Changes in Catechol Oxidase and Permeability to Water in Seed Coats of Pisum elatius during Seed Development and Maturation

In the developing seed coat of Pisum elatius, o-dihydroxyphenols are present in appreciable amounts at all stages of development. However, catechol oxidase activity rises sharply during the later stages of development, shows a further abrupt rise during dehydration of the seed coat, and then decreases. It is suggested that a tanning reaction is induced by the contact of enzyme with its substrate while cell membranes are ruptured, and that this reaction renders the seed coats impermeable. The entire chain of events does not occur in Pisum sativum which has permeable seed coats.     

Relation Between the Anatomy of the Testa, Water Permeability and the Presence of Phenolics in the Genus Pisum

The seed coats of Pisum elatius, P. fulvum and P. humile areimpermeable to water while those of P. sativum and P. humilex P. sativum are permeable. The anatomical structure of theseed coats and the location of phenolics and quinones in thecells is described. The barriers to permeation of water in theimpermeable seeds are the continuous, very hard, pectinaceouslayer of the caps of the palisade cells and the presence ofquinones in either the palisade or osteosclereid cells, in acontinuous layer of these cells. In water permeable seeds thecaps are looser and quinones discontinuous or absent in palisadeor osteosclereid cells. Pisum, testa, water permeability, quinones, phenolics, palisade cells, osteosclereids, pectinaceous caps     

Peroxidase involvement in lignification in water-impermeable seed coats of weedy leguminous and malvaceous species

Abstract. The development of water impermeable seed coats of two members each of the leguminoseae family [ Crotalaria spectabilis Roth, Sesbania exaltata (Raf) Cory] and the malvaceae family [Anoda cristata (L.) Schlecht, Abutilon theophrasti Medic.] was investigated. Highest peroxidase (POD) activity of Anoda and Abutilon seed coat extracts was highly correlated with the developmental stages when soluble phenolics were maximally converted into lignin. Although extensive lignification occurred during seed coat development in both legumes, the patterns of POD activity, soluble phenolic levels and time of lignification were different from those of the malvaceous species. POD activity levels in developing coats of the malvaceous seeds increased as phenolics decreased. Both POD activity and phenolic levels decreased during seed coat development of the legumes. POD was immunocytochemically and immunochemically detected in seed coats of all four species; however, results for polyphenol oxidase were negative. The results confirmed POD involvement in lignification of leguminous and malvaecous species and support and extend our earlier view that POD is involved in lignin formation during development of impermeable seed coats.     

Relationship between Colour, Phenolic Content and Impermeability in the Seed Coat of various Trifolium subterraneum L. genotypes

The relationship between seed colour, phenol content of thetesta and water impermeability in dark and light seeded genotypesof Trifolium subterraneum L. was investigated. The developmentof water impermeability and catechol oxidase activity in expandingseeds of two genotypes was monitored. The results show catecholoxidase activity decreases as seed colour changes from greento purple, but the potential to become impermeable with dehydrationis not acquired until later in seed development. Both waterpermeable and impermeable seeds of dark coloured genotypes containsubstantial amounts of phenol in the lumen of the Malpighiancells; light coloured testas do not. It is concluded that the darkening of the testa of Trifoliumsubterraneum is associated with oxidation of phenol by catecholoxidase but that development of impermeability is independentof this process. Trifolium subterraneum L., subterranean clover, seed coat impermeability, catechol oxidase, phenol, testa colour     

Patterns and kinetics of water uptake by soybean seeds

Soybean [Glycine max (L.) Merr.] plants produce some seeds (called stone or impermeable seeds) that do not take up water for long periods of time. The present investigation confirmed that the stone seed trait is a feature of the seed coat: isolated embryos from both stone and permeable seeds took up water equally quickly. A whole, permeable seed typically imbibed water initially through its dorsal side, forming wrinkles in the seed coat and delivering water to the underlying cotyledons. Later, some lateral movement of water through the coat occurred, presumably through the air spaces of the osteosclereid layer. Imbibition by seeds was a two-phase process, the first dominated by hydration of the seed coat and the second by hydration of the cotyledons, which was rate-limited by the coat. When hydrated, coats of stone seeds were permeable to water but their hydraulic conductivity, as measured with a pressure probe, was smaller than that of coats from permeable seeds by a factor of five. Hydrated coats of both permeable and stone seeds showed weak osmometer properties.     

