Leaf epidermis characters of Iranian <Emphasis Type="Italic">Pyrus</Emphasis> L. (Rosaceae) and their taxonomic implications

Pyrus L. is a taxonomically difficult genus mainly because of interspecific and intergeneric hybridization. A majority of the wild species of Pyrus occurs in Iran, making this region very important for any study of the systematics of the genus. In this investigation, seven quantitative and seven qualitative characters of leaf epidermis, particularly epidermal cells shape, anticlinal walls patterns, type and size of stomata, and cuticular ornamentation of 22 taxa of Pyrus plus Malus orientalis are studied for assessment of their taxonomical value at different levels. Scanning electron microscopy and light microscopy were used for evaluation of epidermis features of these taxa. Some characters such as shape and anticlinal wall pattern of epidermal cells, presence of adaxial stomata, pattern of cuticular ornamentation and epicuticular wax types are useful in separation of some Pyrus taxa. Moreover, Malus orientalis is recognized easily from all Pyrus species via the density of stomata. According to ANOVA analysis, all quantitative variables show high level of between-group differentiation. Also, based on the hierarchical clustering results gained by multiple correspondence analysis and factor analysis of mixed data analyses of the current study, micromorphological characters of epidermal cells reflect the ecological conditions of pear taxa, so most of the mesophyte and xerophyte taxa can be separated from each other in accordance with epidermal cells characters, specially the anticlinal wall pattern. Totally, the application of some other biological markers, particularly suitable molecular data are needed to present well-resolved relationships among these taxa.     

Ultraviolet leaf reflectance of common urban trees and the prediction of reflectance from leaf surface characteristics

The spectral reflectance and transmittance over the wavelength range of 250–700 nm were evaluated for leaves of 20 deciduous tree species and leaf sheaths of five isogenic wax variants of Sorghum bicolor differing in visible reflectance due to cuticular waxes. Using the sorghum sheath reflectance and cuticle surface characteristics as a model, it was concluded that tree leaf reflectance above 0.06 was likely due to the presence of variously-shaped fine epicuticular wax structures on the leaf surface. Increasing the density of sub-micron wax structures corresponded to an enhanced ultraviolet (UV) reflectance over the PAR reflectance of a given leaf surface—either S. bicolor sheath or tree leaf. Amorphous globular epicuticular wax structures did not appear to scatter UV as well as wax filaments or vertical plates in varying patterns even when the dimensions of the structures were similar. Further work is needed to clarify this relationship and the influence of cellular pigments on subsurface contributions to the reflectance.     

Distribution of folpet on the grape surface after treatment

Field trials were carried out to evaluate whether folpet sprayed on grapevines penetrated the epicuticular wax and cell walls of grapes. Folpet showed poor penetration into the epicuticular wax; it was found almost totally on the surface. Despite its low solubility in water, perhaps due to the presence of adjuvants, its residues showed such a high resistance to washing that the action of rain was negligible in decreasing residues.     

Genetic diversity assessment for Eugenia uniflora L., E. pyriformis Cambess., E. brasiliensis Lam. and E. francavilleana O. Berg neotropical tree species (Myrtaceae) with heterologous SSR markers

In this study, a set of nuclear microsatellite markers were transferred and characterized from two Myrtaceae sub-families to four neotropical Eugenia fruit tree species (E. uniflora L., E. pyriformis Cambess., E. brasiliensis Lam. and E. francavilleana O. Berg), which are neglected and underutilized foods of great ecological and potential economic value found in the Atlantic forest regions of South America. Leaf samples of mature E. uniflora, E. pyriformis, E. brasiliensis and E. francavilleana trees, which are popularly termed “pitanga”, “uvaia”, “grumixama” and “guamirim”, respectively, were collected from two areas greatly impacted by the agricultural practices of the sugar cane industry in the state of São Paulo in the southeastern region of Brazil. A total of 19 simple sequence repeat (SSR) markers from the Myrtaceae sub-families Leptospermoideae and Myrtoideae were tested, and 15 polymorphic heterologous SSR markers were identified. The expected heterozygosities for E. uniflora, E. pyriformis, E. brasiliensis and E. francavilleana were 0.64, 0.75, 0.54 and 0.71, respectively. These results suggest that these SSR markers may be useful for population genetic studies. A total of 4, 9, 11 and 4 heterologous SSR polymorphic markers for E. uniflora, E. pyriformis, E. brasiliensis and E. francavilleana, respectively, are now available for genetic diversity, gene flow and mating system analyses in these species.     

