Crinoid holdfasts from the Silurian of Gotland

Crinoid holdfasts occur throughout the Silurian sequence of Gotland, with a marked concentration to reefs and immediate reef surroundings. Four morphological groups are recognized: (1) Discoid holdfasts. (2) Cirriferous holdfasts, represented by (a) Rhizocrinus -like holdfasts, (b) large radices with stout, often branching cirri, and (c) rhizoid holdfasts, formed by a complicated net of pseudocirri. (3) Creeping stems attached to the substrate by thin strands of stereome. (4) Coiling stems. Discoid attachments, cirriferous holdfasts (types b and c) and coiling stems show little preference to substrate and adapt morphologically to conditions at hand. Rhizoid holdfasts display the greatest variation and apparently occur within many different groups of crinoids. Rhizocrinusttke holdfasts have been found only in quiet-water deposits, while creeping stems were concentrated to more turbulent environments. Coiling stems were epizoic, attached to rugose corals, bryozoans, crinoid stems and similar supports. Growth, mode of life (attached or free), settling and fixation of larvae, and relation to substrate and other organisms ate discussed.     

Characterization of the Adhesive Holdfast of Marine and Freshwater Caulobacters

Caulobacters are prosthecate (stalked) bacteria that elaborate an attachment organelle called a holdfast at the tip of the cellular stalk. We examined the binding of lectins to the holdfasts of 16 marine Caulobacter strains and 10 freshwater species or strains by using a panel of fluorescein-conjugated lectins and fluorescence microscopy. The holdfasts of all the marine isolates bound to only wheat germ agglutinin (WGA) and other lectins that bind N-acetylglucosamine (GlcNac) residues. The freshwater caulobacters showed more variability in holdfast composition. Some bound only to WGA and comparable lectins as the marine strains did. Others bound additional or other lectins, and some did not bind to the lectins tested. The binding of WGA appeared to involve the regions of the holdfast involved with adhesion; a holdfast bound to WGA was significantly less adhesive to glass. Competition experiments with WGA-binding holdfasts and oligomers of GlcNac demonstrated that trimers of GlcNac (the preferred substrate for WGA binding) were more effective than dimers or monomers in preventing WGA binding to holdfasts, suggesting that stretches of contiguous GlcNac residues occur in the WGA-binding holdfasts. In addition, differences between freshwater and marine holdfasts in the strength of WGA binding were noted. The effect of a number of proteolytic and glycolytic enzymes on holdfast integrity was examined; the proteases had no effect for all caulobacters. None of the glycolytic enzymes had an effect on marine caulobacter holdfasts, but chitinase and lysozyme (both attack oligomers of GlcNac) disrupted the holdfasts of those freshwater caulobacters that bound WGA. Despite some similarity to chitin, holdfasts did not bind Calcofluor and no measurable effects on holdfast production were detectable after cell growth in the presence of diflubenzuron or polyoxin D, inhibitors of chitin synthesis in other systems. Finally, the holdfasts of all caulobacters bound to colloidal gold particles, without regard to the coating used to stabilize the gold particles. This binding was stronger or more specific than WGA binding; treatment with colloidal gold particles prevented WGA binding, but the reverse was not the case.     

Sunlight-induced bark formation on current-year stems of Euphorbia plants from South Africa

Species of Euphorbia from South Africa have sunlight-induced bark injuries on stems visually identical to sunlight-induced injuries on more than twenty species of long-lived columnar, candelabra cactus species of the Americas. Injury ratios on stems on both groups of plants were similar to ratios of direct sunlight exposure on stem surfaces averaged throughout the annual cycle at many latitudes. Injuries on Euphorbia species occurred on current-year stem tissues. Stem tissues of Euphorbia species have a single-celled epidermis. Some species had a 2- to 3-celled hypodermis. Sunlight-induced bark formation only involved anticlinal followed by periclinal cell divisions of epidermal cells. No other cells were involved in bark formation. Many Euphorbia species have crests and troughs while others have prominent tubercles. Experimental results showed that prominent crests provided shading to concave troughs on some Euphorbia species and these species had less sunlight-induced barking. In contrast, other Euphorbia species that do not have prominent crests have little self-shading and these species had more sunlight-induced barking. Sunlight-induced bark formation occurred during a one year of sunlight exposure on Euphorbia stems while similar barking injuries occur on cactus stems only after several decades of sunlight exposure. These sunlight-induced injuries on young stem tissues of Euphorbia stems may result since current-year stems of Euphorbia plants had thinner cuticles and thinner epidermal/hypodermal layers compared with stem surfaces of cactus plants.     

