Studies on the Germination of Cereals: 2. Factors Determining the Germination Behaviour of Wheat Grains during Maturation

A comparative study has been made of the germination behaviourof a white-grained and a red-grained wheat variety throughoutthe course of maturation. Although freshly harvested wheat grains do not germinate duringthe early stages of ripening, the embryo will develop undergermination conditions when it is completely exposed, or theepidermis of the pericarp only is removed. At the harvest-ripestage the white grains germinate rapidly, but the emergenceof the embryo is considerably delayed in the red grains. Thevarietal difference is due to the presence of the covering layersand becomes increasingly apparent in grains which have beenprematurely harvested, between 3 and 6 weeks after anthesis,and then subjected to desiccation. This effect of the coveringlayers on embryo development does not appear to be related totheir permeability to water or oxygen. The behaviour of thetwo wheat varieties is thus determined by the mechanical propertiesof the covering layers, and especially the pericarp. These delaygermination in the red grains until there has been an increasein the water absorbing power of the imbibed embryo.     

Studies on the Germination of Cereals: 4. The Oxygen Requirement for Germination of Wheat Grains during Maturation

The oxygen required by the wheat embryo, for germination atdifferent stages of maturation, has been determined by the numberof grains with the embryo exposed which germinated in differentoxygen concentrations. Some embryos required only 0.5 per cent.,and the majority required less than 5 per cent., but there wasalso a proportion which required a concentration approachingthe level in air. There were more embryos in the last categoryin the grains of a red variety than in those of a white varietybut this did not account for the varietal difference in germinationwhen the grains were intact. The effect of the covering layers was also determined in eachatmosphere by the difference in germination between intact grainsand grains with the embryo exposed. The covering layers weresufficiently impermeable to prevent germination in both varietieswhen the external oxygen concentration was very low; when theconcentration was increased from 5 to 60 per cent., however,the inhibiting effect of the covering layers was reduced inthe white variety but not in the red. The delayed germinationof the red grains was therefore attributed to some factorotherthan the impermeability of the covering layers to oxygen.     

Structure and water absorption of different parts of the rose achene pericarp during imbibition

Rose achene imbibition is difficult. Comparison of sulphuric acid treated, Cross and Beavanś reagent (C. B. reagent) treated seeds with untreated seeds showed that, during imbibition, water is essentially found in the most external pericarp layer. Inner layers appear less permeable. It is possible to distinguish three layers in the pericarp by taking into account results from imbibition assays, X-ray radiographies and scanning electron microscopic observations. They can be interpreted as representing the epi-, meso- and endocarp.     

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     

The Role of Imbibition Damage in Determining the Vigour of White and Coloured Seed Lots of Dwarf French Beans (Phaseolus vulgaris)

Seeds of dwarf French beans showed evidence of damage alterimbibition in water in the form of dead tissue and high soluteleakage. Damage was more extensive in seeds of a cultivar withwhite testae (Tenderette) both at 20?C and with the initial6 h imbibition at 4?C where damage was more severe, whereasa cultivar with brown testae (Provider) was damaged only alterthe 4?C treatment. Tenderette imbibed more rapidly than Providerin both imbibition treatments. The significant correlationswithin each cultivar of both solute leakage (positive) and theextent of living tissue (negative) with the rate of water uptake,and the reduced damage when seeds imbibed slowly in PEG indicatedthat this was imbibition damage due to rapid water uptake. Differencesin the rates of water uptake were also observed in sand at 15%and 20% mc where rapid imbibition was associated with reducedgermination and the production of more abnormal seedlings. Atemperature of 4?C during the first 6 h germination in sandat 15% mc also reduced germination, particularly in Tcnderette.It is suggested that the more rapid water uptake by white seededcultivars of dwarf French beans and their greater susceptibilityto imbibition damage are responsible for their low vigour. Key words: Imbibition damage, vigour, dwarf French beans     

Regulation of dormancy in barley by blue light and after-ripening: effects on abscisic acid and gibberellin metabolism

White light strongly promotes dormancy in freshly harvested cereal grains, whereas dark and after-ripening have the opposite effect. We have analyzed the interaction of light and after-ripening on abscisic acid (ABA) and gibberellin (GA) metabolism genes and dormancy in barley (Hordeum vulgare 'Betzes'). Analysis of gene expression in imbibed barley grains shows that different ABA metabolism genes are targeted by white light and after-ripening. Of the genes examined, white light promotes the expression of an ABA biosynthetic gene, HvNCED1, in embryos. Consistent with this result, enzyme-linked immunosorbent assays show that dormant grains imbibed under white light have higher embryo ABA content than grains imbibed in the dark. After-ripening has no effect on expression of ABA biosynthesis genes, but promotes expression of an ABA catabolism gene (HvABA8'OH1), a GA biosynthetic gene (HvGA3ox2), and a GA catabolic gene (HvGA2ox3) following imbibition. Blue light mimics the effects of white light on germination, ABA levels, and expression of GA and ABA metabolism genes. Red and far-red light have no effect on germination, ABA levels, or HvNCED1. RNA interference experiments in transgenic barley plants support a role of HvABA8'OH1 in dormancy release. Reduced HvABA8'OH1 expression in transgenic HvABA8'OH1 RNAi grains results in higher levels of ABA and increased dormancy compared to nontransgenic grains.     

