Cold-acclimation of Hibiscus rosa-sinensis L. and Hibiscus syriacus L. in natural and controlled environments

Abstract Seasonal cold-acclimation patterns and the effects of photoperiod and temperature on cold-hardiness of Hibiscus rosa-sinensis L. and Hibiscus syriacus L. were determined. Field-grown H. rosasinensis consistently failed to survive freezing at - 2°C. Two genotypes of field- and container-grown H. syriacus initiated cold-acclimation in mid September, in response to decreasing daylength, and continued to an ultimate midwinter hardiness level of - 27°C in early February. Controlled environment experiments using combinations of short days (SD) and cool day/night temperatures were unable to induce even minimal cold acclimation of H. rosasinensis. In controlled environments, H. syriacus attained a moderate amount of cold tolerance at warm temperatures and long days (LD). Low night temperature combined with LD, warm day produced the same degree of cold-acclimation as the SD treatments. While not essential, SD enhanced H. syriacus cold-acclimation in controlled environments. A - 5°C frost treatment of intact plants did not enhance cold-hardiness of H. syriacus.     

Occurrence of cotton aphids (Aphis gossypii) and lady beetles (Hamonia axyridis) on Hibiscus syriacus Linne: Are the aphids a pest of cucurbits?

Populations of cotton aphid on Hibiscus syriacus increased rapidly from 17 to 24 May 2007, and then decreased as its predator, the lady beetle Hamonia axyridis, increased in number. There was a 10 day time lag between peak populations of aphids and lady beetles. The infestation of aphids on H. syriacus produced some damage, but H. syriacus recovered soon after the lady beetles arrived. Cotton aphid clones from H. syriacus were transferred to other summer host plants: to five different vegetables on two dates, and to cucumber on three dates. Apart from one case where reproduction occurred on eggplant, most H. syriacus aphid clones did not survive on the vegetables. The cotton aphid on H. syriancus was prey and a food source for H. axyridis and acted to conserve natural enemies.     

Effects of various chemical agents and early ethylene production on floral senescence of Hibiscus syriacus L.

To understand the factors that induce floral senescence in Hibiscus syriacus L., we have investigated the effects of various chemical agents on flower senescence at two different flowering stages, before and after full bloom, as well as the relationship between flower longevity and endogenous ethylene production before full bloom. Treatments with ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC), and ethephon enhanced floral senescence, while aminoethoxyvinylglycine (AVG) promoted flower longevity regardless of treatment timing. Although ethanol slightly extended flower longevity, abscisic acid (ABA), nitric oxide, boric acid and sucrose, which have been reported to affect flower longevity or senescence, had no effect on H. syriacus floral senescence. The polyamine spermine (SPM), methylglyoxal-bis(guanylhydrazone) (MGBG), an inhibitor of SPM biosynthesis, and cycloheximide (CHI) accelerated flower senescence when applied before full bloom, but had no effect when applied after full bloom. SPM, MGBG and CHI treatments resulted in enhanced ethylene production during flower opening, and the promotion of flower senescence is mediated by ethylene production prior to full bloom. Furthermore, endogenous ethylene, spontaneously produced before blooming, was closely associated with floral senescence. These results suggest that ethylene production during flower opening plays a key role in determining the timing of Hibiscus flower senescence.     

Gas chromatography-mass spectrometric analysis of oxidative degradation products of sporopollenin inMagnolia grandiflora (Magnoliaceae) andHibiscus syriacus (Malvaceae)

Sporopollenin can be dissolved in water by oxidative degradation with using potassium permanganate (KMnO₄). in mature pollen grain ofMagnolia grandiflora andHibiscus syriacus some organic compounds were extracted from the aqueous phase into hexane, and analyzed by using gas chromatography-mass spectrometry (GC-MS). The analysis shows that the extracted compounds Include some organosilicon compounds. By researching of mass spectra data base, the organosillicon compounds are suggested as 1, 1, 1, 5, 7, 7, 7-heptamethyl-3,3-bis(trimethylsilox))tetrasiloxane and 1, 1, 1, 3, 5, 7, 7, 7-octamethyl-3, 3- bis(trimethylsiloxy)tetrasiloxane. 1,2-benzendicarboxylic acid butyl 2-ethylhexyl ester is also suggested by data base research in the extracted compounds. The present study implies that silicon (Si) is considered to be located in inner part of three dimensional structure of sporopollenin.     