Cracks in the palisade cuticle of soybean seed coats correlate with their permeability to water

BACKGROUND AND AIMS: Soybean (Glycine max) is among the many legumes that are well known for 'hardseededness'. This feature can be beneficial for long-term seed survival, but is undesirable for the food processing industry. There is substantial disagreement concerning the mechanisms and related structures that control the permeability properties of soybean seed coats. In this work, the structural component that controls water entry into the seed is identified. METHODS: Six soybean cultivars were tested for their seed coat permeabilities to water. To identify the structural feature(s) that may contribute to the determination of these permeabilities, fluorescent tracer dyes, and light and electron microscopic techniques were used. KEY RESULTS: The cultivar 'Tachanagaha' has the most permeable seed coat, 'OX 951' the least permeable seed coat, and the permeabilities of the rest ('Harovinton', 'Williams', 'Clark L 67-3469', and 'Harosoy 63') are intermediate. All seeds have surface deposits, depressions, a light line, and a cuticle about 0.2 microm thick overlaying the palisade layer. In permeable cultivars the cuticle tends to break, whereas in impermeable seeds of 'OX 951' it remains intact. In the case of permeable seed coats, the majority of the cracks are from 1 to 5 micro m wide and from 20 to 200 micro m long, and occur more frequently on the dorsal side than in other regions of the seed coat, a position that correlates with the site of initial water uptake. CONCLUSIONS: The cuticle of the palisade layer is the key factor that determines the permeability property of a soybean seed coat. The cuticle of a permeable seed coat is mechanically weak and develops small cracks through which water can pass. The cuticle of an impermeable seed coat is mechanically strong and does not crack under normal circumstances.     

Control of seed coat thickness and permeability in soybean : a possible adaptation to stress

Although the seed coat, through its thickness and permeability, often regulates seed germination, very little is known about the control of its development. Using soybean (Glycine max [L.] Merrill) explants, podbearing cuttings in which defined solutions can be substituted for the roots, we have demonstrated that cytokinin and mineral nutrients moving through the xylem can control soybean seed coat development. Lack of cytokinin and minerals in the culture solution, causes a thicker, less permeable seed coat to develop. The seeds with thickened coats will imbibe water rapidly if scarified; furthermore, these scratched seeds also germinate and produce normal plants. Inasmuch as stress (e.g. drought) decreases mineral assimilation and cytokinin production by the roots, the resulting delay in germination could be an adaptive response to stress.     

Development of seed coat-imposed dormancy during seed maturation in Cynoglossum officinale

The relationship between seed phenolics and appearance of seed coat–imposed dormancy during seed development in Cynoglossum officinale L. was studied. Up to 24 days after anthesis, seeds failed to germinate upon imbibition in Petri dishes at 25°C. At 44 days after anthesis, seeds were fully germinable; removal of seed coats did not improve their germination or O₂ uptake. At 72 days after anthesis, mature seeds at the base of the cyme did not germinate unless their coats were removed. Removal of seed coat also stimulated O₂ uptake at this harvest date. The methanol-soluble phenolic content of the seeds increased during the early stages of seed development, in both the seed coat and the embryo. As seed development continued, the methanol-soluble phenolic content of the embryo stabilized, but that of the seed coat declined. This decline was associated with an increase in the thioglycolic acid–soluble phenolics, presumably lignins, in the seed coat. These results suggest that polymerization of methanol–soluble phenolics into lignins in the seed coat during later stages of seed development renders the seed coat of C. officinale impermeable to 03, and thus keeps the seed dormant.     

