Natural foliar variegation without costs? The case of Begonia

Background and Aims

Foliar variegation is recognized as arising from two major mechanisms: leaf structure and pigment-related variegation. Begonia has species with a variety of natural foliar variegation patterns, providing diverse examples of this phenomenon. The aims of this work are to elucidate the mechanisms underlying different foliar variegation patterns in Begonia and to determine their physiological consequences.

Methods

Six species and one cultivar of Begonia were investigated. Light and electron microscopy revealed the leaf structure and ultrastructure of chloroplasts in green and light areas of variegated leaves. Maximum quantum yields of photosystem II were measured by chlorophyll fluorescence. Comparison with a cultivar of Ficus revealed key features distinguishing variegation mechanisms.

Key Results

Intercellular space above the chlorenchyma is the mechanism of variegation in these Begonia. This intercellular space can be located (a) below the adaxial epidermis or (b) below the adaxial water storage tissue (the first report for any taxa), creating light areas on a leaf. In addition, chlorenchyma cell shape and chloroplast distribution within chlorenchyma cells differ between light and green areas. Chloroplasts from both areas showed dense stacking of grana and stroma thylakoid membranes. The maximum quantum yield did not differ significantly between these areas, suggesting minimal loss of function with variegation. However, the absence of chloroplasts in light areas of leaves in the Ficus cultivar led to an extremely low quantum yield.

Conclusions

Variegation in these Begonia is structural, where light areas are created by internal reflection between air spaces and cells in a leaf. Two forms of air space structural variegation occur, distinguished by the location of the air spaces. Both forms may have a common origin in development where dermal tissue becomes loosely connected to mesophyll. Photosynthetic functioning is retained in light areas, and these areas do not include primary veins, potentially limiting the costs of variegation.     

Leaf micromorphology and leaf glandular hair ontogeny of Myoporum bontioides A. Gray

Myoporum bontioides A. Gray (Myoporaceae), a red list plant in Japan, is restricted to only a few East Asian countries like China, Japan and Taiwan, associated to some true mangroves. The leaf is isolateral and has a thick cuticle; stomata are anomocytic, sunken and have a beak‐shaped cuticular outgrowth at the inner and outer side of the stomatal pore (ledges); profuse glandular hairs are scattered on both leaf surfaces of young leaves. The mesophyll is compact with palisade and spongy parenchyma cells in the young stage, but at maturity profuse intercellular spaces can be observed. Secretory ducts occur in young leaves. Pear‐shaped glandular hairs protrude from the epidermal layer. Hair primordia are well distinct by their larger size and undergo divisions to produce two laterally placed basal cells, two stalk cells and four radiating terminal cells. The cuticle layers of the terminal cells are often separated from the cell wall to form a space, in which ions accumulate for excretion. Inner walls of the basal cells are connected with the mesophyll. Though ontogeny and structure of glandular hairs have resemblance to typical mangroves, considering leaf micromorphology, this plant is better termed as “mangrove associate” instead of “true mangrove”. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Multiple canopy opening effects on recruited saplings in a typhoon-disturbed tropical rainforest,Taiwan

To investigate the influence of multiple canopy openings on the composition and diversity of recruited saplings in a forest frequently disturbed by typhoons. We conducted tree-by-tree censuses (diameter at breast height ≥ 1 cm) and mapped gaps (canopy height < 5 m) in 1993, 2000, 2008, and 2013 in a tropical mountain zonal foothill evergreen broad-leaved forest in Nanjenshan Nature Reserve, Taiwan. We analyzed the composition and diversity of recruited saplings within a 2.1 ha plot (840 sampling quadrats (5 m × 5 m)) with variable numbers of canopy openings recorded during the study period. Composition of recruited saplings was dissimilar between quadrats that stayed opened and those that stayed closed throughout the study period (pairwise similarity estimates C₀₂ = 0.52, 95% CI = 0.38–0.66). The quadrats under closed canopy had high diversity when weighting rare species (species richness), whereas quadrats with one or two gap opening records during the past 20 years had high diversity when weighting the abundance of species. Although canopy openings provided establishment conditions for saplings of some shade-intolerant species, due to the nature of small gap size, such habitats do not favor the dominance of shade-intolerant species. Even in a frequently disturbed forest, species composition and richness of recruited saplings were mainly contributed by shade-tolerant species. Although multiple canopy openings facilitated the establishment of shade-intolerant species, species diversity in the study forests is possibly mainly mediated by coexistence mechanisms of those shade-tolerant species rather than light-gap-related species strategies.     

