Exine development: the importance of looking through a colloid chemistry ``window'

Most biological construction systems operate within the colloidal dimension. In view of this, it seems reasonable to reassess what is known of the early stages of exine development in the light of a brief excursion into colloid and micelle behaviour. The results of this analysis show remarkable similarity of structures and suggest that almost all of the features seen during early pollen wall development can be easily interpreted using simple, established colloidal principles. This study of exine framework and endexine development offers the possibility that growth of the early exine progresses by successive transitory mesophases of a constrained micellar system. The self-assembling micelle mesophases will all be clearly recognized as constituents of the developing exine. They include spherical, cylindrical, continuous layers of hexagonally-packed cylindrical units and lamellar mesophases which most probably correspond to future granules, columellae, complex columellar (and alveolar) microarchitecture and ``white-line-centred' lamellae. Furthermore, the various types of micelle involved have the potential to perform the functions previously loosely assigned to the exine.     

Pollen wall development in Hypecoum imberbe Sm. (Fumariaceae)

We have studied the ontogeny of the pollen wall of Hypecoum imberbe. Our initial observations with conventional transmission electron microscopy (TEM) were inconclusive due to the similarities found in the electrodensity of the foot layer and the endexine, and between this latter layer and the intine. Thus we applied the PTA acetone histochemical test in order to differentiate between the cell wall components. This method proved to be efficient in resolving the differences between the layers and even allowed us to distinguish two strata within the narrow intine layer. A thin foot layer can be distinguished only by the temporary presence of a single white line separating it from the initial deposition of the endexine. The aperture consists of two colpi with no special differentiation. The tapetum is a typical secretory type giving rise to elaioplasts and tapetosomes during development, which persist as individual organelles upon the degradation of the tapetum. As the ultrastructural organisation of the sporoderm seems to offer little protection to the gametophyte, we finally discuss how the structure of the pollen sporoderm might be related to the special morphological characteristics of a flower, its habitat, and the biology of the plant as a whole.     

Pollen morphology in the genus Veronica L. (Plantaginaceae) and its systematic significance

Pollen grains of 30, mainly annual, species from 134 populations of the genus Veronica (Plantaginaceae, formerly Scrophulariaceae) from the Mediterranean Region, have been studied with light and scanning electron microscopy. Three pollen types are defined based on pollen size and exine surface sculpture. In some cases within the study group pollen size can be useful in species determination. Hypothetical relationships of pollen grain size and aperture number with ploidy level, style length and corolla diameter are discussed. The ornamentation of the exine in Veronica, although generally a conservative character within the genus, gives some additional support to the most recent infrageneric classification of Veronica.     

Pollen, tapetum and orbicule development in Modiolastrum malvifolium (Malvaceae)

Background and Aims: Although orbicular functions are still a matter of debate, theyare considered by most authors to be exclusively formed by asecretory tapetum. However, the presence of orbicules on a peritapetalmembrane associated with a plasmodial tapetum has been describedfor Abutilon pictum (Malvaceae) in a previous study. Thus, studieson other species of Malvaceae are necessary to corroborate thepresence of such bodies in other members of the family. Pollenand microsporangium development of Modiolastrum malvifoliumhas been studied in this work. Methods: Anthers at different stages of development were processed fortransmission electron microscopy and light microscopy. Membranesand pollen walls resistant to acetolysis were isolated fromwhole anthers. Key Results: Microspore tetrads have a tetrahedral arrangement. Pollen grainsare shed at the bicellular stage. The tapetum is invasive, non-syncytialand a peritapetal membrane with orbicules is formed. Conclusions: This is the first report of the presence of orbicules on a peritapetalmembrane in a species with a tapetum of an invasive, non-syncytialtype. Taking into consideration all the information on the subject,it can be concluded that the presence of orbicules is not astable criterion to differentiate between a secretory or plasmodial,or intermediate invasive, non-syncytial tapetum.     

Establishment of Plantago lanceolata L. and Plantago major L. among grass

Summary Establishment of Plantago lanceolata and P. major ssp major among grass was studied in a field experiment in which survival and selection on date of seedling emergence and plant size was investigated in relation to the vegetation structure. P. major — in contrast to P. lanceolata — was not able to establish itself in grass because of its lower competitive ability caused by later germination, smaller seedling size, and shorter leaves. In both species there was selection for early germination. For P. lanceolata a significant correlation was found between the strength of selection and the light climate, determined by the structure of the grass sward. Plants that germinated early were at an advantage because they were larger, especially the leaves, when compared with plants that germinated late. It seems likely that selection was mainly by competition for light. Contrary to expectation P. major-seedlings had a higher shade tolerance than those of P. lanceolata. The performance of both species is discussed in relation to their different life strategies.Grassland species research group publication no 142     