Seed coat composition in black and white soybean seeds with differential water permeability

• The seed coat composition of white (JS 335) and black (Bhatt) soybean (Glycine max (L.) Merr) having different water permeability was studied.
• Phenols, tannins and proteins were measured, as well as trace elements and metabolites in the seed coats.
• The seed coat of Bhatt was impermeable and imposed dormancy, while that of JS 335 was permeable and seeds exhibited imbibitional injury. Bhatt seed coats contained comparatively higher concentrations of phenols, tannins, proteins, Fe and Cu than those of JS 335. Metabolites of seed coats of both genotypes contained 164 compounds, among which only 14 were common to both cultivars, while the remaining 79 and 71 compounds were unique to JS 331 and Bhatt, respectively.
• Phenols are the main compounds responsible for seed coat impermeability and accumulate in palisade cells of Bhatt, providing impermeability and strength to the seed coat. JS 335 had more cracked seed coats, mainly due to their lower tannin content. Alkanes, esters, carboxylic acids and alcohols were common to both genotypes, while cyclic thiocarbamate (1.07%), monoterpene alcohols (1.07%), nitric esters (1.07%), phenoxazine (1.07%) and sulphoxide (1.07%) compounds were unique to the JS 335 seed coat, while aldehydes (2.35%), amides (1.17%), azoles (1.17%) and sugar moieties (1.17%) were unique to Bhatt seed coats. This study provides a platform for isolation and understanding of each identified compound for its function in seed coat permeability.

     

Seed development in the rare cold desert sand dune shrub Eremosparton songoricum and a comparison with other papilionoid legumes

No study has yet been carried out on seed development in a cold desert sand dune papilionoid legume. Thus, our primary aims were to (i) monitor seed development in the cold desert sand dune species Eremosparton songoricum from the time of pollination to seed maturity, and (ii) compare seed development in this species with that in other species of papilionoid legumes. Fruit and seed size, mass and seed moisture content, and seed imbibition, germination, desiccation tolerance and water retention during development (pollination to seed maturity) were monitored in the papilionaceous shrub E. songoricum in the Gurbantunggut Desert of northwest China. The duration of seed development was 40 days. Seeds reached physiological maturity 28 days after pollination (DAP), at which time 58% of them germinated and they had developed desiccation tolerance. Seeds became impermeable 36–40 DAP, when their moisture content was about 10%. The final stage of maturation drying occurred via loss of water through the hilum. The developmental stages and their timing (DAP) in seeds of E. songoricum are generally similar to those reported for other papilionaceous legumes with a water‐impermeable seed coat (physical dormancy). In general, the developmental features of seeds with water‐impermeable coats at maturity do not appear to be specific to habitat or phylogeny.     

Identification of a single gene for seed coat impermeability in soybean PI 594619

Key message

Inheritance studies and molecular mapping identified a single dominant gene that conditions seed coat impermeability in soybean PI 594619.

Abstract

High temperatures during seed fill increase the occurrence of soybeans with impermeable seed coat, which is associated with non-uniform and delayed germination and emergence. This can be an issue in soybean production areas with excessively high-temperature environments. The objectives of the present study were to investigate the inheritance of impermeable seed coat under a high-temperature environment in the midsouthern United States and to map the gene(s) that affect this trait in a germplasm line with impermeable seed coat (PI 594619). Crosses were made between PI 594619 and an accession with permeable seed coat at Stoneville, MS in 2008. The parental lines and the segregating populations from reciprocal crosses were grown in Stoneville in 2009. Ninety-nine F_2:3 families and parents were also grown at Stoneville, MS in 2011. Seeds were assayed for percent impermeable seed coat using the standard germination test. Genetic analysis of the F₂ populations and F_2:3 families indicated that seed coat impermeability in PI 594619 is controlled by a single major gene, with impermeable seed coat being dominant to permeable seed coat. Molecular mapping positioned this gene on CHR 2 between markers Sat_202 and Satt459. The designation of Isc (impermeable seed coat) for this single gene has been approved by the Soybean Genetics Committee. Selection of the recessive form (isc) may be important in developing cultivars with permeable seed coat for high-heat production environments. The single-gene nature of impermeable seed coat may also have potential for being utilized in reducing seed damage caused by weathering and mold.     

