Cuticle micromorphology of Saxegothaea (Podocarpaceae)

The cuticle micromorphology of the leaves of the monospecific genus Saxegothaea (Podocarpaceae) was studied by scanning electron microscopy. The external and internal features of the adaxial and abaxial surfaces were characterized. The leaves are hypostomatic. The external adaxial cuticle is rugose with irregular ridges and shallow trench‐like structures that do not correspond to any feature of the inner cuticle surface. The external abaxial cuticle has densely crowded stomata arranged in two bands. The stomata are sunken with pronounced, interrupted Florin rings. Stomatal plugs were not observed. Internally, the adaxial epidermal cells are usually rectangular to square; the abaxial epidermal cells are mainly restricted to the midrib and margins and narrowly rectangular; any among the stomata are irregularly shaped. The stomata are nearly all in direct contact. They show unusual features, including an extra pair of cuticular flanges between the guard cell flanges and those of the lateral subsidiary cells, and ‘bridges’ of lateral subsidiary cell tissue extending polewards above the polar extensions to unite with those at their tips. Neither of these features has been reported previously in Podocarpaceae. The results are discussed in the light of recent phylogenetic studies. It is concluded that, despite its unique cuticular features, Saxegothaea should continue to be regarded as a member of Podocarpaceae. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 58–67.     

Foliar cuticle of Ruflorinia orlandoi nov. sp. (Pteridospermophyta) from the Lower Cretaceous of Patagonia

Foliar cuticle of a new species of Ruflorinia, R. orlandoi Carrizo et Del Fueyo, from the Lower Cretaceous Springhill Formation in Santa Cruz province, Argentina, is herein described with light and electronic (scanning and transmission) microscopy. The fronds of R. orlandoi nov. sp. are bipinnate and imparipinnate with a conspicuous main rachis bearing alternate to sub-opposite pinnae. Cuticular striations and hairs are present in both epidermises whereas stomata are only on the abaxial cuticle. Stomata have several papillate subsidiary cells and the guard cells are sunken. Ultrastructurally the adaxial and abaxial cuticles are composed of three and two cuticular layers, respectively. The cuticular characters here described are interpreted as a response to a possible adaptation to a wet climate as well as to the fluctuating environment produced by the gradual transition from fluvial to an estuarine and marine setting that characterizes the Springhill Formation. The discovery of this new species allows expanding the previous knowledge of the paleofloristic composition of the Springhill Formation with the addition of the first pteridospermous megafossil described for this formation. Moreover, the record of Ruflorinia orlandoi nov. sp. in the Springhill Formation highlights the importance of the Caytoniales during the Cretaceous in Patagonia.     

The pattern of cuticular blue-fluorescing phenolics deposition during the development of Prunus persica leaves

Although stomatal ontogeny is closely related to the development and maturation of the epidermal tissue, stomatal patterns in relation to cuticle construction and cuticular material deposition during leaf development have not received adequate attention. We observed the deposition of blue-fluorescing cuticular phenolics over guard and epidermal cells, as well as stomatal formation and patterning using the alkali-induced blue fluorescence of the cuticle of Prunus persica leaves. Stomata of different stages of maturity occurred together during leaf development, mainly at the tip of the lamina. The deposition of fluorescing compounds initially appeared over the guard cells of the developing stomata complexes and gradually extended to the neighbouring epidermal cells. Based on the blue fluorescence emitted by the cuticular layers, we constructed digital maps of leaves of different developmental stages, showing the pattern of stomatal formation and deposition of fluorescing compounds. A longitudinal tip-to-base gradient in the formation of stomata, as well as in the deposition of fluorescing compounds was observed in young developing leaves. The deposition of blue-fluorescing phenolic compounds seems to be coordinated with stomatal development.     

Heterotrophic metabolism and regulation of uptake hydrogenase activity in symbiotic cyanobacteria

The H₂ uptake activity of three cyanobionts isolated fromCycas revoluta, C. circinalis andazolla filiculoides was shown to be related primarily to the growth rate and independent of the main mode of carbon nutrition. Significant H₂ uptake was found in the coralloid roots ofCycas revoluta andZamia furfuracea (3 and 22 times higher than the respective C₂H₂ reduction activities). The results attained allow us to conclude that in cyanobacteria, in contrast to most nitrogen-fixing heterotrophs, uptake hydrogenase activity is not repressed by carbon substrates and that cyanobacteria in association seem to be endowed with sufficient H₂ uptake capacity to recover all of the H₂ released during the process of N₂-fixation.     

