Generative ontogeny in Tiquilia (Ehretiaceae: Boraginales) and phylogenetic implications

Tiquilia is very different from the other members of the Ehretiaceae (Boraginales) in many aspects of morphology and ecology. Because detailed knowledge about flower and fruit traits is necessary to reliably infer character evolution of and within Tiquilia, we investigated flower to fruit ontogeny in eight species of Tiquilia using light and electron microscopy. Tiquilia accumulated a number of autapomorphies such as the prostrate growth form, the lack of lateral and ventral bundles in the gynoecium, and the formation of nutlet‐like mericarpids as dispersal units instead of more or less succulent drupes. The internal architecture of the superior bicarpellate ovary resulted from the development of several secondary septa including apical, basal and false septa, as it has been reported also from other Boraginales. However, no character found in Tiquilia can be regarded as synapomorphic with any other taxon of the Ehretiaceae. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 520–534.     

Morphology and anatomy of physical dormancy in Ipomoea lacunosa: identification of the water gap in seeds of Convolvulaceae (Solanales)

BACKGROUND AND AIMS: Convolvulaceae is the most advanced plant family (asterid clade) that produces seeds with physical dormancy (water-impermeable seed coat). There are several different opinions about the nature of the specialized structure ('water gap') in the seed coat through which water initially enters seeds of Convolvulaceae, but none of them has been documented clearly. The primary aim of the study was to identify the water gap in seeds of Ipomoea lacunosa (Convolvulaceae) and to describe its morphology, anatomy and function. METHODS: Light microscopy, scanning electron microscopy, tissue-sectioning, dye-tracking and blocking experiments were used to describe the morphology, anatomy and function of the water gap in seeds of I. lacunosa. KEY RESULTS: Dormancy-breaking treatments caused slits to form around the two bulges on the seed coat adjacent to the hilum, and dye entered the seed only via the disrupted bulges. Bulge anatomy differs from that of the rest of the seed coat. Sclereid cells of the bulges are more compacted and elongated than those in the hilum pad and in the rest of the seed coat away from the bulges. CONCLUSIONS: The transition area between elongated and square-shaped sclereid cells is the place where the water gap opens. Morphology/anatomy of the water gap in Convolvulaceae differs from that of taxa in the other 11 angiosperm plant families that produce seeds with physical dormancy for which it has been described.     

Imbibition of Swietenia macrophylla (Meliaceae) seeds: the role of stomata

BACKGROUND AND AIMS: The occurrence of stomata in seed coats is uncommon and there is limited information about their function(s). The aim of this study was to verify the distribution of stomata in seed coats of Swietenia macrophylla and to relate it to the imbibition process and aspects of the structure of the outer integument layers. METHODS: For the structural and ultrastructural studies, the seeds were processed using the usual techniques and studied under light and scanning electron microscopes. Histochemical tests were employed to identify the cell wall composition in the different seed coat portions. To assess the role of the stomata in the imbibition, non-impervious seeds were compared with partially impervious ones, in which only the embryo, median or hilar regions were left free. Further, the apoplastic pathway marker was employed to confirm the role of the stomata as sites of water passage during imbibition. KEY RESULTS: A positive relationship was observed between seed coat thickness and stomata density. The stomata were devoid of movement, with a large pore. They occurred in large numbers in the embryo region and extended with lower frequency towards the wing. Imbibition rates were related to stomata density, suggesting that the stomata act as preferential sites for water entry in the S. macrophylla seeds. CONCLUSIONS: At maturity, the stomata in the seed coat play a significant role in seed imbibition. The data may also infer that these permanently opened stomata have an important role in gas exchange during seed development, aiding embryo respiration.     

