Scaling relationships and vessel packing in petioles.

PREMISE OF THE STUDY: While tradeoffs among mechanical and conductive functions have been well investigated in woody stems, these tradeoffs are relatively unexplored in petioles, the structural link between stems and laminas. We investigated size-independent scaling relationships between cross-sectional areas of structural and vascular tissues, relationships between tissue areas of xylem and phloem, vessel packing within xylem, and scaling of vascular and structural tissues with lamina traits. METHODS: We examined allometric relationships among petiole tissues and as a function of lamina and petiole size variation on eleven species of Pelargonium. From transverse sections of methacrylate-embedded tissue, we measured the cross-sectional areas of all tissues within the petiole and vessel lumen, and cell wall areas of each vessel. Allometric scaling relationships were analyzed using standardized major axis regressions. KEY RESULTS: Pelargonium petiole vessels were packed as predicted by Sperry's packing rule for woody stems. In contrast to woody stems, there was no evidence of a tradeoff between vessel area and fiber area. Within cross-sections, more xylem was produced than phloem. Among bundles, xylem and phloem scaling relationships varied with bundle position. Except for lamina dry mass and petiole fiber cross-sectional area, petiole and lamina traits were independent. CONCLUSIONS: Petioles share vascular tissue traits with stems despite derivation from leaf primordia. We did not find evidence for a tradeoff between structural and vascular tissues, in part because fibers occur outside the xylem. We propose this separation of conduction and support underlies observed developmental and evolutionary plasticity in petioles.

Contrasting lengths of Pelargonium floral nectar tubes result from late differences in rate and duration of growth.

Background and Aims: Much of morphological evolution in flowers has arisen from pollinator-mediated selection, often manifest as a match between the length of the pollinator's proboscis and the depth of tubular corollas or spurs. We investigate development, growth and homology of the unique nectar tube of Pelargonium, frequently described as 'a spur adnate to the pedicel'. Methods: We focused on two species. The nectar tube of P. ionidiflorum is three times longer than that of P. odoratissimum. Light and scanning electron microscopy were carried out, and daily growth measurements were used to compare nectar tube development and vascular patterns. Key Results: Nectar tubes in both species are initiated centripetally to the dorsal sepal in a space created by lateral displacement of two antepetalous stamens. The cavity deepens through subsequent intercalary growth of the receptacle that proceeds at the same rate in both species until tubes reach approx. 10 mm in length. Differences in final nectar tube lengths arise via an increase in the rate and duration of growth of the receptacle that begins just before anthesis (floral opening) and continues for several days past anthesis in P. ionidiflorum but does not occur in P. odoratissimum. Epidermal cells of the dorsal surface of the nectar tube in P. ionidiflorum are approx. 1.6 times longer than those in P. odoratissimum. Histological sections show no evidence that the nectar tube is a spur that became evolutionarily fused to the pedicel. Conclusions: Nectar tubes in Pelargonium are localized cavities that form in the receptacle via intercalary growth. Differences in the rate and duration of growth just prior to and following anthesis underlie differences in final tube lengths. Because differences in cell lengths do not fully account for differences in nectar tube lengths, evolutionary diversification must involve changes in both cell cycle and cell expansion.

Impact of plant growth promoting Pseudomonas monteilii PsF84 and Pseudomonas plecoglossicida PsF610 on metal uptake and production of secondary metabolite (monoterpenes) by rose-scented geranium (Pelargonium graveolens cv. bourbon) grown on tannery sludge amended soil.

Bacterial strains PsF84 and PsF610 were isolated from tannery sludge polluted soil, Jajmau, Kanpur, India. 16S rRNA gene sequence and phylogenetic analysis confirmed the taxonomic affiliation of PsF84 as Pseudomonas monteilii and PsF610 as Pseudomonas plecoglossicida. A greenhouse study was carried out with rose-scented geranium (Pelargonium graveolenscv. bourbon) grown in soil treated with tannery sludge in different proportions viz. soil: sludge ratio of 100:0, 25:75, 50:50, 75:25 and 0:100 to evaluate the effects of bacterial inoculation on the heavy metal uptake. The isolates solubilized inorganic phosphorus and were capable of producing indole acetic acid (IAA) and siderophore. The isolate PsF84 increased the dry biomass of shoot by 44%, root by 48%, essential oil yield 43% and chlorophyll by 31% respectively over uninoculated control. The corresponding increase with the isolate PsF610 were 38%, 40%, 39% and 28%, respectively. Scanning electron microscopic (SEM) studies reveal that the Cr(VI) accumulation resulted in breakdown of vascular bundles and sequesters Cr(VI) in roots. The glandular trichomes (GT) were investigated using SEM studies as these glands are probably the main site of essential oil synthesis. Owing to its wide action spectrum, these isolates could serve as an effective metal sequestering and bioinoculants due to the production of IAA, siderophore and solubilization of phosphate for geranium in metal-stressed soil. The present study has provided a new insight into the phytoremediation of metal-contaminated soil.

