its dormancy release remains unclear.
• Freshly harvested cotton seeds were subjected to after‐ripening for 180 days. Cotton seeds from different days of after‐ripening (DAR) were sampled for dynamic physiological determination and germination tests. The intact seeds and isolated embryos were germinated to assess effects of the seed coat on embryo germination. Content of H₂O₂ and phytohormones and activities of antioxidant enzymes and glucose‐6‐phosphate dehydrogenase were measured during after‐ripening and germination.
• Germination of intact seeds increased from 7% upon harvest to 96% at 30 DAR, while embryo germination improved from an initial rate of 82% to 100% after 14 DAR. Based on T₅₀ (time when 50% of seeds germinate) and germination index, the intact seed and isolated embryo needed 30 and 21 DAR, respectively, to acquire relatively stable germination. The content of H₂O₂ increased during after‐ripening and continued to increase within the first few hours of imbibition, along with a decrease in abscisic acid (ABA) content. A noticeable increase was observed in gibberellic acid content during germination when ABA content decreased to a lower level. Coat removal treatment accelerated embryo absorption of water, which further improved the accumulation of H₂O₂ and changed peroxidase content during germination.
• For cotton seed, the alleviation of coat‐imposed dormancy required 30 days of after‐ripening, accompanied by rapid dormancy release (within 21 DAR) in naked embryos. H₂O₂ acted as a core link between the response to environmental changes and induction of other physiological changes for breaking seed dormancy.

     

Seed coat structure and oxygen availability control lowtemperature germination of melon (Cucumis melo) seeds

The involvement of the seed coat in low-temperature germination of melon seeds was examined in two accessions differing in their ability to germinate at 14°C: Noy Yizre'el (a cold-sensitive cultivar) and Persia 202 (a cold-tolerant breeding line). Decoating resulted in full germination of Noy Yizre'el at 14°C, but splitting the coat increased germination only partially. Thus, the inhibition of Noy Yizre'el germination at 14°C is not due to physical constraint on radicle protrusion. At 25°C, seeds of both accessions submerged in water or agar germinated fully as long as the hilum aperture remained uncovered. Submerging the whole seed, or covering the hilum with lanolin, strongly depressed germination of Noy Yizre'el but not of Persia 202. Accessions differed in germination response to decreasing O₂ concentration, with Noy Yizre'el showing higher sensitivity to hypoxia. These differences were correlated with differences in seed coat structure as well as in embryo sensitivity to hypoxia. Intercellular spaces in the outer layer of the seed coat were evident in the more tolerant Persia 202, while in the sensitive Noy Yizre'el this layer was completely sealed. Sensitivity to hypoxia increased at 15°C as compared with 25°C, the increase being greater in Noy Yizre'el. It is proposed that the seed coat-imposed dormancy at low temperature in Noy Yizre'el is the combined result of more restricted oxygen diffusion through the seed coat and a greater embryo sensitivity to hypoxia, rather than to physical constraints of radicle break-through or impairment of imbibition.