Growth of Typha latifolia in boreal forest habitats,as measured by double sampling

Seasonal growth of Typha latifolia L. stands in five oxbow lakes of the Athabasca River in the boreal forest zone of Alberta, Canada, was monitored by both the harvest technique and double sampling by regression. The technique for calculating mean biomass and variance for the double sampling procedure is described. The relationship between stem weight and height was linear for flowering stems, but a logarithmic transformation of weight was used for the non-flowering stem regressions. Growth began in late May and stems reached maximum weight by mid-August. Stem mortality (up to 14%) was highest at sites with high stem density, but mortality was limited to small stems at all sites. Total biomass of these stems was less than 4% of the peak above-ground standing crop. In nutrient-poor sites, Typha was found on floating organic mats, but in more nutrient rich sites on grounded substrates. Peak above-ground biomass ranged from 456 to 848 g m^?2, generally in response to the successional and nutrient status of the oxbow lake. Sites returned to an earlier stage of succession by flooding had larger stems and greater biomass.     

Constructing tree stem form from digitized surface measurements by a programming approach within discrete mathematics

Key message

The main message of this work is the demonstration of possibility of creation of stem shape from digitized points using integer-programming approach. The points are digitized by magnetic motion tracker which in contrast to the laser scanning allows the reconstruction of complete cross-section of stem even in the “hidden (invisible)” part.

Abstract

Three-dimensional information on tree stem form plays an important role in understanding the structure and strength of a standing tree against the forces of wind, snow, and other natural pressure. It also contributes to precision in volume measurement compared to conventional two-dimensional measurement. We investigate approaches for obtaining three-dimensional information of tree stem form from partially organized surface measurements, acquired using a three-dimensional digitizing device (Polhemus FASTRAK^® motion tracking device). We then propose a new programming approach from discrete mathematics to construct tree stem form. Our method is based on an optimal connection of neighbor triangles for surface construction, which is created by locally possible combination of three digitized points on the stem surface. We compare the proposed method to the existing heuristic methods of contour tracing and region growing. Our analysis shows that the proposed method provides a consistent construction of tree stem form, for even stems with extremely irregular structure such as those from bent trees and mangrove trees with unique root spread, while the other methods are incapable for constructing such tree stems.     

Radial perforations in crinoid stems from the Silurian of Gotland

Radiating intracolumnal canals are a characteristic feature of large (diameter 10 mm or more) crinoid stems from the Silurian of Gotland. They are found in nodals as well as in internodals where the columnal height exceeds one millimetre. They were formed secondarily in the median and distal portions of crinoid stems with pseudocirriferous holdfasts. Intercolumnal canals are found in the distal parts of stems with true cirri regardless of the size of the stem. It is suggested that these canals played an important role in crinoid physiology. The crinoids are believed to have sustained a large proportion of their tissues through cutaneous digestion and uptake of dissolved substances from the surrounding sea water. The intra- and intercolumnal canals increased the surface of the axial canal in relation to volume. They provided a connection between the axial canal and the surrounding sea water, thus facilitating nutrient transport to the tissues.     

Differences in response between haploid and diploid isomorphic phases of Gracilaria verrucosa (Rhodophyta: Gigartinales) exposed to artificial environmental conditions

This study tests the responses of juvenile gametophytes and tetrasporophytes (holdfast stage) of the isomorphic alga Gracilaria verrucosa under different environmental conditions.Estimations of survival and growth of holdfasts of haploid and diploid juvenile individuals were performed in natural sea-water and artificial culture medium, and under stringent conditions using lead as a toxin and ultra violet radiation as a mutagen. Results indicate that (i) holdfasts of haploid juveniles grow better than diploids in non-optimal medium conditions; (ii) holdfasts of diploid juveniles have a better tolerance to lead than haploids; and (iii) slight advantage of holdfasts of diploid juveniles grow better than haploids under U.V. radiation.     

Patients with impaired verb-tense processing: do they know that yesterday is past?

