Seed coat anatomy,karyomorphology, and relationships ofSimmondsia (Simmondsiaceae)

The development of the seed coat ofSimmondsia, whose relationships are extremely problematic, is documented, and its structure is compared to those of putatively related families Euphorbiaceae and Buxaceae. InSimmondsia, the young seed coat is composed of (1) the palisadal exotesta, (2) the thick aerenchymatous mesotesta which is further differentiated into the outer and the inner tissue of mesotesta, and the undifferentiated (3) endotesta and (4) tegmen. At maturity, only the palisadal exotesta composed of thick-walled and prismatic cells, as well as the outer tissue of mesotesta composed of elongate, thick-walled cells, are persistent, while all the remainder is crushed. These distinctive structural features of the exo- and mesotesta inSimmondsia are not found in Euphorbiaceae, but prevalent in Buxaceae. Evidence from seed coat anatomy and other sources supports the view thatSimmondsia has close affinities with Buxaceae, and be placed as a distinct family along with Buxaceae in Buxales.Simmondsia has 2n=52 as a tetraploid ofx=13, and probably represents a taxon adapted to desert areas by polyploidization in the order.     

Seed coat structure and development in Solanum L. section Solanum (Solanaceae)

Seed coat structure in Solanum L. section Solanum (Solanaceae). Seed coat structure in Solanum L. section Solanum was examined using SEM. The seed coat patterns observed were fairly uniform throughout the section, and thus of little diagnostic use. Moreover, all species are characterized by their seeds being covered by hairlike structures, which are bands of lignified thickening in the lateral (and basal) walls of the outer epidermal cells of the testa, and which only become obvious when the seeds are wetted. The development of the seed coat and its characteristic thickening was followed from the unfertilized ovule to the mature seed in vouchered material of Solanum nigrum L. since this species is the generic type, using sections of freshly fixed material.     

The tegumental surface of Phyllodistomum conostomum (Olsson, 1876) (Digenea), revealed by scanning electron microscopy

The external surface of P. conostomum is characterized by relatively large ridges encircling the anterior part of the worm at regular intervals. On the posterior part depressions on the ventral side at regular intervals and relatively small ridges on the dorsal side are present. Ventroposteriorally cobblestone-like protuberances observed are arranged in longitudinal rows. No corresponding arrangement was found dorsally. Only domed papillae (with and without a central knob) were observed, tentatively identified as sensory organs. The regular pattern of these papillae on the ventral and oral sucker is described, in addition to their arrangement ventrally and at the anterior end. A frontal pit anterior of oral suckers and a notch at the posterior end are figured and briefly described. No spines were observed on the body tegument.     

Comparative phytochemical analysis of wild and in vitro-derived greenhouse-grown tubers, in vitro shoots and callus-like basal tissues of Harpagophytum procumbens

Comparative phytochemical analysis of wild and in vitro-derived greenhouse-grown tubers, in vitro shoots and callus-like basal tissues of Harpagophytum procumbens was done. Dried samples were ground to fine powders and their total iridoid (colorimetric method), phenolic [Folin-Ciocalteu (Folin C) method] and gallotannin (Rhodanine assay) contents as well as anti-inflammatory activity [cyclooxygenase assays (COX-1 and COX-2)] were determined. The tissue culture-derived tubers had the highest total iridoid content which was significantly higher than that of the tubers collected from the wild and other tissue cultured materials evaluated. This suggests that cultivated plants can be a viable alternative source of the active principle(s). The total phenolic and gallotannin contents of the wild tubers were significantly higher than the tissue culture-derived tubers and other in vitro-derived plant materials. The presence of phenolic compounds including gallotannins in the tissue cultured materials is of interest from a pharmacological point of view given the pharmacological role of phenolics. In general, extracts from wild tubers demonstrated better inhibitory activities in both COX-1 and COX-2 assays when compared to the tissue culture-derived tubers. All the petroleum ether (PE) and dichloromethane (DCM) extracts showed moderate (50-70%) to good (> 70%) inhibitory activities whereas the ethanol (EtOH) extracts showed poor or no inhibition in both assays. Based on previous reports indicating weak inhibition of COX-2 enzyme by harpagoside, the inhibitory activities of both COX enzymes exhibited by PE and DCM extracts in the current study could be due to the presence of other constituents in the extracts. This points towards the need to identify other active constituents and evaluate their role and mode of action in relation to harpagoside — the main active principle.     