A Single-Nucleotide Polymorphism in an Endo-1,4-β-Glucanase Gene Controls Seed Coat Permeability in Soybean

Physical dormancy, a structural feature of the seed coat known as hard seededness, is an important characteristic for adaptation of plants against unstable and unpredictable environments. To dissect the molecular basis of qHS1, a quantitative trait locus for hard seededness in soybean (Glycine max (L) Merr.), we developed a near-isogenic line (NIL) of a permeable (soft-seeded) cultivar, Tachinagaha, containing a hard-seed allele from wild soybean (G. soja) introduced by successive backcrossings. The hard-seed allele made the seed coat of Tachinagaha more rigid by increasing the amount of β-1,4-glucans in the outer layer of palisade cells of the seed coat on the dorsal side of seeds, known to be a point of entrance of water. Fine-mapping and subsequent expression and sequencing analyses revealed that qHS1 encodes an endo-1,4-β-glucanase. A single-nucleotide polymorphism (SNP) introduced an amino acid substitution in a substrate-binding cleft of the enzyme, possibly reducing or eliminating its affinity for substrates in permeable cultivars. Introduction of the genomic region of qHS1 from the impermeable (hard-seeded) NIL into the permeable cultivar Kariyutaka resulted in accumulation of β-1,4-glucan in the outer layer of palisade cells and production of hard seeds. The SNP allele found in the NIL was further associated with the occurrence of hard seeds in soybean cultivars of various origins. The findings of this and previous studies may indicate that qHS1 is involved in the accumulation of β-1,4-glucan derivatives such as xyloglucan and/or β-(1,3)(1,4)-glucan that reinforce the impermeability of seed coats in soybean.     

Crypsis hypothesis as an explanation for evolution of impermeable coats in seeds is anecdotal

Impermeable seed/fruit coat, i.e. physical dormancy (PY) occurring in seeds of many genera of 19 angiosperm plant families has been traditionally viewed as a form of dormancy that regulates germination timing. However, this view was recently challenged by an alternative explanation claiming that the impermeable seed coat evolved as a coping mechanism to escape predators, i.e. crypsis hypothesis. Here, I wish to call for more careful attention on crypsis as an evolutionary factor because (1) information of volatile compounds is not known in PY families except Fabaceace; (2) impermeability is not induced until the moisture content of the seeds drops below species-specific threshold suggesting that drying determines development of impermeable seed coats; (3) the crypsis hypothesis does not explain the year-to-year or between site variations in proportions of impermeable seeds produced by plants; and (4) dry seeds of species from non-PY families also do not emit volatile compounds, and do not develop impermeable seed coats. For these reasons, it appears crypsis might be an exaptation, i.e., a trait that performs a function for which it was not originally evolved. Based on the available evidence, it is suggested that climate drying might have resulted in evolution of impermeable seed coats.     

Imbibition and Germination: Influence of Hard Seed Coats on RNA Metabolism

RNA metabolism of Indian rice grass seeds was studied in relation to imbibition and germination. These seeds germinate only after scarification of the seed coats. The hard seed coats restrict germination but not water intake or changes in the quantity and quality of RNA formed during early hours of soaking. These changes differ markedly from those in scarified (H₂SO₄ treated) seeds which are able to germinate. Gibberellic acid hastens germination of scarified seeds and causes changes in the population of RNA transcribed.     

Water impermeable seed dormancy

Viable seeds that do not imbibe water and thus fail to germinate in an apparently favorable environment are commonly termed impermeable or hard seed. This physical, exogenous dormancy is especially common in species of the Fabaceae. The ecological significance of hard seed includes the ability to rapidly recolonize burnt areas after fire and to withstand ingestion by animals and birds. Advantages and problems that hard seed cause in agriculture are discussed. Species from different families with impermeable seeds appear to have in common a layer of macrosclerid cells that form a palisade layer in the testa. The term strophiole and its contradictory use in botanical literature are discussed. Genetic factors and environmental conditions both affect the proportion of impermeable seeds produced. Methods of artificially softening impermeable seeds include acid and solvent, soaking, mechanical scarification, pressure, percussion, freezing, heating, and radiation treatments that can result in a change in germination from less than 20% in some untreated species up to 90% or more in treated species. Natural softening involves high temperatures and temperature fluctuations and the degree of desiccation of the seed. The mechanism of water impermeability is related to the testa and is thought to involve waterproofing substances including wax, lignin, tannin, suberin, pectin, and quinone derivatives. The hilum acts as a hygroscopic valve that prevents water uptake but allows water loss to occur at low relative humidities in some species. The strophiole is an area of weakness in the testa of some Papilionoideae while the chalaza region has been determined as an area of weakness inPisum andGossypium. The water impermeable status of some species is reversible at a seed moisture content greater than 10%. The hard seed of a species can be described both in terms of the amount and the degree of impermeability.     