Epicuticular Wax of Norway Spruce Needles as Indicator of Traffic Pollutant Deposition

Two-year-old Norway spruce (Picea abies (L.) Karst) seedlings were exposed to trafficemissions along roadsides with three different trafficdensities and speed limits; highway, street and aquiet local road. Deposition of the polyaromatichydrocarbons (PAH) was determined at all sites usingmoss bags as an indicator of organic traffic pollutantdeposition. Short term concentrations of atmosphericnitrogen oxides (NO_x) and aerosol black carbon(BC) were also recorded from the sites. The responsesof the needle epicuticular wax structure of theexposed spruce seedlings were analysed under thescanning electron microscopy (SEM). The epistomatalwax structure of needles was distinctively fused andaggregated along the highway and street withrelatively high traffic densities. Along the localroad, where the traffic density was low, nosignificant degradation of needle epicuticular wax wasobserved. The NO_x and BC data, along with PAHsaccumulated in moss bags indicate significantdeposition of gaseous pollutants and particulateorganic compounds along the highway and street andthese are suggested as cause of the epicuticular waxdegradation. Epistomatal wax structure is thus suggested to be an early indicator of traffic exposure effects.     

Annual Changes in Some Physiological Properties of Cryptomeria Japonica Leaves from Kanto, Japan

C. japonica leaves were sampled monthly in a heavily damaged area (a-I: Saitama), a slightly damaged area (a-II: plains in Ibaraki), and a healthy area (a-III: mountainous areas in Ibaraki) in Kanto. The leaves were analyzed for apparent cuticular transpiration rates, amounts of epicuticular wax, and contact angles. Sb in aerosols deposited on the leaves was also analyzed. The transpiration rates and the increase in transpiration with leaf aging were higher in a-I than in a-II and a-III. Erosion rates of the wax were higher in a-I and a-II than in a-III. The decrease of contact angles with aging tended to be higher in a-I and a-II than in a-III. Rates of aerosol-Sb deposition on the leaves were in the order a-I ? a-II > a-III. The transpiration rates correlated with the values obtained from a linear binomial function that included the amounts of wax and aerosol-Sb as variables (r = 0.855, P < 0.01). In a-I, the large quantity of aerosols on the leaves (and probably gaseous air pollutants) may have increased the transpiration rates by ca. 50% owing to erosion of the wax and stomatal malfunction, placing C. japonica under chronic water stress.     

干旱胁迫对水稻叶表面蜡质积累的影响

环境胁迫一直以来是影响植物生长和农作物产量的主要因素，而覆盖植物叶片表层的蜡质层在植物生长发育和适应外界环境方面起着关键作用。本试验以中花11、广盐2号和Z46这3个水稻品种进行干旱胁迫处理，通过扫描电镜观察叶片表面蜡质覆盖密度，以及测定叶片相对含水量、叶绿素浸提率以及失水率等生理指标，从而综合分析干旱胁迫对不同水稻品种叶片表面蜡质积累的影响。结果表明：干旱胁迫条件下，3个试验的水稻品种叶片相对含水量降低，叶表面蜡质覆盖量增加，叶绿素浸提率以及离体叶片失水率下降。     

Salinity-induced changes in biometric,physiological and anatomical parameters of Passiflora edulis Sims plants propagated by different methods