Stomatal function,density and pattern,and CO2 assimilation in Arabidopsis thaliana tmm1 and sdd1‐1 mutants

• Stomata modulate the exchange of water and CO₂ between plant and atmosphere. Although stomatal density is known to affect CO₂ diffusion into the leaf and thus photosynthetic rate, the effect of stomatal density and patterning on CO₂ assimilation is not fully understood.
• We used wild types Col‐0 and C24 and stomatal mutants sdd1‐1 and tmm1 of Arabidopsis thaliana, differing in stomatal density and pattern, to study the effects of these variations on both stomatal and mesophyll conductance and CO₂ assimilation rate. Anatomical parameters of stomata, leaf temperature and carbon isotope discrimination were also assessed.
• Our results indicate that increased stomatal density enhanced stomatal conductance in sdd1‐1 plants, with no effect on photosynthesis, due to both unchanged photosynthetic capacity and decreased mesophyll conductance. Clustering (abnormal patterning formed by clusters of two or more stomata) and a highly unequal distribution of stomata between the adaxial and abaxial leaf sides in tmm1 mutants also had no effect on photosynthesis.
• Except at very high stomatal densities, stomatal conductance and water loss were proportional to stomatal density. Stomatal formation in clusters reduced stomatal dynamics and their operational range as well as the efficiency of CO₂ transport.

     

The citrus leaf cuticle in relation to measurement of leaf water potential using thermocouple psychrometers*

Abstract. Cuticular resistance to water vapour diffusion is an important aspect of thermocouple psychrometry and may introduce significant error in the measurement of leaf water potential (Ψ). The effect of the citrus (Citrus mitis Blanco) leaf cuticle on water vapour movement was studied using the times required for vapour pressure equilibration during thermocouple psychrometric measurement of Ψ. Cuticular abrasion with various carborundum powders was used to reduce the diffusive resistance of both the adaxial and abaxial leaf surfaces, and the extent of the disruption to the leaf was investigated with light and electron microscopy. Cuticular abrasion resulted in reduced equilibration times due to decreased cuticular resistance and greater water vapour movement between the leaf and the psychrometer chamber. Equilibration times were reduced from over 5 h in the unabraded control leaves to 1 h with cuticle abrasion. This was associated with the decrease in diffusive resistance with cuticular abrasion from over 55 s cm^?1 to less than 8 s cm^?1 for both the adaxial and abaxial leaf surfaces. Scanning electron micrographs of the abraded leaf tissue revealed considerable disruption of the stomatal ledge and of the guard cells, surface smoothing and displacement of waxes into the stomatal aperture, and damage to veins. Observations with the transmission electron microscope revealed frequent disruption of epidermal cell walls, and damage to both the cytoplasmic and vacuolar membranes.     

Stomatal and photosynthetic responses ofCichorium intybus leaves to sulfur dioxide treatment at different stages of plant development

Fifty-day-oldCichorium intybus Linn, plants were exposed to 1 ppm sulfur dioxide gas, 2 h per day for 7 consecutive days. Their leaves as well as those from the control plants were sampled at pre-flowering, flowering, and post-flowering stages to study their morphological, physiological, and biochemical responses to SO₂ stress. The number, dimensions, area, and biomass of leaves were less in the treated plants. Length and width of stomatal apertures on both epidermises were greater for leaves exposed to SO₂. The Stomata were longer on the adaxial epidermis, but shorter on the abaxial epidermis, except at the pre-flowering stage. Stomatal widths varied widely. Compared with the controls, the abaxial epidermis on treated leaves showed consistently lower stomatal densities as well as stomatal indices. This was also true for the adaxial epidermis during the post-flowering stage. The photosynthetic rate and stomatal conductance were reduced in the SO₂-exposed plants, but intercellular CO₂ concentrations increased at the pre-flowering stage and, subsequently, declined. Chlorophyll a, carotenoid, and total chlorophyll contents increased at the pre-flowering stage, and then decreased. The level of chlorophyllb was reduced throughout plant development compared with the untreated controls.     