A new classification of marginal resin ducts improves understanding of hard pine (Pinaceae) diversity in Taiwan

Resin ducts provide important characters for classifying the Pinaceae. Here we study Pinus massoniana and P. taiwanensis and show that the generally-used term marginal (=external) resin duct, applied to ducts in needle leaves, needs to be further differentiated into marginal (strongly attaching to the dermal tissue, and lacking the complete ring structure formed by the sheath cells) and submarginal ducts (adjacent to hypodermal cells, with a complete ring structure formed by the sheath cells). On this basis P. massoniana and P. taiwanensis, which are nearly indistinguishable based on external morphology, are clearly differentiated. Their similar morphology has led to a long standing debate on where P. massoniana occurs in Taiwan. Based on this new classification of resin ducts, we examined old herbarium specimens previously identified as P. massoniana, surveyed current hard pines in Taiwan, and checked historical documents. Needle leaves of these two taxa were studied and compared with the material of P. massoniana from mainland China as a reference. Pinus massoniana shows almost exclusively marginal resin ducts, with extremely rare changes along the length of the duct to submarginal positions. In contrast, P. taiwanensis shows a mixture of medial, submarginal, septal, and rarely endonal ducts, with occasional changes of the duct between various types. Re-identification of specimens showed that only 5 specimens are possible native P. massoniana collected from northern Taiwan and the others are all P. taiwanensis. The hard pine of Junjianyan is unexpectedly found to be the only currently known P. massoniana in Taiwan, which is likely a relic of historical afforestation. The hard pine from Huoyansan and the Coastal Range, which is widely accepted as P. massoniana, is not this species. In addition to the taxonomic value of the new definition applied to these two studied taxa, we expect that this approach can generally be applied distinguishing the respective characters in the genus Pinus and in other conifers.     

Phylogenetics and Biogeography of the <Emphasis Type="Italic">Phalaenopsis violacea</Emphasis> (Orchidaceae) Species Complex Based on Nuclear and Plastid DNA

The Phalaenopsis violacea complex includes two species: P. violacea Witte and Phalaenopsis bellina (Rchb.f.) E. A. Christ. However, three forms of P. violacea have been found in different areas, including Sumatra, the Malay Peninsula, and Mentawai Island. The phylogenetic tree inferred from the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA), the trnL intron, and the atpB-rbcL spacer of plastid DNA were used to clarify the phylogenetics and biogeography of the P. violacea complex. Analyses of the trnL intron sequences and of the atpB-rbcL spacer did not allow for apparent discrimination among these three species of the P. violacea complex. Based on the phylogenetic tree inferred from the ITS sequence, P. bellina cannot be separated from populations of P. violacea, with the exception of the population distributed on Mentawai Is., Indonesia. Based on morphological characteristics, P. violacea distributed on Mentawai Is. has a long and roundish rachis and is separate from the other groups of the P. violacea complex described by Christenson (Timber, Portland, OR, 2001). Therefore, the results of this study show a trend that supports the conclusion that the population of the P. violacea complex on Mentawai Is. is a separate species from P. violacea. Based on the biogeography of the P. violacea complex, Mentawai plants of this complex might be descended from those on the Sumatra/Malay Peninsula.     

Bizonoplast, a unique chloroplast in the epidermal cells of microphylls in the shade plant Selaginella erythropus (Selaginellaceae)

Study of the unique leaf anatomy and chloroplast structure in shade-adapted plants will aid our understanding of how plants use light efficiently in low light environments. Unusual chloroplasts in terms of size and thylakoid membrane stacking have been described previously in several deep-shade plants. In this study, a single giant cup-shaped chloroplast, termed a bizonoplast, was found in the abaxial epidermal cells of the dorsal microphylls and the adaxial epidermal cells of the ventral microphylls in the deep-shade spike moss Selaginella erythropus. Bizonoplasts are dimorphic in ultrastructure: the upper zone is occupied by numerous layers of 2-4 stacked thylakoid membranes while the lower zone contains both unstacked stromal thylakoids and thylakoid lamellae stacked in normal grana structure oriented in different directions. In contrast, other cell types in the microphylls contain chloroplasts with typical structure. This unique chloroplast has not been reported from any other species. The enlargement of epidermal cells into funnel-shaped, photosynthetic cells coupled with specific localization of a large bizonoplast in the lower part of the cells and differential modification in ultrastructure within the chloroplast may allow the plant to better adapt to low light. Further experiments are required to determine whether this shade-adapted organism derives any evolutionary or ecophysiological fitness from these unique chloroplasts.     