Seed removal,seed dispersers,and the allocation of tissues in Myrtaceae seeds

To avoid seed predation, plants may invest in protective seed tissues. Often related to seed size, allocation in seeds' physical defenses can also be influenced by dispersers. We explore the relationships between seed traits (seed mass and hardness) and seed removal in 22 Myrtaceae species of the Brazilian Atlantic Forest, a dominant and diverse fleshy-fruited taxon dispersed by birds, rodents, and other mammals. Our goal is to understand how seed traits influence seed removal rates, and whether dispersers can affect tissue allocation in the seed coat. Seeds were exposed to field removal experiments. In the laboratory, total seed mass and seed coat mass were obtained. To evaluate the influence of seed traits on removal, we performed Kruskal–Wallis and Simple Linear Regression tests. We assessed seed coat and seed mass covariation through standardized major-axis allometric regressions. Harder seeds were larger than softer ones. Seed traits affect removal rates, as tougher and heavier seeds had lower removal. Seed mass significantly predicts seed coat proportion in seven of the 14 species tested. Bird-dispersed species tend to exhibit lower proportions of seed coat as seed mass increases, whereas rodent-dispersed species apparently present the opposite trend, with seed coat proportion increasing with seed mass. Such difference may be caused by the contrasting seed predation pressure represented by birds and rodents. Energy allocation for defense, expressed in seed coat proportion, is greater in large seeds, as these are mostly dispersed by rodents whose propensity to cache and disperse seeds is greater for large and well-protected seeds.     

距药姜种子解剖学和组织化学研究

距药姜种子解剖学和组织化学研究表明,种子包括种皮、外胚乳、内胚乳和胚。外皮由1层表皮细胞构成,细胞壁纤维素质且明显增厚,中种皮可分为1层细胞的下皮层、半透明细胞层和2-4层细胞的色素细胞层,下皮层和色素细胞层的细胞内充满棕红色色素;内种皮由1层砖形薄壁细胞构成。珠孔区有珠孔领和孔盖的分化,但珠孔领分化不完善。合点区内种皮出现缺口,缺口间充满合点区色素细胞,其整体轮廓成新月形。外胚乳细胞壁平直,细胞内充满淀粉。内胚乳可分为多细胞区简细胞区两部分,内胚乳细胞界限不清,内含物主要是蛋白质,胚少有分化,含脂类、蛋白质、多糖,另外,还对姜花族的种子解剖学特征进行了初步的系统学分析。     

茴香砂仁种子的解剖学和组织化学研究

茴香砂仁种子的假种皮膜质,由内、外表皮及其间的数层薄壁细胞构成,种皮黑褐色,由外种皮、中种皮和内种皮组成,外种皮为1层表皮细胞;中种皮由1层细胞的下皮层,1层细胞的半透明细胞层、3-4层薄壁细胞的中种皮薄壁细胞层和1层细胞的色素细胞层组成,内种皮由1层径向延长的细胞构成,内切向壁与部分径向壁非常增厚,种子珠区分化出珠孔领,孔盖和珠孔区薄壁细胞,合点区内种皮出现缺口,缺口间的合点区色素细胞群整体轮廓呈刺叭状,珠孔端的则为1层细胞,细胞内含蛋白质、多糖、脂类物质,胚含量脂类物质,还含有蛋白质与多糖。     

Development of ovule and seed in Rapateaceae

VENTURELLI, M. & BOUMAN, F., 1988. Development of ovule and seed in Rapateaceae. The structure of the ovules and/or seeds of twelve species of Rapateaceae were studied, some additional embryological characters also being recorded. The ovules are always anatropous, bitegmic and crassinucellate, but they differ in the shape, size and in thickness of the outer integument. In Rapateaceae the outer integument is initiated subdermally. The seed coat of the Rapateaceae shows two mechanical layers: an endotesta with silica present as bodies or as incrustations in cell walls, in conjunction with an exotegmen with a jigsaw cell pattern complicated by a labyrinth-like sculpturing of the outer cell walls. The innermost layer of the inner integument is tanniniferous. Large hilar scars with tracheidal plates on the corresponding fruit wall and a persistent obturator are recorded in Rapateaceae. On the basis of embryological characters the family fits well into the Commelinales. Testa structure most closely resembles that of the Commelinaceae. The differences in ovule and seed structure agree with the currently accepted tribal classification.     

Strophiole of seeds of the black locust acts as a water gap

In Japan the black locust (Robinia pseudoacacia L.) is undergoing rapid habitat expansion, which has an adverse effect on native vegetation. It is therefore a priority to clarify the regeneration characteristics of the black locust and establish adequate management of this invasive species in Japan. To determine the germination characteristics of physically dormant black locust seeds, we observed anatomical features of the seed coat and identified the water gap that acts as a signal detector. Our microscopic observations showed that seed coats of this species had hilum, micropyle and strophiole. The anatomical features of these regions correspond to the general characteristics of papilionoid legume seeds. Based on our microscopic observations, water absorption blocking experiments and a dye tracking experiment, we identified the strophiole as a water gap in black locust seeds. Our results suggest that the opening of the strophiole is important for water uptake to the embryo and subsequent germination of black locust seeds under natural conditions.     