ABSTRACT

Salinity is one of the most important abiotic stresses limiting agricultural production worldwide. Here, we investigated the biometric, physiological and anatomical changes of Passiflora edulis plants propagated from seeds, cuttings and grafts and exposed to salt stress (0.3 – control and 6.3 dS m^?1). Saline stress significantly reduced plant height, the number of leaves, total chlorophyll, stomatal conductance, shoot dry weight, total dry weight, total water consumption and root volume. Among the propagation methods evaluated, plants propagated from seeds were more sensitive to salinity, while plants propagated from cuttings showed smaller alterations when subjected to salinity. Similar behavior was observed for anatomical characteristics, since plants propagated from seeds presented more pronounced anatomical alterations, such as greater thicknesses of the leaf mesophyll, upper epidermis, lower epidermis, palisade parenchyma and spongy parenchyma and also was observed presence of calcium oxalate crystals in the form of druses in greater quantity in the root cortex, in comparison to plants propagated from cuttings. The approach used in this study allowed identifying the type of yellow passion fruit propagation (cuttings) that was best for physiological and anatomical adaptation to salt stress and will be potentially useful in programs for improvement of the crop.     

Leaf epicuticular wax as a factor of antixenotic resistance of cabbage to cabbage flea beetles and cabbage stink bugs attack

The aim of present research was to establish the role of epicuticular wax content in eight cabbage genotypes (four white hybrids and one red hybrid, two red varieties and one white variety) in the context of its natural resistance to attack cabbage flea beetles (Phyllotreta spp.) and cabbage stink bugs (Eurydema spp.), which are among the most important cabbage pests in southern Europe. For this reason and for the purpose of diminishing the use of synthetic insecticides against the cabbage pests the field experiments in 2006 and 2008 were conducted. We found out that individual cabbage genotypes – they had different epicuticular wax content – differ in regard to their susceptibility to attacks by the studied groups of harmful insect pests. The highest susceptibility to attacks by Phyllotreta spp. was confirmed for the hybrid ‘Cheers F1’, in the first year (1.68 ± 0.05), as well as in the second year of the experiment (2.87 ± 0.13). Cabbage stink bugs in both years of the experiment caused the highest extent of injuries on the hybrids ‘Destiny F1’, ‘Cheers F1’, and ‘Vestri F1’. In both years we found higher epicuticular wax content in red cabbage genotypes. In almost all studied genotypes we found a pronounced negative correlation between the content of epicuticular wax and the extent of injuries done by both groups of harmful pests. We have established that epicuticular wax is an important factor of cabbage's antixenotic resistance to attacks by cabbage flea beetles and cabbage stink bugs, and that the cabbage genotypes with higher content of this substance are consequently more suitable for environmentally acceptable manners of cabbage production.     

水稻叶片上下表面反射率差异及其与氮素状况的关系

在田间条件下测定了5个氮素水平处理的水稻叶片在分蘖、孕穗和抽穗期的上表面反射率和下表面反射率。结果表明:各生育期水稻叶片在可见光和近红外区域的下表面反射率均大于上表面反射率(绝对差值一般小于2%),上下表面反射率差值大小与供氮水平间无显著相关性,但在近红外区域这种差值随着生育期而降低。与上表面反射率相比,下表面反射率与叶片氮素和叶绿素含量间具有同样高的相关性,因此水稻下表面反射率也可用于估计叶片氮素和叶绿素含量。下表面中脉向外凸出及上下表皮细胞大小与排列的差异可能是引起水稻叶片上下表面反射差异的主要原因     

Microdistribution of aluminum and manganese in the tea leaf tissues as revealed by X‐ray microanalyzer

Abstract

Tea plants (Thea sinensis L.) were found to accumulate large amount of Al (4457 ppm, D.W.) in their old leaves. The results showed that Mn stimulated the Al uptake and increased the Al content in the leaves. High concentrations of Ca and Mg in the soils decreased the Al and Mn and increased the Ca and Mg contents in the leaves. No phosphorus and aluminum interaction was found in the leaves of tea plants.

Microdistribution patterns of Al and Mn in the tea leaf tissues were also studied by means of a wavelength dispersive type X‐ray microanalyzer in conjunction with cryostage. The results of X‐ray micrographs showed that Al was densely deposited on the cell walls of the adaxial epidermis and palisade parenchyma cells of the old leaf tissues, and this distribution pattern of Al was different than that of Mn which was found mostly concentrated in the epidermal cells.     