This paper begins with a focus on the task of stem inflection, where participants are given a verb stem and asked to produce the verb''s past-tense form, which can produce a neuropsychological double dissociation with respect to regular versus irregular verbs. Two differing theoretical interpretations are outlined: one is based on specifically morphological and separate brain mechanisms for processing regular versus irregular verbs; the other argues that the two sides of the dissociation can arise from one procedure, which is not specifically morphological, and which relies to differing extents on phonological versus semantic information for regular versus irregular verbs. We then present data from a different version of the task, in which patients were given past-tense forms and asked to produce the present-tense or stem forms (talked → talk and ate → eat). This change yielded a very different pattern of performance in four non-fluent aphasic patients as a function of the regular–irregular manipulation, an outcome which is argued to be more compatible with the single- than the dual-mechanism account. Finally, we present a small amount of data from a task in which the patient was asked to judge whether spoken regular and irregular verb stems and past-tense forms indicated actions occurring today or yesterday. This task produced an even more different and intriguing pattern of performance suggesting a deficit in morpho-syntactic knowledge: not how to produce past-tense forms but what such forms mean and how that understanding interacts with verb regularity. The paper concludes with a discussion of how the research field of acquired disorders of tense processing might advance as a result of new approaches, in particular those informed by studies of developmental disorders.     

Heterogeneity and Glycan Masking of Cell Wall Microstructures in the Stems of Miscanthus x giganteus,and Its Parents M. sinensis and M. sacchariflorus

Plant cell walls, being repositories of fixed carbon, are important sources of biomass and renewable energy. Miscanthus species are fast growing grasses with a high biomass yield and they have been identified as potential bioenergy crops. Miscanthus x giganteus is the sterile hybrid between M. sinensis and M. sacchariflorus, with a faster and taller growth than its parents. In this study, the occurrence of cell wall polysaccharides in stems of Miscanthus species has been determined using fluorescence imaging with sets of cell wall directed monoclonal antibodies. Heteroxylan and mixed linkage-glucan (MLG) epitopes are abundant in stem cell walls of Miscanthus species, but their distributions are different in relation to the interfascicular parenchyma and these epitopes also display different developmental dynamics. Detection of pectic homogalacturonan (HG) epitopes was often restricted to intercellular spaces of parenchyma regions and, notably, the high methyl ester LM20 HG epitope was specifically abundant in the pith parenchyma cell walls of M. x giganteus. Some cell wall probes cannot access their target glycan epitopes because of masking by other polysaccharides. In the case of Miscanthus stems, masking of xyloglucan by heteroxylan and masking of pectic galactan by heteroxylan and MLG was detected in certain cell wall regions. Knowledge of tissue level heterogeneity of polysaccharide distributions and molecular architectures in Miscanthus cell wall structures will be important for both understanding growth mechanisms and also for the development of potential strategies for the efficient deconstruction of Miscanthus biomass.     

Grow slowly,persist, dominate—Explaining beech dominance in a primeval forest

Being able to persist in deep shade is an important characteristic of juvenile trees, often leading to a strong dominance of shade‐tolerant species in forests with low canopy turnover and a low disturbance rate. While leaf, growth, and storage traits are known to be key components of shade tolerance, their interplay during regeneration development and their influence on juveniles'' survival time remains unclear. We assessed the ontogenetic effects of these three traits on the survival time of beech (Fagus sylvatica), and Norway and sycamore maples (Acer pseudoplatanus, Acer platanoides) in a primeval beech forest. Biomass allocation, age, and content of nonstructural carbohydrates (NSC) were measured in the stems and roots of 289 seedlings and saplings in high‐ and low‐vitality classes. Saplings experienced a trade‐off between absolute growth rate (AGR) and storage (NSC) as the leaf area ratio (LAR) decreases with biomass development. High LAR but low AGR and low NSC corresponded to beech with a marked ability to persist in deep shade while awaiting canopy release. In turn, a comparably small LAR in combination with a high AGR and higher storage (NSC), as observed in Norway maple and sycamore maple, reduced sapling survival time, thus offering an explanation for beech dominance and maple disappearance in the undergrowth of old‐growth beech forests.     