Seed structure and germination in buriti (Mauritia flexuosa), the Swamp palm

Associations of recalcitrance and dormancy are rare, and little information is available concerning the structure of seeds demonstrating this type of behavior or their ecological implications. Mauritia flexuosa is a palm tree associated with swampy environments in the Amazon rainforest and areas of Cerrado (neotropical savanna), the latter biome having marked climatic seasonality. We describe the structures and physiological aspects of the seeds and seedlings of this species to examine its adaptations to swampy environments and its germination control mechanisms, as well as the relationship between recalcitrance and dormancy in terms of reproductive success. Morphoanatomy and histochemistry of the seeds and seedlings were evaluated using standard methodologies, and the effects of temperature and operculum removal on germination were investigated. Differentiated tracheal elements in the embryo are associated with recalcitrance, and presence of numerous stomata on the embryo and the abundance of secondary metabolite compounds in the seedling indicate their adaptations to swampy environments. The inability of the embryo to overcome the resistance of the adjacent tissues determines dormancy of the physiological type, thus the removal of the operculum is an efficient method for promoting propagation. Cellular elongation in the cotyledon promotes seedling protrusion. Mobilization of endosperm reserves is achieved by symplastic flux and is associated with overcoming of dormancy by weakening the tissues adjacent to the embryo. The association between recalcitrance, which favors adaptation to swampy environments, and dormancy, which favors dispersal, is crucial for the reproductive success of this species and its broad geographical distribution.     

The unusual tegumental tissues of the Lunaria annua (Brassicaceae) seed: A developmental study using light and electron microscopy

With light, fluorescence, transmission electron, and environmental scanning electron microscopy we studied the development of the Lunaria annua L. (Brassicaceae) seeds in order to reveal basic anatomical information about the unusual tissues of these seeds. In particular the seed tegument tissues possess complex morphological aspects that are relevant to the biology and ecology of this plant. A sclerenchymatic tissue as the innermost layer of the teguments apparently offers robust protection for the embryo, yet is organized to be flexible. This tissue likely controls the passage of water from the tegumental layers towards the embryo. We report here the presence of tannins in the pre-sclerenchymatic layer of the unripe seed. The inner tegument also houses a spongy tissue with wide intercellular spaces. This tissue could impart buoyancy to the seeds, which possibly might be required for water transport. The structural features could indicate that Lunaria may have evolved in a Mediterranean environment, which is characterized by a long dry season, but with a large amount of rainfall concentrated in short periods. Probably, not only the typical enlarged and flattened fruits of Lunaria can easily float and be dispersed away from the mother plant, but also the seeds have this dispersal peculiarity after release from the silicules.     

Light microscopy and scanning electron microscopy of the In vitro evagination process of Echinococcus multilocularis protoscolices

Marchiondo A. A. and Andersen F. L. 1984. Light microscopy and scanning electron microscopy of the in vitro evagination process of Echinococcus multilocularis protoscolices. International Journal for Parasitotogy14:151–157. During histogenesis of the protoscolices of Echinococcus multilocularis, the apical portion of the protoscolex consisting of the suckers, rostellum and hook region develops as an introversion and invagination within the tissue of the basal portion. In vitro incubation of protoscolices in evagination fluid stimulates the emergence of the apical portion. The initiation of evagination is first detected by a surface change in the basal portion. The smooth contour of this surface which lacks microtriches becomes transformed into tegumental indentations that form transverse and longitudinal furrows within the basal tegument as the protoscolices contract and expand, respectively. An orifice formed at the site or junction where the apical portion is invaginated begins to expand laterally in order to allow emergence of the suckers. The hooks are arranged within the invaginated protoscolex with blades directed towards the basal orifice, the handles directed towards the peduncle and the guards directed laterally. This arrangement persists throughout the evagination of the suckers and rostellum until the apical dome of the hook region emerges, thereby rotating the blades laterally in the direction of the peduncle and rotating the handles and guards medially to assume a coronal arrangement. Evagination is an asynchronous event and therefore allows observation of individual protoscolices in various stages of emergence.     

