Role of seed coat in imbibing soybean seeds observed by micro-magnetic resonance imaging

Background and Aims

Imbibition of Japanese soybean (Glycine max) cultivars was studied using micro-magnetic resonance imaging (MRI) in order to elucidate the mechanism of soaking injury and the protective role of the seed coat.

Methods

Time-lapse images during water uptake were acquired by the single-point imaging (SPI) method at 15-min intervals, for 20 h in the dry seed with seed coat, and for 2 h in seeds with the seed coat removed. The technique visualized water migration within the testa and demonstrated the distortion associated with cotyledon swelling during the very early stages of water uptake.

Key Results

Water soon appeared in the testa and went around the dorsal surface of the seed from near the raphe, then migrated to the hilum region. An obvious protrusion was noted when water reached the hypocotyl and the radicle, followed by swelling of the cotyledons. A convex area was observed around the raphe with the enlargement of the seed. Water was always incorporated into the cotyledons from the abaxial surfaces, leading to swelling and generating a large air space between the adaxial surfaces. Water uptake greatly slowed, and the internal structures, veins and oil-accumulating tissues in the cotyledons developed after the seed stopped expanding. When the testa was removed from the dry seeds before imbibition, the cotyledons were severely damaged within 1·5 h of water uptake.

Conclusions

The activation of the water channel seemed unnecessary for water entry into soybean seeds, and the testa rapidly swelled with steeping in water. However, the testa did not regulate the water incorporation in itself, but rather the rate at which water encountered the hypocotyl, the radicle, and the cotyledons through the inner layer of the seed coat, and thus prevented the destruction of the seed tissues at the beginning of imbibition.Key words: Dry seeds, Glycine max, MRI, seed coat, soaking injury, soybean, testa, role of inner layer of seed coat, water uptake     

Water entry for the black locust (<Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> L.) seeds observed by dedicated micro-magnetic resonance imaging

Water entry at germination for black locust (Robinia pseudoacacia L.) seeds which are known as hard seeds with impermeable seed coat to water, was examined using micro-magnetic resonance imaging (MRI). The MRI apparatus equipped with a low-field (1 T; Tesla) permanent magnet was used, which is open access, easy maintenance, operable and transportable. The excellent point of the apparatus is that T ₁-enhancement of water signals absorbed in dry seeds against steeping free water is stronger than the apparatuses with high-field superconducting magnets, which enabled clear detection of water entry. Water hardly penetrated into the seeds for more than 8 h but approximately 60 % of seeds germinated by incubating on wet filter papers for several days. Hot water treatments above 75 °C for 3 min effectively induced water gap; scarification was 70 % at 100 °C and 75 °C, declined to 15 % at 50 °C and decreased further at room temperature. Water entered into the scarified seeds exclusively through the lens, spread along the dorsal side of the seeds and reached the hypocotyl, whereas water migrated slowly through hilum side to radicle within 3 h.     

Water uptake and distribution in germinating lupine seeds studied by magnetic resonance imaging and NMR spectroscopy

Magnetic resonance imaging (MRI) was used to study temporal and spatial water uptake and distribution in germinating lupine ( Lupinus luteus L.) seeds. During 24 h of imbibition, water was unevenly distributed within the seed and some anatomical parts were more hydrated than others. Water entered the seed through the hilum and micropyle. The embryonic axis was the first to show hydration followed by seed coat and later cotyledons. The changes in water status were characterized by NMR spectroscopy. Analyses of T₂ relaxation times revealed a three-component water proton system (structural, intracellular and extracellular water) in germinating lupine seeds. The data on the components of transverse relaxation time studies indicated the complex exchange processes taking place between water components inside lupine seed over first 2.5 h of hydration, with a distinguished increase in structural water and decrease in other components. This speaks in favor of the high water-absorbing capacity of lupine seeds as related to high protein content. Germination was accompanied by swelling of protein bodies and changes in the organization of stored reserves with gradual disappearance of protein from the cells.     

