Seed coats: Structure,development, composition,and biotechnology

Summary Although seeds have been the subject of extensive studies for many years, their seed coats are just beginning to be examined from the perspective of molecular genetics and control of development. The seed coat, plays a vital role in the life cycle of plants by controlling the development of the embryo and determining seed dormancy and germination. Within the seed coat are a number of unique tissues that undergo differentiation to serve specific functions in the seed. A large number of genes are known to be specifically expressed within the seed coat tissues; however, very few of them are understood functionally. The seed coat synthesizes a wide range of novel compounds that may serve the plant in diverse ways, including defense and control of development. Many of the compounds are sources of industrial products and are components of food and feeds. The use of seed coat biotechnology to enhance seed quality and yield, or to generate novel components has not been exploited, largely because of lack of knowledge of the genetic systems that govern seed coat development and composition. In this review, we will examine the recent advances in seed coat, biology from the perspective of structure, composition and molecular genetics. We will consider the diverse avenues that are possible for seed coat biotechnology in the future. This review will focus principally on the seed coats of the Brassicaceae and Fabaceae as they allow us to merge the areas of molecular biology, physiology and structure to gain a perspective on the possibilities for seed coat modifications in the future. The authors have contributed equally and are considered first authors.     

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。     

Embryological features of Alhagi persarum (Fabaceae): Adapt to environmental constraints

Reproductive organs, in flowering plants, are sensitive to stressful environments. Alhagi persarum Boiss. & Buhse copes with the stresses and produce reproductive organs under difficult climatic conditions. Embryological characters of this plant were investigated for the first time using different microscopy and staining techniques. The results of this study showed unique reproductive characters and strategies in A. persarum that we named reproductive adaptation. These characters have roles in protection and nutrition of reproductive organs, some of which were visible in ovule: accumulation of phenolic compounds, presence of ovular endothelium with its cuticle coat, hypostase, postament, endosperm haustorium, presence of operculum, curvature of the embryonic axis. The other characters in the seed are macrosclereid cells with cuticle coat, double palisade layer and lignified tracheids in hilar groove. Thickness increasing of endothecium and exine are the adaptive characters in anther. Unlike many of the stress-sensitive plants, all developmental stages of the embryo sac, anther, pollen and pollen tube are without any defects in these stress-tolerant plants. Seed germination rate is low in this species that is due to the hardness of seed coat which causes seed deep exogenous dormancy. This dormancy is also a developmental program for stress tolerance to keep seed viability for a long time in difficult conditions.     

The role of the persistent fruit wall in seed water regulation in Raphanus raphanistrum (Brassicaceae)

Background and Aims

Dry fruits remain around the seeds at dispersal in a number of species, especially the Brassicaceae. Explanations for this vary, but usually involve mechanisms of innate dormancy. We speculate that, instead, a persistent fruit may give additional protection through control of dehydration, to species growing in arid or Mediterranean environments where water is sporadic.

Methods

X-rays and weight measurements were used to determine the extent to which Raphanus raphanistrum seeds within mature fruits imbibe water, and germination tests determined the roles of the fruit and seed coat in seed dormancy. Rates of water uptake and desiccation, and seedling emergence were compared with and without the fruit. Finally, germinability of seeds extracted from fruits was determined after various periods of moist conditions followed by a range of dry conditions.

Key Results

Most seeds rapidly take up water within the fruit, but they do not fully imbibe when compared with naked seeds. The seed coat is more important than the dry fruit wall in maintaining seed dormancy. The presence of a dry fruit slows emergence from the soil by up to 6–8 weeks. The fruit slows the rate of desiccation of the seed to a limited extent. The presence of the fruit for a few days during imbibition somehow primes more seeds to germinate than if the fruit is absent; longer moist periods within the pod appear to induce dormancy.

Conclusions

The fruit certainly modifies the seed environment as external conditions change between wet and dry, but not to a great extent. The major role seems to be: (a) the physical restriction of imbibition and germination; and (b) the release and then re-imposition of dormancy within the seed. The ecological significance of the results requires more research under field conditions.     

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.     