Nouvelles espèces d’Ancyrocrinus et d’Ammonicrinus,crinoïdes à pédoncule spécialisé du Dévonien armoricain (Brest,France)

New species of Ancyrocrinus and Ammonicrinus, crinoids, crinoids with specialized stem from the Armorican Devonian (Brest, France). The discovery of the oldest tetrameral grapnels of Ancyrocrinus, A. crozonensis n. sp., in the Lower Eifelian modifies considerably the previous evolutionary scheme and sets the problem of the transition from tetrameral to pentameral symmetry in the ancyrocrinid holdfasts. The presence of a new species of Ammonicrinus, A. kerdreoletensis n. sp., in the Upper Emsian suggests a North-Gondwanan origin of the genus. Its highly specialized structure of the stem, characteristic of the genus, was developed earlier than previously recognized. To cite this article: J. Le Menn, P.A. Jaouen, C. R. Palevol 2 (2003).     

Functional morphologies of the columns of Upper Ordovician Xenocrinus and Dendrocrinus

Two common Upper Ordovician crinoids, Xenocrinus and Dendrocrinus , had distinctive columns that showed marked contrasts of both form and function. Xenocrinus baeri (Meek) had a tetragonal facet geometry but functioned in the normal manner for a symplectially articulated column. Column flexure was by bending, the four directions of curvature being limited by the tetragonal arrangement of crenulae. In contrast, columns of Dendrocrinus casei Meek, with a characteristic pentastellate symmetry, were able to twist, a functional adaptation not previously reported from crinoids; this was controlled by the unusual geometry of the crenularium. This suggests that some or all crinoid columns may be subject to twisting stresses, perhaps associated with changes in the current direction, that crenulae may resist passively. Functional morphology, column, crinoids, X enocrinus , D endrocrinus     

Restudy of the worm-like carbonaceous compression fossils Protoarenicola,Pararenicola, and Sinosabellidites from early Neoproterozoic successions in North China

The carbonaceous compression fossils Protoarenicola baiguashanensis Wang, 1982, Pararenicola huaiyuanensis Wang, 1982, and Sinosabellidites huainanensis Zheng, 1980, from the early Neoproterozoic Liulaobei and Jiuliqiao formations in northern Anhui, North China, were previously interpreted as worm-like metazoans, largely on the basis of transverse annulations and purported proboscis structures. If correct, these would be some of the earliest known bilaterian animals and would provide a key paleontological calibration to molecular clock analyses. In this study, we examine a large population of these carbonaceous fossils, clarify their taxonomy, and provide new insights into their morphological, paleoecological, and phylogenetic interpretations.Although all three species are characterized by annulated tubes, P. baiguashanensis bears a bulbous terminal structure at one end of its tube. P. huaiyuanensis is characterized by a constricted opening at one end and a closed termination at the other. The two ends of S. huainanensis tubes are both closed and round. The bulbous terminal structure in P. baiguashanensis was previously interpreted as an animal proboscis, but new observations suggest that it was more likely a holdfast structure analogous to discoidal holdfast structures of the Mesoproterozoic Tawuia-like fossil Radhakrishnania Kumar, 2001, and the frondose Ediacara fossil Charniodiscus Ford, 1958. Furthermore, it is possible that at least P. baiguashanensis and P. huaiyuanensis may represent reproductive or taphonomic fragments of the same organism. This reinterpretation weakens the previous interpretation that P. baiguashanensis and P. huaiyuanensis were worm-like bilaterian animals. Instead, they can be alternatively interpreted as erect epibenthic organisms, possibly coenocytic algae reaching a tiering height of 30 mm. The predominance of discoidal holdfasts, as opposed to rhizoidal holdfasts, in pre-Ediacaran epibenthic organisms was probably related to more stable substrates in the presence of microbial mats and in the absence of bioturbating animals.     

Development, distribution, and characteristics of intrinsic, nonbacterial ice nuclei in prunus wood