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.) : V. Water Relations of Imbibition and Germination

The initiation of radicle growth during seed germination may be driven by solute accumulation and increased turgor pressure, by cell wall relaxation, or by weakening of tissues surrounding the embryo. To investigate these possibilities, imbibition kinetics, water contents, and water (Ψ) and solute (ψ_s) potentials of intact muskmelon (Cucumis melo L.) seeds, decoated seeds (testa removed, but a thin perisperm/endosperm envelope remains around the embryo), and isolated cotyledons and embryonic axes were measured. Cotyledons and embryonic axes excised and imbibed as isolated tissues attained water contents 25 and 50% greater, respectively, than the same tissues hydrated within intact seeds. The effect of the testa and perisperm on embryo water content was due to mechanical restriction of embryo swelling and not to impermeability to water. The Ψ and ψ_s of embryo tissues were measured by psychrometry after excision from imbibed intact seeds. For intact or decoated seeds and excised cotyledons, Ψ values were >−0.2 MPa just prior to radicle emergence. The Ψ of excised embryonic axes, however, averaged only −0.6 MPa over the same period. The embryonic axis apparently is mechanically constrained within the testa/perisperm, increasing its total pressure potential until axis Ψ is in equilibrium with cotyledon Ψ, but reducing its water content and resulting in a low Ψ when the constraint is removed. There was no evidence of decreasing ψ_s or increasing turgor pressure (Ψ-ψ_s) prior to radicle growth for either intact seeds or excised tissues. Given the low relative water content of the axes within intact seeds, cell wall relaxation would be ineffective in creating a Ψ gradient for water uptake. Rather, axis growth may be initiated by weakening of the perisperm, thus releasing the external pressure and creating a Ψ gradient for water uptake into the axis. The perisperm envelope contains a cap of small, thin-walled endosperm cells adjacent to the radicle tip. We hypothesize that weakening or separation of cells in this region could initiate radicle expansion.     

Water uptake and distribution in germinating tobacco seeds investigated in vivo by nuclear magnetic resonance imaging

The regulation of water uptake of germinating tobacco (Nicotiana tabacum) seeds was studied spatially and temporally by in vivo (1)H-nuclear magnetic resonance (NMR) microimaging and (1)H-magic angle spinning NMR spectroscopy. These nondestructive state-of-the-art methods showed that water distribution in the water uptake phases II and III is inhomogeneous. The micropylar seed end is the major entry point of water. The micropylar endosperm and the radicle show the highest hydration. Germination of tobacco follows a distinct pattern of events: rupture of the testa is followed by rupture of the endosperm. Abscisic acid (ABA) specifically inhibits endosperm rupture and phase III water uptake, but does not alter the spatial and temporal pattern of phase I and II water uptake. Testa rupture was associated with an increase in water uptake due to initial embryo elongation, which was not inhibited by ABA. Overexpression of beta-1,3-glucanase in the seed-covering layers of transgenic tobacco seeds did not alter the moisture sorption isotherms or the spatial pattern of water uptake during imbibition, but partially reverted the ABA inhibition of phase III water uptake and of endosperm rupture. In vivo (13)C-magic angle spinning NMR spectroscopy showed that seed oil mobilization is not inhibited by ABA. ABA therefore does not inhibit germination by preventing oil mobilization or by decreasing the water-holding capacity of the micropylar endosperm and the radicle. Our results support the proposal that different seed tissues and organs hydrate at different extents and that the micropylar endosperm region of tobacco acts as a water reservoir for the embryo.     

桃儿七种子解剖结构及其萌发生长期形态特征

为探明种皮和胚乳是否是限制桃儿七种子萌发的主要因素,利用组织切片和显微技术,对桃儿七种子及其不同萌发期（1、7、14、21、28 d）解剖结构和播种后一定时期内（7~210 d）的植株生长形态进行观察。桃儿七种子由种皮、胚乳和胚构成。种皮包括外种皮和内种皮,外种皮致密规整,由外至内分别为栅状石细胞和表皮层细胞,内种皮由5~6层海绵细胞组成。胚乳占种子体积的绝大部分,包括珠孔胚乳和外胚乳。胚由胚根、胚轴和子叶组成,被致密种皮、多层珠孔胚乳和外胚乳包围。萌发期1~7 d胚根和胚轴开始伸长,7~14 d两片子叶分离,14~21 d胚根突破珠孔胚乳和种皮,21~28 d胚根、胚轴和子叶继续扩张伸长。种子播种210 d后可平均形成3片功能真叶和5条不定根。致密种皮（物理休眠）和多层胚乳（机械休眠）是限制桃儿七种子萌发的两个主要因素。     