Fine structure of spermatozoa of Chondrostoma nasus and Rutilus meidingerii (Teleostei, Cyprinidae), as revealed by scanning and transmission electron microscopy

Fürböck, S., Patzner, R.A. and Lahnsteiner, F. 2008. Fine structure of spermatozoa of Chondrostoma nasus and Rutilus meidingerii (Teleostei, Cyprinidae), as revealed by scanning and transmission electron microscopy. — Acta Zoologica (Stockholm) 91 : 88–95
The fine structure of spermatozoa of sneep or nase, Chondrostoma nasus , and lake chub, Rutilus meidingerii , was investigated by means of scanning and transmission electron microscopy. The uniflagellate spermatozoa of C. nasus lacked an acrosome. The flagellum contained the conventional nine peripheral doublets and one central pair of microtubules (9 + 2 pattern) and lacked lateral fins. The uniflagellate spermatozoa of R. meidingerii were made up of a head, also without an acrosome. For both species the sperm tail was covered by a plasma membrane. The midpiece of C. nasus contained five or six mitochondria on average, vesicles and glycogen granules, whereas the midpiece of R. meidingerii had seven mitochondria of a spherical or ovoid shape. The centriolar complex was located caudolaterally with respect to the nucleus. In C. nasus , the centrioles were orientated at an angle of 125° to each other, whereas the centrioles of R. meidingerii were at an angle of 110°. The fine structure of C. nasus and R. meidingerii spermatozoa showed species-specific differences in the position of the proximal centriole relative to the distal centriole, the position and number of mitochondria, size of the head and the length of the flagellum. (Correction added on 11 June 2009, after first online publication: The word 'axoneme' was deleted from the sentence 'The flagellum contained the conventional nine peripheral doublets and one central pair of microtubules (9 + 2 pattern) axoneme and lacked lateral fins.')     

Early developmental stages of two actinosporeans,Raabeia and Aurantiactinomyxon (Myxozoa), as detected by light and electron microscopy

The development of actinosporeans in their oligochaete host proceeding pansporocyst formation is relatively well documented, however, phases preceding it are not as well known. The initial stages in the development of two actinosporeans, Raabeia type 1 of Oumouna et al. [Parasitol. Res. 2002] and Aurantiactinomyxon pavinsis (Ormières, 1968) Marquès [Languedoc, Universite des Sciences et Techniques, Dissertation, 1984] from schizogony to gametogony and sporogony are described. Both actinosporeans begin their development as multinucleate stages near the basal lamina of the oligochaete intestine. Proximal to these stages and between the host epithelium cells are uninucleate cells whose nuclei divide to produce binucleate cells. These divide mitotically to produce cells with four nuclei which then undergo plasmotomy to yield a tetracellular stage and the first phase in pansporocyst formation. From the uninucleate stage to the tetranucleate stage, the cell membrane of the parasite is associated closely via finger-like projections with the intestinal epithelial and glandular cells of the host.     