曼陀罗种子休眠机理与破眠方法研究

通过对曼陀罗种子生活力测定、发芽试验、吸水率测定及种子萌发抑制物研究,揭示曼陀罗种子休眠机理,并利用物理、化学法处理曼陀罗种子,以探寻打破曼陀罗种子休眠的最佳方法.结果表明:(1)新采收的曼陀罗种子为综合休眠,休眠原因包括:种皮障碍、缺少萌发所需激素以及种皮和种仁中存在萌发抑制物,其中种皮障碍是限制种子萌发的首要因素.(2)室温存储6个月可解除曼陀罗种子种仁的休眠,但种皮障碍始终是其种子萌发的限制因素.(3)机械摩擦、浓H2SO4处理和NaOH处理均可打破除曼陀罗种皮的休眠障碍,促进种子萌发,其中用10% NaOH处理90 min为破除曼陀罗种皮休眠障碍的最佳方法,且发芽率比对照提高了83%.     

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.     

种子物理休眠研究进展

种子物理休眠是由种皮不透水层引起的一种休眠类型,是植物在长期系统发育进程中获得的一种适应环境变化的特性。该文简述了种子物理休眠的定义与概念;从不透水层、种皮的特殊水孔器结构以及胚的形态特异性等方面,综述了物理休眠种子的形态特征、物理休眠与综合休眠的解除方法以及物理休眠的可能解除机制;利用Angiosperm Phylogeny Group Ⅲ(APG Ⅲ)系统分析了种子物理休眠的植物在系统发育中的位置;最后提出了今后种子物理休眠有待研究的主要问题。     

Ethylene in seed dormancy and germination

The role of ethylene in the release of primary and secondary dormancy and the germination of non-dormant seeds under normal and stressed conditions is considered. In many species, exogenous ethylene, or ethephon – an ethylene-releasing compound - stimulates seed germination that may be inhibited because of embryo or coat dormancy, adverse environmental conditions or inhibitors (e.g. abscisic acid, jasmonate). Ethylene can either act alone, or synergistically or additively with other factors. The immediate precursor of ethylene biosynthesis, 1-aminocyclopropane-1-carboxylic acid (ACC), may also improve seed germination, but usually less effectively. Dormant or non-dormant inhibited seeds have a lower ethylene production ability, and ACC and ACC oxidase activity than non-dormant, uninhibited seeds. Aminoethoxyvinyl-glycine (AVG) partially or markedly inhibits ethylene biosynthesis in dormant or non-dormant seeds, but does not affect seed germination. Ethylene binding is required in seeds of many species for dormancy release or germination under optimal or adverse conditions. There are examples where induction of seed germination by some stimulators requires ethylene action. However, the mechanism of ethylene action is almost unknown.
The evidence presented here shows that ethylene performs a relatively vital role in dormancy release and seed germination of most plant species studied.     

Genetic differences in seed longevity of various Arabidopsis mutants

Seeds gradually lose their viability during dry storage. The damage that occurs at the biochemical level can alter the seed physiological status and is affected by the storage conditions of the seeds. Although these environmental conditions controlling loss of viability have been investigated frequently, little information is available on the genetics of seed longevity. Using Arabidopsis mutants in defined developmental or biochemical pathways such as those affected in seed coat composition, seed dormancy, hormone function and control of oxidative stress, we tried to gain insight into the genes and mechanisms controlling viability of stored seeds. Mutations like abscisic acid insensitive3 ( abi3 ) as well as abscisic acid deficient1 ( aba1 ) show reduced longevity, which may be partially related to the seed dormancy phenotype of these mutants. Mutants with seed coat alterations, especially aberrant tests shape ( ats ), showed a stronger reduction in germination percentage after storage, indicating the importance of a 'functional' seed coat for seed longevity. A specific emphasis was placed on mutants affected in dealing with Reactive Oxygen Species (ROS). Because several pathways are involved in protection against ROS and because gene redundancy is a common feature in Arabidopsis , 'double' mutants were generated. These 'double' mutants and the corresponding single mutants were subjected to a controlled deterioration test (CDT) and a germination assay on hydrogen peroxide (H₂O₂) after prolonged storage at two relative humidities. CDT and germination on H₂O₂ affected all genotypes, although it appears that other effects like genetic background are more important than the deficiencies in the ROS scavenging pathway. Explanations for this limited effect of mutations affecting ROS scavenging are discussed.     

Does fire drive fatty acid composition in seed coats of physically dormant species?