Phosphoenolpyruvate carboxykinase in developing pea seeds is associated with tissues involved in solute transport and is nitrogen-responsive

The aim of this work was to investigate the occurrence of phosphoenolpyruvate carboxykinase (PEPCK) in developing pea (Pisum sativum) seeds in relation to their nitrogen supply. PEPCK was present throughout development, with the peak of PEPCK protein and activity in the seed coat and cotyledons preceding protein accumulation in the cotyledons. It showed a different developmental pattern from enzymes involved in amino acid metabolism (phosphoenolpyruvate carboxylase, glutamine synthetase and glutamate dehydrogenase). Immunolocalization showed that PEPCK was present in parts of the developing seed that are involved in the transport and metabolism of assimilates. Early in development, it was associated with the inner integument of the ovule, the endospermic cytoplasm and the outer cells of the embryo. In the middle of development, around the peak of activity, PEPCK was abundant at the outer surface of the developing cotyledons, in the embryonic axis and in the vasculature of the seed coat. Later in development, PEPCK was associated with the embryonic leaf primordia and meristem and cortex of the radicle. PEPCK protein was strongly induced in vitro in the seed coat by nitrate, ammonium and asparagine, in the cotyledons by asparagine and in planta by the supply of nitrogen, which led to an increase in asparagine secretion by empty seed coats. It is suggested that PEPCK is involved in the metabolism of nitrogenous solutes in developing pea seeds.     

The role of the seed coat in adaptation of dimorphic seeds of the euhalophyte Suaeda salsa to salinity

The role of the seed coat in adaptation of dimorphic seeds of the euhalophyte Suaeda salsa to salinity was investigated during germination and early seedling growth. Black and brown seeds were treated with chloroform for 1 min before the extract was used to analyze waxes and the seeds to investigate the protective role of the seed coat under saline conditions. Waxes in black seed coats were more abundant than those in brown seed coats. Salinity (500 mM NaCl) increased the concentration of Na⁺ and decreased the concentration of K⁺ in both black and brown seeds regardless of chloroform treatment. Chloroform treatment alone (in the absence of NaCl) had no effect on the concentration of Na⁺ or K⁺ in black or brown seeds and in the presence of 500 mM NaCl had no effect on the concentration of Na⁺ or K⁺ in brown seeds. However, chloroform treatment increased Na⁺ and decreased K⁺ in black seeds with 500 mM NaCl. A change of MDA (malondialdehyde) concentration in black and brown seeds treated with or without chloroform was similar to the change of Na⁺ concentration. High salinity (1500 mM NaCl) pretreatment for 40 days had a less adverse effect on germination of black seeds compared with brown seeds after they were transferred to fresh water regardless of chloroform treatment. Similar results were found for seedling emergence. In conclusion, a black seed coat may be more protective than a brown seed coat, probably by shielding the embryo from ion toxicity, because of its higher content of waxes. Thus black seeds can better maintain seed viability than brown seeds for extended periods under hypersaline conditions.     

Prediction of desiccation sensitivity in seeds of woody species: a probabilistic model based on two seed traits and 104 species

BACKGROUND AND AIMS: Seed desiccation sensitivity limits the ex situ conservation of up to 47 % of plant species, dependent on habitat. Whilst desirable, empirically determining desiccation tolerance levels in seeds of all species is unrealistic. A probabilistic model for the rapid identification of woody species at high risk of displaying seed desiccation sensitivity is presented. METHODS: The model was developed using binary logistic regression on seed trait data [seed mass, moisture content, seed coat ratio (SCR) and rainfall in the month of seed dispersal] for 104 species from 37 families from a semi-deciduous tropical forest in Panamá. KEY RESULTS: For the Panamanian species, only seed mass and SCR were significantly related to the response to desiccation, with the desiccation-sensitive seeds being large and having a relatively low SCR (i.e. thin 'seed' coats). Application of this model to a further 38 species, of known seed storage behaviour, from two additional continents and differing vegetation types (dryland Africa and temperate Europe) correctly predicted the response to desiccation in all cases, and resolved conflicting published data for two species (Acer pseudoplatanus and Azadirachta indica). CONCLUSIONS: This model may have application as a decision-making tool in the handling of species of unknown seed storage behaviour in species from three disparate habitats.     

