Stomatal Responses to Flooding of the Intercellular Air Spaces Suggest a Vapor-Phase Signal Between the Mesophyll and the Guard Cells1[OA

Science.gov (United States)

Sibbernsen, Erik; Mott, Keith A.

2010-01-01

Flooding the intercellular air spaces of leaves with water was shown to cause rapid closure of stomata in Tradescantia pallida, Lactuca serriola, Helianthus annuus, and Oenothera caespitosa. The response occurred when water was injected into the intercellular spaces, vacuum infiltrated into the intercellular spaces, or forced into the intercellular spaces by pressurizing the xylem. Injecting 50 mm KCl or silicone oil into the intercellular spaces also caused stomata to close, but the response was slower than with distilled water. Epidermis-mesophyll grafts for T. pallida were created by placing the epidermis of one leaf onto the exposed mesophyll of another leaf. Stomata in these grafts opened under light but closed rapidly when water was allowed to wick between epidermis and the mesophyll. When epidermis-mesophyll grafts were constructed with a thin hydrophobic filter between the mesophyll and epidermis stomata responded normally to light and CO2. These data, when taken together, suggest that the effect of water on stomata is caused partly by dilution of K+ in the guard cell and partly by the existence of a vapor-phase signal that originates in the mesophyll and causes stomata to open in the light. PMID:20472750

A new mechanism for the regulation of stomatal aperture size in intact leaves: accumulation of mesophyll-derived sucrose in the guard-cell wall of Vicia faba

International Nuclear Information System (INIS)

Lu, P.; Outlaw, W.H. Jr.; Smith, B.G.; Freed, G.A.

1997-01-01

At various times after pulse-labeling broad bean (Vicia faba L.) leaflets with 14CO2, whole-leaf pieces and rinsed epidermal peels were harvested and subsequently processed for histochemical analysis. Cells dissected from whole leaf retained apoplastic contents, whereas those from rinsed peels contained only symplastic contents. Sucrose (Suc)-specific radioactivity peaked (111 GBq mol-1) in palisade cells at 20 min. In contrast, the 14C content and Suc-specific radioactivity were very low in guard cells for 20 min, implying little CO2 incorporation; both then peaked at 40 min. The guard-cell apoplast had a high maximum Suc-specific radioactivity (204 GBq mol-1) and a high Suc influx rate (0.05 pmol stoma-1 min-1). These and other comparisons implied the presence of (a) multiple Suc pools in mesophyll cells, (b) a localized mesophyll-apoplast region that exchanges with phloem and stomata, and (c) mesophyll-derived Suc in guard-cell walls sufficient to diminish stomatal opening by approximately 3 micrometers. Factors expected to enhance Suc accumulation in guard-cell walls are (a) high transpiration rate, which closes stomata, and (b) high apoplastic Suc concentration, which is elevated when mesophyll Suc efflux exceeds translocation. Therefore, multiple physiological factors are integrated in the attenuation of stomatal aperture size by this previously unrecognized mechanism

Intracellular compartimentation of abscisic acid (ABA) in guard cells and mesophyll cells under exposure to SO sub 2. Kompartimentierung von Abscisinsaeure (ABA) in Schliess- und Mesophyllzellen unter SO sub 2 -Belastung

Energy Technology Data Exchange (ETDEWEB)

Baier, M.; Daeter, W.; Hartung, W. (Wuerzburg Univ. (Germany, F.R.). Lehrstuhl fuer Botanik 1)

1989-07-01

The effect of SO{sub 2} on the intracellular compartimentation of ABA in guard cells and mesophyll cells of Valerianella locusta was investigated, using the efflux compartmental analysis, as described by Behl and Hartung (1986). The cytoplasmic ABA content of the guard cells was reduced drastically by 6 {mu}molxm{sup -3} SO{sub 2} (20% of the controls). The vacuolar content was decreased less dramatically (70% of the controls). The ABA distribution of mesophyll cells remained uneffected by 6 {mu}molxm{sup -3} SO{sub 2}. The SO{sub 2} effects are explained by an acidification of the compartments. (orig.).

Carbon isotope ratios of epidermal and mesophyll tissues from leaves of C3 and CAM plants

International Nuclear Information System (INIS)

Nishida, K.; Roksandic, Z.; Osmond, B.

1981-01-01

The δ 13 C values for epidermal and mesophyll tissues of two C 3 plants, Commelina communis and Tulipa gesneriana, and a CAM plant, Kalanchoē daigremontiana, were measured. The values for the tissues of both C 3 plants were similar. In young leaves of Kalanchoē, the epidermis and the mesophyll showed S 13 C values which were nearly identical, and similar to those found in C 3 plants. However, markedly more negative values for epidermal compared to mesophyll tissue, were obtained in the mature Kalanchoē leaf. This is consistent with the facts that the epidermis in a CAM leaf is formed when leaves engage in C 3 photosynthesis and that subsequent dark CO 2 fixation in guard cells or mesophyll cells makes only a small contribution to total epidermal carbon

Kinetics of Ca2+- and ATP-dependent, voltage-controlled anion conductance in the plasma membrane of mesophyll cells of Pisum sativum

NARCIS (Netherlands)

Elzenga, J.T.M.; van Volkenburgh, E.

Whole-cell patch-clamp techniques were used to measure anion currents through the plasma membrane of protoplasts of mesophyll cells of expanding pea (Pisum sativum L.) leaves. Voltage-induced changes of the currents could be modelled with single exponential activation and deactivation kinetics. The

Structural characterization of a mixed-linkage glucan deficient mutant reveals alteration in cellulose microfibril orientation in rice coleoptile mesophyll cell walls

Directory of Open Access Journals (Sweden)

Andreia Michelle Smith-Moritz

2015-08-01

Full Text Available The CELLULOSE SYNTHASE-LIKE F6 (CslF6 gene was previously shown to mediate the biosynthesis of mixed-linkage glucan (MLG, a cell wall polysaccharide that is hypothesized to be a tightly associated with cellulose and also have a role in cell expansion in the primary cell wall of young seedlings in grass species. We have recently shown that loss-of-function cslf6 rice mutants do not accumulate MLG in most vegetative tissues. Despite the absence of a structurally important polymer, MLG, these mutants are unexpectedly viable and only show a moderate growth compromise compared to wild type. Therefore these mutants are ideal biological systems to test the current grass cell wall model. In order to gain a better understanding of the role of MLG in the primary wall, we performed in-depth compositional and structural analyses of the cell walls of three day-old rice seedlings using various biochemical and novel microspectroscopic approaches. We found that cellulose content as well as matrix polysaccharide composition was not significantly altered in the MLG deficient mutant. However, we observed a significant change in cellulose microfibril bundle organization in mesophyll cell walls of the cslf6 mutant. Using synchrotron source Fourier Transform Mid-Infrared Spectromicroscopy for high-resolution imaging, we determined that the bonds associated with cellulose and arabinoxylan, another major component of the primary cell was of grasses, were in a lower energy configuration compared to wild type, suggesting a slightly weaker primary wall in MLG deficient mesophyll cells. Taken together, these results suggest that MLG may influence cellulose deposition in mesophyll cell walls without significantly affecting anisotropic growth thus challenging MLG importance in cell wall expansion.

Effects of water stress on photosynthetic electron transport, photophosphorylation, and metabolite levels of Xanthium strumarium mesophyll cells.

Science.gov (United States)

Sharkey, T D; Badger, M R

1982-12-01

Several component processes of photosynthesis were measured in osmotically stressed mesophyll cells of Xanthium strumarium L. The ribulose-1,5-bisphosphate regeneration capacity was reduced by water stress. Photophoshorylation was sensitive to water stress but photosynthetic electron transport was unaffected by water potentials down to-40 bar (-4 MPa). The concentrations of several intermediates of the photosynthetic carbon-reduction cycle remained relatively constant and did not indicate that ATP supply was limiting photosynthesis in the water-stressed cells.

Plant, cell, and molecular mechanisms of abscisic-acid regulation of stomatal apertures. A new mechanism for the regulation of stomatal-aperture size in intact leaves: Accumulation of mesophyll-derived sucrose in the guard-cell wall of Vicia faba L.

Energy Technology Data Exchange (ETDEWEB)

Lu, P.; Outlaw, W.H. Jr.; Smith, B.G.; Freed, G.A.

1996-12-31

At various times after pulse labeling Vicia faba L. leaflets with {sup 14}CO{sub 2}, whole-leaf pieces and rinsed epidermal peels were harvested and subsequently processed for histochemical analysis. Cells dissected from whole leaf retained apoplastic contents whereas those from rinsed peels contained only cytoplastic contents. Sucrose specific radioactivity peaked in palisade cells, 111 GBq{center_dot}mol{sup {minus}1}, at 20 min. In contrast, the {sup 14}C content and sucrose specific radioactivity were very low in guard cells for 20 min, implying little CO{sub 2} incorporation; both then peaked at 40 min. The guard-cell apoplast had a high maximum sucrose specific radioactivity and a high sucrose influx rate. These and other comparisons implied the presence of (a) multiple sucrose pools in mesophyll cells, (b) a localized mesophyll-apoplast region that exchanges with phloem and stomata, and (c) mesophyll-derived sucrose in guard-cell walls sufficient to diminish stomatal opening by {approximately} 4 {micro}m. Factors expected to enhance sucrose accumulation in guard-cell walls are (a) high transpiration rate, which closes stomata, and (b) high apoplastic sucrose concentration, which is elevated when mesophyll-sucrose efflux exceeds translocation. Therefore, multiple physiological factors are integrated in the attenuation of stomatal-aperture size by this previously unrecognized mechanism.

Identification of the TaBTF3 gene in wheat (Triticum aestivum L.) and the effect of its silencing on wheat chloroplast, mitochondria and mesophyll cell development.

Science.gov (United States)

Ma, Hong-Zhen; Liu, Guo-Qin; Li, Cheng-Wei; Kang, Guo-Zhang; Guo, Tian-Cai

2012-10-05

The full-length cDNA (882bp) and DNA (1742bp) sequences encoding a basic transcription factor 3, designated as TaBTF3, were first isolated from common wheat (Triticum aestivum L.). Subcellular localization studies revealed that the TaBTF3 protein was mainly located in the cytoplasm and nucleus. In TaBTF3-silenced transgenic wheat seedlings obtained using the Virus-induced gene silencing (VIGS) method, the chlorophyll pigment content was markedly reduced. However, the malonaldehyde (MDA) and H(2)O(2) contents were enhanced, and the structure of the wheat mesophyll cell was seriously damaged. Furthermore, transcripts of the chloroplast- and mitochondrial-encoded genes were significantly reduced in TaBTF3-silenced transgenic wheat plants. These results suggest that the TaBTF3 gene might function in the development of the wheat chloroplast, mitochondria and mesophyll cell. This paper is the first report to describe the involvement of TaBTF3 in maintaining the normal plant mesophyll cell structure. Copyright © 2012 Elsevier Inc. All rights reserved.

Early H2O2 Accumulation in Mesophyll Cells Leads to Induction of Glutathione during the Hyper-Sensitive Response in the Barley-Powdery Mildew Interaction1

Science.gov (United States)

Vanacker, Helene; Carver, Tim L.W.; Foyer, Christine H.

2000-01-01

H2O2 production and changes in glutathione, catalase, and peroxidase were followed in whole-leaf extracts from the susceptible (AlgS [Algerian/4* (F14) Man.(S)]; ml-a1 allele) and resistant (AlgR [Algerian/4* (F14) Man.(R)]; Ml-a1 allele) barley (Hordeum vulgare) isolines between 12 and 24 h after inoculation with powdery mildew (Blumeria graminis [DC]. Speer [syn. Erysiphe graminis DC] f.sp hordei Marchal). Localized papilla responses and cell death hypersensitive responses were not observed within the same cell. In hypersensitive response sites, H2O2 accumulation first occurred in the mesophyll underlying the attacked epidermal cell. Subsequently, H2O2 disappeared from the mesophyll and accumulated around attacked epidermal cells. In AlgR, transient glutathione oxidation coincided with H2O2 accumulation in the mesophyll. Subsequently, total foliar glutathione and catalase activities transiently increased in AlgR. These changes, absent from AlgS, preceded inoculation-dependent increases in peroxidase activity that were observed in both AlgR and AlgS at 18 h. An early intercellular signal precedes H2O2, and this elicits anti-oxidant responses in leaves prior to events leading to death of attacked cells. PMID:10938348

The influence of nitric oxide and mercury chloride on leaf mesophyll structure under natural drought conditions

Directory of Open Access Journals (Sweden)

Mykola M. Musiyenko

2012-03-01

Full Text Available It is established that under natural drought conditions starch was accumulated in the central part of chloroplasts of mesophyll cells and chloroplasts were localized on the periphery of cells at plasmalemma. After treatment wheat plants by nitric oxide donor the decreasing of starch deposits number and close contacts between chloroplasts were indicated, elongated nucleus was localized in the centre of cells. After treatment wheat plant by mercury chloride chloroplasts in the cells lost their oval shape and contacts, increased eventually deposition of starch, indicating the acceleration of aging tissues. Thus, nitric oxide in drought conditions reduced the destructive effect of drought on mesophyll cells, and mercury chloride caused deformation of the membrane cell.

American wild celery (Vallisneria americana): Ecological considerations for restoration

Science.gov (United States)

Korschgen, C.E.; Green, W.L.

1988-01-01

The success of vegetation management programs for waterfowl is dependent on knowing the physical and physiological requirements of target species. Lakes and riverine impoundments that contain an abundance of the American wildcelery (Vallisneria americana ) have traditionally been favored by canvasbacks (Aythya valisineria ) and other waterfowl as feeding areas during migration. Information on the ecology of American wildcelery is summarized to serve as a guide for potential wetland restoration projects. Techniques are described for transplanting winter buds. Management programs that employ these techniques should define objectives clearly and evaluate the water regime carefully before initiating major restoration.

Biosorption of arsenic from groundwater using Vallisneria gigantea plants. Kinetics, equilibrium and photophysical considerations.

Science.gov (United States)

Iriel, Analia; Lagorio, M Gabriela; Fernández Cirelli, Alicia

2015-11-01

Arsenic (V) uptake from groundwater by using Vallisneria gigantea plants was studied using batch experiments. Reflectance and fluorescence of intact plants were investigated and changes in photophysical properties following arsenic absorption were reported. Good correlations have been found between arsenic concentration in groundwater and parameters derived from reflectance and fluorescence measurements. This system reached its equilibrium after seven days when the removal quantities were strongly dependent on the initial arsenic concentration. Interestingly, Vallisneria plants were able to accumulate from 100 to 600 mg As kg(-1) in roots and fronds although the translocation factors were low (0.6-1.6). Kinetic data for biosorption process followed a first-order law. At low arsenic concentrations the uptake in plants was governed by diffusion aspects. Langmuir, Freundlich and Dubinin-Radushkevich models were applied and results demonstrated that arsenic uptake was better described by the Langmuir model. As a final remark we concluded that a plant of this species should be able to remove 1mg As per week. Copyright © 2015 Elsevier Ltd. All rights reserved.

Macroautophagy and microautophagy in relation to vacuole formation in mesophyll cells of Dendrobium tepals.

Science.gov (United States)

van Doorn, Wouter G; Kirasak, Kanjana; Ketsa, Saichol

2015-04-01

Prior to flower opening, mesophyll cells at the vascular bundles of Dendrobium tepals showed a large increase in vacuolar volume, partially at the expense of the cytoplasm. Electron micrographs indicated that this increase in vacuolar volume was mainly due to vacuole fusion. Macroautophagous structures typical of plant cells were observed. Only a small part of the decrease in cytoplasmic volume seemed due to macroautophagy. The vacuoles contained vesicles of various types, including multilamellar bodies. It was not clear if these vacuolar inclusions were due to macroautophagy or microautophagy. Only a single structure was observed of a protruding vacuole, indicating microautophagy. It is concluded that macroautophagy occurs in these cells but its role in vacuole formation seems small, while a possible role of microautophagy in vacuole formation might be hypothesized. Careful labeling of organelle membranes seems required to advance our insight in plant macro- and microautophagy and their roles in vacuole formation. Copyright © 2015 Elsevier GmbH. All rights reserved.

Influence of pH on the /sup 14/C-labelling pattern after photosynthesis of suspended leaf slices and isolated mesophyll cells from chenopodium album in NaH/sup 14/CO/sub 3/

Energy Technology Data Exchange (ETDEWEB)

Baumann, G; Guenther, G [Paedagogische Hochschule Karl Liebknecht, Potsdam (German Democratic Republic). Sektion Chemie/Biologie

1983-01-01

Photosynthetic fixation of /sup 14/C from solutions of NaH/sup 14/CO/sub 3/ (at constant concentrations of free CO/sub 2/) by suspended leaf slices or isolated mesophyll cells from Chenopodium album is increased with increasing pH. Above all, the incorporation of radioactivity into amino acids and malate is stimulated. A direct uptake of HCO/sub 3/ ions and its fixation by PEP carboxylase is suggested. Isolated mesophyll cells showed at pH 7.3 a higher rate of photosynthesis than at pH 5.0.

Effects of shading on Vallisneria natans (Lour.) H. Hara growth

DEFF Research Database (Denmark)

Fox, Anthony David; Meng, F; Shen, X

2013-01-01

Effects of surface shading were measured on above- and below-ground biomass and fruit production of Vallisneria natans (Lour.) H. Hara plants grown from seed in replicated microcosm experiments, based on a control (no shading) and four treatments (25%, 50%, 75% and 90% shading). Above- and below-...... spp. at seasonally inundated wetlands in the Yangtze River floodplain could, by shading, have contributed to the reduction in annual biomass and seed production of V. natans, contributing to declines in distribution and abundance....

Estimation of mesophyll conductance to CO2 flux by three different methods

International Nuclear Information System (INIS)

Loreto, F.; Harley, P.C.; Di Marco, G.; Sharkey, T.D.

1992-01-01

The resistance to diffusion of CO2 from the intercellular airspaces within the leaf through the mesophyll to the sites of carboxylation during photosynthesis was measured using three different techniques, The three techniques include a method based on discrimination against the heavy stable isotope of carbon, 13C, and two modeling methods. The methods rely upon different assumptions, but the estimates of mesophyll conductance were similar with all three methods. The mesophyll conductance of leaves from a number of species was about 1.4 times the stomatal conductance for CO2 diffusion determined in unstressed plants at high light. The relatively low CO2 partial pressure inside chloroplasts of plants with a low mesophyll conductance did not lead to enhanced O2 sensitivity of photosynthesis because the low conductance caused a significant drop in the chloroplast CO2 Partial pressure upon switching to low O2. We found no correlation between mesophyll conductance and the ratio of internal leaf area to leaf surface area and only a weak correlation between mesophyll conductance and the proportion of leaf volume occupied by air. Mesophyll conductance was independent of CO2 and O2 partial pressure during the measurement, indicating that a true physical parameter, independent of biochemical effects, was being measured. No evidence for accumulating mechanisms was found. Some plants, notably Citrus aurantium and Simmondsia chinensis, had very low conductances that limit the rate of photosynthesis these plants can attain at atmospheric CO2 level

Stomatal conductance, mesophyll conductance, and transpiration efficiency in relation to leaf anatomy in rice and wheat genotypes under drought.

Science.gov (United States)

Ouyang, Wenjing; Struik, Paul C; Yin, Xinyou; Yang, Jianchang

2017-11-02

Increasing leaf transpiration efficiency (TE) may provide leads for growing rice like dryland cereals such as wheat (Triticum aestivum). To explore avenues for improving TE in rice, variations in stomatal conductance (gs) and mesophyll conductance (gm) and their anatomical determinants were evaluated in two cultivars from each of lowland, aerobic, and upland groups of Oryza sativa, one cultivar of O. glaberrima, and two cultivars of T. aestivum, under three water regimes. The TE of upland rice, O. glaberrima, and wheat was more responsive to the gm/gs ratio than that of lowland and aerobic rice. Overall, the explanatory power of the particular anatomical trait varied among species. Low stomatal density mostly explained the low gs in drought-tolerant rice, whereas rice genotypes with smaller stomata generally responded more strongly to drought. Compared with rice, wheat had a higher gm, which was associated with thicker mesophyll tissue, mesophyll and chloroplasts more exposed to intercellular spaces, and thinner cell walls. Upland rice, O. glaberrima, and wheat cultivars minimized the decrease in gm under drought by maintaining high ratios of chloroplasts to exposed mesophyll cell walls. Rice TE could be improved by increasing the gm/gs ratio via modifying anatomical traits. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Influence of Sulfur on the Arsenic Phytoremediation Using Vallisneria natans (Lour.) Hara.

Science.gov (United States)

Chen, Guoliang; Feng, Tao; Li, Zhixian; Chen, Zhang; Chen, Yuanqi; Wang, Haihua; Xiang, Yanci

2017-09-01

Influences of sulfur (S) on the accumulation and detoxification of arsenic (As) in Vallisneria natans (Lour.) Hara, an arsenic hyperaccumulating submerged aquatic plant, were investigated. At low sulfur levels (75%) present in the plant after exposure to As(V). Sulfur plays an important role in the arsenic translocation and detoxification, possibly through stimulating the synthesis of thiols and complexation of arsenite-phytochelatins. This suggests that addition of sulfur to the arsenic-contaminated water may provide a way to promote arsenic bioaccumulation in plants for phytoremediation of arsenic pollution.

Effects of herbicides on /sup 14/CO/sub 2/ fixation in isolated mesophyll cells from Beta vulgaris (sugar beet) and Chenopodium album

Energy Technology Data Exchange (ETDEWEB)

Baumann, G; Guenther, G [Paedagogische Hochschule Karl Liebknecht, Potsdam (German Democratic Republic)

1979-01-01

10/sup -4/ - 10/sup -6/ molar solutions of herbicides (atrazine, 2,4-D, desmetryne, diallate, diquat, feuron, lenacil, NaTa, paraquat, phenmedipham, prometryne, propham, pyrazone, and simazine) cause similar inhibitory effects on the photosynthetic /sup 14/CO/sub 2/ fixation in isolated mesophyll cells from Chenopodium album and Beta vulgaris. Correlatdion between inhibition and herbicide resistance of the whole plants could be realized for lenacil only.

[Effects of light intensities after anthesis on the photosynthetic characteristics and chloroplast ultrastructure in mesophyll cell of summer maize (Zea mays L. )].

Science.gov (United States)

Gao, Jia; Cui, Hai Yan; Shi, Jian Guo; Dong, Shu Ting; Liu, Peng; Zhao, Bin; Zhang, Ji Wang

2018-03-01

We examined the changes of photosynthetic characteristics and chloroplast ultrastructure in mesophyll cell of summer maize in response to different light intensities in the field, with the summer maize hybrid Denghai 605 as experimental material. Two treatments of both shading (S) and increasing light (L) from flowering to physiological maturity stage were designed, with the ambient sunlight treatment as control (CK). Under shading treatment, poorly developed thylakoid structure, blurry lamellar structure, loose granum, large gap between slices and warping granum were the major characteristics in chloroplast. Meanwhile, photosynthetic rate (P n ), transpiration rate, stomatal conductance, chlorophyll content, and actual photo-chemical efficiency (Φ PSII ) decreased, whereas the maximal photochemical efficiency and non-photochemical quenching increased, which resulted in decreases in grain yield under shading treatment. However, a better development was observed in chloroplasts for L treatment, with the number of grana and lamellae increased and lamellae arranged compactly. In addition, P n and Φ PSII increased under L treatment, which increased grain yield. The chloroplast arrangement dispersed in mesophyll cells and chloroplast ultrastructure was destroyed after shading, and then chlorophyll synthesis per unit leaf area and photosynthetic capacity decreased. In contrast, the number of grana and lamellae increased and lamellae arranged compactly after increasing light, which are beneficial for corn yield.

Disruption of stomatal lineage signaling or transcriptional regulators has differential effects on mesophyll development, but maintains coordination of gas exchange.

Science.gov (United States)

Dow, Graham J; Berry, Joseph A; Bergmann, Dominique C

2017-10-01

Stomata are simultaneously tasked with permitting the uptake of carbon dioxide for photosynthesis while limiting water loss from the plant. This process is mainly regulated by guard cell control of the stomatal aperture, but recent advancements have highlighted the importance of several genes that control stomatal development. Using targeted genetic manipulations of the stomatal lineage and a combination of gas exchange and microscopy techniques, we show that changes in stomatal development of the epidermal layer lead to coupled changes in the underlying mesophyll tissues. This coordinated response tends to match leaf photosynthetic potential (V cmax ) with gas-exchange capacity (g smax ), and hence the uptake of carbon dioxide for water lost. We found that different genetic regulators systematically altered tissue coordination in separate ways: the transcription factor SPEECHLESS (SPCH) primarily affected leaf size and thickness, whereas peptides in the EPIDERMAL PATTERNING FACTOR (EPF) family altered cell density in the mesophyll. It was also determined that interlayer coordination required the cell-surface receptor TOO MANY MOUTHS (TMM). These results demonstrate that stomata-specific regulators can alter mesophyll properties, which provides insight into how molecular pathways can organize leaf tissues to coordinate gas exchange and suggests new strategies for improving plant water-use efficiency. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

Impact of human activities on water level and clarity and underwater light climate of Vallisneria spiralis L. in Poyan Lake, China

NARCIS (Netherlands)

Wu, G.

2008-01-01

Almost 95% of the world population of Siberian crane (Grus leucogeranus) winter in Poyang Lake, China. Here they forage on the tubers of the submerged aquatic macrophyte Vallisneria spiralis L. The growth and production of V. spiralis are regulated by the local hydrology, which might also be

Increasing Leaf Vein Density via Mutagenesis in Rice Results in an Enhanced Rate of Photosynthesis, Smaller Cell Sizes and Can Reduce Interveinal Mesophyll Cell Number

Directory of Open Access Journals (Sweden)

Aryo B. Feldman

2017-11-01

Full Text Available Improvements to leaf photosynthetic rates of crops can be achieved by targeted manipulation of individual component processes, such as the activity and properties of RuBisCO or photoprotection. This study shows that simple forward genetic screens of mutant populations can also be used to rapidly generate photosynthesis variants that are useful for breeding. Increasing leaf vein density (concentration of vascular tissue per unit leaf area has important implications for plant hydraulic properties and assimilate transport. It was an important step to improving photosynthetic rates in the evolution of both C3 and C4 species and is a foundation or prerequisite trait for C4 engineering in crops like rice (Oryza sativa. A previous high throughput screen identified five mutant rice lines (cv. IR64 with increased vein densities and associated narrower leaf widths (Feldman et al., 2014. Here, these high vein density rice variants were analyzed for properties related to photosynthesis. Two lines were identified as having significantly reduced mesophyll to bundle sheath cell number ratios. All five lines had 20% higher light saturated photosynthetic capacity per unit leaf area, higher maximum carboxylation rates, dark respiration rates and electron transport capacities. This was associated with no significant differences in leaf thickness, stomatal conductance or CO2 compensation point between mutants and the wild-type. The enhanced photosynthetic rate in these lines may be a result of increased RuBisCO and electron transport component amount and/or activity and/or enhanced transport of photoassimilates. We conclude that high vein density (associated with altered mesophyll cell length and number is a trait that may confer increased photosynthetic efficiency without increased transpiration.

Cell wall matrix polysaccharide distribution and cortical microtubule organization: two factors controlling mesophyll cell morphogenesis in land plants.

Science.gov (United States)

Sotiriou, P; Giannoutsou, E; Panteris, E; Apostolakos, P; Galatis, B

2016-03-01

This work investigates the involvement of local differentiation of cell wall matrix polysaccharides and the role of microtubules in the morphogenesis of mesophyll cells (MCs) of three types (lobed, branched and palisade) in the dicotyledon Vigna sinensis and the fern Asplenium nidus. Homogalacturonan (HGA) epitopes recognized by the 2F4, JIM5 and JIM7 antibodies and callose were immunolocalized in hand-made leaf sections. Callose was also stained with aniline blue. We studied microtubule organization by tubulin immunofluorescence and transmission electron microscopy. In both plants, the matrix cell wall polysaccharide distribution underwent definite changes during MC differentiation. Callose constantly defined the sites of MC contacts. The 2F4 HGA epitope in V. sinensis first appeared in MC contacts but gradually moved towards the cell wall regions facing the intercellular spaces, while in A. nidus it was initially localized at the cell walls delimiting the intercellular spaces, but finally shifted to MC contacts. In V. sinensis, the JIM5 and JIM7 HGA epitopes initially marked the cell walls delimiting the intercellular spaces and gradually shifted in MC contacts, while in A. nidus they constantly enriched MC contacts. In all MC types examined, the cortical microtubules played a crucial role in their morphogenesis. In particular, in palisade MCs, cortical microtubule helices, by controlling cellulose microfibril orientation, forced these MCs to acquire a truncated cone-like shape. Unexpectedly in V. sinensis, the differentiation of colchicine-affected MCs deviated completely, since they developed a cell wall ingrowth labyrinth, becoming transfer-like cells. The results of this work and previous studies on Zea mays (Giannoutsou et al., Annals of Botany 2013; 112: : 1067-1081) revealed highly controlled local cell wall matrix differentiation in MCs of species belonging to different plant groups. This, in coordination with microtubule-dependent cellulose microfibril

Will the Three Gorges Dam affect the underwater light climate of Vallisneria spiralis L. and food habitat of Siberian crane in Poyang Lake?

NARCIS (Netherlands)

Wu, G.; Leeuw, de J.; Skidmore, A.K.; Prins, H.H.T.; Best, E.P.H.; Liu, Y.

2009-01-01

Almost 95% of the entire population of the Siberian crane (Grus leucogeranus) winter in Poyang Lake, China, where they forage on the tubers of the submerged aquatic macrophyte Vallisneria spiralis. The Three Gorges Dam on the Yangtze River may possibly affect this food source of the Siberian crane

Ultrastructural response of cabbage outer leaf mesophyll cells (Brassica oleracea L. to excess of nickel

Directory of Open Access Journals (Sweden)

Jolanta Molas

2014-01-01

Full Text Available Changes in the structure and in the ultrastructure of cabbage outer leaf mesophyll cells [Brassica oleracea L.] cv. Sława from Enkhouizen were examined by means of light and electron microscopy. The examined plants were grown on the basic Murashige and Skoog medium with addition of excesive concentrations of nickel (added as NiSO4 x 7H2O,i.e. Ni 5, Ni 10 and Ni 20 mg/dm3. In Ni 5 mg samples mainly adaptation changes to the conditions of stress were observed. These changes were manifested by the increase of cytoplasm content and by cytoplasm vacuolization, by the increase of nucleus and nucleous volume, nucleolus vacuolization, the increase of plasmalemma invaginations and of the amount of rough ER, by the central arrangement of smooth ER and of the thylakoids of chloroplasts; it was also shown by the growth of the number of mitochondria and of peroxisomes in the cell. In Ni 10 mg samples, apart from adaptation changes, such as the increase of the nucleus volume, increase of plasmalemma invaginations, cytoplasm and nucleolus vacuolization, degeneration changes were also observed. They concerned mainly the nucleus (the increasing amount of condensed chromatin, ER (swelling and fragmentation of rER and sER, mitochondrium (swelling and reduction of cristae, Golgi apparatus (disintegration and decay and chloroplasts (changes of shape, swelling and reduction of thylakoids, disappearance of starch and presence of big plastoglobuli. In Ni 20 mg samples cell protoplasts were in different stages of degeneration and the cell organelles that were identifiable, were usually damaged.

Early local differentiation of the cell wall matrix defines the contact sites in lobed mesophyll cells of Zea mays.

Science.gov (United States)

Giannoutsou, E; Sotiriou, P; Apostolakos, P; Galatis, B

2013-10-01

The morphogenesis of lobed mesophyll cells (MCs) is highly controlled and coupled with intercellular space formation. Cortical microtubule rings define the number and the position of MC isthmi. This work investigated early events of MC morphogenesis, especially the mechanism defining the position of contacts between MCs. The distributions of plasmodesmata, the hemicelluloses callose and (1 → 3,1 → 4)-β-d-glucans (MLGs) and the pectin epitopes recognized by the 2F4, JIM5, JIM7 and LM6 antibodies were studied in the cell walls of Zea mays MCs. Matrix cell wall polysaccharides were immunolocalized in hand-made sections and in sections of material embedded in LR White resin. Callose was also localized using aniline blue in hand-made sections. Plasmodesmata distribution was examined by transmission electron microscopy. Before reorganization of the dispersed cortical microtubules into microtubule rings, particular bands of the longitudinal MC walls, where the MC contacts will form, locally differentiate by selective (1) deposition of callose and the pectin epitopes recognized by the 2F4, LM6, JIM5 and JIM7 antibodies, (2) degradation of MLGs and (3) formation of secondary plasmodesmata clusterings. This cell wall matrix differentiation persists in cell contacts of mature MCs. Simultaneously, the wall bands between those of future cell contacts differentiate with (1) deposition of local cell wall thickenings including cellulose microfibrils, (2) preferential presence of MLGs, (3) absence of callose and (4) transient presence of the pectins identified by the JIM5 and JIM7 antibodies. The wall areas between cell contacts expand determinately to form the cell isthmi and the cell lobes. The morphogenesis of lobed MCs is characterized by the early patterned differentiation of two distinct cell wall subdomains, defining the sites of the future MC contacts and of the future MC isthmi respectively. This patterned cell wall differentiation precedes cortical microtubule

Effects of shading on Vallisneria natans (Lour. H. Hara growth

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Fox A.D.

2013-08-01

Full Text Available Effects of surface shading were measured on above- and below-ground biomass and fruit production of Vallisneria natans (Lour. H. Hara plants grown from seed in replicated microcosm experiments, based on a control (no shading and four treatments (25%, 50%, 75% and 90% shading. Above- and below-ground biomass was significantly reduced at treatments above 50% shading and first pistillate and staminate florescence dates were significantly delayed above 75% and 50% shading, respectively. Ratios of mature to unripe fruits produced (both in number or dry weight did not differ between shading treatments, but dry weight fruit production was significantly reduced at 90% shading. We conclude that above 50% surface shading, V. natans plants suffer reductions in accumulated biomass and investment in sexual reproduction. We contend that recent expansions in the extent of the native floating water chestnut Trapa spp. at seasonally inundated wetlands in the Yangtze River floodplain could, by shading, have contributed to the reduction in annual biomass and seed production of V. natans, contributing to declines in distribution and abundance.

Interspecific competition effects on phosphorus accumulation by Hydrilla verticillata and Vallisneria natans.

Science.gov (United States)

Zhang, Xiufeng; Liu, Zhengwen

2011-01-01

The competition between submersed plants has been recognized as an important factor influencing the structure of plant communities in shallow lakes. The ability of different species to take up and store nutrients from the surrounding ambience varies, and hence plant community structure might be expected to affect the cycling of nutrients in lake ecosystems. In this study, the uptake of phosphorus by Hydrilla verticillata and Vallisneria natans was studied and compared in monoculture and competitive mixed-culture plantings. Results showed that for both studied species the phosphorus concentrations of different tissues and of whole plants was unaffected by competition. However, the quantity of phosphorus accumulated by whole plants of H. verticillata was significantly higher in mixture culture than in monoculture, while that of V. natans was lower in the mixed culture. The results indicated that H. verticillata has a competitive advantage over V. natans, when the two species are grown in competition, and is able to accumulate a greater quantity of phosphorus.

Differential gene expression and transport functionality in the bundle sheath versus mesophyll - a potential role in leaf mineral homeostasis.

Science.gov (United States)

Wigoda, Noa; Pasmanik-Chor, Metsada; Yang, Tianyuan; Yu, Ling; Moshelion, Menachem; Moran, Nava

2017-06-01

Under fluctuating ambient conditions, the ability of plants to maintain hydromineral homeostasis requires the tight control of long distance transport. This includes the control of radial transport within leaves, from veins to mesophyll. The bundle sheath is a structure that tightly wraps around leaf vasculature. It has been suggested to act as a selective barrier in the context of radial transport. This suggestion is based on recent physiological transport assays of bundle sheath cells (BSCs), as well as the anatomy of these cells.We hypothesized that the unique transport functionality of BSCs is apparent in their transcriptome. To test this, we compared the transcriptomes of individually hand-picked protoplasts of GFP-labeled BSCs and non-labeled mesophyll cells (MCs) from the leaves of Arabidopsis thaliana. Of the 90 genes differentially expressed between BSCs and MCs, 45% are membrane related and 20% transport related, a prominent example being the proton pump AHA2. Electrophysiological assays showed that the major AKT2-like membrane K+ conductances of BSCs and MCs had different voltage dependency ranges. Taken together, these differences may cause simultaneous but oppositely directed transmembrane K+ fluxes in BSCs and MCs, in otherwise similar conditions. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Effects of the length of inundation periods on investment in tuber biomass and sexual reproduction by Vallisneria spinulosa S.Z. Yan Ramets

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Fox A.D.

2014-05-01

Full Text Available Modification and reductions in flows are reducing inundation times in some Yangtze River wetlands, potentially affecting the growth of submerged macrophytes. The effects of shortening the inundation period from April to October (control to April to September (treatment were tested on the production of surface and below-ground tubers and fruits in individual Vallisneria spinulosa ramets in microcosm experiments. Mean numbers of below-ground tubers produced did not differ significantly between treatments, but individual tuber dry mass was significantly greater when harvested in October compared with plants harvested in September. Significantly more surface tubers were initiated by the ramets in the later harvest, but these did not differ in dry mass. Fewer ramets had initiated sexual reproduction by the September harvest compared with the October harvest. Almost all October fruits were mature, while the ratio of mature to unripe fruits was significantly greater than in September. There was no support for the hypothesis that more tubers were initiated deeper in the substrate with a longer inundation period to avoid predation by fish or birds. Based on these results, shortening of the inundation periods experienced by Vallisneria spinulosa plants in Yangtze River ephemeral wetlands may reduce the dry biomass of below-ground tubers and the production of surface tubers in this keystone macrophyte.

Effects of potentially acidic air pollutants on the intracellular distribution and transport of plant growth regulators in mesophyll cells of leaves. Consequences on stress- and developmental physiology

Energy Technology Data Exchange (ETDEWEB)

Kremer, H.; Pfanz, H.; Hartung, W.

1987-07-11

The influence of SO/sub 2/ on the intracellular distribution of abscisic acid (ABA) and indole-acetic acid (IAA) in mesophyll cells of Picea abies, Tsuga americana and Hordeum vulgare was investigated. The compartmentation of ABA and IAA depends on intracellular pH-gradients. The hydrophilic anions ABA and IAA are accumulated in the alkaline cell compartments cytosol and chloroplasts, which act as anion traps for weak acids. Uptake of sulfur dioxide into leaves leads to an acidification of alkaline cell compartments, thus decreasing intracellular pH-gradients. Consequently this results in an increased release of plant growth regulators from the cell interior into the apoplast. Therefore the target cells of plant hormones i.e. meristems and stomates are exposed to altered hormone concentrations. Obviously this influences the regulation of cellular metabolism plant development and growth.

Comparative assessment of Azolla pinnata and Vallisneria spiralis in Hg removal from G.B. Pant Sagar of Singrauli Industrial region, India.

Science.gov (United States)

Rai, Prabhat Kumar; Tripathi, B D

2009-01-01

The aim of the present work was to monitor the Hg pollution in water and sediments of G.B. Pant Sagar located in Singrauli Industrial Region, India and to suggest the efficient aquatic plants for its phytoremediation. The study assessed the comparative potential of a free floating water fern Azolla pinnata and submerged aquatic macrophyte Vallisneria spiralis to purify waters polluted by Hg. Six days laboratory experiments have been conducted to mark the percentage removal of Hg at initial concentration of 0.1, 0.5, 1.0 and 3.0 mg L(-1). The percentage removal of Hg was higher for A. pinnata (80-94%) than V. spiralis (70-84%). Likewise, the Hg accumulated in dry mass was much higher for A. pinnata and a high correlation (R(2) = 0.91 for A. pinnata and 0.99 for V. spiralis) was obtained between applied Hg doses and accumulated amounts in biomass. A concentration dependent decrease in chlorophyll a, protein, RNA, DNA and nutrients (NO(3-) and PO(4)(3-)) uptake was detected in A. pinnata and V. spiralis due to Hg toxicity. The decrease was more prominent in Azolla than Vallisneria. The results recommended the use of A. pinnata and V. spiralis to ameliorate the industrial effluents (thermal power, chlor-alkali and coal mine effluent) contaminated with Hg.

Abscisic Acid Induces Rapid Reductions in Mesophyll Conductance to Carbon Dioxide.

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Giuseppe Sorrentino

Full Text Available The rate of photosynthesis (A of plants exposed to water deficit is a function of stomatal (gs and mesophyll (gm conductance determining the availability of CO2 at the site of carboxylation within the chloroplast. Mesophyll conductance often represents the greatest impediment to photosynthetic uptake of CO2, and a crucial determinant of the photosynthetic effects of drought. Abscisic acid (ABA plays a fundamental role in signalling and co-ordination of plant responses to drought; however, the effect of ABA on gm is not well-defined. Rose, cherry, olive and poplar were exposed to exogenous ABA and their leaf gas exchange parameters recorded over a four hour period. Application with ABA induced reductions in values of A, gs and gm in all four species. Reduced gm occurred within one hour of ABA treatment in three of the four analysed species; indicating that the effect of ABA on gm occurs on a shorter timescale than previously considered. These declines in gm values associated with ABA were not the result of physical changes in leaf properties due to altered turgor affecting movement of CO2, or caused by a reduction in the sub-stomatal concentration of CO2 (Ci. Increased [ABA] likely induces biochemical changes in the properties of the interface between the sub-stomatal air-space and mesophyll layer through the actions of cooporins to regulate the transport of CO2. The results of this study provide further evidence that gm is highly responsive to fluctuations in the external environment, and stress signals such as ABA induce co-ordinated modifications of both gs and gm in the regulation of photosynthesis.

Inorganic carbon uptake during photosynthesis. II. Uptake by isolated Asparagus mesophyll cells during isotopic disequilibrium

International Nuclear Information System (INIS)

Espie, G.S.; Owttrim, G.W.; Colman, B.

1986-01-01

The species of inorganic carbon (CO 2 or HCO 3 - ) taken up as a source of substrate for photosynthetic fixation by isolated Asparagus sprengeri mesophyll cells is investigated. Discrimination between CO 2 or HCO 3 - transport, during steady state photosynthesis, is achieved by monitoring the changes (by 14 C fixation) which occur in the specific activity of the intracellular pool of inorganic carbon when the inorganic carbon present in the suspending medium is in a state of isotopic disequilibrium. Quantitative comparisons between theoretical (CO 2 or HCO 3 - transport) and experimental time-courses of 14 C incorporation, over the pH range of 5.2 to 7.5, indicate that the specific activity of extracellular CO 2 , rather than HCO 3 - , is the appropriate predictor of the intracellular specific activity. It is concluded, therefore, that CO 2 is the major source of exogenous inorganic carbon taken up by Asparagus cells. However, at high pH (8.5), a component of net DIC uptake may be attributable to HCO 3 - transport, as the incorporation of 14 C during isotopic disequilibrium exceeds the maximum possible incorporation predicted on the basis of CO 2 uptake alone. The contribution of HCO 3 - to net inorganic carbon uptake (pH 8.5) is variable, ranging from 5 to 16%, but is independent of the extracellular HCO 3 - concentration. The evidence for direct HCO 3 - transport is subject to alternative explanations and must, therefore, be regarded as equivocal. Nonlinear regression analysis of the rate of 14 C incorporation as a function of time indicates the presence of a small extracellular resistance to the diffusion of CO 2 , which is partially alleviated by a high extracellular concentration of HCO 3 -

Guard cell zeaxanthin tracks photosynthetically active radiation and stomatal apertures in Vicia faba leaves

International Nuclear Information System (INIS)

Srivastava, A.; Zeiger, E.

1995-01-01

Zeaxanthin, antheraxanthin and violaxanthin concentrations in guard cells from sonicated abaxial epidermal peels of Vicia faba were measured from dawn to dusk, and compared with concentrations in mesophyll tissue of the same leaves. Measured changes in guard cell zeaxanthin and violaxanthin concentrations indicate that guard cells operate the xanthophyll cycle throughout the day. Mesophyll tissue had no detectable zeaxanthin at dawn, whereas guard cells had 30–50 mmol mol −1 chlorophyll a+b. On a chlorophyll basis, maximal zeaxanthin levels were 3–4 fold higher in guard cells than in mesophyll cells. Zeaxanthin concentrations tracked levels of photosynthetically active radiation (PAR) in both mesophyll and guard cells. In the mesophyll, most of the zeaxanthin changes occurred in mid-morning and mid-afternoon. In guard cells, zeaxanthin concentrations changed nearly linearly with PAR in the early morning and late afternoon, and closely tracked PAR levels throughout the day. Guard cell zeaxanthin concentrations were also closely correlated with stomatal apertures. The close relationship between zeaxanthin concentrations and PAR levels in guard cells indicates that zeaxanthin is well suited to function as a molecular photosensor in stomatal movements. (author)

Infection of potato mesophyll protoplasts with five plant viruses.

Science.gov (United States)

Barker, H; Harrison, B D

1982-12-01

Methods are described for preparing potato mesophyll protoplasts that are suitable for infection with inocula of virus nucleoprotein or RNA. The protoplasts could be infected with four sap-transmissible viruses (tobacco mosaic, tobacco rattle, tobacco ringspot and tomato black ring viruses) and with potato leafroll virus, which is not saptransmissible. No differences were observed in ability to infect protoplasts with potato leafroll virus strains differing either in virulence in intact plants or in aphid transmissibility.

Durum and bread wheat differ in their ability to retain potassium in leaf mesophyll: implications for salinity stress tolerance.

Science.gov (United States)

Wu, Honghong; Shabala, Lana; Zhou, Meixue; Shabala, Sergey

2014-10-01

Understanding the intrinsic mechanisms involved in the differential salinity tolerance between bread wheat and durum wheat is essential for breeding salt-tolerant varieties to cope with the global salinity issue threatening future food supply. In the past, higher salinity tolerance in bread wheat compared with durum wheat has been attributed to its better ability to exclude Na(+) from uptake. Here we show that another mechanism, namely more superior K(+) retention ability in the leaf mesophyll, also contributes to this difference. A strong positive correlation (R(2) > 0.41, P varieties. However, while the above correlation was strong in bread wheat, it was statistically insignificant in durum wheat. Consistent with these findings, a significantly higher relative leaf K(+) content was found in bread wheat than in durum wheat. In contrast to root tissues, the role of voltage-gated K(+) channels in K(+) retention in the wheat mesophyll was relatively small, and non-selective cation channels played a major role in controlling intracellular K(+) homeostasis. Moreover, a significant negative correlation between NaCl-induced mesophyll H(+) flux and mesophyll K(+) retention was found, and interpreted as a compensatory mechanism employed by sensitive varieties to regain K(+) leaked into the apoplast. It is concluded that bread wheat and durum wheat show different strategies of coping with salinity, and that targeting mechanisms conferring K(+) retention in the leaf mesophyll may be a promising way to improve the overall salinity tolerance in these species. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Novel efficient methods for measuring mesophyll anatomical characteristics from fresh thick sections using stereology and confocal microscopy: application on acid rain-treated Norway spruce needles

Czech Academy of Sciences Publication Activity Database

Albrechtová, Jana; Janáček, Jiří; Lhotáková, Zuzana; Radochová, Barbora; Kubínová, Lucie

2007-01-01

Roč. 58, č. 6 (2007), s. 1451-1461 ISSN 0022-0957 R&D Projects: GA AV ČR IAA5011810; GA AV ČR(CZ) IAA600110507; GA MŠk(CZ) LC06063 Institutional research plan: CEZ:AV0Z50110509; CEZ:AV0Z60050516 Keywords : mesophyll * stereology * confocal microscopy Subject RIV: EA - Cell Biology Impact factor: 3.917, year: 2007

Phytotoxicity of four herbicides on Ceratophyllum demersum, Vallisneria natans and Elodea nuttallii.

Science.gov (United States)

Pan, Huiyun; Li, Xiaolu; Xu, Xiaohua; Gao, Shixiang

2009-01-01

The physiological effects of 4 herbicides (butachlor, quinclorac, bensulfuron-methyl and atrazine) on 3 submerged macrophytes (Ceratophyllum demersum, Vallisneria natans and Elodea nuttallii) were tested in laboratory. The variables of the relative growth rate and the photosynthetic pigment content showed that all of the tested herbicides affected the growth of the plants obviously, even at the lowest concentration (0.0001 mg/L). Except for the C. demersum treated with quinclorac at 0.005 and 0.01 mg/L, the relative growth rates of the plants were inhibited significantly (p < 0.01). Statistical analysis of chlorophyll a (Chl-a) contents was carried out with both the t-test and one-way ANOVA to determine the difference between the treatment and control. The results showed that Chl-a contents of the plants in all treatment groups were affected by herbicides significantly, except for the C. demersum treated with bensulfuron-methyl at 0.0005 mg/L. The decrease in Chl-a content was positively correlated to the dosage of the herbicides in most treatment groups. It was suggested that herbicides in water bodies might potentially affect the growth of aquatic macrophytes. Since the Chl-a content of submerged macrophytes responded to the stress of herbicides sensitively and directly, it could be used as a biomaker in environmental monitoring or in the ecological risk assessment of herbicide contamination.

Consequences of C4 differentiation for chloroplast membrane proteomes in maize mesophyll and bundle sheath cells.

Science.gov (United States)

Majeran, Wojciech; Zybailov, Boris; Ytterberg, A Jimmy; Dunsmore, Jason; Sun, Qi; van Wijk, Klaas J

2008-09-01

Chloroplasts of maize leaves differentiate into specific bundle sheath (BS) and mesophyll (M) types to accommodate C(4) photosynthesis. Chloroplasts contain thylakoid and envelope membranes that contain the photosynthetic machineries and transporters but also proteins involved in e.g. protein homeostasis. These chloroplast membranes must be specialized within each cell type to accommodate C(4) photosynthesis and regulate metabolic fluxes and activities. This quantitative study determined the differentiated state of BS and M chloroplast thylakoid and envelope membrane proteomes and their oligomeric states using innovative gel-based and mass spectrometry-based protein quantifications. This included native gels, iTRAQ, and label-free quantification using an LTQ-Orbitrap. Subunits of Photosystems I and II, the cytochrome b(6)f, and ATP synthase complexes showed average BS/M accumulation ratios of 1.6, 0.45, 1.0, and 1.33, respectively, whereas ratios for the light-harvesting complex I and II families were 1.72 and 0.68, respectively. A 1000-kDa BS-specific NAD(P)H dehydrogenase complex with associated proteins of unknown function containing more than 15 proteins was observed; we speculate that this novel complex possibly functions in inorganic carbon concentration when carboxylation rates by ribulose-bisphosphate carboxylase/oxygenase are lower than decarboxylation rates by malic enzyme. Differential accumulation of thylakoid proteases (Egy and DegP), state transition kinases (STN7,8), and Photosystem I and II assembly factors was observed, suggesting that cell-specific photosynthetic electron transport depends on post-translational regulatory mechanisms. BS/M ratios for inner envelope transporters phosphoenolpyruvate/P(i) translocator, Dit1, Dit2, and Mex1 were determined and reflect metabolic fluxes in carbon metabolism. A wide variety of hundreds of other proteins showed differential BS/M accumulation. Mass spectral information and functional annotations are

Oxidative stress responses of submerged macrophyte Vallisneria asiatica to different concentrations of cyanobacteria

Science.gov (United States)

Kang, Caixia; Kuba, Takahiro; Hao, Aimin; Iseri, Yasushi; Li, Chunjie; Zhang, Zhenjia

2015-03-01

In a 10-day aquarium experiment, this investigation examines macrophyte restoration in eutrophic Lake Taihu, the physiological effects of different plant biomass levels and of increasing natural cyanobacterial concentrations on a submerged macrophyte, Vallisneria asiatica. Cyanobacterial stress suppressed the superoxide dismutase (SOD) activity of the plant's leaves and induced the catalase (CAT) and peroxidase (POD) activities of its roots. The soluble protein content in V. asiatica decreased with an increase in natural cyanobacterial concentrations, whereas the malonaldehyde (MDA) increased significantly at chlorophyll a (Chl a) concentrations of 222 and 262 μg/L in water. V. asiatica adapted to the stress caused by cyanobacterial concentrations by adjusting its antioxidant defense system to remove the excessive reactive oxygen species when the algal Chl a concentration was >109 μg/L. Additionally, high biomass of V. asiatica (2 222 g FW/m2) can inhibit the reproduction of cyanobacteria more significantly than low biomass (1 111 g FW/m2). High biomass of V. asiatica increased the oxidative stress in an individual plant when the initial Chl a concentration in the water reached 222 and 262 μg/L, as expressed by the increased MDA in leaves, compared with low biomass of V. asiatica. This provides a basis for controlling cyanobacterial concentrations and V. asiatica biomass for the recovery of V. asiatica in eutrophic Lake Taihu.

Chloroplast behaviour and interactions with other organelles in Arabidopsis thaliana pavement cells.

Science.gov (United States)

Barton, Kiah A; Wozny, Michael R; Mathur, Neeta; Jaipargas, Erica-Ashley; Mathur, Jaideep

2018-01-29

Chloroplasts are a characteristic feature of green plants. Mesophyll cells possess the majority of chloroplasts and it is widely believed that, with the exception of guard cells, the epidermal layer in most higher plants does not contain chloroplasts. However, recent observations on Arabidopsis thaliana have shown a population of chloroplasts in pavement cells that are smaller than mesophyll chloroplasts and have a high stroma to grana ratio. Here, using stable transgenic lines expressing fluorescent proteins targeted to the plastid stroma, plasma membrane, endoplasmic reticulum, tonoplast, nucleus, mitochondria, peroxisomes, F-actin and microtubules, we characterize the spatiotemporal relationships between the pavement cell chloroplasts (PCCs) and their subcellular environment. Observations on the PCCs suggest a source-sink relationship between the epidermal and the mesophyll layers, and experiments with the Arabidopsis mutants glabra2 ( gl2 ) and immutans ( im ), which show altered epidermal plastid development, underscored their developmental plasticity. Our findings lay down the foundation for further investigations aimed at understanding the precise role and contributions of PCCs in plant interactions with the environment. © 2018. Published by The Company of Biologists Ltd.

Phloem sap proteins from Cucurbita maxima and Ricinus communis have the capacity to traffic cell to cell through plasmodesmata.

Science.gov (United States)

Balachandran, S; Xiang, Y; Schobert, C; Thompson, G A; Lucas, W J

1997-12-09

In angiosperms, the functional enucleate sieve tube system of the phloem appears to be maintained by the surrounding companion cells. In this study, we tested the hypothesis that polypeptides present within the phloem sap traffic cell to cell from the companion cells, where they are synthesized, into the sieve tube via plasmodesmata. Coinjection of fluorescently labeled dextrans along with size-fractionated Cucurbita maxima phloem proteins, ranging in size from 10 to 200 kDa, as well as injection of individual fluorescently labeled phloem proteins, provided unambiguous evidence that these proteins have the capacity to interact with mesophyll plasmodesmata in cucurbit cotyledons to induce an increase in size exclusion limit and traffic cell to cell. Plasmodesmal size exclusion limit increased to greater than 20 kDa, but less than 40 kDa, irrespective of the size of the injected protein, indicating that partial protein unfolding may be a requirement for transport. A threshold concentration in the 20-100 nM range was required for cell-to-cell transport indicating that phloem proteins have a high affinity for the mesophyll plasmodesmal binding site(s). Parallel experiments with glutaredoxin and cystatin, phloem sap proteins from Ricinus communis, established that these proteins can also traffic through cucurbit mesophyll plasmodesmata. These results are discussed in terms of the requirements for regulated protein trafficking between companion cells and the sieve tube system. As the threshold value for plasmodesmal transport of phloem sap proteins falls within the same range as many plant hormones, the possibility is discussed that some of these proteins may act as long-distance signaling molecules.

Interaction of E. coli DNA with tobacco mesophyll protoplasts

International Nuclear Information System (INIS)

Heyn, R.F.

1975-01-01

This chapter is part of a dissertation dealing with the interaction of DNA with protoplasts. Having established the length of time during which tobacco mesophyll protoplasts do not synthesize DNA following their isolation, it is important to know the extent of DNA uptake just before the onset of DNA synthesis (and possible integration) and to find optimal conditions for this uptake. Therefore, the association of E. coli DNA with tobacco protoplasts was studied. Care should be taken with the interpretation of ''uptake'' results: adsorption phenomena play a very important role and may do so at the plasmalemma of naked protoplasts. To solve the problems involved, the use of radiation-damaged DNA was attempted. With E. coli DNA possessing a large number of thymine containing pyrimidine dimers, the loss of dimers from DNA recovered from treated protoplasts was tested in order to obtain an indication of ''real'' uptake. The results are reported

Asymmetrical effects of mesophyll conductance on fundamental photosynthetic parameters and their relationships estimated from leaf gas exchange measurements

Science.gov (United States)

Most previous analyses of leaf gas exchange measurements assumed an infinite value of mesophyll conductance (gm) and thus equaled CO2 partial pressures in the substomatal cavity and chloroplast. Yet an increasing number of studies have recognized that gm is finite and there is a drawdown of CO2 part...

Cell-specific vacuolar calcium storage mediated by CAX1 regulates apoplastic calcium concentration, gas exchange, and plant productivity in Arabidopsis.

Science.gov (United States)

Conn, Simon J; Gilliham, Matthew; Athman, Asmini; Schreiber, Andreas W; Baumann, Ute; Moller, Isabel; Cheng, Ning-Hui; Stancombe, Matthew A; Hirschi, Kendal D; Webb, Alex A R; Burton, Rachel; Kaiser, Brent N; Tyerman, Stephen D; Leigh, Roger A

2011-01-01

The physiological role and mechanism of nutrient storage within vacuoles of specific cell types is poorly understood. Transcript profiles from Arabidopsis thaliana leaf cells differing in calcium concentration ([Ca], epidermis 60 mM) were compared using a microarray screen and single-cell quantitative PCR. Three tonoplast-localized Ca(2+) transporters, CAX1 (Ca(2+)/H(+)-antiporter), ACA4, and ACA11 (Ca(2+)-ATPases), were identified as preferentially expressed in Ca-rich mesophyll. Analysis of respective loss-of-function mutants demonstrated that only a mutant that lacked expression of both CAX1 and CAX3, a gene ectopically expressed in leaves upon knockout of CAX1, had reduced mesophyll [Ca]. Reduced capacity for mesophyll Ca accumulation resulted in reduced cell wall extensibility, stomatal aperture, transpiration, CO(2) assimilation, and leaf growth rate; increased transcript abundance of other Ca(2+) transporter genes; altered expression of cell wall-modifying proteins, including members of the pectinmethylesterase, expansin, cellulose synthase, and polygalacturonase families; and higher pectin concentrations and thicker cell walls. We demonstrate that these phenotypes result from altered apoplastic free [Ca(2+)], which is threefold greater in cax1/cax3 than in wild-type plants. We establish CAX1 as a key regulator of apoplastic [Ca(2+)] through compartmentation into mesophyll vacuoles, a mechanism essential for optimal plant function and productivity.

Stomatal conductance, mesophyll conductance, and trans piration efficiency in relation to leaf anatomy in rice and wheat genotypes under drought

NARCIS (Netherlands)

Ouyang, Wenjing; Struik, Paul C.; Yin, Xinyou; Yang, Jianchang

2017-01-01

Increasing leaf transpiration efficiency (TE) may provide leads for growing rice like dryland cereals such as wheat (Triticum aestivum). To explore avenues for improving TE in rice, variations in stomatal conductance (g s) and mesophyll conductance (g m) and their anatomical determinants were

Phloem sap proteins from Cucurbita maxima and Ricinus communis have the capacity to traffic cell to cell through plasmodesmata

Science.gov (United States)

Balachandran, Suchandra; Xiang, Yu; Schobert, Christian; Thompson, Gary A.; Lucas, William J.

1997-01-01

In angiosperms, the functional enucleate sieve tube system of the phloem appears to be maintained by the surrounding companion cells. In this study, we tested the hypothesis that polypeptides present within the phloem sap traffic cell to cell from the companion cells, where they are synthesized, into the sieve tube via plasmodesmata. Coinjection of fluorescently labeled dextrans along with size-fractionated Cucurbita maxima phloem proteins, ranging in size from 10 to 200 kDa, as well as injection of individual fluorescently labeled phloem proteins, provided unambiguous evidence that these proteins have the capacity to interact with mesophyll plasmodesmata in cucurbit cotyledons to induce an increase in size exclusion limit and traffic cell to cell. Plasmodesmal size exclusion limit increased to greater than 20 kDa, but less than 40 kDa, irrespective of the size of the injected protein, indicating that partial protein unfolding may be a requirement for transport. A threshold concentration in the 20–100 nM range was required for cell-to-cell transport indicating that phloem proteins have a high affinity for the mesophyll plasmodesmal binding site(s). Parallel experiments with glutaredoxin and cystatin, phloem sap proteins from Ricinus communis, established that these proteins can also traffic through cucurbit mesophyll plasmodesmata. These results are discussed in terms of the requirements for regulated protein trafficking between companion cells and the sieve tube system. As the threshold value for plasmodesmal transport of phloem sap proteins falls within the same range as many plant hormones, the possibility is discussed that some of these proteins may act as long-distance signaling molecules. PMID:9391168

Excess Diffuse Light Absorption in Upper Mesophyll Limits CO2 Drawdown and Depresses Photosynthesis.

Science.gov (United States)

Earles, J Mason; Théroux-Rancourt, Guillaume; Gilbert, Matthew E; McElrone, Andrew J; Brodersen, Craig R

2017-06-01

In agricultural and natural systems, diffuse light can enhance plant primary productivity due to deeper penetration into and greater irradiance of the entire canopy. However, for individual sun-grown leaves from three species, photosynthesis is actually less efficient under diffuse compared with direct light. Despite its potential impact on canopy-level productivity, the mechanism for this leaf-level diffuse light photosynthetic depression effect is unknown. Here, we investigate if the spatial distribution of light absorption relative to electron transport capacity in sun- and shade-grown sunflower ( Helianthus annuus ) leaves underlies its previously observed diffuse light photosynthetic depression. Using a new one-dimensional porous medium finite element gas-exchange model parameterized with light absorption profiles, we found that weaker penetration of diffuse versus direct light into the mesophyll of sun-grown sunflower leaves led to a more heterogenous saturation of electron transport capacity and lowered its CO 2 concentration drawdown capacity in the intercellular airspace and chloroplast stroma. This decoupling of light availability from photosynthetic capacity under diffuse light is sufficient to generate an 11% decline in photosynthesis in sun-grown but not shade-grown leaves, primarily because thin shade-grown leaves similarly distribute diffuse and direct light throughout the mesophyll. Finally, we illustrate how diffuse light photosynthetic depression could overcome enhancement in canopies with low light extinction coefficients and/or leaf area, pointing toward a novel direction for future research. © 2017 American Society of Plant Biologists. All Rights Reserved.

Response of Vallisneria natans to Increasing Nitrogen Loading Depends on Sediment Nutrient Characteristics

Directory of Open Access Journals (Sweden)

Jiao Gu

2016-11-01

Full Text Available High nitrogen (N loading may contribute to recession of submerged macrophytes in shallow lakes; yet, its influences vary depending on environmental conditions. In August 2013, we conducted a 28-day factorial-designed field mesocosm experiment in Lake Taihu at the Taihu Laboratory for Lake Ecosystem Research (TLLER to examine the effects of high N loading on the growth of Vallisneria natans in systems with contrasting sediment types. We ran the experiments with two levels of nutrient loading—present-day external nutrient loading (average P: 5 μg·L−1·day−1, N: 130 μg·L−1·day−1 and P: 5 μg·L−1·day−1, and with three times higher N loading (N: 390 μg·L−1·day−1 and used sediment with two contrasting nutrient levels. V. natans growth decreased significantly with increasing N loading, the effect being dependent, however, on the nutrient status of the sediment. In low nutrient sediment, relative growth rates, leaf biomass and root biomass decreased by 11.9%, 18.2% and 23.3%, respectively, at high rather than low N loading, while the decline was larger (44.0%, 32.7% and 41.8%, respectively when using high nutrient sediment. The larger effect in the nutrient-rich sediment may reflect an observed higher shading of phytoplankton and excess nutrient accumulation in plant tissue, though potential toxic effects of the high-nutrient sediment may also have contributed. Our study confirms the occurrence of a negative effect of increasing N loading on submerged plant growth in shallow nutrient-enriched lakes and further shows that this effect is augmented when the plants grow in nutrient-rich sediment. External N control may, therefore, help to protect or restore submerged macrophytes, especially when the sediment is enriched with nutrients and organic matter.

Mesophyll conductance to CO2 transport estimated by two independent methods: effect of variable CO2 concentration and abscisic acid

Czech Academy of Sciences Publication Activity Database

Vrábl, D.; Vašková, M.; Hronková, Marie; Flexas, J.; Šantrůček, Jiří

2009-01-01

Roč. 60, č. 8 (2009), s. 2315-2323 ISSN 0022-0957 R&D Projects: GA AV ČR(CZ) IAA601410505 Institutional research plan: CEZ:AV0Z50510513 Keywords : Carbon dioxide * mesophyll conductance * Helianthus annuus Subject RIV: ED - Physiology Impact factor: 4.271, year: 2009

Opportunities for Phytoremediation and Bioindication of Arsenic Contaminated Water Using a Submerged Aquatic Plant:Vallisneria natans (lour.) Hara.

Science.gov (United States)

Chen, Guoliang; Liu, Xingmei; Brookes, Philip C; Xu, Jianming

2015-01-01

The identification of plants with high arsenic hyperaccumulating efficiency from water is required to ensure the successful application of phytoremediation technology. Five dominant submerged plant species (Vallisneria natans (Lour.) Hara., Potamageton crispus L., Myriophyllum spicatum L., Ceratophyllum demersum L. and Hydrilla verticillata (L.f.) Royle) in China were used to determine their potential to remove As from contaminated water. V. natans had the highest accumulation of As among them. The characteristics of As accumulation, transformation and the effect of phosphate on As accumulation in V. natans were then further studied. The growth of V. natans was not inhibited even when the As concentration reached 2.0 mg L(-1). After 21 d of As treatment, the bioconcentration factor (BCF) reached 1300. The As concentration in the environment and exposure time are major factors controlling the As concentration in V. natans. After being absorbed, As(V) is efficiently reduced to As(III) in plants. The synthesis of non-enzymic antioxidants may play an important role under As stress and increase As detoxication. In addition, As(V) uptake by V. natans was negatively correlated with phosphate (P) uptake when P was sufficiently supplied. As(V) is probably taken up via P transporters in V. natans.

Protoplast isolation, transient transformation of leaf mesophyll protoplasts and improved Agrobacterium-mediated leaf disc infiltration of Phaseolus vulgaris: tools for rapid gene expression analysis.

Science.gov (United States)

Nanjareddy, Kalpana; Arthikala, Manoj-Kumar; Blanco, Lourdes; Arellano, Elizabeth S; Lara, Miguel

2016-06-24

Phaseolus vulgaris is one of the most extensively studied model legumes in the world. The P. vulgaris genome sequence is available; therefore, the need for an efficient and rapid transformation system is more imperative than ever. The functional characterization of P. vulgaris genes is impeded chiefly due to the non-amenable nature of Phaseolus sp. to stable genetic transformation. Transient transformation systems are convenient and versatile alternatives for rapid gene functional characterization studies. Hence, the present work focuses on standardizing methodologies for protoplast isolation from multiple tissues and transient transformation protocols for rapid gene expression analysis in the recalcitrant grain legume P. vulgaris. Herein, we provide methodologies for the high-throughput isolation of leaf mesophyll-, flower petal-, hypocotyl-, root- and nodule-derived protoplasts from P. vulgaris. The highly efficient polyethylene glycol-mannitol magnesium (PEG-MMG)-mediated transformation of leaf mesophyll protoplasts was optimized using a GUS reporter gene. We used the P. vulgaris SNF1-related protein kinase 1 (PvSnRK1) gene as proof of concept to demonstrate rapid gene functional analysis. An RT-qPCR analysis of protoplasts that had been transformed with PvSnRK1-RNAi and PvSnRK1-OE vectors showed the significant downregulation and ectopic constitutive expression (overexpression), respectively, of the PvSnRK1 transcript. We also demonstrated an improved transient transformation approach, sonication-assisted Agrobacterium-mediated transformation (SAAT), for the leaf disc infiltration of P. vulgaris. Interestingly, this method resulted in a 90 % transformation efficiency and transformed 60-85 % of the cells in a given area of the leaf surface. The constitutive expression of YFP further confirmed the amenability of the system to gene functional characterization studies. We present simple and efficient methodologies for protoplast isolation from multiple P

Post-Transcriptional Regulation Prevents Accumulation of Glutathione Reductase Protein and Activity in the Bundle Sheath Cells of Maize1

Science.gov (United States)

Pastori, Gabriela M.; Mullineaux, Philip M.; Foyer, Christine H.

2000-01-01

Glutathione reductase (GR; EC 1.6.4.2) activity was assayed in bundle sheath and mesophyll cells of maize (Zea mays L. var H99) from plants grown at 20°C, 18°C, and 15°C. The purity of each fraction was determined by measuring the associated activity of the compartment-specific marker enzymes, Rubisco and phosphoenolpyruvate carboxylase, respectively. GR activity and the abundance of GR protein and mRNA increased in plants grown at 15°C and 18°C compared with those grown at 20°C. In all cases GR activity was found only in mesophyll fractions of the leaves, with no GR activity being detectable in bundle sheath extracts. Immunogold labeling with GR-specific antibodies showed that the GR protein was exclusively localized in the mesophyll cells of leaves at all growth temperatures, whereas GR transcripts (as determined by in situ hybridization techniques) were observed in both cell types. These results indicate that post-transcriptional regulation prevents GR accumulation in the bundle sheath cells of maize leaves. The resulting limitation on the capacity for regeneration of reduced glutathione in this compartment may contribute to the extreme chilling sensitivity of maize leaves. PMID:10712529

Local differentiation of cell wall matrix polysaccharides in sinuous pavement cells: its possible involvement in the flexibility of cell shape.