河静黑叶猴果实性食物组成、选择及其对种子的扩散作用

喀斯特森林的生态环境较其他类型的森林更为脆弱.由于抗干扰和恢复能力低,喀斯特森林中种子扩散对维持植物种群的续存与更新具极其重要的作用.叶猴以叶为主要食物,果实是仅列其后的第二大食物资源.对分布于喀斯特生境的叶猴来说,它们对果实的取食具有潜在的种子传播作用.基于此,于2009年7月-2010年12月,在越南广平的风芽-格邦国家自然公园,通过实地跟踪观察结合粪便收集调查了河静黑叶猴(Trachypithecus francoisi hatinhensis)对果实的选择及其对种子的潜在扩散作用.河静黑叶猴全年共采食果实类食物38科131种,取食的果实种类和取食高峰持续期均高于其他地区黑叶猴对果实的取食.这得益于当地原始雨林中丰富的果实食物资源.河静黑叶猴对果实种类没有明显的选择偏好(-0.3＜S112种果实＜0.3),果实资源的可获得性亦不影响其对果实的取食频次(r=-0.13,P=0.15＞0.05),在雨季的果实食物种类和取食强度上均高于旱季(z=-2.903,P=0.02＜0.05).在果实性状的选择上,河静黑叶猴对处理难度较小的浆果和核果(两种类型的果实共106种,占80.9％)有明显的选择偏好,更多地选择黄色(46种)、红色(29种)和绿色(14种)(三种颜色的果实占果实种类的68.7％),重量多为5 g以上较大的果实(共104种,占果实种类的79.4％),更多地选择果实内仅有1-2颗种子的果实.河静黑叶猴多吞食直径小于3 cm的种子,在猴群的粪便中发现了85种果实的种子,尤以肉质的浆果和核果最多,如垂叶榕(Ficus benjamina)、柿叶青梅(Vatica diospyroides)、构树(Broussonetia papyrifera)等.最远传播距离可达397 m.对其他的果实,则以吐出或丢弃的方式短距离传播,多集中在6-20 m范围.河静黑叶猴是一些果实较大或果皮较硬的植物重要的潜在扩散媒介,因为此类种子不能依靠鸟类吞食传播,如水浦桃(Syzygium jambos)、红毛丹(Nephelium lappaceum)和木奶果(Baccaurea ramiflora)等.河静黑叶猴一年中在雨季的夏秋两季(8-12月)对果实的取食强度达到高峰,这期间每月在猴群粪便中发现的种子数量均超过1000.结果反映出河静黑叶猴对喀斯特森林中的果实植物种子有潜在的传播作用.     

Fracture of plant tissues and walls as visualized by environmental scanning electron microscopy

The environmental scanning electron microscope (ESEM) provides a highly relevant and controllable environment in which to study hydrated systems without the artefacts of other highly prepared specimens. The instrument facilitates control of turgor through hydration using different chamber vapour pressures. Deformation of a simple plant tissue-upper epidermal layers in Allium cepa (onion)-was observed at the scale of the two principal failure mechanisms: cell breakage; and cell separation induced by treatment with a chelating agent. Cell rupture and release of contents occurred at cellular junctions ahead of an imposed growing notch, indicating that disruption of cells occurred remotely from the creation of a new surface. Cells that separated usually maintained their turgor and the separation process took place through progressive failure of middle lamellar material seen as strands between separating cells. These mechanisms were compared with the rupture of excised Chara corallina walls that occurred by formation and breakage of strands between separating wall layers. This study provides in situ visual characterization of wall rupture and cell separation at the microscopic level in hydrated plant material.     

Comparison of surface topography of three species of Diphyllobothrium (Cestoda,pseudophyllidea) by scanning electron microscopy

Plerocercoids of different sizes as well as adult worms of D. dendriticum, D. latum and D. ditremum were studied using scanning electron microscopy (S.E.M.). In the plerocercoids there were found distinct differences in appearance and length of microtriches between these three species, while the microtriches of adult worms were more similar. A regional difference in microtrix appearance was found in the larvae of D. ditremum and D. dendriticum. This was not apparent with S.E.M. in adult worms. The length of ‘body’ microtriches in D. dendriticum varied with the length of the larvae. The topography of the genital atrium of mature and gravid proglottids in adult worms of these three species is also described.     