Plasmodial tapetum and pollen wall development in Butomus umbellatus (Butomaceae)

This report describes the ultrastructural development of plasmodial tapetum and pollen wall of Butomus umbellatus. The tapetum contains extensive arrays of rough endoplasmic reticulum, vesicles from which are responsible for the formation of sporopollenin-like bodies. The tapetum is also involved in the formation of other forms of sporopollenin precursors. Development of pollen wall continues after microspores are released from their callosic walls; they are then enclosed by plasmodial tapetum. The activity and products of the plasmodial tapetum show substantial correlation with pollen wall development, particularly ektexine formation. In B. umbellatus, the tapetum intrudes into the anther locule at early microspore stage. This timing of plasmodial intrusion occurs at a later stage of pollen development as compared to those in the advanced monocotyledons. We report the rough endoplasmic reticulum origin of sporopollenin-like bodies and their occurrence in the plasmodial tapeta of B. umbellatus.     

Influence of restoration on arbuscular mycorrhiza of Biscutella laevigata L. (Brassicaceae) and Plantago lanceolata L. (Plantaginaceae) from calamine spoil mounds

The arbuscular mycorrhizal status of two plant species, Biscutella laevigata L. and Plantago lanceolata L., was investigated on calamine spoil mounds in Boles?aw (southern Poland). Although B. laevigata is a member of the Brassicaceae, a family generally accepted as non-mycorrhizal, this species formed AM symbioses on both heavy metal-contaminated and non-contaminated sites. Besides vesicles and coils, arbuscules were also observed, especially in roots collected prior to seed maturity. Relative mycorrhizal root length and relative arbuscular richness were usually much higher in P. lanceolata than in B. laevigata but not absolute arbuscule richness. Roots of P. lanceolata showed higher colonisation than B. laevigata. Although roots were collected from plants in close proximity, no correlation in mycorrhizal parameters was found between the two species.     

Influence of restoration on arbuscular mycorrhiza of Biscutella laevigata L. (Brassicaceae) and Plantago lanceolata L. (Plantaginaceae) from calamine spoil mounds

The arbuscular mycorrhizal status of two plant species, Biscutella laevigata L. and Plantago lanceolata L., was investigated on calamine spoil mounds in Boles^aw (southern Poland). Although B. laevigata is a member of the Brassicaceae, a family generally accepted as non-mycorrhizal, this species formed AM symbioses on both heavy metal-contaminated and non-contaminated sites. Besides vesicles and coils, arbuscules were also observed, especially in roots collected prior to seed maturity. Relative mycorrhizal root length and relative arbuscular richness were usually much higher in P. lanceolata than in B. laevigata but not absolute arbuscule richness. Roots of P. lanceolata showed higher colonisation than B. laevigata. Although roots were collected from plants in close proximity, no correlation in mycorrhizal parameters was found between the two species.     

Pollen wall ultrastructure and ontogeny in Heliotropium europaeum L. (Boraginaceae)

The pollen grains of Heliotropium europaeum are heterocolpate, with alternation of 3 colpori and 3 pseudocolpi. The exine is characterized by a scabrate and thick tectum, massive columellae with a granular appearance and a thick nexine. The thickening of the intine at the apertural level makes the interpretation of this zone difficult. The ontogenetic study helped to understand the ultrastructure of the exine and the apertures. The different steps are as follows. The primexine matrix is formed during the beginning of the tetrad stage; it consists of an outer thick and electron dense zone and an inner one, less dense to electrons. The tectum and the infratectum begin to form in the outer zone of the matrix, towards the middle of the tetrad stage. The infratectum consists of a network of columellae variable in thickness and oriented in different directions. The foot layer is lacking. The endexine is formed on a lamella system during the callose loss and microspore separation. The endexine becomes compact very early on its inner part. The apertures are initiated during the tetrad stage; a granulo-fibrillar oncus develops. At the free microspore stage, the oncus gets fibrillar and is bordered by endexine lamellae on its outer side and by endexine granulations on its inner one and laterally. The intine is set at the end of this stage. At the vacuolated microspore stage, the intine shows three layers: two thin, clear and homogeneous layers, one outside and the other inside, and a thick middle layer that forms the zwischenkörper, crossed by trabecula, in the apertural areas.     