Association genetics,geography and ecophysiology link stomatal patterning in Populus trichocarpa with carbon gain and disease resistance trade‐offs

Stomata are essential for diffusive entry of gases to support photosynthesis, but may also expose internal leaf tissues to pathogens. To uncover trade‐offs in range‐wide adaptation relating to stomata, we investigated the underlying genetics of stomatal traits and linked variability in these traits with geoclimate, ecophysiology, condensed foliar tannins and pathogen susceptibility in black cottonwood (Populus trichocarpa). Upper (adaxial) and lower (abaxial) leaf stomatal traits were measured from 454 accessions collected throughout much of the species range. We calculated broad‐sense heritability (H²) of stomatal traits and, using SNP data from a 34K Populus SNP array, performed a genome‐wide association studies (GWAS) to uncover genes underlying stomatal trait variation. H² values for stomatal traits were moderate (average H² = 0.33). GWAS identified genes associated primarily with adaxial stomata, including polarity genes (PHABULOSA), stomatal development genes (BRASSINOSTEROID‐INSENSITIVE 2) and disease/wound‐response genes (GLUTAMATE‐CYSTEINE LIGASE). Stomatal traits correlated with latitude, gas exchange, condensed tannins and leaf rust (Melampsora) infection. Latitudinal trends of greater adaxial stomata numbers and guard cell pore size corresponded with higher stomatal conductance (g_s) and photosynthesis (A_max), faster shoot elongation, lower foliar tannins and greater Melampsora susceptibility. This suggests an evolutionary trade‐off related to differing selection pressures across the species range. In northern environments, more adaxial stomata and larger pore sizes reflect selection for rapid carbon gain and growth. By contrast, southern genotypes have fewer adaxial stomata, smaller pore sizes and higher levels of condensed tannins, possibly linked to greater pressure from natural leaf pathogens, which are less significant in northern ecosystems.     

THE CUTICULAR ANATOMY OF FRENELOPSIS VARIANS FROM THE LOWER CRETACEOUS OF CENTRAL TEXAS

Stem fragments identified as Frenelopsis varions Fontaine have been found in the Lower Cretaceous (Albian) of central Texas. The cuticle is extremely thick and characterized by 5–6 subsidiary cells with papillae overarching the stomatal chamber. Guard cells are deeply sunken below the epidermis. Stomatal complexes are arranged in axial rows extending from the base of an internode to its apex. The rows of stomata continue into the sheathing leaf where the rows curve towards the leaf apex. The epidermis of F. varions was apparently long persistent and underwent prolonged growth. Axial rows of stomata are frequently disrupted resulting in a random pattern of stomata. A single, highly reduced, sheathing leaf is present at each node. The margin of the leaf has numerous unicellular trichomes and extends to form a slightly triangular blade.     

Paraphyly of organelle DNAs in Cycas Sect. Asiorientale s due to ancient ancestral polymorphisms

Background  

This study addresses the apportionment of genetic diversity between Cycas revoluta and C. taitungensis, species that constitute the section Asiorientales and represent a unique, basal lineage of the Laurasian genus Cycas. Fossil evidence indicates divergence of the section from the rest of Cycas at least 30 million years ago. Geographically, C. taitungensis is limited to Taiwan whereas C. revoluta is found in the Ryukyu Archipelago and on mainland China.     

A new xeromorphic conifer from the Cretaceous of East China

Newly discovered coniferous shoots from the Cretaceous of Nanjing are attributed to the xeromorphic coniferous genus Glenrosa Watson and Fisher, which is found in China as well as in Eurasia for the first time. G. nanjingensis sp. nov. is known from shoots bearing scale-like leaves with thick epidermal cuticle and hypodermis. Stomata are confined to the abaxial surface deeply sunken in stomatal chambers which are filled with papillae. The cuticle, which has been sectioned for TEM, consists of an outer pale amorphous cuticle proper, usually with a peripheral polylamellate zone, and an inner dark cuticular layer containing reticulate fibrillae. G. nanjingensis sp. nov. bears a close resemblance to the type species G. texensis (Fontaine) Watson and Fisher from North America in gross morphology and especially in the characteristic stomatal chambers, but differs in phyllotaxis and in having the hypostomatic leaf with an obtuse apex. G. nanjingensis sp. nov. is believed to have grown near inland marshes or water bodies under hot and arid climates.     