• Seed dormancy is the key driver regulating seed germination, hence is fundamental to the seedling recruitment life-history stage and population persistence. However, despite the importance of physical dormancy (PY) in timing post-fire germination, the mechanism driving dormancy-break within seed coats remains surprisingly unclear. We suggest that seed coat chemistry may play an important role in controlling dormancy in species with PY. In particular, seed coat fatty acids (FAs) are hydrophobic, and have melting points within the range of seed dormancy-breaking temperatures. Furthermore, melting points of saturated FAs increase with increasing carbon chain length. We investigated whether fire could influence seed coat FA profiles and discuss their potential influence on dormancy mechanisms.
• Seed coat FAs of 25 species within the Faboideae, from fire-prone and fire-free ecosystems, were identified and quantified through GC–MS. Fatty acid profiles were interpreted in the context of species habitat and interspecific variation.
• Fatty acid compositions were distinct between species from fire-prone and fire-free habitats. Fire-prone species tended to have longer saturated FA chains, a lower ratio of saturated to unsaturated FA, and a slightly higher relative amount of FAs compared to fire-free species.
• The specific FA composition of seed coats of fire-prone species indicated a potential role of FAs in dormancy mechanisms. Overall, the distinct FA composition between fire-prone and fire-free species suggests that chemistry of the seed coat may be under selection pressure in fire-prone ecosystems.

     

Induction of secondary dormancy in Amaranthus caudatus seeds

Nondormant A. caudatus seeds germinated in the darkat temperatures between 20 and 35° but not at 45 °C.Incubation at this temperature for at least 10 h inhibited seedgermination over the temperature range 20 to 35 °C,temperatures previously suitable for germination. Thus incubation at 45°C induced secondary dormancy. Mechanical or chemicalscarification or exposure to pure oxygen caused complete or almost completegermination of dormant seeds although more slowly in comparison to nondormantseeds. Secondary dormant scarified seeds required a lower concentration of ABAthan nondormant seeds to inhibit germination. The high temperature, whichinduced dormancy, 45 °C, caused the seed coat to be partiallyresponsible for secondary dormancy. Involvement of ABA (synthesis orsensitivity) in the induction and/or maintenance of this dormancy should beconsidered.     

A major QTL controlling seed dormancy and pre-harvest sprouting resistance on chromosome 4A in a Chinese wheat landrace

Wheat pre-harvest sprouting (PHS) can cause significant reduction in yield and end-use quality of wheat grains in many wheat-growing areas worldwide. To identify a quantitative trait locus (QTL) for PHS resistance in wheat, seed dormancy and sprouting of matured spikes were investigated in a population of 162 recombinant inbred lines (RILs) derived from a cross between the white PHS-resistant Chinese landrace Totoumai A and the white PHS-susceptible cultivar Siyang 936. Following screening of 1,125 SSR primers, 236 were found to be polymorphic between parents, and were used to screen the mapping population. Both seed dormancy and PHS of matured spikes were evaluated by the percentage of germinated kernels under controlled moist conditions. Twelve SSR markers associated with both PHS and seed dormancy were located on the long arm of chromosome 4A. One QTL for both seed dormancy and PHS resistance was detected on chromosome 4AL. Two SSR markers, Xbarc 170 and Xgwm 397, are 9.14 cM apart, and flanked the QTL that explained 28.3% of the phenotypic variation for seed dormancy and 30.6% for PHS resistance. This QTL most likely contributed to both long seed dormancy period and enhanced PHS resistance. Therefore, this QTL is most likely responsible for both seed dormancy and PHS resistance. The SSR markers linked to the QTL can be used for marker-assisted selection of PHS-resistant white wheat cultivars. Shi-Bin Cai and Cui-Xia Chen contributed equally to this work.     

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.     

Seed coat development, anatomy and scanning electron microscopy of Harpagophytum procumbens (Devil's Claw), Pedaliaceae