Science.gov (United States)

Sotiriou, P; Giannoutsou, E; Panteris, E; Galatis, B; Apostolakos, P

2018-03-01

The distribution of homogalacturonans (HGAs) displaying different degrees of esterification as well as of callose was examined in cell walls of mature pavement cells in two angiosperm and two fern species. We investigated whether local cell wall matrix differentiation may enable pavement cells to respond to mechanical tension forces by transiently altering their shape. HGA epitopes, identified with 2F4, JIM5 and JIM7 antibodies, and callose were immunolocalised in hand-made or semithin leaf sections. Callose was also stained with aniline blue. The structure of pavement cells was studied with light and transmission electron microscopy (TEM). In all species examined, pavement cells displayed wavy anticlinal cell walls, but the waviness pattern differed between angiosperms and ferns. The angiosperm pavement cells were tightly interconnected throughout their whole depth, while in ferns they were interconnected only close to the external periclinal cell wall and intercellular spaces were developed between them close to the mesophyll. Although the HGA epitopes examined were located along the whole cell wall surface, the 2F4- and JIM5- epitopes were especially localised at cell lobe tips. In fern pavement cells, the contact sites were impregnated with callose and JIM5-HGA epitopes. When tension forces were applied on leaf regions, the pavement cells elongated along the stretching axis, due to a decrease in waviness of anticlinal cell walls. After removal of tension forces, the original cell shape was resumed. The presented data support that HGA epitopes make the anticlinal pavement cell walls flexible, in order to reversibly alter their shape. Furthermore, callose seems to offer stability to cell contacts between pavement cells, as already suggested in photosynthetic mesophyll cells. © 2017 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.

The effects of cadmium pulse dosing on physiological traits and growth of the submerged macrophyte Vallisneria spinulosa and phytoplankton biomass: a mesocosm study.

Science.gov (United States)

Liu, Hui; Cao, Yu; Li, Wei; Zhang, Zhao; Jeppesen, Erik; Wang, Wei

2017-06-01

Pulse inputs of heavy metals are expected to increase with a higher frequency of extreme climate events (heavy rain), leading to stronger erosion of contaminated and fertilized farmland soils to freshwaters, with potentially adverse effects on lake ecosystems. We conducted a 5-month mesocosm study to elucidate the responses of the submerged macrophyte Vallisneria spinulosa and phytoplankton to four different doses of cadmium (Cd): 0 (control), 0.05, 0.5, and 5 g m -2 (CK, I, II, and III, respectively) under mesotrophic conditions. We found that total phosphorus concentrations were larger in the three Cd pulse treatments, whereas total nitrogen concentrations did not differ among the four treatments. The contents of chlorophyll a and soluble sugar in macrophyte leaves decreased in III, and total biomass, ramet number, plant height, and total stolon length of macrophytes were lower in both II and III. In contrast, abundances of the three main phytoplankton taxa-Cyanophyta, Chlorophyta, and Bacillariophyta-did not differ among treatments. Total phytoplankton biomass was, however, marginally lower in CK than in the Cd treatments. We conclude that exposure to strong Cd pulses led to significantly reduced growth of macrophytes, while no obvious effect appeared for phytoplankton.

Patterns of indole alkaloids synthesis in response to heat shock, 5-azacytidine and Na-butyrate treatment of cultured catharanthus roseus mesophyll protoplasts

International Nuclear Information System (INIS)

Saleem, M.; Cutler, A.J.

1986-01-01

Alkaloids of C. roseus are in high demand for therapeutic and other reasons. Cultured Catharanthus cells can produce limited quantities of these alkaloids. The authors have found that cultured mesophyll protoplasts in the presence of 14 C-Tryptamine are capable of synthesizing alkaloids. The pattern of alkaloids synthesis changes when protoplasts are subjected to a heat shock at 37 0 C. The heat shocked protoplasts incorporated 33% more 14 C-Tryptamine and produced 3 new types of alkaloids. Treatment of protoplasts with 5-azacytidine, a DNA hypomethylating agent and Na-butyrate which induces hyperacetylation of histones produced qualitative and quantitative changes in the alkaloid pattern. Four new alkaloids following the above treatments were detected by TLC and HPLC of the extracts. It is suggested that the alkaloid pattern of the cultured protoplasts can be altered by treatment with compounds known as regulators of gene expression. Work is in progress to isolate and identify these new alkaloids

Variability in mesophyll conductance between barley genotypes, and effects on transpiration efficiency and carbon isotope discrimination.

Science.gov (United States)

Barbour, Margaret M; Warren, Charles R; Farquhar, Graham D; Forrester, Guy; Brown, Hamish

2010-07-01

Leaf internal, or mesophyll, conductance to CO(2) (g(m)) is a significant and variable limitation of photosynthesis that also affects leaf transpiration efficiency (TE). Genotypic variation in g(m) and the effect of g(m) on TE were assessed in six barley genotypes (four Hordeum vulgare and two H. bulbosum). Significant variation in g(m) was found between genotypes, and was correlated with photosynthetic rate. The genotype with the highest g(m) also had the highest TE and the lowest carbon isotope discrimination as recorded in leaf tissue (Delta(p)). These results suggest g(m) has unexplored potential to provide TE improvement within crop breeding programmes.

Photosynthesis in Flaveria brownii, a C(4)-Like Species: Leaf Anatomy, Characteristics of CO(2) Exchange, Compartmentation of Photosynthetic Enzymes, and Metabolism of CO(2).

Science.gov (United States)

Cheng, S H; Moore, B D; Edwards, G E; Ku, M S

1988-08-01

Light microscopic examination of leaf cross-sections showed that Flaveria brownii A. M. Powell exhibits Kranz anatomy, in which distinct, chloroplast-containing bundle sheath cells are surrounded by two types of mesophyll cells. Smaller mesophyll cells containing many chloroplasts are arranged around the bundle sheath cells. Larger, spongy mesophyll cells, having fewer chloroplasts, are located between the smaller mesophyll cells and the epidermis. F. brownii has very low CO(2) compensation points at different O(2) levels, which is typical of C(4) plants, yet it does show about 4% inhibition of net photosynthesis by 21% O(2) at 30 degrees C. Protoplasts of the three photosynthetic leaf cell types were isolated according to relative differences in their buoyant densities. On a chlorophyll basis, the activities of phosphoenolpyruvate carboxylase and pyruvate, Pi dikinase (carboxylation phase of C(4) pathway) were highest in the larger mesophyll protoplasts, intermediate in the smaller mesophyll protoplasts, and lowest, but still present, in the bundle sheath protoplasts. In contrast, activities of ribulose 1,5-bisphosphate carboxylase, other C(3) cycle enzymes, and NADP-malic enzyme showed a reverse gradation, although there were significant activities of these enzymes in mesophyll cells. As indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the banding pattern of certain polypeptides of the total soluble proteins from the three cell types also supported the distribution pattern obtained by activity assays of these enzymes. Analysis of initial (14)C products in whole leaves and extrapolation of pulse-labeling curves to zero time indicated that about 80% of the CO(2) is fixed into C(4) acids (malate and aspartate), whereas about 20% of the CO(2) directly enters the C(3) cycle. This is consistent with the high activity of enzymes for CO(2) fixation by the C(4) pathway and the substantial activity of enzymes of the C(3) cycle in the mesophyll cells

Carbon dioxide diffusion across stomata and mesophyll and photo-biochemical processes as affected by growth CO2 and phosphorus nutrition in cotton.

Science.gov (United States)

Singh, Shardendu K; Badgujar, Girish; Reddy, Vangimalla R; Fleisher, David H; Bunce, James A

2013-06-15

Nutrients such as phosphorus may exert a major control over plant response to rising atmospheric carbon dioxide concentration (CO2), which is projected to double by the end of the 21st century. Elevated CO2 may overcome the diffusional limitations to photosynthesis posed by stomata and mesophyll and alter the photo-biochemical limitations resulting from phosphorus deficiency. To evaluate these ideas, cotton (Gossypium hirsutum) was grown in controlled environment growth chambers with three levels of phosphate (Pi) supply (0.2, 0.05 and 0.01mM) and two levels of CO2 concentration (ambient 400 and elevated 800μmolmol(-1)) under optimum temperature and irrigation. Phosphate deficiency drastically inhibited photosynthetic characteristics and decreased cotton growth for both CO2 treatments. Under Pi stress, an apparent limitation to the photosynthetic potential was evident by CO2 diffusion through stomata and mesophyll, impairment of photosystem functioning and inhibition of biochemical process including the carboxylation efficiency of ribulose-1,5-bisphosphate carboxylase/oxyganase and the rate of ribulose-1,5-bisphosphate regeneration. The diffusional limitation posed by mesophyll was up to 58% greater than the limitation due to stomatal conductance (gs) under Pi stress. As expected, elevated CO2 reduced these diffusional limitations to photosynthesis across Pi levels; however, it failed to reduce the photo-biochemical limitations to photosynthesis in phosphorus deficient plants. Acclimation/down regulation of photosynthetic capacity was evident under elevated CO2 across Pi treatments. Despite a decrease in phosphorus, nitrogen and chlorophyll concentrations in leaf tissue and reduced stomatal conductance at elevated CO2, the rate of photosynthesis per unit leaf area when measured at the growth CO2 concentration tended to be higher for all except the lowest Pi treatment. Nevertheless, plant biomass increased at elevated CO2 across Pi nutrition with taller plants

Proton extrusion is an essential signalling component in the HR of epidermal single cells in the barley-powdery mildew interaction

DEFF Research Database (Denmark)

Zhou, F.S.; Andersen, C.H.; Burhenne, K.

2000-01-01

We propose a model for activation of the epidermal cell hypersensitive response (HR) in the barley/powdery mildew interaction. The model suggests that the plasma membrane proton pump (H+-ATPase) of epidermal cells is activated following penetration by an avirulent powdery mildew fungus...... in the incompatible interaction; (4) race-specific proton extrusion is observed underneath epidermal tissue detached from leaves inoculated 15 h earlier; and (5) treatment of leaves with fusicoccin, an activator of the plasma membrane H+-ATPase, increases the number of HR-cells in the compatible interaction........ This will cause an acidification of the apoplast towards the mesophyll cells, thereby activating generation of H2O2 from the mesophyll, which subsequently triggers the epidermal cell to undergo HR. The model is supported by the following data: (1) the earliest HR-related H2O2 is found in the attachment zones...

Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

Science.gov (United States)

Li, Jian-Feng; Bush, Jenifer; Xiong, Yan; Li, Lei; McCormack, Matthew

2011-01-01

Protein-protein interactions (PPIs) constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC) as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs) and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

Directory of Open Access Journals (Sweden)

Jian-Feng Li

Full Text Available Protein-protein interactions (PPIs constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

Mesophyll conductance in Zea mays responds transiently to CO2 availability: implications for transpiration efficiency in C4 crops.

Science.gov (United States)

Kolbe, Allison R; Cousins, Asaph B

2018-03-01

Mesophyll conductance (g m ) describes the movement of CO 2 from the intercellular air spaces below the stomata to the site of initial carboxylation in the mesophyll. In contrast with C 3 -g m , little is currently known about the intraspecific variation in C 4 -g m or its responsiveness to environmental stimuli. To address these questions, g m was measured on five maize (Zea mays) lines in response to CO 2 , employing three different estimates of g m . Each of the methods indicated a significant response of g m to CO 2 . Estimates of g m were similar between methods at ambient and higher CO 2 , but diverged significantly at low partial pressures of CO 2 . These differences are probably driven by incomplete chemical and isotopic equilibrium between CO 2 and bicarbonate under these conditions. Carbonic anhydrase and phosphoenolpyruvate carboxylase in vitro activity varied significantly despite similar values of g m and leaf anatomical traits. These results provide strong support for a CO 2 response of g m in Z. mays, and indicate that g m in maize is probably driven by anatomical constraints rather than by biochemical limitations. The CO 2 response of g m indicates a potential role for facilitated diffusion in C 4 -g m . These results also suggest that water-use efficiency could be enhanced in C 4 species by targeting g m . © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

Using tunable diode laser spectroscopy to measure carbon isotope discrimination and mesophyll conductance to CO₂ diffusion dynamically at different CO₂ concentrations.

Science.gov (United States)

Tazoe, Youshi; VON Caemmerer, Susanne; Estavillo, Gonzalo M; Evans, John R

2011-04-01

In C₃ leaves, the mesophyll conductance to CO₂ diffusion, g(m) , determines the drawdown in CO₂ concentration from intercellular airspace to the chloroplast stroma. Both g(m) and stomatal conductance limit photosynthetic rate and vary in response to the environment. We investigated the response of g(m) to changes in CO₂ in two Arabidopsis genotypes (including a mutant with open stomata, ost1), tobacco and wheat. We combined measurements of gas exchange with carbon isotope discrimination using tunable diode laser absorption spectroscopy with a CO₂ calibration system specially designed for a range of CO₂ and O₂ concentrations. CO₂ was initially increased from 200 to 1000 ppm and then decreased stepwise to 200 ppm and increased stepwise back to 1000 ppm, or the sequence was reversed. In 2% O₂ a step increase from 200 to 1000 ppm significantly decreased g(m) by 26-40% in all three species, whereas following a step decrease from 1000 to 200 ppm, the 26-38% increase in g(m) was not statistically significant. The response of g(m) to CO₂ was less in 21% O₂. Comparing wild type against the ost1 revealed that mesophyll and stomatal conductance varied independently in response to CO₂. We discuss the effects of isotope fractionation factors on estimating g(m) . © 2011 Blackwell Publishing Ltd.

Photosynthetic response of an alpine plant, Rhododendron delavayi Franch, to water stress and recovery: the role of mesophyll conductance

Directory of Open Access Journals (Sweden)

Yanfei eCai

2015-12-01

Full Text Available Rhododendron delavayi Franch is an evergreen shrub or small tree with large scarlet flowers that makes it highly attractive as an ornamental species. The species is native to southwest China and southeast Asia, especially the Himalayan region, showing good adaptability and tolerance to drought. To understand the water stress coping mechanisms of R. delavayi, we analysed the plant’s photosynthetic performance during water stress and recovery. In particular, we looked at the regulation of stomatal (gs and mesophyll conductance (gm, and maximum rate of carboxylation (Vcmax. After four days of water stress treatment, the net CO2 assimilation rate (AN declined slightly while gs and gm were not affected and stomatal limitation (SL was therefore negligible. At this stage mesophyll conductance limitation (MCL and biochemical limitation (BL constituted the main limitation factors. After eight days of water stress treatment, AN, gs and gm had decreased notably. At this stage SL increased markedly and MCL even more so, while BL remained relatively constant. After re-watering, the recovery of AN, gs and gm was rapid, although remaining below the levels of the control plants, while Vcmax fully regained control levels after three days of re-watering. MCL remained the main limitation factor irrespective of the degree of photosynthetic recovery. In conclusion, in our experiment MCL was the main photosynthetic limitation factor of R. delavayi under water stress and during the recovery phase, with the regulation of gm probably being the result of interactions between the environment and leaf anatomical features.

Changes in photosynthesis, mesophyll conductance to CO2, and isoprenoid emissions in Populus nigra plants exposed to excess nickel

International Nuclear Information System (INIS)

Velikova, Violeta; Tsonev, Tsonko; Loreto, Francesco; Centritto, Mauro

2011-01-01

Poplar (Populus nigra) plants were grown hydroponically with 30 and 200 μM Ni (Ni 30 and Ni 200 ). Photosynthesis limitations and isoprenoid emissions were investigated in two leaf types (mature and developing). Ni stress significantly decreased photosynthesis, and this effect depended on the leaf Ni content, which was lower in mature than in developing leaves. The main limitations to photosynthesis were attributed to mesophyll conductance and metabolism impairment. In Ni-stressed developing leaves, isoprene emission was significantly stimulated. We attribute such stimulation to the lower chloroplastic [CO 2 ] than in control leaves. However chloroplastic [CO 2 ] did not control isoprene emission in mature leaves. Ni stress induced the emission of cis-β-ocimene in mature leaves, and of linalool in both leaf types. Induced biosynthesis and emission of isoprenoids reveal the onset of antioxidant processes that may also contribute to reduce Ni stress, especially in mature poplar leaves. - Graphical abstract: Visible damage caused by Ni treatment. 1 - Ni 0 (control plants); 2 - Ni 200 ; M = mature and D = developing Populus nigra leaves. Display Omitted Highlights: → We study the effect of Ni pollution on photosynthesis and isoprenoid emissions. → Ni stress significantly decreases photosynthesis. The main limitations are attributed to mesophyll conductance and metabolism impairment. → Constitutive isoprene emission was significantly stimulated in Ni-stressed leaves. Exposure to enhanced Ni concentration induces cis-beta-ocimene and linalool emissions. - The study reveals consequences of Ni stress on plant physiology, namely increasing diffusional limitation to photosynthesis and isoprenoid emissions.

Establishing in vitro Zinnia elegans cell suspension culture with high tracheary elements differentiation

NARCIS (Netherlands)

Twumasi, P.; Schel, J.H.N.; Ieperen, van W.; Woltering, E.J.; Emons, A.M.C.

2009-01-01

The Zinnia elegans mesophyll cell culture is a useful system for xylogenesis studies. The system is associated with highly synchronous tracheary element (TE) differentiation, making it more suitable for molecular studies requiring larger amounts of molecular isolates, such as mRNA and proteins and

The Arabidopsis arc5 and arc6 mutations differentially affect plastid morphology in pavement and guard cells in the leaf epidermis.

Science.gov (United States)

Fujiwara, Makoto T; Yasuzawa, Mana; Kojo, Kei H; Niwa, Yasuo; Abe, Tomoko; Yoshida, Shigeo; Nakano, Takeshi; Itoh, Ryuuichi D

2018-01-01

Chloroplasts, or photosynthetic plastids, multiply by binary fission, forming a homogeneous population in plant cells. In Arabidopsis thaliana, the division apparatus (or division ring) of mesophyll chloroplasts includes an inner envelope transmembrane protein ARC6, a cytoplasmic dynamin-related protein ARC5 (DRP5B), and members of the FtsZ1 and FtsZ2 families of proteins, which co-assemble in the stromal mid-plastid division ring (FtsZ ring). FtsZ ring placement is controlled by several proteins, including a stromal factor MinE (AtMinE1). During leaf mesophyll development, ARC6 and AtMinE1 are necessary for FtsZ ring formation and thus plastid division initiation, while ARC5 is essential for a later stage of plastid division. Here, we examined plastid morphology in leaf epidermal pavement cells (PCs) and stomatal guard cells (GCs) in the arc5 and arc6 mutants using stroma-targeted fluorescent proteins. The arc5 PC plastids were generally a bit larger than those of the wild type, but most had normal shapes and were division-competent, unlike mutant mesophyll chloroplasts. The arc6 PC plastids were heterogeneous in size and shape, including the formation of giant and mini-plastids, plastids with highly developed stromules, and grape-like plastid clusters, which varied on a cell-by-cell basis. Moreover, unique plastid phenotypes for stomatal GCs were observed in both mutants. The arc5 GCs rarely lacked chlorophyll-bearing plastids (chloroplasts), while they accumulated minute chlorophyll-less plastids, whereas most GCs developed wild type-like chloroplasts. The arc6 GCs produced large chloroplasts and/or chlorophyll-less plastids, as previously observed, but unexpectedly, their chloroplasts/plastids exhibited marked morphological variations. We quantitatively analyzed plastid morphology and partitioning in paired GCs from wild-type, arc5, arc6, and atminE1 plants. Collectively, our results support the notion that ARC5 is dispensable in the process of equal division

Changes in photosynthesis, mesophyll conductance to CO{sub 2}, and isoprenoid emissions in Populus nigra plants exposed to excess nickel

Energy Technology Data Exchange (ETDEWEB)

Velikova, Violeta, E-mail: violet@obzor.bio21.bas.bg [Bulgarian Academy of Sciences, Acad. M. Popov Institute of Plant Physiology, Acad. G. Bonchev, Bl. 21, 1113 Sofia (Bulgaria); Tsonev, Tsonko [Bulgarian Academy of Sciences, Acad. M. Popov Institute of Plant Physiology, Acad. G. Bonchev, Bl. 21, 1113 Sofia (Bulgaria); Loreto, Francesco [Consiglio Nazionale delle Ricerche, Istituto per la Protezione delle Piante, Via Madonna del Piano 10, 50019 Sesto Fiorentino, Firenze (Italy); Centritto, Mauro [Consiglio Nazionale delle Ricerche, Istituto di Biologia Agroambientale e Forestale, 00015 Monterotondo Scalo (RM) (Italy)

2011-05-15

Poplar (Populus nigra) plants were grown hydroponically with 30 and 200 {mu}M Ni (Ni{sub 30} and Ni{sub 200}). Photosynthesis limitations and isoprenoid emissions were investigated in two leaf types (mature and developing). Ni stress significantly decreased photosynthesis, and this effect depended on the leaf Ni content, which was lower in mature than in developing leaves. The main limitations to photosynthesis were attributed to mesophyll conductance and metabolism impairment. In Ni-stressed developing leaves, isoprene emission was significantly stimulated. We attribute such stimulation to the lower chloroplastic [CO{sub 2}] than in control leaves. However chloroplastic [CO{sub 2}] did not control isoprene emission in mature leaves. Ni stress induced the emission of cis-{beta}-ocimene in mature leaves, and of linalool in both leaf types. Induced biosynthesis and emission of isoprenoids reveal the onset of antioxidant processes that may also contribute to reduce Ni stress, especially in mature poplar leaves. - Graphical abstract: Visible damage caused by Ni treatment. 1 - Ni{sub 0} (control plants); 2 - Ni{sub 200}; M = mature and D = developing Populus nigra leaves. Display Omitted Highlights: > We study the effect of Ni pollution on photosynthesis and isoprenoid emissions. > Ni stress significantly decreases photosynthesis. The main limitations are attributed to mesophyll conductance and metabolism impairment. > Constitutive isoprene emission was significantly stimulated in Ni-stressed leaves. Exposure to enhanced Ni concentration induces cis-beta-ocimene and linalool emissions. - The study reveals consequences of Ni stress on plant physiology, namely increasing diffusional limitation to photosynthesis and isoprenoid emissions.

The physiological responses of Vallisneria natans to epiphytic algae with the increase of N and P concentrations in water bodies.

Science.gov (United States)

Song, Yu-Zhi; Wang, Jin-Qi; Gao, Yong-Xia; Xie, Xue-Jian

2015-06-01

To reveal the mechanism of submerged plants decline in progressively eutrophicated freshwaters, physiological responses of Vallisneria natans to epiphytic algae were studied in simulation lab by measuring plant physiological indexes of chlorophyll content, malondialdehyde (MDA) content, and superoxide dismutase (SOD) activity based on a 2 × 4 factorial design with two epiphytic conditions (with epiphytic algae and without) and four levels of N and P concentrations in water (N-P[mg.L(-1)]: 0.5, 0.05; 2.5, 0.25; 4.5, 0.45; 12.5, 1.25). Compared with control (non-presence of epiphytic algae), chlorophyll contents of V. natans were significantly decreased (p algae under any concentrations of N and P in water bodies. While the presence of epiphytic algae induced peroxidation of membrane lipids, MDA contents of V. natans had significantly increased (p algae in the treatments of T2 and T3 in the whole culture process by comparing with control, sometimes reaching an extremely significant level (p algae (p algae on chlorophyll content and SOD activity in the leaves of V. natans were increased at first and then decreased with the concentrations of N and P in water, and MDA content became higher with the increase of N and P. concentrations. Repeated measurement data testing showed that the effects of epiphytic algae on the chlorophyll content and MDA content and SOD activity were significant, respectively (p algae were combining with effects of concentrations of N and P (p algae directly produced adverse effects on physiology of V. natans and epiphytic algal biomass were positively correlated with nutrient available in the water column.

LAM-1 and FAT Genes Control Development of the Leaf Blade in Nicotiana sylvestris.

Science.gov (United States)

McHale, NA

1993-01-01

Leaf primordia of the lam-1 mutant of Nicotiana sylvestris grow normally in length but remain bladeless throughout development. The blade initiation site is established at the normal time and position in lam-1 primordia. Anticlinal divisions proceed normally in the outer L1 and L2 layers, but the inner L3 cells fail to establish the periclinal divisions that normally generate the middle mesophyll core. The lam-1 mutation also blocks formation of blade mesophyll from distal L2 cells. This suggests that LAM-1 controls a common step in initiation of blade tissue from the L2 and L3 lineage of the primordium. Another recessive mutation (fat) was isolated in N. sylvestris that induces abnormal periclinal divisions in the mesophyll during blade initiation and expansion. This generates a blade approximately twice its normal thickness by doubling the number of mesophyll cell layers from four to approximately eight. Presumably, the fat mutation defines a negative regulator involved in repression of periclinal divisions in the blade. The lam-1 fat double mutant shows radial proliferation of mesophyll cells at the blade initiation site. This produces a highly disorganized, club-shaped blade that appears to represent an additive effect of the lam-1 and fat mutations on blade founder cells. PMID:12271096

Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leaves. I. Growth can occur without photosynthesis

Science.gov (United States)

Van Volkenburgh, E.; Cleland, R. E.

1990-01-01

Cell expansion in dicotyledonous leaves is strongly stimulated by bright white light (WL), at least in part as a result of light-induced acidification of the cell walls. It has been proposed that photosynthetic reactions are required for light-stimulated transport processes across plasma membranes of leaf cells, including proton excretion. The involvement of photosynthesis in growth and wall acidification of primary leaves of bean has been tested by inhibiting photosynthesis in two ways: by reducing chlorophyll content of intact plants with tentoxin (TX) and by treating leaf discs with 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Exposure to bright WL stimulated growth of intact leaves of TX-treated plants. Discs excised from green as well as from TX-or DCMU-treated leaves also responded by growing faster in WL, as long as exogenous sucrose was supplied to the photosynthetically inhibited tissues. The WL caused acidification of the epidermal surface of intact TX-leaves, but acidification of the incubation medium by mesophyll cells only occurred when photosynthesis was not inhibited. It is concluded that light-stimulated cell enlargement of bean leaves, and the necessary acidification of epidermal cell walls, are mediated by a pigment other than chlorophyll. Light-induced proton excretion by mesophyll cells, on the other hand, may require both a photosynthetic product (or exogenous sugars) and a non-photosynthetic light effect.

Longevity of guard cell chloroplasts in falling leaves: implication for stomatal function and cellular aging.

Science.gov (United States)

Zeiger, E; Schwartz, A

1982-11-12

Guard cell chloroplasts in senescing leaves from 12 species of perennial trees and three species of annual plants survived considerably longer than their mesophyll counterparts. In Ginkgo biloba, stomata from yellow leaves opened during the day and closed at night; guard cell chloroplasts from these leaves showed fluorescence transients associated with electron transport and photophosphorylation. These findings indicate that guard cell chloroplasts are highly conserved throughout the life-span of the leaf and that leaves retain stomatal control during senescence.

Phloem Loading in Two Scrophulariaceae Species. What Can Drive Symplastic Flow via Plasmodesmata?1

Science.gov (United States)

Voitsekhovskaja, Olga V.; Koroleva, Olga A.; Batashev, Denis R.; Knop, Christian; Tomos, A. Deri; Gamalei, Yuri V.; Heldt, Hans-Walter; Lohaus, Gertrud

2006-01-01

To determine the driving forces for symplastic sugar flux between mesophyll and phloem, gradients of sugar concentrations and osmotic pressure were studied in leaf tissues of two Scrophulariaceae species, Alonsoa meridionalis and Asarina barclaiana. A. meridionalis has a typical symplastic configuration of minor-vein phloem, i.e. intermediary companion cells with highly developed plasmodesmal connections to bundle-sheath cells. In A. barclaiana, two types of companion cells, modified intermediary cells and transfer cells, were found in minor-vein phloem, giving this species the potential to have a complex phloem-loading mode. We identified all phloem-transported carbohydrates in both species and analyzed the levels of carbohydrates in chloroplasts, vacuoles, and cytoplasm of mesophyll cells by nonaqueous fractionation. Osmotic pressure was measured in single epidermal and mesophyll cells and in whole leaves and compared with calculated values for phloem sap. In A. meridionalis, a 2-fold concentration gradient for sucrose between mesophyll and phloem was found. In A. barclaiana, the major transported carbohydrates, sucrose and antirrhinoside, were present in the phloem in 22- and 6-fold higher concentrations, respectively, than in the cytoplasm of mesophyll cells. The data show that diffusion of sugars along their concentration gradients is unlikely to be the major mechanism for symplastic phloem loading if this were to occur in these species. We conclude that in both A. meridionalis and A. barclaiana, apoplastic phloem loading is an indispensable mechanism and that symplastic entrance of solutes into the phloem may occur by mass flow. The conditions favoring symplastic mass flow into the phloem are discussed. PMID:16377750

Phloem loading in two Scrophulariaceae species. What can drive symplastic flow via plasmodesmata?

Science.gov (United States)

Voitsekhovskaja, Olga V; Koroleva, Olga A; Batashev, Denis R; Knop, Christian; Tomos, A Deri; Gamalei, Yuri V; Heldt, Hans-Walter; Lohaus, Gertrud

2006-01-01

To determine the driving forces for symplastic sugar flux between mesophyll and phloem, gradients of sugar concentrations and osmotic pressure were studied in leaf tissues of two Scrophulariaceae species, Alonsoa meridionalis and Asarina barclaiana. A. meridionalis has a typical symplastic configuration of minor-vein phloem, i.e. intermediary companion cells with highly developed plasmodesmal connections to bundle-sheath cells. In A. barclaiana, two types of companion cells, modified intermediary cells and transfer cells, were found in minor-vein phloem, giving this species the potential to have a complex phloem-loading mode. We identified all phloem-transported carbohydrates in both species and analyzed the levels of carbohydrates in chloroplasts, vacuoles, and cytoplasm of mesophyll cells by nonaqueous fractionation. Osmotic pressure was measured in single epidermal and mesophyll cells and in whole leaves and compared with calculated values for phloem sap. In A. meridionalis, a 2-fold concentration gradient for sucrose between mesophyll and phloem was found. In A. barclaiana, the major transported carbohydrates, sucrose and antirrhinoside, were present in the phloem in 22- and 6-fold higher concentrations, respectively, than in the cytoplasm of mesophyll cells. The data show that diffusion of sugars along their concentration gradients is unlikely to be the major mechanism for symplastic phloem loading if this were to occur in these species. We conclude that in both A. meridionalis and A. barclaiana, apoplastic phloem loading is an indispensable mechanism and that symplastic entrance of solutes into the phloem may occur by mass flow. The conditions favoring symplastic mass flow into the phloem are discussed.

Analysis of plastid number, size, and distribution in Arabidopsis plants by light and fluorescence microscopy.

Science.gov (United States)

Pyke, Kevin

2011-01-01

Methods are described which allow one to observe chloroplasts in mesophyll cells from leaves of Arabidopsis, determine their number per cell, measure their area, and determine a value for chloroplast coverage inside mesophyll cells. Non-green plastids can also be imaged either by using staining, or by exploiting fluorescent proteins targeted to the plastid in non-green parts of the plant, such as the roots, in transgenic Arabidopsis.

Anatomical changes on coffee leaves infected by Pseudomonas syringae pv. garcae

Directory of Open Access Journals (Sweden)

Lucas Mateus Rivero Rodrigues

2015-12-01

Full Text Available ABSTRACTAlthough poorly studied, the bacterial halo blight is an important disease in the major coffee-producing states of Brazil. External damage and anatomical changes on leaves were measured in seedlings of Coffea arabica cv. Mundo Novo, susceptible to Pseudomonas syringae pv. garcae, by using histological sections obtained at 10 and 20 days after inoculation (DAI. The changes on the epidermis were smaller than the lesions measured in the mesophyll, irrespective of the evaluated colonization period, showing that the internal damage caused by the bacterium represent twice the damage observed externally. From the inoculation site, lysis occurred on the epidermal cells and on the palisade and spongy parenchyma cells, with strong staining of their cellular contents, as well as abnormal intercellular spaces in the palisade parenchyma, hypertrophy and hyperplasia of mesophyll cells and partial destruction of chloroplasts. Additionally, this study revealed the presence of inclusion bodies in epidermal and mesophyll cells. Bacterial masses were found in the apoplast between and within mesophyll cells. Bacteria were also observed in the bundle sheath and vascular bundles and were more pronounced at 20 DAI, not only near the inoculation site but also in distant areas, suggesting displacement through the vascular system. These results can be useful to understand this plant-pathogen interaction.

In Vivo Quantification of Cell Coupling in Plants with Different Phloem-Loading Strategies[W][OA

Science.gov (United States)

Liesche, Johannes; Schulz, Alexander

2012-01-01

Uptake of photoassimilates into the leaf phloem is the key step in carbon partitioning and phloem transport. Symplasmic and apoplasmic loading strategies have been defined in different plant taxa based on the abundance of plasmodesmata between mesophyll and phloem. For apoplasmic loading to occur, an absence of plasmodesmata is a sufficient but not a necessary criterion, as passage of molecules through plasmodesmata might well be blocked or restricted. Here, we present a noninvasive, whole-plant approach to test symplasmic coupling and quantify the intercellular flux of small molecules using photoactivation microscopy. Quantification of coupling between all cells along the prephloem pathways of the apoplasmic loader Vicia faba and Nicotiana tabacum showed, to our knowledge for the first time in vivo, that small solutes like sucrose can diffuse through plasmodesmata up to the phloem sieve element companion cell complex (SECCC). As expected, the SECCC was found to be symplasmically isolated for small solutes. In contrast, the prephloem pathway of the symplasmic loader Cucurbita maxima was found to be well coupled with the SECCC. Phloem loading in gymnosperms is not well understood, due to a profoundly different leaf anatomy and a scarcity of molecular data compared with angiosperms. A cell-coupling analysis for Pinus sylvestris showed high symplasmic coupling along the entire prephloem pathway, comprising at least seven cell border interfaces between mesophyll and sieve elements. Cell coupling together with measurements of leaf sap osmolality indicate a passive symplasmic loading type. Similarities and differences of this loading type with that of angiosperm trees are discussed. PMID:22422939

Microarray Expression Analyses of Arabidopsis Guard Cells and Isolation of a Recessive Abscisic Acid Hypersensitive Protein Phosphatase 2C MutantW⃞

Science.gov (United States)

Leonhardt, Nathalie; Kwak, June M.; Robert, Nadia; Waner, David; Leonhardt, Guillaume; Schroeder, Julian I.

2004-01-01

Oligomer-based DNA Affymetrix GeneChips representing about one-third of Arabidopsis (Arabidopsis thaliana) genes were used to profile global gene expression in a single cell type, guard cells, identifying 1309 guard cell–expressed genes. Highly pure preparations of guard cells and mesophyll cells were isolated in the presence of transcription inhibitors that prevented induction of stress-inducible genes during cell isolation procedures. Guard cell expression profiles were compared with those of mesophyll cells, resulting in identification of 64 transcripts expressed preferentially in guard cells. Many large gene families and gene duplications are known to exist in the Arabidopsis genome, giving rise to redundancies that greatly hamper conventional genetic and functional genomic analyses. The presented genomic scale analysis identifies redundant expression of specific isoforms belonging to large gene families at the single cell level, which provides a powerful tool for functional genomic characterization of the many signaling pathways that function in guard cells. Reverse transcription–PCR of 29 genes confirmed the reliability of GeneChip results. Statistical analyses of promoter regions of abscisic acid (ABA)–regulated genes reveal an overrepresented ABA responsive motif, which is the known ABA response element. Interestingly, expression profiling reveals ABA modulation of many known guard cell ABA signaling components at the transcript level. We further identified a highly ABA-induced protein phosphatase 2C transcript, AtP2C-HA, in guard cells. A T-DNA disruption mutation in AtP2C-HA confers ABA-hypersensitive regulation of stomatal closing and seed germination. The presented data provide a basis for cell type–specific genomic scale analyses of gene function. PMID:14973164

Effect of heat shock on ultrastructure and calcium distribution in Lavandula pinnata L. glandular trichomes.

Science.gov (United States)

Huang, S S; Kirchoff, B K; Liao, J P

2013-02-01

The effects of heat shock (HS) on the ultrastructure and calcium distribution of Lavandula pinnata secretory trichomes are examined using transmission electron microscopy and potassium antimonate precipitation. After 48-h HS at 40°C, plastids become distorted and lack stroma and osmiophilic deposits, the cristae of the mitochondria become indistinct, the endoplasmic reticulum acquires a chain-like appearance with ribosomes prominently attached to the lamellae, and the plasma and organelle membranes become distorted. Heat shock is associated with a decrease in calcium precipitates in the trichomes, while the number of precipitates increases in the mesophyll cells. Prolonged exposure to elevated calcium levels may be toxic to the mesophyll cells, while the lack of calcium in the glands cell may deprive them of the normal protective advantages of elevated calcium levels. The inequality in calcium distribution may result not only from uptake from the transpiration stream, but also from redistribution of calcium from the trichomes to the mesophyll cells.

Amorphous areas in the cytoplasm of Dendrobium tepal cells

Science.gov (United States)

van Doorn, Wouter G.; Kirasak, Kanjana; Ketsa, Saichol

2013-01-01

In Dendrobium flowers some tepal mesophyll cells showed cytoplasmic areas devoid of large organelles. Such amorphous areas comprised up to about 40% of the cross-section of a cell. The areas were not bound by a membrane. The origin of these areas is not known. We show data suggesting that they can be formed from vesicle-like organelles. The data imply that these organelles and other material become degraded inside the cytoplasm. This can be regarded as a form of autophagy. The amorphous areas became surrounded by small vacuoles, vesicles or double membranes. These seemed to merge and thereby sequester the areas. Degradation of the amorphous areas therefore seemed to involve macroautophagy. PMID:23823702

The ubiquitous presence of exopolygalacturonase in maize suggests a fundamental cellular function for this enzyme.

Science.gov (United States)

Dubald, M; Barakate, A; Mandaron, P; Mache, R

1993-11-01

Exopolygalacturonase (exoPG) is a pectin-degrading enzyme abundant in maize pollen. Using immunochemistry and in situ hybridization it is shown that in addition to its presence in pollen, exoPG is also present in sporophytic tissues, such as the tapetum and mesophyll cells. The enzyme is located in the cytoplasm of pollen and of some mesophyll cells. In other mesophyll cells, the tapetum and the pollen tube, exoPG is located in the cell wall. The measurement of enzyme activity shows that exoPG is ubiquitous in the vegetative organs. These results suggest a general function for exoPG in cell wall edification or degradation. ExoPG is encoded by a closely related multigene family. The regulation of the expression of one of the exoPG genes was analyzed in transgenic tobacco. Reporter GUS activity was detected in anthers, seeds and stems but not in leaves or roots of transgenic plants. This strongly suggests that the ubiquitous presence of exoPG in maize is the result of the expression of different exoPG genes.

Comparison of physiological and anatomical changes of C3 (Oryza sativa [L.]) and C4 (Echinochloa crusgalli [L.]) leaves in response to drought stress

Science.gov (United States)

Hamim, Hamim; Banon, Sri; Dorly, Dorly

2016-01-01

The experiment aimed to analyse the different response of C3 (Oryza sativa L.) and C4 (Echinochloa crusgalli L.) species to drought stress based on physiological and anatomical properties. Seeds of rice (Oryza sativa) and Echinochloa (Echinochloa crusgalli) were grown in 15 cm (D) pot for 6 weeks under well-watered conditions. After 6 weeks the plants were divided into two groups, (1) well-watered which were watered daily, and (2) drought stress which were withheld from watering for 6 days. After 6 days of drought, the plants were then re-watered to analyse plant recovery. During drought period, the plants were analysed for growth, leaf relative water content (RWC), photosynthesis, and leaf anatomy. Drought stress significantly reduced leaf RWC of both species, but the reduction was bigger in rice than in Echinochloa. The maximum efficiency of photosynthesis (Fv/Fm) was decrease significantly in response to drought stress by about 48.04% in rice, while it was only 34.40% in Echinochloa. Anatomical analysis showed drought treatment tended to reduce leaf thickness in the area of bulliform cell, major- as well as intervein and xylem diameter, more in Echinochloa than in rice, suggesting that the decrease of vein and xylem diameter is among the anatomical parameters that is important to overcome from drought stress in Echinochloa. The number of chloroplast in the mesophyll cell and bundle sheath cell (BSC) was different between these two species, where in Echinochloa chloroplast was found in both mesophyll as well as BSC, while in rice it was only found in mesophyll cell, confirmed that Echinochloa is a C4 and rice is a C3 species. Interestingly, in Echinochloa, the number of chloroplast was significantly increased due to drought stress in BSC, but not in mesophyll cell. The number of starch granules also dramatically increased in response to drought in the mesophyll cells of rice and Echinochloa, and in the bundle sheath cell of Echinochloa which indicate that C3

Differential activity of Plasma and Vacuolar Membrane Transporters contributes to Genotypic Differences in Salinity Tolerance in a Halophyte Species, Chenopodium quinoa

DEFF Research Database (Denmark)

Bonales-Alatorre, Edgar; Pottosin, Igor; Shabala, Lana

2013-01-01

quinoa genotypes contrasting in their salinity tolerance. The work is complemented by studies of the kinetics of net ion fluxes across the plasma membrane of quinoa leaf mesophyll tissue. Our results suggest that multiple mechanisms contribute towards genotypic differences in salinity tolerance in quinoa....... These include: (i) a higher rate of Na+ exclusion from leaf mesophyll; (ii) maintenance of low cytosolic Na+ levels; (iii) better K+ retention in the leaf mesophyll; (iv) a high rate of H+ pumping, which increases the ability of mesophyll cells to restore their membrane potential; and (v) the ability to reduce......Halophytes species can be used as a highly convenient model system to reveal key ionic and molecular mechanisms that confer salinity tolerance in plants. Earlier, we reported that quinoa (Chenopodium quinoa Willd.), a facultative C3 halophyte species, can efficiently control the activity of slow...

Incorporation of uridine-H3 into healthy and tobacco necrosis virus-infected mesophyll cells of Chenopodium amaranticolor

International Nuclear Information System (INIS)

Faccioli, G.; Rubies-Autonel, C.

1975-01-01

Tritiated uridine was selectively incorporated into the nucleus, nucleolus and cytoplasm of actinomycin D-treated Chenopodium amaranticolor cells locally infected with a strain of tobacco necrosis virus (TNV), 3 days after inoculation. Healthy cells did not show such an incorporation. Chloroplasts, in both types of cells, were free of label. Treatment with pancreatic ribonuclease removed the label completely in the majority of nuclei and nucleoli of infected cells. Since infectivity tests showed that AMD treatment increased virus multiplication by 10-12%, it is conceivable to think that the incorporation observed was due to virus synthesis. Preliminary infectivity experiments also showed that treatment of the cells with cycloheximide inhibited virus multiplication up to 80%, while chloramphenicol increased such multiplication. Our results lead to the conclusion that nucleus, nucleolus and cytoplasm but not chloroplasts are the sites involved in the synthesis of TNV. (orig.) [de

Vertical leaf mass per area gradient of mature sugar maple reflects both height-driven increases in vascular tissue and light-driven increases in palisade layer thickness.

Science.gov (United States)

Coble, Adam P; Cavaleri, Molly A

2017-10-01

A key trait used in canopy and ecosystem function modeling, leaf mass per area (LMA), is influenced by changes in both leaf thickness and leaf density (LMA = Thickness × Density). In tall trees, LMA is understood to increase with height through two primary mechanisms: (i) increasing palisade layer thickness (and thus leaf thickness) in response to light and/or (ii) reduced cell expansion and intercellular air space in response to hydrostatic constraints, leading to increased leaf density. Our objective was to investigate within-canopy gradients in leaf anatomical traits in order to understand environmental factors that influence leaf morphology in a sugar maple (Acer saccharum Marshall) forest canopy. We teased apart the effects of light and height on anatomical traits by sampling at exposed and closed canopies that had different light conditions at similar heights. As expected, palisade layer thickness responded strongly to cumulative light exposure. Mesophyll porosity, however, was weakly and negatively correlated with light and height (i.e., hydrostatic gradients). Reduced mesophyll porosity was not likely caused by limitations on cell expansion; in fact, epidermal cell width increased with height. Palisade layer thickness was better related to LMA, leaf density and leaf thickness than was mesophyll porosity. Vein diameter and fraction of vascular tissue also increased with height and LMA, density and thickness, revealing that greater investment in vascular and support tissue may be a third mechanism for increased LMA with height. Overall, decreasing mesophyll porosity with height was likely due to palisade cells expanding into the available air space and also greater investments in vascular and support tissue, rather than a reduction of cell expansion due to hydrostatic constraints. Our results provide evidence that light influences both palisade layer thickness and mesophyll porosity and indicate that hydrostatic gradients influence leaf vascular and support

Application of the comet assay in studies of programmed cell death (PCD in plants

Directory of Open Access Journals (Sweden)

Maria Charzyńska

2014-01-01

Full Text Available Programmed cell death (PCD in plants is an intensively investigated process. One of the main characteristics of PCD in both animal and plant organisms is the non-random, internucleosomal fragmentation of nuclear DNA, usually analysed using total DNA gel electrophoresis or TUNEL method. In this paper we present application of the "comet assay" (Single Cell Gel Electrophoresis for detection of nDNA degradation in studies of PCD during plant life cycle. We analyzed three types of tissue: anther tapetum, endosperm and mesophyll which were prepared in different ways to obtain a suspension of viable cells (without cell walls. The comet assay gives a possibility of examination of the nDNA degradation in individual cell. This method is significant for studies of the plant tissue differentiation and senescence especially in the cases when it is not possible to isolate large number of cells at the same developmental stage.

Elevated CO2 can modify the response to a water status gradient in a steppe grass: from cell organelles to photosynthetic capacity to plant growth.

Science.gov (United States)

Jiang, Yanling; Xu, Zhenzhu; Zhou, Guangsheng; Liu, Tao

2016-07-12

The atmospheric CO2 concentration is rising continuously, and abnormal precipitation may occur more frequently in the future. Although the effects of elevated CO2 and drought on plants have been well reported individually, little is known about their interaction, particularly over a water status gradient. Here, we aimed to characterize the effects of elevated CO2 and a water status gradient on the growth, photosynthetic capacity, and mesophyll cell ultrastructure of a dominant grass from a degraded grassland. Elevated CO2 stimulated plant biomass to a greater extent under moderate changes in water status than under either extreme drought or over-watering conditions. Photosynthetic capacity and stomatal conductance were also enhanced by elevated CO2 under moderate drought, but inhibited with over-watering. Severe drought distorted mesophyll cell organelles, but CO2 enrichment partly alleviated this effect. Intrinsic water use efficiency (WUEi) and total biomass water use efficiency (WUEt) were increased by elevated CO2, regardless of water status. Plant structural traits were also found to be tightly associated with photosynthetic potentials. The results indicated that CO2 enrichment alleviated severe and moderate drought stress, and highlighted that CO2 fertilization's dependency on water status should be considered when projecting key species' responses to climate change in dry ecosystems.

Anatomic and histochemical examinations for the clarification of the contribution of biotic agents to forest dieback

Energy Technology Data Exchange (ETDEWEB)

Fink, S; Horsch, F; Filby, G; Fund, N; Gross, S; Hanisch, B; Kilz, E; Seidel, A [comps.

1986-04-01

In yellowed needles of firs and spruces from forest decline areas in the Southern Black Forest frequently necrotic phloem could be found, while the mesophyll cells were still intact. This first led to the assumption of a possible participation of phloemspecific pathogens (viruses, MLO). Needles suffering from atmospheric pollutants in contrast showed necroses of mesophyll cells with largely intact phloem. Identical symptoms with collapsed phloem and intact mesophyll could be observed in spruces which showed typical apical yellowing of the needles after cultivation in magnesium-free hydroponic solution. The symptoms on the yellowed needles in the higher Black Forest can therefore conclusively be explained with the there observed magnesium-deficiency. Possible interrelationships between biotic pathogens and nutritional status of the trees are discussed. In a number of yellowed, but also some green needles, fungal hyphae could be observed in the microscopical sections, preferably in the intercellulars. The significance of these fungi will further be investigated.

The Evolutionary Basis of Naturally Diverse Rice Leaves Anatomy.

Directory of Open Access Journals (Sweden)

Jolly Chatterjee

Full Text Available Rice contains genetically and ecologically diverse wild and cultivated species that show a wide variation in plant and leaf architecture. A systematic characterization of leaf anatomy is essential in understanding the dynamics behind such diversity. Therefore, leaf anatomies of 24 Oryza species spanning 11 genetically diverse rice genomes were studied in both lateral and longitudinal directions and possible evolutionary trends were examined. A significant inter-species variation in mesophyll cells, bundle sheath cells, and vein structure was observed, suggesting precise genetic control over these major rice leaf anatomical traits. Cellular dimensions, measured along three growth axes, were further combined proportionately to construct three-dimensional (3D leaf anatomy models to compare the relative size and orientation of the major cell types present in a fully expanded leaf. A reconstruction of the ancestral leaf state revealed that the following are the major characteristics of recently evolved rice species: fewer veins, larger and laterally elongated mesophyll cells, with an increase in total mesophyll area and in bundle sheath cell number. A huge diversity in leaf anatomy within wild and domesticated rice species has been portrayed in this study, on an evolutionary context, predicting a two-pronged evolutionary pathway leading to the 'sativa leaf type' that we see today in domesticated species.

Compartmentation and equilibration of abscisic acid in isolated Xanthium cells

International Nuclear Information System (INIS)

Bray, E.A.; Zeevaart, J.A.D.

1986-01-01

The compartmentation of endogenous abscisic acid (ABA), applied (+/-)-[ 3 H]ABA, and (+/-)-trans-ABA was measured in isolated mesophyll cells of the Chicago strain of Xanthium strumarium L. The release of ABA to the medium in the presence or absence of DMSO was used to determine the equilibration of ABA in the cells. It was found that a greater percentage of the (+/-)-[ 3 H]ABA and the (+/-)-trans-ABA was released into the medium than of the endogenous ABA, indicating that applied ABA did not equilibrate with the endogenous material. Therefore, in further investigations only the compartmentation of endogenous ABA was studied. Endogenous ABA was released from Xanthium cells according to the pH gradients among the various cellular compartments. Thus, darkness, high external pH, KNO 2 , and drought-stress all increased the efflux of ABA from the cells. Efflux of ABA from the cells in the presence of 0.6 M mannitol occurred within 30 seconds, but only 8% of the endogenous material was released during the 20 minute treatment

Amino acid transport across the tonoplast of vacuoles isolated from barley mesophyll protoplasts: Uptake of alanine, leucine, and glutamine

International Nuclear Information System (INIS)

Dietz, K.J.; Jaeger, R.; Kaiser, G.; Martinoia, E.

1990-01-01

Mesophyll protoplasts from leaves of well-fertilized barley (Hordeum vulgare L.) plants contained amino acids at concentrations as high as 120 millimoles per liter. With the exception of glutamic acid, which is predominantly localized in the cytoplasm, a major part of all other amino acids was contained inside the large central vacuole. Alanine, leucine, and glutamine are the dominant vacuolar amino acids in barley. Their transport into isolated vacuoles was studied using 14 C-labeled amino acids. Uptake was slow in the absence of ATP. A three- to sixfold stimulation of uptake was observed after addition of ATP or adenylyl imidodiphosphate an ATP analogue not being hydrolyzed by ATPases. Other nucleotides were ineffective in increasing the rate of uptake. ATP-Stimulated amino acid transport was not dependent on the transtonoplast pH or membrane potential. p-Chloromercuriphenylsulfonic acid and n-ethyl maleimide increased transport independently of ATP. Neutral amino acids such as valine or leucine effectively decreased the rate of alanine transport. Glutamine and glycine were less effective or not effective as competitive inhibitors of alanine transport. The results indicate the existence of a uniport translocator specific for neutral or basic amino acids that is under control of metabolic effectors

Asymmetrical effects of mesophyll conductance on fundamental photosynthetic parameters and their relationships estimated from leaf gas exchange measurements.

Science.gov (United States)

Sun, Ying; Gu, Lianhong; Dickinson, Robert E; Pallardy, Stephen G; Baker, John; Cao, Yonghui; DaMatta, Fábio Murilo; Dong, Xuejun; Ellsworth, David; Van Goethem, Davina; Jensen, Anna M; Law, Beverly E; Loos, Rodolfo; Martins, Samuel C Vitor; Norby, Richard J; Warren, Jeffrey; Weston, David; Winter, Klaus

2014-04-01

Worldwide measurements of nearly 130 C3 species covering all major plant functional types are analysed in conjunction with model simulations to determine the effects of mesophyll conductance (g(m)) on photosynthetic parameters and their relationships estimated from A/Ci curves. We find that an assumption of infinite g(m) results in up to 75% underestimation for maximum carboxylation rate V(cmax), 60% for maximum electron transport rate J(max), and 40% for triose phosphate utilization rate T(u) . V(cmax) is most sensitive, J(max) is less sensitive, and T(u) has the least sensitivity to the variation of g(m). Because of this asymmetrical effect of g(m), the ratios of J(max) to V(cmax), T(u) to V(cmax) and T(u) to J(max) are all overestimated. An infinite g(m) assumption also limits the freedom of variation of estimated parameters and artificially constrains parameter relationships to stronger shapes. These findings suggest the importance of quantifying g(m) for understanding in situ photosynthetic machinery functioning. We show that a nonzero resistance to CO2 movement in chloroplasts has small effects on estimated parameters. A non-linear function with gm as input is developed to convert the parameters estimated under an assumption of infinite gm to proper values. This function will facilitate gm representation in global carbon cycle models. © 2013 John Wiley & Sons Ltd.

Macroinvertebrates associated with two submerged macrophytes ...

African Journals Online (AJOL)

Macroinvertebrates associated with two submerged macrophytes, Lagarosiphon ilicifolius and Vallisneria aethiopica , in the Sanyati Basin, Lake Kariba, Zimbabwe: effect of plant morphological complexity.

Comparison of foliar anatomy of ten bread wheat (triticum, poaceae) and ten barley (hordeum, poaceae) cultivars

International Nuclear Information System (INIS)

Ardic, M.; Sezer, O.; Ozgdsd, K.; Yaylaci, O. K.; Koyuncu, O.; Olgun, M.; Bascdftcd, Z. B.; Ayter, N. G.

2015-01-01

The aim of this study is to determine anatomical differences and classification of leaf and leaf cell characteristics (cuticle thickness, upper epidermis thickness, lower epidermis thickness, mesophyll thickness, parenchyma thickness and leaf thickness) between 10 bread wheat cultivars (Triticum aestivum L.) and 10 barley cultivars (Hordeum vulgare L.). Classification of leaf characteristics in bread wheat and barley cultivars and relationship between leaf characteristics are made by principal component and correlation analyses. Highest thickness belongs to W8 Mufitbey cultivar in mesophyll and lower epidermis and W1 Sonmez 01 cultivar have the lowest thickness of upper epidermis in bread wheat. In Barley, B1 Ince cultivar has highest leaf thickness mesophyll and parenchyma; lowest thickness of cuticle is included B7 Cumhuriyet 50 cultivar. All other cultivars have homogenous contents of leaf characteristics. (author)

Measurement of gross photosynthesis, respiration in the light, and mesophyll conductance using H218O labeling.

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Gauthier, Paul Pg; Battle, Mark O; Griffin, Kevin L; Bender, Michael L

2018-03-27

A fundamental challenge in plant physiology is independently determining the rates of gross O2 production by photosynthesis and O2 consumption by respiration, photorespiration, and other processes. Previous studies on isolated chloroplasts or leaves have separately constrained net and gross O2 production (NOP and GOP, respectively) by labeling ambient O2 with 18O while leaf water was unlabeled. Here, we describe a method to accurately measure GOP and NOP of whole detached leaves in a cuvette as a routine gas exchange measurement. The petiole is immersed in water enriched to a δ18O of ~9,000‰, and leaf water is labeled through the transpiration stream. Photosynthesis transfers 18O from H2O to O2. GOP is calculated from the increase in δ18O of O2 as air passes through the cuvette. NOP is determined from the increase in O2/N2. Both terms are measured by isotope ratio mass spectrometry. CO2 assimilation and other standard gas exchange parameters are also measured. Reproducible measurements are made on a single leaf for more than 15 hours. We used this method to measure the light response curve of NOP and GOP in Phaseolus vulgaris at 21% and 2% O2. We then used these data to examine the O2/CO2 ratio of net photosynthesis, the light response curve of mesophyll conductance, and the apparent inhibition of respiration in the light (Kok effect) at both oxygen levels. The results are discussed in the context of evaluating the technique as a tool to study and understand leaf physiological traits. {copyright, serif} 2018 American Society of Plant Biologists. All rights reserved.

Influence of Drought on Mesophyll Resistance to CO2 Diffusion and its Impact on Water-Use Efficiency in Trees

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Guo, J.; Beverly, D.; Cook, C.; Ewers, B. E.; Williams, D. G.

2015-12-01

The resistance to CO2 diffusion inside leaves (mesophyll resistance; rm) during photosynthesis is often comparable in magnitude to stomatal diffusion resistance, and varies among species and across environmental conditions. Consequently, photosynthesis is strongly limited by rm at low internal CO2 partial pressures, such that its variation may determine patterns of leaf water-use efficiency (WUE). Reduction in stomatal conductance with drought typically increases WUE, but also decreases photosynthesis. In theory, the decrease in photosynthesis could be countered by reduction in rm while maintaining high WUE. It is still uncertain how drought-related changes in rm affect short- and long-term WUE strategies of different tree species. We conducted field observations of instantaneous WUE and 13C discrimination in two dominant conifer species (Pinus contorta and Picea engelmannii) in SE Wyoming over the seasonal dry-down period in the summer of 2015. rm was examined by on-line 13C discrimination using isotope laser spectroscopy. Controlled environment studies on three conifer species (P. contorta, P. engelmannii, and Abies lasiocarpa) and one angiosperm (Populus tremuloides) are in progress. We hypothesize that the plasticity of rm in response to drought accounts for significant adjustments in photosynthetic capacity and WUE. Needle leaf conifers are known to have relatively high rm, and we expect them to show greater improvements in photosynthesis and WUE when rm is decreased compared to angiosperm tree species.

Comparative proteomics of chloroplasts envelopes from bundle sheath and mesophyll chloroplasts reveals novel membrane proteins with a possible role in C4-related metabolite fluxes and development.

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Kalpana eManandhar-Shrestha

2013-03-01

Full Text Available As the world population grows, our need for food increases drastically. Limited amounts of arable land lead to a competition between food and fuel crops, while changes in the global climate may impact future crop yields. Thus, a second green revolution will need a better understanding of the processes essential for plant growth and development. One approach toward the solution of this problem is to better understand regulatory and transport processes in C4 plants. C4 plants display an up to 10-fold higher apparent CO2 assimilation and higher yields while maintaining high water use efficiency. This requires differential regulation of mesophyll (M and bundle sheath (BS chloroplast development as well as higher metabolic fluxes of photosynthetic intermediates between cells and across chloroplast envelopes. While previous analyses of overall chloroplast membranes have yielded significant insight, our comparative proteomics approach using enriched BS and M chloroplast envelopes of Zea mays allowed us to identify 37 proteins of unknown function that have not been seen in these earlier studies. We identified 280 proteins, 84% of which are known/predicted to be present in chloroplasts (cp. 74% have a known or predicted membrane association. 21 membrane proteins were 2-15 times more abundant in BS cells, while 36 proteins were more abundant in M cp envelopes. These proteins could represent additional candidates of proteins essential for development or metabolite transport processes in C4 plants. RT-PCR confirmed differential expression of thirteen candidate genes. Cp association was confirmed using GFP labeling. Genes for a PIC-like protein and an ER-AP-like protein show an early transient increase in gene expression during the transition to light. In addition, PIC gene expression is increased in the immature part of the leaf and was lower in the fully developed parts of the leaf, suggesting a need for/incorporation of the protein during chloroplast

A two-dimensional microscale model of gas exchange during photosynthesis in maize (Zea mays L.) leaves.

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Retta, Moges; Ho, Quang Tri; Yin, Xinyou; Verboven, Pieter; Berghuijs, Herman N C; Struik, Paul C; Nicolaï, Bart M

2016-05-01

CO2 exchange in leaves of maize (Zea mays L.) was examined using a microscale model of combined gas diffusion and C4 photosynthesis kinetics at the leaf tissue level. Based on a generalized scheme of photosynthesis in NADP-malic enzyme type C4 plants, the model accounted for CO2 diffusion in a leaf tissue, CO2 hydration and assimilation in mesophyll cells, CO2 release from decarboxylation of C4 acids, CO2 fixation in bundle sheath cells and CO2 retro-diffusion from bundle sheath cells. The transport equations were solved over a realistic 2-D geometry of the Kranz anatomy obtained from light microscopy images. The predicted responses of photosynthesis rate to changes in ambient CO2 and irradiance compared well with those obtained from gas exchange measurements. A sensitivity analysis showed that the CO2 permeability of the mesophyll-bundle sheath and airspace-mesophyll interfaces strongly affected the rate of photosynthesis and bundle sheath conductance. Carbonic anhydrase influenced the rate of photosynthesis, especially at low intercellular CO2 levels. In addition, the suberin layer at the exposed surface of the bundle sheath cells was found beneficial in reducing the retro-diffusion. The model may serve as a tool to investigate CO2 diffusion further in relation to the Kranz anatomy in C4 plants. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Electron-cytochemical study of Ca2+ in cotyledon cells of soybean seedlings grown in microgravity

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Nedukha, O.; Brown, C. S.; Kordyum, E.; Piastuch, W. C.; Guikema, J. A. (Principal Investigator)

1999-01-01

Microgravity and horizontal clinorotation are known to cause the rearrangement of the structural-functional organization of plant cells, leading to accelerated aging. Altered gravity conditions resulted in an increase in the droplets volume in cells and the destruction of chloroplast structure in Arabidopsis thaliana plants, an enhancement of cytosolic autophagaous processes, an increase in the respiration rate and a greater number of multimolecular forms of succinate- and malate dehydrogenases in cells of the Funaria hygrometrica protonema and Chlorella vulgaris, and changes in calcium balance of cells. Because ethylene is known to be involved in cell aging and microgravity appears to speed the process, and because soybean seedlings grown in space produce higher ethylene levels we asked: 1) does an acceleration of soybean cotyledon cell development and aging occur in microgravity? 2) what roles do Ca2+ ions and the enhanced ethylene level play in these events? Therefore, the goal of our investigation was to examine of the interaction of microgravity and ethylene on the localization of Ca2+ in cotyledon mesophyll of soybean seedlings.

Analyses of Aloe polysaccharides using carbohydrate microarray profiling

DEFF Research Database (Denmark)

Isager Ahl, Louise; Grace, Olwen M; Pedersen, Henriette Lodberg

2018-01-01

As the popularity of Aloe vera extracts continues to rise, a desire to fully understand the individual polymer components of the leaf mesophyll, their relation to one another and the effects they have on the human body are increasing. Polysaccharides present in the leaf mesophyll have been...... identified as the components responsible for the biological activities of Aloe vera, and they have been widely studied in the past decades. However, the commonly used methods do not provide the desired platform to conduct large comparative studies of polysaccharide compositions as most of them require...... a complete or near-complete fractionation of the polymers. The objective for this study was to assess whether carbohydrate microarrays could be used for the high-throughput analysis of cell wall polysaccharides in Aloe leaf mesophyll. The method we chose is known as Comprehensive Microarray Polymer Profiling...

Physical Changes in Satsuma Mandarin Leaf after Infection of Elsinoë fawcettii Causing Citrus Scab Disease

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Dilli Prasad Paudyal

2015-12-01

Full Text Available Citrus scab disease is one of the destructive diseases that reduce the value of fruit for the fresh market. We analyzed the process of symptom development after infection with scab pathogen Elsinoë fawcettii in the susceptible satsuma mandarin leaves to observe the structural modification against pathogen. The cuticle and epidermal cells along with 3–5 layers of mesophyll tissue were degraded 1–2 days post inoculation. Surrounding peripheral cells of degraded tissues grew rapidly and then enveloped the necrotic area along with the growing conidia. Cross sections through the lesion revealed hyphal colonization in epidermis and mesophyll tissues. In response to the pathogen colonization, host cell walls were lignified, inner cells were rapidly compartmentalized and a semi-circular boundary was formed that separated the infected region from the non-infected region, and finally prevented the intercellular pathogen spread.

Effects of the harmful gases SO/sub 2/ and HF on plant leaf structure

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Qin, H; Wu, Z; Wang, J; Qian, D; Li, Z

1980-09-01

The injury induced by SO/sub 2/ appeared progressively; cells contracted and became deformed, the protoplasm and the chloroplasts turned yellow-brown or collapsed while no effects were seen in the vascular bundles. However, the injury induced by HF were different; the cells were not deformed immediately, the protoplasm became red-brown, the mesophyll cells adjacent to stomata or vascular bundles became red-brown too, and there were no effects on chloroplasts, which did not collapse until the tissue necrosis appeared. The cells of xylem and phloem turned red-brown. The process of injury to leaf structure induced by SO/sub 2/ is discussed. It is observed that destruction of chlorophyll and the interruption of photosynthesis by SO/sub 2/ took place first in the palisade tissue, whereas the contraction and disintegration of the cells happened first in the spongy tissue. The effect of HF (the contractive collapse of chlorophyll and mesophyll) occurred after the influence on protoplasm appeared.

Greater efficiency of water use in poplar clones having a delayed response of mesophyll conductance to drought.

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Théroux Rancourt, Guillaume; Éthier, Gilbert; Pepin, Steeve

2015-02-01

Improvement of water use efficiency is a key objective to improve the sustainability of cultivated plants, especially fast growing species with high water consumption like poplar. It is well known that water use efficiency (WUE) varies considerably among poplar genotypes, and it was recently suggested that the use of the mesophyll-to-stomatal conductance ratio (gm/gs) would be an appropriate trait to improve WUE. The responses of 7-week-old cuttings of four hybrid poplar clones and one native Balsam poplar (Populus balsamifera L.) to a water stress-recovery cycle were examined to evaluate the relation between the gm/gs ratio and transpiration efficiency (TE), a leaf-level component of WUE. A contrasting gs response to water stress was observed among the five clones, from stomatal closure early on during soil drying up to limited closure in Balsam poplar. However in the hybrids, the decline in gm was consistently delayed by a few days compared with gs. Moreover, in the most water use-efficient hybrids, the recovery following rehydration occurred faster for gm than for gs. Thus, the delay in the response of gm to drought and its faster recovery upon rewatering increased the gm/gs of the hybrids and this ratio scaled positively with TE. Our results support the use of the gm/gs ratio to select genotypes with improved WUE, and the notion that breeding strategies focusing mainly on stomatal responses to soil drying should also look for a strong curvilinearity between net carbon assimilation rate and gs, the indication of a significant increase in gm/gs in the earlier stages of stomatal closure. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Enhanced phosphorus reduction in simulated eutrophic water: a comparative study of submerged macrophytes, sediment microbial fuel cells, and their combination.

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Xu, Peng; Xiao, Enrong; Xu, Dan; Li, Juan; Zhang, Yi; Dai, Zhigang; Zhou, Qiaohong; Wu, Zhenbin

2018-05-01

The phosphorus reduction in water column was attempted by integrating sediment microbial fuel cells (SMFCs) with the submerged macrophyte Vallisneria spiralis. A comparative study was conducted to treat simulated water rich in phosphate with a control and three treatments: SMFC alone (SMFC), submerged macrophytes alone (macophyte), and combined macrophytes and fuel cells (M-SMFC). All treatments promoted phosphorus flux from the water column to sediments. Maximum phosphorus reduction was obtained in proportion to the highest stable phosphorus level in sediments in M-SMFC. For the initial phosphate concentrations of 0.2, 1, 2, and 4 mg/L, average phosphate values in the overlying water during four phases decreased by 33.3% (25.0%, 8.3%), 30.8% (5.1%, 17.9%), 36.5% (27.8%, 15.7%), and 36.2% (0.7%, 22.1%) for M-SMFC (macrophyte, SMFC), compared with the control. With macrophyte treatment, the obvious phosphorus release from sediments was observed during the declining period. However, such phenomenon was significantly inhibited with M-SMFC. The electrogenesis bacteria achieved stronger phosphorus adsorption and assimilation was significantly enriched on the closed-circuit anodes. The higher abundance of Geobacter and Pseudomonas in M-SMFC might in part explain the highest phosphorus reduction in the water column. M-SMFC treatment could be promising to control the phosphorus in eutrophic water bodies.

Light-regulated leaf expansion in two Populus species: dependence on developmentally controlled ion transport.

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Stiles, Kari A; Van Volkenburgh, Elizabeth

2002-07-01

Leaf growth responses to light have been compared in two species of Populus, P. deltoides and P. trichocarpa. These species differ markedly in morphology, anatomy, and dependence on light during leaf expansion. Light stimulates the growth rate and acidification of cell walls in P. trichocarpa but not in P. deltoides, whereas leaves of P. deltoides maintain growth in the dark. Light-induced growth is promoted in P. deltoides when cells are provided 50-100 mM KCl. In both species, light initially depolarizes, then hyperpolarizes mesophyll plasma membranes. However, in the dark, the resting E(m) of mesophyll cells in P. deltoides, but not in P. trichocarpa, is relatively insensitive to decade changes in external [K+]. Results suggest that light-stimulated leaf growth depends on developmentally regulated cellular mechanisms controlling ion fluxes across the plasma membrane. These developmental differences underlie species-level differences in growth and physiological responses to the photoenvironment.

Vesicles between plasma membrane and cell wall prior to visible senescence of Iris and Dendrobium flowers.

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Kamdee, Channatika; Kirasak, Kanjana; Ketsa, Saichol; van Doorn, Wouter G

2015-09-01

Cut Iris flowers (Iris x hollandica, cv. Blue Magic) show visible senescence about two days after full opening. Epidermal cells of the outer tepals collapse due to programmed cell death (PCD). Transmission electron microscopy (TEM) showed irregular swelling of the cell walls, starting prior to cell collapse. Compared to cells in flowers that had just opened, wall thickness increased up to tenfold prior to cell death. Fibrils were visible in the swollen walls. After cell death very little of the cell wall remained. Prior to and during visible wall swelling, vesicles (paramural bodies) were observed between the plasma membrane and the cell walls. The vesicles were also found in groups and were accompanied by amorphous substance. They usually showed a single membrane, and had a variety of diameters and electron densities. Cut Dendrobium hybrid cv. Lucky Duan flowers exhibited visible senescence about 14 days after full flower opening. Paramural bodies were also found in Dendrobium tepal epidermis and mesophyll cells, related to wall swelling and degradation. Although alternative explanations are well possible, it is hypothesized that paramural bodies carry enzymes involved in cell wall breakdown. The literature has not yet reported such bodies in association with senescence/PCD. Copyright © 2015 Elsevier GmbH. All rights reserved.

