Sample records for amyloplasts

  1. Amyloplast Sedimentation Kinetics in Corn Roots (United States)

    Leopold, A. C.; Sack, F.


    Knowledge of the parameters of amyloplast sedimentation is crucial for an evaluation of proposed mechanisms of root graviperception. Early estimates of the rate of root amyloplast sedimentation were as low as 1.2 micron/min which may be too slow for many amyloplasts to reach the vicinity of the new lower wall within the presentation time. On this basis, Haberlandt's classical statolith hypothesis involving amyloplast stimulation of a sensitive surface near the new lower wall was questioned. The aim was to determine the kinetics of amyloplast sedimentation with reference to the presentation time in living and fixed corn rootcap cells as compared with coleoptiles of the same variety.

  2. Amyloplast Distribution Directs a Root Gravitropic Reaction (United States)

    Kordyum, Elizabeth

    Immobile higher plants are oriented in the gravitational field due to gravitropim that is a physiological growth reaction and consists of three phases: reception of a gravitational signal by statocytes, its transduction to the elongation zone, and finally the organ bending. As it is known, roots are characterized with positive gravitropism, i. e. they grow in the direction of a gravitational vector, stems - with negative gravitropism, i. e. they grow in the direction opposite to a gravitational vector. According to the Nemec’s and Haberlandt’s starch-statolith hypothesis, amyloplasts in diameter of 1.5 - 3 μ in average, which appear to act as gravity sensors and fulfill a statolythic function in the specialized graviperceptive cells - statocytes, sediment in the direction of a gravitational vector in the distal part of a cell, while a nucleus is in the proximal one. There are reasonable data that confirm the amyloplasts-statoliths participation in gravity perception: 1) correlation between the statoliths localization and the site of gravity sensing, 2) significant redistribution (sedimentation) of amyloplasts in statocytes under gravistimulation in comparison with other cell organelles, 3) root decreased ability to react on gravity under starch removal from amyloplasts, 4) starchless Arabidopsis thaliana mutants are agravitropic, 5) amyloplasts-statoliths do not sediment in the absence of the gravitational vector and are in different parts or more concentrated in the center of statocytes. Plant tropisms have been intensively studied for many decades and continue to be investigated. Nevertheless, the mechanisms by which plants do so is still not clearly explained and many questions on gravisensing and graviresponse remain unanswered. Even accepted hypotheses are now being questioned and recent data are critically evaluated. Although the available data show the Ca2+ and cytoskeleton participation in graviperception and signal transduction, the clear evidence

  3. Amyloplast movement and gravityperception in Arabidopsis endoderm (United States)

    Tasaka, M.; Saito, T.; Morita, M. T.

    Gravitropism of higher plant is a growth response regulating the orientation of organs elongation, which includes four sequential steps, the perception of gravistimulus, transduction of the physical stimulus to chemical signal, transmission of the signal, and differential cell elongation depending on the signal. To elucidate the molecular mechanism of these steps, we have isolated a number of Arabidopsis mutants with abnormal shoot gravitropic response. zig (zigzag)/sgr4(shoot gravitropism 4) shows little gravitropism in their shoots. Besides, their inflorescence stems elongate in a zigzag-fashion to bend at each node. ZIG encodes a SNARE, AtVTI11. sgr3 with reduced gravitropic response in inflorescence stems had a missense mutation in other SNARE, AtVAM3. These two SNAREs make a complex in the shoot endoderm cells that are gravity-sensing cells, suggesting that the vesicle transport from trans-Golgi network (TGN) to prevacuolar compartment (PVC) and/or vacuole is involved in gravitropism. Abnormal vesicular/vacuolar structures were observed in several tissues of both mutants. Moreover, SGR2 encodes phospholipase A1-like protein that resides in the vacuolar membrane. Endodermis-specific expression of these genes could complement gravitropism in each mutant. In addition, amyloplasts thought to be statoliths localized abnormally in their endoderm cells. These results strongly suggest that formation and function of vacuole in the endoderm cells are important for amyloplasts sedimentation, which is involved in the early process of shoot gravitropism. To reveal this, we constructed vertical stage microscope system to visualize the behavior of amyloplasts and vacuolar membrane in living endodermal cells. We hope to discuss the mechanism of gravity perception after showing their movements.