Cordaites schatzlarensis sp. nov. and Samaropsis newberryi (Andrews) Seward from the Westphalian (Carboniferous) of the ?aclé? area (Czech Republic)

The monotonous cordaitalean leaves are usually difficult to determine as leaf shape and venation can be similar in many species. Therefore cuticular analysis is an important method for distinguishing cordaitalean leaves. Pennsylvanian Cordaites schatzlarensis nov. sp. comes from the ?aclé? locality in the Intrasudetic Basin, Czech Republic. It has been found in mudstone accompanying the upper coal seams No. 9 and 10 of the Jan Šverma Coals, Lampertice Member, ?aclé? Formation and are late Duckmantian in age. The leaves of Cordaites schatzlarensis are lanceolate, amphistomatic. Stomata of the adaxial epidermis are scarce, isolated, or in very short stomatal rows. In contrast, the density of stomata on the abaxial cuticle is high and stomata are arranged into single or double stomatal rows. The cuticles of C. schatzlarensis are comparable with the Chinese Upper Permian C. baodeensis Ge. Leaves can be narrow, comparable to French Bolsovian Dorycordaites zeilleri Ledran, or relatively wide. Accompanying big seeds more than 5 cm in diameter are attributed to Samaropsis newberryi (Andrews) Seward. The pith casts Artisia Sternberg were found in sandy channel fill deposits.     

Ineffectiveness of Abscisic Acid in Stomatal Closure of Yellow Lupin, Lupinus luteus var. Weiko III

Stomata of yellow lupin leaves are remarkably insensitive toabscisic acid (ABA). Stomatal resistance was monitored usingboth a viscous now porometer and a diffusion porometer. Resultswere confirmed with scanning electron microscopy. When exogenousABA solutions were supplied via petioles, 10^–6 M solutionshad no effect on stomatal resistance. Upper (adaxial) stomatawere not affected by 10^–5 M ABA but lower stomata showed3-fold more resistance after 2 h. Stomata of both surfaces closedafter 30 min in 10^–4 M ABA. Isolated epidermal peels of lupin leaves were floated on ABAsolutions yet upper surface peels showed no stomatal closingin 10^–4 M ABA, while lower surface stomata closed to abarely significant extent. Stomata of intact leaves were not very sensitive to darkness,showing at most a doubling in resistance after 6 h darkness.Complete stomatal closure, however, was readily produced bywilting leaves. Hence, lupin stomata are physically capableof closing. Endogenous ABA levels of water-stressed leaves increased approximately10-fold, which corresponds to concentrations below 10 µMABA. It is concluded that ABA is unlikely to play a role incontrolling short-term stomatal response of lupins.     

VARIATION OF STOMATAL DISTRIBUTION IN CAREX AQUATILIS (CYPERACEAE)

The density and distribution of stomates in Carex aquatilis Wahl. in the Pacific Northwest were examined using epidermal peels of samples of leaves from natural populations, from greenhouse-grown transplants and from seedling families grown under controlled conditions. These were compared to stomatal distributions of populations in eastern North America. Populations of Carex aquatilis Wahl. form 2 groups based on the distribution and density of stomates. Carex aquatilis var. dives (Holm) Kükenthal is epistomatic, with adaxial stomatal densities of 28.7–48.5/0.1 mm². The C. aquatilis var. aquatilis is amphistomatic, with adaxial stomatal densities of 8.1–22.2/0.1 mm² and abaxial densities of 11.3–24.5/0.1 mm² in the Pacific Northwest. Total stomatal frequencies are similar in both groups. Stomatal distribution and densities are here shown to not vary significantly within populations and appear to be genetically determined, as shown by progeny tests and growth of seedlings under uniform and experimental conditions. Stomatal distribution in Carex aquatilis appears to be adaptive, and intraspecific variation provides a system for determining the adaptive significance of differences in stomatal patterns.     

INTRAMARGINAL VEINED LAURACEAE LEAVES FROM THE ALBIAN–CENOMANIAN OF CHARENTE-MARITIME (WESTERN FRANCE)

Abstract:  Eucalyptolaurus depreii gen. et sp. nov. is proposed for angiosperm leaves newly collected from uppermost Albian – lowermost Cenomanian of Charente-Maritime (western France). They consist of simple, narrow, elongate laminas with entire margins and intramarginal veins. The epidermal cells of adaxial cuticle shows small, rounded, blunt papillae outward that protrude inward and fuse together as rolls along and parallel to the margins, while the adaxial cuticle bears brachyparacytic stomatal apparatus that exhibit sunken guard cells and hair bases consisting of a thick-walled pore surrounded by radially arranged differentiated cells. Resin bodies occur inside the mesophyll. These characters closely resemble the lauroid taxa ' Myrtophyllum ' and Pandemophyllum from the Cenomanian of the Czech Republic and Dakota (USA) respectively. The narrow angle of basilaminar secondaries and the whole suite of features in the guard cells (sunken guard cells embedded into subsidiary cells and stomatal ledges) strongly support close affinity with the Lauraceae. From the Cenomanian lauraceous reproductive organs and their related leaves already showed high disparity and diversity. In addition they displayed a broad ecological range from freshwater floodplains to brackish swamps. This combined to high diversity of reproductive organs suggest ecological radiation of Lauraceae by the Cenomanian.     