Sinopsis sobre las Hydrocharitaceae de Colombia

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Schmidt Mumm Udo

1996-06-01

Full Text Available The aquatic family Hydrocharitaceae is actually represented in Colombia by seven genera and seven species (Egeria densa, Apalanthe granatensis, Halophila decipiens, Hydrilla verticillata, Limnobium laevigatum, Thalassia testudinum, Vallisneria americana. A first record of Hydrilla verticillata growing spontaneously in outside conditions is made for this geographical area. A key to genera is presented, with each species briefly commented and illustrated.En Colombia la familia de plantas acuáticas Hydrocharitaceae se encuentra representada por siete géneros y siete especies (Egeria densa, Apalanthe granatensis, Halophila decipiens, Hydrilla verticillata, Limnabium laevigatum, Thalassia testudinum, Vallisneria americana. Para el país se registra por primera vez a Hydrilla verticillata. Adicionalmente se presenta una clave para los géneros, con las especies brevemente comentadas e ilustradas.

Effect of Hf on the fine structure of mesophyll cells from Glycine max, Merr

Energy Technology Data Exchange (ETDEWEB)

Wei, L.; Miller, G.W.

1972-04-01

A series of ultrastructural changes were observed in soybean leaves fumigated with 40 to 50 ppb of hydrogen fluoride. In the cytoplasm the presence of small vacuoles was the first noticeable initial change. The fragmentation of the vacuolar membrane occurred either simultaneously or followed immediately. Lipid-droplet-like globules and numerous vesicles occurred subsequently in the cytoplasm and increased as the injury became more severe. There was a decrease in polysomes and a detachment of ribosome from the rough endoplasmic reticulum. Free ribosome concentration also decreased as the injury became severe. Mitochondrial modifications involving dilation of outer and cristae membranes followed by reduction of both cristae number and matrix electron density and the disappearance of mitochondrial granules were observed in the chlorotic leaves. Electron dense inclusions accumulated in some mitochondria as well. The first noticeable change observed in the chloroplast was the presence of clusters of phytoferritin granules within the stoma after only 2 days of fumigation. Alterations in nuclear structures were observed in later stages of injury. Numerous small electron dense particles were found on various types of membranes in cells of severely chlorotic leaves. They were distributed on outer mitochondrial membranes, endoplasmic reticula, dictyosomes, tonoplasts, plasmalemma, nuclear envelopes, and disintegrating organelles and vesicles, but were never observed on membranes of chloroplasts and microbodies. The presence of fluoride has attracted the attention of many workers primarily in certain industrial areas where the emitted atmospheric fluoride concentrates and is accumulated by plants initiating injury. 6 references.

Mesophyll conductance decreases in the wild type but not in an ABA-deficient mutant (aba1) of Nicotiana plumbaginifolia under drought conditions.

Science.gov (United States)

Mizokami, Yusuke; Noguchi, Ko; Kojima, Mikiko; Sakakibara, Hitoshi; Terashima, Ichiro

2015-03-01

Under drought conditions, leaf photosynthesis is limited by the supply of CO2 . Drought induces production of abscisic acid (ABA), and ABA decreases stomatal conductance (gs ). Previous papers reported that the drought stress also causes the decrease in mesophyll conductance (gm ). However, the relationships between ABA content and gm are unclear. We investigated the responses of gm to the leaf ABA content [(ABA)L ] using an ABA-deficient mutant, aba1, and the wild type (WT) of Nicotiana plumbaginifolia. We also measured leaf water potential (ΨL ) because leaf hydraulics may be related to gm . Under drought conditions, gm decreased with the increase in (ABA)L in WT, whereas both (ABA)L and gm were unchanged by the drought treatment in aba1. Exogenously applied ABA decreased gm in both WT and aba1 in a dose-dependent manner. ΨL in WT was decreased by the drought treatment to -0.7 MPa, whereas ΨL in aba1 was around -0.8 MPa even under the well-watered conditions and unchanged by the drought treatment. From these results, we conclude that the increase in (ABA)L is crucial for the decrease in gm under drought conditions. We discuss possible relationships between the decrease in gm and changes in the leaf hydraulics. © 2014 John Wiley & Sons Ltd.

Leaf structural adaptations of two Limonium miller (Plumbaginales, Plumbaginaceae taxa

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Zorić Lana N.

2013-01-01

Full Text Available Limonium gmelinii (Willd. O. Kuntze 1891 subsp. hungaricum (Klokov Soó is Pannonian endemic subspecies that inhabits continental halobiomes, while Limonium anfractum (Salmon Salmon 1924 is one of the indicators of halophyte vegetation of marine rocks and its distribution is restricted to the southern parts of Mediterranean Sea coast. In this work, micromorphological and anatomical characters of leaves of these two Limonium taxa were analyzed, in order to examine their adaptations to specific environmental conditions on saline habitats. The results showed that both taxa exhibited strong xeromorphic adaptations that reflected in flat cell walls of epidermal cells, thick cuticle, high palisade/spongy tissue ratio, high index of palisade cells, the presence of sclereid idioblasts in leaf mesophyll and mechanical tissue by phloem and xylem. Both taxa are crynohalophytes and have salt glands on adaxial and abaxial epidermis for excretion of surplus salt. Relatively high dimensions of mesophyll cells, absence of non-glandular hairs and unprotected stomata slightly increased above the level of epidermal cells, are also adaptations to increased salinity. [Projekat Ministarstva nauke Republike Srbije, br. 173002

An update: improvements in imaging perfluorocarbon-mounted plant leaves with implications for studies of plant pathology, physiology, development and cell biology.

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Littlejohn, George R; Mansfield, Jessica C; Christmas, Jacqueline T; Witterick, Eleanor; Fricker, Mark D; Grant, Murray R; Smirnoff, Nicholas; Everson, Richard M; Moger, Julian; Love, John

2014-01-01

Plant leaves are optically complex, which makes them difficult to image by light microscopy. Careful sample preparation is therefore required to enable researchers to maximize the information gained from advances in fluorescent protein labeling, cell dyes and innovations in microscope technologies and techniques. We have previously shown that mounting leaves in the non-toxic, non-fluorescent perfluorocarbon (PFC), perfluorodecalin (PFD) enhances the optical properties of the leaf with minimal impact on physiology. Here, we assess the use of the PFCs, PFD, and perfluoroperhydrophenanthrene (PP11) for in vivo plant leaf imaging using four advanced modes of microscopy: laser scanning confocal microscopy (LSCM), two-photon fluorescence microscopy, second harmonic generation microscopy, and stimulated Raman scattering (SRS) microscopy. For every mode of imaging tested, we observed an improved signal when leaves were mounted in PFD or in PP11, compared to mounting the samples in water. Using an image analysis technique based on autocorrelation to quantitatively assess LSCM image deterioration with depth, we show that PP11 outperformed PFD as a mounting medium by enabling the acquisition of clearer images deeper into the tissue. In addition, we show that SRS microscopy can be used to image PFCs directly in the mesophyll and thereby easily delimit the "negative space" within a leaf, which may have important implications for studies of leaf development. Direct comparison of on and off resonance SRS micrographs show that PFCs do not to form intracellular aggregates in live plants. We conclude that the application of PFCs as mounting media substantially increases advanced microscopy image quality of living mesophyll and leaf vascular bundle cells.

An update: improvements in imaging perfluorocarbon-mounted plant leaves with implications for studies of plant pathology, physiology, development and cell biology.

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George R Littlejohn

2014-04-01

Full Text Available Plant leaves are optically complex, which makes them difficult to image by light microscopy. Careful sample preparation is therefore required to enable researchers to maximise the information gained from advances in fluorescent protein labelling, cell dyes and innovations in microscope technologies and techniques. We have previously shown that mounting leaves in the non-toxic, non-fluorescent perfluorocarbon (PFC, perfluorodecalin (PFD enhances the optical properties of the leaf with minimal impact on physiology. Here, we assess the use of the perfluorocarbons PFD, and perfluoroperhydrophenanthrene (PP11 for in vivo plant leaf imaging using 4 advanced modes of microscopy: laser scanning confocal microscopy (LSCM, Two-photon fluorescence (TPF microscopy, second harmonic generation (SHG microscopy and stimulated Raman scattering (SRS microscopy. For every mode of imaging tested, we observed an improved signal when leaves were mounted in PFD or in PP11, compared to mounting the samples in water. Using an image analysis technique based on autocorrelation to quantitatively assess LSCM image deterioration with depth, we show that PP11 outperformed PFD as a mounting medium by enabling the acquisition of clearer images deeper into the tissue. In addition, we show that SRS microscopy can be used to image perfluorocarbons directly in the mesophyll and thereby easily delimit the negative space within a leaf, which may have important implications for studies of leaf development. Direct comparison of on and off resonance SRS micrographs show that PFCs do not to form intracellular aggregates in live plants. We conclude that the application of PFCs as mounting media substantially increases advanced microscopy image quality of living mesophyll and leaf vascular bundle cells.

Ultrastructure of the Rust Fungus Puccinia miscanthi in the Teliospore Stage Interacting with the Biofuel Plant Miscanthus sinensis

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Ki Woo Kim

2015-09-01

Full Text Available Interaction of the the rust fungus Puccinia miscanthi with the biofuel plant Miscanthus sinensis during the teliospore phase was investigated by light and electron microscopy. P. miscanthi telia were oval-shaped and present on both the adaxial and abaxial leaf surfaces. Teliospores were brown, one-septate (two-celled, and had pedicels attached to one end. Transmission electron microscopy revealed numerous electron-translucent lipid globules in the cytoplasm of teliospores. Extensive cell wall dissolution around hyphae was not observed in the host tissues beneath the telia. Hyphae were found between mesophyll cells in the leaf tissues as well as in host cells. Intracellular hyphae, possibly haustoria, possessed electron-dense fungal cell walls encased by an electron-transparent fibrillar extrahaustorial sheath that had an electron-dense extrahaustorial membrane. The infected host cells appeared to maintain their membrane-bound structures such as nuclei and chloroplasts. These results suggest that the rust fungus maintains its biotrophic phase with most mesophyll cells of M. sinensis. Such a nutritional mode would permit the rust fungus to obtain food reserves for transient growth in the course of host alteration.

Effects of elevated ozone on CO2 uptake and leaf structure in sugar maple under two light environments

International Nuclear Information System (INIS)

Bäck, J.; Vanderklein, D.W.; Topa, M.A.

1999-01-01

The interactive effects of ozone and light on leaf structure, carbon dioxide uptake and short-term carbon allocation of sugar maple (Acer saccharum Marsh.) seedlings were examined using gas exchange measurements and 14 C-macroautoradiographic techniques. Two-year-old sugar maple seedlings were fumigated from budbreak for 5 months with ambient or 3 × ambient ozone in open-top chambers, receiving either 35% (high light) or 15% (low light) of full sunlight. Ozone accelerated leaf senescence, and reduced net photosynthesis, 14 CO 2 uptake and stomatal conductance, with the effects being most pronounced under low light. The proportion of intercellular space increased in leaves of seedlings grown under elevated ozone and low light, possibly enhancing the susceptibility of mesophyll cells to ozone by increasing the cumulative dose per mesophyll cell. Indeed, damage to spongy mesophyll cells in the elevated ozone × low light treatment was especially frequent. 14 C macroautoradioraphy revealed heterogeneous uptake of 14 CO 2 in well defined areole regions, suggesting patchy stomatal behaviour in all treatments. However, in seedlings grown under elevated ozone and low light, the highest 14 CO 2 uptake occurred along larger veins, while interveinal regions exhibited little or no uptake. Although visible symptoms of ozone injury were not apparent in these seedlings, the cellular damage, reduced photosynthetic rates and reduced whole-leaf chlorophyll levels corroborate the visual scaling of whole-plant senescence, suggesting that the ozone × low light treatment accelerated senescence or senescence-like injury in sugar maple. (author)

David et al (12)

African Journals Online (AJOL)

DELL

larger mesophyll cells, xylem tissues, vascular bundles and bundle sheaths than other cultivars. Thus, larger ... mechanism to cope with drought stress. There was an ... where solar radiation is intense (Løe et al., 2007). Most ecological studies ...

Differential Activity of Plasma and Vacuolar Membrane Transporters Contributes to Genotypic Differences in Salinity Tolerance in a Halophyte Species, Chenopodium quinoa

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Edgar Bonales-Alatorre

2013-04-01

Full Text Available Halophytes species can be used as a highly convenient model system to reveal key ionic and molecular mechanisms that confer salinity tolerance in plants. Earlier, we reported that quinoa (Chenopodium quinoa Willd., a facultative C3 halophyte species, can efficiently control the activity of slow (SV and fast (FV tonoplast channels to match specific growth conditions by ensuring that most of accumulated Na+ is safely locked in the vacuole (Bonales-Alatorre et al. (2013 Plant Physiology. This work extends these finding by comparing the properties of tonoplast FV and SV channels in two quinoa genotypes contrasting in their salinity tolerance. The work is complemented by studies of the kinetics of net ion fluxes across the plasma membrane of quinoa leaf mesophyll tissue. Our results suggest that multiple mechanisms contribute towards genotypic differences in salinity tolerance in quinoa. These include: (i a higher rate of Na+ exclusion from leaf mesophyll; (ii maintenance of low cytosolic Na+ levels; (iii better K+ retention in the leaf mesophyll; (iv a high rate of H+ pumping, which increases the ability of mesophyll cells to restore their membrane potential; and (v the ability to reduce the activity of SV and FV channels under saline conditions. These mechanisms appear to be highly orchestrated, thus enabling the remarkable overall salinity tolerance of quinoa species.

Differential activity of plasma and vacuolar membrane transporters contributes to genotypic differences in salinity tolerance in a Halophyte Species, Chenopodium quinoa.

Science.gov (United States)

Bonales-Alatorre, Edgar; Pottosin, Igor; Shabala, Lana; Chen, Zhong-Hua; Zeng, Fanrong; Jacobsen, Sven-Erik; Shabala, Sergey

2013-04-29

Halophytes species can be used as a highly convenient model system to reveal key ionic and molecular mechanisms that confer salinity tolerance in plants. Earlier, we reported that quinoa (Chenopodium quinoa Willd.), a facultative C3 halophyte species, can efficiently control the activity of slow (SV) and fast (FV) tonoplast channels to match specific growth conditions by ensuring that most of accumulated Na+ is safely locked in the vacuole (Bonales-Alatorre et al. (2013) Plant Physiology). This work extends these finding by comparing the properties of tonoplast FV and SV channels in two quinoa genotypes contrasting in their salinity tolerance. The work is complemented by studies of the kinetics of net ion fluxes across the plasma membrane of quinoa leaf mesophyll tissue. Our results suggest that multiple mechanisms contribute towards genotypic differences in salinity tolerance in quinoa. These include: (i) a higher rate of Na+ exclusion from leaf mesophyll; (ii) maintenance of low cytosolic Na+ levels; (iii) better K+ retention in the leaf mesophyll; (iv) a high rate of H+ pumping, which increases the ability of mesophyll cells to restore their membrane potential; and (v) the ability to reduce the activity of SV and FV channels under saline conditions. These mechanisms appear to be highly orchestrated, thus enabling the remarkable overall salinity tolerance of quinoa species.

The influences of Hygromycin B on growth of Arabidopsis thaliana ...

African Journals Online (AJOL)

use

2011-12-05

Dec 5, 2011 ... culture time, cells in the mesophyll tissue of cotyledon were loose arranged and the intercellular space was large. ... surface to avoid strong solar burn (Liu and Teskey, 1995; .... In order to further study functional mechanism of.

The role of mesophyll conductance during water stress and recovery in tobacco (Nicotiana sylvestris): acclimation or limitation?

Science.gov (United States)

Galle, Alexander; Florez-Sarasa, Igor; Tomas, Magdalena; Pou, Alicia; Medrano, Hipolito; Ribas-Carbo, Miquel; Flexas, Jaume

2009-01-01

While the responses of photosynthesis to water stress have been widely studied, acclimation to sustained water stress and recovery after re-watering is poorly understood. In particular, the factors limiting photosynthesis under these conditions, and their possible interactions with other environmental conditions, are unknown. To assess these issues, changes of photosynthetic CO(2) assimilation (A(N)) and its underlying limitations were followed during prolonged water stress and subsequent re-watering in tobacco (Nicotiana sylvestris) plants growing under three different climatic conditions: outdoors in summer, outdoors in spring, and indoors in a growth chamber. In particular, the regulation of stomatal conductance (g(s)), mesophyll conductance to CO(2) (g(m)), leaf photochemistry (chlorophyll fluorescence), and biochemistry (V(c,max)) were assessed. Leaf gas exchange and chlorophyll fluorescence data revealed that water stress induced a similar degree of stomatal closure and decreased A(N) under all three conditions, while V(c,max) was unaffected. However, the behaviour of g(m) differed depending on the climatic conditions. In outdoor plants, g(m) strongly declined with water stress, but it recovered rapidly (1-2 d) after re-watering in spring while it remained low many days after re-watering in summer. In indoor plants, g(m) initially declined with water stress, but then recovered to control values during the acclimation period. These differences were reflected in different velocities of recovery of A(N) after re-watering, being the slowest in outdoor summer plants and the fastest in indoor plants. It is suggested that these differences among the experiments are related to the prevailing climatic conditions, i.e. to the fact that stress factors other than water stress have been superimposed (e.g. excessive light and elevated temperature). In conclusion, besides g(s), g(m) contributes greatly to the limitation of photosynthesis during water stress and during

Quantification of plant cell coupling with three-dimensional photoactivation microscopy.

Science.gov (United States)

Liesche, J; Schulz, A

2012-07-01

Plant cells are directly connected by plasmodesmata that form channels through the cell wall and enable the intercellular movement of cytosolic solutes, membrane lipids and signalling molecules. Transport through plasmodesmata is regulated not only by a fixed size-exclusion limit, but also by physiological and pathological adaptation. To understand plant cell communication, carbon allocation and pathogen attack, the capacities for a specific molecule to pass a specific cell-wall interface is an essential parameter. So far, the degree of cell coupling was derived from frequency and diameter of plasmodesmata in relevant tissues as assessed by electron microscopy of fixed material. However, plasmodesmata functionality and capacity can only be determined in live material, not from electron microscopy, which is static and prone to fixation artefacts. Plasmodesmata functionality was a few times assessed using fluorescent tracers with diffusion properties similar to cytosolic solutes. Here, we used three-dimensional photoactivation microscopy to quantify plasmodesmata-mediated cell-wall permeability between living Cucurbita maxima leaf mesophyll cells with caged fluorescein as tracer. For the first time, all necessary functional and anatomical data were gathered for each individual cell from three-dimensional time series. This approach utilized a confocal microscope equipped with resonant scanner, which provides the high acquisition speed necessary to record optical sections of whole cells and offers time resolution high enough to follow the kinetics of photoactivation. The results were compared to two-dimensional measurements, which are shown to give a good estimate of cell coupling adequate for homogenous tissues. The two-dimensional approach is limited whenever tissues interfaces are studied that couple different cell types with diverse cell geometries. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

3D Plant Cell Architecture of Arabidopsis thaliana (Brassicaceae Using Focused Ion Beam–Scanning Electron Microscopy

Directory of Open Access Journals (Sweden)

Bhawana

2014-06-01

Full Text Available Premise of the study: Focused ion beam–scanning electron microscopy (FIB-SEM combines the ability to sequentially mill the sample surface and obtain SEM images that can be used to create 3D renderings with micron-level resolution. We have applied FIB-SEM to study Arabidopsis cell architecture. The goal was to determine the efficacy of this technique in plant tissue and cellular studies and to demonstrate its usefulness in studying cell and organelle architecture and distribution. Methods: Seed aleurone, leaf mesophyll, stem cortex, root cortex, and petal lamina from Arabidopsis were fixed and embedded for electron microscopy using protocols developed for animal tissues and modified for use with plant cells. Each sample was sectioned using the FIB and imaged with SEM. These serial images were assembled to produce 3D renderings of each cell type. Results: Organelles such as nuclei and chloroplasts were easily identifiable, and other structures such as endoplasmic reticula, lipid bodies, and starch grains were distinguishable in each tissue. Discussion: The application of FIB-SEM produced 3D renderings of five plant cell types and offered unique views of their shapes and internal content. These results demonstrate the usefulness of FIB-SEM for organelle distribution and cell architecture studies.

Xylogenesis in zinnia (Zinnia elegans) cell cultures: unravelling the regulatory steps in a complex developmental programmed cell death event.

Science.gov (United States)

Iakimova, Elena T; Woltering, Ernst J

2017-04-01

Physiological and molecular studies support the view that xylogenesis can largely be determined as a specific form of vacuolar programmed cell death (PCD). The studies in xylogenic zinnia cell culture have led to many breakthroughs in xylogenesis research and provided a background for investigations in other experimental models in vitro and in planta . This review discusses the most essential earlier and recent findings on the regulation of xylem elements differentiation and PCD in zinnia and other xylogenic systems. Xylogenesis (the formation of water conducting vascular tissue) is a paradigm of plant developmental PCD. The xylem vessels are composed of fused tracheary elements (TEs)-dead, hollow cells with patterned lignified secondary cell walls. They result from the differentiation of the procambium and cambium cells and undergo cell death to become functional post-mortem. The TE differentiation proceeds through a well-coordinated sequence of events in which differentiation and the programmed cellular demise are intimately connected. For years a classical experimental model for studies on xylogenesis was the xylogenic zinnia (Zinnia elegans) cell culture derived from leaf mesophyll cells that, upon induction by cytokinin and auxin, transdifferentiate into TEs. This cell system has been proven very efficient for investigations on the regulatory components of xylem differentiation which has led to many discoveries on the mechanisms of xylogenesis. The knowledge gained from this system has potentiated studies in other xylogenic cultures in vitro and in planta. The present review summarises the previous and latest findings on the hormonal and biochemical signalling, metabolic pathways and molecular and gene determinants underlying the regulation of xylem vessels differentiation in zinnia cell culture. Highlighted are breakthroughs achieved through the use of xylogenic systems from other species and newly introduced tools and analytical approaches to study the

Effects of actonomycin D and ultraviolet irradiation on multiplication of brome mosaic virus in host and non-host cells

International Nuclear Information System (INIS)

Maekawa, K.; Furusawa, I.; Okuno, T.

1981-01-01

The modes of multiplication of brome mosaic virus (BMV) were compared in protoplasts isolated from host and non-host plants. BMV actively multiplied in the leaves and isolated mesophyll protoplasts of barley, a host of BMV. BMV multiplication in barley protoplasts was inhibited by addition of actinomycin D immediately after inoculation or by u.v. irradiation of the protoplasts before inoculation. In contrast, although BMV could not multiply in leaves of radish and turnip (non-hosts for BMV) it multiplied at a low level in protoplasts isolated from these two plant species. Moreover, u.v. irradiation, or the addition of actinomycin D, enhanced multiplication of BMV in radish and turnip protoplasts. These results suggest that (i) in the host cells replication of BMV is dependent on cellular metabolism of nucleic acid and protein, and (ii) in the non-host cells a substance(s) inhibitory to replication of BMV is synthesized. (author)

Phloem unloading in developing leaves of sugar beet

International Nuclear Information System (INIS)

Schmalstig, J.G.

1985-01-01

Physiological and transport data support a symplastic pathway for phloem unloading in developing leaves of sugar beet (Beta vulgaris L. Klein E, multigerm). The sulfhydryl inhibitor parachloromercuribenzene sulfonic acid (PCMBS) inhibited uptake of [ 14 C]-sucrose added to the free space of developing leaves, but did not affect import of [ 14 C]-sucrose during steady-state 14 CO 2 labeling of a source leaf. The passively-transported xenobiotic sugar, [ 14 C]-L-glucose did not readily enter mesophyll cells when supplied through the cut end of the petiole of a sink leaf as determined by whole leaf autoradiography. In contrast, [ 14 C]-L-glucose translocated through the phloem from a mature leaf, rapidly entered mesophyll cells, and was evenly distributed between mesophyll and veins. Autoradiographs of developing leaves following a pulse of 14 CO 2 to a source leaf revealed rapid passage of phloem translocated into progressively higher order veins as the leaf developed. Entry into V order veins occurred during the last stage of import through the phloem. Import into developing leaves was inhibited by glyphosate (N-phosphomethylglycine), a herbicide which inhibits the aromatic amino acid pathway and hence protein synthesis. Glyphosate also stopped net starch accumulation in sprayed mature leaves, but did not affect export of carbon from treated leaves during the time period that import into developed leaves was inhibited

Non-linear direct effects of acid rain on leaf photosynthetic rate of terrestrial plants

NARCIS (Netherlands)

Dong, Dan; Du, Enzai; Sun, Zhengzhong; Zeng, Xuetong; Vries, de Wim

2017-01-01

Anthropogenic emissions of acid precursors have enhanced global occurrence of acid rain, especially in East Asia. Acid rain directly suppresses leaf function by eroding surface waxes and cuticle and leaching base cations from mesophyll cells, while the simultaneous foliar uptake of nitrates in

A 3-D Model of a Perennial Ryegrass Primary Cell Wall and Its Enzymatic Degradation

Directory of Open Access Journals (Sweden)

Indrakumar Vetharaniam

2014-05-01

Full Text Available We have developed a novel 3-D, agent-based model of cell-wall digestion to improve our understanding of ruminal cell-wall digestion. It offers a capability to study cell walls and their enzymatic modification, by providing a representation of cellulose microfibrils and non-cellulosic polysaccharides and by simulating their spatial and catalytic interactions with enzymes. One can vary cell-wall composition and the types and numbers of enzyme molecules, allowing the model to be applied to a range of systems where cell walls are degraded and to the modification of cell walls by endogenous enzymes. As a proof of principle, we have modelled the wall of a mesophyll cell from the leaf of perennial ryegrass and then simulated its enzymatic degradation. This is a primary, non-lignified cell wall and the model includes cellulose, hemicelluloses (glucuronoarabinoxylans, 1,3;1,4-β-glucans, and xyloglucans and pectin. These polymers are represented at the level of constituent monosaccharides, and assembled to form a 3-D, meso-scale representation of the molecular structure of the cell wall. The composition of the cell wall can be parameterised to represent different walls in different cell types and taxa. The model can contain arbitrary combinations of different enzymes. It simulates their random diffusion through the polymer networks taking collisions into account, allowing steric hindrance from cell-wall polymers to be modelled. Steric considerations are included when target bonds are encountered, and breakdown products resulting from enzymatic activity are predicted.

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Indian Academy of Sciences (India)

Madhu

ROS network is essential to induce disease resistance and even to mediate resistance to multiple stresses in ...... campestris and French bean mesophyll cells; Plant J. 15 .... He Y, Tang R H, Hao Y, Stevens R D, Cook C W, Ahn S M, Jing L,.

Anatomical studies of some medicinal plants of family polygonaceae

International Nuclear Information System (INIS)

Hameed, I.; Hussain, F.; Dastgir, G.

2010-01-01

Anatomical studies of the 6 different species of family Polygonaceae viz., Rumex hastatus D. Don, Rumex dentatus Linn, Rumex nepalensis Spreng, Rheum australe D. Don, Polygonum plebejum R. Br and Persicaria maculosa S.F. Gay are presented. The study is based on the presence and absence of epidermis, parenchyma, collenchyma, sclerenchyma, endodermis, pericycle, xylem, phloem, pith, mesophyll cells and stone cells. (author)

Photosynthesis-related characteristics of the midrib and the interveinal lamina in leaves of the C3-CAM intermediate plant Mesembryanthemum crystallinum.

Science.gov (United States)

Kuźniak, Elżbieta; Kornas, Andrzej; Kaźmierczak, Andrzej; Rozpądek, Piotr; Nosek, Michał; Kocurek, Maciej; Zellnig, Günther; Müller, Maria; Miszalski, Zbigniew

2016-06-01

Leaf veins are usually encircled by specialized bundle sheath cells. In C4 plants, they play an important role in CO2 assimilation, and the photosynthetic activity is compartmentalized between the mesophyll and the bundle sheath. In C3 and CAM (Crassulacean acid metabolism) plants, the photosynthetic activity is generally attributed to the leaf mesophyll cells, and the vascular parenchymal cells are rarely considered for their role in photosynthesis. Recent studies demonstrate that enzymes required for C4 photosynthesis are also active in the veins of C3 plants, and their vascular system contains photosynthetically competent parenchyma cells. However, our understanding of photosynthesis in veins of C3 and CAM plants still remains insufficient. Here spatial analysis of photosynthesis-related properties were applied to the midrib and the interveinal lamina cells in leaves of Mesembryanthemum crystallinum, a C3-CAM intermediate plant. The midrib anatomy as well as chloroplast structure and chlorophyll fluorescence, diurnal gas exchange profiles, the immunoblot patterns of PEPC (phosphoenolpyruvate carboxylase) and RubisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase), H2O2 localization and antioxidant enzyme activities were compared in the midrib and in the interveinal mesophyll cells in leaves of C3 and CAM plants. Leaf midribs were structurally competent to perform photosynthesis in C3 and CAM plants. The midrib chloroplasts resembled those in the bundle sheath cells of C4 plants and were characterized by limited photosynthetic activity. The metabolic roles of midrib chloroplasts differ in C3 and CAM plants. It is suggested that in leaves of C3 plants the midrib chloroplasts could be involved in the supply of CO2 for carboxylation, and in CAM plants they could provide malate to different metabolic processes and mediate H2O2 signalling. © The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For

On the contributions of photorespiration and compartmentation to the contrasting intramolecular 2H profiles of C3 and C4 plant sugars

Science.gov (United States)

Youping Zhou; Benli Zhang; Hilary Stuart-Williams; Kliti Grice; Charles H. Hocart; Arthur Gessler; Zachary E. Kayler; Graham D. Farquhar

2018-01-01

Compartmentation of C4 photosynthetic biochemistry into bundle sheath (BS) and mesophyll (M) cells, and photorespiration in C3 plants is predicted to have hydrogen isotopic consequences for metabolites at both molecular and site-specific levels. Molecular-level evidence was recently reported (Zhou et al., 2016), but...

Regulation of photosynthesis and stomatal and mesophyll conductance under water stress and recovery in olive trees: correlation with gene expression of carbonic anhydrase and aquaporins.

Science.gov (United States)

Perez-Martin, Alfonso; Michelazzo, Chiara; Torres-Ruiz, Jose M; Flexas, Jaume; Fernández, José E; Sebastiani, Luca; Diaz-Espejo, Antonio

2014-07-01

The hypothesis that aquaporins and carbonic anhydrase (CA) are involved in the regulation of stomatal (g s) and mesophyll (g m) conductance to CO2 was tested in a short-term water-stress and recovery experiment in 5-year-old olive plants (Olea europaea) growing outdoors. The evolution of leaf gas exchange, chlorophyll fluorescence, and plant water status, and a quantitative analysis of photosynthesis limitations, were followed during water stress and recovery. These variables were correlated with gene expression of the aquaporins OePIP1.1 and OePIP2.1, and stromal CA. At mild stress and at the beginning of the recovery period, stomatal limitations prevailed, while the decline in g m accounted for up to 60% of photosynthesis limitations under severe water stress. However, g m was restored to control values shortly after rewatering, facilitating the recovery of the photosynthetic rate. CA was downregulated during water stress and upregulated after recovery. The use of structural equation modelling allowed us to conclude that both OePIP1.1 and OePIP2.1 expression could explain most of the variations observed for g s and g m. CA expression also had a small but significant effect on g m in olive under water-stress conditions. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Elements Required for an Efficient NADP-Malic Enzyme Type C4 Photosynthesis1[C][W][OPEN

Science.gov (United States)

Wang, Yu; Long, Stephen P.; Zhu, Xin-Guang

2014-01-01

C4 photosynthesis has higher light, nitrogen, and water use efficiencies than C3 photosynthesis. Although the basic anatomical, cellular, and biochemical features of C4 photosynthesis are well understood, the quantitative significance of each element of C4 photosynthesis to the high photosynthetic efficiency are not well defined. Here, we addressed this question by developing and using a systems model of C4 photosynthesis, which includes not only the Calvin-Benson cycle, starch synthesis, sucrose synthesis, C4 shuttle, and CO2 leakage, but also photorespiration and metabolite transport between the bundle sheath cells and mesophyll cells. The model effectively simulated the CO2 uptake rates, and the changes of metabolite concentrations under varied CO2 and light levels. Analyses show that triose phosphate transport and CO2 leakage can help maintain a high photosynthetic rate by balancing ATP and NADPH amounts in bundle sheath cells and mesophyll cells. Finally, we used the model to define the optimal enzyme properties and a blueprint for C4 engineering. As such, this model provides a theoretical framework for guiding C4 engineering and studying C4 photosynthesis in general. PMID:24521879

Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells

KAUST Repository

Hu, Honghong

2009-12-13

The continuing rise in atmospheric CO2 causes stomatal pores in leaves to close and thus globally affects CO2 influx into plants, water use efficiency and leaf heat stress. However, the CO2-binding proteins that control this response remain unknown. Moreover, which cell type responds to CO2, mesophyll or guard cells, and whether photosynthesis mediates this response are matters of debate. We demonstrate that Arabidopsis thaliana double-mutant plants in the beta-carbonic anhydrases betaCA1 and betaCA4 show impaired CO2-regulation of stomatal movements and increased stomatal density, but retain functional abscisic-acid and blue-light responses. betaCA-mediated CO2-triggered stomatal movements are not, in first-order, linked to whole leaf photosynthesis and can function in guard cells. Furthermore, guard cell betaca-overexpressing plants exhibit instantaneous enhanced water use efficiency. Guard cell expression of mammalian alphaCAII complements the reduced sensitivity of ca1 ca4 plants, showing that carbonic anhydrase-mediated catalysis is an important mechanism for betaCA-mediated CO2-induced stomatal closure and patch clamp analyses indicate that CO2/HCO3- transfers the signal to anion channel regulation. These findings, together with ht1-2 (ref. 9) epistasis analysis demonstrate that carbonic anhydrases function early in the CO2 signalling pathway, which controls gas-exchange between plants and the atmosphere.

Let’s not forget the critical role of surface tension in xylem water relations

Science.gov (United States)

Jean-Christophe Domec

2011-01-01

The widely supported cohesion–tension theory of water transport explains the importance of a continuous water column and the mechanism of long-distance ascent of sap in plants (Dixon 1914, Tyree 2003, Angeles et al. 2004). The evaporation of water from the surfaces of mesophyll cells causes the air–water interface to retreat into the cellulose matrix of the plant cell...

Diffusion and bulk flow in phloem loading

DEFF Research Database (Denmark)

Dölger, Julia; Rademaker, Hanna; Liesche, Johannes

2014-01-01

Plants create sugar in the mesophyll cells of their leaves by photosynthesis. This sugar, mostly sucrose, has to be loaded via the bundle sheath into the phloem vascular system (the sieve elements), where it is distributed to growing parts of the plant. We analyze the feasibility of a particular...

Interactive Influence of N and P on their uptake by four different ...

African Journals Online (AJOL)

The uptake kinetics of nitrogen (N) and phosphorus (P) by hydrophytes can be influenced by the interaction between N and P. In this study, Pistia stratiotes (a floating plant), Eichhornia crassipes (a floating plant), Vallisneria spiralis (a submerged plant), and Cyperus papyrus (an emergent plant) were selected to measure ...

Reproductive Ecology of Vallisneria americana Michaux

Science.gov (United States)

2006-12-01

River and Estuary, Maryland and Virginia, May 1978 to November 1981. U.S. Geologi- cal Survey Water Supply Paper 2234A. Carter, V. A., and N. B...Information Network, reference website: http://www.ars-grin.gov/. Haccius, B. 1952. Über die Blattstellung einiger Hydrocharitaceen-Embryonen. Planta 40: 333...Hydrocharitaceae). Aquatic Botany 13: 269- 298. Marie-Victorin, F. 1943. Les Vallisnéries américaines. Contributions de l’Institut Botanique de l’Universite de

Pectinous cell wall thickenings formation - A common defense strategy of plants to cope with Pb.

Science.gov (United States)

Krzesłowska, Magdalena; Rabęda, Irena; Basińska, Aneta; Lewandowski, Michał; Mellerowicz, Ewa J; Napieralska, Anna; Samardakiewicz, Sławomir; Woźny, Adam

2016-07-01

Lead, one of the most abundant and hazardous trace metals affecting living organisms, has been commonly detected in plant cell walls including some tolerant plants, mining ecotypes and hyperaccumulators. We have previously shown that in tip growing Funaria sp. protonemata cell wall is remodeled in response to lead by formation of thickenings rich in low-methylesterified pectins (pectin epitope JIM5 - JIM5-P) able to bind metal ions, which accumulate large amounts of Pb. Hence, it leads to the increase of cell wall capacity for Pb compartmentalization. Here we show that diverse plant species belonging to different phyla (Arabidopsis, hybrid aspen, star duckweed), form similar cell wall thickenings in response to Pb. These thickenings are formed in tip growing cells such as the root hairs, and in diffuse growing cells such as meristematic and root cap columella cells of root apices in hybrid aspen and Arabidopsis and in mesophyll cells in star duckweed fronds. Notably, all analyzed cell wall thickenings were abundant in JIM5-P and accumulated high amounts of Pb. In addition, the co-localization of JIM5-P and Pb commonly occurred in these cells. Hence, cell wall thickenings formed the extra compartment for Pb accumulation. In this way plant cells increased cell wall capacity for compartmentalization of this toxic metal, protecting protoplast from its toxicity. As cell wall thickenings occurred in diverse plant species and cell types differing in the type of growth we may conclude that pectinous cell wall thickenings formation is a widespread defense strategy of plants to cope with Pb. Moreover, detection of natural defense strategy, increasing plant cell walls capacity for metal accumulation, reveals a promising direction for enhancing plant efficiency in phytoremediation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Leaf responses to drought stress in Mediterranean accessions of Solanum lycopersicum: anatomical adaptations in relation to gas exchange parameters.

Science.gov (United States)

Galmés, Jeroni; Ochogavía, Joan Manuel; Gago, Jorge; Roldán, Emilio José; Cifre, Josep; Conesa, Miquel Àngel

2013-05-01

In a previous study, important acclimation to water stress was observed in the Ramellet tomato cultivar (TR) from the Balearic Islands, related to an increase in the water-use efficiency through modifications in both stomatal (g(s)) and mesophyll conductances (g(m)). In the present work, the comparison of physiological and morphological traits between TR accessions grown with and without water stress confirmed that variability in the photosynthetic capacity was mostly explained by differences in the diffusion of CO2 through stomata and leaf mesophyll. Maximization of gm under both treatments was mainly achieved through adjustments in the mesophyll thickness and porosity and the surface area of chloroplasts exposed to intercellular airspace (S(c)). In addition, the lower g(m) /S(c) ratio for a given porosity in drought-acclimated plants suggests that the decrease in gm was due to an increased cell wall thickness. Stomatal conductance was also affected by drought-associated changes in the morphological properties of stomata, in an accession and treatment-dependent manner. The results confirm the presence of advantageous physiological traits in the response to drought stress in Mediterranean accessions of tomato, and relate them to particular changes in the leaf anatomical properties, suggesting specific adaptive processes operating at the leaf anatomical level. © 2012 Blackwell Publishing Ltd.

Abscisic Acid–Responsive Guard Cell Metabolomes of Arabidopsis Wild-Type and gpa1 G-Protein Mutants[C][W

Science.gov (United States)

Jin, Xiaofen; Wang, Rui-Sheng; Zhu, Mengmeng; Jeon, Byeong Wook; Albert, Reka; Chen, Sixue; Assmann, Sarah M.

2013-01-01

Individual metabolites have been implicated in abscisic acid (ABA) signaling in guard cells, but a metabolite profile of this specialized cell type is lacking. We used liquid chromatography–multiple reaction monitoring mass spectrometry for targeted analysis of 85 signaling-related metabolites in Arabidopsis thaliana guard cell protoplasts over a time course of ABA treatment. The analysis utilized ∼350 million guard cell protoplasts from ∼30,000 plants of the Arabidopsis Columbia accession (Col) wild type and the heterotrimeric G-protein α subunit mutant, gpa1, which has ABA-hyposensitive stomata. These metabolomes revealed coordinated regulation of signaling metabolites in unrelated biochemical pathways. Metabolites clustered into different temporal modules in Col versus gpa1, with fewer metabolites showing ABA-altered profiles in gpa1. Ca2+-mobilizing agents sphingosine-1-phosphate and cyclic adenosine diphosphate ribose exhibited weaker ABA-stimulated increases in gpa1. Hormone metabolites were responsive to ABA, with generally greater responsiveness in Col than in gpa1. Most hormones also showed different ABA responses in guard cell versus mesophyll cell metabolomes. These findings suggest that ABA functions upstream to regulate other hormones, and are also consistent with G proteins modulating multiple hormonal signaling pathways. In particular, indole-3-acetic acid levels declined after ABA treatment in Col but not gpa1 guard cells. Consistent with this observation, the auxin antagonist α-(phenyl ethyl-2-one)-indole-3-acetic acid enhanced ABA-regulated stomatal movement and restored partial ABA sensitivity to gpa1. PMID:24368793

Biology of Leptoypha hospita (Hemiptera: Tingidae), a Potential Biological Control Agent of Chinese Privet

Science.gov (United States)

Yanzhuo Zhang; James L. Hanula; Scott Horn; Kristine Braman; Jianghua Sun

2011-01-01

The biology of Leptoypha hospita Drake et Poor (Hemiptera: Tingidae), a potential biological control agent from China for Chinese privet, Ligustrum sinense Lour., was studied in quarantine in the United States. Both nymphs and adults feed on Chinese privet mesophyll cells that lead to a bleached appearance of leaves and dieback of branch tips. L. hospita has five...

Pengaruh penambahan enzim dan waktu inkubasi terhadap jumlah protoplas mesofil daun anggrek Dendrobium Sp

Directory of Open Access Journals (Sweden)

Edi Setiti Wida Utami

1995-12-01

Full Text Available This research was done in order to study of effect the supplementation of enzyme and incubation time for the protoplast qualtity leaf mesophyll Dendrobium sp. Protoplast can be used for culture protoplast, for somatic cross, biology research, and material for genetic manipulation. This research to make use of material was Dendrobium sp Orchid. Explant that to used was leaf mesophyll Dendrobium sp. For the protoplast isolation to used enzyme combination selulase and maserozym (Onozuka R-10. Yakult JONSHA Co., Ltd., (Japan_ with concentration selulase : maserozym is (0.5; 0.05; 0.75; 0.075; 0; 0.1. leaf mesophyll Dendrobium sp. to get a soak in enzyme solution with incubation time 12-13 hours and 14-1 hours.the result shown that enzyme combination selulase and maserozym can be used for protoplast isolation leaf mesophyll Dendrobium sp Orchid. The best enzyme concertration to fit for isolatin protoplast leaf mesophyll Dendrobium sp to be selulase 0.75% and maserozym 0.075% with incubation time 14-15 hours.

Persistent negative temperature response of mesophyll conductance in red raspberry (Rubus idaeus L.) leaves under both high and low vapour pressure deficits: a role for abscisic acid?

Science.gov (United States)

Qiu, Changpeng; Ethier, Gilbert; Pepin, Steeve; Dubé, Pascal; Desjardins, Yves; Gosselin, André

2017-09-01

The temperature dependence of mesophyll conductance (g m ) was measured in well-watered red raspberry (Rubus idaeus L.) plants acclimated to leaf-to-air vapour pressure deficit (VPDL) daytime differentials of contrasting amplitude, keeping a fixed diurnal leaf temperature (T leaf ) rise from 20 to 35 °C. Contrary to the great majority of g m temperature responses published to date, we found a pronounced reduction of g m with increasing T leaf irrespective of leaf chamber O 2 level and diurnal VPDL regime. Leaf hydraulic conductance was greatly enhanced during the warmer afternoon periods under both low (0.75 to 1.5 kPa) and high (0.75 to 3.5 kPa) diurnal VPDL regimes, unlike stomatal conductance (g s ), which decreased in the afternoon. Consequently, the leaf water status remained largely isohydric throughout the day, and therefore cannot be evoked to explain the diurnal decrease of g m . However, the concerted diurnal reductions of g m and g s were well correlated with increases in leaf abscisic acid (ABA) content, thus suggesting that ABA can induce a significant depression of g m under favourable leaf water status. Our results challenge the view that the temperature dependence of g m can be explained solely from dynamic leaf anatomical adjustments and/or from the known thermodynamic properties of aqueous solutions and lipid membranes.​. © 2017 John Wiley & Sons Ltd.

Long-distance signaling within Coleus x hybridus leaves; mediated by changes in intra-leaf CO2?

Science.gov (United States)

Stahlberg, R.; Van Volkenburgh, E.; Cleland, R. E.

2001-01-01

Rapid long-distance signaling in plants can occur via several mechanisms, including symplastic electric coupling and pressure waves. We show here in variegated Coleus leaves a rapid propagation of electrical signals that appears to be caused by changes in intra-leaf CO2 concentrations. Green leaf cells, when illuminated, undergo a rapid depolarization of their membrane potential (Vm) and an increase in their apoplastic pH (pHa) by a process that requires photosynthesis. This is followed by a slower hyperpolarization of Vm and apoplastic acidification, which do not require photosynthesis. White (chlorophyll-lacking) leaf cells, when in isolated white leaf segments, show only the slow response, but when in mixed (i.e. green and white) segments, the rapid Vm depolarization and increase in pHa propagate over more than 10 mm from the green to the white cells. Similarly, these responses propagate 12-20 mm from illuminated to unilluminated green cells. The fact that the propagation of these responses is eliminated when the leaf air spaces are infiltrated with solution indicates that the signal moves in the apoplast rather than the symplast. A depolarization of the mesophyll cells is induced in the dark by a decrease in apoplastic CO2 but not by an increase in pHa. These results support the hypothesis that the propagating signal for the depolarization of the white mesophyll cells is a photosynthetically induced decrease in the CO2 level of the air spaces throughout the leaf.

Regulators of floral fragrance production and their target genes in petunia are not exclusively active in the epidermal cells of petals.

Science.gov (United States)

Van Moerkercke, Alex; Galván-Ampudia, Carlos S; Verdonk, Julian C; Haring, Michel A; Schuurink, Robert C

2012-05-01

In which cells of the flower volatile biosynthesis takes place is unclear. In rose and snapdragon, some enzymes of the volatile phenylpropanoid/benzenoid pathway have been shown to be present in the epidermal cells of petals. It is therefore generally believed that the production of these compounds occurs in these cells. However, whether the entire pathway is active in these cells and whether it is exclusively active in these cells remains to be proven. Cell-specific transcription factors activating these genes will determine in which cells they are expressed. In petunia, the transcription factor EMISSION OF BENZENOIDS II (EOBII) activates the ODORANT1 (ODO1) promoter and the promoter of the biosynthetic gene isoeugenol synthase (IGS). The regulator ODO1 in turn activates the promoter of the shikimate gene 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Here the identification of a new target gene of ODO1, encoding an ABC transporter localized on the plasma membrane, PhABCG1, which is co-expressed with ODO1, is described. PhABCG1 expression is up-regulated in petals overexpressing ODO1 through activation of the PhABCG1 promoter. Interestingly, the ODO1, PhABCG1, and IGS promoters were active in petunia protoplasts originating from both epidermal and mesophyll cell layers of the petal, suggesting that the volatile phenylpropanoid/benzenoid pathway in petunia is active in these different cell types. Since volatile release occurs from epidermal cells, trafficking of (volatile) compounds between cell layers must be involved, but the exact function of PhABCG1 remains to be resolved.

The Arabidopsis vacuolar malate channel is a member of the ALMT family.

Science.gov (United States)

Kovermann, Peter; Meyer, Stefan; Hörtensteiner, Stefan; Picco, Cristiana; Scholz-Starke, Joachim; Ravera, Silvia; Lee, Youngsook; Martinoia, Enrico

2007-12-01

In plants, malate is a central metabolite and fulfills a large number of functions. Vacuolar malate may reach very high concentrations and fluctuate rapidly, whereas cytosolic malate is kept at a constant level allowing optimal metabolism. Recently, a vacuolar malate transporter (Arabidopsis thaliana tonoplast dicarboxylate transporter, AttDT) was identified that did not correspond to the well-characterized vacuolar malate channel. We therefore hypothesized that a member of the aluminum-activated malate transporter (ALMT) gene family could code for a vacuolar malate channel. Using GFP fusion constructs, we could show that AtALMT9 (A. thaliana ALMT9) is targeted to the vacuole. Promoter-GUS fusion constructs demonstrated that this gene is expressed in all organs, but is cell-type specific as GUS activity in leaves was detected nearly exclusively in mesophyll cells. Patch-clamp analysis of an Atalmt9 T-DNA insertion mutant exhibited strongly reduced vacuolar malate channel activity. In order to functionally characterize AtALMT9 as a malate channel, we heterologously expressed this gene in tobacco and in oocytes. Overexpression of AtALMT9-GFP in Nicotiana benthamiana leaves strongly enhanced the malate current densities across the mesophyll tonoplasts. Functional expression of AtALMT9 in Xenopus oocytes induced anion currents, which were clearly distinguishable from endogenous oocyte currents. Our results demonstrate that AtALMT9 is a vacuolar malate channel. Deletion mutants for AtALMT9 exhibit only slightly reduced malate content in mesophyll protoplasts and no visible phenotype, indicating that AttDT and the residual malate channel activity are sufficient to sustain the transport activity necessary to regulate the cytosolic malate homeostasis.

Mixing of maize and wheat genomic DNA by somatic hybridization in regenerated sterile maize plants.

Science.gov (United States)

Szarka, B.; Göntér, I.; Molnár-Láng, M.; Mórocz, S.; Dudits, D.

2002-07-01

Intergeneric somatic hybridization was performed between albino maize ( Zea mays L.) protoplasts and mesophyll protoplasts of wheat ( Triticum aestivum L.) by polyethylene glycol (PEG) treatments. None of the parental protoplasts were able to produce green plants without fusion. The maize cells regenerated only rudimentary albino plantlets of limited viability, and the wheat mesophyll protoplasts were unable to divide. PEG-mediated fusion treatments resulted in hybrid cells with mixed cytoplasm. Six months after fusion green embryogenic calli were selected as putative hybrids. The first-regenerates were discovered as aborted embryos. Regeneration of intact, green, maize-like plants needed 6 months of further subcultures on hormone-free medium. These plants were sterile, although had both male and female flowers. The cytological analysis of cells from callus tissues and root tips revealed 56 chromosomes, but intact wheat chromosomes were not observed. Using total DNA from hybrid plants, three RAPD primer combinations produced bands resembling the wheat profile. Genomic in situ hybridization (GISH) using total wheat DNA as a probe revealed the presence of wheat DNA islands in the maize chromosomal background. The increased viability and the restored green color were the most-significant new traits as compared to the original maize parent. Other intermediate morphological traits of plants with hybrid origin were not found.

Directly Transforming PCR-Amplified DNA Fragments into Plant Cells Is a Versatile System That Facilitates the Transient Expression Assay

Science.gov (United States)

Lu, Yuming; Chen, Xi; Wu, Yuxuan; Wang, Yanping; He, Yuqing; Wu, Yan

2013-01-01

A circular plasmid containing a gene coding sequence has been broadly used for studying gene regulation in cells. However, to accommodate a quick screen plasmid construction and preparation can be time consuming. Here we report a PCR amplified dsDNA fragments (PCR-fragments) based transient expression system (PCR-TES) for suiting in the study of gene regulation in plant cells. Instead of transforming plasmids into plant cells, transient expression of PCR-fragments can be applicable. The transformation efficiency and expression property of PCR-fragments are comparable to transformation using plasmids. We analyzed the transformation efficiency in PCR-TES at transcription and protein levels. Our results indicate that the PCR-TES is as versatile as the conventional transformation system using plasmid DNA. Through reconstituting PYR1-mediated ABA signaling pathway in Arabidopsis mesophyll protoplasts, we were not only validating the practicality of PCR-TES but also screening potential candidates of CDPK family members which might be involved in the ABA signaling. Moreover, we determined that phosphorylation of ABF2 by CPK4 could be mediated by ABA-induced PYR1 and ABI1, demonstrating a crucial role of CDPKs in the ABA signaling. In summary, PCR-TES can be applicable to facilitate analyzing gene regulation and for the screen of putative regulatory molecules at the high throughput level in plant cells. PMID:23468926

Directly transforming PCR-amplified DNA fragments into plant cells is a versatile system that facilitates the transient expression assay.

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Yuming Lu

Full Text Available A circular plasmid containing a gene coding sequence has been broadly used for studying gene regulation in cells. However, to accommodate a quick screen plasmid construction and preparation can be time consuming. Here we report a PCR amplified dsDNA fragments (PCR-fragments based transient expression system (PCR-TES for suiting in the study of gene regulation in plant cells. Instead of transforming plasmids into plant cells, transient expression of PCR-fragments can be applicable. The transformation efficiency and expression property of PCR-fragments are comparable to transformation using plasmids. We analyzed the transformation efficiency in PCR-TES at transcription and protein levels. Our results indicate that the PCR-TES is as versatile as the conventional transformation system using plasmid DNA. Through reconstituting PYR1-mediated ABA signaling pathway in Arabidopsis mesophyll protoplasts, we were not only validating the practicality of PCR-TES but also screening potential candidates of CDPK family members which might be involved in the ABA signaling. Moreover, we determined that phosphorylation of ABF2 by CPK4 could be mediated by ABA-induced PYR1 and ABI1, demonstrating a crucial role of CDPKs in the ABA signaling. In summary, PCR-TES can be applicable to facilitate analyzing gene regulation and for the screen of putative regulatory molecules at the high throughput level in plant cells.

Synthesis of viral DNA forms in Nicotiana plumbaginifolia protoplasts inoculated with cassava latent virus (CLV); evidence for the independent replication of one component of the CLV genome.

OpenAIRE

Townsend, R; Watts, J; Stanley, J

1986-01-01

Totipotent leaf mesophyll protoplasts of Nicotiana plumbaginifolia, Viviani were inoculated with cassava latent virus (CLV) or with full length copies of CLV genomic DNAs 1 and 2 excised from replicative forms of M13 clones. Virus specific DNAs began to appear 48-72h after inoculation with virus or cloned DNAs, coincident with the onset of host cell division. Infected cells accumulated supercoiled forms of DNAs 1 and 2 as well as progeny single-stranded (ss) virion (+) sense DNAs representing...

Cadmium induced changes in subcellular glutathione contents within glandular trichomes of Cucurbita pepo L.

Science.gov (United States)

Kolb, Dagmar; Müller, Maria; Zellnig, Günther; Zechmann, Bernd

2010-07-01

Plants cope with cadmium (Cd) stress by complexation with phytochelatins (Pc), metallothioneins and glutathione and sequestration within vacuoles. Especially glutathione was found to play a major role in Cd detoxification as Cd shows a high binding affinity towards thiols and as glutathione is a precursor for Pc synthesis. In the present study, we have used an immunohistochemical approach combined with computer-supported transmission electron microscopy in order to measure changes in the subcellular distribution of glutathione during Cd-stress in mesophyll cells and cells of different glandular trichomes (long and short stalked) of Cucurbita pepo L. subsp. pepo var. styriaca GREB: . Even though no ultrastructural alterations were observed in leaf and glandular trichome cells after the treatment of plants with 50 microM cadmium chloride (CdCl(2)) for 48 h, all cells showed a large decrease in glutathione contents. The strongest decrease was found in nuclei and the cytosol (up to 76%) in glandular trichomes which are considered as a major side of Cd accumulation in leaves. The ratio of glutathione between the cytosol and nuclei and the other cell compartments was strongly decreased only in glandular trichomes (more than 50%) indicating that glutathione in these two cell compartments is especially important for the detoxification of Cd in glandular trichomes. Additionally, these data indicate that large amounts of Cd are withdrawn from nuclei during Cd exposure. The present study gives a detailed insight into the compartment-specific importance of glutathione during Cd exposure in mesophyll cells and glandular trichomes of C. pepo L. plants.

Integrated Multi-Trophic Recirculating Aquaculture System for Nile Tilapia (Oreochlomis niloticus

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Puchong Sri-uam

2016-06-01

Full Text Available Three densities of the sex-reversed male Nile tilapia, Oreochromis niloticus (20, 25, 50 fish/m3 were cultivated in an integrated multi-trophic recirculating aquaculture system (IMRAS that involves the ecological relationship between several living organisms, i.e., phytoplankton, zooplankton, and aquatic plants. The results indicated that, by providing proper interdependency between various species of living organisms, the concentrations of ammonia, nitrite, nitrate, and phosphate in the system were maintained below dangerous levels for Nile tilapia throughout the cultivation period. The highest wet weight productivity of Nile tilapia of 11 ± 1 kg was achieved at a fish density of 50 fish/m3. The aquatic plants in the treatment tank could effectively uptake the unwanted nitrogen (N and phosphorus (P compounds with the highest removal efficiencies of 9.52% and 11.4%, respectively. The uptake rates of nitrogen and phosphorus by aquatic plants could be ranked from high to low as: Egeria densa > Ceratophyllum demersum > Vallisneria spiralis and Vallisneria americana > Hygrophila difformis. The remaining N was further degraded through nitrification process, whereas the remaining P could well precipitate in the soil sediment in the treatment tank.

Submerged macrophytes modify bacterial community composition in sediments in a large, shallow, freshwater lake.

Science.gov (United States)

Zhao, Da-Yong; Liu, Peng; Fang, Chao; Sun, Yi-Meng; Zeng, Jin; Wang, Jian-Qun; Ma, Ting; Xiao, Yi-Hong; Wu, Qinglong L

2013-04-01

Submerged aquatic macrophytes are an important part of the lacustrine ecosystem. In this study, the bacterial community compositions in the rhizosphere sediments from three kinds of submerged macrophytes (Ceratophyllum demersum, Potamogeton crispus, and Vallisneria natans) were investigated to determine whether submerged macrophytes could drive the variation of bacterial community in the eutrophic Taihu Lake, China. Molecular techniques, including terminal restriction fragment length polymorphism (T-RFLP) of PCR-amplified 16S rRNA gene and clone libraries, were employed to analyze the bacterial community compositions. Remarkable differences of the T-RFLP patterns were observed among the different samples, and the results of LIBSHUFF analysis also confirmed that the bacterial community compositions in the rhizosphere sediments of three kinds of submerged macrophytes were statistically different from that of the unvegetated sediment. Acidobacteria, Deltaproteobacteria, and Betaproteobacteria were the dominant bacterial groups in the rhizosphere sediments of Ceratophyllum demersum, Potamogeton crispus, and Vallisneria natans, respectively, accounting for 15.38%, 29.03%, and 18.00% of the total bacterial abundances. Our study demonstrated that submerged macrophytes could influence the bacterial community compositions in their rhizosphere sediments, suggesting that macrophytes have an effect on the cycling and transportation of nutrients in the freshwater lake ecosystem.

Effects of snails, submerged plants and their coexistence on eutrophication in aquatic ecosystems

OpenAIRE

Mo Shuqing; Zhang Xiufeng; Tang Yali; Liu Zhengwen; Kettridge Nicholas

2017-01-01

Eutrophication resulting from nutrient loading to freshwater habitats is a severe problem, leading to degradation of ecosystems, including deterioration of water quality, water clarity and loss of biodiversity. Measures enacted to restore degraded freshwater ecosystems often involve the reintroduction of submerged plants and aquatic animals with beneficial ecological functions. In a mesocosm experiment, three treatments (planting with Vallisneria natans, introduction of the snail Bellamya aer...

Combining laser-assisted microdissection (LAM) and RNA-seq allows to perform a comprehensive transcriptomic analysis of epidermal cells of Arabidopsis embryo.

Science.gov (United States)

Sakai, Kaori; Taconnat, Ludivine; Borrega, Nero; Yansouni, Jennifer; Brunaud, Véronique; Paysant-Le Roux, Christine; Delannoy, Etienne; Martin Magniette, Marie-Laure; Lepiniec, Loïc; Faure, Jean Denis; Balzergue, Sandrine; Dubreucq, Bertrand

2018-01-01

Genome-wide characterization of tissue- or cell-specific gene expression is a recurrent bottleneck in biology. We have developed a sensitive approach based on ultra-low RNA sequencing coupled to laser assisted microdissection for analyzing different tissues of the small Arabidopsis embryo. We first characterized the number of genes detected according to the quantity of tissue yield and total RNA extracted. Our results revealed that as low as 0.02 mm 2 of tissue and 50 pg of total RNA can be used without compromising the number of genes detected. The optimised protocol was used to compare the epidermal versus mesophyll cell transcriptomes of cotyledons at the torpedo-shaped stage of embryo development. The approach was validated by the recovery of well-known epidermal genes such AtML1 or AtPDF2 and genes involved in flavonoid and cuticular waxes pathways. Moreover, the interest and sensitivity of this approach were highlighted by the characterization of several transcription factors preferentially expressed in epidermal cells. This technical advance unlocks some current limitations of transcriptomic analyses and allows to investigate further and efficiently new biological questions for which only a very small amounts of cells need to be isolated. For instance, it paves the way to increasing the spatial accuracy of regulatory networks in developing small embryo of Arabidopsis or other plant tissues.

Ecological anatomy of ferns fronds

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Nina M. Derzhavina

2014-04-01

Full Text Available Structural types of frond anatomy are distinguished on the basis of investigation of 30 species of homosporous ferns and with regard for literature: hydromorphic, hygromorphic, mesomorphic, subxeromorphic, and subsucculent (cryptic succulent. Following frond traits are of highest adaptive value: their area and thickness, type of mesophyll, dry weight of an area unit – specific superficial density, cellular volume, and number of cells per unit of frond area.

Isolation of a strong Arabidopsis guard cell promoter and its potential as a research tool

Science.gov (United States)

Yang, Yingzhen; Costa, Alex; Leonhardt, Nathalie; Siegel, Robert S; Schroeder, Julian I

2008-01-01

applicable to reduce specific gene expression in guard cells, providing a method for circumvention of limitations arising from genetic redundancy and lethality. These advances could be very useful for manipulating signaling pathways in guard cells and modifying plant performance under stress conditions. In addition, new guard cell and mesophyll cell-specific 23,000 gene microarray data are made publicly available here. PMID:18284694

Limited acclimation in leaf anatomy to experimental drought in tropical rainforest trees.

Science.gov (United States)

Binks, Oliver; Meir, Patrick; Rowland, Lucy; da Costa, Antonio Carlos Lola; Vasconcelos, Steel Silva; de Oliveira, Alex Antonio Ribeiro; Ferreira, Leandro; Mencuccini, Maurizio

2016-12-01

Dry periods are predicted to become more frequent and severe in the future in some parts of the tropics, including Amazonia, potentially causing reduced productivity, higher tree mortality and increased emissions of stored carbon. Using a long-term (12 year) through-fall exclusion (TFE) experiment in the tropics, we test the hypothesis that trees produce leaves adapted to cope with higher levels of water stress, by examining the following leaf characteristics: area, thickness, leaf mass per area, vein density, stomatal density, the thickness of palisade mesophyll, spongy mesophyll and both of the epidermal layers, internal cavity volume and the average cell sizes of the palisade and spongy mesophyll. We also test whether differences in leaf anatomy are consistent with observed differential drought-induced mortality responses among taxa, and look for relationships between leaf anatomy, and leaf water relations and gas exchange parameters. Our data show that trees do not produce leaves that are more xeromorphic in response to 12 years of soil moisture deficit. However, the drought treatment did result in increases in the thickness of the adaxial epidermis (TFE: 20.5 ± 1.5 µm, control: 16.7 ± 1.0 µm) and the internal cavity volume (TFE: 2.43 ± 0.50 mm 3 cm -2 , control: 1.77 ± 0.30 mm 3 cm -2 ). No consistent differences were detected between drought-resistant and drought-sensitive taxa, although interactions occurred between drought-sensitivity status and drought treatment for the palisade mesophyll thickness (P = 0.034) and the cavity volume of the leaves (P = 0.025). The limited response to water deficit probably reflects a tight co-ordination between leaf morphology, water relations and photosynthetic properties. This suggests that there is little plasticity in these aspects of plant anatomy in these taxa, and that phenotypic plasticity in leaf traits may not facilitate the acclimation of Amazonian trees to the predicted future reductions in dry

Pectin Methylesterification Impacts the Relationship between Photosynthesis and Plant Growth1[OPEN

Science.gov (United States)

Kim, Sang-Jin; Renna, Luciana; Brandizzi, Federica

2016-01-01

Photosynthesis occurs in mesophyll cells of specialized organs such as leaves. The rigid cell wall encapsulating photosynthetic cells controls the expansion and distribution of cells within photosynthetic tissues. The relationship between photosynthesis and plant growth is affected by leaf area. However, the underlying genetic mechanisms affecting carbon partitioning to different aspects of leaf growth are not known. To fill this gap, we analyzed Arabidopsis plants with altered levels of pectin methylesterification, which is known to modulate cell wall plasticity and plant growth. Pectin methylesterification levels were varied through manipulation of cotton Golgi-related (CGR) 2 or 3 genes encoding two functionally redundant pectin methyltransferases. Increased levels of methylesterification in a line over-expressing CGR2 (CGR2OX) resulted in highly expanded leaves with enhanced intercellular air spaces; reduced methylesterification in a mutant lacking both CGR-genes 2 and 3 (cgr2/3) resulted in thin but dense leaf mesophyll that limited CO2 diffusion to chloroplasts. Leaf, root, and plant dry weight were enhanced in CGR2OX but decreased in cgr2/3. Differences in growth between wild type and the CGR-mutants can be explained by carbon partitioning but not by variations in area-based photosynthesis. Therefore, photosynthesis drives growth through alterations in carbon partitioning to new leaf area growth and leaf mass per unit leaf area; however, CGR-mediated pectin methylesterification acts as a primary factor in this relationship through modulation of the expansion and positioning of the cells in leaves, which in turn drive carbon partitioning by generating dynamic carbon demands in leaf area growth and leaf mass per unit leaf area. PMID:27208234

Pectin Methylesterification Impacts the Relationship between Photosynthesis and Plant Growth.

Science.gov (United States)

M Weraduwage, Sarathi; Kim, Sang-Jin; Renna, Luciana; C Anozie, Fransisca; D Sharkey, Thomas; Brandizzi, Federica

2016-06-01

Photosynthesis occurs in mesophyll cells of specialized organs such as leaves. The rigid cell wall encapsulating photosynthetic cells controls the expansion and distribution of cells within photosynthetic tissues. The relationship between photosynthesis and plant growth is affected by leaf area. However, the underlying genetic mechanisms affecting carbon partitioning to different aspects of leaf growth are not known. To fill this gap, we analyzed Arabidopsis plants with altered levels of pectin methylesterification, which is known to modulate cell wall plasticity and plant growth. Pectin methylesterification levels were varied through manipulation of cotton Golgi-related (CGR) 2 or 3 genes encoding two functionally redundant pectin methyltransferases. Increased levels of methylesterification in a line over-expressing CGR2 (CGR2OX) resulted in highly expanded leaves with enhanced intercellular air spaces; reduced methylesterification in a mutant lacking both CGR-genes 2 and 3 (cgr2/3) resulted in thin but dense leaf mesophyll that limited CO2 diffusion to chloroplasts. Leaf, root, and plant dry weight were enhanced in CGR2OX but decreased in cgr2/3. Differences in growth between wild type and the CGR-mutants can be explained by carbon partitioning but not by variations in area-based photosynthesis. Therefore, photosynthesis drives growth through alterations in carbon partitioning to new leaf area growth and leaf mass per unit leaf area; however, CGR-mediated pectin methylesterification acts as a primary factor in this relationship through modulation of the expansion and positioning of the cells in leaves, which in turn drive carbon partitioning by generating dynamic carbon demands in leaf area growth and leaf mass per unit leaf area. © 2016 American Society of Plant Biologists. All Rights Reserved.

Altered physiology, cell structure, and gene expression of Theobroma cacao seedlings subjected to Cu toxicity.

Science.gov (United States)

Souza, Vânia L; de Almeida, Alex-Alan F; Souza, Jadiel de S; Mangabeira, Pedro A O; de Jesus, Raildo M; Pirovani, Carlos P; Ahnert, Dário; Baligar, Virupax C; Loguercio, Leandro L

2014-01-01

Seedlings of Theobroma cacao CCN 51 genotype were grown under greenhouse conditions and exposed to increasing concentrations of Cu (0.005, 1, 2, 4, 8, 16, and 32 mg Cu L(-1)) in nutrient solution. When doses were equal or higher than 8 mg Cu L(-1), after 24 h of treatment application, leaf gas exchange was highly affected and changes in chloroplasts thylakoids of leaf mesophyll cells and plasmolysis of cells from the root cortical region were observed. In addition, cell membranes of roots and leaves were damaged. In leaves, 96 h after treatments started, increases in the percentage of electrolyte leakage through membranes were observed with increases of Cu in the nutrient solution. Moreover, there was an increase in the concentration of thiobarbituric acid-reactive substances in roots due to lipid peroxidation of membranes. Chemical analysis showed that increases in Cu concentrations in vegetative organs of T. cacao increased with the increase of the metal in the nutrient solution, but there was a greater accumulation of Cu in roots than in shoots. The excess of Cu interfered in the levels of Mn, Zn, Fe, Mg, K, and Ca in different organs of T. cacao. Analysis of gene expression via RTq-PCR showed increased levels of MT2b, SODCyt, and PER-1 expression in roots and of MT2b, PSBA, PSBO, SODCyt, and SODChI in leaves. Hence, it was concluded that Cu in nutrient solution at doses equal or above 8 mg L(-1) significantly affected leaf gas exchange, cell ultrastructure, and transport of mineral nutrients in seedlings of this T. cacao genotype.

Response of the leaf photosynthetic rate to available nitrogen in erect panicle-type rice (Oryza sativa L. cultivar, Shennong265

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Chihiro Urairi

2016-07-01

Full Text Available Increasing the yield of rice per unit area is important because of the demand from the growing human population in Asia. A group of varieties called erect panicle-type rice (EP achieves very high yields under conditions of high nitrogen availability. Little is known, however, regarding the leaf photosynthetic capacity of EP, which may be one of the physiological causes of high yield. We analyzed the factors contributing to leaf photosynthetic rate (Pn and leaf mesophyll anatomy of Nipponbare, Takanari, and Shennong265 (a EP type rice cultivar varieties subjected to different nitrogen treatments. In the field experiment, Pn of Shennong265 was 33.8 μmol m−2 s−1 in the high-N treatment, and was higher than that of the other two cultivars because of its high leaf nitrogen content (LNC and a large number of mesophyll cells between the small vascular bundles per unit length. In Takanari, the relatively high value of Pn (31.5 μmol m−2 s−1 was caused by the high stomatal conductance (gs; .72 mol m−2 s−1 in the high-N treatment. In the pot experiment, the ratio of Pn/Ci to LNC, which may reflect mesophyll conductance (gm, was 20–30% higher in Nipponbare than in Takanari or Shennong265 in the high N availability treatment. The photosynthetic performance of Shennong265 might be improved by introducing the greater ratio of Pn/Ci to LNC found in Nipponbare and greater stomatal conductance found in Takanari.