  4. Major Proteins of the Amyloplast of Agar and Soil - Grown Potato Tubers

    DEFF Research Database (Denmark)

    Hald, Simon; Blennow, Andreas; Stensballe, Allan;

    In potato (Solanum tuberosum) tuber starch is synthesized and stored in amyloplasts. Amyloplasts were prepared from in vitro or agar-grown micro tubers and from soil-grown mini tubers and subjected to proteome analysis. The quality of amyloplasts was assessed by comparing amyloplast fractions...

  5. Gravisensing: Ionic responses, cytoskeleton and amyloplast behavior (United States)

    Allen, N.; Chattaraj, P.; Collings, D.; Johannes, E.

    In Zea mays L., changes in orientation of stems are perceived by the pulvinal tissue, which responds to the stimulus by differential growth resulting in upward bending of the stem. Gravity is perceived in the bundle sheath cells, which contain amyloplasts that sediment to the new cell base when a change in the gravity vector occurs. The mechanism by which the mechanical signal is transduced into a physiological response is so far unknown for any gravity perceiving tissue. It is hypothesized that this involves interactions of amyloplasts with the plasma membrane and/or ER via cytoskeletal elements. To gain further insights into this process we monitored amyloplast movements in response to gravistimulation In a pharmacological approach we investigated how the dynamics of plastid sedimentation are affected by actin and microtubule disrupting drugs and modifiers of cytoplasmic pH, which is a key player in early gravitropic signaling. pHc was monitored in the cells composing the maize pulvinus before and after gravistimulation. pHc changes were only apparent within the bundle sheath cells, and not in the parenchyma cells. After turning, cytoplasmic acidification was observed at the sides of the cells, whereas the cytoplasm at the base of the cells, where plastids slowly accumulated became more basic. The results suggest that pHc has an important role in the early signaling pathways of maize stem gravitropism. Dark grown caulonemal filaments of the moss Physcomitrella patens respond to gravity vector changes with a reorientation of tip growth away from the gravity vector. Microtubule distributions in tip cells were monitored over time and seen to accumulate preferentially on the lower flank of the tip filaments 30 minutes after a 90 degree turn. Using a self-referencing Ca 2 + selective ion probe, we found that growing caulonemal filaments exhibit a Ca 2 + influx at the apical dome, similar to that reported previously for other tip growing cells. However, in

  6. SGR9, a RING type E3 ligase, modulates amyloplast dynamics important for gravity sensing. (United States)

    Morita, Miyo T.; Nakamura, Moritaka; Tasaka, Masao

    Gravitropism is triggered when the directional change of gravity is sensed in the specific cells, called statocytes. In higher plants, statocytes contain sinking heavier amyloplasts which are particular plastids accumulating starch granules. The displacement of amyloplasts within the statocytes is thought to be the initial event of gravity perception. We have demonstrated that endodermal cells are most likely to be the statocytes in Arabidop-sis shoots. Live cell imaging of the endodermal cell of stem has shown that most amyloplasts are sediment to the direction of gravity but they are not static. Several amyloplasts move dynamically in an actin filament (F-actin) dependent manner. In the presence of actin poly-merization inhibitor, all amyloplasts become static and sediment to the direction of gravity. In addition, stems treated with the inhibitor can exhibit gravitropism. These results suggest that F-actin-dependent dynamic movement of amyloplasts is not essential for gravity sensing. sgr (shoot gravitropism) 9 mutant exhibits greatly reduced shoot gravitropism. In endodermal cells of sgr9, dynamic amyloplast movement was predominantly observed and amyloplasts did not sediment to the direction of gravity. Interestingly, inhibition of actin polymerization re-stored both gravitropism and amyloplast sedimentation in sgr9. The SGR9 encodes a novel RING finger protein, which is localized to amyloplasts in endodermal cells. SGR9 showed ubiq-uitin E3 ligase activity in vitro. Together with live cell imaging of amyloplasts and F-actin, our data suggest that SGR9 modulate interaction between amyloplasts and F-actin on amylo-plasts. SGR9 positively act on amyloplasts sedimentation, probably by releasing amyloplasts from F-actin. SGR9 that is localized to amyloplast, possibly degrades unknown substrates by its E3 ligase activity, and this might promote release of amyloplasts from F-actin.