Phylogenetic influences on leaf trait integration in Pelargonium (Geraniaceae): convergence, divergence, and historical adaptation to a rapidly changing climate.

PREMISE OF THE STUDY: Trait integration may improve prediction of species and lineage responses to future climate change more than individual traits alone, particularly when analyses incorporate effects of phylogenetic relationships. The South African genus Pelargonium contains divergent major clades that have radiated along the same seasonal aridity gradient, presenting the opportunity to ask whether patterns of evolution in mean leaf trait values are achieved through the same set of coordinated changes among traits in each clade. METHODS: Seven leaf traits were measured on field-collected leaves from one-third of the species (98) of the genus. Trait relationships were examined using phylogenetic regression within major clades. Disparity analysis determined whether the course of trait evolution paralleled historical climate change events. KEY RESULTS: Divergence in mean trait values between sister clades A1 and A2 was consistent with expectations for leaves differing in longevity, despite strong similarity between clades in trait interactions. No traits in either clade exhibited significant relationships with multivariate climate axes, with one exception. Species in clades C and A2 included in this study occupied similar environments. These clades had similar values of individual trait means, except for δ(13)C, but they exhibited distinctive patterns of trait integration. CONCLUSIONS: Differing present-day patterns of trait integration are consistent with interpretations of adaptive responses to the prevailing climate at the time of each clade's origin. These differing patterns of integration are likely to exert strong effects on clade-level responses to future climate change in the winter rainfall region of South Africa.

Contributions regarding the leaf histo-anatomy of some Pelargonium species.

UNLABELLED: Pelargonium L. genra (Fam. Geraniaceae) includes over 250 species, most of it native to southern Africa. Nowadays, four perennial species are cultivated in Romania as ornamental plants. Our aim was to establish the main characteristics and differences that occur between Pelargonium zonale, P. hispidum, P. grandiflorum, P. peltatum and P. radens. All in all, the quantitative differences (regarding the leaf-stalk, blade and trichomes) have a taxonomical value for all five species included in the research. MATERIALS AND METHODS: The plant material was represented by fresh leaves harvested from 5 species of Pelargonium: P. peltatum (L.) Aiton, P. radens H.E. Moore, P. grandiflorum (Andrz.) Willd., P. hispidum. (anthesis phase) and preserved in ethanol 70%. Using a dissecting microscope, sections were made through leaf blade surface and cross sections of/ for leaf stalk and blade. RESULTS: The common characteristics of the leaf is the palmatilobate blade with long petiole. Although many studies regarded glandular trichomes from Pelargonium leaves, most of the leaf structures are still unknown. The most important for the quality and quantity of essential oil extracted from Pelargonium sp. are trichoms.

Comparison of leaf morphology and anatomy among Malva sylvestris ("gerânio-aromático"), Pelargonium graveolens ("falsa-malva") and Pelargonium odoratissimum ("gerânio-de-cheiro") / Comparação da Morfologia e da anatomia foliar entre as espécies Malva sylvestris (gerânio-aromático), Pelargonium graveolens (falsa-malva) e Pelargonium odoratissimum (gerânio-de-cheiro)

Malva sylvestris is generally confused with Pelargonium graveolens and Pelargonium odoratissimum due to similarities in their leaf morphology. The leaves of M. sylvestris have anthocyanins with scientifically proven cytotoxic, anti-inflammatory, antitumor and antioxidant properties. The leaves of P. odoratissimum have essential oil with antibacterial and spasmolytic properties, while the essential oil from P. graveolens has antimicrobial and antifungal activity. The aim of this study was to morpho-anatomically analyze the leaves of these species, indicating differences that can be used to clarify controversies about their use as medicinal plants. To anatomically compare the structure of each plant, samples were observed by Light Microscopy and Scanning Electron Microscopy (SEM). Leaf anatomy among species was quite different. Malva sylvestris showed capitate starry tector trichomes, as well as druses and mucilaginous cells. P. graveolens and P. odoratissimum differed as to trichomes. Both species had tector and glandular trichomes, and P. graveolens is distinguished for the greater height of tector trichomes and less quantity of the latter relative to P. odoratissimum. This study allowed the detection of anatomical differences, assisting in the taxonomy and classification of these species.