Isolation and culture of protoplasts from callus tissue of Hibiscus syriacus L.

Protoplasts were isolated from callus tissue of Hibiscus syriacus L. using a solution of 3% Onozuka cellulase, 1% Onozuka macerozyme, and 0.5% hemicellulase. Highest yields of viable protoplasts were obtained from friable, white or yellow callus 8–9 days after subculture on Murashige & Skoog medium with 0.5 mg l^-1 2,4-dichlorophenoxyacetic acid and 0.1 mg l^-1 kinetin. Protoplasts cultured in thin liquid layers of this medium with mannitol continued dividing for longer than those cultured in droplets or in an agar medium. Cultures were maintained until protoplasts had divided to form groups of more than ten cells. Cell groups developed into callus and continued to grow on an agar medium, but failed to differentiate on a regeneration medium with 2 mg l^-1 naphthalene acetic acid and 1 mg l^-1 benzylaminopurine.     

Chloroplast genome of Hibiscus rosa-sinensis (Malvaceae): Comparative analyses and identification of mutational hotspots

Previous studies to resolve phylogenetic and taxonomic discrepancies of Hibiscus remained inconclusive. Here, we report chloroplast genome sequence of Hibiscus rosa-sinensis. Hibiscus rosa-sinensis chloroplast genome was 160,951 bp, comprising of large single copy (89,509 bp) and small single copy (20,246 bp) regions, separated by IRa and IRb (25,598 bp each). The genome contained 130 genes including 85 protein-coding genes, 37 transfer RNAs and 8 ribosomal RNAs. Comparative analyses of chloroplast genomes revealed similar structure among 12 species within family Malvaceae. Evolutionary rates of 77 protein-coding genes showed 95% similarities. Analyses of codon usage, amino acid frequency, putative RNA editing sites, and repeats showed a great extent of similarities between Hibiscus rosa-sinensis and Hibiscus syriacus. We identified 30 mutational hotpots including psbZ-trnG, trnK-rps16, trnD-trnY, trnW-trnP, rpl33-rps18, petG-trnW, trnS-trnG, trnH-psbA, atpB-rbcL, and rpl32-trnL that might be used as polymorphic and robust markers to resolve phylogenetic discrepancies in genus Hibiscus.     

Prevailing insects in Roselle plants,Hibiscus sabdariffa L., and their efficiency on pollination

Field surveys of Roselle plant fields were conducted during two successive seasons of 2006 and 2007. The total counted insects on Roselle, Hibiscus sabdariffa attained 16 species belong to seven insect Orders [(Hemiptera (3 species), Homoptera (1 species), Lepidoptera (3 species), Neuroptera (1 species), Coleoptera (3 species), Diptera (1 species), Hymenoptera (4 species)]. Variable numbers of insects were counted on different Roselle varieties and in the two seasons. The most prevailing insect species were the piercing sucking pests [Empoasca spp.; Spilostethus longulus Dallas; Nezara viridula L. and Oxycarenus hyalinipennis (Costa)] and the predators [Polistes sp., Coccinella undecimpunctata and Scymnus syriacus]. Honeybee (Apis mellifera L.) proved to be the most abundant pollinator, averaging 13, 17, 18 individuals/25double strikes in cultivations of Sudani, Masri and White varieties of Roselle plants during flowering of the first season respectively. Similar results were also obtained in the second season. The highest number of honeybees was recorded around 10 am to midday during the flowering stage for the two seasons. It was obvious that the effects of the two hydrothermal, factors temperature and relative humidity, had slight or negligible effects on the occurrence of insect pollination throughout the two seasons of the study, while statistical analysis of the obtained results showed that the productivity of Roselle plant (mean number of seeds and mean weights of seeds) were significantly affected by pollinators.     

Growth and behaviour of six species of amphibian larvae in a winter pond in Israel

The effect of different ecological niches on growth and behaviour of larvae of four frogs (Bufo viridis, Hyla arborea, Pelobates syriacus, Rana ridibunda), and two salamanders (Salamandra salamandra and Triturus vittatus) found in a winter pond was studied. S. salamandra, T. vittatus, R. ridibunda and B. viridis were found most of the time on the bottom of the pond. However, H. arborea tadpoles were found throughout the pond and were usually sedentary, as compared with P. syriacus which moved up and down constantly. S. salamandra, T. vittatus, R. ridibunda and R viridis tadpoles from the bottom of the pond grew faster than the tadpoles from the surface of the pond. However, tadpoles of H. arborea and P. syriacus growing at the bottom or on the surface developed at similar rates.The invertebrate biomass increases during the summer and was higher at the bottom of the pond than at the surface. However the amount of chlorophyll a was about the same at the surface and at the bottom of the pond. S. salamandra and T. vittatus tadpoles feed on various types of of invertebrates, R. ridibunda and H. arborea and B. viridis tadpoles feed on vascular plants and algae, and P. syriacus tadpoles feed on both invertebrates and plants.     