A comparative study of water distribution, free radical production and activation of antioxidative metabolism in germinating pea seeds

The aim of this study was to investigate whether there is a relationship between hydration of the embryo axes and cotyledons and the resumption of the oxidative metabolism in both organs of germinating seeds of pea (Pisum sativum L. cv. Piast). Nuclear magnetic resonance (¹H-NMR) spectroscopy and imaging were used to study temporal and spatial water uptake and distribution in pea seeds. The observations revealed that water penetrates into the seed through the hilum, micropyle and embryo axes, and cotyledons hydrate to different extents. Thus, inhomogeneous water distribution may influence the resumption of oxidative metabolism. Electron paramagnetic resonance (EPR) measurements showed that seed germination was accompanied by the generation of free radicals with g₁ and g₂ values of 2.0032 and 2.0052, respectively. The values of spectroscopic splitting coefficients suggest that they are quinone radicals. The highest content of free radicals was observed in embryo axes immediately after emergence of the radicle. Glutathione content decreased during the entire germination period in both embryo axes and cotyledons. A different profile was observed for ascorbate, with significant increases in embryo axes, coinciding with radicle protrusion. Electrophoretic analysis showed that superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and glutathione reductase (GR, EC 1.6.4.2) were present in dry seeds and were activated later during germination, especially in embryo axes. The presence of all antioxidative enzymes as well as low molecular antioxidants in dry seeds allowed the antioxidative machinery to be active as soon as the enzymes were reactivated by seed imbibition. The observed changes in free radical levels, antioxidant contents and enzymatic activities in embryo axes and cotyledons appear to be more closely related to metabolic and developmental processes associated with preparation for germination, and do not correspond directly to the hydration of the tissues.     

The Importance of Genetically Determined Seed Coat Characteristics to Seed Quality in Grain Legumes

Comparisons of five pairs of isogenk lines of peas, differingonly in the A gene for seed coat colour showed that white seeds(genotype aa) imbibed more rapidly than coloured seeds (AA),suffered greater imbibition damage revealed by dead tissue onthe cotyledons, and higher solute leakage. Seed-coat pigmentationwas closely associated with slow water uptake, since when expressionof the A gene was suppressed by the recessive pollens gene,the resulting white seeds {palpal AA) imbibed rapidly. The slowwater uptake by coloured seeds was not due to the restrictionof water entry by the seed coat since the differences in imbibitionrate were maintained when a portion of the seed coat was removedand seeds were imbibed with the exposed cotyledon in contactwith moist filter paper. Imbibition of similarly treated seedsby immersion in polyethylene glycol solutions (1–4%) whichincreased the seed/solution wettability, had little effect onthe water uptake of coloured seeds compared to imbibition inwater whereas that of white seeds increased in the first 10mins imbibition. Poor wettability of the inner surface of colouredseed coats did not therefore explain the slow imbibition ofthese seeds. The white seed coats loosened rapidly during imbibitionwhilst the coloured seed coats remained closely associated withthe cotyledons suggesting that the adherence of the seed coatto the cotyledons and therefore the ease of access of waterbetween the testa and cotyledons determines the rate of imbibition.The rapid water uptake by white-coated seeds and the subsequentimbibition damage may explain the high incidence of infectionof these seeds by the soil-bome fungus Pythhan after 2 d insoil. Improved seed quality and emergence may therefore be achievedby breeding for seed coat characteristics leading to reducedrates of imbibition Pisum sativum, isogenic lines, A gene, seed coat colour, imbibition, imbibition damage, wettability, pollens gene, seed quality, grain legumes     

Effect of tannin content of faba bean (Vicia faba) seed on seed vigour, germination and field emergence

Differences in seed vigour of zero- and high-tannin faba beans were investigated using 25 seed lots of 12 cultivars following earlier reports of poor emergence in the zero-tannin types. Field emergence ranged from 54–96% indicating differences in seed vigour between cultivars all having high laboratory germination (>91%). Seed from zero-tannin accessions with poor emergence had a higher incidence of testa and cotyledon cracking, a smaller percentage of hard seeds, more rapid water uptake, a lower percentage of vital staining of cotyledons and a greater leaching of solutes than high-tannin types. Nevertheless, variation in these characteristics existed between cultivars and lines of both types. Seeds with more cracks in the seed coat and fewer hard seeds imbibed water more rapidly and consequently showed lower levels of vital staining and more cracks in the abaxial surface of the cotyledons. Slower imbibition in polyethylene glycol lessened the incidence of these deleterious characteristics and may provide a practical resolution to the problem of poor field emergence in zero-tannin lines of faba bean with low seed vigour.     

