Characterization of C₃--C₄ intermediate species in the genus Heliotropium L. (Boraginaceae): anatomy, ultrastructure and enzyme activity

Photosynthetic pathway characteristics were studied in nine species of Heliotropium (sensu lato, including Euploca), using assessments of leaf anatomy and ultrastructure, activities of PEP carboxylase and C₄ acid decarboxylases, and immunolocalization of ribulose 1·5‐bisphosphate carboxylase/oxygenase (Rubisco) and the P‐subunit of glycine decarboxylase (GDC). Heliotropium europaeum, Heliotropium calcicola and Heliotropium tenellum are C₃ plants, while Heliotropium texanum and Heliotropium polyphyllum are C₄ species. Heliotropium procumbens and Heliotropium karwinskyi are functionally C₃, but exhibit ‘proto‐Kranz’ anatomy where bundle sheath (BS) cells are enlarged and mitochondria primarily occur along the centripetal (inner) wall of the BS cells; GDC is present throughout the leaf. Heliotropium convolvulaceum and Heliotropium greggii are C₃–C₄ intermediates, with Kranz‐like enlargement of the BS cells, localization of mitochondria along the inner BS wall and a loss of GDC in the mesophyll (M) tissue. These C₃–C₄ species of Heliotropium probably shuttle photorespiratory glycine from the M to the BS tissue for decarboxylation. Heliotropium represents an important new model for studying C₄ evolution. Where existing models such as Flaveria emphasize diversification of C₃–C₄ intermediates, Heliotropium has numerous C₃ species expressing proto‐Kranz traits that could represent a critical initial phase in the evolutionary origin of C₄ photosynthesis.     

Pteronia divaricata (Asteraceae): A newly recorded Cape herbal medicine

The first ethnobotanical data on Pteronia divaricata is presented, which shows that the plant is an important traditional Cape medicine, especially in the Cederberg region (Western Cape Province) and in the Middelpos and Nieuwoudtville districts of the Northern Cape Province. Surprisingly, not a single publication could be found that records any uses for the species. We accurately documented nine separate anecdotes, which include various medicinal uses (for the treatment of colds, fever, influenza, stomach pain, diarrhoea, back pain, chest ailments, high blood pressure and tuberculosis). The novel data also includes seven previously unrecorded vernacular names, namely flip-se-bos, inflammasiebos, pylbos, dassiebos, dassiepisbos, perdebos and boegabos. Since the species is poorly known, its general morphology, leaf anatomy and essential oil composition were studied. Oil is produced in secretory ducts along the midribs below the main vascular bundle; secretory trichomes are also present on the leaf surface. The oil is relatively complex and contains a combination of sabinene, myrcene, β-caryophyllene and bicyclogermacrene as main compounds with smaller amounts of limonene, p-cymene, tetradecane, pentadecane, terpinen-4-ol and δ-cadinene. Dichloromethane extracts exhibited antibacterial activity (especially against Bacillus cereus) at MIC values as low as 1.0 mg/ml. Other solvent extracts and the essential oils were less active.     

Pericarp development and fruit structure in borassoid palms (Arecaceae-Coryphoideae-Borasseae)

Background and Aims

The Borasseae form a highly supported monophyletic clade in the Arecaceae–Coryphoideae. The fruits of Coryphoideae are small, drupaceous with specialized anatomical structure of the pericarp and berries. The large fruits of borassoid palms contain massive pyrenes, which develop from the middle zone of the mesocarp. The pericarp structure and mode of its development in Borasseae are similar to those of Eugeissona and Nypa. A developmental carpological study of borassoid palms will allow us to describe the process of pericarp development and reveal the diagnostic fruit features of borassoid palms, determine the morphogenetic fruit type in Borasseae genera, and describe similarities in fruit structure and pericarp development with other groups of palms.

Methods

The pericarp anatomy was studied during development with light microscopy based on the anatomical sections of fruits of all eight Borasseae genera.

Key Results

The following general features of pericarp structure in Borasseae were revealed: (1) differentiation of the pericarp starts at early developmental stages; (2) the exocarp is represented by a specialized epidermis; (3) the mesocarp is extremely multilayered and is differentiated into several topographical zones – a peripheral parenchymatous zone(s) with scattered sclerenchymatous elements and vascular bundles, a middle zone (the stony pyrene comprising networks of elongated sclereids and vascular bundles) and an inner parenchymatous zone(s); (4) differentiation and growth of the pyrene tissue starts at early developmental stages and ends long before maturation of the seed; (5) the inner parenchymatous zone(s) of the mesocarp is dramatically compressed by the mature seed; (6) the endocarp (unspecialized epidermis) is not involved in pyrene formation; and (7) the spermoderm is multilayered in Hyphaeninae and obliterated in Lataniinae.

Conclusions

The fruits of Borasseae are pyrenaria of Latania-type. This type of pericarp differentiation is also found only in Eugeissona and Nypa. The fruits of other Coryphoideae dramatically differ from Borasseae by the pericarp anatomical structure and the mode of its development.     