Cytogenetic characteristics and the breeding system in six Hibiscus species

Cytogenetic characteristics confirm that Hibiscus acetosella and Hibiscus cannabinus are outbreeders, while Hibiscus asper, Hibiscus physaloides, Hibiscus sabdariffa and Hibiscus surattensis have evolved into inbreeders. The inbreeding species appear to have co-evolved a floral structure in which some anthers abut on the stigma prior to anthesis. Received: 19 February 1999 / Accepted: 17 June 1999     

Specialized host-plant performance of the cotton aphid is altered by experience

Although distinct host specialization is observed for the cotton-melon aphid (Aphis gossypii Glover) on cotton and cucurbit plants, it is still ambiguous whether the specialization is altered by experience on a novel host plant. Here the performance of cotton and cucurbit-specialized aphids, A. gossypii on novel host plants was studied by a host-selection test and by the life-table method. The two host-specialized aphids cannot survive and establish populations after reciprocal host transfers. They have ability to recognize the host plants on which they were reared, and escape behavior from novel hosts was observed. Interestingly, the cotton and cucurbit-specialized aphids survive and reproduce normally on hibiscus (Hibiscus syriacus), a main overwintering host plant, and host-fidelity of A. gossypii to cucurbit plants is altered by feeding and living experience on hibiscus, which confers the same capacity to use cotton and cucumber on to the cucurbit-specialized population, but host-fidelity to cotton is not altered and the fitness of the cotton specialized population to cucumber is still poorer. A. gossypii from hibiscus has a significant preference for cotton to cucumber in the host-selection process, and none stays on cucumber more than 20 h after transfer. The results presented imply that cucurbit-specialized aphids might not return to an overwintering host plant (hibiscus) in wild fields, so host conservatism to cucurbit plants is maintained. The potential of cucurbit-specialized aphids of A. gossypii to use cotton plants, intermediated by experience on hibiscus, suggests that the specialized host-plant performance of phytophagous insects is not wholly conservative.     

ONTOGENETIC DEVELOPMENT OF SPINOUS EXINE IN HIBISCUS SYRIACUS (MALVACEAE)

Pollen development in Hibiscus syriacus L. (Malvaceae) was studied with light (LM), scanning (SEM) and transmission (TEM) electron microscopes, with special attention to the formation of extremely long spines of the pollen grains. At the early tetrad stage, probacules are initiated directly on the plasma membrane and grow in coincidence with the height of primexine matrix within a callosic wall. Subsequently, a pretectum appears at the top of the probacules and then a foot layer is formed by accumulation of white line centered lamellations. Before dissolution of the callosic wall, a reticulate patterned pretectum is established around the microspores. There is not, however, any morphological indication on the initiation of the spines during the tetrad period within a callosic wall. It is after dissolution of the callosic wall that the spines of exine begin to form by the apposition of lamellated sheets. The lamellated sheets show a concentric configuration around the developing supratectal spines. The mature pollen grain is spheroidal, polycolporate, 160–170 μm in diameter, with supratectal spines 20–25 μm long. The supratectal spines of Hibiscus pollen are not homologous with the other exinous protrusions which are determined within the callosic wall during tetrad stage.     

Chemical Composition and Antimicrobial Activity of Satureja,Thymus, and Thymbra Species Grown in Lebanon

The essential oils were extracted by hydrodistillation from Satureja cuneifolia, Satureja thymbra, Coridothymus capitatus, Thymus syriacus, and Thymbra spicata growing wild in Lebanon. Their phytochemical analysis performed by GC/MS showed that the aforementioned species are characterized either by carvacrol (60.9%) or thymol (54.3%) or by a more or less equal amounts of these two phenols. Assessment of their in vitro antimicrobial activity against Candida albicans and six pathogenic bacteria using the broth dilution method revealed that the tested oils have a broad activity spectrum with minimum inhibitory concentration ranging from 0.1 to 0.8 mg/ml. Among the tested species, S. thymbra EO showed the highest antimicrobial potential whereas T. syriacus showed the lowest inhibitory activity. These results give scientific evidence for the use of those species in the Lebanese folk medicine and lend support to implement them as natural alternatives for synthetic antimicrobials.     