Silicon Decreases Transpiration Rate and Conductance from Stomata of Maize Plants

ABSTRACT

To characterize the effect of silicon (Si) on decreasing transpiration rate in maize (Zea mays L.) plants, the transpiration rate and conductance from both leaves and cuticula of maize plants were measured directly. Plants were grown in nutrient solutions with and without Si under both normal water conditions and drought stress [20% polyethylene glycol (PEG) concentration in nutrient solution] treatments. Silicon application of 2 mmol L^?1 significantly decreased transpiration rate and conductance for both adaxial and abaxial leaf surface, but had no effect on transpiration rate and conductance from the cuticle. These results indicate that the role of Si in decreasing transpiration rate must be largely attributed to the reduction in transpiration rate from stomata rather than cuticula. Stomatal structure, element deposition, and stomatal density on both adaxial and abaxial leaf surfaces were observed with scanning electron microscopy (SEM) and a light microscope. Results showed that changes in neither stomatal morphology nor stomatal density could explain the role of Si in decreasing stomatal transpiration of maize plants. Silicon application with H₄SiO₄ significantly increased Si concentration in shoots and roots of maize plants. Silicon concentration in shoots of maize plants was higher than in roots, whether or not Si was applied. Silicon deposits in cell walls of the leaf epidermis were mostly in the form of polymerized SiO₂.     

Phosphorus status of paddy soils from the central region of the Mekong River in Laos

Microorganisms colonizing the leaf sheaths of rice plants placed in a waterlogged paddy field were studied using a scanning electron microscope. Decomposition was faster on the adaxial side than on the abaxial side of the leaf sheaths. The veins and trichomes were highly resistant to microbial decomposition throughout the waterlogged period of rice growth. The decomposition and disintegration of leaf sheaths markedly progressed from the third month (from July) after the placement in the paddy field. Microbial colonization was gradual in the first month after the placement of leaf sheaths in the field. One of the common first colonizers had a terminal endospore (Clostridium dissolvens-like microorganisms). Microbial colonization proceeded actively in the second month, and microbiota became diversified. Microbial colonization on the adaxial surface was more than that on the abaxial surface. Respective microorganisms colonized the surfaces separately from each others in this period. They grew mainly on the thin membrane that covered the epidermal layer. Microbial colonies spread all over the abaxial and adaxial surfaces, and there was no significant difference in the degree of colonization between the abaxial surface and the adaxial surface in the third month (in July). Although separate colonization of the respective microorganisms was common, the co-existence of different microorganisms within a colony was also observed. The thin membrane that covered the epidermal layer and the epidermis seemed to be the main substrate during this period. Co-existence of different microorganisms was a common occurrence in colony formation, and organic debris and remaining tissues of leaf sheaths were assumed to be the major substrates in August. These findings on microbial colonization were well correlated with the findings on the degree of decomposition.     

Differences in Iron, Manganese, and Phosphorus Binding in Freshwater Sediment Vegetated with Littorella Uniflora and Benthic Microalgae

Undisturbed sediment cores from an oligotrophic lake were percolated with artificial porewater to examine the effects of isoetid macrophytes,Littorella uniflora, and benthic microalgae on daily dynamics of sediment retention of phosphorus (P) by either iron (Fe) or manganese (Mn). Retention of Fe and Mn was observed due to oxidation processes mediated by oxygen release fromL. uniflora roots and benthic microalgae. Therefore increased retention of P was observed because of P precipitation with oxidized Fe- and Mn-compounds. During light periods, the ratio between Fe and P precipitation in the sediment was positively correlated with the P uptake byL. uniflora (p < 0.001, r² = 0.984). The atomic precipitation ratio between Fe and P was between I and 2. The ratio between oxidized Fe-compounds and Fe-bound phosphate in the sediment was positively correlated with the root density ofL. uniflora (p < 0 001. r² = 0.995). The ratio between Mn and P precipitation was higher (26) than the ratio between Fe and P. The role of benthic primary producers on P retention in freshwater littoral sediments is discussed.     