Comparative anatomy of the nectary spur in selected species of Aeridinae (Orchidaceae)

Background and Aims

To date, the structure of the nectary spur of Aeridinae has not been studied in detail, and data relating to the nectaries of ornithophilous orchids remain scarce. The present paper compares the structural organization of the floral nectary in a range of Aeridinae species, including both entomophilous and ornithophilous taxa.

Methods

Nectary spurs of Ascocentrum ampullaceum (Roxb.) Schltr. var. aurantiacum Pradhan, A. curvifolium (Lindl.) Schltr., A. garayi Christenson, Papilionanthe vandarum (Rchb.f.) Garay, Schoenorchis gemmata (Lindl.) J.J. Sm., Sedirea japonica (Rchb.f.) Garay & H.R. Sweet and Stereochilus dalatensis (Guillaumin) Garay were examined by means of light microscopy, scanning electron microscopy and transmission electron microscopy.

Key Results and Conclusions

The diverse anatomy of the nectary is described for a range of Aeridinae species. All species of Ascocentrum investigated displayed features characteristic of ornithophilous taxa. They have weakly zygomorphic, scentless, red or orange flowers, display diurnal anthesis, possess cryptic anther caps and produce nectar that is secluded in a relatively massive nectary spur. Unicellular, secretory hairs line the lumen at the middle part of the spur. Generally, however, with the exception of Papilionanthe vandarum, the nectary spurs of all entomophilous species studied here (Schoenorchis gemmata, Sedirea japonica, Stereochilus dalatensis) lack secretory trichomes. Moreover, collenchymatous secretory tissue, present only in the nectary spur of Asiatic Ascocentrum species, closely resembles that found in nectaries of certain Neotropical species that are hummingbird-pollinated and assigned to subtribes Maxillariinae Benth., Laeliinae Benth. and Oncidiinae Benth. This similarity in anatomical organization of the nectary, regardless of geographical distribution and phylogeny, indicates convergence.     

Comparative root anatomy of papilionoid Legumes

The aims of the present study were (i) to compare the anatomical structures of roots from legume species belonging to the tribes Aeschynomeneae, Fabeae (syn. Vicieae), and Trifolieae (Papilionoideae); (ii) to characterize those developmental stages of roots that are most useful for comparative studies; (iii) to characterize root developmental patterns during their growth from primary to adult structures; and (iv) to identify developmental patterns that are typical for particular tribes within this plant group. To this end, serial transverse cuts of the main roots of 20 species belonging to the above-mentioned tribes were made from the root tip to the root collar. Vessel density, vessel diameter, secondary cortex area, and xylem area were quantified using appropriate software. Based on root ontogeny, four stages giving evidence for important developmental events were chosen based on which a comparison of the species-specific structures became possible. The first stage corresponds to the primary structure of a root. The second stage corresponds to the onset of cambium and phellogen division, when their first division products become differentiated. The third and fourth stages correspond to adult roots with a well-developed secondary structure. These developmental stages that allowed between-species comparison turned out to be useful characterising those patterns that best typify a particular tribe within the subfamily. Vessel density, vessel diameter, and the xylem transverse area were found to differ significantly between the tribes examined. Some of the analyzed morphological variables had a tribe-specific consistency, so that, together with other characteristics, they should be included in approaches to clarify systematic problems in the legume family.     

Vertebral and rib anatomy in Caperea marginata: Implications for evolutionary patterning of the mammalian vertebral column

The pygmy right whale, Caperea marginata, is a rare mysticete cetacean with an unusual suite of axial skeletal characters. Distally expanded first ribs, a long thorax with broadly overlapping vertebral transverse processes, plate‐like posterior ribs, and a short tail contrast with other cetaceans and suggest unique developmental patterning. Twenty‐four individuals of diverse ontogenetic age were available for analysis. Multiple, variable examples of incomplete rib fusion in dependent calves indicate that the first rib of adults is an ontogenetic fusion product of ribs 1 and 2. The composite rib articulates by way of its anterior (Rib1) component to the sternum and by way of its posterior (Rib2) component with thoracic vertebra 2. Thoracic vertebra 1 lacks rib articulations. When rib fusion is taken into account, the most frequent column formulas are C7T18L1Cd16–17 = 42–43 and C7T17L1Cd16–18 = 41–43. Thoracic and lumbar series are not reciprocal in count, arguing against their developmental linkage. Instead, parallel reduction in both lumbar and caudal counts supports the existence of neocete patterning in Caperea, as in all other living cetaceans. Ontogenetic vertebral column elongation is most marked in the posterior thorax, lumbos, and anterior tail. Vertebrae in these column regions are excellent predictors of total body length.     