  7. Metabolic Pathways of the Wheat (Triticum aestivum)Endosperm Amyloplast Revealed by Proteomics (United States)

    By definition, amyloplasts are plastids specialized for starch production. However, amyloplasts have a broader range of functions that are not widely recognized. A recent proteomic study of amyloplasts isolated from wheat (Triticum aestivum Butte 86) endosperm suggested that they share many of the ...

  8. Pea amyloplast DNA is qualitatively similar to pea chloroplast DNA (United States)

    Gaynor, J. J.


    Amyloplast DNA (apDNA), when subjected to digestion with restriction endonucleases, yields patterns nearly identical to that of DNA from mature pea chloroplasts (ctDNA). Southern transfers of apDNA and ctDNA, probed with the large subunit (LS) gene of ribulose-1,5-bisphosphate carboxylase (Rubisco), shows hybridization to the expected restriction fragments for both apDNA and ctDNA. However, Northern transfers of total RNA from chloroplasts and amyloplasts, probed again with the LS gene of Rubisco, shows that no detectable LS meggage is found in amyloplasts although LS expression in mature chloroplasts is high. Likewise, two dimensional polyacrylamide gel electrophoresis of etiolated gravisensitive pea tissue shows that both large and small subunits of Rubisco are conspicuously absent; however, in greening tissue these two constitute the major soluble proteins. These findings suggest that although the informational content of these two organelle types is equivalent, gene expression is quite different and is presumably under nuclear control.

  9. The influence of gravity on the formation of amyloplasts in columella cells of Zea mays L (United States)

    Moore, R.; Fondren, W. M.; Koon, E. C.; Wang, C. L.


    Columella (i.e., putative graviperceptive) cells of Zea mays seedlings grown in the microgravity of outer space allocate significantly less volume to putative statoliths (amyloplasts) than do columella cells of Earth-grown seedlings. Amyloplasts of flight-grown seedlings are significantly smaller than those of ground controls, as is the average volume of individual starch grains. Similarly, the relative volume of starch in amyloplasts in columella cells of flight-grown seedlings is significantly less than that of Earth-grown seedlings. Microgravity does not significantly alter the volume of columella cells, the average number of amyloplasts per columella cell, or the number of starch grains per amyloplast. These results are discussed relative to the influence of gravity on cellular and organellar structure.

  10. Analysis of amyloplast dynamics involved in gravity sensing using a novel centrifuge microscope (United States)

    Toyota, Masatsugu; Tasaka, Masao; Morita, Miyo T.