The Damaging Effect of Water on Dry Pea Embryos During Imbibition

When pea seeds were imbibed in water without their seed coats,vital staining revealed that cells on the abaxial surface ofthe cotyledons were dead. No damage occurred on the surfaceof cotyledons when the seeds were imbibed intact, or beneaththe testa when only half of the testa was removed. Cell deathoccurred as a result of rapid water uptake within the first2 min of imbibition, since reducing the rate of imbibition insolutions of Carbowax 4000 lessened the damage. Cell death wasrestricted to the outer layers of the cotyledons; inner tissuesremained alive. These observations supported the hypothesisthat rapid early leakage during imbibition of dry embryos resultedfrom the death of cells caused by the physical disruption ofmembranes. Imbibition damage resulted in reduced respirationand germination, a decline in the rate of food reserve transferfrom the cotyledons to the growing axis, and a lower growthrate in the seedlings produced. Greater sensitivity of embryosto imbibition damage at low temperature, and similarities betweenfeatures of imbibition damage and chilling injury led to thesuggestion that so-called chilling injury is the result of imbibitiondamage rather than the effects of low temperature.     

The Appearance of Lactic Acid in Phaseolus Seeds Germinating under Wet Conditions

Under normal conditions of germination, there is never muchlactate in bean cotyledons, but when beans are germinated underwet conditions lactate accumulates for about 30 h and then slowlydisappears, as radicle emergence begins. Removal of the testaefrom soaked seeds prevents subsequent lactate accumulation.Lactate also appears in beans germinated under nitrogen. Lactate dehydrogenase activity is enhanced in wet or anaerobicconditions. Incubation with pyruvate increased lactate dehydrogenaseby 50 per cent. There was also a temporary accumulation of alcohol in beansgrown under wet conditions. Under wet conditions the production of respiratory carbon dioxidewas particularly slow during the period of lactate accumulation.     

Pepper seed germination assessed by combined X-radiography and computer-aided imaging analysis

A lot of pepper seeds having 87 % germination were subjected to X-ray inspection using a non lethal dose of radiation. Seeds with less than 2.7 % (on the basis of total seed area) of free space area, i.e. the spaces between embryo and endosperm, were classified as highly viable seeds (97–100 % germination) with the lowest level of abnormal seedlings. Seeds X-ray classified as good were subjected to a computerised image analysis to study seed imbibition and radicle elongation. The patterns of seed area increase, chosen as the most accurate indicator of seed swelling, resembled the triphasic curve of water uptake. The first phase was completed at 9 h followed by a second phase that varied widely in time until completion of germination between 52 and 96 h. The proportion of seeds with radicle protrusion between 52–56 h and 64–72 h assessed with the image analysis was significantly higher than that recorded using a conventional germination test. In addition, the rate of increase of seed area during the third phase of imbibition, mostly due to protrusion of the radicle tip and its growth, was highly correlated with the corresponding radicle elongation rate.     

Water uptake in germinating pecan (Carya illinoinensis) seed

• Water uptake is the fundamental and essential requirement for seed germination. Pecan seed has a hard woody endocarp that plays an important role during water uptake.
• To explore water uptake during germination, the spatiotemporal pattern of water and effect of the endocarp were analysed using high-field MRI, dye-tracing, wax blocking and SEM of water uptake.
• Isolated seeds completed water uptake in 8 h while whole seeds required 6 days, hence, cracking the endocarp plays an important role. The hilum is the channel through which water enters the seed, while the remainder of the seed coat consist of cells covered with a waxy layer that act as a barrier to water absorption. The region with the highest water content in pecan seed is the edge of the U-shaped region, and water can progressively diffuse from this U-shaped region into the whole kernel.
• We report a new water absorption stage between phase II and phase III of the triphasic model of water uptake of pecan seeds. Cracking the endocarp changed water distribution in pecan seeds, which may trigger further water absorption and radicle elongation.