Malva sylvestris é comumente confundida com Pelargonium graveolens e Pelargonium odoratissimum devido às semelhanças na morfologia foliar. As folhas de M. sylvestris possuem antocianinas com propriedades citotóxicas, antiinflamatória, antitumoral e antioxidante já comprovadas cientificamente. As folhas de P. odoratissimum apresentam óleo essencial com propriedades antibacteriana e espasmolítica, e o óleo essencial da folha de P. graveolens possui atividade antimicrobiana e antifúngica. O objetivo deste estudo foi analisar morfo-anatomicamente as folhas destas espécies, apontando diferenças que possam ser utilizadas para esclarecer controvérsias na sua utilização como planta medicinal. Com a finalidade de se comparar anatomicamente a estrutura de cada planta, as amostras foram observadas por Microscopia de Luz e Microscopia Eletrônica de Varredura (MEV). A anatomia foliar entre as espécies foi bem distinta. Malva sylvestris apresentou tricomas do tipo capitado, estrelado e tector, além de drusas e células mucilaginosas. A distinção entre P. graveolens e P. odoratissimum foi observada em relação aos tricomas. Ambas as espécies apresentaram tricomas glandulares e tectores, sendo que P. graveolens se diferencia pela maior altura dos tricomas tectores e menor quantidade destes em relação ao P. odoratissimum. Este trabalho permitiu constatar diferenças anatômicas, auxiliando na taxonomia e classificação entre estas espécies.

Casparian bands occur in the periderm of Pelargonium hortorum stem and root.

BACKGROUND AND AIMS: Casparian bands are characteristic of the endodermis and exodermis of roots, but also occur infrequently in other plant organs, for example stems and leaves. To date, these structures have not been detected in phellem cells of a periderm. The aim of this study was to determine whether Casparian bands occur in phellem cells using tests that are known to detect Casparian bands in cells that also contain suberin lamellae. Both natural periderm and wound-induced structures were examined in shoots and roots. METHODS: Using Pelargonium hortorum as a candidate species, the following tests were conducted: (1) staining with berberine and counterstaining with aniline blue, (2) mounting sections in concentrated sulphuric acid and (3) investigating the permeability of the walls with berberine as an apoplastic, fluorescent tracer. KEY RESULTS: (1) Berberine-aniline blue staining revealed a modification in the radial and transverse walls of mature phellem cells in both stems and roots. Three days after wounding through to the cortex of stems, the boundary zone cells (pre-existing, living cells nearest the wound) had developed vividly stained primary walls. By 17 d, staining of mature phellem cells of wound-induced periderm was similar to that of natural periderm. (2) Mature native phellem cells of stems resisted acid digestion. (3) Berberine was excluded from the anticlinal (radial and transverse) walls of mature phellem cells in stems and roots, and from the wound-induced boundary zone. CONCLUSIONS: Casparian bands are present in mature phellem cells in both stems and roots of P. hortorum. It is proposed that Casparian bands act to retard water loss and pathogen entry through the primary cell walls of the phellem cells, thus contributing to the main functions of the periderm.

The 13C/12C isotopic signal of day-respired CO2 in variegated leaves of Pelargonium × hortorum.

In leaves, although it is accepted that CO(2) evolved by dark respiration after illumination is naturally (13) C-enriched compared to organic matter or substrate sucrose, much uncertainty remains on whether day respiration produces (13) C-depleted or (13) C-enriched CO(2). Here, we applied equations described previously for mesocosm CO(2) exchange to investigate the carbon isotope composition of CO(2) respired by autotrophic and heterotrophic tissues of Pelargonium × hortorum leaves, taking advantage of leaf variegation. Day-respired CO(2) was slightly (13) C-depleted compared to organic matter both under 21% O(2) and 2% O(2). Furthermore, most, if not all CO(2) molecules evolved in the light came from carbon atoms that had been fixed previously before the experiments, in both variegated and green leaves. We conclude that the usual definition of day respiratory fractionation, that assumes carbon fixed by current net photosynthesis is the respiratory substrate, is not valid in Pelargonium leaves under our conditions. In variegated leaves, total organic matter was slightly (13) C-depleted in white areas and so were most primary metabolites. This small isotopic difference between white and green areas probably came from the small contribution of photosynthetic CO(2) refixation and the specific nitrogen metabolism in white leaf areas.