Anthocyanins isolated from Hibiscus syriacus L. attenuate lipopolysaccharide-induced inflammation and endotoxic shock by inhibiting the TLR4/MD2-mediated NF-κB signaling pathway

BackgroundHibiscus syriacus L. has been used as a medicinal plant in many Asian countries. However, anti-inflammatory activity of H. syriacus L. remains unknown.PurposeThis study was aimed to investigating the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. variety Pulsae (PS) on the lipopolysaccharide (LPS)-induced inflammation and endotoxic shock.Study design and methodsMTT assay and flow cytometry analysis were performed to determine cytotoxicity of PS. RT-PCR, western blotting, and ELISA were conducted to evaluate the expression of proinflammatory mediators and cytokines. Molecular docking study predicted the binding scores and sites of PS to TLR4/MD2 complex. Immunohistochemical assay was conducted to evaluate the binding capability of PS to TLR4/MD2 and nuclear translocation of NF-κB p65. A zebrafish endotoxic shock model was used to evaluate anti-inflammatory activity of PS in vivo.ResultsPS suppressed LPS-induced nitric oxide and prostaglandin E₂ secretion concomitant with the downregulation of inducible nitric oxide synthase and cyclooxygenase-2 expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as TNF-α, IL-6, and IL-12 in LPS-stimulated RAW 264.7 macrophages. Additionally, molecular docking data showed that PS mostly fit into the hydrophobic pocket of MD2 and bound to TLR4. In particular, apigenin-7-O-glucoside powerfully bound to MD2 and TLR4 via hydrogen bonding. Additionally, immunohistochemistry assay revealed that PS inhibited LPS-induced TLR4 dimerization or expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation, resulting in the inhibition of NF-κB activity. PS also attenuated LPS-mediated mortality and abnormality in zebrafish larvae and diminished the recruitment of neutrophils and macrophages at the inflammatory site accompanied by the low levels of proinflammatory mediators and cytokines.ConclusionPS might be a novel immunomodulator for the effective treatment of LPS-mediated inflammatory diseases.     

Phenological responses of plants to climate change in an urban environment

Global climate change is likely to alter the phenological patterns of plants due to the controlling effects of climate on plant ontogeny, especially in an urbanized environment. We studied relationships between various phenophases (i.e., seasonal biological events) and interannual variations of air temperature in three woody plant species (Prunus davidiana, Hibiscus syriacus, and Cercis chinensis) in the Beijing Metropolis, China, based on phenological data for the period 1962–2004 and meteorological data for the period 1951–2004. Analysis of phenology and climate data indicated significant changes in spring and autumn phenophases and temperatures. Changes in phenophases were observed for all the three species, consistent with patterns of rising air temperatures in the Beijing Metropolis. The changing phenology in the three plant species was reflected mainly as advances of the spring phenophases and delays in the autumn phenophases, but with strong variations among species and phenophases in response to different temperature indices. Most phenophases (both spring and autumn phenophases) had significant relationships with temperatures of the preceding months. There existed large inter- and intra-specific variations, however, in the responses of phenology to climate change. It is clear that the urban heat island effect from 1978 onwards is a dominant cause of the observed phenological changes. Differences in phenological responses to climate change may cause uncertain ecological consequences, with implications for ecosystem stability and function in urban environments.     

Influence of chemical profiles of host plants on the infestation diversity ofRetithrips syriacus

The onset of biotic stress in the host plants as a result of increased insect population size leads to enhanced levels of secondary metabolites and associated phenolic enzyme activity. Of the three host plants examined, viz.Ricinus communis (castor),Eucalyptus globulus (eucalyptus) andManihot utilissima (tapioca), castor was the host most preferred byRetithrips syriacus. Despite the fact that tapioca had the highest levels of secondary compounds, thrips infestation persisted. However, fecundity and growth were reduced because of the relatively high levels of primary metabolites. Gallic acid was found to be the most toxic of the phenolic acids, followed by pyrogallol, resorcinol, phloroglucinol and vanillic acid. The less toxic phenolic acids and flavanoids were detected in leaves that harboured thrips, while the preponderance of gallic acid was found in uninfested hosts. Thus the interaction ofRetithrips syriacus with the hosts is governed essentially by the biochemical profiles of its hosts, which tend to be altered subsequent to infestation, thus manifesting induced resistance through enhanced production of phenolics     