     

The Influence of Testa Condition on the Imbibition and Vigour of Pea Seeds

Seed lots of peas with high rates of water uptake which containeda large proportion of seeds with at least one crack in the testaand were low in vigour, as indicated by electrolyte leakageand poor field emergence. Comparisons of the tetrazolium chloridestaining of the cotyledons after imbibition in water and polyethyleneglycol of two seed lots both with and without scarificationshowed that cracks in the testa could cause cell death by increasingthe rate of water uptake. Although the major cause of poor seedvigour appeared to be the incidence of cracks in the testa whichresulted in rapid water uptake, cell death, and solute leakage,the condition of the embryo also influenced the response towater uptake. The practical significance of these observationsto the problems of field emergence and seed production in peasis discussed.     

The Role of the Lens in Seed Imbibition and Seedling Vigour of Sesbania punicea (Cav.) Benth. (Leguminosae: Papilionoideae)

Seeds of Sesbania punicea (Cav.) Benth. are largely impermeable,and the natural, low, percentage which germinate are permeableat the lens. Scarifying seed in acid stimulates germinationand causes various types of damage to the testa, including thelens. Placing seeds in boiling water stimulates germinationby rendering the seeds permeable at the lens. Seeds were mechanicallyscarified at precise sites on the testa to assess influenceof the site and degree of damage on germination. Results indicatethat both the site of initial water entry and the rate of waterentry are important in determining seed and seedling vigour.Damaging the lens is more effective in stimulating germinationand reducing seedling abnormalities than complete excision ofthe lens. This effect can be reversed if seeds from which thelens was excised are allowed to imbibe gradually in inert osmotica.The leguminous lens thus acts as the site of initial water entryand regulates the rate of water uptake, thereby increasing seedand seedling vigour. Sesbania punicea (Cav.) Benth., hardseededness, lens, seed germination, imbibition damage, seedling vigour     

Hormonal regulation of germination and early seedling development in Acer pseudoplatanus (L.)

Summary Dormancy of intact sycamore (Acer pseudoplatanus) seeds was broken by chilling (5°C) for several weeks in moist conditions. Treatment of unchilled seeds with kinetin induced some germination, but gibberellin was ineffective. This stimulation by kinetin was not suppressed by the added presence of abscisic acid during incubation.The chilling requirement of intact seeds was eliminated by removal of the testa, and the naked embryos developed with no morphological abnormalities. During early growth of isolated embryos in the light, two distinct developmental processes were recognised. One involved initial elongation of the radicle accompanied by geotropic curvature and was stimulated by kinetin but not by gibberellin, while the other involved unrolling of the cotyledons, which was accelerated by gibberellin but much less by kinetin. Abscisic acid strongly suppressed both developmental processes when applied alone, inhibited cotyledon expansion in the presence of gibberellin, but failed to overcome the promotory effects of kinetin on radicle growth. Experiments with CCC indicated that under natural conditions the unrolling of the cotyledons is dependent upon endogenous gibberellin. Radicle growth of isolated embryos was unimpaired by incubation in the dark, but cotyledon expansion of water incubated embryos was poor, and although it was accelerated by gibberellin, the responses in all treatments were slower than in the corresponding light grown samples.It is suggested that endogenous cytokinins are primary factors in the initiation of radicle growth, while gibberellins are important in cotyledon expansion. Abscisic acid appears to have an inhibitory role in both processes, and the interactions of these regulators in the control of germination and development are discussed.     

Physical forces in dormancy and germination of xanthium seeds

The germination of seeds of Xanthium pensylvanicum Wallr. occurs in 2 phases, an initial passive phase of water uptake followed by an active phase of growth. These 2 phases have been separated experimentally, and shown to occur similarly in isolated cotyledons and embryonic axes. Measurements of the physical thrust generated by the entire seed and its separate components of cotyledon and axis reveal that non-dormant Xanthium seeds develop more than twice the thrust of dormant seeds, and that this difference develops principally in the second phase of enlargement of the axis. Measurement of the forces required for piercing the testa of these seeds establishes that whereas the thrust developed by non-dormant seed is adequate to cause testa rupture, that developed by dormant seeds is not. It is concluded that the dormancy of Xanthium involves an inadequacy in the embryo for rupture of the testa.     