Cell wall extension results in the coordinate separation of parallel microfibrils: evidence from scanning electron microscopy and atomic force microscopy

Enlargement of the cell wall requires separation of cellulose microfibrils, mediated by proteins such as expansin; according to the multi-net growth hypothesis, enlargement passively reorients microfibrils. However, at the molecular scale, little is known about the specific movement of microfibrils. To find out, we examined directly changes in microfibril orientation when walls were extended slowly in vitro under constant load (creep). Frozen-thawed cucumber hypocotyl segments were strained by 20-30% by incubation in pH 4.5 buffer or by incubation of heat-inactivated segments in alpha-expansin or a fungal endoglucanase (Cel12A). Subsequently, the innermost layer of the cell wall was imaged, with neither extraction nor homogenization, by field-emission scanning electron microscopy (FESEM) and atomic force microscopy (AFM). AFM images revealed that sample preparation for FESEM did not appreciably alter cell wall ultrastructure. In both FESEM and AFM, images from extended and non-extended samples appeared indistinguishable. To quantify orientational order, we used a novel algorithm to characterize the fast Fourier transform of the image as a function of spatial frequency. For both FESEM and AFM images, the transforms of non-extended samples were indistinguishable from those of samples extended by alpha-expansin or Cel12A, as were AFM images of samples extended by acidic buffer. We conclude that cell walls in vitro can extend slowly by a creep mechanism without passive reorientation of innermost microfibrils, implying that wall loosening agents act selectively on the cross-linking polymers between parallel microfibrils, rather than more generally on the wall matrix.     

A study by scanning (SEM) and transmission (TEM) electron microscopy of the glomerular capillaries in developing rat kidney

Summary Kidneys of 2 to 10 day-old rats of Wistar and Sprague-Dawley strains were fixed with glutaraldehyde by retrograde vascular perfusion and then prepared for observation in TEM and SEM. In addition methacrylate casts of differentiating glomerular capillaries were examined by SEM. Although the glomerular vascular pattern differs from one glomerulus to another, its differentiation proceeds according to the following general plan. First the glomerular capillary splits longitudinally, finally to form 3 to 5 lobules consisting of a capillary network, sustained centrally by the mesangium.In the present study the differentiation of glomerular capillaries is described in five successive arbitrarily selected stages. At Stage I a capillary loop penetrates between the lower limb and the middle segment of the S-shaped body, the rudimentary nephron. At Stage II the capillary undergoes a first subdivision, establishing the primitive lobulation of the glomerulus. At Stage III the vascular and urinary poles differentiate. At Stage IV the glomerulus assumes the aspect of a spherical body, and the capillaries in each lobule undergo subdivision. In Stage V the glomerular vascular pattern approaches its adult appearance, although the maturation processes continue for an extended period of time. Hence in the 10 day-old rat the best-differentiated glomeruli are half the size of adult glomeruli, and their capillary loops are proportionally less well-developed. The capillaries of adjacent lobules may communicate with each other, but a direct vascular shunt between the afferent and efferent vessels cannot be demonstrated.Presented in part at the 60ème Congrés de l'Association des Anatomistes in Nice (27–30 May 1976) and the 73. Versammlung der Anatomischen Gesellschaft in Innsbruck (12–15 September 1978)The author thanks Miss Morena Ghisletta for excellent technical assistance     

Morphological studies on the periostracum of the fresh-water mussel Amblema (uniondae): Light microscopy, transmission electron microscopy, and scanning electron microscopy

The structure of the periostracum in the fresh-water mussel Amblema has been described using light microscopy, transmission elec;ron microscopy, and scanning electron microscopy. The structure and evolutive course of the periostracum was studied along its entire length, from the periostracal groove until it forms the tough outer covering of the shell. At least five structurally and functionally distinct regions were identified. In addition, the periostracum itself was seen to be a multilayered structure consisting of three major layers which are themselves subdivided into minor layers. From these morphological observations, a regulatory role for the various periostracal layers in mineral trapping, nucleation, and the subsequent formation of the prismatic and nacreous layers of the shell can be postulated.     