Seed coat development of Harpagophytum procumbens (Devil's Claw) and the possible role of the mature seed coat in seed dormancy were studied by light microscopy (LM), transmission electron microscopy (TEM) and environmental scanning electron microscopy (ESEM). Very young ovules of H. procumbens have a single thick integument consisting of densely packed thin-walled parenchyma cells that are uniform in shape and size. During later developmental stages the parenchyma cells differentiate into 4 different zones. Zone 1 is the multi-layered inner epidermis of the single integument that eventually develops into a tough impenetrable covering that tightly encloses the embryo. The inner epidermis is delineated on the inside by a few layers of collapsed remnant endosperm cell wall layers and on the outside by remnant cell wall layers of zone 2, also called the middle layer. Together with the inner epidermis these remnant cell wall layers from collapsed cells may contribute towards seed coat impermeability. Zone 2 underneath the inner epidermis consists of large thin-walled parenchyma cells. Zone 3 is the sub-epidermal layers underneath the outer epidermis referred to as a hypodermis and zone 4 is the single outer seed coat epidermal layer. Both zones 3 and 4 develop unusual secondary wall thickenings. The primary cell walls of the outer epidermis and hypodermis disintegrated during the final stages of seed maturation, leaving only a scaffold of these secondary cell wall thickenings. In the mature seed coat the outer fibrillar seed coat consists of the outer epidermis and hypodermis and separates easily to reveal the dense, smooth inner epidermis of the seed coat. Outer epidermal and hypodermal wall thickenings develop over primary pit fields and arise from the deposition of secondary cell wall material in the form of alternative electron dense and electron lucent layers. ESEM studies showed that the outer epidermal and hypodermal seed coat layers are exceptionally hygroscopic. At 100% relative humidity within the ESEM chamber, drops of water readily condense on the seed surface and react in various ways with the seed coat components, resulting in the swelling and expansion of the wall thickenings. The flexible fibrous outer seed coat epidermis and hypodermis may enhance soil seed contact and retention of water, while the inner seed coat epidermis maintains structural and perhaps chemical seed dormancy due to the possible presence of inhibitors.     

Changes in phenolic acids and abscisic acid in sugar pine seed coats during stratification

The phenolic acids and abscisic acid (ABA) of sugar pine ( Pinus lambertiana Dougl.) seeds coats, separated by high-pressure liquid chromatography, were analyzed during 90 days stratification of the seeds. Although levels of seed coat phenolic acids and ABA declined significantly during, stratification, this decrease did not appear to be responsible for the loss of dormancy due to stratification. Lack of improved germination following washing, cracking, or removal of the seed coats, plus additional evidence, did not support a significant role for the seed coat in the dormancy of sugar pine seeds.     

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。     

More than just a coating: Ecological importance,taxonomic occurrence and phylogenetic relationships of seed coat mucilage

Studies on the ecological importance of seed coat mucilage have provided valuable information about its roles in critical stages of the plant life cycle. Seed mucilage may, by providing a moist environment and maintaining metabolic activity in the seed, promote seed development. In seed dispersal, seed mucilage influences topochory, epizoochory, endozoochory and hydrochory by anchorage of seeds to soil surface, lubrication or changing the specific weight of the seed. In arid environments, seed mucilage can prevent seeds from drying or initiate DNA repair mechanisms, thereby maintaining the soil seed bank. Seed mucilage reduces oxygen diffusion to the seed and thus has a role in regulating seed dormancy. Due to it being hydrophilous, acting as a physical barrier and containing chemicals, seed mucilage is proposed to promote seed germination in favorable environments. In seedling growth, seed mucilage may lubricate the radicle as it penetrates the soil and be degraded by soil microfloras and thus promote seedling growth. Further investigation of seed mucilage for more species in diverse habitats from the perspectives of evolution, genetics, proteomics, phylogeny and plant–microbe interactions would contribute substantially to our understanding about its ecological importance.     

Changes in dormancy levels ofFraxinus excelsior L. embryos at different stages of morphological and physiological maturity

The dormancy status ofFraxinus excelsior embryos at different developmental stages under environmental conditions was examined over a period of 2 years. For each sampling date the length of the fruit, of the seed, and of the embryo were measured, and the embryological stage determined. The depth of dormancy was assayed by the germination behaviour of isolated embryos under aseptic conditions on an agar medium without nutrients. As an approach towards a quantitative estimate of the dormancy status, the degree of inhibiton of germinative growth in the embryonic organs was evaluated on the basis of four categories from none to full germinative growth. From these ratings a dormancy index was calculated, expressing the mean dormancy status of the embryos at a given date. Embryo dormancy already became apparent during embryogenesis and reached its highest level during the later phase of reserve deposition in the seed. A marked loss of embryo dormancy occurred during the phase of maturation drying in autumn, followed by a moderate increase in winter. In hydrated seeds in spring the embryo was gradually released from dormancy and enlarged further. In maintaining the embryo ofF. excelsior in a developmental but not germinative mode, dormancy mechanisms within the embryo and the endosperm, combined with environmental factors, may be involved.