    Plants sense gravity and change their growth orientation, a phenomenon known as gravitropism. According to the starch-statolith hypothesis, sedimentation of high-density starch-filled plastids (amyloplasts) within endodermal cells appears to be involved in gravity sensing of Arabidop-sis shoots. Recent studies suggest, however, that amyloplasts are never static but continu-ously show dynamic and complicated movements due to interaction with vacuole/cytoskeleton. Therefore, it remains unclear what movement/state of amyloplasts is required for gravity sens-ing. To address this critical issue, we analyzed gravitropism and amyloplast dynamics under hypergravity condition where sedimentation by gravity is more dominant than other movements. Segments of Arabidopsis inflorescence stem showed a gravitropism in response to hypergrav-ity (10g) that had been applied perpendicularly to the growth axis for 30 s in a conventional centrifuge, suggesting that amyloplast dynamics during this short time period is involved in gravity sensing. Real-time imaging of amyloplasts during the 10g stimulation was performed using a novel centrifuge microscope (NSK Ltd, Japan): all optical devices including objective lens, light source (LED) and CCD camera are mounted on an AC motor, enabling bright-field imaging with a temporal resolution of 30 frames/sec during rotation. Almost all amyloplasts started to move toward 10g and some reached the one side of endodermal cell within 30 s. These results clearly support the starch-statolith hypothesis that redistribution of amyloplasts to gravity is important for gravity sensing. Furthermore, we analyzed the shoot gravitropic mutant, sgr2, that has non-sedimentable amyloplasts and shows little gravitropism at 1g. An obvious gravitropism was induced by 30g for 5 min where amyloplasts were moved to the hyper-gravity but not by 10g where amyloplasts were not moved. These results not only suggest that gravity sensing of Arabidopsis inflorescence stems is

  11. Metabolic pathways of the wheat (Triticum aestivum endosperm amyloplast revealed by proteomics

    Directory of Open Access Journals (Sweden)

    Dupont Frances M


    Full Text Available Abstract Background By definition, amyloplasts are plastids specialized for starch production. However, a proteomic study of amyloplasts isolated from wheat (Triticum aestivum Butte 86 endosperm at 10 days after anthesis (DPA detected enzymes from many other metabolic and biosynthetic pathways. To better understand the role of amyloplasts in food production, the data from that study were evaluated in detail and an amyloplast metabolic map was outlined. Results Analysis of 288 proteins detected in an amyloplast preparation predicted that 178 were amyloplast proteins. Criteria included homology with known plastid proteins, prediction of a plastid transit peptide for the wheat gene product or a close homolog, known plastid location of the pathway, and predicted plastid location for other members of the same pathway. Of these, 135 enzymes were arranged into 18 pathways for carbohydrate, lipid, amino acid, nucleic acid and other biosynthetic processes that are critical for grain-fill. Functions of the other proteins are also discussed. Conclusion The pathways outlined in this paper suggest that amyloplasts play a central role in endosperm metabolism. The interacting effects of genetics and environment on starch and protein production may be mediated in part by regulatory mechanisms within this organelle.

  12. An active role of the amyloplasts and nuclei of root statocytes in graviperception (United States)

    Kordyum, E.; Guikema, J.


    Three main phases are discerned in the gravitropic reaction: perception of a gravitational stimulus, its transduction, and fixation of the reaction resulting in bending of an organ. According to the starch-statolith hypothesis of Nemec and Haberlandt, amyloplasts in the structurally and functionally specialized graviperceptive cells (statocytes) sediment in the direction of a gravitational vector in the distal part of a cell while a nucleus is in the proximal one. If amyloplasts appear to act as gravity sensors, the receptors, which interact with sedimented amyloplasts, and next signaling are still unclear. An analysis of the structural-functional organization of cells in different root cap layers of such higher plants as pea, Arabidopsis thaliana, and Brassica rapa grown under 1 g, on the clinostats, and in microgravity, allows us to support the hypothesis that amyloplasts function as statoliths in statocytes, but they may not be only the passive statolithic mass. We propose that amyloplasts fulfill a more complex function by interacting with a receptor, which is a nucleus, in transduction of some signal to it. Gravity-induced statolith movement in certain order leads to a new functional connection between gravity susceptors--amyloplasts and a receptor--a nucleus receiving some signal presumedly of a mechanical or biochemical nature from the amyloplasts. During gravitropism, sugar signaling could induce expression of genes encoding auxin transport proteins in a nucleus giving the nucleus an intermediate role in signal trunsduction following perception. c 2001 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

  13. Frequency analysis of amyloplast movement in Arabidopsis suggests dynamic gravity sensing (United States)

    Hasenstein, K.; Gilroy, S.