Ecological Plasticity of the Photosynthetic Apparatus of <Emphasis Type="Italic">Hibiscus syriacus</Emphasis> L. under Pressure of High Temperature,Insolation, and Air Pollution

This paper describes the integrated effect of high summer temperature, intensive insolation, gas pollution, and dust in the air on the pigment content and net oxygen production (apparent photosynthesis) and dark respiration rates of common hibiscus (Hibiscus syriacus L.) leaves. The study included three observation sites: Tashkent Botanical Garden (Academy of Sciences, the Republic of Uzbekistan), the public garden in the central part of the city of Tashkent, and a mountain holiday camp. The research was carried out in 2017 during the period of active vegetation. The results of experiments showed a high adaptive potential of H. syriacus L., as well as its adaptiveness to stress environmental factors of the semiarid zone, provided a sufficient level of its irrigation. The ecological plasticity of the photosynthetic apparatus of hibiscus plays a key role in species adaptation to environmental conditions. It has been revealed that the growth of the plants of the Tashkent Botanical Garden under shading conditions leads to the formation of large, wide, thin, shade leaf blades, which can be considered as the manifestation of sciomorphosis. Heliomorphosis features of hibiscus leaves were identified in mountains, where sun leaves with significantly smaller, thickened, and compacted blades are formed under high insolation. The adaptive significance of these structural leaf modifications is to strengthen photosynthetic capacity for compensating the deficiency in sunlight (in the case of sciomorphosis), and, on the contrary, to provide mutual shading for photosynthetic elements as a measure of protection against the damaging effect of redundant solar radiation (in the case of heliomorphosis). This provides carbon dioxide assimilation and organic matter production for maintaining the constant energy balance for the plant under different environmental conditions. The study of the temperature correlation of dark respiration and net oxygen production rates has shown that these processes are more resistant to temperature injuries under more extreme environmental conditions. In addition, mature H. syriacus leaves have a higher resistance than young leaves; i.e., the plant adapts to possible temperature drops in the process of its ontogenesis. Taking into account the optimal measuring conditions, the net oxygen production rate of H. syriacus during the period of active vegetation is approximately at the same level under different growing conditions (0.20 ± 0.05 μmol of O₂/(dm² s)); this is considered a norm of reaction of the net production (apparent photosynthesis) rate of H. syriacus and determines the specific features of its photosynthetic apparatus.     

Comparative life table statistics for the ladybeetle Scymnus syriacus reared on the green citrus aphid,Aphis spiraecola,fed on two host plants

Demographic statistics for the ladybeetle Scymnus syriacus Marseul (Coleoptera: Coccinellidae) were investigated when reared on the green citrus aphid, Aphis spiraecola Patch (Homoptera: Aphididae), fed on two host plants: spirea (Spirea sp.) and Thompson orange (Citrus sinensis cv. Thompson) under laboratory conditions (25 ± 1°C, 65 ± 5% relative humidity and 16 h light : 8 h dark). Mean developmental times from egg to adult were 20.32 ± 0.18 and 22.07 ± 0.15 days for spirea and Thompson orange, respectively. The survival rate from egg to adult was higher for spirea (85%) than for Thompson orange (72.5%). The oviposition periods were 41.8 ± 2.47 and 39.71 ± 2.6 days and the total number of eggs per female were 657.6 ± 29.5 and 587.9 ± 32.6 on spirea and Thompson orange, respectively. Fertility life tables were also constructed to compare the effects of host plants. The intrinsic rate of increase (r_m) on spirea was higher than that on Thompson orange. Jackknife estimates of r_m varied from 0.149 on spirea to 0.133 on Thompson orange. The mean population generation times on these host plants were 38.16 and 40.65 days, respectively. These results indicate that spirea‐fed A. spiraecola is more suitable for S. syriacus, producing a higher survival rate and shorter developmental time.     

Details of cavities used by Syrian Woodpeckers Dendrocopos syriacus in Hungary

ABSTRACT

Eighty-two nest and roosting cavities of Syrian Woodpeckers Dendrocopos syriacus were documented over a period of 25 years (1994–2018). Fourteen different tree species were used, the majority being non-native in Hungary. Cavities were found in both main trunks and in larger limbs. The median cavity height was 3.0 m and ranged from 1 to 7 m. The cavity entrance orientation was non-random with a preference for a south-southeast orientation.     

