Studies on Leaf Surface Lipid of Tobacco I. Changes in Leaf Surface Lipid and Duvatrienediol during Growth,Senescence and Curing of Tobacco Leaves

Duvatrienediol is a diterpene specifically occurred in tobacco plants and thought to be a precursor of tobacco aroma. Green tobacco leaves contained 0.2~1% of duvatrienediol per dry weight and it was corresponded to 30~60% of leaf surface lipid. Leaves on upper stalk position contained more of leaf surface lipid and duvatrienediol. In leaves on each stalk position, leaf surface lipid and duvatrienediol contents increased with leaf growth and decreased by over-maturation. Production of leaf surface lipid and duvatrienediol was affected by soil conditions or applied amount of nitrogen fertilizer. Both leaf surface lipid and duvatrienediol were decreased during curing of tobacco leaves, but the change in the latter was more drastic. Comparing to leaf surface lipid, changes in cytoplasmic lipid were less during growth and senescence of tobacco leaves.     

Changes of epicuticular wax induced by enhanced UV-B radiation impact on gas exchange in Brassica napus

The epicuticular wax covering on plant surface plays important roles in protecting plants against UV radiation. However, the role of epicuticular wax in affecting leaf gas exchange under enhanced ultraviolet-B (UV-B) radiation remains obscure. In the present study, different aged leaves of Brassica napus were used to analyze the responses of crystal structure and chemical constituents of epicuticular wax to UV-B radiation and the effects of such responses on gas exchange indices. Enhanced UV-B radiation significantly decreased the amount of esters in all leaves except the first leaf, amount of secondary alcohols in the second, third and fourth leaves, and amount of primary alcohols in the second and third leaves, while increased the amounts of ketones and aldehydes in the first leaf. Enhanced UV-B level had no significant effect on the amounts of alkanes and total wax in all leaves. Exposure to UV-B radiation resulted in wax fusion on adaxial leaf and stomata opening on abaxial leaf. Fusions of plates and rods on adaxial leaf surface covered most of the stomata, thereby influencing the photosynthesis in the upper mesophyll of leaves. Enhanced UV-B level significantly reduced the net photosynthesis rate (P _N) but increased the stomata conductance (g _s), concentrations of intercellular CO₂ (C _i ), and transpiration rate (E) in all leaves. Both UV-B radiation and the wax fusion induced by enhanced UV-B radiation resulted in different stomata status on abaxial and adaxial leaf surface, causing decrease of P _N, and increase of g _s, C _i and E in leaves.     

The impact of epicuticular wax on gas-exchange and photoinhibition in Leucadendron lanigerum (Proteaceae)

This study investigated the seasonal modification of wax deposition, and the impact of epicuticular wax on gas-exchange as well as photoinhibition in Leucadendron lanigerum, a species from the Proteaceae family with wax-covered leaf surfaces and the stomata also partially occluded by wax. The results of this study demonstrated that the deposition of epicuticular wax in L. lanigerum is dependent on the age of the leaf as well as the season, and generation and regeneration of wax occur mostly in spring while transformation and also degeneration of wax crystals occur in winter. Epicuticular waxes decreased cuticular water loss, but had little impact on leaf reflectance. The temperature of leaves without wax was lower than that of wax-covered leaves, indicating that the rate of transpiration impacted more on leaf temperature than reflectance of light in the PAR range in L. lanigerum. The wax coverage at the entrance of stomata in L. lanigerum increased resistance to gas diffusion and as a consequence decreased stomatal conductance, transpiration and photosynthesis. Also, the results indicated that epicuticular waxes do help prevent photodamage in L. lanigerum, and so this property could benefit plants living in arid environments with high solar radiation.     