    We investigated the motility of amyloplasts in root caps of Arabidopsis thaliana to analyze the interaction between the acto-myosin system and plastids In addition to sedimentation cytoskeletal activity appears to lift amyloplasts and may cause impinging of sedimenting amyloplasts onto the membrane system This activity may represent graviperception We analyzed the frequency of amyloplast motion by Fourier analysis based upon the position of amyloplasts in image sequences taken at 6 second intervals The frequency analysis showed a maximum at an average cycle of about 16 seconds 0 06 Hz for vertical and lateral displacement However amyloplast velocity was frequency independent Application of the actin-depolymerizing agent Latrunculin B 0 1 mu M reduced the maximum displacement but did not change the frequency DMSO-treated roots showed a similar reduction The frequency dependency was similar for cell from different positions within the root cap The data suggest that the saltatory motion of amyloplasts is a continuous process that is mediated by cytoskeletal events and is an integral part of gravisensing Therefore gravisensing may depend on the dynamic motion of statoliths rather than simple aggregation onto the lower cell membrane Supported by NASA grants NAG2-1423 NAG10-0190 KHH and NAG2-1594 NSF MCB 02-12099 SG

  14. Gravity-Induced Amyloplast Motion in Hypocotyl and Root Statocytes of Cress Seedlings (United States)

    Svegzdiene, D.; Rakleviciene, D.; Koryzeiene, D.


    Amyloplast motion was studied by the analysis of plastid positioning in hypocotyl endodermal and root columella cells of cress seedlings during a subsequent 6-min period of gravitropic stimulation at 90°. As compared with root statocytes, a more rapid sedimentation of amyloplasts was determined in endodermal cells with simultaneous sliding towards cell centre by comparable rate within the first minute. During the second minute, the plastids remained almost in the same position in hypocotyl statocytes while they shifted slightly downwards continuing intensively to slide along root ones. After the 6 min, their shift in the both directions was more pronounced in roots than in hypocotyls. The data allow that amyloplast motion is affected by the gravity in common with the elastic cytoskeleton forces, which pull actively the plastids along the cells of the both organs. However, a quantitative relation of these forces in hypocotyl and root statocytes differs significantly during the two first minutes of gravitropic stimulation.

  15. Phosphorylation of an envelope-associated Hsp70 homolog in amyloplasts isolated from cultured cells of sycamore (Acer pseudoplatanus L.). (United States)

    Checa, S K; Viale, A M


    The presence of Hsp70 and Hsp60 molecular chaperones in amyloplasts isolated from cultured sycamore cells was analyzed by immunoblotting. Hsp70 homologs were located in both amyloplast envelope and stromal fractions, but no Hsp60 homologs were detected in any of the different suborganellar fractions. Incubation of whole amyloplasts or their envelope fraction with Mg2+ gamma-32P-ATP resulted in a rapid phosphorylation of the envelope-associated Hsp70 homolog, which constitutes a major target of phosphorylation in these plastids.