Pollen wall development in Cryptomeria japonica (Taxodiaceae)

Pollen wall development in Cryptomeria japonica was observed by scanning and transmission electron microscopy. The pollen of C. japonica is characterized by a non-saccate, projecting papilla. The exine of C. japonica consists of the outer granular ectexine and the inner lamellated endexine. At the tetrad stage, the initial granular layer of the pro-ectexine first forms on the microspore plasma membrane. The tripartite lamellae of the pro-endexine form under the pro-ectexine. The prosporopollenin material is deposited on the pro-ectexine and pro-endexine at the free spore stage. The ectexine granule increases its volume and the endexine lamellae thicken. The papilla protrudes during the tetrad stage. The tip of the papilla bends laterally where the exine is thinner. Exine construction in C. japonica is similar to that of Cunninghamia; however, the amount and size of the granular ectexine and lamellated endexine differ. The conspicuous papilla protrudes and bends during the tetrad period.     

Carbon integration in Plantago aristata (Plantaginaceae): the reproductive effects of defoliation

Patterns of carbon integration in aclonal species are poorly understood in spite of their potential to influence individual fitness. To provide more information about these patterns, we performed a defoliation experiment with P. aristata. We examined, at the metameric level, the reproductive responses to the removal of the major carbon sources within metamers. Bracts on marked reproductive spikes and leaves subtending these spikes were removed at three stages of reproductive maturity: spike elongation, flowering, and fruiting. Spike dry weight and length, capsule number, seeds per capsule, and seed weight were measured. We tested the hypothesis that seed weight would respond least to defoliation. We also performed a complementary ¹⁴C translocation experiment to measure the amount of radioactive carbon moving into the marked spikes from outside the metamer. Defoliation depressed all components of reproduction within marked spikes, and little ¹⁴C was translocated from outside the metamer into the reproductive spikes, even those that were defoliated. Both results support the view that reproductive metamers in this species are largely autonomous with respect to their carbon budget. Defoliation during spike elongation most depressed reproduction, and bract removal depressed reproduction more than did leaf removal. The data suggest that bracts compensate for leaf removal by increasing their photosynthetic rate; however, the ability to compensate differs among plant populations. Of all the reproductive components, seed weight was least affected by defoliation. The data show, however, that the time of defoliation relative to reproductive development influences which reproductive components are affected.     

Pollen wall development in Malvastrum corchorifolium (Malvaceae)

The Malvaceae family has many species that are the subject of systematic controversy. Its broad spectrum of fruit and pollen morphology has challenged taxonomists because often morphological studies lead to different conclusions. Therefore, this study of selected stages of pollen wall development in Malvastrum corchorifolium is offered as another approach to verify its current taxonomic assignment and assist in the clarification of malvacean ancestry. Aperture number, endexine thickness and the presence of basal cushions are features that concur with its present placement in the Abutileae tribe, Sidinae subtribe and genus Malvastrum. Its advanced spine morphology, long spines with pointed apices and prominent basal cushions, suggest that Malvastrum corchorifolium is highly specialized and evolutionarily advanced.     

Pollen wall development in Sciadopitys verticillata (Sciadopityaceae)

Pollen wall development of Sciadopitys verticillata was observed by transmission electron microscopy. The pollen of S. verticillata is non-saccate and spherical, and the exine consists of the outer thick, sculptured ectexine and the inner lamellated endexine. At the early tetrad stage, the initial ectexine and lamellae of the initial endexine begin to form on the microspore plasma membrane. The ectexine granules gradually swell. Deposition of sporopollenin materials on the ectexine granules then results it their becoming partially connected to each other. Identification of the original small ectexine granules then becomes difficult, and, finally, the ectexine appears as a homogeneous, partially discontinuous layer. The granules of the early ectexine cannot be identified. At maturity, there are four to five endexine lamellae. Recent molecular data have shown that Sciadopitys first branches off from the Cupressaceae plus Taxaceae clade, which is characterized by granular exine. Although the ectexine of Sciadopitys is similar to that of the Cupressaceae during initial development, the morphology of the ectexine is significantly different in the mature pollen. The initial stage of pollen development clearly shows the structural homology of the granular ectexine. Divergence of the exine structure occurs in the later stages.     

Early ontogenetic patterns in chemical defense in Plantago (Plantaginaceae): genetic variation and trade-offs

Predictions based on the plant age and growth-differentiation balance hypotheses of defense were tested in two congeneric species, Plantago lanceolata and P. major, by quantifying iridoid glycosides, defensive chemicals, in seeds and leaves during the first 6 wk of growth. Concentrations decreased from the seed to 2-wk-old seedling stage in P. lanceolata, but increased during this period in P. major. In both species, levels were similar for 2- and 4-wk-old plants, then significantly increased from 4 to 6 wk. Genetic variation in the ontogeny of iridoid glycoside production was significant in both species at the maternal family level and at the population level. To examine whether allocation costs could explain the low production of iridoid glycosides in seedlings, relationships between growth and defense (iridoid glycosides) were characterized. Growth and defense had a positive or null relationship in all age groups, indicating that there was no trade-off in these plants at any age. This study provides some support for the growth-differentiation balance hypothesis, but offers no support for the plant age hypothesis. Measuring how herbivory affects plant fitness at different ontogenetic stages may shed light on these patterns in Plantago and on the evolution of the ontogeny of defense.     