Changes in leaf epicuticular wax,gas exchange and biochemistry metabolism between <Emphasis Type="Italic">Jatropha mollissima</Emphasis> and <Emphasis Type="Italic">Jatropha curcas</Emphasis> under semi-arid conditions

Jatropha curcas and Jatropha mollissima plants were evaluated under conditions of high (HSM) and low (LSM) soil moisture in a semi-arid environment, as changes in the content and concentration of epicuticular wax and the leaf metabolism which could have a relationship with drought tolerance. Besides epicuticular wax, gas exchange, antioxidant system and biochemical parameters of the photosynthetic metabolism were measured. The epicuticular wax content increased only in J. mollissima leaves 95 % under LSM, when compared with HSM conditions. Therefore, J. curcas invested less in the production of long-chain n-alkanes than did J. mollissima under LSM conditions. J. mollissima plants showed the highest CO₂ assimilation rate during the HSM period compared to J. curcas. Both species showed high stability in some leaf biochemistry products, highlighting the highest sugar content, free amino acids, total soluble protein, and photosynthetic pigments in the leaves of J. mollissima plants under both of the soil moisture conditions. Moreover, the stability and performance of the different parameters, such as morphologic variables, seem to allow J. mollissima plants to tolerate semi-arid conditions.     

Sclerophylly in Qualea Parviflora (Vochysiaceae): Influence of Herbivory, Mineral Nutrients, and Water Status

Qualea parviflora Mart. (Vochysiaceae) is a deciduous tree, commonly observed in campo sujo, cerrado sensu stricto and cerradão vegetation types in Brazilian cerrado (savannas). In this study we investigated herbivory, nutritional, and water status effects on leaf sclerophylly of Q. parviflora. Twenty fully expanded leaves were taken from 10 plants in each vegetation type four times a year. Mean leaf concentration of N, P, K, Ca, C, Al, Si, and percentage of total phenols, herbivory and tannins were measured on a plant basis. Leaf specific mass (LSM) (g m^?2), a sclerophylly index, and pre-dawn leaf water potential (MPa) were also recorded. Soil samples below each tree were collected to quantify N–NO₃, N–NH₄, P, K, Mn, soil moisture, organic matter, Si, and Al. Qualea parviflora showed a LSM from 69 to 202 g m^?2 and leaves were younger and less sclerophyllous in November (beginning of rainy season). Q. parviflora inhabiting the cerradão had leaves with higher concentration of nutrients and lower sclerophylly while trees in campo sujo and cerrado sensu stricto did not show significant differences in leaf sclerophylly. The concentrations of N, P, K and tannins had an inverse relationship with leaf age. Concentration of phenols, Al, C, Ca, Si, C/N and Ca/K increased with leaf age. The concentrations of P and Ca/K ratio in leaves explained 60% of variation observed in leaf sclerophylly. We did not find any significant relationship between the level of sclerophylly and water potential or herbivory. Our results corroborate the hypothesis that predicts lower concentrations of essential macronutrients would be the main factors influencing higher sclerophylly in leaves of Q. parviflora plants in Cerrado.     

Duvatrienediols in cuticular wax of Burley tobacco leaves.

4, 8, 13-Duvatriene-1, 3-diol diastereoisomers have been identified in the cuticular wax of fresh Burley tobacco leaves. Their structures were determined by gas-liquid chromatography, mass spectrometry, and infrared and ultraviolet spectroscopy. Butylboronic acid derivatives of the alpha, beta-isomers were separated by gas-liquid chromatography and identified by mass spectrometry. The quantitative determination by gas-liquid chromatography revealed that the duvatrienediols are major components in the cuticular wax of Nicotiana tabacum. The duvatrienediol content in young leaves is higher than in old leaves, and in young leaves this compound may account for half of the cuticular wax.     

Variations of cuticular wax in mulberry trees and their effects on gas exchange and post-harvest water loss

Though mulberry (Morus alba) tree shows great adaptations to various climate conditions, their leaf water status and photosynthesis are sensitive to climate changes. In the current study, seven widely planted mulberry cultivars in Chongqing, Southwest China, were selected to analyze leaf cuticular wax characteristics, gas exchange index, post-harvest leaf water status and their relationships, aiming to provide new theory in screening high resistant mulberry cultivars. Mulberry trees formed rounded cap-type idioblasts on the adaxial leaf surface. Film-like waxes and granule-type wax crystals covered leaf surfaces, varying in crystal density among cultivars. The stomatal aperture on the abaxial surface of cultivars with high wax amount was smaller than that of cultivars with low wax amount. The amount of total wax was negatively correlated with the net photosynthetic rate (P _N), transpiration rate (E) and stomatal conductance (g _s) and positively correlated with the moisture retention capacity. It suggested that both cuticular wax and stomatal factor might be involved in regulating water loss in mulberry leaves under field conditions. The variability in moisture retention capacity and cuticular wax characteristics might be important in evaluating and screening mulberry cultivars for increasing silk quality and silkworm productivity.     