  16. Sycamore amyloplasts can import and process precursors of nuclear encoded chloroplast proteins. (United States)

    Strzalka, K; Ngernprasirtsiri, J; Watanabe, A; Akazawa, T


    Amyloplasts isolated from white-wild suspension-cultured cells of sycamore (Acer pseudoplatanus L.) are found to import and process the precursor of the small subunit (pS) of ribulose-1,5-bisphosphate carboxylase/oxygenase of spinach, but they lack the ability to form its holoenzyme due to the absence of both the large subunit and its binding-protein. They also import the precursor of the 33-kDa extrinsic protein (p33-kDa) of the O2-evolving complex of Photosystem II from spinach, but process is only to an intermediate form (i33-kDa). Chloroplasts from green-mutant cells of sycamore process p33-kDa to its mature form in this heterologous system. These results suggest that the thylakoid-associated protease responsible for the second processing step of p33-kDa is missing in amyloplasts, possibly due to the absence of thylakoid-membranes. In contrast, the apparent import of the precursor of the light-harvesting chlorophyll a/b-binding apoprotein (pLHCP) from spinach was not detected. Sycamore amyloplasts may lack the ability to import this particular thylakoid-protein, or rapidly degrade the imported molecules in the absence of thylakoid-membranes for their proper insertion.

  17. Expression of Amyloplast and Chloroplast DNA in Suspension-Cultured Cells of Sycamore (Acer pseudoplatanus L.). (United States)

    Ngernprasirtsiri, J; Macherel, D; Kobayashi, H; Akazawa, T


    Green mutant cells of sycamore (Acer pseudoplatanus L.), which had been selected by mutagenic treatment of the white wild type, grow photoheterotrophically in auxin-depleted culture medium. In contrast to the wild-type cells, mutant cells exhibit photosynthetic O(2)-evolution activity during their growth coincident with increases of (a) chlorophyll, (b) protein, and (c) ribulose-1,5-bisphosphate (RuBP) carboxylase activity. Functionally competent chloroplasts were isolated from the green cells. Mechanism(s) governing gene expression of amyloplast DNA in the heterotrophically grown white cells were compared with those of the chloroplast DNA isolated from the mutant cells. We have demonstrated in both amyloplast and chloroplast DNAs the presence of sequences homologous to the maize chloroplast genes for photosynthesis, including the large subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO)(rbcL), the 32 kDa Q(B) protein (PG32) (psbA), the apoprotein of P700 (psaA) and subunits of CF(1) (atpA, atpB, and atpE). However, employing either enzyme assays or immunological techniques, RuBisCO and CF(1) cannot be detected in the white wild type cells. Northern blot hybridization of the RNA from the white cells showed high levels of transcripts for the 16S rRNA gene and low level of transcripts for psbA; based on comparison with results obtained using the green mutant cells, we propose that the amyloplast genome is mostly inactive except for the 16S rRNA gene and psbA which is presumably regulated at the transcriptional level.

  18. Characterization and Intraorganellar Distribution of Protein Kinases in Amyloplasts Isolated from Cultured Cells of Sycamore (Acer pseudoplatanus L.). (United States)

    Viale, A M; Ngernprasirtsiri, J; Akazawa, T


    Incubation of amyloplasts isolated from cultured cells of sycamore (Acer pseudoplatanus L.) with [gamma-(32)P]ATP resulted in the rapid phosphorylation (half-time of 40 seconds at 25 degrees Celcius) of organellar polypeptides. The preferred substrate for amyloplast protein kinases was Mg(2+). ATP, and recovery of only [(32)P]serine after partial acid hydrolysis indicated the predominance of protein serine kinases in the organelle. These activities were located in the envelope and stromal fractions of the plastid, which showed different specificities toward exogenous protein substrates and distinct patterns of phosphorylation of endogenous polypeptides. A 66-kilodalton polypeptide, inaccessible to an exogenously added protease, was one of the major phosphorylated products found in intact amyloplasts at low [gamma-(32)P] adenosine triphosphate concentrations. This polypeptide represented the major phosphoprotein observed with the isolated envelope fraction. The patterns of polypeptide phosphorylation found in intact amyloplasts and chloroplasts from cultured cell lines of sycamore were clearly distinguishable. The overall results indicate the presence of protein phosphorylation systems unique to this reserve plastid present in nonphotosynthetic tissues.

  19. Amyloplasts are necessary for full gravitropic sensitivity in roots of Arabidopsis thaliana (United States)

    Kiss, J. Z.; Hertel, R.; Sack, F. D.