Physiological responses of Vallisneria spiraslis L. induced by ...

African Journals Online (AJOL)

Jane

2011-07-25

Jul 25, 2011 ... and acts as a structural constituent in regulatory proteins, participates in .... of lipid peroxidation. The level of lipid peroxidation in plants was estimated by ..... Classification of nutrient emission sources in the. Vistula River ...

Monosaccharide analysis of succulent leaf tissue in Aloe

DEFF Research Database (Denmark)

Grace, Olwen Megan; Dzajic, Amra; Jäger, Anna

2013-01-01

in the genus Aloe using a predictive phylogenetic approach. Methodology – Monosaccharide composition was assessed in 31species, representing the morphological and taxonomic diversity of Aloe sensu stricto. Leaf mesophyll polysaccharides were partially hydrolysed in a trifluoroacetic acid (TFA)-SilA assay......Introduction – The succulent leaf mesophyll in Aloe species supports a burgeoning natural products industry, particularly in Africa. Comparative data necessary to prioritise species with economic potential have been lacking. Objective – To survey leaf mesophyll monosaccharide composition....... Oximes and trimethylsilyl ether products were detected by GC-MS. Constituent monosaccharides accounting for the greatest variation among species were identified by principal component analysis. Two plant DNA barcoding regions were sequenced in 28 of the sampled species and the resulting maximum...

Study of cell-differentiation and assembly of photosynthetic proteins during greening of etiolated Zea mays leaves using confocal fluorescence microspectroscopy at liquid-nitrogen temperature.

Science.gov (United States)

Shibata, Yutaka; Katoh, Wataru; Tahara, Yukari

2013-04-01

Fluorescence microspectroscopy observations were used to study the processes of cell differentiation and assemblies of photosynthesis proteins in Zea mays leaves under the greening process. The observations were done at 78K by setting the sample in a cryostat to avoid any undesired progress of the greening process during the measurements. The lateral and axial spatial resolutions of the system were 0.64μm and 4.4μm, respectively. The study revealed the spatial distributions of protochlorophyllide (PChld) in both the 632-nm-emitting and 655-nm-emitting forms within etiolated Zea mays leaves. The sizes of the fluorescence spots attributed to the former were larger than those of the latter, validating the assignment of the former and latter to the prothylakoid and prolamellar bodies, respectively. In vivo microspectroscopy observations of mature Zea mays leaves confirmed the different photosystem II (PS I)/photosystem I (PS II) ratio between the bundle sheath (BS) and mesophyll (MS) cells, which is specific for C4-plants. The BS cells in Zea mays leaves 1h after the initiation of the greening process tended to show fluorescence spectra at shorter wavelength side (at around 679nm) than the MS cells (at around 682nm). The 679-nm-emitting chlorophyll-a form observed mainly in the BS cells was attributed to putative precursor complexes to PS I. The BS cells under 3-h greening showed higher relative intensities of the PS I fluorescence band at around 735nm, suggesting the reduced PS II amount in the BS cells in this greening stage. Copyright © 2013 Elsevier B.V. All rights reserved.

Isolation of a strong Arabidopsis guard cell promoter and its potential as a research tool

Directory of Open Access Journals (Sweden)

Siegel Robert S

2008-02-01

gene silencing. It is also applicable to reduce specific gene expression in guard cells, providing a method for circumvention of limitations arising from genetic redundancy and lethality. These advances could be very useful for manipulating signaling pathways in guard cells and modifying plant performance under stress conditions. In addition, new guard cell and mesophyll cell-specific 23,000 gene microarray data are made publicly available here.

Effects of slag-based silicon fertilizer on rice growth and brown-spot resistance.

Science.gov (United States)

Ning, Dongfeng; Song, Alin; Fan, Fenliang; Li, Zhaojun; Liang, Yongchao

2014-01-01

It is well documented that slag-based silicon fertilizers have beneficial effects on the growth and disease resistance of rice. However, their effects vary greatly with sources of slag and are closely related to availability of silicon (Si) in these materials. To date, few researches have been done to compare the differences in plant performance and disease resistance between different slag-based silicon fertilizers applied at the same rate of plant-available Si. In the present study both steel and iron slags were chosen to investigate their effects on rice growth and disease resistance under greenhouse conditions. Both scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to examine the effects of slags on ultrastructural changes in leaves of rice naturally infected by Bipolaris oryaze, the causal agent of brown spot. The results showed that both slag-based Si fertilizers tested significantly increased rice growth and yield, but decreased brown spot incidence, with steel slag showing a stronger effect than iron slag. The results of SEM analysis showed that application of slags led to more pronounced cell silicification in rice leaves, more silica cells, and more pronounced and larger papilla as well. The results of TEM analysis showed that mesophyll cells of slag-untreated rice leaf were disorganized, with colonization of the fungus (Bipolaris oryzae), including chloroplast degradation and cell wall alterations. The application of slag maintained mesophyll cells relatively intact and increased the thickness of silicon layer. It can be concluded that applying slag-based fertilizer to Si-deficient paddy soil is necessary for improving both rice productivity and brown spot resistance. The immobile silicon deposited in host cell walls and papillae sites is the first physical barrier for fungal penetration, while the soluble Si in the cytoplasm enhances physiological or induced resistance to fungal colonization.

Cell-specific expression of tryptophan decarboxylase and 10-hydroxygeraniol oxidoreductase, key genes involved in camptothecin biosynthesis in Camptotheca acuminata Decne (Nyssaceae

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Santamaria Anna

2010-04-01

Full Text Available Abstract Background Camptotheca acuminata is a major natural source of the terpenoid indole alkaloid camptothecin (CPT. At present, little is known about the cellular distribution of the biosynthesis of CPT, which would be useful knowledge for developing new strategies and technologies for improving alkaloid production. Results The pattern of CPT accumulation was compared with the expression pattern of some genes involved in CPT biosynthesis in C. acuminata [i.e., Ca-TDC1 and Ca-TDC2 (encoding for tryptophan decarboxylase and Ca-HGO (encoding for 10-hydroxygeraniol oxidoreductase]. Both CPT accumulation and gene expression were investigated in plants at different degrees of development and in plantlets subjected to drought-stress. In all organs, CPT accumulation was detected in epidermal idioblasts, in some glandular trichomes, and in groups of idioblast cells localized in parenchyma tissues. Drought-stress caused an increase in CPT accumulation and in the number of glandular trichomes containing CPT, whereas no increase in epidermal or parenchymatous idioblasts was observed. In the leaf, Ca-TDC1 expression was detected in some epidermal cells and in groups of mesophyll cells but not in glandular trichomes; in the stem, it was observed in parenchyma cells of the vascular tissue; in the root, no expression was detected. Ca-TDC2 expression was observed exclusively in leaves of plantlets subjected to drought-stress, in the same sites described for Ca-TDC1. In the leaf, Ca-HGO was detected in all chlorenchyma cells; in the stem, it was observed in the same sites described for Ca-TDC1; in the root, no expression was detected. Conclusions The finding that the sites of CPT accumulation are not consistently the same as those in which the studied genes are expressed demonstrates an organ-to-organ and cell-to-cell translocation of CPT or its precursors.

Phakopsora euvitis Causes Unusual Damage to Leaves and Modifies Carbohydrate Metabolism in Grapevine

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Antonio F. Nogueira Júnior

2017-09-01

Full Text Available Asian grapevine rust (Phakopsora euvitis is a serious disease, which causes severe leaf necrosis and early plant defoliation. These symptoms are unusual for a strict biotrophic pathogen. This work was performed to quantify the effects of P. euvitis on photosynthesis, carbohydrates, and biomass accumulation of grapevine. The reduction in photosynthetic efficiency of the green leaf tissue surrounding the lesions was quantified using the virtual lesion concept (β parameter. Gas exchange and responses of CO2 assimilation to increasing intercellular CO2 concentration were analyzed. Histopathological analyses and quantification of starch were also performed on diseased leaves. Biomass and carbohydrate accumulation were quantified in different organs of diseased and healthy plants. Rust reduced the photosynthetic rate, and β was estimated at 5.78, indicating a large virtual lesion. Mesophyll conductance, maximum rubisco carboxylation rate, and regeneration of ribulose-1,5-bisphosphate dependent on electron transport rate were reduced, causing diffusive and biochemical limitations to photosynthesis. Hypertrophy, chloroplast degeneration of mesophyll cells, and starch accumulation in cells close to lesions were observed. Root carbohydrate concentration was reduced, even at low rust severity. Asian grapevine rust dramatically reduced photosynthesis and altered the dynamics of production and accumulation of carbohydrates, unlike strict biotrophic pathogens. The reduction in carbohydrate reserves in roots would support polyetic damage on grapevine, caused by a polycyclic disease.

Investigations on gamma ray induced chlorophyll variegated mutants

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Datta, S.K.; Dwivedi, A.K.; Banerji, B.K.

1995-01-01

Considering economic importance of chlorophyll variegation in floriculture trade an attempt was made for cytological, anatomical and biochemical analysis of four Bougainvillea and Lantana depressa chlorophyll variegated mutants for better and clear understanding of origin of chlorophyll variegation. No cytological evidence could be detected for their origin. Anatomical and biochemical examinations revealed that chlorophyll variegation in these mutants were due to changes in biosynthesis pathways and time of chlorophyll synthesis in palisade and spongy mesophyll cells. (author). 7 refs., 3 figs., 3 tabs

Localization of (photorespiration and CO2 re-assimilation in tomato leaves investigated with a reaction-diffusion model.

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Herman N C Berghuijs

Full Text Available The rate of photosynthesis depends on the CO2 partial pressure near Rubisco, Cc, which is commonly calculated by models using the overall mesophyll resistance. Such models do not explain the difference between the CO2 level in the intercellular air space and Cc mechanistically. This problem can be overcome by reaction-diffusion models for CO2 transport, production and fixation in leaves. However, most reaction-diffusion models are complex and unattractive for procedures that require a large number of runs, like parameter optimisation. This study provides a simpler reaction-diffusion model. It is parameterized by both leaf physiological and leaf anatomical data. The anatomical data consisted of the thickness of the cell wall, cytosol and stroma, and the area ratios of mesophyll exposed to the intercellular air space to leaf surfaces and exposed chloroplast to exposed mesophyll surfaces. The model was used directly to estimate photosynthetic parameters from a subset of the measured light and CO2 response curves; the remaining data were used for validation. The model predicted light and CO2 response curves reasonably well for 15 days old tomato (cv. Admiro leaves, if (photorespiratory CO2 release was assumed to take place in the inner cytosol or in the gaps between the chloroplasts. The model was also used to calculate the fraction of CO2 produced by (photorespiration that is re-assimilated in the stroma, and this fraction ranged from 56 to 76%. In future research, the model should be further validated to better understand how the re-assimilation of (photorespired CO2 is affected by environmental conditions and physiological parameters.

[Comparative leaf anatomy and phylogenetic relationships of 11 species of Laeliinae with emphasis on Brassavola (Orchidaceae)].

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Noguera-Savelli, Eliana; Jáuregui, Damelis

2011-09-01

Brassavola inhabits a wide altitude range and habitat types from Northern Mexico to Northern Argentina. Classification schemes in plants have normally used vegetative and floral characters, but when species are very similar, as in this genus, conflicts arise in species delimitation, and alternative methods should be applied. In this study we explored the taxonomic and phylogenetic value of the anatomical structure of leaves in Brassavola; as ingroup, seven species of Brassavola were considered, and as an outgroup Guarianthe skinneri, Laelia anceps, Rhyncholaelia digbyana and Rhyncholaelia glauca were evaluated. Leaf anatomical characters were studied in freehand cross sections of the middle portion with a light microscope. Ten vegetative anatomical characters were selected and coded for the phylogenetic analysis. Phylogenetic reconstruction was carried out under maximum parsimony using the program NONA through WinClada. Overall, Brassavola species reveal a wide variety of anatomical characters, many of them associated with xeromorphic plants: thick cuticle, hypodermis and cells of the mesophyll with spiral thickenings in the secondary wall. Moreover, mesophyll is either homogeneous or heterogeneous, often with extravascular bundles of fibers near the epidermis at both terete and flat leaves. All vascular bundles are collateral, arranged in more than one row in the mesophyll. The phylogenetic analysis did not resolve internal relationships of the genus; we obtained a polytomy, indicating that the anatomical characters by themselves have little phylogenetic value in Brassavola. We concluded that few anatomical characters are phylogenetically important; however, they would provide more support to elucidate the phylogenetic relantionships in the Orchidaceae and other plant groups if they are used in conjunction with morphological and/or molecular characters.

A simple and effective method to encapsulate tobacco mesophyll protoplasts to maintain cell viability

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Rong Lei

2015-01-01

• It is very convenient to change or collect the solution without mechanically disturbing the protoplasts. This simple and effective silica sol–gel/alginate two-step immobilization of protoplasts in Transwell has great potential for applications in genetic transformation, metabolite production, and migration assays.

A study on the isolation of protoplasts from mesophyll cells of Dendrobium Queen Pink

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Aqeel, R.; Zehra, M.; Kazmi, S. K.; Khan, S.

2016-01-01

Protoplasts were successfully isolated from one month old In vitro grown plantlets of Dendrobium cultivar Queen pink. The enzyme solution used was composed of 1 percent Cellulase Onozuka R-10, 0.5 percent Macerozyme R-10, 0.1 percent Pectinase, 0.3 M mannitol, 10 mM CaCl/sub 2/.2H/sub 2/O and 10 mM 2 (N-morpholino)-ethanesulfonic acid (MES) at pH 5.8. Protoplast highest yield with 15.7x104 protoplasts per 1.5 gm freshly chopped leaves were obtained when digested in enzyme solution for 4 hrs on a rotary shaker with an agitation speed of 45 rpm in dark conditions. Protoplasts were filtered with 45 micro m nylon sieve and washed with 0.3 M mannitol solution supplemented with 10 mM CaCl/sub 2/.2H/sub 2/O and 10 mM MES, and purified with 0.3 M sucrose solution gradient. Purification of protoplasts on a sucrose mannitol gradient yielded clean protoplasts that were free from debris. (author)

ISOLATION OF MESOPHYLL PROTOPLASTS FROM MEDITERRANEAN WOODY PLANTS FOR THE STUDY OF DNA INTEGRITY UNDER ABIOTIC STRESS

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Elena Kuzminsky

2016-08-01

Full Text Available Abiotic stresses have considerable negative impact on Mediterranean plant ecosystems and better comprehension of the genetic control of response and adaptation of trees to global changes is urgently needed. The Single Cell Gel Electrophoresis assay could be considered a good estimator of DNA damage in an individual eukaryotic cell. This method has been mainly employed in animal tissues, because the plant cell wall represents an obstacle for the extraction of nuclei; moreover, in Mediterranean woody species, especially in the sclerophyll plants, this procedure can be quite difficult because of the presence of sclerenchyma and hardened cells. On the other hand, these plants represent an interesting material to be studied because of the ability of these plants to tolerate abiotic stress. For instance, holm oak (Quercus ilex L. has been selected as the model plant to identify critical levels of O3 for Southern European forests. Consequently, a quantitative method for the evaluation of cell injury of leaf tissues of this species is required. Optimal conditions for high-yield nuclei isolation were obtained by using protoplast technology and a detailed description of the method is provided and discussed. White poplar (Populus alba L. was used as an internal control for protoplast isolation. Such a method has not been previously reported in newly fully developed leaves of holm oak. This method combined with Single Cell Gel Electrophoresis assay represents a new tool for testing the DNA integrity of leaf tissues in higher plants under stress conditions.

Photoprotection by foliar anthocyanins mitigates effects of boron toxicity in sweet basil (Ocimum basilicum).

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Landi, Marco; Guidi, Lucia; Pardossi, Alberto; Tattini, Massimiliano; Gould, Kevin S

2014-11-01

Boron (B) toxicity is an important agricultural problem in arid environments. Excess edaphic B compromises photosynthetic efficiency, limits growth and reduces crop yield. However, some purple-leafed cultivars of sweet basil (Ocimum basilicum) exhibit greater tolerance to high B concentrations than do green-leafed cultivars. We hypothesised that foliar anthocyanins protect basil leaf mesophyll from photo-oxidative stress when chloroplast function is compromised by B toxicity. Purple-leafed 'Red Rubin' and green-leafed 'Tigullio' cultivars, grown with high or negligible edaphic B, were given a photoinhibitory light treatment. Possible effects of photoabatement by anthocyanins were simulated by superimposing a purple polycarbonate filter on the green leaves. An ameliorative effect of light filtering on photosynthetic quantum yield and on photo-oxidative load was observed in B-stressed plants. In addition, when green protoplasts from both cultivars were treated with B and illuminated through a screen of anthocyanic protoplasts or a polycarbonate film which approximated cyanidin-3-O-glucoside optical properties, the degree of photoinhibition, hydrogen peroxide production, and malondialdehyde content were reduced. The data provide evidence that anthocyanins exert a photoprotective role in purple-leafed basil mesophyll cells, thereby contributing to improved tolerance to high B concentrations.

COMPARATIVE ANATOMICAL ASPECTS OF EUPHORBIA MILLI VAR. SPLENDENS (BOJER EX HOOK. URSCH & LEANDRI AND EUPHORBIA PULCHERRIMA WILLD. EX KLOTZSCH SPECIES LEAVES

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Rodica BERCU

2016-12-01

Full Text Available The paper presents a comparative study concerning the leaf structure of two Euphorbia species belonging to Euphorbiaceae family: Euphorbia milli var. splendens (Bojer ex Hook. Ursch & Leandri and Euphorbia pulcherrima Willd. ex Klotzsch. Anatomically, the leaves of the studied species are quite similar in the basic structure. The petiole has a single-layered epidermis, a collenchyma tissue – hypodermis - and three collateral bundles embedded in a basic tissue. Differences occur concerning the relationship between the collateral bundles. The Euphorbia pulcherrima bract petiole has almost the same structure as those of the leaf petiole. The blade is amphistomatic for Euphorbia milli var. splendes and hipostomatic for E. pulcherrima. The heterogeneous mesophyll is isobilateral in Euphorbia milli var. splendens blade and bifacial in E. pulcherrima. The vascular system of the mid rib is represented by one collateral bundle for both species, more developed in Euphorbia milli var. splendens blade. Differences appear concerning the epidermal cells cuticle thickness, the type of mesophyll, the abundance of the non-articulated laticifers and the development of the vascular system. The Euphorbia pulcherrima bract has the same organization plan structure as the leaf blade but some features differences occur.

Na+-Dependent High-Affinity Nitrate, Phosphate and Amino Acids Transport in Leaf Cells of the Seagrass Posidonia oceanica (L. Delile

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Lourdes Rubio

2018-05-01

Full Text Available Posidonia oceanica (L. Delile is a seagrass, the only group of vascular plants to colonize the marine environment. Seawater is an extreme yet stable environment characterized by high salinity, alkaline pH and low availability of essential nutrients, such as nitrate and phosphate. Classical depletion experiments, membrane potential and cytosolic sodium measurements were used to characterize the high-affinity NO3−, Pi and amino acids uptake mechanisms in this species. Net uptake rates of both NO3− and Pi were reduced by more than 70% in the absence of Na+. Micromolar concentrations of NO3− depolarized mesophyll leaf cells plasma membrane. Depolarizations showed saturation kinetics (Km = 8.7 ± 1 μM NO3−, which were not observed in the absence of Na+. NO3− induced depolarizations at increasing Na+ also showed saturation kinetics (Km = 7.2 ± 2 mM Na+. Cytosolic Na+ measured in P. oceanica leaf cells (17 ± 2 mM Na+ increased by 0.4 ± 0.2 mM Na+ upon the addition of 100 μM NO3−. Na+-dependence was also observed for high-affinity l-ala and l-cys uptake and high-affinity Pi transport. All together, these results strongly suggest that NO3−, amino acids and Pi uptake in P. oceanica leaf cells are mediated by high-affinity Na+-dependent transport systems. This mechanism seems to be a key step in the process of adaptation of seagrasses to the marine environment.

MORPHOLOGICAL ANALYSIS OF THE LEAVES OF Anacardium occidentale L.

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Glenda Quaresma Ramos

2016-03-01

Full Text Available In morphological studies are analyzed various parameters, ranging from macro scale through the micro scale to the nanometer scale, which contribute to the study of taxonomy, pharmacognosy, ecology, among others. Among the structures found in plants, the leaves are most organs analyzed. This study aimed to analyze morphological features of the leaves of the cashew tree, which is a plant of great commercial importance in Brazil. In this work we observed sinuous epidermal cells in the adaxial and abaxial, characterize their stomata in paracytic surrounded subsidiaries cells. On the abaxial surface the presence of glandular trichomes was observed; and cross-sectional analysis showed a single-layered epidermis with compact mesophyll and several layers of parenchyma cells. Keywords: leaf anatomy; cashew tree; optical microscopy.

Cd induced redistribution of elements within leaves of the Cd/Zn hyperaccumulator Thlaspi praecox as revealed by micro-PIXE

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Pongrac, Paula; Vogel-Mikus, Katarina; Vavpetic, Primoz; Tratnik, Janja; Regvar, Marjana; Simcic, Jurij; Grlj, Natasa; Pelicon, Primoz

2010-01-01

A detailed localisation of elements within leaf tissues of hydroponically grown Cd/Zn hyperaccumulator Thlaspi praecox (Brassicaceae) was determined by micro-PIXE at Jozef Stefan Institute (Ljubljana, Slovenia) in order to study accumulation patterns of Cd and other elements in the case of a single metal (Cd) pollution. Plants were treated with increasing concentrations of Cd in the solution (0 (control), 1, 10 and 100 μM). As expected, concentration of Cd in the leaves gradually increased with Cd concentration in the solution. In order to reveal the main Cd storage compartment space within the leaves a relative element distribution (pool) was calculated based on concentrations of elements in specific leaf tissues and their weight portions. Where present at detectable levels, Cd accumulated in the epidermal tissues (at 10 μM), but the contribution of epidermal pool decreased with increasing Cd concentration in solution (at 100 μM). The opposite was observed for the mesophyll pool. In addition, in Cd treated plants, a significant decrease in mesophyll Fe pool and an increase in the epidermal Fe pool were observed. Similar effect was seen for Mn pool at 100 μM Cd treatment accompanied by increasing Zn epidermal pool with increasing Cd in nutrient solution. Altogether these results indicate repartitioning of essential mesophyll cation pools (e.g., Fe, Mn and possibly Zn) when increasing Cd contents, that are instead more intensively stored in the epidermal cells. These results confirmed micro-PIXE as effective and powerful technique providing essential information on metal localisation, repartitioning and major elemental stores in plants on the tissue levels that were not accessible using classical analytical techniques and thus complementing our current understanding of plant metal tolerance mechanisms as a whole.

Growth of the C4 dicot Flaveria bidentis: photosynthetic acclimation to low light through shifts in leaf anatomy and biochemistry.

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Pengelly, Jasper J L; Sirault, Xavier R R; Tazoe, Youshi; Evans, John R; Furbank, Robert T; von Caemmerer, Susanne

2010-09-01

In C(4) plants, acclimation to growth at low irradiance by means of anatomical and biochemical changes to leaf tissue is considered to be limited by the need for a close interaction and coordination between bundle sheath and mesophyll cells. Here differences in relative growth rate (RGR), gas exchange, carbon isotope discrimination, photosynthetic enzyme activity, and leaf anatomy in the C(4) dicot Flaveria bidentis grown at a low (LI; 150 micromol quanta m(2) s(-1)) and medium (MI; 500 micromol quanta m(2) s(-1)) irradiance and with a 12 h photoperiod over 36 d were examined. RGRs measured using a 3D non-destructive imaging technique were consistently higher in MI plants. Rates of CO(2) assimilation per leaf area measured at 1500 micromol quanta m(2) s(-1) were higher for MI than LI plants but did not differ on a mass basis. LI plants had lower Rubisco and phosphoenolpyruvate carboxylase activities and chlorophyll content on a leaf area basis. Bundle sheath leakiness of CO(2) (phi) calculated from real-time carbon isotope discrimination was similar for MI and LI plants at high irradiance. phi increased at lower irradiances, but more so in MI plants, reflecting acclimation to low growth irradiance. Leaf thickness and vein density were greater in MI plants, and mesophyll surface area exposed to intercellular airspace (S(m)) and bundle sheath surface area per unit leaf area (S(b)) measured from leaf cross-sections were also both significantly greater in MI compared with LI leaves. Both mesophyll and bundle sheath conductance to CO(2) diffusion were greater in MI compared with LI plants. Despite being a C(4) species, F. bidentis is very plastic with respect to growth irradiance.

Studies on silica deposition in sugarcane (Saccharum spp. ) using scanning electron microscopy, energy-dispersive X-ray analysis, neutron activation analysis, and light microscopy

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Kaufman, P B; Takeoka, Y; Carlson, T J; Bigelow, W C; Jones, J D; Moore, P H; Ghosheh, N S [Michigan Univ., Ann Arbor (USA)

1979-06-01

Marked differences in silicon content in internodes of two sugarcane cultivars as revealed by neutron activation analysis, were closely correlated with number of silica cells per unit area in the epidermal system of the internodes of the two cultivars, as indicated by scanning electron microscopy and X-ray analysis. Light microscopy of epidermal peels showed that silica cells are capable of transmitting significantly more light through themselves than do other types of adjacent epidermal cells. This could be of great significance to total amount of carbon fixed by photosynthesizing mesophyll cells in leaves and cortical cells in internodes below the epidermis, especially in sugarcane cultivars with high densities of silica cells in their shoot epidermal systems. This has led to propose a window hypothesis, which indicates that silica cells in sugarcane, and in other grasses, act like windows in the epidermal system, allowing more light to be transmitted to photosynthetic tissue below than would occur if silica cells were absent.

Distribution and Translocation of 141Ce (III) in Horseradish

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Guo, Xiaoshan; Zhou, Qing; Lu, Tianhong; Fang, Min; Huang, Xiaohua

2007-01-01

Background and Aims Rare earth elements (REEs) are used in agriculture and a large amount of them contaminate the environment and enter foods. The distribution and translocation of 141Ce (III) in horseradish was investigated in order to help understand the biochemical behaviour and toxic mechanism of REEs in plants. Method The distribution and translocation of 141Ce (III) in horseradish were investigated using autoradiography, liquid scintillation counting (LSC) and electron microscopic autoradiography (EMARG) techniques. The contents of 141Ce (III) and nutrient elements were analysed using an inductively coupled plasma-atomic emission spectrometer (ICP-AES). Results The results from autoradiography and LSC indicated that 141Ce (III) could be absorbed by horseradish and transferred from the leaf to the leaf-stalk and then to the root. The content of 141Ce (III) in different parts of horseradish was as follows: root > leaf-stalk > leaf. The uptake rates of 141Ce (III) in horseradish changed with the different organs and time. The content of 141Ce (III) in developing leaves was greater than that in mature leaves. The results from EMARG indicated that 141Ce (III) could penetrate through the cell membrane and enter the mesophyll cells, being present in both extra- and intra-cellular deposits. The contents of macronutrients in horseradish were decreased by 141Ce (III) treatment. Conclusions 141Ce (III) can be absorbed and transferred between organs of horseradish with time, and the distribution was found to be different at different growth stages. 141Ce (III) can enter the mesophyll cells via apoplast and symplast channels or via plasmodesmata. 141Ce (III) can disturb the metabolism of macronutrients in horseradish. PMID:17921527

The cyclic nucleotide gated cation channel AtCNGC10 traffics from the ER via Golgi vesicles to the plasma membrane of Arabidopsis root and leaf cells

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Andres Marilou A

2007-09-01

Full Text Available Abstract Background The cyclic nucleotide-gated ion channels (CNGCs maintain cation homeostasis essential for a wide range of physiological processes in plant cells. However, the precise subcellular locations and trafficking of these membrane proteins are poorly understood. This is further complicated by a general deficiency of information about targeting pathways of membrane proteins in plants. To investigate CNGC trafficking and localization, we have measured Atcngc5 and Atcngc10 expression in roots and leaves, analyzed AtCNGC10-GFP fusions transiently expressed in protoplasts, and conducted immunofluorescence labeling of protoplasts and immunoelectron microscopic analysis of high pressure frozen leaves and roots. Results AtCNGC10 mRNA and protein levels were 2.5-fold higher in roots than leaves, while AtCNGC5 mRNA and protein levels were nearly equal in these tissues. The AtCNGC10-EGFP fusion was targeted to the plasma membrane in leaf protoplasts, and lightly labeled several intracellular structures. Immunofluorescence microscopy with affinity purified CNGC-specific antisera indicated that AtCNGC5 and AtCNGC10 are present in the plasma membrane of protoplasts. Immunoelectron microscopy demonstrated that AtCNGC10 was associated with the plasma membrane of mesophyll, palisade parenchyma and epidermal cells of leaves, and the meristem, columella and cap cells of roots. AtCNCG10 was also observed in the endoplasmic reticulum and Golgi cisternae and vesicles of 50–150 nm in size. Patch clamp assays of an AtCNGC10-GFP fusion expressed in HEK293 cells measured significant cation currents. Conclusion AtCNGC5 and AtCNGC10 are plasma membrane proteins. We postulate that AtCNGC10 traffics from the endoplasmic reticulum via the Golgi apparatus and associated vesicles to the plasma membrane. The presence of the cation channel, AtCNGC10, in root cap meristem cells, cell plate, and gravity-sensing columella cells, combined with the previously reported

Poinsettia protoplasts - a simple, robust and efficient system for transient gene expression studies

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Pitzschke Andrea

2012-05-01

Full Text Available Abstract Background Transient gene expression systems are indispensable tools in molecular biology. Yet, their routine application is limited to few plant species often requiring substantial equipment and facilities. High chloroplast and chlorophyll content may further impede downstream applications of transformed cells from green plant tissue. Results Here, we describe a fast and simple technique for the high-yield isolation and efficient transformation (>70% of mesophyll-derived protoplasts from red leaves of the perennial plant Poinsettia (Euphorbia pulccherrima. In this method no particular growth facilities or expensive equipments are needed. Poinsettia protoplasts display an astonishing robustness and can be employed in a variety of commonly-used downstream applications, such as subcellular localisation (multi-colour fluorescence or promoter activity studies. Due to low abundance of chloroplasts or chromoplasts, problems encountered in other mesophyll-derived protoplast systems (particularly autofluorescence are alleviated. Furthermore, the transgene expression is detectable within 90 minutes of transformation and lasts for several days. Conclusions The simplicity of the isolation and transformation procedure renders Poinsettia protoplasts an attractive system for transient gene expression experiments, including multi-colour fluorescence, subcellular localisation and promoter activity studies. In addition, they offer hitherto unknown possibilities for anthocyan research and industrial applications.

Clustering and cellular distribution characteristics of virus particles of Tomato spotted wilt virus and Tomato zonate spot virus in different plant hosts.

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Zhang, Zhongkai; Zheng, Kuanyu; Dong, Jiahong; Fang, Qi; Hong, Jian; Wang, Xifeng

2016-01-19

Tomato spotted wilt virus (TSWV) and Tomato zonate spot virus (TZSV) are the two dominant species of thrip-transmitted tospoviruses, cause significant losses in crop yield in Yunnan and its neighboring provinces in China. TSWV and TZSV belong to different serogroup of tospoviruses but induce similar symptoms in the same host plant species, which makes diagnostic difficult. We used different electron microscopy preparing methods to investigate clustering and cellular distribution of TSWV and TZSV in the host plant species. Negative staining of samples infected with TSWV and TZSV revealed that particles usually clustered in the vesicles, including single particle (SP), double particles clustering (DPC), triple particles clustering (TPC). In the immunogold labeling negative staining against proteins of TZSV, the antibodies against Gn protein were stained more strongly than the N protein. Ultrathin section and high pressure freeze (HPF)-electron microscopy preparations revealed that TSWV particles were distributed in the cisternae of endoplasmic reticulum (ER), filamentous inclusions (FI) and Golgi bodies in the mesophyll cells. The TSWV particles clustered as multiple particles clustering (MPC) and distributed in globular viroplasm or cisternae of ER in the top leaf cell. TZSV particles were distributed more abundantly in the swollen membrane of ER in the mesophyll cell than those in the phloem parenchyma cells and were not observed in the top leaf cell. However, TZSV virions were mainly present as single particle in the cytoplasm, with few clustering as MPC. In this study, we identified TSWV and TZSV particles had the distinct cellular distribution patterns in the cytoplasm from different tissues and host plants. This is the first report of specific clustering characteristics of tospoviruses particles as well as the cellular distribution of TSWV particles in the FI and globular viroplasm where as TZSV particles inside the membrane of ER. These results indicated that

Somatic embryogenesis and in vitro plant regeneration from pejibaye adult plant leaf primordia Embriogênese somática e regeneração de plantas in vitro a partir de primórdios foliares de pupunheiras adultas

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Marcílio de Almeida

2006-09-01

Full Text Available The aim of this work was to evaluate a protocol for plant regeneration by means of somatic embryos obtained from isolated adult pejibaye leaf primordia, and to describe histological origin of embryos and morphogenetic response. Explants were cultivated in modified MS medium. Mesophyll parenchymatous cells originated meristemoids (preembryonic complex formation induced with 7.1 µM BAP in the first two subculture periods. After polarized structures with 12.9 µM NAA and 3.55 µM BAP were formed in the third subculture, somatic embryos developed and regenerated normal plants. The mesophyll parenchymatous cells display high capacity of direct response to the auxin and cytokinin.O objetivo deste trabalho foi avaliar um protocolo de regeneração de plantas por meio de embriões somáticos, obtidos a partir de primórdios foliares de pupunheiras adultas e identificar a origem histológica dos embriões e descrever as etapas morfogenéticas. Os explantes foram cultivados em meio MS modificado. Células parenquimáticas do mesofilo originaram meristemóides com BAP (7,1 µM nos dois primeiros períodos de subcultura. A polarização das estruturas ocorreu com ANA (12,9 µM e BAP (3,55 µM no terceiro período de subcultura. Meristemóides se desenvolveram em embriões somáticos, regenerando plantas normais. As células parenquimáticas do mesofilo apresentam elevada capacidade de resposta direta à auxina e à citocinina.

Tissue localization of u.v.-B-screening pigments and of chalcone synthase mRNA in needles of Scots pine seedlings

International Nuclear Information System (INIS)

Schnitzler, J.P.; Jungblut, T.P.; Heller, W.; Köfferlein, M.; Hutzler, P.; Heinzmann, U.; Schmelzer, E.; Ernst, D.; Langebartels, C.; Sandermann, H. Jr.

1996-01-01

Epidermal tissue was isolated from Scots pine (Pinus sylvestris L.) needles by enzymatic digestion in order to study tissue distribution of u.v.-B-screening pigments. Up to 90% of the needle content of a group of diacylated flavonol glycosides that were structurally closely related was found in the epidermal layer. Among these metabolites, 3'',6''-di-para-coumaroyl-isoquercitrin and 3'',6''-di-para-coumaroyl-astragalin were the main u.v.-B-induced compounds in cotyledons and primary needles, respectively. However, catechin and astragalin (kaempferol 3-glucoside), two non-acylated flavonoid metabolites, were only observed in total needle extracts, and at levels independent of u.v.-B treatment. According to this metabolite distribution, the mRNA of chalcone synthase, the key enzyme to flavonoids, was found in epidermal and mesophyll as well as vascular tissues. The major alkaliextractable wall-bound phenolic metabolites, astragalin, 4-coumaric acid, and ferulic acid, a minor component of the cell wall, were also found exclusively in the epidermal layer. These compounds were not stimulated by u.v.-B irradiation within the experimental period. Staining of needle cross sections and epidermal layer preparations with Naturstoffreagenz A confirmed the specific localization of wall-bound astragalin in the outer wall of the epidermal layer. Model calculations of u.v.-B absorptions at 300 nm of soluble and cell-wall-bound metabolites of the epidermal layer revealed an almost complete shielding of the mesophyll tissue from u.v.-B radiation

Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation.

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Okumura, Masaki; Inoue, Shin-Ichiro; Kuwata, Keiko; Kinoshita, Toshinori

2016-05-01

Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. © 2016 American Society of Plant Biologists. All Rights Reserved.

Secretory cavities and volatiles of Myrrhinium atropurpureum Schott var. atropurpureum (Myrtaceae): an endemic species collected in the restingas of Rio de Janeiro, Brazil.

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Victório, Cristiane Pimentel; Moreira, Claudio B; Souza, Marcelo da Costa; Sato, Alice; Arruda, Rosani do Carmo de Oliveira

2011-07-01

In this study, we investigated the leaf anatomy and the composition of volatiles in Myrrhinium atropurpureum var. atropurpureum endemic to Rio de Janeiro restingas. Particularly, leaf secretory structures were described using light microscopy, and histochemical tests were performed from fresh leaves to localize the secondary metabolites. To observe secretory cavities, fixed leaf samples were free-hand sectioned. To evaluate lipophilic compounds and terpenoids the following reagents were employed: Sudans III and IV, Red oil O and Nile blue. Leaf volatiles were characterized by gas chromatography after hydrodistillation (HD) or simultaneous distillation-extraction (SDE). Leaf analysis showed several cavities in mesophyll that are the main sites of lipophilic and terpenoid production. Monoterpenes, which represented more than 80% of the major volatiles, were characterized mainly by alpha- and beta-pinene and 1,8-cineole. In order to provide tools for M. atropurpureum identification, the following distinguishing characteristics were revealed by the following data: 1) adaxial face clear and densely punctuated by the presence of round or ellipsoidal secretory cavities randomly distributed in the mesophyll; 2) the presence of cells overlying the upper neck cells of secretory cavities; 3) the presence of numerous paracytic stomata distributed on the abaxial leaf surface, but absent in vein regions and leaf margin; and 4) non-glandular trichomes on both leaf surfaces. Our study of the compounds produced by the secretory cavities of M. atropurpureum led us to conclude that volatile terpenoid class are the main secretory compounds and that they consist of a high concentration of monoterpenes, which may indicate the phytotherapeutic importance of this plant.

Pharmacognostic study of Chamaecyparis lawsoniana (Murr. Parl.: A drug used in Homoeopathy

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Anshu Rathi

2015-01-01

Full Text Available The pharmacognostic profile of crude drug has a key role in standardization for quality, purity and drug identification. The present study deals with pharmacognostic evaluation of aerial part of Chamaecyparis lawsoniana (Murr. Parl. a drug used in homoeopathic system of medicine for diverse clinical uses such as terrible pain in stomach, tumors, keloid, warts and lipoma of thigh. The study includes collection, identification, macroscopy, microscopy and organoleptic characteristics of aerial part of Chamaecyparis lawsoniana. Anatomically the leaf is distinguishable into a layer of the epidermis followed by parenchymatous mesophyll and resin duct in the parenchymatous cortex. Powder microscopy shows the presence of epidermal cells, parenchymatous cells and tracheids. These observations may be used as pharmacopoeial standards for identification of Cha maecyparis lawsoniana.

Investigation of the site-specific accumulation of catechins in the tea plant (Camellia sinensis (L.) O. Kuntze) via vanillin-HCl staining.

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Liu, Yajun; Gao, Liping; Xia, Tao; Zhao, Lei

2009-11-11

Histochemical staining using vanillin-HCl is a potential tool to identify the site-specific accumulation of catechins in the tea plant (Camellia sinensis (L.) O. Kuntze). Using this technique revealed that catechins existed ubiquitously in all inspected tissues in young tea leaf, but the distribution was concentrated in the vascular bundle and palisade tissue, whereas the large parenchyma cells of the main vein contained lower amounts of catechins. At the subcellular level, catechins were located mainly in the chloroplasts of mesophyll cells and in the vessel wall. In young stems, catechins could be detected in most cells except the parenchyma cells of the pith and the cortex, whereas, in roots, catechins could be detected only in those cells surrounding the pericycle. Moreover, differing distributions of catechins were found in calluses cultivated in darkness and light. On the basis of HPLC analyses, six main types of catechins were present in tea leaves, stems, calluses, and chloroplasts; however, roots contained only epicatechin.

Immunolocalization of IAA and ABA in roots and needles of radiata pine (Pinus radiata) during drought and rewatering.

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De Diego, N; Rodríguez, J L; Dodd, I C; Pérez-Alfocea, F; Moncaleán, P; Lacuesta, M

2013-05-01

Anatomical, physiological and phytohormonal changes involved in drought tolerance were examined in different Pinus radiata D. Don breeds subjected to soil drying and rewatering. Breeds with the smallest stomatal chamber size had the lowest transpiration rate and the highest intrinsic water-use efficiency. Xylem cell size was positively correlated with leaf hydraulic conductance and needle indole-3-acetic acid (IAA) concentrations, whereas transpiration rate was negatively correlated with needle abscisic acid (ABA) levels. Since these two phytohormones seem important in regulating the P. radiata drought response, they were simultaneously immunolocalized in roots and needles of the most tolerant breed (P. radiata var. radiata × var. cedrosensis) during two sequential drought cycles and after rewatering. During drought, IAA was unequally distributed into the pointed area of the needle cross-section and mainly located in mesophyll and vascular tissue cells of needles, possibly inducing needle epinasty, whereas ABA was principally located in guard cells, presumably to elicit stomata closure. In the roots, at the end of the first drought cycle, while strong IAA accumulation was observed in the cortex, ABA levels decreased probably due to translocation to the leaves. Rewatering modified the distribution of both IAA and ABA in the needles, causing an accumulation principally in vascular tissue, with residual concentrations in mesophyll, likely favouring the acclimatization of the plants for further drought cycles. Contrarily, in the roots IAA and ABA were located in the exodermis, a natural barrier that regulates the phytohormone translocation to other plant tissues and hormone losses to the soil solution after rewatering. These results confirm that immunolocalization is an efficient tool to understand the translocation of IAA and ABA in plants subjected to different water stress situations, and clarify their role in regulating physiological responses such as stomata

Bundle-sheath leakiness in C4 photosynthesis: a careful balancing act between CO2 concentration and assimilation.

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Kromdijk, Johannes; Ubierna, Nerea; Cousins, Asaph B; Griffiths, Howard

2014-07-01

Crop species with the C4 photosynthetic pathway are generally characterized by high productivity, especially in environmental conditions favouring photorespiration. In comparison with the ancestral C3 pathway, the biochemical and anatomical modifications of the C4 pathway allow spatial separation of primary carbon acquisition in mesophyll cells and subsequent assimilation in bundle-sheath cells. The CO2-concentrating C4 cycle has to operate in close coordination with CO2 reduction via the Calvin-Benson-Bassham (CBB) cycle in order to keep the C4 pathway energetically efficient. The gradient in CO2 concentration between bundle-sheath and mesophyll cells facilitates diffusive leakage of CO2. This rate of bundle-sheath CO2 leakage relative to the rate of phosphoenolpyruvate carboxylation (termed leakiness) has been used to probe the balance between C4 carbon acquisition and subsequent reduction as a result of environmental perturbations. When doing so, the correct choice of equations to derive leakiness from stable carbon isotope discrimination (Δ(13)C) during gas exchange is critical to avoid biased results. Leakiness responses to photon flux density, either short-term (during measurements) or long-term (during growth and development), can have important implications for C4 performance in understorey light conditions. However, recent reports show leakiness to be subject to considerable acclimation. Additionally, the recent discovery of two decarboxylating C4 cycles operating in parallel in Zea mays suggests that flexibility in the transported C4 acid and associated decarboxylase could also aid in maintaining C4/CBB balance in a changing environment. In this paper, we review improvements in methodology to estimate leakiness, synthesize reports on bundle-sheath leakiness, discuss different interpretations, and highlight areas where future research is necessary. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology

Functional characterization of secondary wall deposition regulating transcription factors MusaVND2 and MusaVND3 in transgenic banana plants.

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Negi, Sanjana; Tak, Himanshu; Ganapathi, T R

2016-03-01

NAM, ATAF, and CUC (NAC) domain-containing proteins are plant-specific transcription factors involved in stress responses and developmental regulation. MusaVND2 and MusaVND3 are vascular-related NAC domain-containing genes encoding for nuclear-localized proteins. The transcript level of MusaVND2 and MusaVND3 are gradually induced after induction of lignification conditions in banana embryogenic cells. Banana embryogenic cells differentiated to tracheary element-like cells after overexpression of MusaVND2 and MusaVND3 with a differentiation frequency of 63.5 and 23.4 %, respectively, after ninth day. Transgenic banana plants overexpressing either of MusaVND2 or MusaVND3 showed ectopic secondary wall deposition as well as transdifferentiation of cells into tracheary elements. Transdifferentiation to tracheary element-like cells was observed in cortical cells of corm and in epidermal and mesophyll cells of leaves of transgenic plants. Elevated levels of lignin and crystalline cellulose were detected in the transgenic banana lines than control plants. The results obtained are useful for understanding the molecular regulation of secondary wall development in banana.

Analysis of Nanobody-Epitope Interactions in Living Cells via Quantitative Protein Transport Assays.

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Früholz, Simone; Pimpl, Peter

2017-01-01

Over the past few decades, quantitative protein transport analyses have been used to elucidate the sorting and transport of proteins in the endomembrane system of plants. Here, we have applied our knowledge about transport routes and the corresponding sorting signals to establish an in vivo system for testing specific interactions between soluble proteins.Here, we describe the use of quantitative protein transport assays in tobacco mesophyll protoplasts to test for interactions occurring between a GFP-binding nanobody and its GFP epitope. For this, we use a secreted GFP-tagged α-amylase as a reporter together with a vacuolar-targeted RFP-tagged nanobody. The interaction between these proteins is then revealed by a transport alteration of the secretory reporter due to the interaction-triggered attachment of the vacuolar sorting signal.

C4 photosynthetic machinery: insights from maize chloroplast proteomics

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Qi eZhao

2013-04-01

Full Text Available C4 plants exhibit much higher CO2 assimilation rates than C3 plants. The specialized differentiation of mesophyll cell (M and bundle sheath cell (BS type chloroplasts is unique to C4 plants and improves photosynthesis efficiency. Maize (Zea mays is an important crop and model with C4 photosynthetic machinery. Current high-throughput quantitative proteomics approaches (e.g., 2DE, iTRAQ, and shotgun proteomics have been employed to investigate maize chloroplast structure and function. These proteomic studies have provided valuable information on C4 chloroplast protein components, photosynthesis, and other metabolic mechanisms underlying chloroplast biogenesis, stromal and membrane differentiation, as well as response to salinity, high/low temperature, and light stress. This review presents an overview of proteomics advances in maize chloroplast biology.

CO2 sensing and CO2 regulation of stomatal conductance: advances and open questions

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Engineer, Cawas; Hashimoto-Sugimoto, Mimi; Negi, Juntaro; Israelsson-Nordstrom, Maria; Azoulay-Shemer, Tamar; Rappel, Wouter-Jan; Iba, Koh; Schroeder, Julian

2015-01-01

Guard cells form epidermal stomatal gas exchange valves in plants and regulate the aperture of stomatal pores in response to changes in the carbon dioxide (CO2) concentration in leaves. Moreover, the development of stomata is repressed by elevated CO2 in diverse plant species. Evidence suggests that plants can sense CO2 concentration changes via guard cells and via mesophyll tissues in mediating stomatal movements. We review new discoveries and open questions on mechanisms mediating CO2-regulated stomatal movements and CO2 modulation of stomatal development, which together function in CO2-regulation of stomatal conductance and gas exchange in plants. Research in this area is timely in light of the necessity of selecting and developing crop cultivars which perform better in a shifting climate. PMID:26482956

Alleviatory effects of silicon on the foliar micromorphology and anatomy of rice (Oryza sativa L.) seedlings under simulated acid rain.

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Ju, Shuming; Wang, Liping; Zhang, Cuiying; Yin, Tingchao; Shao, Siliang

2017-01-01

Silicon (Si) is a macroelement in plants. The biological effects and mitigation mechanisms of silicon under environmental stress have become hot topics. The main objectives of this study were to elucidate the roles of Si in alleviating the effects on the phenotype, micromorphology and anatomy of the leaves of rice seedlings under acid rain stress. The results indicated that the combined or single effects of Si and simulated acid rain (SAR) stress on rice roots depended on the concentration of Si and the intensity of the SAR stress. The combined or single effects of the moderate concentration of Si (2.0 mM) and light SAR (pH 4.0) enhanced the growth of the rice leaves and the development of the mesophyll cells, and the combined effects were stronger than those of the single treatments. The high concentration of Si (4.0 mM) and severe SAR (pH 3.0 or 2.0) exerted deleterious effects. The incorporation of Si (2.0 or 4.0 mM) into SAR at pH values of 3.0 or 2.0 promoted rice leaf growth, decreased necrosis spots, maintained the structure and function of the mesophyll cells, increased the epicuticular wax content and wart-like protuberance (WP) density, and improved the stomatal characteristics of the leaves of rice seedlings more than the SAR only treatments. The alleviatory effects observed with a moderate concentration of Si (2.0 mM) were better than the effects obtained with the high concentration of Si (4.0 mM). The alleviatory effects were due to the enhancement of the mechanical barriers in the leaf epidermis.

Alleviatory effects of silicon on the foliar micromorphology and anatomy of rice (Oryza sativa L.) seedlings under simulated acid rain

Science.gov (United States)

Ju, Shuming; Wang, Liping; Zhang, Cuiying; Yin, Tingchao; Shao, Siliang

2017-01-01

Silicon (Si) is a macroelement in plants. The biological effects and mitigation mechanisms of silicon under environmental stress have become hot topics. The main objectives of this study were to elucidate the roles of Si in alleviating the effects on the phenotype, micromorphology and anatomy of the leaves of rice seedlings under acid rain stress. The results indicated that the combined or single effects of Si and simulated acid rain (SAR) stress on rice roots depended on the concentration of Si and the intensity of the SAR stress. The combined or single effects of the moderate concentration of Si (2.0 mM) and light SAR (pH 4.0) enhanced the growth of the rice leaves and the development of the mesophyll cells, and the combined effects were stronger than those of the single treatments. The high concentration of Si (4.0 mM) and severe SAR (pH 3.0 or 2.0) exerted deleterious effects. The incorporation of Si (2.0 or 4.0 mM) into SAR at pH values of 3.0 or 2.0 promoted rice leaf growth, decreased necrosis spots, maintained the structure and function of the mesophyll cells, increased the epicuticular wax content and wart-like protuberance (WP) density, and improved the stomatal characteristics of the leaves of rice seedlings more than the SAR only treatments. The alleviatory effects observed with a moderate concentration of Si (2.0 mM) were better than the effects obtained with the high concentration of Si (4.0 mM). The alleviatory effects were due to the enhancement of the mechanical barriers in the leaf epidermis. PMID:29065171

Reduced Tonoplast Fast-Activating and Slow-Activating Channel Activity Is Essential for Conferring Salinity Tolerance in a Facultative Halophyte, Quinoa1[C][W][OA

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Bonales-Alatorre, Edgar; Shabala, Sergey; Chen, Zhong-Hua; Pottosin, Igor

2013-01-01

Halophyte species implement a “salt-including” strategy, sequestering significant amounts of Na+ to cell vacuoles. This requires a reduction of passive Na+ leak from the vacuole. In this work, we used quinoa (Chenopodium quinoa) to investigate the ability of halophytes to regulate Na+-permeable slow-activating (SV) and fast-activating (FV) tonoplast channels, linking it with Na+ accumulation in mesophyll cells and salt bladders as well as leaf photosynthetic efficiency under salt stress. Our data indicate that young leaves rely on Na+ exclusion to salt bladders, whereas old ones, possessing far fewer salt bladders, depend almost exclusively on Na+ sequestration to mesophyll vacuoles. Moreover, although old leaves accumulate more Na+, this does not compromise their leaf photochemistry. FV and SV channels are slightly more permeable for K+ than for Na+, and vacuoles in young leaves express less FV current and with a density unchanged in plants subjected to high (400 mm NaCl) salinity. In old leaves, with an intrinsically lower density of the FV current, FV channel density decreases about 2-fold in plants grown under high salinity. In contrast, intrinsic activity of SV channels in vacuoles from young leaves is unchanged under salt stress. In vacuoles of old leaves, however, it is 2- and 7-fold lower in older compared with young leaves in control- and salt-grown plants, respectively. We conclude that the negative control of SV and FV tonoplast channel activity in old leaves reduces Na+ leak, thus enabling efficient sequestration of Na+ to their vacuoles. This enables optimal photosynthetic performance, conferring salinity tolerance in quinoa species. PMID:23624857

Search for C4 developmental mutants in Panicum maximum Jacq

International Nuclear Information System (INIS)

Fladung, M.

2001-01-01

small leaves and reached a total plant height of maximal 0.6 meter. In leaf sections, it was characterized by an almost lack of the small veins surrounded by four bundle sheath cells. The leaf lamina of the variant abs showed several alterations, including doublets of veins, veins without bundle sheath, additional bundle sheath cells outside the veins or large bundle sheath cells participating in two bundle sheaths. Also the distribution of phloem and xylem cells within the bundles were quite altered in the variant compared to the wildtype. The leaves were greener, with a higher than normal chlorophyll content and with longitudinal veins not perfectly straight but following a wavy path on the leaf lamina. Compared with wild type plants the phenotype of the mbl mutant was less erect and had pending leaves because of the absence of the main midrib. In wild type leaves the midrib was represented by an enlargement of the mesophyll parenchyma which included parenchymatous and sclerenchymatous cells. This structure was absent in mutant leaves, only small irregular files of parenchymatous cells were present at the base of the leaf lamina. The florets of this mutant had no carpel but one or two additional stamen. The variant var1 had a variegated phenotype with stripes of yellow-green and white tissues alternating the leaf laminae. In yellow-green sectors the chloroplasts were absent only in bundle sheath cells, which supports the hypothesis of different ways of development of bundle sheath and mesophyll cell chloroplasts. The adjacent mesophyll cells were less pigmented than similar ones present in non-variant sectors. In white sectors, the chloroplasts were absent both in bundle sheath and mesophyll cells. The variant was partially fertile. Seed germination was 30 to 40%, and despite the unknown portion of apomictic seeds, 65 produced white, 20 green and 34 variegated seedlings out of a sample of 119 germinated seeds. Analysis of segregation of these green and variegated

Somatic hybridization in Citrus: navel orange (C. sinensis Osb.) and grapefruit (C. paradisi Macf.).

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Ohgawara, T; Kobayashi, S; Ishii, S; Yoshinaga, K; Oiyama, I

1989-11-01

Protoplasts of navel orange, isolated from embryogenic nucellar cell suspension culture, were fused with protoplasts of grapefruit isolated from leaf tissue. The fusion products were cultured in the hormone-free medium containing 0.6 M sucrose. Under the culture conditions, somatic embryogenesis of navel orange protoplasts was suppressed, while cell division of grapefruit mesophyll protoplasts was not induced. Six embryoids were obtained and three lines regenerated to complete plants through embryogenesis. Two of the regenerated lines exhibited intermediate morphological characteristics of the parents in the leaf shape. Chromosome counts showed that these regenerated plants had expected 36 chromosomes (2n=2x=18 for each parent). The rDNA analysis using biotin-labeled rRNA probes confirmed the presence of genomes from both parents in these plants. This somatic hybridization system would be useful for the practical Citrus breeding.

Transport and concentration of abscisic acid (ABA) and auxin (IAA) in deciduous and coniferous trees. Transport und Gehalt von Abscisinsaeure (ABA) und Auxin (IAA) in Laub- und Nadelblaettern

Energy Technology Data Exchange (ETDEWEB)

Hartung, W.

1988-09-01

Abscisic acid and indoleacetic acid were chosen to examine whether intact deciduous and coniferous tissues from spruce, hemlock fir, spinage, barley and sorrel or isolated mesophyll protoplasts from barley and closing cell preparations from Valerianella locusta are affected by sulphur dioxide in terms of changes in the concentration, transportation and distribution of such plant hormones. The distribution of phytohormones like ABA and IAA over the individual cell compartments is determined by the different pH gradients of the latter. Owing to their acidity these hormones are accumulated in alkaline cell inclusion bodies like chloroplasts and cytosol. Potentially acid air pollutants like SO{sub 2} and NO{sub x} lead to acidification of previously alkaline cell compartments, due to which fact the cellular pH gradients are reduced. This, in turn, gives rise to a redistribution of phytohormones to the effect that certain target cells such as closing cells of leaves or meristem cells come under the influence of altered hormone concentrations and compositions. This is bound to affect the processes controlling the development, growth and stress behaviour of plants. (orig.) With 55 refs., 2 tabs., 16 figs.

Different patterns of vein loading of exogenous [14C]sucrose in leaves of pisum sativum and coleus blumei

International Nuclear Information System (INIS)

Turgeon, R.; Wimmers, L.E.

1988-01-01

Vein loading of exogenous [ 14 C]sucrose was studied using short uptake and wash periods to distinguish between direct loading into veins and loading via mesophyll tissue. Mature leaf tissue of Pisum sativum L. cv Little Marvel, or Coleus blumei Benth. cv Candidum, was abraded and leaf discs were floated on [ 14 C]sucrose solution for 1 or 2 minutes. Discs were then washed for 1 to 30 min either at room temperature or in the cold and were frozen, lyophilized, and autoradiographed. In P. sativum, veins were clearly labeled after 1 minute uptake and 1 minute wash periods. Autoradiographic images did not change appreciably with longer times of uptake or wash. Vein loading was inhibited by p-chloromercuribenzenesulfonic acid. These results indicate that uptake of exogenous sucrose occurs directly into the veins in this species. When C. blumei leaf discs were floated on [ 14 C]sucrose for 2 minutes and washed in the cold, the mesophyll was labeled but little, if any, minor vein loading occurred. When discs were labeled for 2 minutes and washed at room temperature, label was transferred from the mesophyll to the veins within minutes. These results indicate that there may be different patterns of phloem loading of photosynthetically derived sucrose in these two species

Burkholderia phytofirmans PsJN reduces damages to freezing temperature in Arabidopsis thaliana

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Fan eSU

2015-10-01

Full Text Available Several plant growth-promoting rhizobacteria (PGPR are known to improve plant tolerance to multiple stresses, including low temperatures. However, mechanisms underlying this protection are still poorly understood. The aim of this study was to evaluate the role of the endophytic PGPR, Burkholderia phytofirmans strain PsJN (Bp PsJN, on Arabidopsis thaliana cold tolerance using photosynthesis parameters as physiological markers.Under standard conditions, our results indicated that Bp PsJN inoculation led to growth promotion of Arabidopsis plants without significant modification on photosynthesis parameters and chloroplast organization. However, bacterial colonization induced a cell wall strengthening in the mesophyllImpact of inoculation modes (either on seeds or by soil irrigation and their effects overnight at 0, -1 or -3°C, were investigated by following photosystem II (PSII activity and gas exchanges. Following low temperatures stress, a decrease of photosynthesis parameters was observed. In addition, during three consecutive nights or days at -1°C, PSII activity was monitored. Pigment contents, RuBisCO protein abundance, expression of several genes including RbcS, RbcL, CBF1, CBF2, CBF3, ICE1, COR15a, and COR78 were evaluated at the end of exposure. To assess the impact of the bacteria on cell ultrastructure under low temperatures, microscopic observations were achieved. Results indicated that freezing treatment induced significant changes in PSII activity as early as the first cold day, whereas the same impact on PSII activity was observed only during the third cold night. The significant effects conferred by PsJN were differential accumulation of pigments, and reduced expression of RbcL and COR78. Microscopical observations showed an alteration/disorganization in A. thaliana leaf mesophyll cells independently of the freezing treatments. The presence of bacteria during the three successive nights or days did not significantly improved A

Variable content and distribution of arabinogalactan proteins in banana (Musa spp.) under low temperature stress.

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Yan, Yonglian; Takáč, Tomáš; Li, Xiaoquan; Chen, Houbin; Wang, Yingying; Xu, Enfeng; Xie, Ling; Su, Zhaohua; Šamaj, Jozef; Xu, Chunxiang

2015-01-01

Information on the spatial distribution of arabinogalactan proteins (AGPs) in plant organs and tissues during plant reactions to low temperature (LT) is limited. In this study, the extracellular distribution of AGPs in banana leaves and roots, and their changes under LT stress were investigated in two genotypes differing in chilling tolerance, by immuno-techniques using 17 monoclonal antibodies against different AGP epitopes. Changes in total classical AGPs in banana leaves were also tested. The results showed that AGP epitopes recognized by JIM4, JIM14, JIM16, and CCRC-M32 antibodies were primarily distributed in leaf veins, while those recognized by JIM8, JIM13, JIM15, and PN16.4B4 antibodies exhibited predominant sclerenchymal localization. Epitopes recognized by LM2, LM14, and MAC207 antibodies were distributed in both epidermal and mesophyll cells. Both genotypes accumulated classical AGPs in leaves under LT treatment, and the chilling tolerant genotype contained higher classical AGPs at each temperature treatment. The abundance of JIM4 and JIM16 epitopes in the chilling-sensitive genotype decreased slightly after LT treatment, and this trend was opposite for the tolerant one. LT induced accumulation of LM2- and LM14-immunoreactive AGPs in the tolerant genotype compared to the sensitive one, especially in phloem and mesophyll cells. These epitopes thus might play important roles in banana LT tolerance. Different AGP components also showed differential distribution patterns in banana roots. In general, banana roots started to accumulate AGPs under LT treatment earlier than leaves. The levels of AGPs recognized by MAC207 and JIM13 antibodies in the control roots of the tolerant genotype were higher than in the chilling sensitive one. Furthermore, the chilling tolerant genotype showed high immuno-reactivity against JIM13 antibody. These results indicate that several AGPs are likely involved in banana tolerance to chilling injury.

Genetic and Developmental Basis for Increased Leaf Thickness in the Arabidopsis Cvi Ecotype

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Viktoriya Coneva

2018-03-01

Full Text Available Leaf thickness is a quantitative trait that is associated with the ability of plants to occupy dry, high irradiance environments. Despite its importance, leaf thickness has been difficult to measure reproducibly, which has impeded progress in understanding its genetic basis, and the associated anatomical mechanisms that pattern it. Here, we used a custom-built dual confocal profilometer device to measure leaf thickness in the Arabidopsis Ler × Cvi recombinant inbred line population and found statistical support for four quantitative trait loci (QTL associated with this trait. We used publically available data for a suite of traits relating to flowering time and growth responses to light quality and show that three of the four leaf thickness QTL coincide with QTL for at least one of these traits. Using time course photography, we quantified the relative growth rate and the pace of rosette leaf initiation in the Ler and Cvi ecotypes. We found that Cvi rosettes grow slower than Ler, both in terms of the rate of leaf initiation and the overall rate of biomass accumulation. Collectively, these data suggest that leaf thickness is tightly linked with physiological status and may present a tradeoff between the ability to withstand stress and rapid vegetative growth. To understand the anatomical basis of leaf thickness, we compared cross-sections of Cvi and Ler leaves and show that Cvi palisade mesophyll cells elongate anisotropically contributing to leaf thickness. Flow cytometry of whole leaves show that endopolyploidy accompanies thicker leaves in Cvi. Overall, our data suggest that mechanistically, an altered schedule of cellular events affecting endopolyploidy and increasing palisade mesophyll cell length contribute to increase of leaf thickness in Cvi. Ultimately, knowledge of the genetic basis and developmental trajectory leaf thickness will inform the mechanisms by which natural selection acts to produce variation in this adaptive trait.

Leaf Structure and Taxonomy of Petunia and Calibrachoa (Solanaceae

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Claudia dos Reis

2002-03-01

Full Text Available We studied the leaf anatomy of sixteen species of Calibrachoa and eight species of Petunia. In Calibrachoa leaves, the vascular bundles sheath (endodermis was formed by parenchymatous developed cells, different from those of the mesophyll. In Petunia, this sheath did not show a marked morphological differentiation. The Calibrachoa leaves could be separated according to the type of leaf margins, the distribution of the stomata on leaf surfaces, the organization of the mesophyll and the morphology of the trichomes. Based on these results, an indented dichotomous identification key was elaborated for the species of the genus Calibrachoa.Foram estudados, sob o ponto de vista anatômico, os limbos foliares de dezesseis espécies de Calibrachoa Llav. & Lex. e de oito espécies de Petunia Juss. (Solanaceae. Em Calibrachoa, a bainha que envolve os feixes vasculares (endoderme é formada por células desenvolvidas e distintas das do mesofilo. Em Petunia, esta bainha não apresenta diferenciação morfológica marcante. As folhas das espécies de Calibrachoa foram separadas entre si levando-se em conta a distribuição dos estômatos nas faces foliares, a organização do mesofilo, o tipo de bordo e a morfologia dos tricomas. Com base nesses resultados, foi elaborada uma chave dicotômica indentada de identificação para as espécies do gênero Calibrachoa.

Secretory structures of Ipomoea asarifolia: anatomy and histochemistry

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Fabiano M. Martins

2012-02-01

Full Text Available Ipomoea asarifolia (Desr. Roem. & Schult., Convolvulaceae, is a weed that infests agricultural areas and is toxic to cattle. In spite of its toxicity, the leaves of this plant are used in traditional remedies in the state of Bahia, Brazil. The present work describes the leaf anatomy of I. asarifolia and characterizes the exudates of its secretory structures. The leaves have a unistratified epidermis composed of ordinary cells with straight to slightly sinuous anticlinal walls and thin cuticles. Paracytic stomata are found on both surfaces of the leaves at the same level as the ordinary epidermal cells. Trichomes producing polysaccharide secretions occur on the petiole and leaf blade and are considered colleters. The mesophyll is dorsiventral and the vascular bundle of the central vein is bicollateral. Two opposed nectaries occur on the petiole near the leaf blade. Each nectary is composed of a small canal with internal ramifications and numerous secretory trichomes. The laticiferous glands are articulated, not anastomosed, and are composed of large diameter cells with thin cell walls. The secretions of the laticiferous glands are lipidic.

Arundina graminifolia var. revoluta (Arethuseae, Orchidaceae) has fern-type rheophyte characteristics in the leaves.

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Yorifuji, Eri; Ishikawa, Naoko; Okada, Hiroshi; Tsukaya, Hirokazu

2015-03-01

Morphological and molecular variation between Arundina graminifolia var. graminifolia and the dwarf variety, A. graminifolia var. revoluta, was examined to assess the validity of their taxonomic characteristics and genetic background for identification. Morphological analysis in combination with field observations indicated that A. graminifolia var. revoluta is a rheophyte form of A. graminifolia characterized by narrow leaves, whereas the other morphological characteristics described for A. graminifolia var. revoluta, such as smaller flowers and short stems, were not always accompanied by the narrower leaf phenotype. Molecular analysis based on matK sequences indicated that only partial differentiation has occurred between A. graminifolia var. graminifolia and A. graminifolia var. revoluta. Therefore, we should consider the rheophyte form an ecotype rather than a variety. Anatomical observations of the leaves revealed that the rheophyte form of A. graminifolia possessed characteristics of the rheophytes of both ferns and angiosperms, such as narrower palisade tissue cells and thinner spongy tissue cells, as well as fewer cells in the leaf-width direction and fewer mesophyll cell layers.

A major trade-off between structural and photosynthetic investments operative across plant and needle ages in three Mediterranean pines.

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Kuusk, Vivian; Niinemets, Ülo; Valladares, Fernando

2018-04-01

Pine (Pinus) species exhibit extensive variation in needle shape and size between juvenile (primary) and adult (secondary) needles (heteroblasty), but few studies have quantified the changes in needle morphological, anatomical and chemical traits upon juvenile-to-adult transition. Mediterranean pines keep juvenile needles longer than most other pines, implying that juvenile needles play a particularly significant role in seedling and sapling establishment in this environment. We studied needle anatomical, morphological and chemical characteristics in juvenile and different-aged adult needles in Mediterranean pines Pinus halepensis Mill., Pinus pinea L. and Pinus nigra J. F. Arnold subsp. salzmannii (Dunal) Franco hypothesizing that needle anatomical modifications upon juvenile-to-adult transition lead to a trade-off between investments in support and photosynthetic tissues, and that analogous changes occur with needle aging albeit to a lower degree. Compared with adult needles, juvenile needles of all species were narrower with 1.6- to 2.4-fold lower leaf dry mass per unit area, and had ~1.4-fold thinner cell walls, but needle nitrogen content per dry mass was similar among plant ages. Juvenile needles also had ~1.5-fold greater mesophyll volume fraction, ~3-fold greater chloroplast volume fraction and ~1.7-fold greater chloroplast exposed to mesophyll exposed surface area ratio, suggesting overall greater photosynthetic activity. Changes in needle traits were similar in aging adult needles, but the magnitude was generally less than the changes upon juvenile to adult transition. In adult needles, the fraction in support tissues scaled positively with known ranking of species tolerance of drought (P. halepensis > P. pinea > P. nigra). Across all species, and needle and plant ages, a negative correlation between volume fractions of mesophyll and structural tissues was observed, manifesting a trade-off between biomass investments in different needle functions. These

Effects of simulated acid rain on leaf anatomy and micromorphology of Genipa americana L. (Rubiaceae

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Bruno Francisco Sant'Anna-Santos

2006-03-01

Full Text Available Experiments were conducted in order to characterize the injuries on leaf structure and micromorphology of G. americana and evaluate the degree of susceptibility of this species to simulated acid rain. Plants were exposed to acid rain (pH 3.0 for ten consecutive days. Control plants were submitted only to distilled water (pH 6.0. Leaf tissue was sampled and fixed for light and scanning electron microscopy. Necrotic interveinal spots on the leaf blade occurred. Epidermis and mesophyll cells collapse, hypertrophy of spongy parenchyma cells, accumulation of phenolic compounds and starch grains were observed in leaves exposed to acid rain. The micromorphological analysis showed, in necrotic areas, plasmolized guard cells and cuticle rupture. Epidermal and mesophyll cells alterations occurred before symptoms were visualized in the leaves. These results showed the importance of anatomical data for precocious diagnosis injury and to determine the sensitivity of G. americana to acid rain.Experimentos foram conduzidos para avaliar o grau de susceptibilidade e determinar as injúrias causadas pela chuva ácida simulada na anatomia e micromorfologia foliar de Genipa americana. Plantas foram expostas à chuva com pH 3,0 durante 10 dias consecutivos. No tratamento controle utilizou-se apenas água destilada (pH 6,0. Amostras foliares foram coletadas e fixadas para microscopia de luz e eletrônica de varredura. Foram observados nas folhas expostas à chuva ácida: necroses pontuais intervenais, colapso das células do mesofilo e da epiderme; hipertrofia do parênquima lacunoso e acúmulo de compostos fenólicos e grãos de amido. A análise micromorfológica evidenciou, nas áreas necrosadas, plasmólise das células-guarda e ruptura da cutícula e da crista estomática. Alterações anatômicas ocorreram antes que sintomas visuais fossem observados nas folhas. Estes resultados comprovam a importância de dados anatômicos na diagnose precoce da injúria e na

Amborella trichopoda, plasmodesmata, and the evolution of phloem loading.