Ice nuclei active at approximately −2°C and intrinsic to woody tissues of Prunus spp. were shown to have properties distinct from bacterial ice nuclei. Soaking 5-centimeter peach stem sections in water for 4 hours lowered the mean ice nucleation temperature to below −4°C, nearly 2°C lower than stems inoculated with ice nucleation-active Pseudomonas syringae strain B301D. Ice nucleation activity in peach was fully restored by air-drying woody stem sections for a few hours. The ice nuclei in woody tissue were inactivated between 40 and 50°C, but unaffected by treatment with bacterial ice nucleation inhibitors (i.e. NaOCl, tartaric acid, Triton XQS-20), sulfhydryl reagents (i.e. p-hydroxymercuribenzoate and iodine) and Pronase. Ice nuclei could not be dislodged from stems by sonication and were shown to be equally distributed in peach bud and internodal stem tissue on a per unit mass basis; outer and inner stem tissues were also indistinguishable in ice nucleation activity. Development of ice nuclei in immature peach and sweet cherry stems did not occur until midsummer and their formation was essentially complete by late August. Once formed the ice nuclei intrinsic to woody stems were stable and unaffected by seasonal changes in growth. The apparent physiological function of the ice nuclei is discussed in relation to supercooling and mechanisms of cold hardiness in Prunus spp.     

Three-dimensional distribution of mass movement signals in disturbed Picea abies (L.) Karst.

Dendrogeomorphologic approaches enable annual absolute dating of various potentially hazardous geomorphic processes. This study tested the dating precision of three commonly used dendrogeomorphic approaches used for the extraction of geomorphic signals from the tree ring series of disturbed trees. The first approach is based on the onset of reaction wood occurrence, and the other two approaches are based on the detection of abrupt changes in tree ring eccentricity. Nine 165 cm long stems of maximally 27 year old Norway spruces (Picea abies (L.) Karst.) growing on unstable slope affected by mass movement events of known age were analysed. Several sampling directions on stems were analysed to reveal the best sampling strategy for each tested approach. The relationships between stem curvature and the dating precision of the approaches were also analysed. The results of dating precision showed crucial differences between the tested approaches. Tree stem curvature did not influence dating precision. Overall, we concluded that the approaches based on eccentric increments are less precise than those based on reaction wood analysis when juvenile P. abies trees that grow on landslide slope are sampled. The importance of this study is in providing new insight into dating precision by analysing the whole tree stems of trees influenced by mass movements. This is unique since the approaches based on eccentricity increments have not been previously tested in such a detail.     

Developmental plasticity and biomechanics of treelets and lianas in Manihot aff. quinquepartita (Euphorbiaceae): a branch-angle climber of French Guiana

Background and Aims

Most tropical lianas have specialized organs of attachment such as twining stems, hooks or tendrils but some do not. Many climbers also have an early self-supporting phase of growth and in some species this can produce treelet-sized individuals. This study focuses on how a liana can climb without specialized attachment organs and how biomechanical properties of the stem are modulated between self-supporting treelets and canopy-climbing lianas.

Methods

Biomechanics and stem development were investigated in self-supporting to climbing individuals of Manihot aff. quinquepartita (Euphorbiaceae) from tropical rain forest at Saül, central French Guiana. Bending tests were carried out close to the site of growth. Mechanical properties, including Young''s elastic modulus, were observed with reference to habit type and changes in stem anatomy during development.

Key Results

This liana species can show a remarkably long phase of self-supporting growth as treelets with stiff, juvenile wood characterizing the branches and main stem. During the early phase of climbing, stiff but unstable stem segments are loosely held in a vertical position to host plants via petiole bases. The stiffest stems – those having the highest values of Young''s modulus measured in bending – belonged to young, leaning and climbing stems. Only when climbing stems are securely anchored into the surrounding vegetation by a system of wide-angled branches, does the plant develop highly flexible stem properties. As in many specialized lianas, the change in stiffness is linked to the development of wood with numerous large vessels and thin-walled fibres.

Conclusions

Some angiosperms can develop highly effective climbing behaviour and specialized flexible stems without highly specialized organs of attachment. This is linked to a high degree of developmental plasticity in early stages of growth. Young individuals in either open or closed marginal forest conditions can grow as substantial treelets or as leaning/climbing plants, depending on the availability of host supports. The species of liana studied differs both in terms of development and biomechanics from many other lianas that climb via twining, tendrils or other specialized attachment organs.Key words: Biomechanics, bending, developmental plasticity, French Guiana, liana, Manihot aff. quinquepartita (Euphorbiaceae), treelet, branch angle climber, Young''s modulus     