Mode of infection of the corn earworm (Heliothis zea) by Beauveria bassiana as revealed by scanning electron microscopy

Conidia from highly pathogenic mutants of Beauveria bassiana germinate quickly (within 18 hr) on the surface of corn earworm larvae (Heliothis zea) and immediately begin penetration of the cuticle. Enzymes produced by the penetrating hyphae degrade the cuticle since holes are formed at the point of entry. Clustering of conidia around nodules of larvae is often seen, but penetration is not restricted to such areas. Direct evidence is presented to show that conidia can also germinate inside of spiracle openings and could invade larvae by this route. Once inside the hemocoel, the fungus multiplies extensively; however, larval death occurs with only minimal breakdown of internal tissues. During mummification, outgrowths of fungal hyphae occur first and most extensively in the intersegmental regions of larvae. Conidia from mutants exhibiting low levels of pathogenicity are either significantly delayed or do not germinate on larval surfaces. When germination does occur, hyphae from such mutants do not penetrate immediately; instead, extensive surface hyphal growth, with or without eventual penetration of the integument, is evident.     

Embryo structure,seed traits,and productivity of relict vine Aristolochia contorta (Aristolochiaceae)

The herbaceous vine Aristolochia contorta (Aristolochiaceae) is a rare plant with a fragmented area in East Asia. Confocal laser-scanning microscopy and stereomicroscopy were employed to examine the seed and embryo structure. This is the first research on embryo anatomy in Aristolochia. Shape and structure of this embryo may be phylogenetically significant. The seed structure of A. contorta is similar to that of other Aristolochia species. Considerable variation is found for productivity traits; the average seed set is rather low (26.7%). The studied seed and embryo traits of A. contorta are found to be variable, but may be potentially informative at the genus and species level. The restricted productivity and reduced population size threaten the continued survival of A. contorta and, perhaps, the butterfly that depends on it.     

Fasciolagigantica: Parasitological and scanning electron microscopy study of the in vitro effects of ivermectin and/or artemether

Objective

To evaluate the in vitro effects of different concentrations of ivermectin and/or artemether on Fasciolagigantica worms and to study the parasitological changes and tegumental alterations using scanning electron microscopy (SEM).

Methods

Fasciola gigantica worms were incubated in vitro for 24 and 48 h with three concentrations of either ivermectin or artemether (10, 20 and 50 μg/ml) or both in half concentration of either (5, 10 and 25 μg/ml).

Results

Exposure of Fasciola worms to 25 + 25 μg/ml of combined drug regimens or to 50 μg/ml of either ivermectin or artemether for 48 h led to 100%, 41.7% and 75% worm killing which were accompanied by a significant reduction in egg laying capacity and significant increase in dead eggs maximally recorded in combined drug regimens. SEM of the flukes incubated for 48 h with combined drug regimens showed maximal tegumental disruption with swelling of the worm body, roughness, blebbing, sloughing and complete loss of spines. Disruption to the tegument of the flukes induced by artemether was more than that of ivermectin.

Conclusions

Artemether alone or combined with ivermectin in half doses had potent fasciocidal activities. Besides, half doses of combined drug regimens had higher ovicidal effects than each drug alone. In vivo studies are recommended to explore the efficacy of combined regimens against Fasciola infection.     

Seed coat carotenoids of the cycad genera Dioon,Encephelartos, Macrozamia and Zamia: Evolutionary significance

Seed coat carotenoids of Dioon, Encephelartos, Macrozamia and Zamia with ranges in Africa, the West Indies, Australia and Mexico are simple mixtures which conform to the theoretical primitive angiosperm “magnolian pattern”. Lycopene is the principal pigment of Zamia while Encephelartos, Dioon and Macrozamia coats contain a mixture of the unsubstituted, mono-and di-hydroxy-β-carotenes. The evolutionary significance of these results is discussed.