Permeability of peach leaf cuticles to boron

Isolated leaf cuticles and intact plants of one‐year‐old potted ‘Red Haven’ trees grown in the greenhouse were used to study the permeability of leaf cuticles to boron (B). The isolated leaf cuticles were prepared enzymatically. Flux was determined using a transport cell. Permeability coefficient (P) was calculated and used as the parameter of cuticle permeability to B. For intact plant studies, enriched‐¹⁰B boric acid solution was evenly spread onto the abaxial or the adaxial side of a leaf. The trees were sacrificed three days after the treatment and selected plant parts were analyzed with ICP‐AES and ICP‐MS spectrometers. The ¹⁰B content was used as the parameter of B permeability in vivo. The permeability coefficient of peach leaf adaxial cuticle was in the order of 10^‐7 cm/sec. There were no difference in the permeability coefficients among B concentration and surtactant treatments. The pH had variable effect on B absorption through the cuticle of the adaxial side of the leaf. For the in vivo study of B uptake by peach leaves, the abaxial side was more efficient than the adaxial side, with the total ¹⁰B taken up four times greater in the abaxial side than the adaxial side. The absorption percentages of the total ¹⁰B applied in the abaxial and adaxial sides were 0.31% and 0.08%, respectively. Scratches on the treated leaves made the total ¹⁰B uptake and translocation higher than the control treatment.     

Effects of Tropospheric O3 on Trembling Aspen and Interaction with Co2: Results from an O3-Gradient and a Face Experiment

Over the years, a series of trembling aspen (Populus tremuloides Michx.) clones differing in O₃ sensitivity have been identified from OTC studies. Three clones (216 and 271[(O₃ tolerant] and 259 [O₃ sensitive]) have been characterized for O₃ sensitivity by growth and biomass responses, foliar symptoms, gas exchange, chlorophyll content, epicuticular wax characteristics, and antioxidant production. In this study we compared the responses of these same clones exposed to O₃ under field conditions along a natural O₃ gradient and in a Free-Air CO₂ and O₃ Enrichment (FACE) facility. In addition, we examined how elevated CO₂ affected O₃ symptom development. Visible O₃ symptoms were consistently seen (5 out of 6 years) at two of the three sites along the O₃ gradient and where daily one-hour maximum concentrations were in the range of 96 to 125 ppb. Clonal differences in O₃ sensitivity were consistent with our OTC rankings. Elevated CO₂ (200 ppm over ambient and applied during daylight hours during the growing season) reduced visible foliar symptoms for all three clones from 31 to 96% as determined by symptom development in elevated O₃ versus elevated O₃ + CO₂ treatments. Degradation of the epicuticular wax surface of all three clones was found at the two elevated O₃ gradient sites. This degradation was quantified by a coefficient of occlusion which was a measure of stomatal occlusion by epicuticular waxes. Statistically significant increases in stomatal occlusion compared to controls were found for all three clones and for all treatments including elevated CO₂, elevated O₃, and elevated CO₂ + O₃. Our results provide additional evidence that current ambient O₃ levels in the Great Lakes region are causing adverse effects on trembling aspen. Whether or not elevated CO₂ in the future will alleviate some of these adverse effects, as occurred with visible symptoms but not with epicuticular wax degradation, is unknown.     

Effects of simulated rain acidified with sulfuric acid on host-parasite interactions

Wind-blown rain, rain splash, and films of free moisture play important roles in the epidemiology of many plant diseases. The effects of simulated rain acidified with sulfuric acid were studied on several host-parasite systems. Plants were exposed in greenhouse or field to simulated rain of pH 3.2 ± 0.1 or pH 6.0 ± 0.2. Simulated ‘rain’ of pH 3.2 resulted in: (1) an 86% inhibition in telia production of Cronartium fusiforme on Quercus phellos; (2) a 66% inhibition of reproduction of Meloidogyne hapla on field-grown Phaseolus vulgaris; (3) a 10% decrease in the severity of Uromyces phaseoli on field-grown Phaseolus vulgaris; and (4) an inhibition of Rhizobium nodulation of Phaseolus vulgaris and Glycine max by an average of 73%. Effects on halo blight of kidney bean (caused by Pseudomonas phaseolicola) depended upon the segment of the disease cycle in which the ‘rain’ occurred: (a) simulated rain of pH 3.2 applied to plants before inoculation stimulated disease development; (b) suspension of inoculum in ‘rain’ of pH 3.2 decreased inoculum potential; and (c) ‘rain’ of pH 3.2 applied to plants after infection inhibited disease development. Scanning electron microscopy of epicuticular waxes on leaves of Quercus phellos and Phaseolus vulgaris showed marked erosion of those surfaces by ‘rain’ of pH 3.2, indicating possible influences on the structure and function of plant cuticles. These results suggest that the acidity of rain is a new parameter of environmental concern, and underline the need for study of the consequences of prolonged exposure of both agronomic and natural ecosystems to this stress factor.     