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Turgeon, Robert; Medville, Richard

2011-01-01

Phloem loading is the process by which photoassimilates synthesized in the mesophyll cells of leaves enter the sieve elements and companion cells of minor veins in preparation for long distance transport to sink organs. Three loading strategies have been described: active loading from the apoplast, passive loading via the symplast, and passive symplastic transfer followed by polymer trapping of raffinose and stachyose. We studied phloem loading in Amborella trichopoda, a premontane shrub that may be sister to all other flowering plants. The minor veins of A. trichopoda contain intermediary cells, indicative of the polymer trap mechanism, forming an arc on the abaxial side and subtending a cluster of ordinary companion cells in the interior of the veins. Intermediary cells are linked to bundle sheath cells by highly abundant plasmodesmata whereas ordinary companion cells have few plasmodesmata, characteristic of phloem that loads from the apoplast. Intermediary cells, ordinary companion cells, and sieve elements form symplastically connected complexes. Leaves provided with (14)CO(2) translocate radiolabeled sucrose, raffinose, and stachyose. Therefore, structural and physiological evidence suggests that both apoplastic and polymer trapping mechanisms of phloem loading operate in A. trichopoda. The evolution of phloem loading strategies is complex and may be difficult to resolve.

Studies on anatomical characters indicating C3 and C4 photosynthetic metabolism in the genus Boerhavia L. (Nyctaginaceae

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Abdulwakeel Ayokun-nun Ajao

2017-08-01

Full Text Available The C3 and C4 photosynthetic pathways in dicotyledons were investigated with the four species of Boerhavia occurring in Nigeria using light microscopy. The study is not yet well reported on dicotyledons as done for monocotyledons. The features cross-examined were stomata index, stomata size, inter-stomatal distance, stomatal density, interveinal distance, intercellular air spaces, leaf thickness, mesophyll thickness, Kranz tissue, one cell distant count criterion, maximum lateral cell count criterion, vein density and vein distance. Based on these features, these species (B. erecta, B. coccinea and B. repens were grouped into C4 while B. diffusa was grouped as a C3 plant. In particular, interveinal distance less than 166µm and maximum lateral count ranging 2 to 6 will help in grouping C4 dicotyledons species while those that were greater than these values are useful in grouping C3 and plants.

Somatic hybridization of sexually incompatible petunias: Petunia parodii, Petunia parviflora.

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Power, J B; Berry, S F; Chapman, J V; Cocking, E C

1980-01-01

Somatic hybrid plants were regenerated following the fusion of leaf mesophyll protoplasts of P. parodii with those isolated from a nuclear-albino mutant of P. parviflora. Attempts at sexual hybridization of these two species repeatedly failed thus confirming their previously established cross-incompatibility. Selection of somatic hybrid plants was possible since protoplasts of P. parodii would not develop beyond the cell colony stage, whilst those of the somatic hybrid and albino P. parviflora produced calluses. Green somatic hybrid calluses were visible against a background of albino cells/calluses, and upon transfer to regeneration media gave rise to shoots. Shoots and the resultant flowering plants were confirmed as somatic hybrids based on their growth habit, floral pigmentation and morphology, leaf hair structure, chromosome number and Fraction 1 protein profiles. The relevance of such hybrid material for the development of new, and extensively modified cultivars, is discussed.

Anatomical structure and surface micromorphology of tomatillo leaf and flower (Physalis ixocarpa Brot., Solanaceae

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Barbara Dyki

2014-01-01

Full Text Available Tomatillo (Physalis ixocarpa Brot. is a newly introduced cultivated plant in Poland. Its anatomy was investigated in light and scanning electron microscopes. Tomatillo adult leaf had one layer of palisade parenchyma. The 1-2 cell layers of spongy parenchyma situated just below the palisade parenchyma showed large, tightly packed cells with great druses. The remaining spongy parenchyma was built of cells showing several extensions. Peculiarity of the sepals were the stomata situated on columns or hills formed of many cells. The petals had a very loose mesophyl. Their adaxial epidermis was composed of papillate cells. Such structure of the petal epidermis probably contributes to light dispersion and prevents glittering. There were several types of trichomes on the leaves, sepals and petals, some of them glandular and some simple. The large, very ramified, dendritic trichomes situated on the petals at the entry to the ovary might eventually protect it against excessive drying. The pollen grain was spherical, three-colpate. The style had a hollow channel inside. The stigma was of a wet, pa-pillate type. Sometimes thorny trichomes were found among papillae.

Relationship between Hexokinase and the Aquaporin PIP1 in the Regulation of Photosynthesis and Plant Growth

Science.gov (United States)

Kelly, Gilor; Sade, Nir; Attia, Ziv; Secchi, Francesca; Zwieniecki, Maciej; Holbrook, N. Michele; Levi, Asher; Alchanatis, Victor; Moshelion, Menachem; Granot, David

2014-01-01

Increased expression of the aquaporin NtAQP1, which is known to function as a plasmalemma channel for CO2 and water, increases the rate of both photosynthesis and transpiration. In contrast, increased expression of Arabidopsis hexokinase1 (AtHXK1), a dual-function enzyme that mediates sugar sensing, decreases the expression of photosynthetic genes and the rate of transpiration and inhibits growth. Here, we show that AtHXK1 also decreases root and stem hydraulic conductivity and leaf mesophyll CO2 conductance (g m). Due to their opposite effects on plant development and physiology, we examined the relationship between NtAQP1 and AtHXK1 at the whole-plant level using transgenic tomato plants expressing both genes simultaneously. NtAQP1 significantly improved growth and increased the transpiration rates of AtHXK1-expressing plants. Reciprocal grafting experiments indicated that this complementation occurs when both genes are expressed simultaneously in the shoot. Yet, NtAQP1 had only a marginal effect on the hydraulic conductivity of the double-transgenic plants, suggesting that the complementary effect of NtAQP1 is unrelated to shoot water transport. Rather, NtAQP1 significantly increased leaf mesophyll CO2 conductance and enhanced the rate of photosynthesis, suggesting that NtAQP1 facilitated the growth of the double-transgenic plants by enhancing mesophyll conductance of CO2. PMID:24498392

Relationship between hexokinase and the aquaporin PIP1 in the regulation of photosynthesis and plant growth.

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Gilor Kelly

Full Text Available Increased expression of the aquaporin NtAQP1, which is known to function as a plasmalemma channel for CO₂ and water, increases the rate of both photosynthesis and transpiration. In contrast, increased expression of Arabidopsis hexokinase1 (AtHXK1, a dual-function enzyme that mediates sugar sensing, decreases the expression of photosynthetic genes and the rate of transpiration and inhibits growth. Here, we show that AtHXK1 also decreases root and stem hydraulic conductivity and leaf mesophyll CO₂ conductance (g(m. Due to their opposite effects on plant development and physiology, we examined the relationship between NtAQP1 and AtHXK1 at the whole-plant level using transgenic tomato plants expressing both genes simultaneously. NtAQP1 significantly improved growth and increased the transpiration rates of AtHXK1-expressing plants. Reciprocal grafting experiments indicated that this complementation occurs when both genes are expressed simultaneously in the shoot. Yet, NtAQP1 had only a marginal effect on the hydraulic conductivity of the double-transgenic plants, suggesting that the complementary effect of NtAQP1 is unrelated to shoot water transport. Rather, NtAQP1 significantly increased leaf mesophyll CO₂ conductance and enhanced the rate of photosynthesis, suggesting that NtAQP1 facilitated the growth of the double-transgenic plants by enhancing mesophyll conductance of CO₂.

An evaluation of the effects of exogenous ethephon, an ethylene releasing compound, on photosynthesis of mustard (Brassica juncea cultivars that differ in photosynthetic capacity

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Khan NA

2004-12-01

Full Text Available Abstract Background The stimulatory effect of CO2 on ethylene evolution in plants is known, but the extent to which ethylene controls photosynthesis is not clear. Studies on the effects of ethylene on CO2 metabolism have shown conflicting results. Increase or inhibition of photosynthesis by ethylene has been reported. To understand the physiological processes responsible for ethylene-mediated changes in photosynthesis, stomatal and mesophyll effects on photosynthesis and ethylene biosynthesis in response to ethephon treatment in mustard (Brassica juncea cultivars differing in photosynthetic capacity were studied. Results The effects of ethephon on photosynthetic rate (PN, stomatal conductance (gS, carbonic anhydrase (CA activity, 1-aminocyclopropane carboxylic acid synthase (ACS activity and ethylene evolution were similar in both the cultivars. Increasing ethephon concentration up to 1.5 mM increased PN, gS and CA maximally, whereas 3.0 mM ethephon proved inhibitory. ACS activity and ethylene evolution increased with increasing concentrations of ethephon. The corresponding changes in gs and CA activity suggest that the changes in photosynthesis in response to ethephon were triggered by altered stomatal and mesophyll processes. Stomatal conductance changed in parallel with changes in mesophyll photosynthetic properties. In both the cultivars ACS activity and ethylene increased up to 3.0 mM ethephon, but 1.5 mM ethephon caused maximum effects on photosynthetic parameters. Conclusion These results suggest that ethephon affects foliar gas exchange responses. The changes in photosynthesis in response to ethephon were due to stomatal and mesophyll effects. The changes in gS were a response maintaining stable intercellular CO2 concentration (Ci under the given treatment in both the cultivars. Also, the high photosynthetic capacity cultivar, Varuna responded less to ethephon than the low photosynthetic capacity cultivar, RH30. The photosynthetic

From reconstruction to C>4 metabolic engineering: A case study for overproduction of polyhydroxybutyrate in bioenergy grasses

DEFF Research Database (Denmark)

Gomes de Oliveira Dal'Molin, Cristiana; Quek, Lake-Ee; Saa, Pedro A.

2018-01-01

bundle sheath (B) and mesophyll (M) across the day and night cycle. The C4 leaf model was used to explore how amenable polyhydroxybutyrate (PHB) production is with these four compartments working cooperatively. A strategic pattern of metabolite conversion and exchange emerged from a systems-level network......The compartmentalization of C4 plants increases photosynthetic efficiency, while constraining how material and energy must flow in leaf tissues. To capture this metabolic phenomenon, a generic plant metabolic reconstruction was replicated into four connected spatiotemporal compartments, namely...... that has very few constraints imposed; mainly the sequential two-step carbon capture in mesophyll, then bundle sheath and photosynthesis during the day only. The building of starch reserves during the day and their mobilization during the night connects day and night metabolism. Flux simulations revealed...

Subcellular Lipid Droplets in Vanilla Leaf Epidermis and Avocado Mesocarp Are Coated with Oleosins of Distinct Phylogenic Lineages1[OPEN

Science.gov (United States)

2016-01-01

Subcellular lipid droplets (LDs) in diverse plant cells and species are coated with stabilizing oleosins of at least five phylogenic lineages and perform different functions. We examined two types of inadequately studied LDs for coated oleosins and their characteristics. The epidermis but not mesophyll of leaves of vanilla (Vanilla planifolia) and most other Asparagales species contained solitary and clustered LDs (avocado (Persea americana) and other Lauraceae species possessed large LDs, which likely function in attracting animals for seed dispersal. They contained transcripts of oleosin of a novel M phylogenic lineage. Each avocado mesocarp fatty cell possessed one to several large LDs (5 to 20 μm) and at their periphery, numerous small LDs (<0.5 μm). Immuno-confocal laser scanning microscopy revealed that oleosin was present mostly on the small LDs. LDs in isolated fractions coalesced rapidly, and the fraction contained oleosin and several other proteins and triacylglycerols as the main lipids. These two new types of oleosin-LDs exemplify the evolutionary plasticity of oleosins-LDs in generating novel functions in diverse cell types and species. PMID:27208281

Change of mitotic behavior and ultra structure of 'Fuju' (Citrus reticulata Blanco) stem-apex clones after space flight

International Nuclear Information System (INIS)

Wu Rujian; Huang Jinghao; Wen Shouxing; Cai Zijian; Luo Tuyan; Chen Liangfeng; Wang Zhouwen

2011-01-01

By using conventional squash stain technique and ultrathin sectioning technique, the effects of space flight on mitotic behavior and ultrastructure were studied in the shoot apical meristem of 'Fuju' (Citrus reticulata Blanco), which had been carried by China's seed-breeding satellite, Shijian-8. The results showed that space flight had effect on the mutagenesis of stem-apical meristem. Abnormal mitosis with various degrees had been detected in 13 mutant clones, of which mitotic aberrations in clone '08004' were significantly higher than the others. The aberration rate of numerical abnormalities of chromosomes at metaphase, lagging chromosome, micronucleus, C-spindle, S-spindle and polyarch spindle in the clone '08004' was 0.34%, 0.669%, 0.86%, 0.17%, 1.20% and 1.03%, respectively. The ultrastructure of mesophyll cell in most clones was unchanged, but nucleus chromatin agglutination, chloroplast thylakoid disintegrated, autophagosome appeared, cell vacuolated, plasmolysis and the formation of apoptotic body were found in the clone '08004', suggesting that programmed cell death (PCD) Nevertheless, no change in the mitochondrial structure was observed until terminal phase of PCD. (authors)

Effects of shading on Vallisneria natans (Lour.) H. Hara growth

DEFF Research Database (Denmark)

Fox, Anthony David; Meng, F; Shen, X

2013-01-01

, but dry weight fruit production was significantly reduced at 90% shading. We conclude that above 50% surface shading, V. natans plants suffer reductions in accumulated biomass and investment in sexual reproduction. We contend that recent expansions in the extent of the native floating water chestnut Trapa...

The ERECTA gene regulates plant transpiration efficiency in Arabidopsis.

Science.gov (United States)

Masle, Josette; Gilmore, Scott R; Farquhar, Graham D

2005-08-11

Assimilation of carbon by plants incurs water costs. In the many parts of the world where water is in short supply, plant transpiration efficiency, the ratio of carbon fixation to water loss, is critical to plant survival, crop yield and vegetation dynamics. When challenged by variations in their environment, plants often seem to coordinate photosynthesis and transpiration, but significant genetic variation in transpiration efficiency has been identified both between and within species. This has allowed plant breeders to develop effective selection programmes for the improved transpiration efficiency of crops, after it was demonstrated that carbon isotopic discrimination, Delta, of plant matter was a reliable and sensitive marker negatively related to variation in transpiration efficiency. However, little is known of the genetic controls of transpiration efficiency. Here we report the isolation of a gene that regulates transpiration efficiency, ERECTA. We show that ERECTA, a putative leucine-rich repeat receptor-like kinase (LRR-RLK) known for its effects on inflorescence development, is a major contributor to a locus for Delta on Arabidopsis chromosome 2. Mechanisms include, but are not limited to, effects on stomatal density, epidermal cell expansion, mesophyll cell proliferation and cell-cell contact.

Estudo anátomo-morfológico de dicotiledôneas das dunas de Salvador - Bahia: Borreria cymosa Cham. et Schl. e Chiococca brachiata R. et P. (Rubiaceae Morpho-anatomic studies of dicotiledons from Salvador - Bahia dunes: Borreria cymosa Chamo et Schl. and Chiococca brachiata R. et P. (Rubiaceae

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Elzeni Diladelfo de Gusmão

1992-07-01

Full Text Available Prosseguindo as pesquisas sobre a vegetação das Dunas do Abaeté, Salvador, Bahia, apresentam-se as Rubiaceae: Borreria cymosa e Chiococca brachiata. Cortes histológicos à mão livre, com lâminas de barbear, seguindo-se as técnicas de rotina. Estudaram-se as epidermes, mesofilo, nervura central, bordo e pecíolo. Contaram-se os estômatos por área foliar. Folhas dorsiventrais e hipoestomáticas, glabras, cutícula adaxial espessa e brilhante. Numerosas gotas lipídicas, esclereídeos no córtex do pecíolo e nervura central. Borreria: tem epiderme abaxial com paredes fortemente sinuosas e estrias epicuticulares; células incolores, subjacentes à epiderme adaxial; mesofilo com drusas e ráfides de oxolato de cálcio, bainhas de células volumosas envolvendo todos os feixes. Chiococca: epiderme de paredes curvas, cutícula formando "flanges" ao nível da nervura mediana, bordo e pecíolo. Pequenas células cheias de cloroplastos envolvem feixes menores. Clorênquima denso. Conclui-se que as plantas apresentam caracteres de adaptação ao meio xérico. As características anatômicas tanto quanto ao morfológicas individualizam os taxons.Following studies of the vegetation from the dunes of Abaeté, Salvador, Bahia, the Rubiaceae Borreria cymosa and Chiococca brachiata are described. Free hand histological were made with razor blades, following routine procedures. The epidermis, mesophyll, central nerve, margin and petiole were studied. Stomata per area of leaf were counted. Dorsiventral and hypoestomatic leaves, glabrous, adaxial cuticle heavy anda brillant. Numerous lipid droplets, sclereids in cortex of petiole and central nerve. Borreria: has abaxial epidermis with strongly sinuos walls and striated; cells colorless, subjacent to adaxial epidermis; mesophyll with druses and raphides of calcium oxalate, sheaths of volumous cells wrapping all the bundles. Chiococca: epiderm with curved walls, cuticle forming "flanges" at the level of the

Viral protein synthesis in cowpea mosaic virus infected protoplasts

International Nuclear Information System (INIS)

Rottier, P.

1980-01-01

Some aspects of cowpea mosaic virus (CPMV) multiplication in cowpea mesophyll protoplasts were studied. The detection and characterization of proteins whose synthesis is induced or is stimulated upon virus infection was performed with the aid of radioactive labelling. (Auth.)

The utility of Bambusoideae (Poaceae, Poales leaf blade anatomy for identification and systematics

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T. D. Leandro

Full Text Available Abstract Bambusoideae is a diverse subfamily that includes herbaceous (Olyreae and woody (Arundinarieae and Bambuseae bamboos. Species within Bambusae are particularly difficult to identify due to their monocarpic lifecycle and the often long durations between mass flowering events; whereas the herbaceous bamboos are pluricarpic, but often are found with no reproductive structures. The leaf blade anatomy of 16 sympatric species of native Brazilian bamboos (Olyreae and Bambuseae from the Atlantic Rainforest was studied in order to detect useful features for their identification. All the studied species share the following features: epidermis with a single stratum of cells; adaxial bulliform cells; mesophyll with arm cells, rosette cells, and fusoid cells; and collateral vascular bundles. Herbaceous bamboos share two features: papillae scattered on the abaxial surface and parallel-sided arrays of bulliform cells; whereas woody bamboos share: centrally organized papillae and fan-shaped arrays of bulliform cells. Also within the woody bamboos, intercostal fibers and a midrib with only one vascular bundle (simple midrib characterize the subtribe Arthrostylidiinae; whereas a midrib with more than one vascular bundle (complex midrib and a stomatal apparatus with two pappilae per subsidiary cell characterize the subtribe Chusqueinae. There are also diagnostic features for the sampled species, such as: papillae shape, and the outline and structure of the midrib. An identification key for all the studied species is provided based on the anatomical features.

Dynamics of Chloroplast Translation during Chloroplast Differentiation in Maize.

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Prakitchai Chotewutmontri

2016-07-01

Full Text Available Chloroplast genomes in land plants contain approximately 100 genes, the majority of which reside in polycistronic transcription units derived from cyanobacterial operons. The expression of chloroplast genes is integrated into developmental programs underlying the differentiation of photosynthetic cells from non-photosynthetic progenitors. In C4 plants, the partitioning of photosynthesis between two cell types, bundle sheath and mesophyll, adds an additional layer of complexity. We used ribosome profiling and RNA-seq to generate a comprehensive description of chloroplast gene expression at four stages of chloroplast differentiation, as displayed along the maize seedling leaf blade. The rate of protein output of most genes increases early in development and declines once the photosynthetic apparatus is mature. The developmental dynamics of protein output fall into several patterns. Programmed changes in mRNA abundance make a strong contribution to the developmental shifts in protein output, but output is further adjusted by changes in translational efficiency. RNAs with prioritized translation early in development are largely involved in chloroplast gene expression, whereas those with prioritized translation in photosynthetic tissues are generally involved in photosynthesis. Differential gene expression in bundle sheath and mesophyll chloroplasts results primarily from differences in mRNA abundance, but differences in translational efficiency amplify mRNA-level effects in some instances. In most cases, rates of protein output approximate steady-state protein stoichiometries, implying a limited role for proteolysis in eliminating unassembled or damaged proteins under non-stress conditions. Tuned protein output results from gene-specific trade-offs between translational efficiency and mRNA abundance, both of which span a large dynamic range. Analysis of ribosome footprints at sites of RNA editing showed that the chloroplast translation machinery

Electron transport, pep carboxylase activity, and maximal net co2 assimilation exhibit coordinated and proportional decline with loss of hydraulic conductance during water stress in Zea mays

Science.gov (United States)

Efforts to improve the photosynthetic performance of species are presently focused on leaf-level traits (e.g., quantum efficiency, mesophyll osmoregulation, stress protein regulation). Here, we emphasize that efforts to improve plant performance in arid environments would benefit from also consider...

The regulation of starch accumulation in Panicum maximum Jacq ...

African Journals Online (AJOL)

... decrease the starch level. These observations are discussed in relation to the photosynthetic characteristics of P. maximum. Keywords: accumulation; botany; carbon assimilation; co2 fixation; growth conditions; mesophyll; metabolites; nitrogen; nitrogen levels; nitrogen supply; panicum maximum; plant physiology; starch; ...

Use of sediment CO2 by submersed rooted plants

DEFF Research Database (Denmark)

Winkel, Anders; Borum, Jens

2009-01-01

freshwater plants with different morphology and growth characteristics (Lobelia dortmanna, Lilaeopsis macloviana, Ludwigia repens, Vallisneria americana and Hydrocotyle verticillata) are able to support photosynthesis supplied by uptake of CO2 from the sediment. Methods: Gross photosynthesis was measured......Background and Aims: Submersed plants have different strategies to overcome inorganic carbon limitation. It is generally assumed that only small rosette species (isoetids) are able to utilize the high sediment CO2 availability. The present study examined to what extent five species of submersed......, the shoot to root ratio on an areal basis was the single factor best explaining variability in the importance of sediment CO2. For Ludwigia, diffusion barriers limited uptake or transport from roots to stems and transport from stems to leaves. Conclusions: Submersed plants other than isoetids can utilize...

Aquatic weeds: their implications in Indian nuclear industry

International Nuclear Information System (INIS)

Rao, T.S.; Anup Kumar, B.; Aruna Jyothi, K.; Satpathy, K.K.

2000-01-01

The aquatic weed infestation of KAPS cooling water system, MAPS open reservoir, and the growth of filamentous algae and bacteria in the feed water unit of HWP (Kota), was investigated. The aquatic weeds identified were: Ceratophyllum, Elodea, Hydrilla verticillata, Najas and Vallisneria species. However, at HWP (Kota) filamentous alga (Nostoc punchiformis) and bacteria (Sphaerotilus natans) were found in plenty. The metabolic products when assayed in the form of total carbohydrate content released by weeds was 3.7 mg gm -1 biomass. The metabolic products adsorb on to the resin matrix and impaired its performance at HWP (K), enhanced bacterial growth on the resin beads and furthered resin deterioration. Besides, the growth of aquatic weeds also influenced the pH of the cooling water, thereby vitiating the water treatment programme. (author)

Seasonal variation in the structure of red reflectance of leaves from yellow poplar, red oak, and red maple

Science.gov (United States)

Brakke, Thomas W.; Wergin, William P.; Erbe, Eric F.; Harnden, Joann M.

1993-01-01

The light scattered from leaves was measured as a function of view angle in the principal plane for yellow poplar, red oak, and red maple. The source was a parallel-polarized helium-neon laser. Yellow poplar leaves had the highest reflectance of the three species, which may have been due to its shorter palisade cells and more extensive spongy mesophyll. Prior to senescence, there was a significant decrease, but not total extinction, in the reflectance of the beam incident at 60 deg from nadir on the adaxial side of the leaves of all three species. Low-temperature SEM observations showed differences in the surface wax patterns among the three species but did not indicate a cause of the reflectance changes other than possibly the accumulation and aging of the wax.

[Morphology, anatomy and floral biology of Cabralea canjerana (Vell.) Mart. (Meliaceae)].

Science.gov (United States)

Moscheta, Ismar S; de Souza, Luiz A; Mourão, Káthia S; da Rosa, Sônia M

2002-01-01

Cabralea canjerana (Vell.) Mart. is a tree that occurs frequently in secondary forests of Maringá, Paraná, Brazil and presents a valuable wood. Its flowering time occurs from August to October and the anthesis occurs during the night. Its flowers are visited by Lepidoptera-Noctuidae. The flowers are unisexual and solitary or arranged in panicles. The perianth presents a papillose epidermis with striate cuticle and a parenchymatic mesophyll. Ten stamens constitute the androecium and are arranged in a staminal tube with anthers. The anthers present epidermis, endothecium, two median layers and secretory tapetum with binucleate cells. The semi-inferior ovary presents anatropous, bitegmic and crassinucleate ovules. The nectaries are located in the base of the ovary and staminal tube and they present papillose epidermis with stomata and secretory parenchyma with a conspicuous phloematic tissue.

Method to reduce contamination and uptake of lead by plants from car exhaust gases

Energy Technology Data Exchange (ETDEWEB)

Isermann, K.

1977-03-01

Splashing and/or washing plants with aqueous solutions of the chelates CaEDTA (max. 5 mM/litre) or Na-polyphosphate (max. 0.5 percent) is an effective way of reducing contamination and uptake of lead by plants within regions where lead is emitted at significant levels. By chelating lead it first becomes unloaded or even negatively charged (for instance with EDTA as Pb/sub 2/EDTA or PbHEDTA/sup 1 -/ and PbEDTA/sup 2 -/). Therefore chelated lead, in contrast to normal Pb/sup 2 +/, is not absorbed, either on negatively charged surfaces of the plants (cuticula) or within the plant-tissue (mesophyll, on negatively charged cell-walls or membranes). Also, Pb-chelate has a larger diameter than bivalent Pb/sup 2 +/, resulting in a restricted lead uptake by the plant roots.

Vegetation of the eastern communal conservancies in Namibia: II. Environmental drivers

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Ben J. Strohbach

2014-11-01

Full Text Available The eastern communal conservancies are situated along the western fringe of the Kalahari basin. Under a very short rainfall gradient, the vegetation abruptly changes from microphyllous Acacia-dominated savannas to mesophyll savannas, dominated by Terminalia sericea and Combretum spp. We hypothesise that this is caused by changes in soil moisture availability brought about by changes in soil texture from loamy soils to deep sands (the ‘inverse texture effect’. For this analysis, we used vegetation and soils data derived from a recognisance survey of the natural resources of the study area. As the sites in the soil and vegetation surveys did not overlap, it was decided to use only synoptic data for the plant associations in the analysis. Non-metric multidimesional scaling ordination was utilised as ordination technique of the vegetation data and various environmental parameters, including soil texture, soil hydraulic parameters, climatic and fire regime parameters, were overlaid as biplots onto the resulting graph, as were various plant functional attributes particularly related to climatic conditions. The main environmental gradient identified within the study area is the rainfall gradient. This relatively short gradient, however, does not explain the marked change in vegetation observed within the study area. This change is attributed to the change in soil type, in particular, the soil texture and the associated soil hydraulic parameters of the soil. This gradient is closely correlated to leaf size, explaining the change from microphyll savannas to mesophyll savannas along the change from loamy to sandy soils. One of the lesser understood mechanisms for the survival of these mesophyll plants on sandy soils seems to be a deep root system, which is actively involved in water redistribution within the soil profile – by hydraulic lift, inverse hydraulic lift and stem flow. Conservation implications: Understanding these mechanisms will greatly

IRON REDUCTASE SYSTEMS ON THE PLANT PLASMA-MEMBRANE - A REVIEW

NARCIS (Netherlands)

MOOG, PR; BRUGGEMANN, W

1994-01-01

Higher plant roots, leaf mesophyll tissue, protoplasts as well as green algae are able to reduce extra-cellular ferricyanide and ferric chelates. In roots of dicotyledonous and nongraminaceous, monocotyledonous plants, the rate of ferric reduction is increased by iron deficiency. This reduction is

Factors impacting on the microbiological quality and safety of ...

African Journals Online (AJOL)

hope&shola

2010-12-06

Dec 6, 2010 ... microbiological quality and safety of processed hake. Samples were collected along the processing line; the general microbiological quality (mesophylic and psychrotrophic aerobic plate counts), total. Vibrio species and common fish spoilage bacterial counts were performed. The results constantly showed ...

How active ingredient localisation in plant tissues determines the targeted pest spectrum of different chemistries

DEFF Research Database (Denmark)

Buchholz, Anke; Trapp, Stefan

2016-01-01

information sets revealed that the intracellular localisation of active ingredients determines the performance of test compounds against different target pests because of different feeding behaviours: mites feed on mesophyll, and aphids and whiteflies mostly in the vascular system. Polar compounds have a slow...

Ascorbic acid is a key participant during the interactions between ...

Indian Academy of Sciences (India)

AOX pathway as a percentage of the total respiration in Arabidopsis mesophyll ... synthesis catalysed by L-GalLDH, is located on the inner ... −1. ) and 16 h dark photoperiod and a temperature of 22–23°C. Nutrient solution (Somerville.

Photosynthetic metabolism of malate and aspartate in Flaveria trinervia a C4 dicot

International Nuclear Information System (INIS)

Moore, B.A.

1986-01-01

C 4 species are known to vary in their apparent relative use of malate and aspartate to mediate carbon flux through the C 4 cycle. These studies investigate some of the adjustments in photosynthetic carbon metabolism that occur during a dark to light transition and during expansion of leaves of Flaveria trinervia, a C 4 dicot. Enzyme localization studies with isolated leaf mesophyll and bundle sheath protoplasts, indicated that both C 4 acids are formed in the mesophyll chloroplast, and that aspartate is metabolized to malate in the bundle sheath chloroplast prior to decaroxylation there. During photosynthetic induction, the partitioning of 14 CO 2 between malate and aspartate showed a single oscillation of increased aspartate labelling after 5 min of illumination. Turnover of [4-14C] (malate plus aspartate) was slow initially during illumination, prior to establishment of active pools of C 4 cycle metabolites

Photosynthesis Decrease and Stomatal Control of Gas Exchange in Abies alba Mill. in Response to Vapor Pressure Difference.

Science.gov (United States)

Guehl, J M; Aussenac, G

1987-02-01

The responses of steady state CO(2) assimilation rate (A), transpiration rate (E), and stomatal conductance (g(s)) to changes in leaf-to-air vapor pressure difference (DeltaW) were examined on different dates in shoots from Abies alba trees growing outside. In Ecouves, a provenance representative of wet oceanic conditions in Northern France, both A and g(s) decreased when DeltaW was increased from 4.6 to 14.5 Pa KPa(-1). In Nebias, which represented the dry end of the natural range of A. alba in southern France, A and g(s) decreased only after reaching peak levels at 9.0 and 7.0 Pa KPa(-1), respectively. The representation of the data in assimilation rate (A) versus intercellular CO(2) partial pressure (C(i)) graphs allowed us to determine how stomata and mesophyll photosynthesis interacted when DeltaW was increased. Changes in A were primarily due to alterations in mesophyll photosynthesis. At high DeltaW, and especially in Ecouves when soil water deficit prevailed, A declined, while C(i) remained approximately constant, which may be interpreted as an adjustment of g(s) to changes in mesophyll photosynthesis. Such a stomatal control of gas exchange appeared as an alternative to the classical feedforward interpretation of E versus DeltaW responses with a peak rate of E. The gas exchange response to DeltaW was also characterized by considerable deviations from the optimization theory of IR Cowan and GD Farquhar (1977 Symp Soc Exp Biol 31: 471-505).

Implementing a Probabilistic Line of Sight in EASEE (Environmental Awareness for Sensor and Emitter Employment)

Science.gov (United States)

2015-05-01

prefera- ble to other options with limited coverage extent, such as the National Land Cover Database . Midway through the project, the Army Corps...and mesophyll, which support photosynthesis . The growth of new leaves and the dropping of leaves are key events for visibility considerations; each

Characterization of natural leaf senescence in tobacco (Nicotiana tabacum) plants grown in vitro

Czech Academy of Sciences Publication Activity Database

Uzelac, B.; Janošević, D.; Simonović, A.; Motyka, Václav; Dobrev, Petre; Budimir, S.

2016-01-01

Roč. 253, č. 2 (2016), s. 259-275 ISSN 0033-183X R&D Projects: GA ČR(CZ) GAP506/11/0774 Institutional support: RVO:61389030 Keywords : Leaf senescence * Mesophyll ultrastructure * Phytohormones Subject RIV: EF - Botanics Impact factor: 2.870, year: 2016

Superimposed behaviour of g(m) under ABA-induced stomata closing and low CO2

Czech Academy of Sciences Publication Activity Database

Tazoe, Y.; Šantrůček, Jiří

2015-01-01

Roč. 38, č. 3 (2015), s. 385-387 ISSN 0140-7791 R&D Projects: GA ČR(CZ) GAP501/12/1261 Institutional support: RVO:60077344 Keywords : Mesophyll conductance * Carbon-isotope discrimination * Photosnthesis * Plants Subject RIV: CE - Biochemistry Impact factor: 6.169, year: 2015

Mesoporous silica nanoparticles as a biomolecule delivery vehicle in plants

Energy Technology Data Exchange (ETDEWEB)

Hussain, Hashmath I., E-mail: hashmath.i@deakin.edu.au [Deakin University, Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences (Australia); Yi, Zhifeng [Deakin University, Institute for Frontier Materials (Australia); Rookes, James E. [Deakin University, Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences (Australia); Kong, Lingxue X. [Deakin University, Institute for Frontier Materials (Australia); Cahill, David M. [Deakin University, Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences (Australia)

2013-06-15

We report the uptake by wheat, lupin and Arabidopsis of mesoporous silica nanoparticles functionalised with amine cross-linked fluorescein isothiocyanate (MSN-APTES-FITC). The preparation of these particles at room temperature enabled the synthesis of 20 nm particles that contained a network of interconnected pores around 2 nm in diameter. The uptake and distribution of these nanoparticles were examined during seed germination, in roots of plants grown in a hydroponic system and in whole leaves and roots of plants via vacuum infiltration. The nanoparticles did not affect seed germination in lupin and there was no phytotoxicity. Following germination of wheat and lupin grown in a nutrient solution containing nanoparticles, they were found within cells and cell walls of the emerging root and in the vascular transport elements, the xylem, and in other associated cells. In leaves and roots of Arabidopsis the nanoparticles were found, following vacuum infiltration of whole seedlings, to be taken up by the entire leaf and they were principally found in the intercellular spaces of the mesophyll but also throughout much of the root system. We propose that MSNs could be used as a novel delivery system for small molecules in plants.

Leaf and inflorescence axis anatomy of Brazilian species of Rapateoideae (Rapateaceae, Poales

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Ângela L. Daltin

2015-03-01

Full Text Available The anatomy of leaves and inflorescence axes of Spathanthus (2 spp., Rapatea (2 spp., Cephalostemon(1 sp., and Duckea(1 sp. (Rapateoideae, Rapateaceae was studied to identify useful characters for taxonomy. The cross-section shape of inflorescence axis differentiates the genera, while the cross-section shape and structure of leaf midrib has a specific value. The following characteristics are exclusive of Spathanthus: silica cells randomly distributed in the leaf epidermis; plicate chlorenchyma in the leaf blade; presence of fiber bundles in the mesophyll and in the inflorescence axis parenchyma. Spathanthus is also distinguished by the number, type and distribution of vascular bundles in the inflorescence axis. The genus Rapatea is characterized by the presence of stomata and silica cells only on the abaxial epidermis of the leaves and chlorenchyma composed of arm cells in the leaf blade. Characteristics with diagnostic value for Cephalostemon riedelianusare: leaf epidermal cells with straight to slightly sinuous walls in frontal view, inflorescence axes presenting a defined cortex, fiber bundles facing the larger vascular bundles and a fistulous pith. The anatomical characteristics of the leaves and inflorescence axes thus proved to be of taxonomic value in generic and specific levels. They are also useful to differentiate Rapateoideae from other subfamilies of Rapateaceae.

Variations in the dorso-ventral organization of leaf structure and Kranz anatomy coordinate the control of photosynthesis and associated signalling at the whole leaf level in monocotyledonous species.

Science.gov (United States)

Soares-Cordeiro, Ana Sofia; Driscoll, Simon P; Pellny, Till K; Olmos, Enrique; Arrabaça, Maria Celeste; Foyer, Christine H

2009-12-01

Photosynthesis and associated signalling are influenced by the dorso-ventral properties of leaves. The degree of adaxial/abaxial symmetry in stomatal numbers, photosynthetic regulation with respect to light orientation and the total section areas of the bundle sheath (BS) cells and the surrounding mesophyll (M) cells on the adaxial and abaxial sides of the vascular bundles were compared in two C(4)[Zea mays (maize) and Paspalum dilatatum] and one C(3)[Triticum turgidum (Durum wheat)] monocotyledonous species. The C(3) leaves had a higher degree of dorso-ventral symmetry than the C(4) leaves. Photosynthetic regulation was the same on each side of the wheat leaves, as were stomatal numbers and the section area of the BS relative to that of the M cells (BS/M section area ratio). In contrast, photosynthetic regulation in maize and P. dilatatum leaves showed a marked surface-specific response to light orientation. Compared to the adaxial sides of the C(4) monocotyledonous leaves, the abaxial surfaces had more stomata and the BS/M section area ratio was significantly higher. Differences in dorso-ventral structure, particularly in Kranz anatomy, serve not only to maximize photosynthetic capacity with respect light orientation in C(4) monocotyledonous leaves but also allow adaxial and abaxial-specific signalling from the respective M cells.

Variability of anatomical-physiological traits in black locust clones - Robinia pseudoacacia L

Directory of Open Access Journals (Sweden)

Orlović Saša S.

2004-01-01

Full Text Available Variability within R. pseudoacacia species represents an important factor in selection of fast-growing genotypes. Therefore, it is important to identify superior individuals according to their anatomical and physiological traits. This paper presents the results of a study of genotype variability of the main leaf anatomical (frequency, length and width of stomata, leaflet thickness among veins, leaflet thickness on the main vein, mesophyll thickness, length and width of vascular bundle of main vein and physiological (leaf area, photosynthetic pigments content and content of N P, K, Ca, Na parameters among five clones of Robinia pseudoacacia L. Significant interclonal variations were observed in the investigated parameters. Clone R-56 had the highest N, P, and K concentrations, the largest mesophyll volume and the highest pigment concentration. We concluded that the clone R-56, although without a remarkable leaf area, possesses the ability for high photosynthetic production. The results are going to be used in further work on selection.

FOLIAR ANATOMY OF ENDEMICS SPECIES OF Cattleya (ORCHIDACEAE ENDEMIC FROM GUIANA SHIELD

Directory of Open Access Journals (Sweden)

Graciene Tomaz Carneiro

2017-03-01

Full Text Available The main goal of this study was characterize the leaf's anatomical aspects of Cattleya jenmanii Rolfe and e C. lawrenceana Rchb. f., describing its anatomical structures in order to increase the knowledge of this endemic species from the region of the Guiana Shield. Besides, it also intended to identify foliar characters to assist in the anatomical comparison of these species. For anatomical study, the material was fixed in FAA and to make the slides we used the usual cut freehand technique and stained with double staining from Safranin with Blue Astra (Safrablau. C. jenmanii and C. lawrenceana has fleshy leaves covered with a thick cuticle. The mesophyll presented dorsiventral with collateral vascular bundles. A large number of bundles of smaller caliber fibers are distributed in the mesophyll poles. Only the presence of a subepidermal layer of fibers differed C. lawrenceana from C. jenmanii. Keyword: Roraima; Guiana Shield; Cattleya; Amazon Basin.

Hg localisation in Tillandsia usneoides L. (Bromeliaceae), an atmospheric biomonitor

Energy Technology Data Exchange (ETDEWEB)

Filho, G.M.A. [Instituto de Pesquisas Jardim Botanico do Rio de Janeiro (Brazil). Programa Zona Costeira; Andrade, L.R.; Farina, M. [Cidade Universitaria, Rio de Janeiro (Brazil). Instituto de Ciencias Biomedicas, Departamento de Anatomia; Malm, O. [Cidade Universitaria, Rio de Janeiro (Brazil). Instituto de Biofisica Carlos Chagas Filho, Laboratorio de Radioisotopos Eduardo Penna Franca

2002-07-01

The Spanish moss, Tillandsia usneoides, has been applied as an atmospheric biomonitor of Hg contamination, although the mechanism of metal plant accumulation has not been understood until now. In the present work, analytical scanning electron microscopy (SEM) was used to localize Hg in T. usneoides exposed to a Hg-air-contaminated area during 15 days. After this period, Hg was determined by the flow injection mercury system, and plants were prepared for SEM observation and energy-dispersive X-ray analysis. A concentration of 2702{+-}318{mu}g Hgg{sup -1} was determined in exposed plants. The presented microanalytical results demonstrated that Hg was partly associated with atmospheric particles deposited upon the plant surface, but it was highly absorbed by the scales, stem and leaves surfaces and less absorbed by epidermal cells of T. usneoides. No Hg was detected in mesophyll parenchyma or in vascular system cells. The great surface adsorption area provided by the scales, in addition to the characteristics of T. usneoides morphology, especially of the node region, are suggested to confer the great capability of T. usneoides in Hg holding. (author)

Hg localisation in Tillandsia usneoides L. (Bromeliaceae), an atmospheric biomonitor

Science.gov (United States)

Amado Filho, G. M.; Andrade, L. R.; Farina, M.; Malm, O.

The Spanish moss, Tillandsia usneoides, has been applied as an atmospheric biomonitor of Hg contamination, although the mechanism of metal plant accumulation has not been understood until now. In the present work, analytical scanning electron microscopy (SEM) was used to localize Hg in T. usneoides exposed to a Hg-air-contaminated area during 15 days. After this period, Hg was determined by the flow injection mercury system, and plants were prepared for SEM observation and energy-dispersive X-ray analysis. A concentration of 2702±318 μg Hg g -1 was determined in exposed plants. The presented microanalytical results demonstrated that Hg was partly associated with atmospheric particles deposited upon the plant surface, but it was highly absorbed by the scales, stem and leaves surfaces and less absorbed by epidermal cells of T. usneoides. No Hg was detected in mesophyll parenchyma or in vascular system cells. The great surface adsorption area provided by the scales, in addition to the characteristics of T. usneoides morphology, especially of the node region, are suggested to confer the great capability of T. usneoides in Hg holding.

Studies on the treatment efficiency of sediment phosphorus with a combined technology of PCFM and submerged macrophytes.

Science.gov (United States)

Zhang, Yi; He, Feng; Xia, Shibin; Zhou, Qiaohong; Wu, Zhenbin

2015-11-01

The removal efficiency of sediment phosphorus (P) in all fractions with a combined technology of porous ceramic filter media (PCFM) and submerged macrophytes was studied in Donghu Lake, Wuhan, China. The adsorption kinetic models of the sediment P in all fractions on PCFM could be described well by a power function equations (Qt = k · t(a), 0 macrophyte, was higher for all P forms than that of the combination of PCFM and another macrophyte, Vallisneria spiralis. This study suggested that the combination of PCFM and macrophytes could achieve a synergetic sediment P removal because the removal rates of the combinations were higher than the sum of that of PCFM and macrophytes used separately. The combined technology could be further applied to treat internal P loading in eutrophic waters. Copyright © 2015 Elsevier Ltd. All rights reserved.

Variation in the number of nucleoli and incomplete homogenization of 18S ribosomal DNA sequences in leaf cells of the cultivated Oriental ginseng (Panax ginseng Meyer).

Science.gov (United States)

Chelomina, Galina N; Rozhkovan, Konstantin V; Voronova, Anastasia N; Burundukova, Olga L; Muzarok, Tamara I; Zhuravlev, Yuri N

2016-04-01

Wild ginseng, Panax ginseng Meyer, is an endangered species of medicinal plants. In the present study, we analyzed variations within the ribosomal DNA (rDNA) cluster to gain insight into the genetic diversity of the Oriental ginseng, P. ginseng, at artificial plant cultivation. The roots of wild P. ginseng plants were sampled from a nonprotected natural population of the Russian Far East. The slides were prepared from leaf tissues using the squash technique for cytogenetic analysis. The 18S rDNA sequences were cloned and sequenced. The distribution of nucleotide diversity, recombination events, and interspecific phylogenies for the total 18S rDNA sequence data set was also examined. In mesophyll cells, mononucleolar nuclei were estimated to be dominant (75.7%), while the remaining nuclei contained two to four nucleoli. Among the analyzed 18S rDNA clones, 20% were identical to the 18S rDNA sequence of P. ginseng from Japan, and other clones differed in one to six substitutions. The nucleotide polymorphism was more expressed at the positions 440-640 bp, and distributed in variable regions, expansion segments, and conservative elements of core structure. The phylogenetic analysis confirmed conspecificity of ginseng plants cultivated in different regions, with two fixed mutations between P. ginseng and other species. This study identified the evidences of the intragenomic nucleotide polymorphism in the 18S rDNA sequences of P. ginseng. These data suggest that, in cultivated plants, the observed genome instability may influence the synthesis of biologically active compounds, which are widely used in traditional medicine.

Accumulation of defence-related transcripts and cloning of a chitinase mRNA from pea leaves (Pisum sativum L.) inoculated with Ascochyta pisi Lib

DEFF Research Database (Denmark)

Vad, Knud; de Neergaard, Eigil; Madriz-Ordeñana, Kenneth

1993-01-01

The race specific resistance of pea to Ascochyta pisi Lib. was shown to be exhibited as a hypersensitive response associated with the production of polyphenolic substances in epidermal and mesophyll cells. The levels of transcripts representing a pathogenesis-related (PR) protein (chitinase......) and an enzyme of phytoalexin biosynthesis (chalcone synthase) were shown to accumulate more rapidly during the hypersensitive response than during lesion development in the compatible interaction. A full-length (1143 bp) cDNA sequence of a pea chitinase (EC 3.2.1.14) (coding for an approx. 34 500 Da protein......) was deduced by combining the overlapping sequences of three clones obtained following PCR amplification of cDNA prepared from mRNA isolated 24 h after inoculation of pea leaves with Ascochyta pisi. The combined sequences were identified as a class I chitinase corresponding to the basic A1-chitinase enzyme...

Comparative Leaf Anatomy of Gaiadendron punctatum and Tripodanthus belmirensis (Loranthaceae

Directory of Open Access Journals (Sweden)

Isabel Carmona Gallego

2018-01-01

Full Text Available Leaf anatomy of Gaiadendron punctatum and Tripodanthus belmirensis species is described with the aim of exploring possible characters that allow a precise identification of these two genera, characterized by tree or shrub habit, belonging to Loranthaceae family. Samples were processed and stained with routine techniques for observation on optical microscope. Both species showed similarities, such as one-layered epidermis and rubiaceous type stomata. However, a different anatomic composition was observed in aspects such as: mesophyll, position and shape of epidermic cells and presence of suber accumulations in G. punctatum, or numerous idioblasts in T. belmirensis. A dichotomous key was constructed for species determination based on anatomical leaf characters, between the species of genus Tripodanthus and G. punctatum. Furthermore, the use of anatomical characters in determination and support of distinguishable taxonomical entities inside Loranthaceae is also briefly discussed.

Zelltyp-spezifische Mikroanalyse von Arabidopsis thaliana-Blättern

Science.gov (United States)

Brandt, Stephan Peter

2002-04-01

specificity of gene activity some results were reconsidered using tissue specific northern blot hybridisations and real time RT-PCR, respectively. Secondly, metabolites (including inorganic ions) were investigated: Because gas chromatography-mass spectrometry does not reveal the sensitivity which in necessary for the analysis of even multiple pooled single cell samples capillary electrophoresis was applied for these studies. This method has a high potential as it needs only small amounts of starting material, has uncomparable low detection limits and exhibits a high number of theoretical plates. The analysis of inorganic anions and carbohydrates needs further optimisations. Using UV absorption-detection potassium could be detected in different cell types whereas the concentrations in mesophyll and epidermis were found around 25 mM each. These concentrations are lower than in other species as Solanum tuberosum or Hordeum vulgare. For investigations of amino acids the cell samples were derivatized to make the use of laser induced fluorescence-detection capable. In samples derived from pumpkin (Cucurbita maxima) mesophyll twelve amino acids could be detected and identified. The transfer of this method to A. thaliana derived samples exhibited no results which may be due to the low concentration of free amino acids in these plants. Finally, a method was developed with which the existence of known and unknown proteins in tissue specific samples could be monitored. For this, mechanical micro dissection was used to: After embedding and sectioning the tissue of interest was cut out by an vibrating steel chisel to get homogenous samples. The proteins contained in these tissue pieces were extracted and separated by one dimensional SDS polyacrylamid gel electrophoresis. Several protein bands could be detected after staining with either silver or coomassie blue stain. These bands were cut out and sequenced by mass spectrometry. The large subunit of rubisco as well as one chlorophyll

Vegetative anatomical adaptations of Epidendrum radicans (Epidendroideae, Orchidaceae to epiphytic conditions of growth

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Muthukumar Thangavelu

2017-12-01

Full Text Available The anatomical properties of leaf, stem, and root of Epidendrum radicans Pav. ex Lindl., belonging to the subfamily Epidendroideae (Orchidaceae were investigated for adaptations to stressed habitats. The anatomical investigation revealed that leaves of E. radicans have a thick cuticle (3–4 µm and paracytic type of stomata. Foliar epidermal cells are conical on the adaxial surface and rectangular in the abaxial surface, distinct hypodermis absent, and uniseriate fiber bundles are arranged in both sides of the leaves. The foliar mesophyll is homogenous and starch grains and raphides present. The leaf sheath covering the stem have cuticle restricted to the outer surface and air spaces are present. The stem has a cuticulerized uniseriate epidermis and a uniseriate hypodermis. The cortex and a parenchymatous ground tissue of the stem are separated by a layer of sclerenchymatous band. Vascular bundles are collateral and their size generally increases from the periphery towards the center. A sclerenchymatous patch covers the phloem pole, whereas the xylem is covered by thin-walled parenchymatous cells. The roots possess Epidendrum-type velamen. Cover cells present. Uniseriate dimorphic exodermis consists of U-thickened long cells and thin-walled passage cells. The endodermal cells O-thickened, pericycle sclerenchymatous, xylem 10–14 arched. The pith is sclerenchymatous, but parenchymatous at the center. The anatomical examination of E. radicans revealed adaptations to moisture stress conditions like thick cuticle covering the leaves and stem, water storage cells, multilayered velamen and dimorphic exodermis.

Hydrogen sulphide improves adaptation of Zea mays seedlings to iron deficiency.

Science.gov (United States)

Chen, Juan; Wu, Fei-Hua; Shang, Yu-Ting; Wang, Wen-Hua; Hu, Wen-Jun; Simon, Martin; Liu, Xiang; Shangguan, Zhou-Ping; Zheng, Hai-Lei

2015-11-01

Hydrogen sulphide (H2S) is emerging as a potential molecule involved in physiological regulation in plants. However, whether H2S regulates iron-shortage responses in plants is largely unknown. Here, the role of H2S in modulating iron availability in maize (Zea mays L. cv Canner) seedlings grown in iron-deficient culture solution is reported. The main results are as follows: Firstly, NaHS, a donor of H2S, completely prevented leaf interveinal chlorosis in maize seedlings grown in iron-deficient culture solution. Secondly, electron micrographs of mesophyll cells from iron-deficient maize seedlings revealed plastids with few photosynthetic lamellae and rudimentary grana. On the contrary, mesophyll chloroplasts appeared completely developed in H2S-treated maize seedlings. Thirdly, H2S treatment increased iron accumulation in maize seedlings by changing the expression levels of iron homeostasis- and sulphur metabolism-related genes. Fourthly, phytosiderophore (PS) accumulation and secretion were enhanced by H2S treatment in seedlings grown in iron-deficient solution. Indeed, the gene expression of ferric-phytosiderophore transporter (ZmYS1) was specifically induced by iron deficiency in maize leaves and roots, whereas their abundance was decreased by NaHS treatment. Lastly, H2S significantly enhanced photosynthesis through promoting the protein expression of ribulose-1,5-bisphosphate carboxylase large subunit (RuBISCO LSU) and phosphoenolpyruvate carboxylase (PEPC) and the expression of genes encoding RuBISCO large subunit (RBCL), small subunit (RBCS), D1 protein (psbA), and PEPC in maize seedlings grown in iron-deficient solution. These results indicate that H2S is closely related to iron uptake, transport, and accumulation, and consequently increases chlorophyll biosynthesis, chloroplast development, and photosynthesis in plants. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Foliar Nutrient Distribution Patterns in Sympatric Maple Species Reflect Contrasting Sensitivity to Excess Manganese.

Science.gov (United States)

Fernando, Denise R; Marshall, Alan T; Lynch, Jonathan P

2016-01-01

Sugar maple and red maple are closely-related co-occurring tree species significant to the North American forest biome. Plant abiotic stress effects including nutritional imbalance and manganese (Mn) toxicity are well documented within this system, and are implicated in enhanced susceptibility to biotic stresses such as insect attack. Both tree species are known to overaccumulate foliar manganese (Mn) when growing on unbuffered acidified soils, however, sugar maple is Mn-sensitive, while red maple is not. Currently there is no knowledge about the cellular sequestration of Mn and other nutrients in these two species. Here, electron-probe x-ray microanalysis was employed to examine cellular and sub-cellular deposition of excessively accumulated foliar Mn and other mineral nutrients in vivo. For both species, excess foliar Mn was deposited in symplastic cellular compartments. There were striking between-species differences in Mn, magnesium (Mg), sulphur (S) and calcium (Ca) distribution patterns. Unusually, Mn was highly co-localised with Mg in mesophyll cells of red maple only. The known sensitivity of sugar maple to excess Mn is likely linked to Mg deficiency in the leaf mesophyll. There was strong evidence that Mn toxicity in sugar maple is primarily a symplastic process. For each species, leaf-surface damage due to biotic stress including insect herbivory was compared between sites with acidified and non-acidified soils. Although it was greatest overall in red maple, there was no difference in biotic stress damage to red maple leaves between acidified and non-acidified soils. Sugar maple trees on buffered non-acidified soil were less damaged by biotic stress compared to those on unbuffered acidified soil, where they are also affected by Mn toxicity abiotic stress. This study concluded that foliar nutrient distribution in symplastic compartments is a determinant of Mn sensitivity, and that Mn stress hinders plant resistance to biotic stress.

Anatomical features of pepper plants (Capsicum annuum L.) grown under red light-emitting diodes supplemented with blue or far-red light

Science.gov (United States)

Schuerger, A. C.; Brown, C. S.; Stryjewski, E. C.

1997-01-01

Pepper plants (Capsicum annuum L. cv., Hungarian Wax) were grown under metal halide (MH) lamps or light-emitting diode (LED) arrays with different spectra to determine the effects of light quality on plant anatomy of leaves and stems. One LED (660) array supplied 90% red light at 660 nm (25nm band-width at half-peak height) and 1% far-red light between 700-800nm. A second LED (660/735) array supplied 83% red light at 660nm and 17% far-red light at 735nm (25nm band-width at half-peak height). A third LED (660/blue) array supplied 98% red light at 660nm, 1% blue light between 350-550nm, and 1% far-red light between 700-800nm. Control plants were grown under broad spectrum metal halide lamps. Plants were gron at a mean photon flux (300-800nm) of 330 micromol m-2 s-1 under a 12 h day-night photoperiod. Significant anatomical changes in stem and leaf morphologies were observed in plants grown under the LED arrays compared to plants grown under the broad-spectrum MH lamp. Cross-sectional areas of pepper stems, thickness of secondary xylem, numbers of intraxylary phloem bundles in the periphery of stem pith tissues, leaf thickness, numbers of choloplasts per palisade mesophyll cell, and thickness of palisade and spongy mesophyll tissues were greatest in peppers grown under MH lamps, intermediate in plants grown under the 660/blue LED array, and lowest in peppers grown under the 660 or 660/735 LED arrays. Most anatomical features of pepper stems and leaves were similar among plants grown under 660 or 660/735 LED arrays. The effects of spectral quality on anatomical changes in stem and leaf tissues of peppers generally correlate to the amount of blue light present in the primary light source.

Anatomia e histoquímica das folhas de Senna alata Anatomy and histochemistry of Senna alata leaves

Directory of Open Access Journals (Sweden)

I.M.C. Rodrigues

2009-01-01

environments. Also, the leaves are rich in calcium oxalate crystals along their veins, a characteristic of Caesalpiniodeae subfamily. Phenolic compounds, such as flavonoids and anthraquinones, were found in epidermal and basal trichome cells, and scattered cells from palisade parenchyma, especially near the midvein. This study confirms the presence of known allelochemical classes in different types of cells in Senna alata mesophyll.

Quantitative imaging of radial oxygen loss from Valisneria spiralis roots with a fluorescent planar optode

International Nuclear Information System (INIS)

Han, Chao; Ren, Jinghua; Tang, Hao; Xu, Di; Xie, Xianchuan

2016-01-01

Oxygen (O_2) availability within the sediment–root interface is critical to the survival of macrophytes in O_2-deficient sediment; however, our knowledge of the fine-scale impact of macrophyte roots upon the spatiotemporal dynamics of O_2 is relatively limited. In this study, a non-invasive imaging technology was utilized to map O_2 micro-distribution around Vallisneria spiralis. Long-term imaging results gathered during a 36 day-period revealed an abundance of O_2 spatiotemporal patterns ranging from 0 to 250 μmol L"− "1. The root-induced O_2 leakage and consequent oxygenated area were stronger in the vicinity of the basal root compared to that found in the root tip. The O_2 images revealed V. spiralis exhibited radial O_2 loss (ROL) along the entire root, and the O_2 distribution along the root length showed a high degree of small-scale spatial heterogeneity decreasing from 80% at the basal root surface to 10% at the root tip. The oxygenated zone area around the roots increased as O_2 levels increased with root growth and irradiance intensities ranging from 0 to 216 μmol photons m"− "2 s"− "1. A weak ROL measuring < 20% air saturation around the basal root surface was maintained in darkness, which was presumably attributed to the O_2 supply from overlying water via plant aerenchyma. The estimated total O_2 release to the rhizosphere of V. spiralis was determined to range from 8.80 ± 7.32 to 30.34 ± 17.71 nmol m"− "2 s"− "1, which is much higher than many other macrophyte species. This O_2 release may be an important contribution to the high-capacity of V. spiralis for quickly colonizing anaerobic sediment. - Highlights: • Planar imaging method was used to map O2 micro-distribution. • Highly dynamic rhizospheric O2-spatiotemporal distribution was observed. • O_2 leakage along the entire root of Vallisneria spirals were defined. • The ROL rates of 8.80–30.34 nmol m"− "2 s"− "1 were measured over a 36-day growth. • ROL was closely

Significance and transmission of maize streak virus disease in Africa ...

African Journals Online (AJOL)

STORAGESEVER

2008-12-29

Dec 29, 2008 ... soil nutrients, altitude and temperature on the biology of maize streak virus (MSV) / vector populations is discussed. ... status of maize host plants and its effects on population dynamics of Cicadulina mbila Naudé. (Homoptera: ..... time necessary for the leafhopper to reach the mesophyll of the leaf and ingest ...

Are changes in sulfate assimilation pathway needed for evolution of C4 photosynthesis?

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Silke Christine Weckopp

2015-01-01

Full Text Available C4 photosynthesis characteristically features a cell-specific localization of enzymes involved in CO2 assimilation in bundle sheath cells or mesophyll cells. Interestingly, enzymes of sulfur assimilation are also specifically present in bundle sheath cells of maize and many other C4 species. This localization, however, could not be confirmed in C4 species of the genus Flaveria. It was, therefore, concluded that the bundle sheath localization of sulfate assimilation occurs only in C4 monocots. However, recently the sulfate assimilation pathway was found coordinately enriched in bundle sheath cells of Arabidopsis, opening new questions about the significance of such cell-specific localization of the pathway. In addition, next generation sequencing revealed expression gradients of many genes from C3 to C4 species and mathematical modelling proposed a sequence of adaptations during the evolutionary path from C3 to C4. Indeed, such gradient, with higher expression of genes for sulfate reduction in C4 species, has been observed within the genus Flaveria. These new tools provide the basis for reexamining the intriguing question of compartmentalization of sulfur assimilation. Therefore, this review summarizes the findings on spatial separation of sulfur assimilation in C4 plants and Arabidopsis, assesses the information on sulfur assimilation provided by the recent transcriptomics data and discusses their possible impact on understanding this interesting feature of plant sulfur metabolism to find out whether changes in sulfate assimilation are part of a general evolutionary trajectory towards C4 photosynthesis.

Inhibition of loading of 14C assimilate by p-chloromercuribenzenesulfonic acid

International Nuclear Information System (INIS)

Bourquin, S.; Bonnemain, J.L.; Delrot, S.

1990-01-01

The apoplast of mature leaves excised from broadbean (Vicia faba L.) plants was infiltrated with 2 millimolar p-chloromercuribenzenesulfonic acid (PCMBS) via the transpiration stream, and the ability of the tissues to take up sugars was tested. An infiltration time of 75 minutes was sufficient of obtain a maximal (75%) inhibition of exogenous [ 14 C]sucrose (1 millimolar) uptake. This infiltration affected neither CO 2 assimilation nor the transmembrane potential difference of leaf cells but strongly inhibited phloem loading of endogenous [ 14 C] assimilates. The study of the symplastic relations between the different cell types of the mature leaf showed that the density of the plasmodesmata is generally very low in comparison with other species investigated so far, particularly when considering the mesophyll/bundle sheath and the bundle sheath/phloem cells connections, as well as the connections of the transfer cell-sieve tube complex with the surrounding cells. These three successive barriers therefore strongly limit the possibilities of symplastic transit of the assimilates to the conducting cells. The comparison of the densities of plasmodesmata in an importing and an exporting leaf suggests that the maturation of the leaf is characterized by a marked symplastic isolation of the phloem, and, within the phloem itself, by the isolation of the conducting complex. As a consequence, these physiological and cytological data demonstrate the apoplastic nature of loading in the mature leaf of Vicia faba, this species undoubtedly presenting a typical model for apoplastic loading

Comparative conventional- and quantum dot-labelling strategies for LPS binding site detection in Arabidopsis thaliana mesophyll protoplasts

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Londiwe Siphephise Mgcina

2015-05-01

Full Text Available Lipopolysaccharide (LPS from Gram-negative bacteria is recognized as a microbe-associated molecular pattern (MAMP and not only induces an innate immune response in plants, but also stimulates the development of characteristic defense responses. However, identification and characterization of a cell surface LPS-receptor/binding site, as described in mammals, remains elusive in plants. As an amphiphilic, macromolecular lipoglycan, intact LPS potentially contains three MAMP-active regions, represented by the O-polysaccharide chain, the core and the lipid A. Binding site studies with intact labelled LPS were conducted in Arabidopsis thaliana protoplasts and quantified using flow cytometry fluorescence changes. Qdots, which allow non-covalent, hydrophobic labelling were used as a novel strategy in this study and compared to covalent, hydrophilic labelling with Alexa 488. Affinity for LPS-binding sites was clearly demonstrated by concentration-, temperature- and time-dependent increases in protoplast fluorescence following treatment with the labelled LPS. Moreover, this induced fluorescence increase was convincingly reduced following pre-treatment with excess unlabeled LPS, thereby indicating reversibility of LPS binding. Inhibition of the binding process is also reported using endo- and exocytosis inhibitors. Here, we present evidence for the anticipated presence of LPS-specific binding sites in Arabidopsis protoplasts, and furthermore propose Qdots as a more sensitive LPS-labelling strategy in comparison to the conventional Alexa 488 hydrazide label for binding studies.

Effects of ozone on the foliar histology of the mastic plant (Pistacia lentiscus L.)

International Nuclear Information System (INIS)

Reig-Arminana, J.; Calatayud, V.; Cervero, J.; Garcia-Breijo, F.J.; Ibars, A.; Sanz, M.J.

2004-01-01

An open-top chamber study was conducted to investigate the tissue and cellular-level foliar effects of ozone (O 3 ) on a Mediterranean evergreen species, the mastic plant (Pistacia lentiscus L.). Plants were exposed at three different O 3 levels, and leaf samples were collected periodically from the beginning of the exposure. Although no visible foliar injury was evident, alterations of the plastids and vacuoles in the mesophyll were observed. Senescence processes were accelerated with an anomalous stacking of tannin vacuoles, and a reduction in the size and number of the chloroplasts. Overall, most of the modifications induced by O 3 were consistent with previously reported observations on deciduous broadleaf species, with the exception of alterations in the cells covering the secretory channels, reported here as a new finding. Comments on the feasibility of using microscopy to validate O 3 related field observations and subtle foliar injury are also given

Evaluation of the potential of Pistia stratiotes L. (water lettuce for bioindication and phytoremediation of aquatic environments contaminated with arsenic

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FS Farnese

Full Text Available Specimens of Pistia stratiotes were subjected to five concentrations of arsenic (As for seven days. Growth, As absorption, malondialdehyde (MDA content, photosynthetic pigments, enzymatic activities, amino acids content and anatomical changes were assessed. Plant arsenic accumulation increased with increasing metalloid in the solution, while growth rate and photosynthetic pigment content decreased. The MDA content increased, indicating oxidative stress. Enzymatic activity and amino acids content increased at the lower doses of As, subsequently declining in the higher concentrations. Chlorosis and necrosis were observed in the leaves. Leaves showed starch accumulation and increased thickness of the mesophyll. In the root system, there was a loss and darkening of roots. Cell layers formed at the insertion points on the root stems may have been responsible for the loss of roots. These results indicate that water lettuce shows potential for bioindication and phytoremediation of As-contaminated aquatic environments.

Evaluation of the potential of Pistia stratiotes L. (water lettuce) for bioindication and phytoremediation of aquatic environments contaminated with arsenic.

Science.gov (United States)

Farnese, F S; Oliveira, J A; Lima, F S; Leão, G A; Gusman, G S; Silva, L C

2014-08-01

Specimens of Pistia stratiotes were subjected to five concentrations of arsenic (As) for seven days. Growth, As absorption, malondialdehyde (MDA) content, photosynthetic pigments, enzymatic activities, amino acids content and anatomical changes were assessed. Plant arsenic accumulation increased with increasing metalloid in the solution, while growth rate and photosynthetic pigment content decreased. The MDA content increased, indicating oxidative stress. Enzymatic activity and amino acids content increased at the lower doses of As, subsequently declining in the higher concentrations. Chlorosis and necrosis were observed in the leaves. Leaves showed starch accumulation and increased thickness of the mesophyll. In the root system, there was a loss and darkening of roots. Cell layers formed at the insertion points on the root stems may have been responsible for the loss of roots. These results indicate that water lettuce shows potential for bioindication and phytoremediation of As-contaminated aquatic environments.

Moonlighting kinases with guanylate cyclase activity can tune regulatory signal networks

KAUST Repository

Irving, Helen R.; Kwezi, Lusisizwe; Wheeler, Janet I.; Gehring, Christoph A

2012-01-01

Guanylate cyclase (GC) catalyzes the formation of cGMP and it is only recently that such enzymes have been characterized in plants. One family of plant GCs contains the GC catalytic center encapsulated within the intracellular kinase domain of leucine rich repeat receptor like kinases such as the phytosulfokine and brassinosteroid receptors. In vitro studies show that both the kinase and GC domain have catalytic activity indicating that these kinase-GCs are examples of moonlighting proteins with dual catalytic function. The natural ligands for both receptors increase intracellular cGMP levels in isolated mesophyll protoplast assays suggesting that the GC activity is functionally relevant. cGMP production may have an autoregulatory role on receptor kinase activity and/or contribute to downstream cell expansion responses. We postulate that the receptors are members of a novel class of receptor kinases that contain functional moonlighting GC domains essential for complex signaling roles.

Ozone-induced changes in the chloroplast structure of conifer needles, and their use in ozone diagnostics

International Nuclear Information System (INIS)

Kivimaeenpaeae, M.; Sellden, G.; Sutinen, S.

2005-01-01

Ozone induces characteristic symptoms in the chloroplasts of the needles of several coniferous species. Chloroplasts are (1) reduced in size and (2) the stroma is electron dense. Moreover (3) these chloroplast alterations are more pronounced in the outer mesophyll cell layers and in the upper side of the needle compared to the inner layers and lower side. The syndrome, including the three symptoms (1)-(3), is found in the green needles of Scots pine and Norway spruce not only in the experimental fumigations, but also in mature trees in the field, and has potential for diagnosis of ozone stress. For sound ozone diagnostics all three symptoms must be present in the samples studied. The symptoms in relation to needle anatomy and physiology is discussed, and recommendations for sampling and analysis are given. - Ozone-induced alterations in chloroplast structure of conifer needles are reviewed, and recommendations for field monitoring given

Moonlighting kinases with guanylate cyclase activity can tune regulatory signal networks

KAUST Repository

Irving, Helen R.

2012-02-01

Guanylate cyclase (GC) catalyzes the formation of cGMP and it is only recently that such enzymes have been characterized in plants. One family of plant GCs contains the GC catalytic center encapsulated within the intracellular kinase domain of leucine rich repeat receptor like kinases such as the phytosulfokine and brassinosteroid receptors. In vitro studies show that both the kinase and GC domain have catalytic activity indicating that these kinase-GCs are examples of moonlighting proteins with dual catalytic function. The natural ligands for both receptors increase intracellular cGMP levels in isolated mesophyll protoplast assays suggesting that the GC activity is functionally relevant. cGMP production may have an autoregulatory role on receptor kinase activity and/or contribute to downstream cell expansion responses. We postulate that the receptors are members of a novel class of receptor kinases that contain functional moonlighting GC domains essential for complex signaling roles.