    The observation that a starchless mutant (TC7) of Arabidopsis thaliana (L.) Heynh. is gravitropic (T. Caspar and B.G. Pickard, 1989, Planta 177, 185-197) raises questions about the hypothesis that starch and amyloplasts play a role in gravity perception. We compared the kinetics of gravitropism in this starchless mutant and the wild-type (WT). Wild-type roots are more responsive to gravity than TC7 roots as judged by several parameters: (1) Vertically grown TC7 roots were not as oriented with respect to the gravity vector as WT roots. (2) In the time course of curvature after gravistimulation, curvature in TC7 roots was delayed and reduced compared to WT roots. (3) TC7 roots curved less than WT roots following a single, short (induction) period of gravistimulation, and WT, but not TC7, roots curved in response to a 1-min period of horizontal exposure. (4) Wild-type roots curved much more than TC7 roots in response to intermittent stimulation (repeated short periods of horizontal exposure); WT roots curved in response to 10 s of stimulation or less, but TC7 roots required 2 min of stimulation to produce a curvature. The growth rates were equal for both genotypes. We conclude that WT roots are more sensitive to gravity than TC7 roots. Starch is not required for gravity perception in TC7 roots, but is necessary for full sensitivity; thus it is likely that amyloplasts function as statoliths in WT Arabidopsis roots. Furthermore, since centrifugation studies using low gravitational forces indicated that starchless plastids are relatively dense and are the most movable component in TC7 columella cells, the starchless plastids may also function as statoliths.

  20. DNA methylation is a determinative element of photosynthesis gene expression in amyloplasts from liquid-cultured cells of sycamore (Acer pseudoplatanus L.). (United States)

    Ngernprasirtsiri, J; Kobayashi, H; Akazawa, T


    Transcriptional regulation has been shown to operate as a selective control mechanism of expression of photosynthetic genes in the nonphotosynthetic plastids, amyloplasts, of a white-wild cell line of sycamore (Acer pseudoplatanus L.). To elaborate the mechanisms governing the transcriptional regulation at the molecular level, we have examined the template activity of the amyloplast DNA compared to the chloroplast DNA by using the in vitro run-off transcription assay system with extracts of the two plastid types. The results of these assays clearly indicate that most of the amyloplast DNA regions do not serve as a template for the in vitro transcription regardless of the plastid extracts; this is in contrast to the chloroplast DNA which serves as an active template. It is highly likely that the template activity of amyloplast DNA per se is the modulating element of transcriptional regulation. Parallel experiments determining the DNA base content by HPLC analysis have shown that a variety of methylated bases, especially 5-methylcytosine, are localized in the DNA regions containing suppressed genes of the amyloplast genome. In sharp contrast, methylated bases were undetectable in the expressed gene regions of amyloplast and whole chloroplast genomes. The overall findings strongly support the notion that DNA methylation is involved in the selective suppression of photosynthetic genes in the nonphotosynthetic plastids of cultured sycamore cells.

  1. Variation in stem morphology and movement of amyloplasts in white spruce grown in the weightless environment of the International Space Station. (United States)

    Rioux, Danny; Lagacé, Marie; Cohen, Luchino Y; Beaulieu, Jean


    One-year-old white spruce (Picea glauca) seedlings were studied in microgravity conditions in the International Space Station (ISS) and compared with seedlings grown on Earth. Leaf growth was clearly stimulated in space whereas data suggest a similar trend for the shoots. Needles on the current shoots of ground-based seedlings were more inclined towards the stem base than those of seedlings grown in the ISS. Amyloplasts sedimented in specialized cells of shoots and roots in seedlings grown on Earth while they were distributed at random in similar cells of seedlings tested in the ISS. In shoots, such amyloplasts were found in starch sheath cells located between leaf traces and cortical cells whereas in roots they were constituents of columella cells of the cap. Nuclei were regularly observed just above the sedimented amyloplasts in both organs. It was also frequent to detect vacuoles with phenolic compounds and endoplasmic reticulum (ER) close to the sedimented amyloplasts. The ER was mainly observed just under these amyloplasts. Thus, when amyloplasts sediment, the pressure exerted on the ER, the organelle that can for instance secrete proteins destined for the plasma membrane, might influence their functioning and play a role in signaling pathways involved in gravity-sensing white spruce cells.