Endosperm and pericarp involvement in the supercooling of imbibed lettuce seeds

Fully hydrated lettuce (Lactuca sativa L.) seeds showed dual freezing exotherms (−9 and −18°C), even after 10 hours imbibition. Only the −9°C exotherm was observed in seeds imbibed for 20 hours, but without external nucleation, all water in the embryo supercooled. Results indicate that the endosperm acts as a barrier to ice propagation. Other experiments suggest that the pericarp may also protect the embryo under certain freezing conditions.     

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     

Studies on the Germination of Cereals: I. The Germination of Wheat Grains in the Ear during Development, Ripening, and After-ripening

The ability of individual grains to germinate in the ears ofa red and a white wheat variety has been determined at differentperiods after anthesis, and at different moisture contents,before the stage of full maturity. No grains germinated while active growth was taking place, butafter desiccation during ripening, 88·5 per cent. ofthe white grains and 7 per cent. of the red grains were ableto germinate in the ear; the percentage germination of the redgrains increased to 83 per cent., when further desiccation occurredduring the first 5 weeks of after-ripening, but some grainsin the basal spikelets of the ears of both varieties failedto germinate until they had been subjected to the same desiccationfrom 13 to 23 weeks after anthesis. The ability of the grains to germinate has been correlated withtheir desiccation at different stages during maturation, andthe effect of certain factors, which inhibit the germinationof immature grains, are discussed in relation to varietal differencesin the colour of the grains and their position in the ear.     

Water uptake and splitting of wild oat caryopsis

Imbibition and germination experiments were conducted on the caryopses of wild oats (Avena fatua L.). The embryo envelopes, pericarp and aleurone layer, which completely cover the embryo-endosperm, do not form barriers against water uptake. The initial uptake of water is passive and the water moves across the pericarp with ease as it contains cracks; it is, however, transported across the aleurone layer through its cell walls into the endosperm and embryo of the caryopsis. The starchy endosperm enlarges due to water uptake causing the pericarp to rupture, thus exposing the aleuronelayer-covered seed. The aleurone layer is structurally heterogenous consistings of radially compressed irregular cells and cuboidal or radiallys tretched cells; the latter contains thicker walls. The former type is present along the abaxial side of the embryo and in the crease on the adaxial side of the caryopsis; the latter type covers the endosperm. The physical distention of the endosperm due to water uptake causes the rupture of the pericarp and the aleurone layer, and facilitates the emergence of the radicle and coleorhiza of the embryo during caryopsis germination.     

Herbicidal Effects on Water Uptake of Seeds of Selected Weed Species

Seeds of yellow foxtail, johnsongrass, pigweed and quackgrass were investigated to determine the effects of certain herbicides on water uptake by these species. Seeds of johnsongrass showed slight increase in water uptake over the 20-h period when treated with 1, 5, 10, and 500 mg/1 of 2,4-D. Tordon caused a most pronounced increase in water uptake at 0.1 and 1 mg/l as compared with the controls. Johnsongrass gave indications of either increases or decreases in water uptake when treated with herbicides during the 20-h period, but showed no effects during the 10-h period. Dormant seeds of yellow foxtail indicated an enhancement in water uptake at all 2,4-D concentrations during the 20 h period. The effects which the different herbicides had upon the uptake of water in yellow foxtail seeds seemed to be directly proportional to increase in imbibition time. In all cases, more water was imbibed after 20 h over that observed at 10 h. During the 10-h imbibition period, pigweed seeds treated with 2,4-D was noted to have a slight stimulating effect on water uptake at all concentrations employed except 5 mg/l, which indicated a depressing effect. It was also observed that 2,4-D caused a significant stimulating effect on water uptake of pigweed seeds over the 20-h period as compared with the controls. A stimulation in the total quantity of water imbibed after 20 h was noted at dacthal concentrations below 100 mg/l whereas at the 500 mg/l, a slight depression in water imbibition was observed. At dalapon concentrations of 1 to 100 mg/l, a marked stimulation in total water imbibed by quackgrass seeds was shown during the 10-h period.     

绞股蓝果实的发育解剖学研究

应用石蜡切片和扫描电镜的方法,观察了绞股蓝果实与种子的发育过程,结果表明:花后12 d左右,绞股蓝合子开始进行第一次分裂,极核分裂在花后6～7 d,核型胚乳在胚的发育中被吸收,胚的发育为茄型,外珠被呈网格状花纹。假果,表面有蜡质,果壁可明显分为3层,内侧细胞后期解体,种子倒三角形,表面瘤状突起。脱落花果的结构表现为珠被萎缩和胚发育受阻。     

No influence of the embryonic axis on the development of diamine oxidase in pea cotyledons

A reexamination has been made for the supposed regulation of pea (Pisum sativum cv Alaska) cotyledonary diamine oxidase (EC 1.4.3.6) activity by the embryonic axis. When dry cotyledons from which the embryo and testa have been removed surgically are imbibed by soaking in water, there is little increase of the enzyme activity during subsequent incubation on filter paper. However, if the dry cotyledons are imbibed and maintained on filter paper from the first, the increase of the enzyme activity is similar to that in the intact seedling. Thus, rapid imbibition of the isolated dry cotyledons is responsible for repression of enzyme development, and a role for the axis need not be invoked.     