In situ analysis by microspectroscopy reveals triterpenoid compositional patterns within leaf cuticles of Prunus laurocerasus

The cuticular waxes on the leaves of Prunus laurocerasus are arranged in distinct layers differing in triterpenoid concentrations (Jetter et al., Plant Cell Environ 23:619–628, 2000). In addition to this transversal gradient, the lateral distribution of cuticular triterpenoids must be investigated to fully describe the spatial distribution of wax components on the leaf surfaces. In the present investigation, near infrared (NIR) Raman microspectroscopy, coherent anti-Stokes Raman scattering (CARS) microscopy, and third harmonic generation (THG) spectroscopy were employed to map the triterpenoid distribution in isolated cuticles from adaxial and abaxial sides of P. laurocerasus leaves. The relative concentrations of ursolic acid and oleanolic acid were calculated by treating the cuticle spectra as linear combinations of reference spectra from the major compounds found in the wax. Raman maps of the adaxial cuticle showed that the triterpenoids accumulate to relatively high concentrations over the periclinal regions of the pavement cells, while the very long chain aliphatic wax constituents are distributed fairly evenly across the entire adaxial cuticle. In the analysis of the abaxial cuticles, the triterpenoids were found to accumulate in greater amounts over the guard cells relative to the pavement cells. The very long chain aliphatic compounds accumulated in the cuticle above the anticlinal cell walls of the pavement cells, and were found at low concentrations above the periclinals and the guard cells.     

A new species of Cycas from the Malabar Coast, Western Ghats, India

A new cycad species,Cycas annaikalensis, has been discovered from the Malabar Coast of the southern Western Ghats of India. Although geographically distributed in a small population within the range ofC. circinalis, it can be distinguished from the other described IndianCycas species by its distinct habitat, habit, morphology of vegetative and reproductive parts, and anatomy of leaflets.     

A new species of Cycas (Cycadaceae) from Karnataka,India

Cycas swamyi Singh & Radha, sp. nov. (Cycadaceae) is described and illustrated from Karnataka, India. This new species has been confused with Cycas circinalis L., which is found in the Western Ghats. However, on the basis of its habit, characteristic isotomous‐type dichotomous branching, pinnae anatomy, mega‐ and microsporophyll morphology and seed anatomy, it has been segregated from C. circinalis and is described here as a new species. The distribution and conservation status are also discussed. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 430–435     

A Dispersed Cuticle of Cordaites from the Coal of Late Paleozoic,North China

A dispersed cuticle from the coal of Taiyuan Formation, Xuzhou Coalfield, Jiangsu Province, North China was described. It was considered as the cuticle of Cordaites because of its epidermal structures and other features evidenced by optical and scanning electronic microscopic study. Compared with the known cuticles of other species of Cordaites, it was clear that the specimen under discussion was a new type of cuticle of Cordaites. Stomata were few in number on the upper cuticle, and usually arranged in a lengthwise and intermittent file, seperated by nonstomatal cell rows. Stomatal apparatus haplocheillic nearly square in shape, 30--50 μm long and 35--55 μm wide in size and consisted of a pair of slightly sunken guard cells surrounded by 2 lateral and 2 polar subsidiary cells, orientation longitudinal and regular. Numerous stomatal apparatus on the lower cuticle were arranged mostly in a defenite file seperated by nonstomatic band with 1--10 (often 3--5) rectangular cell rows. Usually small papillae were situated on the outer periclinal wall. The guard cell was reniform and bean-shaped, 10--14 μm long and 3--5 μm wide in size. The lateral subsidiary cell was more or less rectangular or elliptical in shape, 40--125 μm long and 17—25 μm wide in size, and with papillae on the outer periclinal wall. The polar subsidiary cell was some what round, short-elliptical or some rhomboid in shape and usually shared by adjacent stomatal apparatus in the same file.