Pollen competition reduces inbreeding depression in Collinsia heterophylla (Plantaginaceae)

We tested two predictions of the hypothesis that competition between self-pollen may mitigate negative genetic effects of inbreeding in plants: (1) intense competition among self-pollen increases offspring fitness; and (2) pollen competition reduces the measured strength of inbreeding depression. We used Collinsia heterophylla (Plantaginaceae), an annual with a mixed mating system, to perform controlled crosses in which we varied both the size of the pollen load and the source of pollen (self vs. outcross). Fitness of selfed offspring was higher in the high pollen-load treatment. Our second prediction was also upheld: inbreeding depression was, on average, lower when large pollen loads were applied (11%) relative to the low pollen-load treatment (28%). The reduction was significant for two fitness components relatively late in the life-cycle: number of surviving seedlings and pollen-tube growth rate in vitro. These findings suggest that intermittent inbreeding, which leads to self-fertilization in plants with genetic loads, may select for traits that enhance pollen competition.     

Reproductive allocation and resource compensation in male-sterile and hermaphroditic plants of Plantago lanceolata (Plantaginaceae)

Gynodioecy is a breeding system in which hermaphrodites coexist with male steriles. Theoretical models predict that without any compensation in female fitness male steriles will disappear from a population due to their reproductive disadvantage. In the present study I investigated whether male-sterile (MS), partially male-sterile (IN), and hermaphroditic (H) plants of Plantago lanceolata differed in reproductive growth and allocation. Offspring of three interpopulation crosses segregating all three sex morphs were grown under nitrogen-limited conditions in a growth chamber. Independent of the genetic background MS plants attained a higher vegetative and reproductive dry mass and a higher reproductive output than H plants, whereas IN plants had intermediate values. When corrected for the mass of the pollen, the dry mass differences between the sex morphs were much reduced but still present. However, when whole-plant allocation was expressed on the basis of nitrogen, the differences between the sex morphs disappeared. Thus the sex morphs took up similar amounts of nitrogen but distributed them differently. The MS and IN plants used the nitrogen saved by not producing pollen for additional vegetative as well as reproductive growth. The data presented in this study suggest that resource compensation is one of the main mechanisms responsible for the maintenance of MS and IN plants in gynodioecious P. lanceolata.     

Pollen wall development in Eucommia ulmoides (Eucommiaceae)

In Eucommia the foot layer plays a prominent part in microspore wall development. Bacules (>100 nm) are rods originating within the foot layer. Small bacules (diameter ca. 10 nm) form at the same time as the foot layer. The tectum is considered to be made up of these microbacular rods. Spinules ar continuous with the bacules. An endexine is differentiated during a middle to late microspore stage. Except for the pore the furrow includes tectum and foot layer on the endexine. Since the furrow consists of a region of reduced foot layer and the reduction is gradual near the polar ends of furrows, assessment of furrow length depends to some extent upon variations in exine infolding. The pore is well defined, but, because it is crossed by lamellations of the endexine and foot layer and often overlaid by tongues and bridges of foot layer plus tectum (including spinules) it is obscured from view using either light microscopy or scanning electron microscopy.     

Patterns of leaf asymmetry changes in Plantago major L. (ssp. major) natural populations exposed to different environmental conditions

In the present study, developmental stability of leaf traits was examined in three natural populations of Plantago major L. (ssp. major), representing two polluted environments (Karaburma and Zemun) and an unpolluted area (Crni Lug). Developmental stability was assessed as fluctuating asymmetry (FA). The magnitude of FA is believed to reflect differences in the ability of individuals to buffer their development in natural populations. We hypothesized that there are differences within characters and among characters in response to environmental conditions. Significant patterns of asymmetry correlations and asymmetry changes were detected both within characters and between characters. The manova results revealed a significant effect of individual and a significant individual × environment interaction on actual asymmetry (logL_i ? logR_i) and on the amount of asymmetry |(logL_i ? logR_i)| for leaf width and vein distances within a leaf. Over time, statistically significant and positive correlations of the FA values were detected for each trait separately per sample (population). For both leaf traits, there were differences for (logL_i ? logR_i) and |(logL_i ? logR_i)| asymmetry values among individuals within samples in response to yearly variations. Statistically significant and negative correlations for (logL_i ? logR_i) versus |(logL_i ? logR_i)| asymmetries were detected for both leaf traits. In summary, our results highlight the importance of differences in the ability of individuals to buffer their development under different environmental conditions and point to the concept that developmental stability is character specific.