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.     

Physiological characteristics and related gene expression of after‐ripening on seed dormancy release in rice

After‐ripening is a common method used for dormancy release in rice. In this study, the rice variety Jiucaiqing (Oryza sativa L. subsp. japonica) was used to determine dormancy release following different after‐ripening times (1, 2 and 3 months). Germination speed, germination percentage and seedling emergence increased with after‐ripening; more than 95% germination and 85% seedling emergence were observed following 1 month of after‐ripening within 10 days of imbibition, compared with <45% germination and 20% seedling emergence in freshly harvested seed. Hence, 3 months of after‐ripening could be considered a suitable treatment period for rice dormancy release. Dormancy release by after‐ripening is mainly correlated with a rapid decline in ABA content and increase in IAA content during imbibition. Subsequently, GA₁/ABA, GA₇/ABA, GA₁₂/ABA, GA₂₀/ABA and IAA/ABA ratios significantly increased, while GA₃/ABA, GA₄/ABA and GAs/IAA ratio significantly decreased in imbibed seeds following 3 months of after‐ripening, thereby altering α‐amylase activity during seed germination. Peak α‐amylase activity occurred at an earlier germination stage in after‐ripened seeds than in freshly harvested seeds. Expression of ABA, GA and IAA metabolism genes and dormancy‐related genes was regulated by after‐ripening time upon imbibition. Expression of OsCYP707A5, OsGA2ox1, OsGA2ox2, OsGA2ox3, OsILR1, OsGH3‐2, qLTG3‐1 and OsVP1 increased, while expression of Sdr4 decreased in imbibed seeds following 3 months of after‐ripening. Dormancy release through after‐ripening might be involved in weakening tissues covering the embryo via qLTG3‐1 and decreased ABA signalling and sensitivity via Sdr4 and OsVP1.     

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.     

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.     

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.     

Protein Patterns, Characterized by Computer Image Analysis, of Lentil Embryo Axes Germinating Under Salt Stress

Following 16, 40 and 64 h exposure to 0.33 M NaCl given after 8 h water imbibition, lentil seeds showed a gradual decrease of germination upon their transfer to water. These salt related changes were accompanied by modifications in the protein patterns of embryo axes as revealed by two-dimensional electrophoresis separation and by the computer image analysis of protein spots. In comparison with 8 h water imbibed seeds, prominent proteins comprised between the 5.1 – 7.6 pH isoelectric point in the first dimension and 75 – 50 kDa molecular mass in the second dimension showed a significant increase in their abundance as salt exposure increased. On transfer to water to complete germination, the content of many of these proteins decreased at 24h in 2 – 3 cm length embryo axes in comparison with the corresponding embryo axes of seeds continuously imbibed in water for 24 h. Some groups of proteins ranging between 15.5 – 17.3 kDa, already present after 8 h water imbibition, were not detectable after 24 h but were expressed in seeds exposed to NaCl and transferred to water for 24 h. Up- and down-regulated proteins in lentil embryo axes, imbibed under non-lethal salt stress conditions, have been tentatively identified by comparison with the protein map of germinating seeds of the model plant Arabidopsis. This revised version was published online in July 2006 with corrections to the Cover Date.     

ATP Synthesis during Water Imbibition in Caryopses of Genetically Dormant and Non-dormant Lines of Wild Oat (Avena fatua L.)

Levels of ATP in dry caryopses of wild oats (Avena fatua L.)were much lower than in imbibed seeds of the seven geneticallypure lines surveyed. The ATP content of the lines with highgenetic dormancy was consistently lower than the ATP contentof genetically non-dormant lines, but no significant correlationwith depth of dormancy was found apart from this. Massive increasesin ATP content occurred within 30 min of water uptake by caryopsesof both dormant and non-dormant lines. The synthetic pathwaystudied utilized inorganic phosphate with great avidity to formATP. The ability to form ATP upon imbibition was present inboth embryo and de-embryonated caryopsis. The ATP levels attainedin imbibing caryopses appeared sufficient to support considerablesynthetic activity, and this reduced the possibility that adeficiency in ATP was responsible for the maintenance of dormancyin such imbibed seeds. The low levels of inorganic phosphatein the embryos of genetically dormant lines of wild oat couldrepresent a limiting factor, if the active formation of ATPupon water imbibition resulted in a scarcity of phosphate forother reactions essential to germination. Key words: Avena fatua, ATP synthesis, Inorganic phosphorus, Seed dormancy, Germination, Water uptake