Changes in leaf morphology and anatomy with tree age and height in the broadleaved evergreen species, Eucalyptus regnans F. Muell

Relatively little is known about changes in leaf attributes over the lifespan of woody plants. Knowledge of such changes may be useful in interpreting physiological changes with age. This study investigated changes in leaf morphology and anatomy with tree age and height in the broadleaved evergreen species, Eucalyptus regnans. Fully expanded leaves were sampled from the upper canopy of tree ages ranging from 6 to 240 years, and tree heights ranging from about 10–80 m. There were significant changes in leaf form with increasing tree age and height. Leaf size and specific leaf area (SLA; leaf area/leaf mass) decreased, leaf thickness increased, and leaves became narrower relative to their length, with increasing tree age and height. Cuticle thickness and leaf waxiness, including wax occlusion of the stomatal antechamber, increased with increasing age and height. By comparison, there were no clear trends in stomatal frequency or stomatal length with tree age, although there were curvilinear relationships between an index of total stomatal pore area per leaf lamina and both tree age and tree height. The results support the hypothesis that leaves of E. regnans become more xeromorphic with tree age and height. The results are discussed in relation to their significance for changes in water relations in the canopy with age.     

BraLTP1, a Lipid Transfer Protein Gene Involved in Epicuticular Wax Deposition,Cell Proliferation and Flower Development in Brassica napus

Plant non-specific lipid transfer proteins (nsLTPs) constitute large multigene families that possess complex physiological functions, many of which remain unclear. This study isolated and characterized the function of a lipid transfer protein gene, BraLTP1 from Brassica rapa, in the important oilseed crops Brassica napus. BraLTP1 encodes a predicted secretory protein, in the little known VI Class of nsLTP families. Overexpression of BnaLTP1 in B. napus caused abnormal green coloration and reduced wax deposition on leaves and detailed wax analysis revealed 17–80% reduction in various major wax components, which resulted in significant water-loss relative to wild type. BnaLTP1 overexpressing leaves exhibited morphological disfiguration and abaxially curled leaf edges, and leaf cross-sections revealed cell overproliferation that was correlated to increased cytokinin levels (tZ, tZR, iP, and iPR) in leaves and high expression of the cytokinin biosynthsis gene IPT3. BnaLTP1-overexpressing plants also displayed morphological disfiguration of flowers, with early-onset and elongated carpel development and outwardly curled stamen. This was consistent with altered expression of a a number of ABC model genes related to flower development. Together, these results suggest that BraLTP1 is a new nsLTP gene involved in wax production or deposition, with additional direct or indirect effects on cell division and flower development.     

广州市常见行道树种叶片表面形态与滞尘能力

以广州市常见的18种行道树为对象,通过扫描电镜观察比较了行道树的叶表面形态结构、应用接触角测定仪测定了绿化树种叶片的接触角对滞尘能力的影响.结果表明:不同树种的滞尘量差异显著,18种植物叶片雨后第26天的最大滞尘量在0.066-1.831 g/m2,物种间相差达27倍以上.叶表面具有网状结构,气孔密度较大(20＜气孔密度＜60个)且气孔开口较大(如芒果)容易滞留粉尘;叶表面平滑具有蜡质层,气孔排列整齐,无明显起伏(如红花羊蹄甲、桃花心木、大叶紫薇、鹅掌藤),滞尘能力较弱.植物叶片接触角与滞尘量呈负相关(r=-0.614),接触角＜90°的表现为亲水性.易润湿的植物叶片雨后第26天最大滞尘量在1.0-1.831 g/m2,叶片表面的形态结构凹凸不平,具有钩状或脊状褶皱、突起等且20＜气孔密度＜60范围内,测得的接触角较小(芒果、重阳木、高山榕),使得粉尘与植物叶片接触面积较大,粉尘不易从叶面脱落,滞尘能力较强.而接触角较大的盆架树、麻楝、大叶紫薇、鹅掌藤和红花羊蹄甲的滞尘量均＜1.0g/m2,其特殊的表面结构和疏水的蜡质使颗粒物不易吸附在植物叶片上,因此滞尘能力较弱.由此可见,植物叶表面蜡质含量和气孔密度及其叶片接触角的大小是影响植物叶片滞尘能力的主要因素,在进行城市绿化时,适当考虑选择叶表面形态有利于滞尘的绿化树种,将可提高城市植被的环境效应.     