Leaf shape evolution in the South African genus Pelargonium L'Her. (Geraniaceae).

Leaf shapes reflect complex assemblages of shape-determining elements, yet evolutionary studies tend to treat leaf shape as a single attribute, for example cordate or linear. As with all complex structures, individual elements of a leaf could theoretically evolve independently and at different rates to the extent permitted by genetic and functional limitations. We examined relative evolutionary lability of shape-determining elements in the highly diverse South African plant genus Pelargonium (Geraniaceae). We used SIMMAP to calculate Bayesian posterior probabilities for ancestral states of leaf-shape characters for major nodes across multiple phylogenetic trees. Trees were derived from a Bayesian analysis of DNA sequence data from four partitions. We found that shape elements differed in rates of character-state transformations across the tree. Leaf base, apex, and overall outline had low rates. Transformations in venation occurred at slightly higher rates and were associated with shifts in venation among major clades. Leaf margin type and overall leaf size showed intermediate rates, whereas high rates were observed in the extent of lamina lobing and functional leaf size. The results indicate that suites of elements characteristic of the recently evolved xerophytic lineage, for example pinnate venation, dissected lamina, and entire margins, were acquired piecemeal over nested levels of the phylogeny.

Leaf shape linked to photosynthetic rates and temperature optima in South African Pelargonium species.

The thermal response of gas exchange varies among plant species and with growth conditions. Plants from hot dry climates generally reach maximal photosynthetic rates at higher temperatures than species from temperate climates. Likewise, species in these environments are predicted to have small leaves with more-dissected shapes. We compared eight species of Pelargonium (Geraniaceae) selected as phylogenetically independent contrasts on leaf shape to determine whether: (1) the species showed plasticity in thermal response of gas exchange when grown under different water and temperature regimes, (2) there were differences among more- and less-dissected leafed species in trait means or plasticity, and (3) whether climatic variables were correlated with the responses. We found that a higher growth temperature led to higher optimal photosynthetic temperatures, at a cost to photosynthetic capacity. Optimal temperatures for photosynthesis were greater than the highest growth temperature regime. Stomatal conductance responded to growth water regime but not growth temperature, whereas transpiration increased and water use efficiency (WUE) decreased at the higher growth temperature. Strikingly, species with more-dissected leaves had higher rates of carbon gain and water loss for a given growth condition than those with less-dissected leaves. Species from lower latitudes and lower rainfall tended to have higher photosynthetic maxima and conductance, but leaf dissection did not correlate with climatic variables. Our results suggest that the combination of dissected leaves, higher photosynthetic rates, and relatively low WUE may have evolved as a strategy to optimize water delivery and carbon gain during short-lived periods of high soil moisture. Higher thermal optima, in conjunction with leaf dissection, may reflect selection pressure to protect photosynthetic machinery against excessive leaf temperatures when stomata close in response to water stress.

Geographic variation and plasticity to water and nutrients in Pelargonium australe.

Here, patterns of phenotypic plasticity and trait integration of leaf characteristics in six geographically discrete populations of the perennial herb Pelargonium australe were compared. It was hypothesized that populations would show local adaptation in trait means, but similar patterns of plasticity and trait integration. Further, it was questioned whether phenotypic plasticity was positively correlated with environmental heterogeneity and whether plasticity for water-use traits in particular was adaptive. Seedlings were grown in a glasshouse at six combinations of water and nutrient availability. Leaf anatomical, morphological and gas exchange traits were measured. High amounts of plasticity in leaf traits were found in response to changes in growth conditions and there was evidence of local adaptation among the populations. While there were significant correlations between plasticity and environmental heterogeneity, not all were positive. Notably, patterns of plasticity and trait integration varied significantly among populations. Despite that variation, some of the observed plasticity was adaptive: fitness was correlated with conservative water use when water was limiting. Pelargonium arrived in Australia approximately 5 million yr ago. It is concluded here that high amounts of plasticity, in some cases adaptive, and weak integration among traits may be key to the spread and success of this species.