Deposition and localization of lipid polyester in developing seeds of Brassica napus and Arabidopsis thaliana

Mature seeds of Arabidopsis thaliana and Brassica napus contain complex mixtures of aliphatic monomers derived from non-extractable lipid polyesters. Most of the monomers are deposited in the seed coat, and their compositions suggest the presence of both cutin and suberin layers. The location of these polyesters within the seed coat, and their contributions to permeability of the seed coat and other functional properties are unknown. Polyester deposition was followed over Brassica seed development and distinct temporal patterns of monomer accumulation were observed. Octadecadiene-1,18-dioate, the major leaf cutin monomer, was transiently deposited. In contrast, the saturated dicarboxylates maintained a constant level during seed desiccation, whereas the fatty alcohols and saturated omega-hydroxy fatty acids continually increased. Dissection and analysis of Brassica seed coats showed that suberization is not specific to the chalaza. Analysis of the Arabidopsis ap2-7 mutant suggested that suberin monomers are preferentially associated with the outer integument. Several Arabidopsis knockout mutant lines for genes involved in polyester biosynthesis (att1, fatB and gpat5) were examined for seed monomer load and composition. The variance in polyester monomers of these mutants is correlated with dye penetration assays. Furthermore, stable transgenic plants expressing promoter::YFP fusions showed ATT1 promoter activity in the inner integument, whereas GPAT5 promoter is active in the outer integument. Together, the Arabidopsis data indicated that there is a suberized layer associated with the outer integument and a cutin-like polyester layer associated with the inner seed coat.     

A suite of sucrose transporters expressed in coats of developing legume seeds includes novel pH-independent facilitators

A suite of newly discovered sucrose transporter genes, PsSUF1, PsSUF4, PvSUT1 and PvSUF1, were isolated from the coats of developing pea (Pisum sativum L.) and bean (Phaseolus vulgaris L.) seeds. Sequence analysis indicated that deduced proteins encoded by PsSUF1, PvSUT1 and PvSUF1 clustered in a separate sub-group under sucrose transporter Clade I, whereas the deduced protein encoded by PsSUF4 clustered in Clade II. When expressed in yeast, these genes were shown to encode sucrose transporters with apparent Michaelis Menten constant (Km) values ranging from 8.9 to 99.8 mm. PvSUT1 exhibited functional characteristics of a sucrose/H+ symporter. In contrast, PsSUF1, PvSUF1 and PsSUF4 supported the pH- and energy independent transport of sucrose that was shown to be bi-directional. These transport properties, together with that of counter transport, indicated that PsSUF1, PvSUF1 and PsSUF4 function as carriers that support the facilitated diffusion of sucrose. Carrier function was unaffected by diethylpyrocarbonate and by maltose competition, suggesting that the sucrose binding sites of these transporters differed from those of known sucrose/H+ symporters. All sucrose transporters were expressed throughout the plant and, of greatest interest, were co-expressed in cells considered responsible for sucrose efflux from seed coats. The possible roles played by the novel facilitators in sucrose efflux from seed coats are discussed.     

Morphology of fruits, seeds and embryos of Argentinian Capparis L. (Capparaceae)

Embryos, seeds and fruits of four species of Capparis L. (Capparaceae) from Argentina are described in this paper: C. flexuosa (L.) L. sensu lato, C. retusa Griseb., C. speciosa Griseb. and C. tweediana Eichl. Chlorophyllous embryos are found in C. flexuosa and C. retusa , species with thin seed coats, dehiscent siliques, and somewhat fleshy red endocarps. Nonchlorophyllous embryos are found in the berry-fruited C. speciosa and C. tweediana , the former with leathery indehiscent fruit wall, the latter with thin-walled dehiscent berries with massive fleshy pulp. The pulp is permeated by seed coat hairs in C. tweediana . The embryos of C. flexuosa , C. retusa and C. tweediana belong to the 'Axile Division, Foliate Subdivision, and Folded Type' of the classification devised by A. C. Martin in 1946. Some features of the embryo of C. speciosa (which are not considered in Martin's classification), support a new subdivision (Massive) and a new morphological type (Unequal cotyledons), characterized by profound anisocotyly, globose shape of the major cotyledon, presence of suberized cells and an undeveloped radicle. Anatomical studies of embryos and seedlings confirm the anisocotyly.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 145 , 209−218.