Abscisic Acid Content, Transpiration, and Stomatal Conductance As Related to Leaf Age in Plants of Xanthium strumarium L

Among the four uppermost leaves of greenhouse-grown plants of Xanthium strumarium L. the content of abscisic acid per unit fresh or dry weight was highest in the youngest leaf and decreased gradually with increasing age of the leaves. Expressed per leaf, the second youngest leaf was richest in ABA; the amount of ABA per leaf declined only slightly as the leaves expanded. Transpiration and stomatal conductance were negatively correlated with the ABA concentration in the leaves; the youngest leaf lost the least amount of water. This correlation was always very good if the youngest leaf was compared with the older leaves but not always good among the older leaves. Since stomatal sensitivity to exogenous (±)-ABA was the same in leaves of all four age groups ABA may be in at least two compartments in the leaf, one of which is isolated from the guard cells.     

Assessing the Capacity of Plant Species to Accumulate Particulate Matter in Beijing,China

Air pollution causes serious problems in spring in northern China; therefore, studying the ability of different plants to accumulate particulate matter (PM) at the beginning of the growing season may benefit urban planners in their attempts to control air pollution. This study evaluated deposits of PM on the leaves and in the wax layer of 35 species (11 shrubs, 24 trees) in Beijing, China. Differences in the accumulation of PM were observed between species. Cephalotaxus sinensis, Euonymus japonicus, Broussonetia papyriferar, Koelreuteria paniculata and Quercus variabilis were all efficient in capturing small particles. The plants exhibiting high amounts of total PM accumulation (on leaf surfaces and/or in the wax layer), also showed comparatively high levels of PM accumulation across all particle sizes. A comparison of shrubs and trees did not reveal obvious differences in their ability to accumulate particles based on growth form; a combination of plantings with different growth forms can efficiently reduce airborne PM concentrations near the ground. To test the relationships between leaf traits and PM accumulation, leaf samples of selected species were observed using a scanning electron microscope. Growth forms with greater amounts of pubescence and increased roughness supported PM accumulation; the adaxial leaf surfaces collected more particles than the abaxial surfaces. The results of this study may inform the selection of species for urban green areas where the goal is to capture air pollutants and mitigate the adverse effects of air pollution on human health.     

Direct demonstration of duvatrienediol biosynthesis in glandular heads of tobacco trichomes

Biosynthesis of the diterpenes, α and β 4,8,13-duvatriene-1,3-diol, has been observed in detached, intact glandular heads from trichomes of Nicotiana tabacum, Tobacco Introduction 1068. This result shows directly that the glandular head portion of the trichome is capable of duvatrienediol biosynthesis. In additional experiments, all of the [¹⁴C] duvatrienediol formed from sodium [2-¹⁴C]acetate by leaf midrib sections was recovered with trichome exudate and surface washes. None was found in trichome stalk, epidermal or subepidermal tissue extracts. Also, removal of glandular heads and exudate from midrib sections reduced or eliminated duvatrienediol biosynthetic capacity. Together these results strongly suggest that glandular heads are the primary, and perhaps the only, site of duvatrienediol biosynthesis in this plant.

Incubation of detached, intact glandular heads with sodium [¹⁴C]acetate in the dark or incubation in the light in the presence of DCMU reduced incorporation into duvatrienediols by 97%. These results suggest that chloroplasts which are abundant in glandular heads are involved in the biogenesis of these compounds.