Uptake, sequestration and tolerance of cadmium at cellular levels in the hyperaccumulator plant species Sedum alfredii

Energy Technology Data Exchange (ETDEWEB)

Tian, Shengke; Xie, Ruohan; Wang, Haixin; Hu, Yan; Hou, Dandi; Liao, Xingcheng; Brown, Patrick H.; Yang, Hongxia; Lin, Xianyong; Labavitch, John M.; Lu, Lingli

2017-04-01

Sedum alfredii is one of a few plant species known to hyperaccumulate cadmium (Cd). Uptake, localization, and tolerance of Cd at cellular levels in shoots were compared in hyperaccumulating (HE) and non-hyperaccumulating (NHE) ecotypes of Sedum alfredii. X-ray fluorescence images of Cd in stems and leaves showed only a slight Cd signal restricted within vascular bundles in the NHEs, while enhanced localization of Cd, with significant tissue- and age-dependent variations, was detected in HEs. In contrast to the vascular-enriched Cd in young stems, parenchyma cells in leaf mesophyll, stem pith and cortex tissues served as terminal storage sites for Cd sequestration in HEs. Kinetics of Cd transport into individual leaf protoplasts of the two ecotypes showed little difference in Cd accumulation. However, far more efficient storage of Cd in vacuoles was apparent in HEs. Subsequent analysis of cell viability and hydrogen peroxide levels suggested that HE protoplasts exhibited higher resistance to Cd than those of NHE protoplasts. These results suggest that efficient sequestration into vacuoles, as opposed to rapid transport into parenchyma cells, is a pivotal process in Cd accumulation and homeostasis in shoots of HE S. alfredii. This is in addition to its efficient root-to-shoot translocation of Cd.

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

Science.gov (United States)

Muhaidat, Riyadh; Sage, Tammy L; Frohlich, Michael W; Dengler, Nancy G; Sage, Rowan F

2011-10-01

Photosynthetic pathway characteristics were studied in nine species of Heliotropium (sensu lato, including Euploca), using assessments of leaf anatomy and ultrastructure, activities of PEP carboxylase and C₄ acid decarboxylases, and immunolocalization of ribulose 1·5-bisphosphate carboxylase/oxygenase (Rubisco) and the P-subunit of glycine decarboxylase (GDC). Heliotropium europaeum, Heliotropium calcicola and Heliotropium tenellum are C₃ plants, while Heliotropium texanum and Heliotropium polyphyllum are C₄ species. Heliotropium procumbens and Heliotropium karwinskyi are functionally C₃, but exhibit 'proto-Kranz' anatomy where bundle sheath (BS) cells are enlarged and mitochondria primarily occur along the centripetal (inner) wall of the BS cells; GDC is present throughout the leaf. Heliotropium convolvulaceum and Heliotropium greggii are C₃--C₄ intermediates, with Kranz-like enlargement of the BS cells, localization of mitochondria along the inner BS wall and a loss of GDC in the mesophyll (M) tissue. These C₃--C₄ species of Heliotropium probably shuttle photorespiratory glycine from the M to the BS tissue for decarboxylation. Heliotropium represents an important new model for studying C₄ evolution. Where existing models such as Flaveria emphasize diversification of C₃--C₄ intermediates, Heliotropium has numerous C₃ species expressing proto-Kranz traits that could represent a critical initial phase in the evolutionary origin of C₄ photosynthesis. © 2011 Blackwell Publishing Ltd.

Carbon dioxide and water transport through plant aquaporins.

Science.gov (United States)

Groszmann, Michael; Osborn, Hannah L; Evans, John R

2017-06-01

Aquaporins are channel proteins that function to increase the permeability of biological membranes. In plants, aquaporins are encoded by multigene families that have undergone substantial diversification in land plants. The plasma membrane intrinsic proteins (PIPs) subfamily of aquaporins is of particular interest given their potential to improve plant water relations and photosynthesis. Flowering plants have between 7 and 28 PIP genes. Their expression varies with tissue and cell type, through development and in response to a variety of factors, contributing to the dynamic and tissue specific control of permeability. There are a growing number of PIPs shown to act as water channels, but those altering membrane permeability to CO 2 are more limited. The structural basis for selective substrate specificities has not yet been resolved, although a few key amino acid positions have been identified. Several regions important for dimerization, gating and trafficking are also known. PIP aquaporins assemble as tetramers and their properties depend on the monomeric composition. PIPs control water flux into and out of veins and stomatal guard cells and also increase membrane permeability to CO 2 in mesophyll and stomatal guard cells. The latter increases the effectiveness of Rubisco and can potentially influence transpiration efficiency. © 2016 John Wiley & Sons Ltd.

EFFECTS OF NUTRIENT PRE-EXPOSURE ON ATRAZINE TOXICITY TO VALLISNERIA AMERICANA MICHX. (WILD CELERY)

Science.gov (United States)

The importance of aquatic plants as a valued component of coastal environments is receiving increased attention in the public policy and scientific community. Unfortunately, the fact remains that the effects of anthropogenic contaminants on both emergent (marsh) and submerged aq...

Improved efficiency of plant regeneration from protoplasts of eggplant Solanum melongena L.

Science.gov (United States)

Guri, A; Izhar, S

1984-12-01

Eggplant (Solanum melongena L.) mesophyll protoplasts were obtained from in vitro growing plants of line 410 and cv. 'Classic'. Relatively high (15%) plating efficiency was achieved using petri dishes with alternate quadrants containing reservoir medium (R medium + 1% activated charcoal) and culture medium. Shoot regeneration occurred within 6 weeks following initiation of protoplast culture.

The different fates of mitochondria and chloroplasts during dark-induced senescence in Arabidopsis leaves.

Science.gov (United States)

Keech, Olivier; Pesquet, Edouard; Ahad, Abdul; Askne, Anna; Nordvall, Dag; Vodnala, Sharvani Munender; Tuominen, Hannele; Hurry, Vaughan; Dizengremel, Pierre; Gardeström, Per

2007-12-01

Senescence is an active process allowing the reallocation of valuable nutrients from the senescing organ towards storage and/or growing tissues. Using Arabidopsis thaliana leaves from both whole darkened plants (DPs) and individually darkened leaves (IDLs), we investigated the fate of mitochondria and chloroplasts during dark-induced leaf senescence. Combining in vivo visualization of fates of the two organelles by three-dimensional reconstructions of abaxial parts of leaves with functional measurements of photosynthesis and respiration, we showed that the two experimental systems displayed major differences during 6 d of dark treatment. In whole DPs, organelles were largely retained in both epidermal and mesophyll cells. However, while the photosynthetic capacity was maintained, the capacity of mitochondrial respiration decreased. In contrast, IDLs showed a rapid decline in photosynthetic capacity while maintaining a high capacity for mitochondrial respiration throughout the treatment. In addition, we noticed an unequal degradation of organelles in the different cell types of the senescing leaf. From these data, we suggest that metabolism in leaves of the whole DPs enters a 'stand-by mode' to preserve the photosynthetic machinery for as long as possible. However, in IDLs, mitochondria actively provide energy and carbon skeletons for the degradation of cell constituents, facilitating the retrieval of nutrients. Finally, the heterogeneity of the degradation processes involved during senescence is discussed with regard to the fate of mitochondria and chloroplasts in the different cell types.

Stereology, an unbiased methodological approach to study plant anatomy and cytology: Past, present and future

Czech Academy of Sciences Publication Activity Database

Kubínová, Lucie; Radochová, Barbora; Lhotáková, Z.; Kubínová, Z.; Albrechtová, J.

2017-01-01

Roč. 36, č. 3 (2017), s. 187-205 ISSN 1580-3139 R&D Projects: GA MŠk(CZ) LM2015062 Institutional support: RVO:67985823 Keywords : chloroplast * confocal microscopy * leaf anatomy * mesophyll * stereological methods * systematic uniform random sampling Subject RIV: FS - Medical Facilities ; Equipment OBOR OECD: Medical laboratory technology (including laboratory samples analysis Impact factor: 1.135, year: 2016

Differential methods of localisation of fungal endophytes in the seagrasses

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S. Raja

2016-07-01

Full Text Available Sections of three seagrass species (Halophila ovalis, Cymodocea serrulata and Halodule pinifolia were assessed for endophytes based on differential staining using light and fluorescence microscopy method. Acridine orange and aniline blue detected endophytic fungi in 20% and 10% of the segments, respectively, whereas lactophenol cotton blue was more sensitive to detect the fungal hyphae in 70% of the segments. Hyphae were the principal fungal structures generally observed under the cuticle, within the epidermal cells, mesophyll (Parenchyma cells and occasionally within the vascular tissue that varied in type, size and location within the leaf tissue. Present study also recorded the sporulation for the first time from the seagrass endophytes. Successfully amplified products of the ITS region of endophytic fungal DNA, directly from seagrass tissue and also from culture-dependent fungal DNA clearly depicted the presence of endophytic fungi in H. ovalis with two banding patterns (903 and 1381 bp confirming the presence of two dominant fungal genera. The fingerprinting of endophytic fungal community within the seagrass tissue was assessed using denaturing gradient gel electrophoresis (DGGE that derived with multiple bands that clarified the presence of more than one taxon within the seagrass tissue.

LEAF ANATOMICAL VARIATION IN RELATION TO STRESS TOLERANCE AMONG SOME WOODY SPECIES ON THE ACCRA PLAINS OF GHANA

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DZOMEKU BELOVED MENSAH

2012-12-01

Full Text Available Leaf anatomical study was conducted on some woody species on the Accra Plains of Ghana. Leaf epidermal strips and transverse sections were mounted in Canada balsam and studied. The anatomical studies revealed numerous stomata on the lower epidermis of Azadirachta indica. The anatomical studies revealed the presence of thick cuticles, double-layered palisade mesophyll in most species and the presence of epidermal hairs in some species. Ficus capensis showed the presence of cystolith in the lower epidermis whereas Zanthoxylum zanthoxyloides showed the presence of mucilage gland in the upper epidermis. Epidermal cell of Chromolaena odorata are very large with undulating cell walls. The species studied had various adaptive anatomical features. The stomatal frequency of Azadirachta indica was very high. With the exception of Chromolaena odorata the stomatal frequencies of the species were relatively high. The stomatal dimensions showed that most of the species maintained constant stomatal length during the study period except Griffonia simplicifolia that increased the stomatal width during the afternoon. Unlike Morinda lucida, Griffonia simplicifolia and Chromolaena odorata, that showed reduction in the breadth of stomata, the other species maintained constant stomatal width.

Mesophyll cell-sucking herbivores (Cicadellidae: Typhlocybinae) on rainforest trees in New Guinea: local and regional diversity of a taxonomically unexplored guild

Czech Academy of Sciences Publication Activity Database

Baje, L.; Stewart, A. J. A.; Novotný, Vojtěch

2014-01-01

Roč. 39, č. 3 (2014), s. 325-333 ISSN 0307-6946 R&D Projects: GA ČR GAP505/10/0673; GA ČR GA206/09/0115; GA MŠk(CZ) LH11008 Grant - others:National Science Foundation(US) DEB 0515678; European Social Fund(CZ) CZ.1.07/2.3.00/20.0064; UK Darwin Initiative(GB) 14/054 Institutional support: RVO:60077344 Keywords : Auchenorrhyncha * effective specialisation * food webs Subject RIV: EH - Ecology, Behaviour Impact factor: 1.699, year: 2014 http://onlinelibrary.wiley.com/doi/10.1111/een.12104/pdf

Effects of ozone on the foliar histology of the mastic plant (Pistacia lentiscus L.)

Energy Technology Data Exchange (ETDEWEB)

Reig-Arminana, J.; Calatayud, V.; Cervero, J.; Garcia-Breijo, F.J.; Ibars, A.; Sanz, M.J

2004-11-01

An open-top chamber study was conducted to investigate the tissue and cellular-level foliar effects of ozone (O{sub 3}) on a Mediterranean evergreen species, the mastic plant (Pistacia lentiscus L.). Plants were exposed at three different O{sub 3} levels, and leaf samples were collected periodically from the beginning of the exposure. Although no visible foliar injury was evident, alterations of the plastids and vacuoles in the mesophyll were observed. Senescence processes were accelerated with an anomalous stacking of tannin vacuoles, and a reduction in the size and number of the chloroplasts. Overall, most of the modifications induced by O{sub 3} were consistent with previously reported observations on deciduous broadleaf species, with the exception of alterations in the cells covering the secretory channels, reported here as a new finding. Comments on the feasibility of using microscopy to validate O{sub 3} related field observations and subtle foliar injury are also given.

A natural variant of NAL1, selected in high-yield rice breeding programs, pleiotropically increases photosynthesis rate

Science.gov (United States)

Takai, Toshiyuki; Adachi, Shunsuke; Taguchi-Shiobara, Fumio; Sanoh-Arai, Yumiko; Iwasawa, Norio; Yoshinaga, Satoshi; Hirose, Sakiko; Taniguchi, Yojiro; Yamanouchi, Utako; Wu, Jianzhong; Matsumoto, Takashi; Sugimoto, Kazuhiko; Kondo, Katsuhiko; Ikka, Takashi; Ando, Tsuyu; Kono, Izumi; Ito, Sachie; Shomura, Ayahiko; Ookawa, Taiichiro; Hirasawa, Tadashi; Yano, Masahiro; Kondo, Motohiko; Yamamoto, Toshio

2013-01-01

Improvement of leaf photosynthesis is an important strategy for greater crop productivity. Here we show that the quantitative trait locus GPS (GREEN FOR PHOTOSYNTHESIS) in rice (Oryza sativa L.) controls photosynthesis rate by regulating carboxylation efficiency. Map-based cloning revealed that GPS is identical to NAL1 (NARROW LEAF1), a gene previously reported to control lateral leaf growth. The high-photosynthesis allele of GPS was found to be a partial loss-of-function allele of NAL1. This allele increased mesophyll cell number between vascular bundles, which led to thickened leaves, and it pleiotropically enhanced photosynthesis rate without the detrimental side effects observed in previously identified nal1 mutants, such as dwarf plant stature. Furthermore, pedigree analysis suggested that rice breeders have repeatedly selected the high-photosynthesis allele in high-yield breeding programs. The identification and utilization of NAL1 (GPS) can enhance future high-yield breeding and provides a new strategy for increasing rice productivity. PMID:23985993

Anatomy of the vegetative organs at Syngonium podophyllum Schott.

Directory of Open Access Journals (Sweden)

Monica SIPOS

2009-11-01

Full Text Available This article has as an objective establishing the structure of the vegetative organs at Syngonium podophyllum. The structure is specific for herbaceous monocotyledonous: root has a primary structure, the stem primary structure is an intermediary form between an aerial stem and a rhizome (the presence in a fundamental parenchyma of the colaterally closed vascular bundles and the leptocentric ones. The leaf petiole has the suberified epidermis. It is characterised by the colaterally closed vascular bundles disposed peripherically and extremely well protected by the sclerenchyma and in the centre of the petiole, in the fundamental parenchyma, the same type of fascicles are placed. The middrib has a structure similar to that of the petiole. The leaf mesophyll is homogenous. The parenchyma of aerial vegetative organs of this species is crossed by aeripherous channels and their cells contain calcium oxalate crystals. The leaf is amphistomatic, the stomatic complexes are an amarilidaceous type, tetracitic or hexacitic.

The variability of leaf anatomical characteristics of Solanum nigrum L. (Solana-les, Solanaceae from different habitats

Directory of Open Access Journals (Sweden)

Krstić Lana N.

2002-01-01

Full Text Available In Europe on the whole as well as in Yugoslavia, the most widespread weed species from the genus Solanum is Solanum nigrum L. Since this species inhabits different habitats, it developed several ways of adaptation to environmental conditions. The influence of ecological factors on plant organism and resulting plant adaptations are most evident in leaf morphology and anatomy. Therefore, the anatomical structure of leaves and leaf epidermal tissue of S. nigrum was analyzed and compared among plants that originated from different habitats, in order to determine leaf structural adaptations. S. nigrum lamina has the mesomorphic structure with some xero-heliomorphic adaptations. The differences in stomata number, number of hairs, thickness of lamina, palisade and spongy tissue, as well as the size of mesophyll cells have been noticed. The highest values for most of the parameters have been recorded for the plants from cultivated soil. Largest variations of the examined characters were found for the leaves from ruderal habitats, where environmental conditions are most variable.

Final Environmental Assessment for the California Space Center at Vandenberg Air Force Base, California

Science.gov (United States)

2010-06-02

rooted , mesophylic plant species that Chapter 3. Affected Environment Final Environmental Assessment - California Space Center, Vandenberg Air...Chapter 3. Affected Environment 3-12 Final Environmental Assessment - California Space Center, Vandenberg Air Force Base the root and debris zone of the...protruding objects, slippery soils or mud, and biological hazards including vegetation (i.e. poison oak and stinging nettle ), animals (i.e. insects

Programmatic Environmental Assessment, 2007 General Plan for the Main Cantonment and the South Base Cantonment at Vandenberg Air Force Base, California

Science.gov (United States)

2008-05-05

cunicularia) and mountain plover (Charadrius montanus). Central Coast Scrub This vegetation type is characterized by shallow- rooted , mesophylic plant...Wild blackberry (Rubus ursinus), mugwort (Artemisia douglasiana), and stinging nettle (Urtica dioica) are common understory components of...feed until maturation. Upon maturation, larvae burrow into the soil and pupate, usually within the root and debris zone of the host plant (Mattoni

Environmental Assessment for the California Space Center at Vandenberg Air Force Base, California

Science.gov (United States)

2010-04-08

shallow- rooted , mesophylic plant species that Chapter 3. Affected Environment Final Draft Environmental Assessment - California Space Center...buckwheat flowers and buds where the larvae feed until maturation. Upon maturation larvae burrow into the soil and pupate, usually within the root and...terrain, sharp or protruding objects, slippery soils or mud, and biological hazards including vegetation (i.e. poison oak and stinging nettle

Climate change and fire regimes in the Sierra de Manantlan, Mexico

Science.gov (United States)

Brooke A. Cassell; Ernesto Alvarado; Emily Heyerdahl; Diego Perez-Salicrup; Enrique Jardel-Pelaez

2010-01-01

Fire has been attributed as one of the most influential factors in vegetation community and succession in the Sierra de Manantlán Biosphere Reserve in Jalisco and Colima, México. A mosaic of low, mixed and high severity fire regimes characterizes the landscape with ecosystems ranging from mesophyllous mountain forest to higher elevation pine and oak forest. Research...

In vitro growth and leaf anatomy of Cattleya walkeriana (Gardner, 1839 grown in natural ventilation system

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Adriano Bortolotti da Silva

2014-12-01

Full Text Available Natural ventilation system facilitates gaseous exchanges in in vitro plants promoting changes in the leaf tissue, which can be evaluated through the leaf anatomy, and it allows a cultivation closer to the photoautrophic micropropagation. The objective of this work was to evaluate the effects on in vitro growth and on the leaf anatomy of Cattleya walkeriana grown in natural and conventional ventilation system with different concentrations of sucrose (0; 15; 30 and 45 L-1 combined with different cultivation systems (conventional micropropagation and natural ventilation system. The culture medium was composed of MS salts, solidified with 7 g L-1 of agar and pH adjusted to 5.8. Forty milliliters of culture medium were distributed in 250 mL flasks, autoclaved at 120 ºC for 20 minutes. The greater plant growth, as well as the greater thickness of the mesophyll was observed with the use of 20 g L-1 sucrose in natural ventilation system. Plants grown in natural ventilation system showed a thicker leaf mesophyll, which is directly related to photoautotrophic crops. The natural ventilation system induced more elliptical stomata and probably more functional formats.

Nitroaromatic detection and infrared communication from wild-type plants using plant nanobionics

Science.gov (United States)

Wong, Min Hao; Giraldo, Juan P.; Kwak, Seon-Yeong; Koman, Volodymyr B.; Sinclair, Rosalie; Lew, Tedrick Thomas Salim; Bisker, Gili; Liu, Pingwei; Strano, Michael S.

2017-02-01

Plant nanobionics aims to embed non-native functions to plants by interfacing them with specifically designed nanoparticles. Here, we demonstrate that living spinach plants (Spinacia oleracea) can be engineered to serve as self-powered pre-concentrators and autosamplers of analytes in ambient groundwater and as infrared communication platforms that can send information to a smartphone. The plants employ a pair of near-infrared fluorescent nanosensors--single-walled carbon nanotubes (SWCNTs) conjugated to the peptide Bombolitin II to recognize nitroaromatics via infrared fluorescent emission, and polyvinyl-alcohol functionalized SWCNTs that act as an invariant reference signal--embedded within the plant leaf mesophyll. As contaminant nitroaromatics are transported up the roots and stem into leaf tissues, they accumulate in the mesophyll, resulting in relative changes in emission intensity. The real-time monitoring of embedded SWCNT sensors also allows residence times in the roots, stems and leaves to be estimated, calculated to be 8.3 min (combined residence times of root and stem) and 1.9 min mm-1 leaf, respectively. These results demonstrate the ability of living, wild-type plants to function as chemical monitors of groundwater and communication devices to external electronics at standoff distances.

Nitroaromatic detection and infrared communication from wild-type plants using plant nanobionics.

Science.gov (United States)

Wong, Min Hao; Giraldo, Juan P; Kwak, Seon-Yeong; Koman, Volodymyr B; Sinclair, Rosalie; Lew, Tedrick Thomas Salim; Bisker, Gili; Liu, Pingwei; Strano, Michael S

2017-02-01

Plant nanobionics aims to embed non-native functions to plants by interfacing them with specifically designed nanoparticles. Here, we demonstrate that living spinach plants (Spinacia oleracea) can be engineered to serve as self-powered pre-concentrators and autosamplers of analytes in ambient groundwater and as infrared communication platforms that can send information to a smartphone. The plants employ a pair of near-infrared fluorescent nanosensors-single-walled carbon nanotubes (SWCNTs) conjugated to the peptide Bombolitin II to recognize nitroaromatics via infrared fluorescent emission, and polyvinyl-alcohol functionalized SWCNTs that act as an invariant reference signal-embedded within the plant leaf mesophyll. As contaminant nitroaromatics are transported up the roots and stem into leaf tissues, they accumulate in the mesophyll, resulting in relative changes in emission intensity. The real-time monitoring of embedded SWCNT sensors also allows residence times in the roots, stems and leaves to be estimated, calculated to be 8.3 min (combined residence times of root and stem) and 1.9 min mm -1 leaf, respectively. These results demonstrate the ability of living, wild-type plants to function as chemical monitors of groundwater and communication devices to external electronics at standoff distances.

Plant-herbivore interaction: dissection of the cellular pattern of Tetranychus urticae feeding on the host plant

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Nicolas Bensoussan

2016-07-01

Full Text Available The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae, is one of the most polyphagous herbivores feeding on cell contents of over 1,100 plant species including more than 150 crops. It is being established as a model for chelicerate herbivores with tools that enable tracking of reciprocal responses in plant-spider mite interactions. However, despite their important pest status and a growing understanding of the molecular basis of interactions with plant hosts, knowledge of the way mites interface with the plant while feeding and the plant damage directly inflicted by mites is lacking. Here, utilizing histology and microscopy methods, we uncovered several key features of T. urticae feeding. By following the stylet path within the plant tissue, we determined that the stylet penetrates the leaf either in between epidermal pavement cells or through a stomatal opening, without damaging the epidermal cellular layer. Our recordings of mite feeding established that duration of the feeding event ranges from several minutes to more than half an hour, during which time mites consume a single mesophyll cell in a pattern that is common to both bean and Arabidopsis plant hosts. In addition, this study determined that leaf chlorotic spots, a common symptom of mite herbivory, do not form as an immediate consequence of mite feeding. Our results establish a cellular context for the plant-spider mite interaction that will support our understanding of the molecular mechanisms and cell signaling associated with spider mite feeding.

Leaf anatomical and photosynthetic acclimation to cool temperature and high light in two winter versus two summer annuals.

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Cohu, Christopher M; Muller, Onno; Adams, William W; Demmig-Adams, Barbara

2014-09-01

Acclimation of foliar features to cool temperature and high light was characterized in winter (Spinacia oleracea L. cv. Giant Nobel; Arabidopsis thaliana (L.) Heynhold Col-0 and ecotypes from Sweden and Italy) versus summer (Helianthus annuus L. cv. Soraya; Cucurbita pepo L. cv. Italian Zucchini Romanesco) annuals. Significant relationships existed among leaf dry mass per area, photosynthesis, leaf thickness and palisade mesophyll thickness. While the acclimatory response of the summer annuals to cool temperature and/or high light levels was limited, the winter annuals increased the number of palisade cell layers, ranging from two layers under moderate light and warm temperature to between four and five layers under cool temperature and high light. A significant relationship was also found between palisade tissue thickness and either cross-sectional area or number of phloem cells (each normalized by vein density) in minor veins among all four species and growth regimes. The two winter annuals, but not the summer annuals, thus exhibited acclimatory adjustments of minor vein phloem to cool temperature and/or high light, with more numerous and larger phloem cells and a higher maximal photosynthesis rate. The upregulation of photosynthesis in winter annuals in response to low growth temperature may thus depend on not only (1) a greater volume of photosynthesizing palisade tissue but also (2) leaf veins containing additional phloem cells and presumably capable of exporting a greater volume of sugars from the leaves to the rest of the plant. © 2014 Scandinavian Plant Physiology Society.

Using Seeds to Propagate and Restore Vallisneria americana Michaux (Wild Celery) in the Chesapeake Bay

Science.gov (United States)

2007-12-01

the capacity of the plants to elongate so that the leaves can reach closer to the water surface to gather adequate light for photosynthesis . When...transplant eelgrass (Zostera marina L.) in Chesapeake Bay and the Virginia Coastal Bays, In Proc. Conf. Seagrass Restoration: Success, Failure, and

Radial oxygen loss from roots of Vallisneria spiralis L.: biogeochemical implications in eutrophic aquatic ecosystems

OpenAIRE

Soana, Elisa

2013-01-01

Eutrophication and the accumulation of organic matter have been addressed as the major factors determining the decline of benthic vegetation in impacted water bodies and the consequent loss of key ecosystemic functions. In freshwater environments the literature reports submersed macrophyte die-back events and the switch to free-floating and floating-leaved plants dominated states. Species-specific differences in macrophyte response along organic gradients are evident. Some species have develo...

Application of HB17, an Arabidopsis class II homeodomain-leucine zipper transcription factor, to regulate chloroplast number and photosynthetic capacity.

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Hymus, Graham J; Cai, Suqin; Kohl, Elizabeth A; Holtan, Hans E; Marion, Colleen M; Tiwari, Shiv; Maszle, Don R; Lundgren, Marjorie R; Hong, Melissa C; Channa, Namitha; Loida, Paul; Thompson, Rebecca; Taylor, J Philip; Rice, Elena; Repetti, Peter P; Ratcliffe, Oliver J; Reuber, T Lynne; Creelman, Robert A

2013-11-01

Transcription factors are proposed as suitable targets for the control of traits such as yield or food quality in plants. This study reports the results of a functional genomics research effort that identified ATHB17, a transcription factor from the homeodomain-leucine zipper class II family, as a novel target for the enhancement of photosynthetic capacity. It was shown that ATHB17 is expressed natively in the root quiescent centre (QC) from Arabidopsis embryos and seedlings. Analysis of the functional composition of genes differentially expressed in the QC from a knockout mutant (athb17-1) compared with its wild-type sibling revealed the over-representation of genes involved in auxin stimulus, embryo development, axis polarity specification, and plastid-related processes. While no other phenotypes were observed in athb17-1 plants, overexpression of ATHB17 produced a number of phenotypes in Arabidopsis including enhanced chlorophyll content. Image analysis of isolated mesophyll cells of 35S::ATHB17 lines revealed an increase in the number of chloroplasts per unit cell size, which is probably due to an increase in the number of proplastids per meristematic cell. Leaf physiological measurements provided evidence of improved photosynthetic capacity in 35S::ATHB17 lines on a per unit leaf area basis. Estimates of the capacity for ribulose-1,5-bisphosphate-saturated and -limited photosynthesis were significantly higher in 35S::ATHB17 lines.

Chromoplast biogenesis in Chelidonium majus petals

OpenAIRE

Nikola Ljubešić; Mercedes Wrischer

2011-01-01

The differentiation of chromoplasts, with special emphasis on the formation and the organisation of chromoplast fibrils, was followed in the petals of the greater celandine, Chelidonium majus L. Electron microscopic observations showed that, in the epidermis, differentiation of chromoplasts started from leucoplasts, while mesophyll chromoplasts originated from chloroplasts. During petal maturation, fibrils accumulated in the plastids, often arranging in a parallel fashion to form compact bire...

Quantitative imaging of radial oxygen loss from Valisneria spiralis roots with a fluorescent planar optode

Energy Technology Data Exchange (ETDEWEB)

Han, Chao [State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008 (China); Ren, Jinghua [Geological Survey of Jiangsu Province, Nanjing 210018 (China); Tang, Hao [State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008 (China); Xu, Di, E-mail: dxu@niglas.ac.cn [State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008 (China); Xie, Xianchuan, E-mail: xchxie@nju.edu.cn [State Key Laboratory of Pollution Control and Resource Reuse, Center for Hydroscience Research, School of the Environment, Nanjing University, Nanjing 210093 (China)

2016-11-01

Oxygen (O{sub 2}) availability within the sediment–root interface is critical to the survival of macrophytes in O{sub 2}-deficient sediment; however, our knowledge of the fine-scale impact of macrophyte roots upon the spatiotemporal dynamics of O{sub 2} is relatively limited. In this study, a non-invasive imaging technology was utilized to map O{sub 2} micro-distribution around Vallisneria spiralis. Long-term imaging results gathered during a 36 day-period revealed an abundance of O{sub 2} spatiotemporal patterns ranging from 0 to 250 μmol L{sup −} {sup 1}. The root-induced O{sub 2} leakage and consequent oxygenated area were stronger in the vicinity of the basal root compared to that found in the root tip. The O{sub 2} images revealed V. spiralis exhibited radial O{sub 2} loss (ROL) along the entire root, and the O{sub 2} distribution along the root length showed a high degree of small-scale spatial heterogeneity decreasing from 80% at the basal root surface to 10% at the root tip. The oxygenated zone area around the roots increased as O{sub 2} levels increased with root growth and irradiance intensities ranging from 0 to 216 μmol photons m{sup −} {sup 2} s{sup −} {sup 1}. A weak ROL measuring < 20% air saturation around the basal root surface was maintained in darkness, which was presumably attributed to the O{sub 2} supply from overlying water via plant aerenchyma. The estimated total O{sub 2} release to the rhizosphere of V. spiralis was determined to range from 8.80 ± 7.32 to 30.34 ± 17.71 nmol m{sup −} {sup 2} s{sup −} {sup 1}, which is much higher than many other macrophyte species. This O{sub 2} release may be an important contribution to the high-capacity of V. spiralis for quickly colonizing anaerobic sediment. - Highlights: • Planar imaging method was used to map O2 micro-distribution. • Highly dynamic rhizospheric O2-spatiotemporal distribution was observed. • O{sub 2} leakage along the entire root of Vallisneria spirals were

Anatomical, biochemical, and photosynthetic responses to recent allopolyploidy in Glycine dolichocarpa (Fabaceae).

Science.gov (United States)

Coate, Jeremy E; Luciano, Amelia K; Seralathan, Vasu; Minchew, Kevin J; Owens, Tom G; Doyle, Jeff J

2012-01-01

Previous studies have shown that polyploidy has pronounced effects on photosynthesis. Most of these studies have focused on synthetic or recently formed autopolyploids, and comparatively little is known about the integrated effects of natural allopolyploidy, which involves hybridity and genome doubling and often incorporates multiple genotypes through recurrent origins and lineage recombination. Glycine dolichocarpa (designated T2) is a natural allotetraploid with multiple origins. We quantified 21 anatomical, biochemical, and physiological phenotypes relating to photosynthesis in T2 and its diploid progenitors, G. tomentella (D3) and G. syndetika (D4). To assess how direction of cross affects these phenotypes, we included three T2 accessions having D3-like plastids (T2(D3)) and two accessions having D4-like plastids (T2(D4)). T2 accessions were transgressive (more extreme than any diploid accession) for 17 of 21 phenotypes, and species means differed significantly in T2 vs. both progenitors for four of 21 phenotypes (higher for guard cell length, electron transport capacity [J(max)] per palisade cell, and J(max) per mesophyll cell; lower for palisade cells per unit leaf area). Within T2, four of 21 parameters differed significantly between T2(D3) and T2(D4) (palisade cell volume; chloroplast number and volume per unit leaf area; and J(max) per unit leaf area). T2 is characterized by transgressive photosynthesis-related phenotypes (including an ca. 2-fold increase in J(max) per cell), as well as by significant intraspecies variation correlating with plastid type. These data indicate prominent roles for both nucleotypic effects and cytoplasmic factors in photosynthetic responses to allopolyploidy.

Coordination of Leaf Photosynthesis, Transpiration, and Structural Traits in Rice and Wild Relatives (Genus Oryza).

Science.gov (United States)

Giuliani, Rita; Koteyeva, Nuria; Voznesenskaya, Elena; Evans, Marc A; Cousins, Asaph B; Edwards, Gerald E

2013-07-01

The genus Oryza, which includes rice (Oryza sativa and Oryza glaberrima) and wild relatives, is a useful genus to study leaf properties in order to identify structural features that control CO(2) access to chloroplasts, photosynthesis, water use efficiency, and drought tolerance. Traits, 26 structural and 17 functional, associated with photosynthesis and transpiration were quantified on 24 accessions (representatives of 17 species and eight genomes). Hypotheses of associations within, and between, structure, photosynthesis, and transpiration were tested. Two main clusters of positively interrelated leaf traits were identified: in the first cluster were structural features, leaf thickness (Thick(leaf)), mesophyll (M) cell surface area exposed to intercellular air space per unit of leaf surface area (S(mes)), and M cell size; a second group included functional traits, net photosynthetic rate, transpiration rate, M conductance to CO(2) diffusion (g(m)), stomatal conductance to gas diffusion (g(s)), and the g(m)/g(s) ratio.While net photosynthetic rate was positively correlated with gm, neither was significantly linked with any individual structural traits. The results suggest that changes in gm depend on covariations of multiple leaf (S(mes)) and M cell (including cell wall thickness) structural traits. There was an inverse relationship between Thick(leaf) and transpiration rate and a significant positive association between Thick(leaf) and leaf transpiration efficiency. Interestingly, high g(m) together with high g(m)/g(s) and a low S(mes)/g(m) ratio (M resistance to CO(2) diffusion per unit of cell surface area exposed to intercellular air space) appear to be ideal for supporting leaf photosynthesis while preserving water; in addition, thick M cell walls may be beneficial for plant drought tolerance.

Remote sensing of changes in morphology and physiology of trees under stress

Science.gov (United States)

Olson, C. E., Jr.; Rohde, W. G.; Ward, J. M.

1970-01-01

Results of continuing studies of forest trees subjected to varying types of stress are reported. Both greenhouse and field studies are included. Greenhouse work with tree seedlings exposed to varying levels of NaCl and CaCl2 in the soil indicated that, in the initial stages, palisade cells shrink and the amount of air space in the leaf increases. As the severity of damage increases, the cells of the spongy mesophyll shrink and flatten, and the amount of air space in the leaf decreases. Statistical analysis of foliar reflectance and associated moisture content data led to a series of regression equations for predicting foliar moisture content from reflectance data. Equations were calculated for three species, yellow birch (Betula alleghaniensis Britton), sugar maple (Acer saccharum Marsh.) and white ash (Fraxinus americana L.) having multiple correlation coefficients of 0.98, 0.94 and 0.93 respectively. Interpretation of multispectral imagery of the Ann Arbor Forestry Test Site (NASA Site 190) provided evidence that infections of Fomes annosus can be detected in the early stages. Infections of two needle cast diseases were also detected in conifer plantations in the test site. A study of automatic interpretation of multispectral scanner imagery for tree species recognition provided encouraging results.

Physiological Responses to Cadmium, Nickel and their Interaction in the Seedlings of Two Maize (Zea mays L. Cultivars

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Pavlovkin Ján

2016-12-01

Full Text Available In the leaves of maize seedlings, cultivars Premia and Blitz, the relatively low 2 μmol/L concentration of cadmium (Cd, nickel (Ni, or both metals acting simultaneously (Cd +Ni for 72 h, induced a significant metal accumulation, decrease in total K+ content, reduction of light-induced membrane electrical potential (EM repolarisation in mesophyll cells and changes of ascorbic acid (AsA, dehydroascorbic acid (DHA and glutathione (GSH content. Shoot growth and the values of resting EM did not change significantly. Increased K+ leakage, from the leaves, and lipid peroxidation accompanied by increase of TBA-reactive substances (TBARS were found only in cv. Blitz exposed to Cd + Ni. This indicates a capability of high leaf-cell anti-oxidant pool to ameliorate the toxic effects on plasma membrane of single ions in both cultivars, and of Cd + Ni only in cv. Premia. The decreased total content of K+ in leaves in all variants indicated repressing the K+ uptake and/or distribution to the shoots. Under anoxia, the magnitude of the repolarisation obtained after switching on the light was smaller in Cd-treated cultivar Premia than in the controls, and this also occurred in Ni- and Cd + Ni-treated cultivar Blitz.

Vesicles Are Persistent Features of Different Plastids.

Science.gov (United States)

Lindquist, Emelie; Solymosi, Katalin; Aronsson, Henrik

2016-10-01

Peripheral vesicles in plastids have been observed repeatedly, primarily in proplastids and developing chloroplasts, in which they are suggested to function in thylakoid biogenesis. Previous observations of vesicles in mature chloroplasts have mainly concerned low temperature pretreated plants occasionally treated with inhibitors blocking vesicle fusion. Here, we show that such vesicle-like structures occur not only in chloroplasts and proplastids, but also in etioplasts, etio-chloroplasts, leucoplasts, chromoplasts and even transforming desiccoplasts without any specific pretreatment. Observations are made both in C3 and C4 species, in different cell types (meristematic, epidermis, mesophyll, bundle sheath and secretory cells) and different organs (roots, stems, leaves, floral parts and fruits). Until recently not much focus has been given to the idea that vesicle transport in chloroplasts could be mediated by proteins, but recent data suggest that the vesicle system of chloroplasts has similarities with the cytosolic coat protein complex II system. All current data taken together support the idea of an ongoing, active and protein-mediated vesicle transport not only in chloroplasts but also in other plastids, obviously occurring regardless of chemical modifications, temperature and plastid developmental stage. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Hpa1 harpin needs nitroxyl terminus to promote vegetative growth and leaf photosynthesis in Arabidopsis.

Science.gov (United States)

Li, Xiaojie; Han, Liping; Zhao, Yanying; You, Zhenzhen; Dong, Hansong; Zhang, Chunling

2014-03-01

Hpa1 is a harpin protein produced by Xanthomonas oryzae, an important bacterial pathogen of rice, and has the growth-promoting activity in plants. To understand the molecular basis for the function of Hpa1, we generated an inactive variant protein, Hpa1 delta NT, by deleting the nitroxyl-terminal region of the Hpa1 sequence and compared Hpa1 delta NT with the full-length protein in terms of the effects on vegetative growth and related physiological responses in Arabidopsis. When Hpa1 was applied to plants, it acted to enhance the vegetative growth but did not affect the floral development. Enhanced plant growth was accompanied by induced expression of growth-promoting genes in plant leaves. The growth-promoting activity of Hpa1 was further correlated with a physiological consequence shown as promoted leaf photosynthesis as a result of facilitated CO2 conduction through leaf stomata and mesophyll cells. On the contrary, plant growth, growth-promoting gene expression, and the physiological consequence changed little in response to the Hpa1 delta NT treatment. These analyses suggest that Hpa1 requires the nitroxyl-terminus to facilitate CO2 transport inside leaf cells and promote leaf photosynthesis and vegetative growth of the plant.

Environmental Assessment for Repairs and Replacement of Overhead Electrical Line, Feeders N1, N3, and N6 Vandenberg Air Force Base, California

Science.gov (United States)

2011-08-22

along almost the entire coast of California. Shallow- rooted , mesophyllic plant species that are often drought-deciduous and summer-dormant... root and debris zone of the host plant (Mattoni 1992). Pupae remain in diapause until at least the following flight season. The number of adult...Maschner et al. 1991; Snethkamp and Munns 1991; Lebow 2001; Nettles and Hamilton 2008. 1149/H Location/ lithic scatter/ historic ranch N1, N3 Unevaluated

Final Environmental Assessment: Falcon 9 and Falcon 9 Heavy Launch Vehicle Programs from Space Launch Complex 4 East at Vandenberg Air Force Base, California

Science.gov (United States)

2011-03-01

Central Coast Scrub Central coast scrub (CCS) is characterized by shallow- rooted , mesophylic plant species that are often drought-deciduous and summer...maturation larvae burrow into the soil and pupate, usually within the root and debris zone of the host plant (Mattoni 1992; Pratt and Ballmer, pers. obs...sharp or protruding objects, slippery soils or mud, and biological hazards including vegetation (i.e. poison oak and stinging nettle ), animals (i.e

Responses of photosynthetic parameters to drought in subtropical forest ecosystem of China

Science.gov (United States)

Zhou, Lei; Wang, Shaoqiang; Chi, Yonggang; Li, Qingkang; Huang, Kun; Yu, Quanzhou

2015-12-01

The mechanism underlying the effect of drought on the photosynthetic traits of leaves in forest ecosystems in subtropical regions is unclear. In this study, three limiting processes (stomatal, mesophyll and biochemical limitations) that control the photosynthetic capacity and three resource use efficiencies (intrinsic water use efficiency (iWUE), nitrogen use efficiency (NUE) and light use efficiency (LUE)), which were characterized as the interactions between photosynthesis and environmental resources, were estimated in two species (Schima superba and Pinus massoniana) under drought conditions. A quantitative limitation analysis demonstrated that the drought-induced limitation of photosynthesis in Schima superba was primarily due to stomatal limitation, whereas for Pinus massoniana, both stomatal and non-stomatal limitations generally exhibited similar magnitudes. Although the mesophyll limitation represented only 1% of the total limitation in Schima superba, it accounted for 24% of the total limitations for Pinus massoniana. Furthermore, a positive relationship between the LUE and NUE and a marginally negative relationship or trade-off between the NUE and iWUE were observed in the control plots. However, drought disrupted the relationships between the resource use efficiencies. Our findings may have important implications for reducing the uncertainties in model simulations and advancing the understanding of the interactions between ecosystem functions and climate change.

A study of the morphoanatomical characters of the leaves of Chamaecrista (L. Moench sect. Apoucouita (Leguminosae-Caesalpinioideae

Directory of Open Access Journals (Sweden)

Ítalo Antônio Cotta Coutinho

2016-01-01

Full Text Available ABSTRACT Little attention has been paid to species of Chamaecrista sect. Apoucouita (Leguminosae-Caesalpinioideae, especially regarding anatomical studies. When only vegetative material is available, the identification of such species may be difficult. Additionally, vegetative material of some species of C. sect. Apoucouita may be even harder to identify because they can resemble species of Inga Mill. (Leguminosae-Mimosoideae. The present study focused on recognizing morphoanatomical characters of leaves that are taxonomically useful for the species of C. sect. Apoucouita by employing standard anatomical techniques. The arrangement of the vascular system in the petiole/rachis, dorsiventral mesophyll, mucilage idioblasts in the epidermis of leaflets and hypostomatic leaves were some of the characters shared by all species studied. Length of the petiole, position and type of extrafloral nectaries, leaflet venation, presence and type of papillae on the epidermis of the leaflet blades and sclereids in the mesophyll were some of the characters useful in the distinction of taxa. The vascular arrangement of the petiole/rachis is a promising character in the distinction of species of C. sect. Apoucouita and Inga. Based on morphoanatomical data, the taxonomic revision of some species and varieties ascribed to C. sect. Apoucouita is suggested.

Leaf anatomical traits of non-arboreal savanna species along a gradient of tree encroachment

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Luiz Felipe Souza Pinheiro

2017-10-01

Full Text Available ABSTRACT In the Brazilian savanna (Cerrado of Brazil, fire suppression has transformed typical savanna formations (TS into forested savanna (FS due to the phenomenon of encroachment. Under encroachment, non-arboreal plants begin to receive less light due to greater tree density and canopy closure. Here we aim to evaluate if leaf anatomical traits of non-arboreal species differ according to the degree of tree encroachment at the Assis Ecological Station - São Paulo, Brazil. To this end, we evaluated leaf tissue thickness and specific leaf area (SLA in representative non-arboreal species occurring along a gradient of tree encroachment. Leaves of TS species showed a trend towards xeromorphism, with traits reported to facilitate survival under high luminosity, such as thick leaves, thick epidermis and mesophyll, and low SLA. In contrast, FS species exhibited mesomorphic leaves, with thin mesophyll and high SLA, which are able to capture diffuse light in denser environments. Thus, non-arboreal understory species with mesomorphic leaf traits should be favored in environments with denser vegetation in contrast to typical savanna species. The results suggest that typical non-arboreal savanna species would not survive under tree encroachment due to the low competitiveness of their leaf anatomical strategies in shady environments.

Responses of tropical legumes from the Brazilian Atlantic Rainforest to simulated acid rain.

Science.gov (United States)

Andrade, Guilherme C; Silva, Luzimar C

2017-07-01

We investigated the morphological and anatomical effects of simulated acid rain on leaves of two species native to the Brazilian Atlantic Rainforest: Paubrasilia echinata and Libidibia ferrea var. leiostachya. Saplings were subjected to acid rain in a simulation chamber during 10 days for 15 min daily, using H 2 SO 4 solution pH 3.0 and, in the control, deionized water. At the end of the experiment, fragments from young and expanding leaves were anatomically analyzed. Although L. ferrea var. leiostachya leaves are more hydrophobic, rain droplets remained in contact with them for a longer time, as in the hydrophilic P. echinata leaves, droplets coalesce and rapidly run off. Visual symptomatology consisted in interveinal and marginal necrotic dots. Microscopic damage found included epicuticular wax flaking, turgor loss and epidermal cell shape alteration, hypertrophy of parenchymatous cells, and epidermal and mesophyll cell collapse. Formation of a wound tissue was observed in P. echinata, and it isolated the necrosis to the adaxial leaf surface. Acid rain increased thickness of all leaf tissues except spongy parenchyma in young leaves of L. ferrea var. leiostachya, and such thickness was maintained throughout leaf expansion. To our knowledge, this is the first report of acidity causing increase in leaf tissue thickness. This could represent the beginning of cell hypertrophy, which was seen in visually affected leaf regions. Paubrasilia echinata was more sensitive, showing earlier symptoms, but the anatomical damage in L. ferrea var. leiostachya was more severe, probably due to the higher time of contact with acid solution in this species.

Laboratory tests for the phytoextraction of heavy metals from polluted harbor sediments using aquatic plants.

Science.gov (United States)

Mânzatu, Carmen; Nagy, Boldizsár; Ceccarini, Alessio; Iannelli, Renato; Giannarelli, Stefania; Majdik, Cornelia

2015-12-30

The aim of this study was to investigate the concentrations and pollution levels of heavy metals, organochlorine pesticides, and polycyclic aromatic hydrocarbons in marine sediments from the Leghorn Harbor (Italy) on the Mediterranean Sea. The phytoextraction capacity of three aquatic plants Salvinia natans, Vallisneria spiralis, and Cabomba aquatica was also tested in the removal of lead and copper, present in high concentration in these sediments. The average detectable concentrations of metals accumulated by the plants in the studied area were as follows: >3.328 ± 0.032 mg/kg dry weight (DW) of Pb and 2.641 ± 0.014 mg/kg DW of Cu for S. natans, >3.107 ± 0.034 g/kg DW for V. spiralis, and >2.400 ± 0.029 mg/kg DW for C. aquatica. The occurrence of pesticides was also analyzed in the sediment sample by gas chromatography coupled with mass spectrometry (GC/MS). Due to its metal and organic compound accumulation patterns, S. natans is a potential candidate in phytoextraction strategies. Copyright © 2015 Elsevier Ltd. All rights reserved.

A função e a estrutura dos plastídios das células parenquimatosas que envolvem os feixes vasculares da fôlha do milho

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M. M. Rhoades

1946-01-01

Full Text Available As células do invólucro parenquimatoso que envolve os feixes vasculares da fôlha do milho encerram plastídios especializados, de côr verde e relacionados com a elaboração do amido. Os grãos de amido são formados no plastídio, em regiões definidas e semelhantes a vacúolos. Um plastídio pode apresentar até 40 grãos de amido. O amido é depositado nestes plastídios apenas quando a velocidade da transferência do açúcar das células do mesófilo para o interior das células do invólucro do feixe vascular é maior do que a velocidade com a qual o açúcar passa das células do invólucro para os elementos vasculares. Os plastídios do invólucro do feixe vascular depositam amido em quantidades crescentes durante o dia. À noite, êsse amido é hidrolizado em carbohidratos solúveis e translocado, de modo que os plastídios ficam livres de amido pela manhã. Não encontramos nenhum amido nos plastídios do mesófilo nem de dia nem de noite. Os plastídios do invólucro do feixe vascular do sorgo são semelhantes em função aos do milho ; esta especialização dos plastídios do invólucro do feixe vascular pode ser típica da sub-família Panicoidex. Os plastídios do invólucro parenquimatoso do trigo, da aveia e do centeio, todos membros da sub-família Pooidex, são diferentes no tamanho e na função, comparados com os plastídios do milho e os do sorgo.The parenchyma sheath cells enclosing the vascular bundles of the corn leaf contain specialized plastids, green in color, concerned with the elaboration of starch. The starch grains are formed in definite vacuole-like regions in the plastid. A plastid may have as many as 40 starch grains. Starch is deposited in these plastids only when the rate of movement of sugar from the mesophyll cells into the bundle sheath cells is greater than the rate with which sugar passes from the sheath cells into the vascular elements. The bundle sheath plastids deposit starch increasingly during

Stem and leaf anatomy of Plectranthus neochilus Schltr., Lamiaceae

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Márcia do Rocio Duarte

Full Text Available Plectranthus neochilus Schltr. is an aromatic herb named " boldo" or " boldo-gambá" and employed for treating hepatic insufficiency and dyspepsia in folk medicine. This paper has investigated its stem and leaf anatomy, in order to contribute for the medicinal plant identification. The botanical material was prepared according to standard microtechniques. The stem has quadrangular transection and, in secondary growth at the level analyzed, shows uniseriate epidermis and numerous trichomes. The glandular ones are capitate and peltate. The former has short unicellular or long multicellular stalk and uni- or bicellular head. The latter presents short stalk and eight-celled ovoid head. The non-glandular trichomes are multicellular, uniseriate and coated with granular cuticle. It is observed angular collenchyma, cambia forming phloem outward and xylem inward, and perivascular fiber caps next to the phloem. The blade has uniseriate epidermis coated with striate cuticle, diacytic stomata on both surfaces, numerous trichomes similar to the stem ones, and homogeneous mesophyll. The midrib shows one or two collateral bundles and the petiole has many of them distributed as an open arc.

Chemical characterization of Citrus sinensis grafted on C. limonia and the effect of some isolated compounds on the growth of Xylella fastidiosa.

Science.gov (United States)

Ribeiro, Alan Bezerra; Abdelnur, Patrícia Verardi; Garcia, Cleverson Fernando; Belini, Adriana; Severino, Vanessa G Pasqualotto; da Silva, M Fátima das G F; Fernandes, João B; Vieira, Paulo C; de Carvalho, Sérgio A; de Souza, Alessandra A; Machado, Marcos A

2008-09-10

Citrus sinensis grafted on C. limonia produces a considerable number of compounds that are not common in both plants developed from germination of seeds. The chemical profile of scion and rootstock differ notably for absence in the form of flavonoids and coumarins containing C5 prenyl groups attached to the carbon atoms of aromatic and heterocyclic systems or to oxygen. Only linear pyranocoumarins xanthyletin and xanthoxyletin were found in scion. This observation indicates that the prenylated compounds once biosynthesized in the roots could have been translocated to other organs. Xylella fastidiosa colonizes the xylem of plants causing diseases on several economically important crops such as citrus variegated chlorosis (CVC). A number of flavonoids, coumarins, alkaloids, dihydrocinnamic acid derivative, anacardic acid, triterpenes, and limonoids were tested for in vitro activity on the growth of Xylella fastidiosa. Azadirachtin A was the most active. Hesperidin, which occurs in great amounts in cells of the mesophyll of the affected leaves with CVC, showed a moderate activity suggesting that it can act as a good barrier for small-size colonies from X. fastidiosa.

Optimum design for effective water transport through a double-layered porous hydrogel inspired by plant leaves

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Kim, Hyejeong; Kim, Hyeonjeong; Huh, Hyungkyu; Hwang, Hyung Ju; Lee, Sang Joon

2014-11-01

Plant leaves are generally known to have optimized morphological structure in response to environmental changes for efficient water usage. However, the advantageous features of plant leaves are not fully utilized in engineering fields yet, since the optimum design in internal structure of plant leaves is unclear. In this study, the tissue organization of the hydraulic pathways inside plant leaves was investigated. Water transport through double-layered porous hydrogel models analogous to mesophyll cells was experimentally observed. In addition, computational experiment and theoretical analysis were applied to the model systems to find the optimal design for efficient water transport. As a result, the models with lower porosity or with pores distributed widely in the structure exhibit efficient mass transport. Our theoretical prediction supports that structural features of plant leaves guarantee sufficient water supply as survival strategy. This study may provide a new framework for investigating the biophysical principles governing the morphological optimization of plant leaves and for designing microfluidic devices to enhance mass transport ability. This study was supported by the National Research Foundation of Korea and funded by the Korean government.

The impact of long-term water stress on relative growth rate and morphology of needles and shoots of Metasequoia glyptostroboides seedlings: research toward identifying mechanistic models.

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Zhang, Yanxiang; Equiza, Maria Alejandra; Zheng, Quanshui; Tyree, Melvin T

2011-09-01

Leaf morphology in the upper canopy of trees tends to be different from that lower down. The effect of long-term water stress on leaf growth and morphology was studied in seedlings of Metasequoia glyptostroboides to understand how tree height might affect leaf morphology in larger trees. Tree height increases water stress on growing leaves through increased hydraulic resistance to water flow and increased gravitational potential, hence we assume that water stress imposed by soil dehydration will have an effect equivalent to stress induced by height. Seedlings were subjected to well-watered and two constant levels of long-term water stress treatments. Drought treatment significantly reduced final needle count, area and mass per area (leaf mass area, LMA) and increased needle density. Needles from water-stressed plants had lower maximum volumetric elastic modulus (ε(max)), osmotic potential at full turgor (Ψ¹⁰⁰(π)) (and at zero turgor (Ψ⁰(π)) (than those from well-watered plants. Palisade and spongy mesophyll cell size and upper epidermal cell size decreased significantly in drought treatments. Needle relative growth rate, needle length and cell sizes were linear functions of the daily average water potential at the time of leaf growth (r² 0.88-0.999). We conclude that water stress alone does mimic the direction and magnitude of changes in leaf morphology observed in tall trees. The results are discussed in terms of various models for leaf growth rate. Copyright © Physiologia Plantarum 2011.

Anatomia foliar de quatro espécies do gênero Cattleya Lindl. (Orchidaceae do Planalto Central Brasileiro Foliar anatomy of four species of genus Cattleya Lindl. (Orchidaceae of the Brazilian Central Planalt

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Rosane Zanenga-Godoy

2003-03-01

Full Text Available As espécies analisadas (Cattleya araguaiensis Pabst, C. bicolor Lindl., C. nobilior Rchb. e C. walkeriana Gardn. apresentam epiderme uniestratificada em ambas as faces da lâmina foliar, ocorrendo estômatos na face abaxial; deposição de cera epicuticular em crostas, escamas, flocos e plaquetas; em C. araguaiensis ocorrem estegmatas incrustados nas células epidérmicas; em todas as espécies ocorre hipoderme com células de paredes espessadas; mesofilo bifacial, compacto; células paliçádicas atípicas, com barras de espessamento; feixes vasculares colaterais dispostos alternadamente, acompanhados por bainha de fibras; cordões fibrosos acompanhados por estegmatas ocorrem longitudinalmente na lâmina foliar; presença de ráfides. Os aspectos descritos revelam acentuada xeromorfia, em função de economia de água.The analyzed species (Cattleya araguaiensis Pabst, C. bicolor Lindl., C. nobilior Rchb. e C. walkeriana Gardn. present unistratified epidermis at both surfaces with stomata at abaxial surface; deposition of epicuticular wax in crusts, scales, granules and plates; C. araguaiensis presents stegmata at epidermal cells; others characteristics present at all species are: hypodermis cells with thick walls; bifacial and compact mesophyll; atipic cells ofpalisade parenchyma with thichness bars; collateral vascular bundles alternate accompanied by sheath fibers; cordon fibers accompanied by stegmata in the foliar blade; raphids. The described aspects reveal accentuate xeromorphy for water economy.

Ultrastructural and physiological responses of potato (Solanum tuberosum L.) plantlets to gradient saline stress

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Gao, Hui-Juan; Yang, Hong-Yu; Bai, Jiang-Ping; Liang, Xin-Yue; Lou, Yan; Zhang, Jun-Lian; Wang, Di; Zhang, Jin-Lin; Niu, Shu-Qi; Chen, Ying-Long

2015-01-01

Salinity is one of the major abiotic stresses that impacts plant growth and reduces the productivity of field crops. Compared to field plants, test tube plantlets offer a direct and fast approach to investigate the mechanism of salt tolerance. Here we examined the ultrastructural and physiological responses of potato (Solanum tuberosum L. c.v. “Longshu No. 3”) plantlets to gradient saline stress (0, 25, 50, 100, and 200 mM NaCl) with two consequent observations (2 and 6 weeks, respectively). The results showed that, with the increase of external NaCl concentration and the duration of treatments, (1) the number of chloroplasts and cell intercellular spaces markedly decreased, (2) cell walls were thickened and even ruptured, (3) mesophyll cells and chloroplasts were gradually damaged to a complete disorganization containing more starch, (4) leaf Na and Cl contents increased while leaf K content decreased, (5) leaf proline content and the activities of catalase (CAT) and superoxide dismutase (SOD) increased significantly, and (6) leaf malondialdehyde (MDA) content increased significantly and stomatal area and chlorophyll content decline were also detected. Severe salt stress (200 mM NaCl) inhibited plantlet growth. These results indicated that potato plantlets adapt to salt stress to some extent through accumulating osmoprotectants, such as proline, increasing the activities of antioxidant enzymes, such as CAT and SOD. The outcomes of this study provide ultrastructural and physiological insights into characterizing potential damages induced by salt stress for selecting salt-tolerant potato cultivars. PMID:25628634

Morphological, Micoscopical Studies and Elemental analysis of Polygonum Chinese L. and Rhoeo discolor Hance

International Nuclear Information System (INIS)

Moe Moe Lwin

2008-03-01

In the present study, two selected medicinal plants, namely Polygonum chinense L. (Mahaga-Kyansit) and Rhoeo discolor Hance. (Mi-gwin-gamon) are collected from Dawei District. In this paper, the outstanding features of selected plants, microscopical characters and elemental analysis of these leaves are also presented.In morphological studies of Mahaga-kyansit, herbs, swollen at nodes, black-coloured patches present at central portion of leaf blade, ochreate stipules. The inflorescences are terminally corymbose. The ovary trigonous-ovoid, stigma trifixed. In microscopical studies, stomata anomocytic. The cuticle is striation on both surface. In transverse section of midrib, the vascular bundles are collateral, about 8-10 numbers in basal, one was larger than the other bundles, 2numbers in middle regions and 1 bundle in apical regions. Calcium oxalate crystals (Rosette) are present in the parenchymatous cells. In the present study, the Migwin-gamon are rosette-forming succulent herbs. The leaves are crowded, upright, the upper surface green, the lower surface rich reddish-purple. The inflorescences are axillary, boat-shaped. Flowers involucres bracts. The androecium are stamens 6, often clothed wIth moniliform hairs, anthers often with and expanded or variously modified connective. The gynaecium are carpel 3,, trilocular, stigma capitate. In the microscopical studies, the leaves are isobilateral leaf. The stomata are tetracytic type. Lower epidermis cells larger than the upper one and stoma in the lower epidermis. Crystal of tetragonal system form (Prism) present in the mesophyll cells.

Structural and metabolic transitions of C4 leaf development and differentiation defined by microscopy and quantitative proteomics in maize.

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Majeran, Wojciech; Friso, Giulia; Ponnala, Lalit; Connolly, Brian; Huang, Mingshu; Reidel, Edwin; Zhang, Cankui; Asakura, Yukari; Bhuiyan, Nazmul H; Sun, Qi; Turgeon, Robert; van Wijk, Klaas J

2010-11-01

C(4) grasses, such as maize (Zea mays), have high photosynthetic efficiency through combined biochemical and structural adaptations. C(4) photosynthesis is established along the developmental axis of the leaf blade, leading from an undifferentiated leaf base just above the ligule into highly specialized mesophyll cells (MCs) and bundle sheath cells (BSCs) at the tip. To resolve the kinetics of maize leaf development and C(4) differentiation and to obtain a systems-level understanding of maize leaf formation, the accumulation profiles of proteomes of the leaf and the isolated BSCs with their vascular bundle along the developmental gradient were determined using large-scale mass spectrometry. This was complemented by extensive qualitative and quantitative microscopy analysis of structural features (e.g., Kranz anatomy, plasmodesmata, cell wall, and organelles). More than 4300 proteins were identified and functionally annotated. Developmental protein accumulation profiles and hierarchical cluster analysis then determined the kinetics of organelle biogenesis, formation of cellular structures, metabolism, and coexpression patterns. Two main expression clusters were observed, each divided in subclusters, suggesting that a limited number of developmental regulatory networks organize concerted protein accumulation along the leaf gradient. The coexpression with BSC and MC markers provided strong candidates for further analysis of C(4) specialization, in particular transporters and biogenesis factors. Based on the integrated information, we describe five developmental transitions that provide a conceptual and practical template for further analysis. An online protein expression viewer is provided through the Plant Proteome Database.

Profiling of wheat class III peroxidase genes derived from powdery mildew-attacked epidermis reveals distinct sequence-associated expression patterns.

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Liu, Guosheng; Sheng, Xiaoyan; Greenshields, David L; Ogieglo, Adam; Kaminskyj, Susan; Selvaraj, Gopalan; Wei, Yangdou

2005-07-01

A cDNA library was constructed from leaf epidermis of diploid wheat (Triticum monococcum) infected with the powdery mildew fungus (Blumeria graminis f. sp. tritici) and was screened for genes encoding peroxidases. From 2,500 expressed sequence tags (ESTs), 36 cDNAs representing 10 peroxidase genes (designated TmPRX1 to TmPRX10) were isolated and further characterized. Alignment of the deduced amino acid sequences and phylogenetic clustering with peroxidases from other plant species demonstrated that these peroxidases fall into four distinct groups. Differential expression and tissue-specific localization among the members were observed during the B. graminis f. sp. tritici attack using Northern blots and reverse-transcriptase polymerase chain reaction analyses. Consistent with its abundance in the EST collection, TmPRX1 expression showed the highest induction during pathogen attack and fluctuated in response to the fungal parasitic stages. TmPRX1 to TmPRX6 were expressed predominantly in mesophyll cells, whereas TmPRX7 to TmPRX10, which feature a putative C-terminal propeptide, were detectable mainly in epidermal cells. Using TmPRX8 as a representative, we demonstrated that its C-terminal propeptide was sufficient to target a green fluorescent protein fusion protein to the vacuoles in onion cells. Finally, differential expression profiles of the TmPRXs after abiotic stresses and signal molecule treatments were used to dissect the potential role of these peroxidases in multiple stress and defense pathways.

Regulation of cell cycle progression by cell-cell and cell-matrix forces

NARCIS (Netherlands)

Uroz, Marina; Wistorf, Sabrina; Serra-Picamal, Xavier; Conte, Vito; Sales-Pardo, Marta; Roca-Cusachs, Pere; Guimerà, Roger; Trepat, Xavier

2018-01-01

It has long been proposed that the cell cycle is regulated by physical forces at the cell-cell and cell-extracellular matrix (ECM) interfaces 1-12 . However, the evolution of these forces during the cycle has never been measured in a tissue, and whether this evolution affects cell cycle progression

Plastidic Phosphoglucose Isomerase Is an Important Determinant of Starch Accumulation in Mesophyll Cells, Growth, Photosynthetic Capacity, and Biosynthesis of Plastidic Cytokinins in Arabidopsis

Czech Academy of Sciences Publication Activity Database

Bahaji, A.; Sanchez-Lopez, A.M.; De Diego, N.; Munoz, F.J.; Humplík, J.F.; Novák, Ondřej; Spíchal, L.; Doležal, K.; Pozueta-Romero, J.

2015-01-01

Roč. 10, č. 3 (2015) E-ISSN 1932-6203 R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : ADP-GLUCOSE PYROPHOSPHORYLASE * PENTOSE-PHOSPHATE PATHWAY * POSTTRANSLATIONAL REDOX-MODIFICATION Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.057, year: 2015

Well-Controlled Cell-Trapping Systems for Investigating Heterogeneous Cell-Cell Interactions.

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Kamiya, Koki; Abe, Yuta; Inoue, Kosuke; Osaki, Toshihisa; Kawano, Ryuji; Miki, Norihisa; Takeuchi, Shoji

2018-03-01

Microfluidic systems have been developed for patterning single cells to study cell-cell interactions. However, patterning multiple types of cells to understand heterogeneous cell-cell interactions remains difficult. Here, it is aimed to develop a cell-trapping device to assemble multiple types of cells in the well-controlled order and morphology. This device mainly comprises a parylene sheet for assembling cells and a microcomb for controlling the cell-trapping area. The cell-trapping area is controlled by moving the parylene sheet on an SU-8 microcomb using tweezers. Gentle downward flow is used as a driving force for the cell-trapping. The assembly of cells on a parylene sheet with round and line-shaped apertures is demonstrated. The cell-cell contacts of the trapped cells are then investigated by direct cell-cell transfer of calcein via connexin nanopores. Finally, using the device with a system for controlling the cell-trapping area, three different types of cells in the well-controlled order are assembled. The correct cell order rate obtained using the device is 27.9%, which is higher than that obtained without the sliding parylene system (0.74%). Furthermore, the occurrence of cell-cell contact between the three cell types assembled is verified. This cell-patterning device will be a useful tool for investigating heterogeneous cell-cell interactions. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cell Adhesions: Actin-Based Modules that Mediate Cell-Extracellular Matrix and Cell-Cell Interactions

Science.gov (United States)

Bachir, Alexia; Horwitz, Alan Rick; Nelson, W. James; Bianchini, Julie M.

2018-01-01

Cell adhesions link cells to the extracellular matrix (ECM) and to each other, and depend on interactions with the actin cytoskeleton. Both cell-ECM and cell-cell adhesion sites contain discrete, yet overlapping functional modules. These modules establish physical association with the actin cytoskeleton, locally modulate actin organization and dynamics, and trigger intracellular signaling pathways. Interplay between these modules generates distinct actin architectures that underlie different stages, types, and functions of cell-ECM and cell-cell adhesions. Actomyosin contractility is required to generate mature, stable adhesions, as well as sense and translate the mechanical properties of the cellular environment to changes in cell organization and behavior. In this chapter we discuss the organization and function of different adhesion modules and how they interact with the actin cytoskeleton. We highlight the molecular mechanisms of mechanotransduction in adhesions, and how adhesion molecules mediate crosstalk between cell-ECM and cell-cell adhesion sites. PMID:28679638

Mutation induction and isolation in potato through true seed and tuber mutagenesis and use of tissue culture

International Nuclear Information System (INIS)

Upadhya, M.D.; Abraham, M.J.; Dass, B.; Chandra, R.

1982-01-01

Advance MV generation clones from hydrazine-sulphate-treated 'O.T' cultures have been field evaluated and 12 cultures have been selected for yield trials. One culture, DN-31-3, has been found to be day-neutral in its tuberization behaviour. Four JL/RA clones have been selected after a large-scale field trial. These clones are the selections from gamma-irradiated self seeds of Kufri Lauvkar (A-7416) and hybrid A-2235. Similarly 15 MV 3 clones have been selected from the populations raised from EMS- and DES-treated self seeds of A-2235. Day-neutral mutants have been selected from the fourth to seventh sprouts taken from EMS-treated tuber halves of Kufri Jyoti. From the sixth sprout harvest from EMS-treated Kufri Jyoti halves, one mutant, BCN-6-2, has been isolated which showed less than 30 cysts of G. rostochiensis in the MV 2 generation. This clone has been multiplied and made disease-free through apical meristem culture. Through the use of a new medium, PM-32, the plating efficiency of mechanically isolated single callus cells of dihaploid PH-258 is 30-35%. Nitsch's medium has been modified to formulate a new medium for direct embryogenesis in single callus cells of Phulwa. All stages up to the globular stage could be observed after five to six weeks of culture. Efforts were continued for the enzymatic isolation of single-leaf mesophyll cells from dihaploid PH-258. A new cell separation medium has been formulated which gives 80% viable cells. The LD 50 and LD 100 doses of EMS for the single callus cells of dihaploid PH-255 were found to be 500 ppm and 1000 ppm respectively. (author)

Meningeal mast cell-T cell crosstalk regulates T cell encephalitogenicity.

Science.gov (United States)

Russi, Abigail E; Walker-Caulfield, Margaret E; Guo, Yong; Lucchinetti, Claudia F; Brown, Melissa A

2016-09-01

GM-CSF is a cytokine produced by T helper (Th) cells that plays an essential role in orchestrating neuroinflammation in experimental autoimmune encephalomyelitis, a rodent model of multiple sclerosis. Yet where and how Th cells acquire GM-CSF expression is unknown. In this study we identify mast cells in the meninges, tripartite tissues surrounding the brain and spinal cord, as important contributors to antigen-specific Th cell accumulation and GM-CSF expression. In the absence of mast cells, Th cells do not accumulate in the meninges nor produce GM-CSF. Mast cell-T cell co-culture experiments and selective mast cell reconstitution of the meninges of mast cell-deficient mice reveal that resident meningeal mast cells are an early source of caspase-1-dependent IL-1β that licenses Th cells to produce GM-CSF and become encephalitogenic. We also provide evidence of mast cell-T cell co-localization in the meninges and CNS of recently diagnosed acute MS patients indicating similar interactions may occur in human demyelinating disease. Copyright © 2016 Elsevier Ltd. All rights reserved.

Mast cells enhance T cell activation: Importance of mast cell-derived TNF

Science.gov (United States)

Nakae, Susumu; Suto, Hajime; Kakurai, Maki; Sedgwick, Jonathon D.; Tsai, Mindy; Galli, Stephen J.