The phylogenetic distribution of the ampulla ureter and ampulla urogenital/uriniferous papilla in the Serpentes

The ampulla ureter and ampulla urogenital/uriniferous papilla represent differing morphologies of the caudal urogenital ducts in snakes. The ampulla ureter is an enlarged portion of the caudal extremity of the ureter that communicates the cranial regions of the ureter and the ductus deferens/Wolffian duct to the urodaeum. The ampulla urogenital/uriniferous papilla is an enlarged pouch, distinct from the ureter, which communicates the ureter and ductus deferens/Wolffian duct to the urodaeum. Although functional differences of these two structures are unknown, the ampulla urogenital/uriniferous papilla may have evolved for urine storage in males and females, and secondarily evolved a reproductive function in males. The most parsimonious optimization of the ampulla ureter and ampulla urogenital/uriniferous papilla indicates that the ampulla ureter is the ancestral state in snakes. Examining the presence or absence of the ampulla ureter and ampulla urogenital/uriniferous papilla in snakes on conflicting caenophidian phylogenies results in two hypotheses for the evolution of these variant morphologies: (1) The ampulla urogenital/uriniferous papilla evolved from the ampulla ureter independently in the Colubroidea and Elapoidea with subsequent losses of the ampulla urogenital/uriniferous papilla in the Elapoidea and (2) a single transition from the ampulla ureter to the ampulla urogenital/uriniferous papilla on the branch leading to the Colubroidea + Elapoidea with subsequent losses of the ampulla urogenital/uriniferous papilla in the Elapoidea and Colubroidea. The presence of the ampullae urogenital/uriniferous papilla in only the Colubroidea and Elapoidea highlights the affinity of these two taxonomic groups, a relationship that is strongly supported in published cladograms produced with molecular datasets.     

Stem tissue mass density is linked to growth and resistance to a stem‐boring insect in Alternanthera philoxeroides

To investigate how stem anatomical structure is linked to growth and resistance to stem‐boring insects in a herbaceous species, six populations of alligatorweed (Alternanthera philoxeroides) were grown in a common garden. Stem growth rate (GR) of A. philoxeroides and pupation rate as an estimate of resistance to a stem‐boring insect (Agasicles hygrophila) were quantified. Stem tissue mass density (TMD) was measured and stem anatomical traits were analysed on cross‐sectional areas (CSA). Stem TMD was positively correlated with resistance (i.e. negatively correlated with pupation rate) and negatively correlated with GR. Stem cortex CSA (%) and vascular bundle (VB) density (no./mm²) were positively related to stem TMD and negatively related to pupation rate. The GR was positively related to VB CSA (%) and negatively related to VB density. These results suggest that stem TMD, which results from a high fraction in cortex CSA and high VB density, is a key determinant of resistance to a stem‐boring specialist in A. philoxeroides. The high resistance of plants with higher stem TMD may partially impose a cost to plant growth.     

Response of rigor cross-bridges to stretch detected by fluorescence lifetime imaging microscopy of myosin essential light chain in skeletal muscle fibers

We applied fluorescence lifetime imaging microscopy to map the microenvironment of the myosin essential light chain (ELC) in permeabilized skeletal muscle fibers. Four ELC mutants containing a single cysteine residue at different positions in the C-terminal half of the protein (ELC-127, ELC-142, ELC-160, and ELC-180) were generated by site-directed mutagenesis, labeled with 7-diethylamino-3-((((2-iodoacetamido)ethyl)amino)carbonyl)coumarin, and introduced into permeabilized rabbit psoas fibers. Binding to the myosin heavy chain was associated with a large conformational change in the ELC. When the fibers were moved from relaxation to rigor, the fluorescence lifetime increased for all label positions. However, when 1% stretch was applied to the rigor fibers, the lifetime decreased for ELC-127 and ELC-180 but did not change for ELC-142 and ELC-160. The differential change of fluorescence lifetime demonstrates the shift in position of the C-terminal domain of ELC with respect to the heavy chain and reveals specific locations in the lever arm region sensitive to the mechanical strain propagating from the actin-binding site to the lever arm.     

Fibroblast growth factor receptor 3 (FGFR3) is a strong heat shock protein 90 (Hsp90) client: implications for therapeutic manipulation

Fibroblast growth factor receptor 3 (FGFR3) is a key regulator of growth and differentiation, whose aberrant activation causes a number of genetic diseases including achondroplasia and cancer. Hsp90 is a specialized molecular chaperone involved in stabilizing a select set of proteins termed clients. Here, we delineate the relationship of Hsp90 and co-chaperone Cdc37 with FGFR3 and the FGFR family. FGFR3 strongly associates with these chaperone complexes and depends on them for stability and function. Inhibition of Hsp90 function using the geldanamycin analog 17-AAG induces the ubiquitination and degradation of FGFR3 and reduces the signaling capacity of FGFR3. Other FGFRs weakly interact with these chaperones and are differentially influenced by Hsp90 inhibition. The Hsp90-related ubiquitin ligase CHIP is able to interact and destabilize FGFR3. Our results establish FGFR3 as a strong Hsp90 client and suggest that modulating Hsp90 chaperone complexes may beneficially influence the stability and function of FGFR3 in disease.