Seed dormancy of Cardiospermum halicacabum (Sapindaceae) from three precipitation zones in Sri Lanka

• This study investigated seed germination of Cardiospermum halicacabum, a medicinally important invasive species.
• We compared mass, moisture content (MC), dormancy and dormancy‐breaking treatments and imbibition and germination of scarified and non‐scarified seeds of C. halicacabum from a low‐elevation dry zone (DZ), low‐elevation wet zone (WZ1) and mid‐elevation wet zone (WZ2) in Sri Lanka to test the hypothesis that the percentage of seeds with water‐impermeable seed coats (physical dormancy, PY) decreases with increased precipitation.
• Seed mass was higher in WZ2 than in DZ and WZ1, while seed MC did not vary among the zones. All scarified DZ, WZ1 and WZ2 and non‐scarified DZ and WZ1 seeds imbibed water, but only a few non‐scarified WZ2 seeds did so. When DZ and WZ1 seeds were desiccated, MC and percentage imbibition decreased, showing that these seeds have the ability to develop PY. GA₃ promoted germination of embryos excised from fresh DZ and WZ1 seeds and of scarified WZ2 seeds.
• At maturity, seeds from DZ and WZ1 had only physiological dormancy (PD), while those from WZ2 had combinational dormancy (PY+PD). Thus, our hypothesis was not supported. Since a high percentage of excised embryos developed into normal seedlings; this is a low‐cost method to produce C. halicacabum plants for medicinal and ornamental purposes.

     

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     

Physical dormancy and histological features of seeds of five Acacia species (Fabaceae) from xerophytic forests in central Argentina

Physical dormancy (impermeability of seed coats to water) is related to histological features of the seed coat. This mechanism has ecological importance since it determines the time and space of germination. The aim of the present study was to compare the histology and impermeability of the seed coat in five Neotropical Acacia species from xerophytic forests of central Argentina: Acacia aroma, A. caven, A. atramentaria, A. gilliesii and A. praecox. An imbibition experiment was performed to determine the presence or absence of physical dormancy. Seed coat structure was studied through histochemical analysis. The seeds of A. gilliesii and A. praecox were treated with ammonium ferrous sulfate to identify the sites of water entry. Acacia aroma, A. caven and A. atramentaria exhibited physical dormancy; the seed coat was very thick and compact, with a wide, sclerified parenchyma and a “water gap” for water uptake. Seed coat impermeability in these species was mainly attributed to characteristics of the lignified epidermis. By contrast, A. gilliesii and A. praecox did not have physical dormancy and showed thin seed coats with a much narrower sclerified parenchyma. Water entered the seeds of A. gilliesii and A. praecox not only through the hilar zone but also through the entire surface of the seed coat. Differences in the seed coat structure among species could be related to different regenerative responses to environmental conditions that would facilitate the coexistence of these Acacia species in the xerophytic forests of Córdoba, Argentina.     

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.     

The Function of the Hilum in Some Papilionaceae in Relation to the Ripening of the Seed and the Permeability of the Testa

In seeds of Trifolium repens, T. pratense, and Lupinus arboreus,the hilum is a hygroscopically activated valve in the impermeableepidermis of the testa. When relative humidity was low the fissurein the hilum opened permitting the seed to dry out; when therelative humidity was high the fissure closed obstructing theabsorption of moisture. During seed-ripening the moisture contentfell readily to approximately 25 per cent., and thereafter moreslowly until the epidermis became impermeable at approximately14 per cent, moisture content. Further drying of the seed tookplace only by diffusion of water vapour through the hilum. ‘Hard’seeds tended to have a moisture content in equilibrium withthe lowest relative humidity to which they had been exposed.They absorbed moisture under conditions of gradually increasingrelative humidity such that the hilar fissure remained open.The duration of the impermeable condition increased with thedegree of desiccation brought about by loss of water throughthe hilum.