2005-05-01

Mast cells are not only important effector cells in immediate hypersensitivity reactions and immune responses to pathogens but also can contribute to T cell-mediated disorders. However, the mechanisms by which mast cells might influence T cells in such settings are not fully understood. We find that mast cells can enhance proliferation and cytokine production in multiple T cell subsets. Mast cell-dependent enhancement of T cell activation can be promoted by FcRI-dependent mast cell activation, TNF production by both mast cells and T cells, and mast cell-T cell contact. However, at high concentrations of cells, mast cells can promote T cell activation independent of IgE or TNF. Finally, mast cells also can promote T cell activation by means of soluble factors. These findings identify multiple mechanisms by which mast cells can influence T cell proliferation and cytokine production. allergy | asthma | autoimmunity | cytokines | immune response

Induction of Functional Hair-Cell-Like Cells from Mouse Cochlear Multipotent Cells

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Quanwen Liu

2016-01-01

Full Text Available In this paper, we developed a two-step-induction method of generating functional hair cells from inner ear multipotent cells. Multipotent cells from the inner ear were established and induced initially into progenitor cells committed to the inner ear cell lineage on the poly-L-lysine substratum. Subsequently, the committed progenitor cells were cultured on the mitotically inactivated chicken utricle stromal cells and induced into hair-cell-like cells containing characteristic stereocilia bundles. The hair-cell-like cells exhibited rapid permeation of FM1-43FX. The whole-cell patch-clamp technique was used to measure the membrane currents of cells differentiated for 7 days on chicken utricle stromal cells and analyze the biophysical properties of the hair-cell-like cells by recording membrane properties of cells. The results suggested that the hair-cell-like cells derived from inner ear multipotent cells were functional following differentiation in an enabling environment.

Cell Cycle Related Differentiation of Bone Marrow Cells into Lung Cells

Energy Technology Data Exchange (ETDEWEB)

Dooner, Mark; Aliotta, Jason M.; Pimental, Jeffrey; Dooner, Gerri J.; Abedi, Mehrdad; Colvin, Gerald; Liu, Qin; Weier, Heinz-Ulli; Dooner, Mark S.; Quesenberry, Peter J.

2007-12-31

Green-fluorescent protein (GFP) labeled marrow cells transplanted into lethally irradiated mice can be detected in the lungs of transplanted mice and have been shown to express lung specific proteins while lacking the expression of hematopoietic markers. We have studied marrow cells induced to transit cell cycle by exposure to IL-3, IL-6, IL-11 and steel factor at different times of culture corresponding to different phases of cell cycle. We have found that marrow cells at the G1/S interface have a 3-fold increase in cells which assume a lung phenotype and that this increase is no longer seen in late S/G2. These cells have been characterized as GFP{sup +} CD45{sup -} and GFP{sup +} cytokeratin{sup +}. Thus marrow cells with the capacity to convert into cells with a lung phenotype after transplantation show a reversible increase with cytokine induced cell cycle transit. Previous studies have shown the phenotype of bone marrow stem cells fluctuates reversibly as these cells traverse cell cycle, leading to a continuum model of stem cell regulation. The present studies indicate that marrow stem cell production of nonhematopoietic cells also fluctuates on a continuum.

Deletion of Notch1 converts pro-T cells to dendritic cells and promotes thymic B cells by cell-extrinsic and cell-intrinsic mechanisms.

Science.gov (United States)

Feyerabend, Thorsten B; Terszowski, Grzegorz; Tietz, Annette; Blum, Carmen; Luche, Hervé; Gossler, Achim; Gale, Nicholas W; Radtke, Freddy; Fehling, Hans Jörg; Rodewald, Hans-Reimer

2009-01-16

Notch1 signaling is required for T cell development and has been implicated in fate decisions in the thymus. We showed that Notch1 deletion in progenitor T cells (pro-T cells) revealed their latent developmental potential toward becoming conventional and plasmacytoid dendritic cells. In addition, Notch1 deletion in pro-T cells resulted in large numbers of thymic B cells, previously explained by T-to-B cell fate conversion. Single-cell genotyping showed, however, that the majority of these thymic B cells arose from Notch1-sufficient cells by a cell-extrinsic pathway. Fate switching nevertheless exists for a subset of thymic B cells originating from Notch1-deleted pro-T cells. Chimeric mice lacking the Notch ligand delta-like 4 (Dll4) in thymus epithelium revealed an essential role for Dll4 in T cell development. Thus, Notch1-Dll4 signaling fortifies T cell commitment by suppressing non-T cell lineage potential in pro-T cells, and normal Notch1-driven T cell development repels excessive B cells in the thymus.

Mesenchymal stem cell like (MSCl) cells generated from human embryonic stem cells support pluripotent cell growth

International Nuclear Information System (INIS)

Varga, Nóra; Veréb, Zoltán; Rajnavölgyi, Éva; Német, Katalin; Uher, Ferenc; Sarkadi, Balázs; Apáti, Ágota

2011-01-01

Highlights: ► MSC like cells were derived from hESC by a simple and reproducible method. ► Differentiation and immunosuppressive features of MSCl cells were similar to bmMSC. ► MSCl cells as feeder cells support the undifferentiated growth of hESC. -- Abstract: Mesenchymal stem cell like (MSCl) cells were generated from human embryonic stem cells (hESC) through embryoid body formation, and isolated by adherence to plastic surface. MSCl cell lines could be propagated without changes in morphological or functional characteristics for more than 15 passages. These cells, as well as their fluorescent protein expressing stable derivatives, efficiently supported the growth of undifferentiated human embryonic stem cells as feeder cells. The MSCl cells did not express the embryonic (Oct4, Nanog, ABCG2, PODXL, or SSEA4), or hematopoietic (CD34, CD45, CD14, CD133, HLA-DR) stem cell markers, while were positive for the characteristic cell surface markers of MSCs (CD44, CD73, CD90, CD105). MSCl cells could be differentiated toward osteogenic, chondrogenic or adipogenic directions and exhibited significant inhibition of mitogen-activated lymphocyte proliferation, and thus presented immunosuppressive features. We suggest that cultured MSCl cells can properly model human MSCs and be applied as efficient feeders in hESC cultures.

Involvement of plant stem cells or stem cell-like cells in dedifferentiation

Directory of Open Access Journals (Sweden)

Fangwei eJiang

2015-11-01

Full Text Available Dedifferentiation is the transformation of cells from a given differentiated state to a less differentiated or stem cell-like state. Stem cell-related genes play important roles in dedifferentiation, which exhibits similar histone modification and DNA methylation features to stem cell maintenance. Hence, stem cell-related factors possibly synergistically function to provide a specific niche beneficial to dedifferentiation. During callus formation in Arabidopsis petioles, cells adjacent to procambium cells (stem cell-like cells are dedifferentiated and survive more easily than other cell types. This finding indicates that stem cells or stem cell-like cells may influence the dedifferentiating niche. In this paper, we provide a brief overview of stem cell maintenance and dedifferentiation regulation. We also summarize current knowledge of genetic and epigenetic mechanisms underlying the balance between differentiation and dedifferentiation. Furthermore, we discuss the correlation of stem cells or stem cell-like cells with dedifferentiation.

Morpho-anatomy of the leaf of Myrciaria glomerata

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Nemes Veiga Pacheco-Silva

Full Text Available Abstract Myrciaria glomerata O. Berg., Myrtaceae, popularly known as "cabeludinha", has high content of ascorbic acid and anti-inflammatory property and is used in folk medicine. The objectives of this study were the morphological, anatomical and histochemical characterization of the leaves. Leaf studies were made with optical, scanning electron and confocal microscopy. The collection of botanical material was held at the Tijuca Forest, Rio de Janeiro, RJ. Histochemical tests aimed the identification of lipids, starch grains, phenolic compounds and crystals. The leaves are simple, opposite, lanceolate, pinnate, hairy, with involute margins, hypostomatic and dorsiventral. The stomata are anomocytic. The epidermis presents simple trichomes. Epidermal cells show uneven thickening of their periclinal outer walls, mainly on the adaxial side of the leaf. Secretory cavities of essential oils are subepidermal and exceed, in height, the palisade parenchyma, formed by one cell layer. Four to five cellular layers, rich in phenolic compounds and lipids form the spongy parenchyma. The bundles are collateral and there are many crystals of calcium oxalate spread throughout the mesophyll. In the midrib and petiole the bundles are bicollateral. Analysis by scanning electron revealed epicuticular wax rod-shaped and as grains. In confocal microscopy, the adaxial epidermis, the fibers and the secretory epithelium of the cavities show autofluorescence. The data obtained are important in quality control exams of samples of this species.

Tropical forages: morphoanatomy of plants grown in areas with the death of pasture syndrome

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N. G. Ribeiro-Júnior

2017-05-01

Full Text Available Abstract Roots and leaves of Panicum maximum Tanzânia, Mombaça and Massai; Urochloa brizantha Piatã, Marandu and Xaraés; Urochloa humidicola Llanero; Urochloa ruziziensis Ruzizienses; Urochloa hybrida Mulato II and Cynodon nlemfuensis Estrela-roxa were analyzed, seeking to identify characters for better adaptation to the environment that may interfere with digestibility of tissue from the point of view of the rumen in cattle. Were planted ten cultivars in a completely randomized blocks with three repetitions. Was collected vegetative material, which histological slides were prepared from middle third of the sections of roots and leaves. Were observed differences (p>0.05 in the roots: higher volume of epidermal cells (28.62 µm and overall diameter (1926.41 µm of Llanero; thicker vascular cylinder (975.09 µm and more protoxylem (42.25 in Estrela-roxa and occurrence of aerenchyma in cultivars Piatã, Mulato II, Xaraés, Massai, Llanero and Estrela-roxa; Were found higher proportions of bulliform cells in the leaves (121.07 µm and thicker leaf mesophyll in U. humidicola Llanero (263.63 µm; higher proportion of sclerenchyma fibers in Xaraés and Marandu; lower results for amount of fibers in P. maximum Massai. We conclude that the cultivars Estrela-roxa, Llanero and Massai have greater adaptability to the environment and better nutritional quality.

Cell-cell interactions mediate cytoskeleton organization and collective endothelial cell chemotaxis.

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Shamloo, Amir

2014-09-01

This study investigates the role of cell-cell and cell-ligand interactions in cytoskeleton organization of endothelial cells (ECs) and their directional migration within a microfluidic device. The migration of ECs in response to a biochemical factor was studied. Mathematical analysis of the cell migration pathways and cellular cytoskeleton revealed that directional migration, migration persistence length, migration speed, and cytoskeletal stress fiber alignment can be mediated by the level of cell contacts as well as the presence or absence of a biochemical polarizing factor. It was shown that in the presence of a biochemical polarizing factor, higher cell density and more frequent cell contacts has a reinforcing effect on collective cell chemotaxis. In contrast, in the absence of a polarizing factor, high cell density can decrease or suppress the ability of the cells to migrate. Also, the correlation of actin stress fiber organization and alignment with directional migration of ECs was investigated. It was shown that in the presence of a biochemical polarizing factor, stress fibers within the cytoskeleton of ECs can be significantly aligned parallel to the gradient direction when the cells have higher level of contacts. The results also show that the organization and alignment of actin stress fibers is mediated by cell adhesion junctions during collective cell migration and introduce cell-cell interactions as a key factor during collective cell chemotaxis. © 2014 Wiley Periodicals, Inc.

Mesenchymal stem cell like (MSCl) cells generated from human embryonic stem cells support pluripotent cell growth

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Varga, Nora [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary); Vereb, Zoltan; Rajnavoelgyi, Eva [Department of Immunology, Medical and Health Science Centre, University of Debrecen, Debrecen (Hungary); Nemet, Katalin; Uher, Ferenc; Sarkadi, Balazs [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary); Apati, Agota, E-mail: apati@kkk.org.hu [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary)

2011-10-28

Highlights: Black-Right-Pointing-Pointer MSC like cells were derived from hESC by a simple and reproducible method. Black-Right-Pointing-Pointer Differentiation and immunosuppressive features of MSCl cells were similar to bmMSC. Black-Right-Pointing-Pointer MSCl cells as feeder cells support the undifferentiated growth of hESC. -- Abstract: Mesenchymal stem cell like (MSCl) cells were generated from human embryonic stem cells (hESC) through embryoid body formation, and isolated by adherence to plastic surface. MSCl cell lines could be propagated without changes in morphological or functional characteristics for more than 15 passages. These cells, as well as their fluorescent protein expressing stable derivatives, efficiently supported the growth of undifferentiated human embryonic stem cells as feeder cells. The MSCl cells did not express the embryonic (Oct4, Nanog, ABCG2, PODXL, or SSEA4), or hematopoietic (CD34, CD45, CD14, CD133, HLA-DR) stem cell markers, while were positive for the characteristic cell surface markers of MSCs (CD44, CD73, CD90, CD105). MSCl cells could be differentiated toward osteogenic, chondrogenic or adipogenic directions and exhibited significant inhibition of mitogen-activated lymphocyte proliferation, and thus presented immunosuppressive features. We suggest that cultured MSCl cells can properly model human MSCs and be applied as efficient feeders in hESC cultures.

Seasonal variability of foliar photosynthetic and morphological traits and drought impacts in a Mediterranean mixed forest.

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Sperlich, D; Chang, C T; Peñuelas, J; Gracia, C; Sabaté, S

2015-05-01

The Mediterranean region is a hot spot of climate change vulnerable to increased droughts and heat waves. Scaling carbon fluxes from leaf to landscape levels is particularly challenging under drought conditions. We aimed to improve the mechanistic understanding of the seasonal acclimation of photosynthesis and morphology in sunlit and shaded leaves of four Mediterranean trees (Quercus ilex L., Pinus halepensis Mill., Arbutus unedo L. and Quercus pubescens Willd.) under natural conditions. Vc,max and Jmax were not constant, and mesophyll conductance was not infinite, as assumed in most terrestrial biosphere models, but varied significantly between seasons, tree species and leaf position. Favourable conditions in winter led to photosynthetic recovery and growth in the evergreens. Under moderate drought, adjustments in the photo/biochemistry and stomatal/mesophyllic diffusion behaviour effectively protected the photosynthetic machineries. Severe drought, however, induced early leaf senescence mostly in A. unedo and Q. pubescens, and significantly increased leaf mass per area in Q. ilex and P. halepensis. Shaded leaves had lower photosynthetic potentials but cushioned negative effects during stress periods. Species-specificity, seasonal variations and leaf position are key factors to explain vegetation responses to abiotic stress and hold great potential to reduce uncertainties in terrestrial biosphere models especially under drought conditions. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Comparative Analysis of the Anatomy of Two Populations of Red-Root Amaranth (Amaranthus retroflexus L.

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Sava Vrbničanin

2009-01-01

Full Text Available The anatomy of stems and leaves of two populations of the weed species Amaranthus retroflexus L. (red-root amaranth (pop. AMARE1 having green stems covered in sparse hairs and pop. AMARE2 with green but notably dense stem hairs was analysed in order better to understand the uptake and translocation of herbicides that could be indicative of the species’ evolving resistance to herbicides. Samples of the two populations (AMARE1 and AMARE2 were collected from arable land of the Institute of Maize Research at Zemun Polje in 2006. Sampling was performed at the stage of full vegetative growth of plants.Permanent microscoping preparations were made to measure and analyze elements of the anatomy of stems (stem epidermis, cortex, collenchyma, central cylinder and diameter and leaves (leaf epidermis upper surface and underside, mesophyll, leaf thickness and bundle sheath thickness.Both analysed populations of A. retroflexus, morphologically characterized by different density of stem hairiness, were found to have a typical structure of herbaceous dicots. The stem had three distinctive zones: epidermis, cortex and central cylinder. Amaranth leaves have dorsoventral structure, i.e. their upper surface and underside can be differentiated. The results indicated high and very high significance of differences found in stem anatomy between the two analysed populations, while leaf anatomy was not found to display significant differences other than in mesophyll thickness.

Effect of cycocel on photosynthetic activity and essential oil of fennel (Foeniculum vulgare Mill. under drought stress

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Fatemeh Nouri

2014-12-01

Full Text Available Drought stress is one of the most important and most common environmental stresses that limit plant growth. Photosynthesis is the main determinant of plant growth and its retention ability under environmental stress condition is important for preservation of growth stability. To study the effect of spraying CCC on photosynthesis activity and essential oil content on 'Foeniculum vulgare', an experiment was done in split plot design based on complete block with four replications in research field of University of Zanjan in 2011. Levels of drought stress included, control, soft stress (when 30% of available water was out of soil, severe stress (when 90% of available water was out of soil, and three concentration of CCC 0, 1500 and 3000 mg/L. The results showed that between rates foliar application on physiologic characteristics as photosynthesis rate, transpiration severity, RWC and TΔ intercellular (Ci, stomata conductivity, mesophyll conductivity and essential oil content was significant. By spraying 3000 mg/L CCC, balanced the plant position against drought stress and could reduce negative effect. Foliar application of CCC caused significant increase in photosynthesis rate, mesophyll conductivity and significant decrease stomata conductivity, transpiration severity under drought stress and could increase essential oil content under soft stress. In this study, foliar application of CCC to content 3000 mg/L had the highest effect on this characters.

Diffusive and Metabolic Constraints to Photosynthesis in Quinoa during Drought and Salt Stress

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Dilek Killi

2017-10-01

Full Text Available Quinoa (Chenopodium quinoa Willd. has been proposed as a hardy alternative to traditional grain crops in areas with warm-to-hot climates that are likely to experience increased drought and salt stress in the future. We characterised the diffusive and metabolic limitations to photosynthesis in quinoa exposed to drought and salt stress in isolation and combination. Drought-induced pronounced stomatal and mesophyll limitations to CO2 transport, but quinoa retained photosynthetic capacity and photosystem II (PSII performance. Saline water (300 mmol NaCl-equivalent to 60% of the salinity of sea-water supplied in identical volumes to the irrigation received by the control and drought treatments induced similar reductions in stomatal and mesophyll conductance, but also reduced carboxylation of ribulose-1,5-bisphosphate carboxylase/oxygenase, regeneration of ribulose-1,5-bisphosphate, increased non-photochemical dissipation of energy as heat and impaired PSII electron transport. This suggests that ion toxicity reduced PN via interference with photosynthetic enzymes and degradation of pigment–protein complexes within the thylakoid membranes. The results of this study demonstrate that the photosynthetic physiology of quinoa is resistant to the effects of drought, but quinoa may not be a suitable crop for areas subject to strong salt stress or irrigation with a concentration of saline water equivalent to a 300 mmol NaCl solution.

Leaf size indices and structure of the peat swamp forest

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L.G. Aribal

2017-12-01

Full Text Available Leaf size indices of the tree species in the peatland of Agusan del Sur in Mindanao in Philippines was examined to deduce the variation of forest structure and observed forest zonation.  Using raunkiaer and webb’s leaf size classification, the leaf morphometrics of seven tree species consistently found on the established sampling plots were determined.  The species includes Ternstroemia philippinensis Merr., Polyscias aherniana Merr. Lowry and G.M. Plunkett, Calophyllum sclerophyllum Vesque, Fagraea racemosa Jack, Ilex cymosa Blume, Syzygium tenuirame (Miq. Merr. and Tristaniopsis micrantha Merr. Peter G.Wilson and J.T.Waterh.The LSI were correlated against the variables of the peat physico-chemical properties (such as bulk density, acrotelm thickness, peat depth, total organic carbon, nitrogen, phosphorus, and potassium, pH; water (pH, ammonium, nitrate, phosphate; and leaf tissue elements (nitrogen, phosphorus and potassium.  Result showed a decreasing leaf size indices and a three leaf size category consisting of mesophyllous, mesophyllous-notophyllous and microphyllous were observed which corresponds to the structure of vegetation i.e., from the tall-pole forest having the biggest average leaf area of 6,142.29 mm2 to the pygmy forest with average leaf area of 1,670.10 mm2.  Such decreased leaf size indices were strongly correlated to soil nitrogen, acrotelm thickness, peat depth, phosphate in water, nitrogen and phosphorus in the plant tissue.

ECOLOGICAL AND FAUNISTIC REVIEW OF COCCINELLIDAE (COLEOPTERA, COCCINELLIDAE IN THE REPUBLIC OF DAGESTAN

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G. M. Mukhtarova

2016-01-01

Full Text Available Aim. The aim of the study is to conduct ecological and faunal studies of coccinellidae in Dagestan as well as to consider the chorologic, trophic and phenological aspects in connection with the peculiarities of the environment.Materials and methods. As the basis for the research we used the materials obtained by the authors from 1999 to 2016 in various parts of Dagestan. We also conducted expeditions, stationary and semi-stationary studies using traditional methods of entomological research.Results. Inventory of coccinellidae fauna in Dagestan allowed identifying 27 species from 18 genera. It also revealed multiple and rare species. We carried out an analysis on distribution of the species in the high altitude range; identified environmental groups by biotopic preferendum: xerophyllous, mesophyll meadow-steppe, mesophyll forest, and Hygrophila and polytopic species; according to trophic adaptation: Aphidophages, Coccidophages, Mycetophages, Entomophages and Phytophages; We held a zoogeographical analysis of coccinellidae of Dagestan and according to habitat type, 8 zoogeographic groups were identified; We conducted phenological observations of the study sites, stages and timing of winter and winter dormancy, the beginning of mating, egg-laying, the duration of certain stages, the number of generations and other features.Conclusion. This research represents the first comprehensive summary of ladybugs (Coleoptera, Coccinellidae of Dagestan and includes the findings of the composition study, ecological and zoogeographical aspects of coccinellidae fauna of the study area.

LEAF MICROMORPHOMETRY OF Schinus molle L. (ANARCADIACEAE IN DIFFERENT CANOPY HEIGHTS.

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Marinês Ferreira Pires

2015-03-01

Full Text Available Leaf characterization of trees is essential for its identification and use, as well as to understand its relationships with environment. The objective of this work is to study the leaflet anatomy and leaf biometrical characteristics at different canopy heights of Schinus molle plants as a function of its environmental and physiological modifications. Leaves were collected at three different canopy heights: base, middle and upper canopy in a plantation of S. molle. Leaves were used for anatomical and biometrical analysis. For the anatomical analysis, leaves were fixed in FAA and stored in ethanol 70% and further submitted to transversal and paradermical sections. Slides were photomicrographed and image analysis was performed in UTHSCSA-Imagetool. For biometrical analysis leaf area, length, width, dry mass and specific leaf area were evaluated. The leaflets exhibited single layer epidermis, anomocytic and ciclocytic stomata, isobilateral mesophyll, subepidermal parenchyma layer in both adaxial and abaxial faces of epidermis, secretory vessels and lamellar collenchyma in midrib and leaf border. Leaf anatomy modifications occurred in cuticle and mesophyll thickness, vascular system, phloem thickness, and stomatal density in accordance with leaf canopy position. Leaves were smaller and with reduced leaf area at higher canopy positions. S. molle leaf anatomy is different from other species within Schinus genre with modifications under different environmental and physiological modifications promoted by its canopy height.

Diffusive and Metabolic Constraints to Photosynthesis in Quinoa during Drought and Salt Stress

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Killi, Dilek; Haworth, Matthew

2017-01-01

Quinoa (Chenopodium quinoa Willd.) has been proposed as a hardy alternative to traditional grain crops in areas with warm-to-hot climates that are likely to experience increased drought and salt stress in the future. We characterised the diffusive and metabolic limitations to photosynthesis in quinoa exposed to drought and salt stress in isolation and combination. Drought-induced pronounced stomatal and mesophyll limitations to CO2 transport, but quinoa retained photosynthetic capacity and photosystem II (PSII) performance. Saline water (300 mmol NaCl-equivalent to 60% of the salinity of sea-water) supplied in identical volumes to the irrigation received by the control and drought treatments induced similar reductions in stomatal and mesophyll conductance, but also reduced carboxylation of ribulose-1,5-bisphosphate carboxylase/oxygenase, regeneration of ribulose-1,5-bisphosphate, increased non-photochemical dissipation of energy as heat and impaired PSII electron transport. This suggests that ion toxicity reduced PN via interference with photosynthetic enzymes and degradation of pigment–protein complexes within the thylakoid membranes. The results of this study demonstrate that the photosynthetic physiology of quinoa is resistant to the effects of drought, but quinoa may not be a suitable crop for areas subject to strong salt stress or irrigation with a concentration of saline water equivalent to a 300 mmol NaCl solution. PMID:29039809

Computational cell model based on autonomous cell movement regulated by cell-cell signalling successfully recapitulates the "inside and outside" pattern of cell sorting

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Ajioka Itsuki

2007-09-01

Full Text Available Abstract Background Development of multicellular organisms proceeds from a single fertilized egg as the combined effect of countless numbers of cellular interactions among highly dynamic cells. Since at least a reminiscent pattern of morphogenesis can be recapitulated in a reproducible manner in reaggregation cultures of dissociated embryonic cells, which is known as cell sorting, the cells themselves must possess some autonomous cell behaviors that assure specific and reproducible self-organization. Understanding of this self-organized dynamics of heterogeneous cell population seems to require some novel approaches so that the approaches bridge a gap between molecular events and morphogenesis in developmental and cell biology. A conceptual cell model in a computer may answer that purpose. We constructed a dynamical cell model based on autonomous cell behaviors, including cell shape, growth, division, adhesion, transformation, and motility as well as cell-cell signaling. The model gives some insights about what cellular behaviors make an appropriate global pattern of the cell population. Results We applied the model to "inside and outside" pattern of cell-sorting, in which two different embryonic cell types within a randomly mixed aggregate are sorted so that one cell type tends to gather in the central region of the aggregate and the other cell type surrounds the first cell type. Our model can modify the above cell behaviors by varying parameters related to them. We explored various parameter sets with which the "inside and outside" pattern could be achieved. The simulation results suggested that direction of cell movement responding to its neighborhood and the cell's mobility are important for this specific rearrangement. Conclusion We constructed an in silico cell model that mimics autonomous cell behaviors and applied it to cell sorting, which is a simple and appropriate phenomenon exhibiting self-organization of cell population. The model

Adaptation of European beech (Fagus silvatica L.) to different ecological conditions: leaf size variation

International Nuclear Information System (INIS)

Barna, M.

2004-01-01

In beech trees, both leaf morphology and leaf area show considerable adaptation capabilities to the local radiation climate. The plants adapting to shade conditions create large leaf area with high chlorophyll concentration and high water content in the living tissues. On the other hand, the leaves of plants exposed to radiation of higher intensity have smaller area, several layers of mesophyll, thick epidermis and cuticle, higher dry weight, higher energy potential of the dry mass and several other characteristic properties

A Common histone modification code on C4 genes in maize and its conservation in Sorghum and Setaria italica.

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Heimann, Louisa; Horst, Ina; Perduns, Renke; Dreesen, Björn; Offermann, Sascha; Peterhansel, Christoph

2013-05-01

C4 photosynthesis evolved more than 60 times independently in different plant lineages. Each time, multiple genes were recruited into C4 metabolism. The corresponding promoters acquired new regulatory features such as high expression, light induction, or cell type-specific expression in mesophyll or bundle sheath cells. We have previously shown that histone modifications contribute to the regulation of the model C4 phosphoenolpyruvate carboxylase (C4-Pepc) promoter in maize (Zea mays). We here tested the light- and cell type-specific responses of three selected histone acetylations and two histone methylations on five additional C4 genes (C4-Ca, C4-Ppdk, C4-Me, C4-Pepck, and C4-RbcS2) in maize. Histone acetylation and nucleosome occupancy assays indicated extended promoter regions with regulatory upstream regions more than 1,000 bp from the transcription initiation site for most of these genes. Despite any detectable homology of the promoters on the primary sequence level, histone modification patterns were highly coregulated. Specifically, H3K9ac was regulated by illumination, whereas H3K4me3 was regulated in a cell type-specific manner. We further compared histone modifications on the C4-Pepc and C4-Me genes from maize and the homologous genes from sorghum (Sorghum bicolor) and Setaria italica. Whereas sorghum and maize share a common C4 origin, C4 metabolism evolved independently in S. italica. The distribution of histone modifications over the promoters differed between the species, but differential regulation of light-induced histone acetylation and cell type-specific histone methylation were evident in all three species. We propose that a preexisting histone code was recruited into C4 promoter control during the evolution of C4 metabolism.

A Common Histone Modification Code on C4 Genes in Maize and Its Conservation in Sorghum and Setaria italica1[W][OA

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Heimann, Louisa; Horst, Ina; Perduns, Renke; Dreesen, Björn; Offermann, Sascha; Peterhansel, Christoph

2013-01-01

C4 photosynthesis evolved more than 60 times independently in different plant lineages. Each time, multiple genes were recruited into C4 metabolism. The corresponding promoters acquired new regulatory features such as high expression, light induction, or cell type-specific expression in mesophyll or bundle sheath cells. We have previously shown that histone modifications contribute to the regulation of the model C4 phosphoenolpyruvate carboxylase (C4-Pepc) promoter in maize (Zea mays). We here tested the light- and cell type-specific responses of three selected histone acetylations and two histone methylations on five additional C4 genes (C4-Ca, C4-Ppdk, C4-Me, C4-Pepck, and C4-RbcS2) in maize. Histone acetylation and nucleosome occupancy assays indicated extended promoter regions with regulatory upstream regions more than 1,000 bp from the transcription initiation site for most of these genes. Despite any detectable homology of the promoters on the primary sequence level, histone modification patterns were highly coregulated. Specifically, H3K9ac was regulated by illumination, whereas H3K4me3 was regulated in a cell type-specific manner. We further compared histone modifications on the C4-Pepc and C4-Me genes from maize and the homologous genes from sorghum (Sorghum bicolor) and Setaria italica. Whereas sorghum and maize share a common C4 origin, C4 metabolism evolved independently in S. italica. The distribution of histone modifications over the promoters differed between the species, but differential regulation of light-induced histone acetylation and cell type-specific histone methylation were evident in all three species. We propose that a preexisting histone code was recruited into C4 promoter control during the evolution of C4 metabolism. PMID:23564230

Endothelial cell subpopulations in vitro: cell volume, cell cycle, and radiosensitivity

International Nuclear Information System (INIS)

Rubin, D.B.; Drab, E.A.; Bauer, K.D.

1989-01-01

Vascular endothelial cells (EC) are important clinical targets of radiation and other forms of free radical/oxidant stresses. In this study, we found that the extent of endothelial damage may be determined by the different cytotoxic responses of EC subpopulations. The following characteristics of EC subpopulations were examined: (1) cell volume; (2) cell cycle position; and (3) cytotoxic indexes for both acute cell survival and proliferative capacity after irradiation (137Cs, gamma, 0-10 Gy). EC cultured from bovine aortas were separated by centrifugal elutriation into subpopulations of different cell volumes. Through flow cytometry, we found that cell volume was related to the cell cycle phase distribution. The smallest EC were distributed in G1 phase and the larger cells were distributed in either early S, middle S, or late S + G2M phases. Cell cycle phase at the time of irradiation was not associated with acute cell loss. However, distribution in the cell cycle did relate to cell survival based on proliferative capacity (P less than 0.01). The order of increasing radioresistance was cells in G1 (D0 = 110 cGy), early S (135 cGy), middle S (145 cGy), and late S + G2M phases (180 cGy). These findings (1) suggest an age-related response to radiation in a nonmalignant differentiated cell type and (2) demonstrate EC subpopulations in culture

Quantitative imaging of epithelial cell scattering identifies specific inhibitors of cell motility and cell-cell dissociation

NARCIS (Netherlands)

Loerke, D.; le Duc, Q.; Blonk, I.; Kerstens, A.; Spanjaard, E.; Machacek, M.; Danuser, G.; de Rooij, J.

2012-01-01

The scattering of cultured epithelial cells in response to hepatocyte growth factor (HGF) is a model system that recapitulates key features of metastatic cell behavior in vitro, including disruption of cell-cell adhesions and induction of cell migration. We have developed image analysis tools that

The cell cycle as a brake for β-cell regeneration from embryonic stem cells.

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El-Badawy, Ahmed; El-Badri, Nagwa

2016-01-13

The generation of insulin-producing β cells from stem cells in vitro provides a promising source of cells for cell transplantation therapy in diabetes. However, insulin-producing cells generated from human stem cells show deficiency in many functional characteristics compared with pancreatic β cells. Recent reports have shown molecular ties between the cell cycle and the differentiation mechanism of embryonic stem (ES) cells, assuming that cell fate decisions are controlled by the cell cycle machinery. Both β cells and ES cells possess unique cell cycle machinery yet with significant contrasts. In this review, we compare the cell cycle control mechanisms in both ES cells and β cells, and highlight the fundamental differences between pluripotent cells of embryonic origin and differentiated β cells. Through critical analysis of the differences of the cell cycle between these two cell types, we propose that the cell cycle of ES cells may act as a brake for β-cell regeneration. Based on these differences, we discuss the potential of modulating the cell cycle of ES cells for the large-scale generation of functionally mature β cells in vitro. Further understanding of the factors that modulate the ES cell cycle will lead to new approaches to enhance the production of functional mature insulin-producing cells, and yield a reliable system to generate bona fide β cells in vitro.

Characterization and mapping of a novel light-dependent lesion mimic mutant Imm6 in rice （Oryza sativa L.）

Institute of Scientific and Technical Information of China (English)

XIAO Gui-qing[1,2; ZHANG Hal-wen[3; LU Xiang-yang[1,2; HUANG Rong-feng[3

2015-01-01

A novel rice lesion mimic mutant （LMM） was isolated from an ethane methyl sulfonate （EMS）-induced 02428 mutant bank. The mutant, tentatively designated as Imm6, develops necrotic lesions in the whole growth period along with changes in several important agronomic traits. We found that the initiation of the lesions was induced by light and cell death occurred in Imm6 accompanied with accumulation of reactive oxygen species （ROS）. The lower chlorophyll content, soluble protein content and superoxide dismutase （SOD） activity, the higher malondialdehyde （MDA） content were detected in Imm6 than in the wild type （WT）. Moreover, the observation by transmission electronic microscope （TEM） demonstrated that some organelles were damaged and the stroma lamella of chloroplast was irregular and loose in mesophyll cell of Imm6. In addition, Imm6 was more resistant than WT to rice blast fungus Magnaporthe grisea infection, which was consistent with increased expression of four genes involved in the defense-related reaction. Genetic analysis showed that mutant trait of Imm6 is inherited as a monogenic recessive nuclear gene located on the long arm of chromosome 6. Using simple sequence repeat （SSR） markers, the target gene was finally delimited to an interval of 80.8 kb between markers MM2359 and MM2370, containing 7 annotated genes. Taken together, our results provide the information to identify a new gene involved in rice lesion mimic, which will be helpful in clarifying the mechanism of cell death and disease resistance in rice.

A rate equation model of stomatal responses to vapour pressure deficit and drought

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Shanahan ST

2002-08-01

Full Text Available Abstract Background Stomata respond to vapour pressure deficit (D – when D increases, stomata begin to close. Closure is the result of a decline in guard cell turgor, but the link between D and turgor is poorly understood. We describe a model for stomatal responses to increasing D based upon cellular water relations. The model also incorporates impacts of increasing levels of water stress upon stomatal responses to increasing D. Results The model successfully mimics the three phases of stomatal responses to D and also reproduces the impact of increasing plant water deficit upon stomatal responses to increasing D. As water stress developed, stomata regulated transpiration at ever decreasing values of D. Thus, stomatal sensitivity to D increased with increasing water stress. Predictions from the model concerning the impact of changes in cuticular transpiration upon stomatal responses to increasing D are shown to conform to experimental data. Sensitivity analyses of stomatal responses to various parameters of the model show that leaf thickness, the fraction of leaf volume that is air-space, and the fraction of mesophyll cell wall in contact with air have little impact upon behaviour of the model. In contrast, changes in cuticular conductance and membrane hydraulic conductivity have significant impacts upon model behaviour. Conclusion Cuticular transpiration is an important feature of stomatal responses to D and is the cause of the 3 phase response to D. Feed-forward behaviour of stomata does not explain stomatal responses to D as feedback, involving water loss from guard cells, can explain these responses.

Ultrastructural and physiological responses of potato (Solanum tuberosum L. plantlets to gradient saline stress

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Hui-Juan eGao

2015-01-01

Full Text Available Salinity is one of the major abiotic stresses that impacts plant growth and reduces the productivity of field crops. Compared to field plants, test tube plantlets offer a direct and fast approach to investigate the mechanism of salt tolerance. Here we examined the ultrastructural and physiological responses of potato (Solanum tuberosum L. c.v. ‘Longshu No. 3’ plantlets to gradient saline stress (0, 25, 50, 100 and 200 mM NaCl with two consequent observations (two and six weeks, respectively. The results showed that, with the increase of external NaCl concentration and the duration of treatments, (1 the number of chloroplasts and cell intercellular spaces markedly decreased, (2 cell walls were thickened and even ruptured, (3 mesophyll cells and chloroplasts were gradually damaged to a complete disorganization containing more starch, (4 leaf Na and Cl contents increased while leaf K content decreased, (5 leaf proline content and the activities of catalase (CAT and superoxide dismutase (SOD increased significantly, and (6 leaf malondialdehyde (MDA content increased significantly and stomatal area and chlorophyll content decline were also detected. Severe salt stress (200 mM NaCl inhibited plantlet growth. These results indicated that potato plantlets adapt to salt stress to some extent through accumulating osmoprotectants, such as proline, increasing the activities of antioxidant enzymes, such as CAT and SOD. The outcomes of this study provide ultrastructural and physiological insights into characterizing potential damages induced by salt stress for selecting salt-tolerant potato cultivars.

Pancreatic stellate cells enhance stem cell-like phenotypes in pancreatic cancer cells

International Nuclear Information System (INIS)

Hamada, Shin; Masamune, Atsushi; Takikawa, Tetsuya; Suzuki, Noriaki; Kikuta, Kazuhiro; Hirota, Morihisa; Hamada, Hirofumi; Kobune, Masayoshi; Satoh, Kennichi; Shimosegawa, Tooru

2012-01-01

Highlights: ► Pancreatic stellate cells (PSCs) promote the progression of pancreatic cancer. ► Pancreatic cancer cells co-cultured with PSCs showed enhanced spheroid formation. ► Expression of stem cell-related genes ABCG2, Nestin and LIN28 was increased. ► Co-injection of PSCs enhanced tumorigenicity of pancreatic cancer cells in vivo. ► This study suggested a novel role of PSCs as a part of the cancer stem cell niche. -- Abstract: The interaction between pancreatic cancer cells and pancreatic stellate cells (PSCs), a major profibrogenic cell type in the pancreas, is receiving increasing attention. There is accumulating evidence that PSCs promote the progression of pancreatic cancer by increasing cancer cell proliferation and invasion as well as by protecting them from radiation- and gemcitabine-induced apoptosis. Recent studies have identified that a portion of cancer cells, called “cancer stem cells”, within the entire cancer tissue harbor highly tumorigenic and chemo-resistant phenotypes, which lead to the recurrence after surgery or re-growth of the tumor. The mechanisms that maintain the “stemness” of these cells remain largely unknown. We hypothesized that PSCs might enhance the cancer stem cell-like phenotypes in pancreatic cancer cells. Indirect co-culture of pancreatic cancer cells with PSCs enhanced the spheroid-forming ability of cancer cells and induced the expression of cancer stem cell-related genes ABCG2, Nestin and LIN28. In addition, co-injection of PSCs enhanced tumorigenicity of pancreatic cancer cells in vivo. These results suggested a novel role of PSCs as a part of the cancer stem cell niche.

In silico characterization of cell-cell interactions using a cellular automata model of cell culture.

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Kihara, Takanori; Kashitani, Kosuke; Miyake, Jun

2017-07-14

Cell proliferation is a key characteristic of eukaryotic cells. During cell proliferation, cells interact with each other. In this study, we developed a cellular automata model to estimate cell-cell interactions using experimentally obtained images of cultured cells. We used four types of cells; HeLa cells, human osteosarcoma (HOS) cells, rat mesenchymal stem cells (MSCs), and rat smooth muscle A7r5 cells. These cells were cultured and stained daily. The obtained cell images were binarized and clipped into squares containing about 10 4 cells. These cells showed characteristic cell proliferation patterns. The growth curves of these cells were generated from the cell proliferation images and we determined the doubling time of these cells from the growth curves. We developed a simple cellular automata system with an easily accessible graphical user interface. This system has five variable parameters, namely, initial cell number, doubling time, motility, cell-cell adhesion, and cell-cell contact inhibition (of proliferation). Within these parameters, we obtained initial cell numbers and doubling times experimentally. We set the motility at a constant value because the effect of the parameter for our simulation was restricted. Therefore, we simulated cell proliferation behavior with cell-cell adhesion and cell-cell contact inhibition as variables. By comparing growth curves and proliferation cell images, we succeeded in determining the cell-cell interaction properties of each cell. Simulated HeLa and HOS cells exhibited low cell-cell adhesion and weak cell-cell contact inhibition. Simulated MSCs exhibited high cell-cell adhesion and positive cell-cell contact inhibition. Simulated A7r5 cells exhibited low cell-cell adhesion and strong cell-cell contact inhibition. These simulated results correlated with the experimental growth curves and proliferation images. Our simulation approach is an easy method for evaluating the cell-cell interaction properties of cells.

Host cell reactivation in mammalian cells

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Lytle, C.D.; Benane, S.G.; Stafford, J.E.

1976-01-01

The survival of UV-irradiated herpes simplex virus was determined in cultured Potoroo (a marsupial) and human cells under lighting conditions which promoted photereactivation. Photoreactivation was readily demonstrated for herpes virus in two lines of Potoroo cells with dose reduction factors of 0.7 to 0.8 for ovary cells and 0.5 to 0.7 for kidney cells. Light from Blacklite (near UV) lamps was more effective than from Daylight (mostly visible) lamps, suggesting that near UV radiation was more effecient for photoreactivation in Potoroo cells. The quantitative and qualitative aspects of this photoreactivation were similar to those reported for a similar virus infecting chick embryo cells. UV-survival curves of herpes virus in Potoroo cells indicated a high level of 'dark' host cell reactivation. No photoreactivation was found for UV-irradiated vaccinia virus in Potoroo cells. A similar photoreactivation study was done using special control lighting (lambda>600 nm) and human cells with normal repair and with cells deficient in excision repair (XP). No photoreactivation was found for UV-irradiated herpes virus in either human cell with either Blacklite or Daylight lamps as the sources of photoreactivating light. This result contrasts with a report of photoreactivation for a herpes virus in the same XP cells using incandescent lamps. (author)

Local cell metrics: a novel method for analysis of cell-cell interactions

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Chen Chien-Chiang

2009-10-01

Full Text Available Abstract Background The regulation of many cell functions is inherently linked to cell-cell contact interactions. However, effects of contact interactions among adherent cells can be difficult to detect with global summary statistics due to the localized nature and noise inherent to cell-cell interactions. The lack of informatics approaches specific for detecting cell-cell interactions is a limitation in the analysis of large sets of cell image data, including traditional and combinatorial or high-throughput studies. Here we introduce a novel histogram-based data analysis strategy, termed local cell metrics (LCMs, which addresses this shortcoming. Results The new LCM method is demonstrated via a study of contact inhibition of proliferation of MC3T3-E1 osteoblasts. We describe how LCMs can be used to quantify the local environment of cells and how LCMs are decomposed mathematically into metrics specific to each cell type in a culture, e.g., differently-labelled cells in fluorescence imaging. Using this approach, a quantitative, probabilistic description of the contact inhibition effects in MC3T3-E1 cultures has been achieved. We also show how LCMs are related to the naïve Bayes model. Namely, LCMs are Bayes class-conditional probability functions, suggesting their use for data mining and classification. Conclusion LCMs are successful in robust detection of cell contact inhibition in situations where conventional global statistics fail to do so. The noise due to the random features of cell behavior was suppressed significantly as a result of the focus on local distances, providing sensitive detection of cell-cell contact effects. The methodology can be extended to any quantifiable feature that can be obtained from imaging of cell cultures or tissue samples, including optical, fluorescent, and confocal microscopy. This approach may prove useful in interpreting culture and histological data in fields where cell-cell interactions play a critical

Local cell metrics: a novel method for analysis of cell-cell interactions.

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Su, Jing; Zapata, Pedro J; Chen, Chien-Chiang; Meredith, J Carson

2009-10-23

The regulation of many cell functions is inherently linked to cell-cell contact interactions. However, effects of contact interactions among adherent cells can be difficult to detect with global summary statistics due to the localized nature and noise inherent to cell-cell interactions. The lack of informatics approaches specific for detecting cell-cell interactions is a limitation in the analysis of large sets of cell image data, including traditional and combinatorial or high-throughput studies. Here we introduce a novel histogram-based data analysis strategy, termed local cell metrics (LCMs), which addresses this shortcoming. The new LCM method is demonstrated via a study of contact inhibition of proliferation of MC3T3-E1 osteoblasts. We describe how LCMs can be used to quantify the local environment of cells and how LCMs are decomposed mathematically into metrics specific to each cell type in a culture, e.g., differently-labelled cells in fluorescence imaging. Using this approach, a quantitative, probabilistic description of the contact inhibition effects in MC3T3-E1 cultures has been achieved. We also show how LCMs are related to the naïve Bayes model. Namely, LCMs are Bayes class-conditional probability functions, suggesting their use for data mining and classification. LCMs are successful in robust detection of cell contact inhibition in situations where conventional global statistics fail to do so. The noise due to the random features of cell behavior was suppressed significantly as a result of the focus on local distances, providing sensitive detection of cell-cell contact effects. The methodology can be extended to any quantifiable feature that can be obtained from imaging of cell cultures or tissue samples, including optical, fluorescent, and confocal microscopy. This approach may prove useful in interpreting culture and histological data in fields where cell-cell interactions play a critical role in determining cell fate, e.g., cancer, developmental

Phloem loading--not metaphysical, only complex: towards a unified model of phloem loading.

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Komor, E; Orlich, G; Weig, A; Köckenberger, W

1996-08-01

Phloem loading comprises the entire pathway of phloem-mobile solutes from their place of generation (or delivery) to the sieve tubes in a sequence of transport steps across or passing by several different cell types. Each of these steps can be classified as symplastic or apoplastic. The detailed anatomical-cytological work in the past ten years made clear that the symplastic continuity from mesophyll to sieve tubes may be very different for different plant species or even in different vein orders. Therefore data from one species are not transferable to another species and a well-rounded picture involving different experimental methods has to be aimed at for each species separately. The information obtained with the Ricinus seedling, where phloem loading and sieve tube sap analysis can be achieved relatively easily, is presented. The analysis of the radioactive labelling of sucrose from the sieve tubes of cotyledons, in which external and intracellular sucrose had been differently labelled, revealed that at sucrose concentrations close to the natural one, 50% of sucrose is loaded directly from the external medium. The other 50% is first taken up by mesophyll and then released for uptake into the sieve tubes. No bundle tissue works as obligate, intermediate sucrose storage. The apoplast therefore definitely serves as a transit reservoir for sucrose destined to be loaded into the sieve tubes. The sieve tube sap contains glycolytic metabolites at concentrations higher than found in the hypocotyl tissue, whereas the corresponding glycolytic enzymes are missing. It is concluded that the enzymes are sequestered in the companion cell or by parietal membrane stacks. Not only the sieve tubes but nearly all cotyledonary cells are equipped with a sucrose-H(+) symporter able to achieve sucrose accumulation and sensitive to inhibition by high salt concentrations or SH reagents. A cDNA clone coding for a sucrose carrier was isolated. It is transcribed at approximately the same

Single-cell sequencing in stem cell biology.

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Wen, Lu; Tang, Fuchou

2016-04-15

Cell-to-cell variation and heterogeneity are fundamental and intrinsic characteristics of stem cell populations, but these differences are masked when bulk cells are used for omic analysis. Single-cell sequencing technologies serve as powerful tools to dissect cellular heterogeneity comprehensively and to identify distinct phenotypic cell types, even within a 'homogeneous' stem cell population. These technologies, including single-cell genome, epigenome, and transcriptome sequencing technologies, have been developing rapidly in recent years. The application of these methods to different types of stem cells, including pluripotent stem cells and tissue-specific stem cells, has led to exciting new findings in the stem cell field. In this review, we discuss the recent progress as well as future perspectives in the methodologies and applications of single-cell omic sequencing technologies.

In situ variation in leaf anatomy and morphology of Andira legalis (Leguminosae in two neighbouring but contrasting light environments in a Brazilian sandy coastal plain Variação in situ em anatomia e morfologia foliar de Andira legalis (Leguminosae em dois ambientes adjacentes, porém contrastantes quanto ao regime de luz, em restinga brasileira

Directory of Open Access Journals (Sweden)

Daniela Carvalho Pereira

2009-03-01

Full Text Available Andira legalis (Vell. Toledo is a legume shrub widespread along the sandy plains of the Brazilian coast. It occurs both shaded, in forest habitats, or exposed to full sunlight, in the vegetation islands growing on sand deposits. Previous studies reported a high range of morpho-physiological variation for this species along a geographical gradient. This study compared leaf morphology and anatomy of A. legalis in two distinct but adjacent light environments: a dense forest (shaded and a scrub of Palmae (exposed. We studied the amplitude of variation for these traits within a small (0.5 ha geographical area. Leaf anatomy parameters were measured for five leaves collected from five plants in each habitat. The parameters measured were leaf and mesophyll thickness, thickness of the outer periclinal cell wall, thickness of the adaxial and abaxial epidermis and vascular bundle transversal section area, and also common epidermal cells, stomata and trichome density. Leaf morphology parameters were obtained from five leaves of each of 20 plants in each site. Dry and fresh weights were measured to obtain leaf specific mass and succulence. All anatomy and morphology parameters, except trichome density, were significantly higher for the sun-exposed plants. Less expected, however, was the marked qualitative difference between exposed and shaded plants: in the former the mesophyll had a unilateral symmetry (i.e., the whole mesophyll occupied by photosynthetic tissue, whereas in the latter there was a dorsiventral symmetry (i.e., partly palisade and partly spongy parenchyma. Such amplitude of variation shows that even within a small geographic area A. legalis has a broad ecological plasticity.Andira legalis (Vell. Toledo é uma leguminosa arbustiva distribuída ao longo de planícies arenosas da costa brasileira. Tem ocorrência em ambientes florestais, sombreadas, ou em ilhas de vegetação de restingas abertas, onde é exposta à plena radiação solar

NKT Cell Responses to B Cell Lymphoma.

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Li, Junxin; Sun, Wenji; Subrahmanyam, Priyanka B; Page, Carly; Younger, Kenisha M; Tiper, Irina V; Frieman, Matthew; Kimball, Amy S; Webb, Tonya J

2014-06-01

Natural killer T (NKT) cells are a unique subset of CD1d-restricted T lymphocytes that express characteristics of both T cells and natural killer cells. NKT cells mediate tumor immune-surveillance; however, NKT cells are numerically reduced and functionally impaired in lymphoma patients. Many hematologic malignancies express CD1d molecules and co-stimulatory proteins needed to induce anti-tumor immunity by NKT cells, yet most tumors are poorly immunogenic. In this study, we sought to investigate NKT cell responses to B cell lymphoma. In the presence of exogenous antigen, both mouse and human NKT cell lines produce cytokines following stimulation by B cell lymphoma lines. NKT cell populations were examined ex vivo in mouse models of spontaneous B cell lymphoma, and it was found that during early stages, NKT cell responses were enhanced in lymphoma-bearing animals compared to disease-free animals. In contrast, in lymphoma-bearing animals with splenomegaly and lymphadenopathy, NKT cells were functionally impaired. In a mouse model of blastoid variant mantle cell lymphoma, treatment of tumor-bearing mice with a potent NKT cell agonist, α-galactosylceramide (α-GalCer), resulted in a significant decrease in disease pathology. Ex vivo studies demonstrated that NKT cells from α-GalCer treated mice produced IFN-γ following α-GalCer restimulation, unlike NKT cells from vehicle-control treated mice. These data demonstrate an important role for NKT cells in the immune response to an aggressive hematologic malignancy like mantle cell lymphoma.

Automated Cell-Cutting for Cell Cloning

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Ichikawa, Akihiko; Tanikawa, Tamio; Matsukawa, Kazutsugu; Takahashi, Seiya; Ohba, Kohtaro

We develop an automated cell-cutting technique for cell cloning. Animal cells softened by the cytochalasin treatment are injected into a microfluidic chip. The microfluidic chip contains two orthogonal channels: one microchannel is wide, used to transport cells, and generates the cutting flow; the other is thin and used for aspiration, fixing, and stretching of the cell. The injected cell is aspirated and stretched in the thin microchannel. Simultaneously, the volumes of the cell before and after aspiration are calculated; the volumes are used to calculate the fluid flow required to aspirate half the volume of the cell into the thin microchannel. Finally, we apply a high-speed flow in the orthogonal microchannel to bisect the cell. This paper reports the cutting process, the cutting system, and the results of the experiment.

Different cell fates from cell-cell interactions: core architectures of two-cell bistable networks.

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Rouault, Hervé; Hakim, Vincent

2012-02-08

The acquisition of different fates by cells that are initially in the same state is central to development. Here, we investigate the possible structures of bistable genetic networks that can allow two identical cells to acquire different fates through cell-cell interactions. Cell-autonomous bistable networks have been previously sampled using an evolutionary algorithm. We extend this evolutionary procedure to take into account interactions between cells. We obtain a variety of simple bistable networks that we classify into major subtypes. Some have long been proposed in the context of lateral inhibition through the Notch-Delta pathway, some have been more recently considered and others appear to be new and based on mechanisms not previously considered. The results highlight the role of posttranscriptional interactions and particularly of protein complexation and sequestration, which can replace cooperativity in transcriptional interactions. Some bistable networks are entirely based on posttranscriptional interactions and the simplest of these is found to lead, upon a single parameter change, to oscillations in the two cells with opposite phases. We provide qualitative explanations as well as mathematical analyses of the dynamical behaviors of various created networks. The results should help to identify and understand genetic structures implicated in cell-cell interactions and differentiation. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Carbon Heavy-ion Radiation Induced Biological effects on Oryza sativa L.

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Zhang, Meng; Sun, Yeqing; Li, Xishan; Gong, Ning; Meng, Qingmei; Liu, Jiawei; Wang, Ting

2016-07-01

Large number of researches on rice after spaceflights indicated that rice was a favorable model organism to study biological effects induced by space radiation. The stimulative effect could often be found on rice seedlings after irradiation by low-dose energetic heavy-ion radiation. Spaceflight also could induce stimulative effect on kinds of seeds. To further understand the mechanism of low-dose radiation biological effects and the dose range, the germinated rice seeds which were irradiated by different doses of carbon heavy-ion (0, 0.02, 0.1, 0.2, 1, 2, 5, 10, 15 and 20Gy, LET=27.3keV/µm) were used as materials to study. By investigating the variation of rice phenotype under different doses, we found that 2Gy radiation dose was a dividing point of the phenotypic variation. Transmission electron microscopy was used to observe the variation of mitochondria, chloroplast, endoplasmic reticulum, ribosome and nucleus in mesophyll cell of rice apical meristem at 24 hours after radiation with different doses. The cells were not apparently physiologically damaged when the dose of radiation was less than 2Gy. The number of chloroplast did not change significantly, but the number of mitochondria was significantly increased, and gathered around in the chloroplast and endoplasmic reticulum; the obvious lesion of chloroplast and mitochondria were found at the mesophyll cells when radiation dose was higher than 2Gy. The mitochondria were swelling and appearing blurred crest. The chloroplast and mitochondrial mutation rate increased significantly (pmitochondrial was an important organelle involved in the antioxidative systems, its dysfunction could result in the increase of reactive oxygen species and lipid peroxidation. We found that the growth stimulation induced by low-dose radiation mainly occurred at three-leaf stage along with the increasing activity of antioxidase system and damages of lipid peroxidation. We also found that the relative expression of genes sdhb and aox1a

Measurement of leaf hydraulic conductance and stomatal conductance and their responses to irradiance and dehydration using the Evaporative Flux Method (EFM).

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Sack, Lawren; Scoffoni, Christine

2012-12-31

Water is a key resource, and the plant water transport system sets limits on maximum growth and drought tolerance. When plants open their stomata to achieve a high stomatal conductance (gs) to capture CO2 for photosynthesis, water is lost by transpiration(1,2). Water evaporating from the airspaces is replaced from cell walls, in turn drawing water from the xylem of leaf veins, in turn drawing from xylem in the stems and roots. As water is pulled through the system, it experiences hydraulic resistance, creating tension throughout the system and a low leaf water potential (Ψ(leaf)). The leaf itself is a critical bottleneck in the whole plant system, accounting for on average 30% of the plant hydraulic resistance(3). Leaf hydraulic conductance (K(leaf) = 1/ leaf hydraulic resistance) is the ratio of the water flow rate to the water potential gradient across the leaf, and summarizes the behavior of a complex system: water moves through the petiole and through several orders of veins, exits into the bundle sheath and passes through or around mesophyll cells before evaporating into the airspace and being transpired from the stomata. K(leaf) is of strong interest as an important physiological trait to compare species, quantifying the effectiveness of the leaf structure and physiology for water transport, and a key variable to investigate for its relationship to variation in structure (e.g., in leaf venation architecture) and its impacts on photosynthetic gas exchange. Further, K(leaf) responds strongly to the internal and external leaf environment(3). K(leaf) can increase dramatically with irradiance apparently due to changes in the expression and activation of aquaporins, the proteins involved in water transport through membranes(4), and K(leaf) declines strongly during drought, due to cavitation and/or collapse of xylem conduits, and/or loss of permeability in the extra-xylem tissues due to mesophyll and bundle sheath cell shrinkage or aquaporin deactivation(5

Low Doses of Curcuma longa Modulates Cell Migration and Cell-Cell Adhesion.

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de Campos, Paloma Santos; Matte, Bibiana Franzen; Diel, Leonardo Francisco; Jesus, Luciano Henrique; Bernardi, Lisiane; Alves, Alessandro Menna; Rados, Pantelis Varvaki; Lamers, Marcelo Lazzaron

2017-09-01

Cell invasion and metastasis are involved in clinical failures in cancer treatment, and both events require the acquisition of a migratory behavior by tumor cells. Curcumin is a promising natural product with anti-proliferative activity, but its effects on cell migration are still unclear. We evaluated the effects of curcumin on the proliferation, apoptosis, migration, and cell-cell adhesion of keratinocyte, oral squamous cell carcinoma (OSCC), and fibroblast cell lines, as well as in a xenograft model of OSCC. Curcumin (2 μM) decreased cell proliferation in cell lines with mesenchymal characteristics, while cell death was detected only at 50 μM. We observed that highly migratory cells showed a decrease on migration speed and directionality when treated with 2 or 5 μM of curcumin (50% and 40%, respectively, p curcumin dose dependently decreased cell-cell adhesion, especially on tumor-derived spheroids. Also, in a xenograft model with patient-derived OSCC cells, the administration of curcumin decreased tumor growth and aggressiveness when compared with untreated tumors, indicating the potential antitumor effect in oral cancer. These results suggest that lower doses of curcumin can influence several steps involved in tumorigenesis, including migration properties, suggesting a possible use in cancer therapy. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

Nonimmune cells equipped with T-cell-receptor-like signaling for cancer cell ablation.

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Kojima, Ryosuke; Scheller, Leo; Fussenegger, Martin

2018-01-01

The ability to engineer custom cell-contact-sensing output devices into human nonimmune cells would be useful for extending the applicability of cell-based cancer therapies and for avoiding risks associated with engineered immune cells. Here we have developed a new class of synthetic T-cell receptor-like signal-transduction device that functions efficiently in human nonimmune cells and triggers release of output molecules specifically upon sensing contact with a target cell. This device employs an interleukin signaling cascade, whose OFF/ON switching is controlled by biophysical segregation of a transmembrane signal-inhibitory protein from the sensor cell-target cell interface. We further show that designer nonimmune cells equipped with this device driving expression of a membrane-penetrator/prodrug-activating enzyme construct could specifically kill target cells in the presence of the prodrug, indicating its potential usefulness for target-cell-specific, cell-based enzyme-prodrug cancer therapy. Our study also contributes to the advancement of synthetic biology by extending available design principles to transmit extracellular information to cells.

Proliferating cells in psoriatic dermis are comprised primarily of T cells, endothelial cells, and factor XIIIa+ perivascular dendritic cells

International Nuclear Information System (INIS)

Morganroth, G.S.; Chan, L.S.; Weinstein, G.D.; Voorhees, J.J.; Cooper, K.D.

1991-01-01

Determination of the cell types proliferating in the dermis of patients with psoriasis should identify those cells experiencing activation or responding to growth factors in the psoriatic dermal milieu. Toward that end, sections of formalin-fixed biopsies obtained from 3H-deoxyuridine (3H-dU)-injected skin of eight psoriatic patients were immunostained, followed by autoradiography. Proliferating dermal cells exhibit silver grains from tritium emissions. The identity of the proliferating cells could then be determined by simultaneous visualization with antibodies specific for various cell types. UCHL1+ (CD45RO+) T cells (recall antigen-reactive helper T-cell subset) constituted 36.6 +/- 3.1% (mean +/- SEM, n = 6) of the proliferating dermal cells in involved skin, whereas Leu 18+ (CD45RA+) T cells (recall antigen naive T-cell subsets) comprised only 8.7 +/- 1.5% (n = 6). The Factor XIIIa+ dermal perivascular dendritic cell subset (24.9 +/- 1.5% of proliferating dermal cells, n = 6) and Factor VIII+ endothelial cells represented the two other major proliferating populations in lesional psoriatic dermis. Differentiated tissue macrophages, identified by phase microscopy as melanophages or by immunostaining with antibodies to Leu M1 (CD15) or myeloid histiocyte antigen, comprised less than 5% of the proliferating population in either skin type. In addition to calculating the relative proportions of these cells to each other as percent, we also determined the density of cells, in cells/mm2 of tissue. The density of proliferating cells within these populations was increased in involved versus uninvolved skin: UCHL1+, 9.0 +/- 1.7 cells/mm2 versus 1.8 +/- 0.6 cells/mm2, p less than 0.01; Factor XIIIa+, 6.0 +/- 0.7 cells/mm2 versus 1.5 +/- 0.5 cells/mm2, p less than 0.01; Factor VIII+, 5.5 +/- 1.4 cells/mm2 versus 0.0 cells/mm2, p less than 0.05

Factors affecting palatability of four submerged macrophytes for grass carp Ctenopharyngodon idella.

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Sun, Jian; Wang, Long; Ma, Lin; Min, Fenli; Huang, Tao; Zhang, Yi; Wu, Zhenbin; He, Feng

2017-12-01

Grass carp can weaken the growth and reproductive capacity of submerged macrophytes by consuming valuable tissues, but factors affecting palatability of submerged macrophytes for grass carp rarely are considered. In this study, relative consumption rate of grass carp with regard to submerged macrophytes was in the following order: Hydrilla verticillata > Vallisneria natans > Ceratophyllum demersum > Myriophyllum spicatum. Firmness of macrophytes was in the following order: M. spicatum > C. demersum > H. verticillata = V. natans, whereas shear force was M. spicatum > C. demersum > H. verticillata > V. natans. After crude extracts of M. spicatum were combined with H. verticillata, grass carp fed on fewer macrophyte pellets that contained more plant secondary metabolites (PSMs). This indicated that structure and PSMs affected palatability of macrophytes.PSMs do not contribute to reduction in palatability through inhibition of intestinal proteinases activity, but they can cause a decrease in the abundance of Exiguobacterium, Acinetobacter-yielding proteases, lipases, and cellulose activity, which in turn can weaken the metabolic capacity of grass carp and adversely affect their growth. Thus, the disadvantages to the growth and development of grass carp caused by PSMs may drive grass carp to feed on palatable submerged macrophytes with lower PSMs.

Effects of submerged macrophytes on the abundance and community composition of ammonia-oxidizing prokaryotes in a eutrophic lake.

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Zhao, Da-yong; Luo, Juan; Zeng, Jin; Wang, Meng; Yan, Wen-ming; Huang, Rui; Wu, Qinglong L

2014-01-01

Abundances and community compositions of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in unvegetated sediment and the rhizosphere sediments of three submerged macrophytes (Ceratophyllum demersum, Vallisneria spinulosa, and Potamogeton crispus) were investigated in a large, eutrophic freshwater lake, Lake Taihu. Abundances of archaeal ammonia monooxygenase alpha-subunit (amoA) gene (from 6.56 × 10(6) copies to 1.06 × 10(7) copies per gram of dry sediment) were higher than those of bacterial amoA (from 6.13 × 10(5) to 3.21 × 10(6) copies per gram of dry sediment) in all samples. Submerged macrophytes exhibited no significant effect on the abundance and diversity of archaeal amoA gene. C. demersum and V. spinulosa increased the abundance and diversity of bacterial amoA gene in their rhizosphere sediment. However, the diversity of bacterial amoA gene in the rhizosphere sediments of P. crispus was decreased. The data obtained in this study would be helpful to elucidate the roles of submerged macrophytes involved in the nitrogen cycling of eutrophic lake ecosystems.

Removal of Metal Nanoparticles Colloidal Solutions by Water Plants

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Olkhovych, Olga; Svietlova, Nataliia; Konotop, Yevheniia; Karaushu, Olena; Hrechishkina, Svitlana

2016-11-01

The ability of seven species of aquatic plants ( Elodea canadensis, Najas guadelupensis, Vallisneria spiralis L., Riccia fluitans L., Limnobium laevigatum, Pistia stratiotes L., and Salvinia natans L.) to absorb metal nanoparticles from colloidal solutions was studied. It was established that investigated aquatic plants have a high capacity for removal of metal nanoparticles from aqueous solution (30-100%) which indicates their high phytoremediation potential. Analysis of the water samples content for elements including the mixture of colloidal solutions of metal nanoparticles (Mn, Cu, Zn, Ag + Ag2O) before and after exposure to plants showed no significant differences when using submerged or free-floating hydrophytes so-called pleuston. However, it was found that the presence of submerged hydrophytes in aqueous medium ( E. canadensis, N. guadelupensis, V. spiralis L., and R. fluitans L.) and significant changes in the content of photosynthetic pigments, unlike free-floating hydrophytes ( L. laevigatum, P. stratiotes L., S. natans L.), had occur. Pleuston possesses higher potential for phytoremediation of contaminated water basins polluted by metal nanoparticles. In terms of removal of nanoparticles among studied free-floating hydrophytes, P. stratiotes L. and S. natans L. deserve on special attention.

Effects of leaf age within growth stages of pepper and sorghum plants on leaf thickness, water, chlorophyll, and light reflectance. [in spectral vegetation discrimination

Science.gov (United States)

Gausman, H. W.; Cardenas, R.; Berumen, A.

1974-01-01

Pepper and sorghum plants (characterized by porous and compact leaf mesophylls, respectively) were used to study the influence of leaf age on light reflectance. Measurements were limited to the upper five nodal positions within each growth stage, since upper leaves make up most of the reflectance surfaces remotely sensed. The increase in leaf thickness and water content with increasing leaf age was taken into consideration, since each of these factors affects the reflectance as well as the selection of spectral wavelength intervals for optimum discrimination of vegetation.

Pathological significance and prognostic roles of densities of CD57+ cells, CD68+ cells, and mast cells, and their ratios in clear cell renal cell carcinoma.

Science.gov (United States)

Nakanishi, Hiromi; Miyata, Yasuyoshi; Mochizuki, Yasushi; Yasuda, Takuji; Nakamura, Yuichiro; Araki, Kyohei; Sagara, Yuji; Matsuo, Tomohiro; Ohba, Kojiro; Sakai, Hideki

2018-05-19

The immune system is closely associated with malignant behavior in renal cell carcinoma (RCC). Therefore, understanding the pathological roles of immune cells in tumor stroma is essential to discuss the pathological characteristics of RCC. In this study, the clinical significance of densities of CD57+ cells, CD68+ cells, and mast cells, and their ratios were investigated in patients with clear cell RCC. The densities of CD57+, CD68+, and mast cells were evaluated by immunohistochemical techniques in 179 patients. Proliferation index (PI), apoptotic index (AI), and microvessel density (MVD) were evaluated by using anti-Ki-67, anti-cleaved caspase-3, and anti-CD31 antibodies, respectively. The density of CD57+ cell was negatively correlated with grade, pT stage, and metastasis, although densities of CD68+ cell and mast cell were positively correlated. Ratios of CD68+ cell/CD57+ cell and mast cell/CD57+ cell were significantly correlated with grade, pT stage, and metastasis. Survival analyses showed that the CD68+ cell/CD57+ cell ratio was a significant predictor for cause-specific survival by multi-variate analyses (hazard ratio=1.41, 95% confidential interval=1.03-1.93, P=.031), and was significantly correlated with PI, AI, and MVD (r=.47; P <. 001, r=-.31, P<.001, and r=.40, P<.001, respectively). In conclusion, CD57+ cell, CD68+ cell, and mast cell played important roles in malignancy in clear cell RCC. The CD68+ cell/CD57+ cell ratio was strongly correlated with pathological features and prognosis in these patients because this ratio reflected the status of cancer cell proliferation, apoptosis, and angiogenesis. Copyright © 2018. Published by Elsevier Inc.

Validation of leaf ozone symptoms in natural vegetation using microscopical methods

International Nuclear Information System (INIS)

Vollenweider, P.; Ottiger, M.; Guenthardt-Goerg, M.S

2003-01-01

Integration of markers of oxidative stress, from the subcellular to the leaf and needle level, proved to be a useful tool for the differential diagnosis and validation of ozone injury. - Ozone injury to natural vegetation is being increasingly surveyed throughout the northern hemisphere. There exists a growing list of species showing visible 'ozone-like' symptoms which needs to be validated. This study presents the results from a test survey of ozone injury to forest vegetation in the light exposed sites of five Swiss level II plots, for the new ICP-Forests protocol. With AOT40 from 14 to 28 ppm·h in 2000, ten out of 49 woody plant species displayed typical symptoms, and four showed untypical symptoms. Symptom origin was investigated in nine and validated in seven species, using morphological, histological and cellular markers of oxidative stress and ozone-induced plant response. Independent of taxonomic position, ozone effects were characterized by the induction of oxidative stress in the mesophyll resulting in discrete and light-dependent hypersensitive-like responses and in accelerated cell senescence. The presented combination of cellular and morphological markers allows differential diagnosis of visible ozone injury

RNAi-mediated downregulation of poplar plasma membrane intrinsic proteins (PIPs) changes plasma membrane proteome composition and affects leaf physiology.