DEVELOPMENTAL ANATOMY OF JUVENILE AND ADULT SHOOTS OF MARCGRAVIA RECTIFOLIA L

Marcgravia rectifolia L. is a dimorphic vine having distinct juvenile and adult shoots. The juvenile shoot is a climber characterized by an orthotropic growth habit, a flattened stem, adventitious roots, and ovate leaves. The adult shoot, on the other hand, possesses a plagiotropic growth habit, has a cylindrical stem, few or no adventitious roots, and lanceolate leaves. Both phases have distichous phyllotaxy, however the plastochron is shorter for the adult phase than for the juvenile. Internode elongation occurs earlier for adult shoots than for juvenile shoots. Cytological analyses show the flattened stem of the juvenile results from differential production of cells, especially in the pith region. On the other hand, internodes of the adult phase are longer than juvenile internodes, a result of more cells produced rather than longer cells. In juvenile stems a perivascular band of elongated fibers develops, while in adult stems this band consists of brachyosclereids. Both phases undergo secondary growth and have non-storied cambia. Cambial activity begins in the 6th internode of each phase. As secondary growth proceeds, the adult stem produces much more xylem than juvenile stems of the same age. Adventitious roots produced in the juvenile stem are located in vertical rows at the “corners” of flattened stems and are attachment structures aiding the climbing habit of this vine. Phase changes occur regularly in this species. The juvenile phase usually transforms into the adult, however the adult phase can spontaneously revert back into the juvenile phase. The anatomical features and the phase changes are discussed and compared to Hedera helix, a vine whose phase changes have been studied in some detail. It is suggested that the anatomical features of Marcgravia rectifolia L. including its phase changes, may provide an alternative system to study physiological changes similar to those done with Hedera helix.     

Responses of two parasitoids,the exotic Spathius agrili Yang and the native Spathius floridanus Ashmead,to volatile cues associated with the emerald ash borer,Agrilus planipennis Fairmaire

Two parasitoids, the introduced specialist Spathius agrili Yang (Braconidae), and the native generalist Spathius floridanus Ashmead, have been proposed as biological control agents of the emerald ash borer, Agrilus planipennis Fairmaire (Buprestidae). However, little is known about their host-location behaviors. We evaluated responses to their host complex, Fraxinus pennsylvanica stem tissue, F. pennsylvanica foliage, and an A. planipennis larva within a stem. Experiments were conducted in a Y-tube olfactometer, using wasps reared on A. planipennis larvae in F. pennsylvanica stems. Naïve S. agrili were attracted to the entire complex, and to leaf tissue, relative to blanks. S. agrili were also more attracted to stems containing larvae and leaf tissue together than leaf tissue alone. Naïve S. floridanus were attracted to larvae within stems, but nothing else. A further distinction is that S. agrilli moved more, in the presence of foliage. Thus, S. agrili and S. floridanus appear to employ different host-location strategies. The former is attracted to host plant cues, which then elicit increased searching, whereas the latter is only attracted to infested tissue directly. We found no evidence that oviposition influences attraction by S. agrili, suggesting other forms of experience should be evaluated for potential sources of learned cues. Further, S. agrili that declined opportunities to oviposit oriented away from host-associated cues, suggesting distinct behavioral sequences occur by females that are not reproductively ready. Further understanding of host-location behavior may improve biological control by these parasitoids, by suggesting strategies for pre-release conditioning and providing tools for assessing post-release establishment.     

Trade-off between vegetative regeneration and sexual reproduction of <Emphasis Type="Italic">Sargassum thunbergii</Emphasis>

An ecological demographic study was conducted from January to December 2009 at Changshan Island on the northern side of Shandong Peninsula, China. The study investigated stem regeneration and the resource trade-off between sexual reproduction and vegetative regeneration of the brown alga Sargassum thunbergii at the individual thallus level. Stem regeneration from the holdfast occurred year round but with significant seasonality, peaking in the early summer. Both percentage of fertile stems and reproductive effort were negatively correlated with stem regeneration in the middle and later periods of sexual reproduction. Furthermore, the correlation coefficients increased significantly with time. These observations suggested that an increase in resource allocation to sexual reproduction would lead to a decrease in allocation to vegetative regeneration during the reproductive period. Reproductive effort was lower and stem regeneration was higher in the wave-exposed location compared with the sheltered location, indicating that allocation of resources between sexual reproduction and vegetative regeneration varied with location. When reproductive tissues were removed, the number of new stems produced from holdfasts increased, and when new stems were removed, an increase in reproductive effort occurred. These results suggest that trade-off between sexual reproduction and vegetative regeneration occurs in S. thunbergii.