     

Within-plant distribution of volatile compounds on the leaf surface of Flourensia cernua

We are using Flourensia cernua as a shrub model to study how terpenes affect livestock herbivory. Two experiments were conducted to examine distribution of volatile chemicals within a plant in an effort to minimize sample variability. In Experiment 1, leaves (current year's growth) were collected from 20 tarbush plants. Two leaders were sampled from each of three positions (outer canopy, subcanopy, and basal) in all four quadrants (based on ordinal direction). In Experiment 2, 10 leaders of current year's growth were removed from another 20 plants. Leaders were collected from the outer canopy of each quadrant and separated into thirds before removing leaves, thereby creating three leaf age categories. Volatile compounds were extracted with ethanol and analyzed with gas chromatography/mass spectrometry. Ninety-four chemicals (including 15 unknowns) were present on the leaf surface of F. cernua. Although 14 and 21 compounds differed (P < 0.05) among quadrants in Experiments 1 and 2, respectively, no consistent quadrant effect was detected in either study. Leaf position differed (P < 0.05) for 52 chemicals in Experiment 1 but outer canopy and subcanopy leaves differed for only 10 compounds. In Experiment 2, 63 compounds differed among leaf age categories. Immature leaves contained greater concentrations of 46 chemicals (P < 0.05) than intermediate or mature age categories, but intermediate and mature leaves differed for only seven compounds. Estimated total concentration (i.e., cumulative concentration of all compounds) was not affected by leaf position but varied among leaf age categories (P < 0.05; immature > intermediate > mature). Differences in leaf position were attributed about equally to mono- and sesquiterpenes, whereas leaf age differences were primarily due to sesquiterpenes. Leaf position and age both affect terpene concentration and sampling variability for tarbush. However, little difference was detected between subcanopy and canopy positions. Thus, by avoiding basal sprouts and sampling from the mid-point of current year's growth, sampling variation should be minimal.     

Biomass accumulation and shading effects of epiphytes on leaves of the seagrass, Heterozostera tasmanica, in Victoria,Australia

A method is described for estimating the rate of accumulation of epiphyte biomass on leaves of the seagrass, Heterozostera tasmanica (Martens ex Aschers.) den Hartog and for estimating the effect of epiphyte biomass on photosynthesis of the seagrass. Epiphyte biomass was determined by comparison of the weight per unit area of epiphyte-covered and epiphyte-free leaf blades. Epiphyte weight increased as age of the seagrass leaves increased. Linear regression on epiphyte biomass vs. leaf age estimated the rate of biomass accumulation. Rates varied from 5.7 to 104 μg epiphyte dry weight per cm² of leaf surface per day at three sites in Western Port and Port Phillip Bay, Victoria. Rates of accumulation of epiphyte biomass were generally higher during December through March (summer) than in May (autumn), August (winter) or October (Spring). Light attenuation by epiphytes increase linearly with biomass. The rate of biomass accumulation of epiphytes was compared with leaf growth rate, ambient photon flux density in H. tasmanica beds and the photosynthesis—photon flux density curve of H. tasmanica. This comparison demonstrated that epiphyte biomass can accumulate fast enough to shade H. tasmanica leaves and significantly reduce the time (to less than one half of the leaf life span) in which positive net photosynthesis of the leaf blade is possible.     

Die Wasserbenetzbarkeit von Blattoberflächen und ihre submikroskopische Wachsstruktur

Summary The wettability of a leaf surface is defined by the contact angle between a water-droplet and the surface of the leaf.Contact angles of 60–80° were measured on easily wettable leaves. These leaves have no wax on the outer cuticular layer.Contact angles of 130–160° were measured on leaves with a low wettability. These leaves have wax on the outer cuticular layer, which shows submicroscopic structures characteristic of the particular plants.A comparison of the wettability and the different structures of the wax showed no true distinctions.The wax does not adhere strongly to the outer cuticular layer and is cast off in the case of old leaves. Therefore the wetability alters with the age of the leaves. If the wax on young leaves is destroyed or removed by outer influences it can be produced again within a few hours.The rebuilt structure of the wax is not always similar to the original one. When the wax was dissolved by organic solvents no new wax formation was observed.Cigarette smoke greatly increases the wettability of leaves of Tropaeolum majus, parochetus communis and Chelidonium majus without causing a visible alteration of the submicroscopic structure. In this case the leaves recover their water-repellency within a few hours, but not if they were treated with Diesel smoke.When the wax structure was destroyed by fungus or Aleurodina no new wax formation was observed.

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Herrn Professor Dr. Walter Rentschler zum 60. Geburtstag