Water uptake and oil distribution during imbibition of seeds of western white pine (<Emphasis Type="Italic">Pinus monticola</Emphasis> Dougl. ex D. Don) monitored in vivo using magnetic resonance imaging

Dry or fully imbibed seeds of western white pine (Pinus monticola Dougl. ex D. Don) were studied using high-resolution magnetic resonance imaging (MRI). Analyses of the dry seed revealed many of the gross anatomical features of seed structure. Furthermore, the non-invasive nature of MRI allowed for a study of the dynamics of water and oil distribution during in situ imbibition of a single seed with time-lapse chemical shift selective MRI. During soaking of the dry seed, water penetrated through the seed coat and megagametophyte. The cotyledons of the embryo (located in the chalazal end of the seed) were the first to show hydration followed by the hypocotyl and later the radicle. After penetrating the seed coat, water in the micropylar end of the seed likely also contributed to further hydration of the embryo; however, the micropyle itself did not appear to be a site for water entry into the seed. A model that describes the kinetics of the earlier stages of imbibition is proposed. Non-viable pine seeds captured with MRI displayed atypical imbibition kinetics and were distinguished by their rapid and uncontrolled water uptake. The potential of MR microimaging for detailed studies of water uptake and distribution during the soaking, moist chilling (stratification), and germination of conifer seeds is discussed.Electronic Supplementary Material Supplementary material is available for this article if you access the article at . A link in the frame on the left on that page takes you directly to the supplementary material.     

Structures contributing to the completion of conifer seed germination

The variety of interpretations of the origin and role of the tissues surrounding the emerging radicle of conifer seeds prompted us to study changes during germination. The structures contributing to the opening of the seed coat and protrusion of the radicle in pine and spruce seeds were examined using light microscopy and field emission scanning electron microscopy. The opening of the seed coat was mainly mechanical and primarily affected by the enlargement of the imbibed endotesta cells lining the micropylar canal. The hypertrophied nucellar apex, swelling remnants and mucous substances in the micropyle further enhanced the testa splitting which was reinforced by the straightening of the folded nucellar cap. The expansion of the imbibed megagametophyte and the embryo were conducive to the seed coat opening only at a later stage. This was followed almost immediately by the appearance of the root cap showing an early geotropic response typical for taproots and assuring an immediate anchoring of the emerged embryo to the ground. The translucent tissue composed of elongating cell rows preceded and surrounded the protruding radicle and derived mainly from two origins: the micropylar end of the megagametophyte and the root cap. Observations about the origin of the sheath clarify the reasons for previous interpretations about the putative role of the massive embryo root cap in testa splitting and germination. The protective and adhesive function of the sheath is concluded to be essential to the conifer embryos, which present the epigeal type of germination. Received: 7 July 1999 / Accepted: 11 August 1999     

The Role of ABA in the Control of Apple Seed Dormancy Re-appraised by Combined Gas Chromatography-Mass Spectrometry

Measured by GC—MS²—SIR³, endogenous ABA⁴ in embryonicaxes of seeds of Malus pumila L. cv. Golden Delicious decreased8-fold and cotyledon ABA by only 60%, during 10–50 d ofstratification at 5 ?C, after ABA leaching during an initial24 h soaking. During stratification, the percentage germinationof embryos transferred to 17?C showed a significant linear dependenceon log_e of ABA levels in the axes at transfer. Between 50 and70 d, ABA levels increased markedly in axes and testa both ofstratified seeds and seeds allowed to re-dry at 17 ?C afterinitial soaking. The ability of fully stratified axes with elevatedendogenous ABA to germinate indicated that stratification haddecreased their ABA sensitivity. Changes in cotyledon ABA couldnot account for the promotory effect of cotyledons on germinationduring the first 30 d of stratification. Loss of testa inhibitionof germination during stratification was not linked with changesin testa ABA. Stratification markedly increased the sequestrationin the axes of exogenous ABA supplied via the cotyledons. Changesboth in axis ABA levels and sensitivity were thus correlatedwith dormancy release, but subject to modifying control by thecotyledons and testa not involving ABA. Rehydration of driedseeds affected axis ABA later during storage via mechanismsunconnected with dormancy. Key words: ABA, seed dormancy, stratification