Lamelloplasts and minichloroplasts in Begoniaceae: iridescence and photosynthetic functioning

Iridoplasts (modified plastids in adaxial epidermal cells) reported from Begonia were originally hypothesized to cause iridescence, which was broadly accepted for decades. However, several species of Begonia with iridoplasts are not iridescent causing confusion. Here chloroplast ultrastructure was observed in 40 taxa of Begoniaceae to explore the phenomenon of iridescence. However, 22 Begonias and Hillebrandia were found to have iridoplasts, but only nine display visually iridescent blue to blue-green leaves. Unexpectedly, a new type of plastid, a ‘minichloroplast,’ was found in the abaxial epidermal cells of all taxa, but was present in adaxial epidermal cells only if iridoplasts were absent. Comparative ultrastructural study of iridoplasts and a shading experiment of selected taxa show that a taxon with iridoplasts does not inevitably have visual iridescence, but iridescence is greatly affected by the spacing between thylakoid lamellae (stoma spacing). Thus, we propose instead the name ‘lamelloplast’ for plastids filled entirely with regular lamellae to avoid prejudging their function. To evaluate photosynthetic performance, chlorophyll fluorescence (F _v /F _m ) was measured separately from the chloroplasts in the adaxial epidermis and lower leaf tissues by using leaf dermal peels. Lamelloplasts and minichloroplasts have much lower photosynthetic efficiency than mesophyll chloroplasts. Nevertheless, photosynthetic proteins (psbA protein of PSII, RuBisCo and ATPase) were detected in both plastids as well as mesophyll chloroplasts in an immunogold labeling. Spectrometry revealed additional blue to blue-green peaks in visually iridescent leaves. Micro-spectrometry detected a blue peak from single blue spots in adaxial epidermal cells confirming that the color is derived from lamelloplasts. Presence of lamelloplasts or minichloroplasts is species specific and exclusive. High prevalence of lamelloplasts in Begoniaceae, including the basal clade Hillebrandia, highlights a unique evolutionary development. These new findings clarify the association between iridescence and lamelloplasts, and with implications for new directions in the study of plastid morphogenesis.     

Simple liquid chromatographic method for the determination of cefotaxime in human and rat plasma

A high-performance liquid chromatographic method with ultraviolet (UV) detection was developed for measuring cefotaxime in rat and human plasma. The method used direct injection of the plasma supernatant after deproteinization with 70% perchloric acid. Degradation of cefotaxime in acidic medium was retarded by adding phosphate buffer before centrifuging the sample. The mobile phase was 0.05 M aqueous ammonium acetate-acetonitrile-tetrahydrofuran (87:11:2, v/v) adjusted to pH 5.5. Analysis was run at a flow-rate of 1.0 ml/min, and a detection wavelength of 254 nm was used. The method has a quantification limit of 0.20 microgram/ml. The within- and between-day coefficients of variation and accuracy values were less than 8% and +/-3%, respectively, while the recovery values were greater than 87% over the concentration range tested (0.20-50 microgram/ml). The speed, sensitivity, specificity and reproducibility of this method make it particularly suitable for the routine determination of cefotaxime in human plasma. Moreover, only a relatively small sample plasma volume (100 microliter) is required, allowing this method to be applied to samples taken from neonates.     

Comparative systematic study of colleters and stipules of Rhizophoraceae with implications for adaptation to challenging environments

Colleters are multicellular secretory structures found on various organs in flowering plants. Colleters on the adaxial sides of stipules have been hypothesized to play a role in protecting the developing shoot. Rhizophoraceae is a stipulate family with a broad distribution from mangrove to montane environments, which makes the family well suited for the examination of this hypothesis, but the colleters of Rhizophoraceae are not well known. We compared species from all three tribes of Rhizophoraceae, including five inland genera and all four mangrove genera. In all species, several to hundreds of colleters, sessile or stalked, arranged in rows aggregated in genus‐specific shapes, are found at the adaxial bases of open and closed stipules. Pellacalyx uniquely has additional colleters at the stipule margins. Colleters are all of the standard type, comprising a central axis of core parenchyma with large vacuoles and tannins, and an outer palisade‐like epidermis with organelles involved in secretory activity. An exception is Pellacalyx axillaris, in which colleters appear as extremely small epidermal protrusions. Kandelia obovata has a tracheary element in some colleters. Pellacalyx uniquely has an unusual fleshy outgrowth on the adaxial stipule base. We propose an evolutionary sequence in which Macarisia has plesiomorphic stipule and colleter traits and the mangrove Kandelia obovata with colleter vascular traces is most derived. Colleter and stipule structures are largely concordant with habitat and phylogeny, and show taxonomic value. The strong alignment of colleter and stipule patterns with habitat is suggestive that colleters have a protective function, although some components of these patterns may be phylogenetically determined. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 449–464.