Science.gov (United States)

Bi, Zhen; Merl-Pham, Juliane; Uehlein, Norbert; Zimmer, Ina; Mühlhans, Stefanie; Aichler, Michaela; Walch, Axel Karl; Kaldenhoff, Ralf; Palme, Klaus; Schnitzler, Jörg-Peter; Block, Katja

2015-10-14

Plasma membrane intrinsic proteins (PIPs) are one subfamily of aquaporins that mediate the transmembrane transport of water. To reveal their function in poplar, we generated transgenic poplar plants in which the translation of PIP genes was downregulated by RNA interference investigated these plants with a comprehensive leaf plasma membrane proteome and physiome analysis. First, inhibition of PIP synthesis strongly altered the leaf plasma membrane protein composition. Strikingly, several signaling components and transporters involved in the regulation of stomatal movement were differentially regulated in transgenic poplars. Furthermore, hormonal crosstalk related to abscisic acid, auxin and brassinosteroids was altered, in addition to cell wall biosynthesis/cutinization, the organization of cellular structures and membrane trafficking. A physiological analysis confirmed the proteomic results. The leaves had wider opened stomata and higher net CO2 assimilation and transpiration rates as well as greater mesophyll conductance for CO2 (gm) and leaf hydraulic conductance (Kleaf). Based on these results, we conclude that PIP proteins not only play essential roles in whole leaf water and CO2 flux but have important roles in the regulation of stomatal movement. Copyright © 2015. Published by Elsevier B.V.

The role of photorespiration during the evolution of C4 photosynthesis in the genus Flaveria.

Science.gov (United States)

Mallmann, Julia; Heckmann, David; Bräutigam, Andrea; Lercher, Martin J; Weber, Andreas P M; Westhoff, Peter; Gowik, Udo

2014-06-16

C4 photosynthesis represents a most remarkable case of convergent evolution of a complex trait, which includes the reprogramming of the expression patterns of thousands of genes. Anatomical, physiological, and phylogenetic and analyses as well as computational modeling indicate that the establishment of a photorespiratory carbon pump (termed C2 photosynthesis) is a prerequisite for the evolution of C4. However, a mechanistic model explaining the tight connection between the evolution of C4 and C2 photosynthesis is currently lacking. Here we address this question through comparative transcriptomic and biochemical analyses of closely related C3, C3-C4, and C4 species, combined with Flux Balance Analysis constrained through a mechanistic model of carbon fixation. We show that C2 photosynthesis creates a misbalance in nitrogen metabolism between bundle sheath and mesophyll cells. Rebalancing nitrogen metabolism requires anaplerotic reactions that resemble at least parts of a basic C4 cycle. Our findings thus show how C2 photosynthesis represents a pre-adaptation for the C4 system, where the evolution of the C2 system establishes important C4 components as a side effect.

Molecular cloning and expression analysis of turnip (Brassica rapa var. rapa sucrose transporter gene family

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Yuanyuan Liu

2017-06-01

Full Text Available In higher plants, sugars (mainly sucrose are produced by photosynthetically assimilated carbon in mesophyll cells of leaves and translocated to heterotrophic organs to ensure plant growth and development. Sucrose transporters, or sucrose carriers (SUCs, play an important role in the long-distance transportation of sucrose from source organs to sink organs, thereby affecting crop yield and quality. The identification, characterization, and molecular function analysis of sucrose transporter genes have been reported for monocot and dicot plants. However, no relevant study has been reported on sucrose transporter genes in Brassica rapa var. rapa, a cruciferous root crop used mainly as vegetables and fodder. We identified and cloned 12 sucrose transporter genes from turnips, named BrrSUC1.1 to BrrSUC6.2 according to the SUC gene sequences of B. rapa pekinensis. We constructed a phylogenetic tree and analyzed conserved motifs for all 12 sucrose transporter genes identified. Real-time quantitative polymerase chain reaction was conducted to understand the expression levels of SUC genes in different tissues and developmental phases of the turnip. These findings add to our understanding of the genetics and physiology of sugar transport during taproot formation in turnips.

How the climate limits the wood density of angiosperms

Science.gov (United States)

Choi, Jin Woo; Kim, Ho-Young

2017-11-01

Flowering trees have various types of wood structure to perform multiple functions under their environmental conditions. In addition to transporting water from the roots to the canopy and providing mechanical support, the structure should provide resistance to embolism to maintain soil-plant-atmosphere continuum. By investigating existing data of the resistivity to embolism and wood density of 165 angiosperm species, here we show that the climate can limit the intrinsic properties of trees. Trees living in the dry environments require a high wood density to slow down the pressure decrease as it loses water relatively fast by evaporation. However, building too much tissues will result in the decrease of hydraulic conductivity and moisture concentration around mesophyll cells. To rationalize the biologically observed lower bound of the wood density, we construct a mechanical model to predict the wood density as a function of the vulnerability to embolism and the time for the recovery. Also, we build an artificial system using hydrogel microchannels that can test the probability of embolism as a function of conduit distributions. Our theoretical prediction is shown to be consistent with the results obtained from the artificial system and the biological data.

Ferredoxin-dependent and antimycin A-sensitive reduction of cytochrome b-559 by far-red light in maize [Zea mays] thylakoids; participation of a meladiol-reducible cytochrome b-559 in cyclic electron flow

International Nuclear Information System (INIS)

Miyake, C.; Schreiber, U.; Asada, K.

1995-01-01

Thylakoids from mesophyll cells of maize showed a high rate of the ferredoxin (Fd)-dependent and antimycin A (AntiA)-sensitive cyclic electron flow as determined by the quenching of 9-aminoacridine fluorescence which indicates the formation of ΔpH across the membranes. Spectrophotometric survey of the thylakoids showed the reduction of a Cyt having an α-peak at 559 nm [Cyt b-559(Fd)] by far-red light, which depended on Fd and was sensitive to AntiA. Dose dependencies of Fd and AntiA on the photoreduction of Cyt b-559(Fd) were the same as those of the formation of ΔpH. Cyt b-559(Fd) occurred in an oxidized form even in the presence of ascorbate and was reduced by far-red light. In darkness, it was reduced only by menadiol (E m,7 = –10mV). Thus, Cyt b-559(Fd) was distinguished from Cyt b-559 in the PSII complex by its low redox potential. The present results indicate that Cyt b-559(Fd) mediates electron transfer from Fd to plastoquinone during Fd-dependent cyclic electron flow around PSI. (author)

LEAF MICROMOPHOMETRY OF PALICOUREA RIGIDA KUNTH. (RUBIACEAE FROM BRAZILIAN CERRADO AND CAMPO RUPESTRE ENVIRONMENTS

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Manuel Losada Gavilanes

2016-06-01

Full Text Available The objective of this work was to evaluate qualitative and quantitative leaf anatomical traits of Palicourea rigida Kunth. (Rubiaceae species occurring in the Brazilian Cerrado and Campo Rupestre ecosystems. Anatomical analysis was performed in fresh or fixed leaves processed with usual plant microtechnique. Leaves showed uniseriate epidermis in petiole and leaf blade which contains uniseriate nonglandular tricomes (tector type occurring only over the vascular bundles. Likewise, paracytic stomata were found only in abaxial side of the leaf surface. The mesophyll contains uniseriate palisade parenchyma and multiseriate spongy parenchyma (nine layers which showed cells with different morphology and size. Crystal idoblasts of different types were observed in both the petiole and leaf blade. Collateral vascular bundles were found both in the petiole and leaf blade. Leaf venation type was pinnate, campylodromous or brochydodromous. The micromorphometric analysis showed significant differences from plants of different environments for all leaf characteristics and Cerrado plants showed higher means for all evaluated traits. Therefore, the influence of environments may had modulated morphological responses in P. rigida, since no difference was found in the type or distribution of leaf tissues in Cerrado or Campo Rupestre.

[Photosynthetic parameters and physiological indexes of Paris polyphylla var. yunnanensis influenced by arbuscular mycorrhizal fungi].

Science.gov (United States)

Wei, Zheng-xin; Guo, Dong-qin; Li, Hai-feng; Ding, Bo; Zhang, Jie; Zhou, Nong; Yu, Jie

2015-10-01

Through potted inoculation test at room temperature and indoor analysis, the photosynthetic parameters and physiological and biochemical indexes of Paris polyphylla var. yunnanensis were observed after 28 arbuscular mycorrhizal (AM) fungi were injected into the P. polyphylla var. yunnanensis growing in a sterile soil environment. The results showed that AM fungi established a good symbiosis with P. polyphylla var. yunnanensis. The AM fungi influenced the photosynthetic parameters and physiological and biochemical indexes of P. polyphylla var. yunnanensis. And the influences were varied depending on different AM fungi. The application of AM fungi improved photosynthesis intensity of P. polyphylla var. yunnanensis mesophyll cells, the contents of soluble protein and soluble sugar, protective enzyme activity of P. polyphylla var. yunnanensis leaf, which was beneficial to resist the adverse environment and promote the growth of P. polyphylla var. yunnanensis. Otherwise, there was a certain mutual selectivity between P. polyphylla var. yunnanensis and AM fungi. From the comprehensive effect of inoculation, Racocetra coralloidea, Scutellospora calospora, Claroideoglomus claroideum, S. pellucida and Rhizophagus clarus were the most suitable AM fungi to P. polyphylla var. yunnanensis when P. polyphylla var. yunnanensis was planted in the field.

Pollination induces autophagy in petunia petals via ethylene.

Science.gov (United States)

Shibuya, Kenichi; Niki, Tomoko; Ichimura, Kazuo

2013-02-01

Autophagy is one of the main mechanisms of degradation and remobilization of macromolecules, and it appears to play an important role in petal senescence. However, little is known about the regulatory mechanisms of autophagy in petal senescence. Autophagic processes were observed by electron microscopy and monodansylcadaverine staining of senescing petals of petunia (Petunia hybrida); autophagy-related gene 8 (ATG8) homologues were isolated from petunia and the regulation of expression was analysed. Nutrient remobilization was also examined during pollination-induced petal senescence. Active autophagic processes were observed in the mesophyll cells of senescing petunia petals. Pollination induced the expression of PhATG8 homologues and was accompanied by an increase in ethylene production. Ethylene inhibitor treatment in pollinated flowers delayed the induction of PhATG8 homologues, and ethylene treatment rapidly upregulated PhATG8 homologues in petunia petals. Dry weight and nitrogen content were decreased in the petals and increased in the ovaries after pollination in detached flowers. These results indicated that pollination induces autophagy and that ethylene is a key regulator of autophagy in petal senescence of petunia. The data also demonstrated the translocation of nutrients from the petals to the ovaries during pollination-induced petal senescence.

Physiological, Ultrastructural and Proteomic Responses in the Leaf of Maize Seedlings to Polyethylene Glycol-Stimulated Severe Water Deficiency

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Ruixin Shao

2015-09-01

Full Text Available After maize seedlings grown in full-strength Hoagland solution for 20 days were exposed to 20% polyethylene glycol (PEG-stimulated water deficiency for two days, plant height, shoot fresh and dry weights, and pigment contents significantly decreased, whereas malondialdehyde (MDA content greatly increased. Using transmission electron microscopy, we observed that chloroplasts of mesophyll cells in PEG-treated maize seedlings were swollen, with a disintegrating envelope and disrupted grana thylakoid lamellae. Using two-dimensional gel electrophoresis (2-DE method, we were able to identify 22 protein spots with significantly altered abundance in the leaves of treated seedlings in response to water deficiency, 16 of which were successfully identified. These protein species were functionally classified into signal transduction, stress defense, carbohydrate metabolism, protein metabolism, and unknown categories. The change in the abundance of the identified protein species may be closely related to the phenotypic and physiological changes due to PEG-stimulated water deficiency. Most of the identified protein species were putatively located in chloroplasts, indicating that chloroplasts may be prone to damage by PEG stimulated-water deficiency in maize seedlings. Our results help clarify the molecular mechanisms of the responses of higher plants to severe water deficiency.

Cell-extracellular matrix and cell-cell adhesion are linked by syndecan-4

DEFF Research Database (Denmark)

Pakideeri Karat, Sandeep Gopal; Multhaupt, Hinke A B; Pocock, Roger

2017-01-01

Cell-extracellular matrix (ECM) and cell-cell junctions that employ microfilaments are sites of tension. They are important for tissue repair, morphogenetic movements and can be emblematic of matrix contraction in fibrotic disease and the stroma of solid tumors. One cell surface receptor, syndecan...... calcium. While it is known that cell-ECM and cell-cell junctions may be linked, possible roles for syndecans in this process are not understood. Here we show that wild type primary fibroblasts and those lacking syndecan-4 utilize different cadherins in their adherens junctions and that tension is a major...... factor in this differential response. This corresponds to the reduced ability of fibroblasts lacking syndecan-4 to exert tension on the ECM and we now show that this may extend to reduced tension in cell-cell adhesion....

MARCKS-related protein regulates cytoskeletal organization at cell-cell and cell-substrate contacts in epithelial cells.

Science.gov (United States)

Van Itallie, Christina M; Tietgens, Amber Jean; Aponte, Angel; Gucek, Marjan; Cartagena-Rivera, Alexander X; Chadwick, Richard S; Anderson, James M

2018-02-02

Treatment of epithelial cells with interferon-γ and TNF-α (IFN/TNF) results in increased paracellular permeability. To identify relevant proteins mediating barrier disruption, we performed proximity-dependent biotinylation (BioID) of occludin and found that tagging of MARCKS-related protein (MRP; also known as MARCKSL1) increased ∼20-fold following IFN/TNF administration. GFP-MRP was focused at the lateral cell membrane and its overexpression potentiated the physiological response of the tight junction barrier to cytokines. However, deletion of MRP did not abrogate the cytokine responses, suggesting that MRP is not required in the occludin-dependent IFN/TNF response. Instead, our results reveal a key role for MRP in epithelial cells in control of multiple actin-based structures, likely by regulation of integrin signaling. Changes in focal adhesion organization and basal actin stress fibers in MRP-knockout (KO) cells were reminiscent of those seen in FAK-KO cells. In addition, we found alterations in cell-cell interactions in MRP-KO cells associated with increased junctional tension, suggesting that MRP may play a role in focal adhesion-adherens junction cross talk. Together, our results are consistent with a key role for MRP in cytoskeletal organization of cell contacts in epithelial cells. © 2018. Published by The Company of Biologists Ltd.

Stem cells engineering for cell-based therapy.

Science.gov (United States)

Taupin, Philippe

2007-09-01

Stem cells carry the promise to cure a broad range of diseases and injuries, from diabetes, heart and muscular diseases, to neurological diseases, disorders and injuries. Significant progresses have been made in stem cell research over the past decade; the derivation of embryonic stem cells (ESCs) from human tissues, the development of cloning technology by somatic cell nuclear transfer (SCNT) and the confirmation that neurogenesis occurs in the adult mammalian brain and that neural stem cells (NSCs) reside in the adult central nervous system (CNS), including that of humans. Despite these advances, there may be decades before stem cell research will translate into therapy. Stem cell research is also subject to ethical and political debates, controversies and legislation, which slow its progress. Cell engineering has proven successful in bringing genetic research to therapy. In this review, I will review, in two examples, how investigators are applying cell engineering to stem cell biology to circumvent stem cells' ethical and political constraints and bolster stem cell research and therapy.

Dissecting engineered cell types and enhancing cell fate conversion via CellNet

Science.gov (United States)

Morris, Samantha A.; Cahan, Patrick; Li, Hu; Zhao, Anna M.; San Roman, Adrianna K.; Shivdasani, Ramesh A.; Collins, James J.; Daley, George Q.

2014-01-01

SUMMARY Engineering clinically relevant cells in vitro holds promise for regenerative medicine, but most protocols fail to faithfully recapitulate target cell properties. To address this, we developed CellNet, a network biology platform that determines whether engineered cells are equivalent to their target tissues, diagnoses aberrant gene regulatory networks, and prioritizes candidate transcriptional regulators to enhance engineered conversions. Using CellNet, we improved B cell to macrophage conversion, transcriptionally and functionally, by knocking down predicted B cell regulators. Analyzing conversion of fibroblasts to induced hepatocytes (iHeps), CellNet revealed an unexpected intestinal program regulated by the master regulator Cdx2. We observed long-term functional engraftment of mouse colon by iHeps, thereby establishing their broader potential as endoderm progenitors and demonstrating direct conversion of fibroblasts into intestinal epithelium. Our studies illustrate how CellNet can be employed to improve direct conversion and to uncover unappreciated properties of engineered cells. PMID:25126792

cgCorrect: a method to correct for confounding cell-cell variation due to cell growth in single-cell transcriptomics

Science.gov (United States)

Blasi, Thomas; Buettner, Florian; Strasser, Michael K.; Marr, Carsten; Theis, Fabian J.

2017-06-01

Accessing gene expression at a single-cell level has unraveled often large heterogeneity among seemingly homogeneous cells, which remains obscured when using traditional population-based approaches. The computational analysis of single-cell transcriptomics data, however, still imposes unresolved challenges with respect to normalization, visualization and modeling the data. One such issue is differences in cell size, which introduce additional variability into the data and for which appropriate normalization techniques are needed. Otherwise, these differences in cell size may obscure genuine heterogeneities among cell populations and lead to overdispersed steady-state distributions of mRNA transcript numbers. We present cgCorrect, a statistical framework to correct for differences in cell size that are due to cell growth in single-cell transcriptomics data. We derive the probability for the cell-growth-corrected mRNA transcript number given the measured, cell size-dependent mRNA transcript number, based on the assumption that the average number of transcripts in a cell increases proportionally to the cell’s volume during the cell cycle. cgCorrect can be used for both data normalization and to analyze the steady-state distributions used to infer the gene expression mechanism. We demonstrate its applicability on both simulated data and single-cell quantitative real-time polymerase chain reaction (PCR) data from mouse blood stem and progenitor cells (and to quantitative single-cell RNA-sequencing data obtained from mouse embryonic stem cells). We show that correcting for differences in cell size affects the interpretation of the data obtained by typically performed computational analysis.

Single cell time-lapse analysis reveals that podoplanin enhances cell survival and colony formation capacity of squamous cell carcinoma cells.

Science.gov (United States)

Miyashita, Tomoyuki; Higuchi, Youichi; Kojima, Motohiro; Ochiai, Atsushi; Ishii, Genichiro

2017-01-06

Tumor initiating cells (TICs) are characterized by high clonal expansion capacity. We previously reported that podoplanin is a TIC-specific marker for the human squamous cell carcinoma cell line A431. The aim of this study is to explore the molecular mechanism underlying the high clonal expansion potential of podoplanin-positive A431cells using Fucci imaging. Single podoplanin-positive cells created large colonies at a significantly higher frequency than single podoplanin-negative cells, whereas no difference was observed between the two types of cells with respect to cell cycle status. Conversely, the cell death ratio of progenies derived from podoplanin-positive single cell was significantly lower than that of cells derived from podoplanin-negative cells. Single A431 cells, whose podoplanin expression was suppressed by RNA interference, exhibited increased cell death ratios and decreased frequency of large colony forming. Moreover, the frequency of large colony forming decreased significantly when podoplanin-positive single cells was treated with a ROCK (Rho-associated coiled-coil kinase) inhibitor, whereas no difference was observed in single podoplanin-negative cells. Our current study cleared that high clonal expansion capacity of podoplanin-positive TICs populations was the result of reduced cell death by podoplanin-mediated signaling. Therefore, podoplanin activity may be a therapeutic target in the treatment of squamous cell carcinomas.

Modeling cell-in-cell structure into its biological significance

OpenAIRE

He, M-f; Wang, S; Wang, Y; Wang, X-n

2013-01-01

Although cell-in-cell structure was noted 100 years ago, the molecular mechanisms of ?entering' and the destination of cell-in-cell remain largely unclear. It takes place among the same type of cells (homotypic cell-in-cell) or different types of cells (heterotypic cell-in-cell). Cell-in-cell formation affects both effector cells and their host cells in multiple aspects, while cell-in-cell death is under more intensive investigation. Given that cell-in-cell has an important role in maintainin...

Cell volume change through water efflux impacts cell stiffness and stem cell fate

NARCIS (Netherlands)

Guo, Ming; Pegoraro, Adrian F.; Mao, Angelo; Zhou, Enhua H.; Arany, Praveen R.; Han, Yulong; Burnette, Dylan T.; Jensen, Mikkel H.; Kasza, Karen E.; Moore, Jeffrey R.; Mackintosh, Frederick C.; Fredberg, Jeffrey J.; Mooney, David J.; Lippincott-Schwartz, Jennifer; Weitz, David A.

2017-01-01

Cells alter their mechanical properties in response to their local microenvironment; this plays a role in determining cell function and can even influence stem cell fate. Here, we identify a robust and unified relationship between cell stiffness and cell volume. As a cell spreads on a substrate, its

Retinal stem cells and potential cell transplantation treatments

Directory of Open Access Journals (Sweden)

Tai-Chi Lin

2014-11-01

Full Text Available The retina, histologically composed of ten delicate layers, is responsible for light perception and relaying electrochemical signals to the secondary neurons and visual cortex. Retinal disease is one of the leading clinical causes of severe vision loss, including age-related macular degeneration, Stargardt's disease, and retinitis pigmentosa. As a result of the discovery of various somatic stem cells, advances in exploring the identities of embryonic stem cells, and the development of induced pluripotent stem cells, cell transplantation treatment for retinal diseases is currently attracting much attention. The sources of stem cells for retinal regeneration include endogenous retinal stem cells (e.g., neuronal stem cells, Müller cells, and retinal stem cells from the ciliary marginal zone and exogenous stem cells (e.g., bone mesenchymal stem cells, adipose-derived stem cells, embryonic stem cells, and induced pluripotent stem cells. The success of cell transplantation treatment depends mainly on the cell source, the timing of cell harvesting, the protocol of cell induction/transplantation, and the microenvironment of the recipient's retina. This review summarizes the different sources of stem cells for regeneration treatment in retinal diseases and surveys the more recent achievements in animal studies and clinical trials. Future directions and challenges in stem cell transplantation are also discussed.

Hybrid cell adhesive material for instant dielectrophoretic cell trapping and long-term cell function assessment.

Science.gov (United States)

Reyes, Darwin R; Hong, Jennifer S; Elliott, John T; Gaitan, Michael

2011-08-16

Dielectrophoresis (DEP) for cell manipulation has focused, for the most part, on approaches for separation/enrichment of cells of interest. Advancements in cell positioning and immobilization onto substrates for cell culture, either as single cells or as cell aggregates, has benefited from the intensified research efforts in DEP (electrokinetic) manipulation. However, there has yet to be a DEP approach that provides the conditions for cell manipulation while promoting cell function processes such as cell differentiation. Here we present the first demonstration of a system that combines DEP with a hybrid cell adhesive material (hCAM) to allow for cell entrapment and cell function, as demonstrated by cell differentiation into neuronlike cells (NLCs). The hCAM, comprised of polyelectrolytes and fibronectin, was engineered to function as an instantaneous cell adhesive surface after DEP manipulation and to support long-term cell function (cell proliferation, induction, and differentiation). Pluripotent P19 mouse embryonal carcinoma cells flowing within a microchannel were attracted to the DEP electrode surface and remained adhered onto the hCAM coating under a fluid flow field after the DEP forces were removed. Cells remained viable after DEP manipulation for up to 8 d, during which time the P19 cells were induced to differentiate into NLCs. This approach could have further applications in areas such as cell-cell communication, three-dimensional cell aggregates to create cell microenvironments, and cell cocultures.

Cell cycle regulation in human embryonic stem cells: links to adaptation to cell culture.

Science.gov (United States)

Barta, Tomas; Dolezalova, Dasa; Holubcova, Zuzana; Hampl, Ales

2013-03-01

Cell cycle represents not only a tightly orchestrated mechanism of cell replication and cell division but it also plays an important role in regulation of cell fate decision. Particularly in the context of pluripotent stem cells or multipotent progenitor cells, regulation of cell fate decision is of paramount importance. It has been shown that human embryonic stem cells (hESCs) show unique cell cycle characteristics, such as short doubling time due to abbreviated G1 phase; these properties change with the onset of differentiation. This review summarizes the current understanding of cell cycle regulation in hESCs. We discuss cell cycle properties as well as regulatory machinery governing cell cycle progression of undifferentiated hESCs. Additionally, we provide evidence that long-term culture of hESCs is accompanied by changes in cell cycle properties as well as configuration of several cell cycle regulatory molecules.

Cell patterning without chemical surface modification: Cell cell interactions between printed bovine aortic endothelial cells (BAEC) on a homogeneous cell-adherent hydrogel

Science.gov (United States)

Chen, C. Y.; Barron, J. A.; Ringeisen, B. R.

2006-10-01

Cell printing offers the unique ability to directly deposit one or multiple cell types directly onto a surface without the need to chemically pre-treat the surface with lithographic methods. We utilize biological laser printing (BioLP ™) to form patterns of bovine aortic endothelial cells (BAECs) onto a homogeneous cell adherent hydrogel surface. These normal cells are shown to retain near-100% viability post-printing. In order to determine whether BAECs encountered shear and/or heat stress during printing, immunocytochemical staining experiments were performed to detect potential expression of heat shock proteins (HSP) by the deposited cells. Printed BAECs expressed HSP at levels similar to negative control cells, indicating that the BioLP process does not expose cells to damaging levels of stress. However, HSP expression was slightly higher at the highest laser energy studied, suggesting more stress was present under these extreme conditions. Printed BAECs also showed preferential asymmetric growth and migration towards each other and away from the originally printed pattern, demonstrating a retained ability for the cells to communicate post-printing.

Llgl1 Connects Cell Polarity with Cell-Cell Adhesion in Embryonic Neural Stem Cells.

Science.gov (United States)

Jossin, Yves; Lee, Minhui; Klezovitch, Olga; Kon, Elif; Cossard, Alexia; Lien, Wen-Hui; Fernandez, Tania E; Cooper, Jonathan A; Vasioukhin, Valera

2017-06-05

Malformations of the cerebral cortex (MCCs) are devastating developmental disorders. We report here that mice with embryonic neural stem-cell-specific deletion of Llgl1 (Nestin-Cre/Llgl1 fl/fl ), a mammalian ortholog of the Drosophila cell polarity gene lgl, exhibit MCCs resembling severe periventricular heterotopia (PH). Immunohistochemical analyses and live cortical imaging of PH formation revealed that disruption of apical junctional complexes (AJCs) was responsible for PH in Nestin-Cre/Llgl1 fl/fl brains. While it is well known that cell polarity proteins govern the formation of AJCs, the exact mechanisms remain unclear. We show that LLGL1 directly binds to and promotes internalization of N-cadherin, and N-cadherin/LLGL1 interaction is inhibited by atypical protein kinase C-mediated phosphorylation of LLGL1, restricting the accumulation of AJCs to the basolateral-apical boundary. Disruption of the N-cadherin-LLGL1 interaction during cortical development in vivo is sufficient for PH. These findings reveal a mechanism responsible for the physical and functional connection between cell polarity and cell-cell adhesion machineries in mammalian cells. Copyright © 2017 Elsevier Inc. All rights reserved.

Snail modulates cell metabolism in MDCK cells

Energy Technology Data Exchange (ETDEWEB)

Haraguchi, Misako, E-mail: haraguci@m3.kufm.kagoshima-u.ac.jp [Department of Biochemistry and Molecular Biology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Indo, Hiroko P. [Department of Maxillofacial Radiology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Iwasaki, Yasumasa [Health Care Center, Kochi University, Kochi 780-8520 (Japan); Iwashita, Yoichiro [Department of Maxillofacial Radiology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Fukushige, Tomoko [Department of Dermatology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Majima, Hideyuki J. [Department of Maxillofacial Radiology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Izumo, Kimiko; Horiuchi, Masahisa [Department of Environmental Medicine, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Kanekura, Takuro [Department of Dermatology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Furukawa, Tatsuhiko [Department of Molecular Oncology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan); Ozawa, Masayuki [Department of Biochemistry and Molecular Biology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima 890-8544 (Japan)

2013-03-22

Highlights: ► MDCK/snail cells were more sensitive to glucose deprivation than MDCK/neo cells. ► MDCK/snail cells had decreased oxidative phosphorylation, O{sub 2} consumption and ATP content. ► TCA cycle enzyme activity, but not expression, was lower in MDCK/snail cells. ► MDCK/snail cells showed reduced PDH activity and increased PDK1 expression. ► MDCK/snail cells showed reduced expression of GLS2 and ACLY. -- Abstract: Snail, a repressor of E-cadherin gene transcription, induces epithelial-to-mesenchymal transition and is involved in tumor progression. Snail also mediates resistance to cell death induced by serum depletion. By contrast, we observed that snail-expressing MDCK (MDCK/snail) cells undergo cell death at a higher rate than control (MDCK/neo) cells in low-glucose medium. Therefore, we investigated whether snail expression influences cell metabolism in MDCK cells. Although gylcolysis was not affected in MDCK/snail cells, they did exhibit reduced pyruvate dehydrogenase (PDH) activity, which controls pyruvate entry into the tricarboxylic acid (TCA) cycle. Indeed, the activity of multiple enzymes involved in the TCA cycle was decreased in MDCK/snail cells, including that of mitochondrial NADP{sup +}-dependent isocitrate dehydrogenase (IDH2), succinate dehydrogenase (SDH), and electron transport Complex II and Complex IV. Consequently, lower ATP content, lower oxygen consumption and increased survival under hypoxic conditions was also observed in MDCK/snail cells compared to MDCK/neo cells. In addition, the expression and promoter activity of pyruvate dehydrogenase kinase 1 (PDK1), which phosphorylates and inhibits the activity of PDH, was increased in MDCK/snail cells, while expression levels of glutaminase 2 (GLS2) and ATP-citrate lyase (ACLY), which are involved in glutaminolysis and fatty acid synthesis, were decreased in MDCK/snail cells. These results suggest that snail modulates cell metabolism by altering the expression and activity of

nduced pluripotent stem cells and cell therapy

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Banu İskender

2013-12-01

Full Text Available Human embryonic stem cells are derived from the inner cell mass of a blastocyst-stage embryo. They hold a huge promise for cell therapy with their self-renewing ability and pluripotency, which is known as the potential to differentiate into all cell types originating from three embryonic germ layers. However, their unique pluripotent feature could not be utilised for therapeutic purposes due to the ethical and legal problems during derivation. Recently, it was shown that the cells from adult tissues could be reverted into embryonic state, thereby restoring their pluripotent feature. This has strenghtened the possiblity of directed differentition of the reprogrammed somatic cells into the desired cell types in vitro and their use in regenerative medicine. Although these cells were termed as induced pluripotent cells, the mechanism of pluripotency has yet to be understood. Still, induced pluripotent stem cell technology is considered to be significant by proposing novel approaches in disease modelling, drug screening and cell therapy. Besides their self-renewing ability and their potential to differentiate into all cell types in a human body, they arouse a great interest in scientific world by being far from the ethical concerns regarding their embryonic counterparts and their unique feature of being patient-specific in prospective cell therapies. In this review, induced pluripotent stem cell technology and its role in cell-based therapies from past to present will be discussed. J Clin Exp Invest 2013; 4 (4: 550-561

Cell-mediated mutagenesis and cell transformation of mammalian cells by chemical carcinogens

International Nuclear Information System (INIS)

Huberman, E.; Langenbach, R.

1977-01-01

We have developed a cell-mediated mutagenesis assay in which cells with the appropriate markers for mutagenesis are co-cultivated with either lethally irradiated rodent embryonic cells that can metabolize carcinogenic hydrocarbons or with primary rat liver cells that can metabolize chemicals carcinogenic to the liver. During co-cultivation, the reactive metabolites of the procarcinogen appear to be transmitted to the mutable cells and induce mutations in them. Assays of this type make it possible to demonstrate a relationship between carcinogenic potency of the chemicals and their ability to induce mutations in mammalian cells. In addition, by simultaneously comparing the frequencies of transformation and mutation induced in normal diploid hamster cells by benzo(a)pyrene (BP) and one of its metabolites, it is possible to estimate the genetic target size for cell transformation in vitro

Gastric stem cells and gastric cancer stem cells

OpenAIRE

Han, Myoung-Eun; Oh, Sae-Ock

2013-01-01

The gastric epithelium is continuously regenerated by gastric stem cells, which give rise to various kinds of daughter cells, including parietal cells, chief cells, surface mucous cells, mucous neck cells, and enteroendocrine cells. The self-renewal and differentiation of gastric stem cells need delicate regulation to maintain the normal physiology of the stomach. Recently, it was hypothesized that cancer stem cells drive the cancer growth and metastasis. In contrast to conventional clonal ev...

Hybrid clone cells derived from human breast epithelial cells and human breast cancer cells exhibit properties of cancer stem/initiating cells.

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Gauck, Daria; Keil, Silvia; Niggemann, Bernd; Zänker, Kurt S; Dittmar, Thomas

2017-08-02

The biological phenomenon of cell fusion has been associated with cancer progression since it was determined that normal cell × tumor cell fusion-derived hybrid cells could exhibit novel properties, such as enhanced metastatogenic capacity or increased drug resistance, and even as a mechanism that could give rise to cancer stem/initiating cells (CS/ICs). CS/ICs have been proposed as cancer cells that exhibit stem cell properties, including the ability to (re)initiate tumor growth. Five M13HS hybrid clone cells, which originated from spontaneous cell fusion events between M13SV1-EGFP-Neo human breast epithelial cells and HS578T-Hyg human breast cancer cells, and their parental cells were analyzed for expression of stemness and EMT-related marker proteins by Western blot analysis and confocal laser scanning microscopy. The frequency of ALDH1-positive cells was determined by flow cytometry using AldeRed fluorescent dye. Concurrently, the cells' colony forming capabilities as well as the cells' abilities to form mammospheres were investigated. The migratory activity of the cells was analyzed using a 3D collagen matrix migration assay. M13HS hybrid clone cells co-expressed SOX9, SLUG, CK8 and CK14, which were differently expressed in parental cells. A variation in the ALDH1-positive putative stem cell population was observed among the five hybrids ranging from 1.44% (M13HS-7) to 13.68% (M13HS-2). In comparison to the parental cells, all five hybrid clone cells possessed increased but also unique colony formation and mammosphere formation capabilities. M13HS-4 hybrid clone cells exhibited the highest colony formation capacity and second highest mammosphere formation capacity of all hybrids, whereby the mean diameter of the mammospheres was comparable to the parental cells. In contrast, the largest mammospheres originated from the M13HS-2 hybrid clone cells, whereas these cells' mammosphere formation capacity was comparable to the parental breast cancer cells. All M13HS

PINCH1 regulates cell-matrix and cell-cell adhesions, cell polarity and cell survival during the peri-implantation stage

DEFF Research Database (Denmark)

Li, Shaohua; Bordoy, Randi; Stanchi, Fabio

2005-01-01

PINCH1 is composed of 5 LIM domains, binds integrin-linked kinase (ILK) and locates to integrin-mediated adhesion sites. In order to investigate PINCH1 function we generated mice and embryonic stem (ES) cell-derived embryoid bodies (EBs) lacking the PINCH1 gene. Similar to mice lacking beta1...... integrin or Ilk, loss of PINCH1 arrested development at the peri-implantation stage. In contrast to beta1 integrin or Ilk mutants, however, disruption of the PINCH1 gene produced implantation chambers with visible cell clumps even at embryonic day 9.5. In order to define the phenotype leading to the peri...... not observed in beta1 integrin- or ILK-deficient mice or EBs, included abnormal cell-cell adhesion of endoderm and epiblast as well as the presence of apoptotic cells in the endodermal cell layer. Although ILK and PINCH1 were shown to be involved in the phosphorylation of serine-473 of PKB/Akt, immunostaining...

The distribution and utilization of nitrogen in developing zea mays L. seedlings

International Nuclear Information System (INIS)

Watt, M.P.M. de O.C.

1987-01-01

During the first five days of germination, in the presence of NO - 3 - 15 N and N-SERVE, the nitrogen reserves of Zea mays L. caryopses accounted for 75% of the total nitrogen content of the seedlings. By day 7, exogenous nitrate contributed to all the inorganic nitrogen measured in the plant. Although NO - 3 - 15 N (94,2 atom % 15 N) was supplied throughout the germination process, and increasing pool of unlabelled nitrate was detected in both the grain and the seedling during this period. It appears that during germination in maize there is an oxidative pathway from reduced nitrogen in the reserve proteins to nitrate-nitrogen which is then supplied to the developing embryo. The levels of nitrate in the leaf increased towards the sheath, whereas other forms of nitrogen, nitrate reductase activity and capacity for nitrate accumulation increased in the opposite direction. Studies with mesophyll and bundle sheath protoplasts showed that the mesophyll tissue contributes over 80% of the total leaf content of inorganic nitrogen. Leaf tissue of Zea mays was found to have the capacity to assimilate nitrate in the dark, but at a lower rate than in the light. Oxygen did not restrict the initial rate of nitrate reduction in the dark. The rate of the in vivo nitrate reduction declined during the second hour of dark anaerobiosis, and was only restored upon supply of oxygen. Under dark anaerobic conditions nitrite accumulated and, on transfer to oxygen, the accumulate nitrite was reduced

Photosynthesis and photosynthetic electron flow in the alpine evergreen species Quercus guyavifolia in winter

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Wei Huang

2016-10-01

Full Text Available Alpine evergreen broadleaf tree species must regularly cope with low night temperatures in winter. However, the effects of low night temperatures on photosynthesis in alpine evergreen broadleaf tree species are unclear. We measured the diurnal photosynthetic parameters before and after cold snap for leaves of Quercus guyavifolia growing in its native habitat at 3290 m. On 11 and 12 December 2013 (before cold snap, stomatal and mesophyll conductances (gs and gm, CO2 assimilation rate (An, and total electron flow through PSII (JPSII at daytime were maintained at high levels. The major action of alternative electron flow was to provide extra ATP for primary metabolisms. On 20 December 2013 (after cold snap, the diurnal values of gs, gm, An and JPSII at daytime largely decreased, mainly due to the large decrease in night air temperature. Meanwhile, the ratio of photorespiration and alternative electron flow to JPSII largely increased on 20 December. Furthermore, the high levels of alternative electron flow were accompanied with low rates of extra ATP production. A quantitative limitation analysis reveals that the gm limitation increased on 20 December with decreased night air temperature. Therefore, the night air temperature was an important determinant of stomatal/mesophyll conductance and photosynthesis. When photosynthesis is inhibited following freezing night temperatures, photorespiration and alternative electron flow are important electron sinks, which support the role of photorespiration and alternative electron flow in photoportection for alpine plants under low temperatures.

Leaf ultraviolet optical properties along a latitudinal gradient in the Arctic-Alpine life zone

International Nuclear Information System (INIS)

Robberecht, R.; Caldwell, M.M.; Billings, W.D.

1980-01-01

Leaf epidermal transmittance of terrestrial solar ultraviolet-B radiation (295 to 320 nm) was examined along a latitudinal gradient of solar uv-B radiation. In high uv-B radiation zones, e.g., equatorial and tropical regions, mean epidermal transmittance for the species examined was less than 2%. At higher latitudes, mean epidermal transmittance exceeded 5%. Although this latitudinal solar uv-B gradient represents more than a seven-fold difference in daily integrated uv-B irradiance, the calculated mean effective uv-B irradiance at the mesophyll of low-latitude species is not substantially different from that of species at higher latitudes. Species in high uv-B radiation environments appear to attenuate this radiation more effectively than those in lower irradiance environments. In most cases, absorption of uv-B in the epidermis is the major parameter effecting low transmittance. Reflectance from glabrous leaves is generally less than 10%. In some species, pubescent or glaucous leaf surfaces can reflect more than 40% of the uv-B radiation incident on a horizontal leaf, although such surface characteristics do not necessarily indicate high uv-B reflectance. Under controlled conditions, epidermal transmittance in Pisum sativum L. decreased in response to uv-B irradiation. The modification of epidermal transmittance, resulting in lower uv-B irradiance at the mesophyll, may represent a mechanism of plant acclimation to uv-B radiation. Such acclimation may have occurred in several wildland species of temperate-latitude origin that have invaded high uv-B irradiance equatorial and tropical regions

Penetration of UV-A, UV-B, blue, and red light into leaf tissues of pecan measured by a fiber optic microprobe system

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Qi, Yadong; Bai, Shuju; Vogelmann, Thomas C.; Heisler, Gordon M.

2003-11-01

The depth of light penetration from the adaxial surfaces of the mature leaves of pecan (Carya illinoensis) was measured using a fiber optic microprobe system at four wavelengths: UV-B (310nm), UV-A (360 nm), blue light (430nm), and red light (680nm). The average thickness of the leaf adaxial epidermal layer was 15um and the total leaf thickness was 219um. The patterns of the light attenuation by the leaf tissues exhibited strong wavelength dependence. The leaf adaxial epidermal layer was chiefly responsible for absorbing the UV-A UV-B radiation. About 98% of 310 nm light was steeply attenuated within the first 5 um of the adaxial epidermis; thus, very little UV-B radiation was transmitted to the mesophyll tissues where contain photosynthetically sensitive sites. The adaxial epidermis also attenuated 96% of the UV-A radiation. In contrast, the blue and red light penetrated much deeper and was gradually attenutated by the leaves. The mesophyll tissues attenuated 17% of the blue light and 42% of the red light, which were available for photosynthesis use. Since the epidermal layer absorbed nearly all UV-B light, it acted as an effective filter screening out the harmful radiation and protecting photosynthetically sensitive tissues from the UV-B damage. Therefore, the epidermal function of the UV-B screening effectiveness can be regarded as one of the UV-B protection mechanisms in pecan.

In vitro differentiation of primordial germ cells and oocyte-like cells from stem cells.

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Costa, José J N; Souza, Glaucinete B; Soares, Maria A A; Ribeiro, Regislane P; van den Hurk, Robert; Silva, José R V

2018-02-01

Infertility is the result of failure due to an organic disorder of the reproductive organs, especially their gametes. Recently, much progress has been made on generating germ cells, including oocytes, from various types of stem cells. This review focuses on advances in female germ cell differentiation from different kinds of stem cells, with emphasis on embryonic stem cells, adult stem cells, and induced pluripotent stem cells. The advantages and disadvantages of the derivation of female germ cells from several types of stem cells are also highlighted, as well as the ability of stem cells to generate mature and functional female gametes. This review shows that stem cell therapies have opened new frontiers in medicine, especially in the reproductive area, with the possibility of regenerating fertility.

Cell cycle control by components of cell anchorage

OpenAIRE

Gad, Annica

2005-01-01

Extracellular factors, such as growth factors and cell anchorage to the extracellular matrix, control when and where cells may proliferate. This control is abolished when a normal cell transforms into a tumour cell. The control of cell proliferation by cell anchorage was elusive and less well studied than the control by growth factors. Therefore, we aimed to clarify at what points in the cell cycle and through which molecular mechanisms cell anchorage controls cell cycle pro...

Casticin impairs cell growth and induces cell apoptosis via cell cycle arrest in human oral cancer SCC-4 cells.

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Chou, Guan-Ling; Peng, Shu-Fen; Liao, Ching-Lung; Ho, Heng-Chien; Lu, Kung-Wen; Lien, Jin-Cherng; Fan, Ming-Jen; La, Kuang-Chi; Chung, Jing-Gung

2018-02-01

Casticin, a polymethoxyflavone, present in natural plants, has been shown to have biological activities including anti-cancer activities. Herein, we investigated the anti-oral cancer activity of casticin on SCC-4 cells in vitro. Viable cells, cell cycle distribution, apoptotic cell death, reactive oxygen species (ROS) production, and Ca 2+ production, levels of ΔΨ m and caspase activity were measured by flow cytometric assay. Cell apoptosis associated protein expressions were examined by Western blotting and confocal laser microscopy. Results indicated that casticin induced cell morphological changes, DNA condensation and damage, decreased the total viable cells, induced G 2 /M phase arrest in SCC-4 cells. Casticin promoted ROS and Ca 2+ productions, decreases the levels of ΔΨ m , promoted caspase-3, -8, and -9 activities in SCC-4 cells. Western blotting assay demonstrated that casticin affect protein level associated with G2/M phase arrest and apoptosis. Confocal laser microscopy also confirmed that casticin increased the translocation of AIF and cytochrome c in SCC-4 cells. In conclusion, casticin decreased cell number through G 2 /M phase arrest and the induction of cell apoptosis through caspase- and mitochondria-dependent pathways in SCC-4 cells. © 2017 Wiley Periodicals, Inc.

Skin Stem Cells in Skin Cell Therapy

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Mollapour Sisakht

2015-12-01

Full Text Available Context Preclinical and clinical research has shown that stem cell therapy is a promising therapeutic option for many diseases. This article describes skin stem cells sources and their therapeutic applications. Evidence Acquisition Compared with conventional methods, cell therapy reduces the surgical burden for patients because it is simple and less time-consuming. Skin cell therapy has been developed for variety of diseases. By isolation of the skin stem cell from the niche, in vitro expansion and transplantation of cells offers a surprising healing capacity profile. Results Stem cells located in skin cells have shown interesting properties such as plasticity, transdifferentiation, and specificity. Mesenchymal cells of the dermis, hypodermis, and other sources are currently being investigated to promote regeneration. Conclusions Because skin stem cells are highly accessible from autologous sources and their immunological profile is unique, they are ideal for therapeutic approaches. Optimization of administrative routes requires more investigation own to the lack of a standard protocol.

Cell biology of mesangial cells: the third cell that maintains the glomerular capillary.

Science.gov (United States)

Kurihara, Hidetake; Sakai, Tatsuo

2017-03-01

The renal glomerulus consists of glomerular endothelial cells, podocytes, and mesangial cells, which cooperate with each other for glomerular filtration. We have produced monoclonal antibodies against glomerular cells in order to identify different types of glomerular cells. Among these antibodies, the E30 clone specifically recognizes the Thy1.1 molecule expressed on mesangial cells. An injection of this antibody into rats resulted in mesangial cell-specific injury within 15 min, and induced mesangial proliferative glomerulonephritis in a reproducible manner. We examined the role of mesangial cells in glomerular function using several experimental tools, including an E30-induced nephritis model, mesangial cell culture, and the deletion of specific genes. Herein, we describe the characterization of E30-induced nephritis, formation of the glomerular capillary network, mesangial matrix turnover, and intercellular signaling between glomerular cells. New molecules that are involved in a wide variety of mesangial cell functions are also introduced.

Natural killer cells for immunotherapy – Advantages of cell lines over blood NK cells

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Hans eKlingemann

2016-03-01

Full Text Available Natural killer cells are potent cytotoxic effector cells for cancer therapy and potentially for severe viral infections. However, there are technical challenges to obtain sufficient numbers of functionally active NK cells form a patient’s blood since they represent only 10% of the lymphocytes. Especially, cancer patients are known to have dysfunctional NK cells. The alternative is to obtain cells from a healthy donor, which requires depletion of the allogeneic T-cells. Establishing cell lines from donor blood NK cells have not been successful, in contrast to blood NK cells obtained from patients with a clonal NK cell lymphoma. Those cells can be expanded in culture in the presence of IL-2. However, except for the NK-92 cell line none of the other six known cell lines has consistent and reproducibly high anti-tumor cytotoxicity, nor can they be easily genetically manipulated to recognize specific tumor antigens or to augment monoclonal antibody activity through ADCC. NK-92 is also the only cell line product that has been widely given to patients with advanced cancer with demonstrated efficiency and minimal side effects.

Cell sheet technology and cell patterning for biofabrication

Energy Technology Data Exchange (ETDEWEB)

Hannachi, Imen Elloumi; Yamato, Masayuki; Okano, Teruo [Institute of Advanced Biomedical Engineering and Science, Tokyo Women' s Medical University, 8-1 Kawada-cho, Shinjuku, Tokyo (Japan)

2009-06-01

We have developed cell sheet technology as a modern method for the fabrication of functional tissue-like and organ-like structures. This technology allows for a sheet of interconnected cells and cells in full contact with their natural extracellular environment to be obtained. A cell sheet can be patterned and composed according to more than one cell type. The key technology of cell sheet engineering is that a fabricated cell sheet can be harvested and transplanted utilizing temperature-responsive surfaces. In this review, we summarize different aspects of cell sheet engineering and provide a survey of the application of cell sheets as a suitable material for biofabrication and clinics. Moreover, since cell micropatterning is a key tool for cell sheet engineering, in this review we focus on the introduction of our approaches to cell micropatterning and cell co-culture to the principles of automation and how they can be subjected to easy robotics programming. Finally, efforts towards making cell sheet technology suitable for biofabrication and robotic biofabrication are also summarized. (topical review)

Free-zone electrophoresis of animal cells. 1: Experiments on cell-cell interactions

Science.gov (United States)

Todd, P. W.; Hjerten, S.

1985-01-01

The electrophoretically migrating zones wasa monitored. The absence of fluid flows in the direction of migration permits direct measurement of electrophoretic velocities of any material. Sedimentation is orthogonal to electrokinetic motion and the effects of particle-particle interaction on electrophoretic mobility is studied by free zone electrophoresis. Fixed erythrocytes at high concentrations, mixtures of fixed erythrocytes from different animal species, and mixtures of cultured human cells were studied in low ionic strength buffers. The electrophoretic velocity of fixed erythrocytes was not altered by increasing cell concentration or by the mixing of erythrocytes from different species. When zones containing cultured human glial cells and neuroblastoma cells are permitted to interact during electrophoresis, altered migration patterns occur. It is found that cell-cell interactions depends upon cell type.

Characterization of a Merkel Cell Polyomavirus-Positive Merkel Cell Carcinoma Cell Line CVG-1.

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Velásquez, Celestino; Amako, Yutaka; Harold, Alexis; Toptan, Tuna; Chang, Yuan; Shuda, Masahiro

2018-01-01

Merkel cell polyomavirus (MCV) plays a causal role in ∼80% of Merkel cell carcinomas (MCC). MCV is clonally integrated into the MCC tumor genome, which results in persistent expression of large T (LT) and small T (sT) antigen oncoproteins encoded by the early locus. In MCV-positive MCC tumors, LT is truncated by premature stop codons or deletions that lead to loss of the C-terminal origin binding (OBD) and helicase domains important for replication. The N-terminal Rb binding domain remains intact. MCV-positive cell lines derived from MCC explants have been valuable tools to study the molecular mechanism of MCV-induced Merkel cell carcinogenesis. Although all cell lines have integrated MCV and express truncated LT antigens, the molecular sizes of the LT proteins differ between cell lines. The copy number of integrated viral genome also varies across cell lines, leading to significantly different levels of viral protein expression. Nevertheless, these cell lines share phenotypic similarities in cell morphology, growth characteristics, and neuroendocrine marker expression. Several low-passage MCV-positive MCC cell lines have been established since the identification of MCV. We describe a new MCV-positive MCV cell line, CVG-1, with features distinct from previously reported cell lines. CVG-1 tumor cells grow in more discohesive clusters in loose round cell suspension, and individual cells show dramatic size heterogeneity. It is the first cell line to encode an MCV sT polymorphism resulting in a unique leucine (L) to proline (P) substitution mutation at amino acid 144. CVG-1 possesses a LT truncation pattern near identical to that of MKL-1 cells differing by the last two C-terminal amino acids and also shows an LT protein expression level similar to MKL-1. Viral T antigen knockdown reveals that, like other MCV-positive MCC cell lines, CVG-1 requires T antigen expression for cell proliferation.

c-Myc-Dependent Cell Competition in Human Cancer Cells.

Science.gov (United States)

Patel, Manish S; Shah, Heta S; Shrivastava, Neeta

2017-07-01

Cell Competition is an interaction between cells for existence in heterogeneous cell populations of multicellular organisms. This phenomenon is involved in initiation and progression of cancer where heterogeneous cell populations compete directly or indirectly for the survival of the fittest based on differential gene expression. In Drosophila, cells having lower dMyc expression are eliminated by cell competition through apoptosis when present in the milieu of cells having higher dMyc expression. Thus, we designed a study to develop c-Myc (human homolog) dependent in vitro cell competition model of human cancer cells. Cells with higher c-Myc were transfected with c-myc shRNA to prepare cells with lower c-Myc and then co-cultured with the same type of cells having a higher c-Myc in equal ratio. Cells with lower c-Myc showed a significant decrease in numbers when compared with higher c-Myc cells, suggesting "loser" and "winner" status of cells, respectively. During microscopy, engulfment of loser cells by winner cells was observed with higher expression of JNK in loser cells. Furthermore, elimination of loser cells was prevented significantly, when co-cultured cells were treated with the JNK (apoptosis) inhibitor. Above results indicate elimination of loser cells in the presence of winner cells by c-Myc-dependent mechanisms of cell competition in human cancer cells. This could be an important mechanism in human tumors where normal cells are eliminated by c-Myc-overexpressed tumor cells. J. Cell. Biochem. 118: 1782-1791, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

β-Cell regeneration through the transdifferentiation of pancreatic cells: Pancreatic progenitor cells in the pancreas.

Science.gov (United States)

Kim, Hyo-Sup; Lee, Moon-Kyu

2016-05-01

Pancreatic progenitor cell research has been in the spotlight, as these cells have the potential to replace pancreatic β-cells for the treatment of type 1 and 2 diabetic patients with the absence or reduction of pancreatic β-cells. During the past few decades, the successful treatment of diabetes through transplantation of the whole pancreas or isolated islets has nearly been achieved. However, novel sources of pancreatic islets or insulin-producing cells are required to provide sufficient amounts of donor tissues. To overcome this limitation, the use of pancreatic progenitor cells is gaining more attention. In particular, pancreatic exocrine cells, such as duct epithelial cells and acinar cells, are attractive candidates for β-cell regeneration because of their differentiation potential and pancreatic lineage characteristics. It has been assumed that β-cell neogenesis from pancreatic progenitor cells could occur in pancreatic ducts in the postnatal stage. Several studies have shown that insulin-producing cells can arise in the duct tissue of the adult pancreas. Acinar cells also might have the potential to differentiate into insulin-producing cells. The present review summarizes recent progress in research on the transdifferentiation of pancreatic exocrine cells into insulin-producing cells, especially duct and acinar cells.

Intrinsic Plasma Cell Differentiation Defects in B Cell Expansion with NF-κB and T Cell Anergy Patient B Cells

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Swadhinya Arjunaraja

2017-08-01

Full Text Available B cell Expansion with NF-κB and T cell Anergy (BENTA disease is a novel B cell lymphoproliferative disorder caused by germline, gain-of-function mutations in the lymphocyte scaffolding protein CARD11, which drives constitutive NF-κB signaling. Despite dramatic polyclonal expansion of naive and immature B cells, BENTA patients also present with signs of primary immunodeficiency, including markedly reduced percentages of class-switched/memory B cells and poor humoral responses to certain vaccines. Using purified naive B cells from our BENTA patient cohort, here we show that BENTA B cells exhibit intrinsic defects in B cell differentiation. Despite a profound in vitro survival advantage relative to normal donor B cells, BENTA patient B cells were severely impaired in their ability to differentiate into short-lived IgDloCD38hi plasmablasts or CD138+ long-lived plasma cells in response to various stimuli. These defects corresponded with diminished IgG antibody production and correlated with poor induction of specific genes required for plasma cell commitment. These findings provide important mechanistic clues that help explain both B cell lymphocytosis and humoral immunodeficiency in BENTA disease.

Activated Allogeneic NK Cells Preferentially Kill Poor Prognosis B-Cell Chronic Lymphocytic Leukemia Cells.

Science.gov (United States)

Sánchez-Martínez, Diego; Lanuza, Pilar M; Gómez, Natalia; Muntasell, Aura; Cisneros, Elisa; Moraru, Manuela; Azaceta, Gemma; Anel, Alberto; Martínez-Lostao, Luis; Villalba, Martin; Palomera, Luis; Vilches, Carlos; García Marco, José A; Pardo, Julián

2016-01-01

Mutational status of TP53 together with expression of wild-type (wt) IGHV represents the most widely accepted biomarkers, establishing a very poor prognosis in B-cell chronic lymphocytic leukemia (B-CLL) patients. Adoptive cell therapy using allogeneic HLA-mismatched Natural killer (NK) cells has emerged as an effective and safe alternative in the treatment of acute myeloid and lymphoid leukemias that do not respond to traditional therapies. We have described that allogeneic activated NK cells eliminate hematological cancer cell lines with multidrug resistance acquired by mutations in the apoptotic machinery. This effect depends on the activation protocol, being B-lymphoblastoid cell lines (LCLs) the most effective stimulus to activate NK cells. Here, we have further analyzed the molecular determinants involved in allogeneic NK cell recognition and elimination of B-CLL cells, including the expression of ligands of the main NK cell-activating receptors (NKG2D and NCRs) and HLA mismatch. We present preliminary data suggesting that B-CLL susceptibility significantly correlates with HLA mismatch between NK cell donor and B-CLL patient. Moreover, we show that the sensitivity of B-CLL cells to NK cells depends on the prognosis based on TP53 and IGHV mutational status. Cells from patients with worse prognosis (mutated TP53 and wt IGHV ) are the most susceptible to activated NK cells. Hence, B-CLL prognosis may predict the efficacy of allogenic activated NK cells, and, thus, NK cell transfer represents a good alternative to treat poor prognosis B-CLL patients who present a very short life expectancy due to lack of effective treatments.

Lung cells support osteosarcoma cell migration and survival.

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Yu, Shibing; Fourman, Mitchell Stephen; Mahjoub, Adel; Mandell, Jonathan Brendan; Crasto, Jared Anthony; Greco, Nicholas Giuseppe; Weiss, Kurt Richard

2017-01-25

Osteosarcoma (OS) is the most common primary bone tumor, with a propensity to metastasize to the lungs. Five-year survival for metastatic OS is below 30%, and has not improved for several decades despite the introduction of multi-agent chemotherapy. Understanding OS cell migration to the lungs requires an evaluation of the lung microenvironment. Here we utilized an in vitro lung cell and OS cell co-culture model to explore the interactions between OS and lung cells, hypothesizing that lung cells would promote OS cell migration and survival. The impact of a novel anti-OS chemotherapy on OS migration and survival in the lung microenvironment was also examined. Three human OS cell lines (SJSA-1, Saos-2, U-2) and two human lung cell lines (HULEC-5a, MRC-5) were cultured according to American Type Culture Collection recommendations. Human lung cell lines were cultured in growth medium for 72 h to create conditioned media. OS proliferation was evaluated in lung co-culture and conditioned media microenvironment, with a murine fibroblast cell line (NIH-3 T3) in fresh growth medium as controls. Migration and invasion were measured using a real-time cell analysis system. Real-time PCR was utilized to probe for Aldehyde Dehydrogenase (ALDH1) expression. Osteosarcoma cells were also transduced with a lentivirus encoding for GFP to permit morphologic analysis with fluorescence microscopy. The anti-OS efficacy of Disulfiram, an ALDH-inhibitor previously shown to inhibit OS cell proliferation and metastasis in vitro, was evaluated in each microenvironment. Lung-cell conditioned medium promoted osteosarcoma cell migration, with a significantly higher attractive effect on all three osteosarcoma cell lines compared to basic growth medium, 10% serum containing medium, and NIH-3 T3 conditioned medium (p cell conditioned medium induced cell morphologic changes, as demonstrated with GFP-labeled cells. OS cells cultured in lung cell conditioned medium had increased alkaline

Human regulatory B cells control the TFH cell response.

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Achour, Achouak; Simon, Quentin; Mohr, Audrey; Séité, Jean-François; Youinou, Pierre; Bendaoud, Boutahar; Ghedira, Ibtissem; Pers, Jacques-Olivier; Jamin, Christophe

2017-07-01

Follicular helper T (T FH ) cells support terminal B-cell differentiation. Human regulatory B (Breg) cells modulate cellular responses, but their control of T FH cell-dependent humoral immune responses is unknown. We sought to assess the role of Breg cells on T FH cell development and function. Human T cells were polyclonally stimulated in the presence of IL-12 and IL-21 to generate T FH cells. They were cocultured with B cells to induce their terminal differentiation. Breg cells were included in these cultures, and their effects were evaluated by using flow cytometry and ELISA. B-cell lymphoma 6, IL-21, inducible costimulator, CXCR5, and programmed cell death protein 1 (PD-1) expressions increased on stimulated human T cells, characterizing T FH cell maturation. In cocultures they differentiated B cells into CD138 + plasma and IgD - CD27 + memory cells and triggered immunoglobulin secretions. Breg cells obtained by Toll-like receptor 9 and CD40 activation of B cells prevented T FH cell development. Added to T FH cell and B-cell cocultures, they inhibited B-cell differentiation, impeded immunoglobulin secretions, and expanded Foxp3 + CXCR5 + PD-1 + follicular regulatory T cells. Breg cells modulated IL-21 receptor expressions on T FH cells and B cells, and their suppressive activities involved CD40, CD80, CD86, and intercellular adhesion molecule interactions and required production of IL-10 and TGF-β. Human Breg cells control T FH cell maturation, expand follicular regulatory T cells, and inhibit the T FH cell-mediated antibody secretion. These novel observations demonstrate a role for the Breg cell in germinal center reactions and suggest that deficient activities might impair the T FH cell-dependent control of humoral immunity and might lead to the development of aberrant autoimmune responses. Copyright © 2016 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Diffusion and bulk flow in phloem loading: A theoretical analysis of the polymer trap mechanism for sugar transport in plants

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Dölger, Julia; Rademaker, Hanna; Liesche, Johannes; Schulz, Alexander; Bohr, Tomas

2014-10-01

Plants create sugar in the mesophyll cells of their leaves by photosynthesis. This sugar, mostly sucrose, has to be loaded via the bundle sheath into the phloem vascular system (the sieve elements), where it is distributed to growing parts of the plant. We analyze the feasibility of a particular loading mechanism, active symplasmic loading, also called the polymer trap mechanism, where sucrose is transformed into heavier sugars, such as raffinose and stachyose, in the intermediary-type companion cells bordering the sieve elements in the minor veins of the phloem. Keeping the heavier sugars from diffusing back requires that the plasmodesmata connecting the bundle sheath with the intermediary cell act as extremely precise filters, which are able to distinguish between molecules that differ by less than 20% in size. In our modeling, we take into account the coupled water and sugar movement across the relevant interfaces, without explicitly considering the chemical reactions transforming the sucrose into the heavier sugars. Based on the available data for plasmodesmata geometry, sugar concentrations, and flux rates, we conclude that this mechanism can in principle function, but that it requires pores of molecular sizes. Comparing with the somewhat uncertain experimental values for sugar export rates, we expect the pores to be only 5%-10% larger than the hydraulic radius of the sucrose molecules. We find that the water flow through the plasmodesmata, which has not been quantified before, contributes only 10%-20% to the sucrose flux into the intermediary cells, while the main part is transported by diffusion. On the other hand, the subsequent sugar translocation into the sieve elements would very likely be carried predominantly by bulk water flow through the plasmodesmata. Thus, in contrast to apoplasmic loaders, all the necessary water for phloem translocation would be supplied in this way with no need for additional water uptake across the plasma membranes of the

The effect and fate of water-soluble carbon nanodots in maize (Zea mays L.).

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Chen, Jing; Dou, Runzhi; Yang, Zhongzhou; Wang, Xiaoping; Mao, Chuanbin; Gao, Xiang; Wang, Li

2016-08-01

In this study, the toxicity of water-soluble carbon nanodots (C-dots) to maize (Zea mays L.) and their uptake and transport in plants were investigated. After exposed in sand matrix amended with 0-2000 mg/L C-dots for 4 weeks, we found that the phytotoxicity of C-dots was concentration-dependent. C-dots at 250 and 500 mg/L showed no toxicity to maize. However, 1000 and 2000 mg/L C-dots significantly reduced the fresh weight of root by 57% and 68%, and decreased the shoot fresh weight by 38% and 72%, respectively. Moreover, in maize roots, the exposure of C-dots at 2000 mg/L significantly increased the H2O2 content and lipid peroxidation (6.5 and 1.65 times higher, respectively), as well as, the antioxidant enzymes activities, up to 2, 1.5, 1.9 and 1.9 times higher for catalase, ascorbate peroxidase, guaiacol peroxidase and superoxide dismutase, respectively. On the other hand, C-dots were observed in detached root-cap cells, cortex and vascular bundle of roots and mesophyll cells of leaves through fluorescence microscopy analysis, suggesting that C-dots were absorbed and translocated systemically in maize. Remarkably, a certain amount of C-dots were excreted out from leaf blade. To our knowledge, this is the first study combined phenotypic observation with physiologic responses and bioaccumulation and translocation analysis of C-dots to investigate their effect and fate in maize.

Identification of heat-sensitive QTL derived from common wild rice (Oryza rufipogon Griff.).

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Lei, Dongyang; Tan, Lubin; Liu, Fengxia; Chen, Liyun; Sun, Chuanqing

2013-03-01

Understanding the responses of rice plants to heat-stress is a challenging, yet crucial, endeavor. A set of introgression lines was previously developed using an advanced backcrossing strategy that involved the elite indica cultivar Teqing as the recipient and an accession of common wild rice (Oryza rufipongon Griff.) as the donor. In this study, we evaluated the responses of 90 of these previously developed introgression lines to heat stress. Five quantitative trait loci (QTLs) related to heat response were detected. The phenotypic variances explained by these QTLs ranged from 6.83% to 14.63%, and O. rufipogon-derived alleles at one locus reduced sensitivity to heat. A heat-sensitive introgression line, YIL106, was identified and characterized. Genotypic analysis demonstrated that YIL106 contained four introgressed segments derived from O. rufipongon and two QTLs (qHTS1-1 and qHTS3) related to heat response. Physiological tests, including measurements of chlorophyll content, electrolyte leakage, malondialdehyde content, and soluble sugar content, were consistent with the heat sensitivity observed in YIL106. Ultrastructural analysis of YIL106 mesophyll cells showed that they were severely damaged following heat stress. This suggests that modification of the cell membrane system is a primary response to heat stress in plants. Identification and characterization of the heat-sensitive line YIL106 may facilitate the isolation of genes associated with the response of rice plants to heat stress. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Hydrocarbons and energy from plants: Final report, 1984-1987

Energy Technology Data Exchange (ETDEWEB)

Calvin, M.; Otvos, J.; Taylor, S.E.; Nemethy, E.K.; Skrukrud, C.L.; Hawkins, D.R.; Lago, R.

1988-08-01

Plant hydrocarbon (isoprenoid) production was investigated as an alternative source to fossil fuels. Because of their high triterpenoid (hydrocarbon) content of 4--8%, Euphorbia lathyris plants were used as a model system for this study. The structure of the E. lathyris triterpenoids was determined, and triterpenoid biosynthesis studied to better understand the metabolic regulation of isoprenoid production. Triterpenoid biosynthesis occurs in two distinct tissue types in E. lathyris plants: in the latex of the laticifer cells; and in the mesophyll cells of the leaf and stem. The latex has been fractionated by centrifugation, and it has been determined that the later steps of isoprenoid biosynthesis, the conversion of mevalonic acid to the triterpenes, are compartmentized within a vacuole. Also identified was the conversion of hydroxymethyl glutaryl-CoA to mevalonic acid, catalyzed by the enzyme Hydroxymethyl glutaryl-CoA Reductase, as a key rate limiting step in isoprenoid biosynthesis. At least two isozymes of this enzyme, one in the latex and another in the leaf plastids, have been identified. Environmental stress has been applied to plants to study changes in carbon allocation. Salinity stress caused a large decrease in growth, smaller decreases in photosynthesis, resulting in a larger allocation of carbon to both hydrocarbon and sugar production. An increase in Hydroxymethyl glutaryl-CoA Reductase activity was also observed when isoprenoid production increased. Other species where also screened for the production of hydrogen rich products such as isoprenoids and glycerides, and their hydrocarbon composition was determined.

Electrical signalling along the phloem and its physiological responses in the maize leaf

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Joerg eFromm

2013-07-01

Full Text Available To elucidate the role of electrical signalling in the phloem of maize the tips of attached leaves were stimulated by chilling and wounding. Two different signals were detected in the phloem at the middle of the leaf using the aphid stylet technique: (i action potentials (AP arose in the phloem after chilling; and (ii variation potentials (VP were evoked after wounding the leaf tip. Combined electric potential and gas exchange measurements showed that while the wound-induced VP moved rapidly towards the middle of the leaf to induce a reduction in both the net-CO2 uptake rate and the stomatal conductance, there was no response in the gas exchange to the cold-induced AP. To determine if electrical signalling had any impact on assimilate transport the middle of the leaf was exposed to 14CO2. Autoradiography of labelled assimilates provided evidence that phloem and intercellular transport of assimilates from mesophyll to bundle sheath cells was strongly reduced while the cold-induced AP moved through. In contrast, wound-induced VP did not inhibit assimilate translocation but did reduce the amount of the labelled assimilate in phloem and bundle sheath cells. Biochemical analysis revealed that callose content increased significantly in chilled leaves while starch increased in chilled but decreased in wounded leaves. The results led to the conclusion that different stimulation types incite characteristic phloem-transmitted electrical signals, each with a specific influence on gas exchange and assimilate transport.

Integrating transient heterogeneity of non-photochemical quenching in shade-grown heterobaric leaves of avocado (Persea americana L.): responses to CO2 concentration, stomatal occlusion, dehydration and relative humidity.

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Takayama, Kotaro; King, Diana; Robinson, Sharon A; Osmond, Barry

2013-11-01

Long-lived shade leaves of avocado had extremely low rates of photosynthesis. Gas exchange measurements of photosynthesis were of limited use, so we resorted to Chl fluorescence imaging (CFI) and spot measurements to evaluate photosynthetic electron transport rates (ETRs) and non-photochemical quenching (NPQ). Imaging revealed a remarkable transient heterogeneity of NPQ during photosynthetic induction in these hypostomatous, heterobaric leaves, but was adequately integrated by spot measurements, despite long-lasting artifacts from repeated saturating flashes during assays. Major veins (mid-vein, first- and second-order veins) defined areas of more static large-scale heterogeneous NPQ, with more dynamic small-scale heterogeneity most strongly expressed in mesophyll cells between third- and fourth-order veins. Both responded to external CO2 concentration ([CO2]), occlusion of stomata with Vaseline™, leaf dehydration and relative humidity (RH). We interpreted these responses in terms of independent behavior of stomata in adjacent areoles that was largely expressed through CO2-limited photosynthesis. Heterogeneity was most pronounced and prolonged in the absence of net CO2 fixation in 100 p.p.m. [CO2] when respiratory and photorespiratory CO2 cycling constrained the inferred ETR to ~75% of values in 400 or 700 p.p.m. [CO2]. Likewise, sustained higher NPQ under Vaseline™, after dehydration or at low RH, also restricted ETR to ~75% of control values. Low NPQ in chloroplast-containing cells adjacent to major veins but remote from stomata suggested internal sources of high [CO2] in these tissues.