Bemerkungen zum Maurenspecht,Picoides major numidus

Zusammenfassung Voous (1947) meinte, der BlutspechtPicoides syriacus und eine gut gekennzeichnete nordafrikanische Unterart des BuntspechtsP. m. numidus seien die Nachfahren von Rückzugs-populationen des Buntspechtes während der Mindel-Eiszeit. Alle morphologischen Merkmale, die sonst zur Unterscheidung von Blut- und Buntspecht herangezogen werden, zeigen, daßP. m. numidus ein Buntspecht ist. Dasselbe trifft für die Lautäußerungen zu. Im Gegensatz zu europäischen Rassen weistP. m. numidus einen ökologischen Sexualdimorphismus auf. gehen öfter in der Kronenschicht der Nahrungssuche nach. Sie halten sich dabei auch öfter an dünnen Zweigen auf als . Kein deutlicher Unterschied besteht bezüglich der Tätigkeiten des Nahrungserwerbs. Ökologisch ist die nordafrikanische Rasse mitteleuropäischen Buntspechten ähnlicher als Blutspechten. In der Erörterung des ökologischen Sexualdimorphismus wird auf das inselartige Vorkommen, die Eintönigkeit des Lebensraumes, das offensichtliche Fehlen einer Nahrungsspezialisierung im Winter, Merkmalsfreisetzung und die Rolle der Geschlechter im Fortpflanzungsgeschehen eingegangen. Die Auffassung wird vertreten, daßP. m. numidus keine unmittelbaren historischen Beziehungen zuP. syriacus hat.

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Remarks onPicoides major numidus

Summary Voous (1947) proposed that the Syrian WoodpeckerPicoides syriacus and an distinct North African subspecies of the Great Spotted WoodpeckerPicoides major numidus are the descendants of refugial populations of the Great Spotted Woodpecker during the Mindel glaciation. All morphological characters commonly used for discerningP. major andP. syriacus show thatP. m. numidus belongs to the former species. The same is true for the vocalizations. In contrast to European subspeciesP. m. numidus exhibits an ecological sexual dimorphism. Females feed more often in the crown of a tree. They also forage more often on thin twigs than males do. No significant difference between the feeding techniques of male and female was found. The North African subspecies is more similar ecologically to Central European Great Spotted Woodpeckers than to Syrian Woodpeckers. In discussing the ecological sexual dimorphism the insular distribution, the low habitat diversity, the apparent lack of a specialized winter food, competitive release and the role of the sexes in the breeding cycle are considered. It is pointed out that this situation does not allow to explain the dimorphism by the sole action of one of these factors. It is suggested thatP. m. numidus has no direct historical relationship withP. syriacus.

     

A SEASONAL STUDY OF THE VEGETATIVE SHOOT APEX AND THE PATTERN OF PITH DEVELOPMENT IN HIBISCUS SYRIACUS

Tolbert , Robert J. (West Virginia U., Morgantown.) A seasonal study of the vegetative shoot apex and the pattern of pith development in Hibiscus syriacus. Amer. Jour. Bot. 48(3): 249–255. Illus. 1961.—The shoot apex of Hibiscus syriacus L. is described as having a cytohistological zonation superimposed on a tunica-corpus configuration. The apex is flat-topped or may have a saddle-back or concave appearance as seen in median longitudinal section. The metrameristem, consisting of the tunica and corpus initials, is comprised of large, light-staining, vacuolate cells that have thick cell walls and exhibit much dark-staining intercellular substance. Surrounding the metrameristem is the flanking meristem, which is responsible for the outer layers of the shoot, and from which the leaf primordia arise. The pith rib meristem lies below the metrameristem and consists of files of cells that are responsible for the pith. There are no major seasonal changes in the structure of the apex during the yearly cycle. The pith displays a long-shoot type of development with the cells remaining in distinct files during the first flush of growth in the spring. As growth slows and internode elongation is gradually reduced, the pith displays the characteristic short-shoot type of development, consisting of a spongy tissue of rounded cells with many intercellular spaces and no distinct files of cells. A crown is differentiated across the top of the pith at the end of the growth period. This consists of a band of cells with thick, dark-staining cell walls, which separates by the apex from the last year's growth. In contrast to many gymnosperms, this crown is dispersed by renewed cell activity the following spring.