Endophytic nitrogen-fixing Klebsiella variicola strain DX120E promotes sugarcane growth

Klebsiella variicola bacteria are found in association with plants. Little is known about their colonization patterns, roles, and mechanisms during association with the plant hosts. Here, we identified a nitrogen-fixing bacterium, DX120E, which was isolated from surface-sterilized roots of the ROC22 sugarcane cultivar, as K. variicola by phylogenetic analyses of its 16S rRNA gene, RNA polymerase β-subunit gene, and DNA gyrase subunit A gene sequences. gfp-tagged DX120E was found to colonize at the roots and aerial parts of micropropagated sugarcane plantlets by fluorescence microscopy and confocal microscopy. DX120E was able to survive in soils and colonize in root epidermal cells, intercellular spaces in root cortices, and leaf mesophyll and vascular tissues. DX120E preferentially colonized at root maturation and elongation zones and entered roots via cracks at the emergence site of lateral roots and at disrupted epidermis. DX120E may penetrate root epidermal cells with the aid of their cellulose-degrading enzymes. ¹⁵N isotope dilution assay demonstrated that DX120E was able to fix N₂ in association with ROC22 sugarcane plants under gnotobiotic condition. DX120E was also able to promote GT21 cultivar growth and plant uptake of N, P, and K under greenhouse condition. Together, this study for the first time shows that a K. variicola strain is able to colonize in its sugarcane plant hosts, to fix N₂ in association with plants, and to promote plant growth.     

Structural Analysis of Tillandsia usneoides L. Exposed to Air Pollutants in São Paulo City–Brazil

Bioindicators plants are important for the evaluation of air quality and Tillandsia usneoides L., an atmospheric epiphyte bromeliad, has been used for this purpose. The present study aims at evaluate the structural pattern of the leaf of this species when exposed to urban air pollutants, and determining whether the leaves present structural parameters that could be used as indicators of such pollutants. Samples of T. usneoides were exposed in São Paulo, the biggest city of Brazil, for 8, 16 and 24 weeks, and compared with others kept in a rural area. The urban pollution of São Paulo affected the structure of the leaves of T. usneoides causing alterations, especially in the scales, density of stomata and epidermis thickness. Qualitative alterations in the mesophyll were not observed in plants exposed at the polluted sites. These structural characteristics of T. usneoides seem to account for its high tolerance to heavy metal accumulation. The percentage of anomalous scales may potentially be used as an alternative bioindicator parameter.     

Needle Structures and Epiphytic Microflora of Scots Pine (Pinus Sylvestris L.) under Heavy Ammonia Deposition from Fur Farming

Scots pine (Pinus sylvestris) needles from forest sites differing in distance from big fur farms emitting large amounts of ammonia and ammonium (=NH_y) were examined by scanning and transmission electron microscopy. Specific features indicating an ammonium-nitrogen overload, such as an abundance of needle surface organisms and modifications in cellular and wax structures, were classified. Throughout the study area (up to 1000 m from the farms), mesophyll cells had thin cytoplasm and folding plasmalemma indicating frost damage. Phloem damage attributable to a possible nutrient imbalance was also observed. Chloroplast membranes were undulating and the occurrence of leaf surface organisms (e.g. aerophilic algae) was more abundant at the closest sites. The changes were related both to the direct effects of dry NH_y deposition on the needles, and to the effects operating via soil acidification. The needle epicuticular waxes proved to be structurally rather inert against the influence of ammonium compounds, since no significant changes due to NH_y were observed.