  2. High-capacity calcium-binding chitinase III from pomegranate seeds (Punica granatum Linn.) is located in amyloplasts


    Lv, Chenyan; Masuda, Taro; Yang, Haixia; Sun, Lei; Zhao, Guanghua


    We have recently identified a new class III chitinase from pomegranate seeds (PSC). Interestingly, this new chitinase naturally binds calcium ions with high capacity and low affinity, suggesting that PSC is a Ca-storage protein. Analysis of the amino acid sequence showed that this enzyme is rich in acidic amino acid residues, especially Asp, which are responsible for calcium binding. Different from other known chitinases, PSC is located in the stroma of amyloplasts in pomegranate seeds. Trans...

  3. Application of an efficient strategy with a phage lambda vector for constructing a physical map of the amyloplast genome of sycamore (Acer pseudoplatanus). (United States)

    Ngernprasirtsiri, J; Kobayashi, H


    Amyloplasts were isolated from a heterotrophic culture cell line of a woody plant, sycamore (Acer pseudoplatanus), and their DNA was purified. Conventional procedures for making a physical map were not easily applicable to the amyloplast DNA, since the yield of DNA was too low and the presence of repeated sequences interfered with the analysis. Therefore, the pieces of amyloplast DNA starting with a few micrograms of DNA were cloned in the lambda Fix vector, which is a derivative of lambda EMBL vectors improved for efficient cloning and gene walking. Cloned DNA fragments were randomly picked, mapped for restriction endonuclease sites by a refined procedure, and combined by overlapping their physical maps. The DNA library was also subjected to screening by gene walking using promoters recognized by T3 and T7 RNA polymerases in the vector to fill the gaps between sequences determined by overlapping the physical maps. In this way, we constructed the entire DNA library and the complete physical map of the amyloplast DNA. The sycamore amyloplast genome was composed of 141.7-kbp nucleotides with the same gene arrangement as that of tobacco chloroplasts.

  4. Amyloplast to chromoplast conversion in developing ornamental tobacco floral nectaries provides sugar for nectar and antioxidants for protection. (United States)

    Horner, H T; Healy, R A; Ren, G; Fritz, D; Klyne, A; Seames, C; Thornburg, R W


    Tobacco floral nectaries undergo changes in form and function. As nectaries change from green to orange, a new pigment is expressed. Analysis demonstrated that it is β-carotene. Plastids undergo dramatic changes. Early in nectary development, they divide and by stage 9 (S9) they are engorged with starch. About S9, nectaries shift from quiescent anabolism to active catabolism resulting in starch breakdown and production of nectar sugars. Starch is replaced by osmiophilic bodies, which contain needle-like carotenoid crystals. Between S9 and S12, amyloplasts are converted to chromoplasts. Changes in carotenoids and ascorbate were assayed and are expressed at low levels early in development; however, following S9 metabolic shift, syntheses of β-carotene and ascorbate greatly increase in advance of expression of nectar redox cycle. Transcript analysis for carotenoid and ascorbate biosynthetic pathways showed that these genes are significantly expressed at S6, prior to the S9 metabolic shift. Thus, formation of antioxidants β-carotene and ascorbate after the metabolic shift is independent of transcriptional regulation. We propose that biosynthesis of these antioxidants is governed by availability of substrate molecules that arise from starch breakdown. These processes and events may be amenable to molecular manipulation to provide a better system for insect attraction, cross pollination, and hybridization.

  5. Isolation of cDNA encoding a newly identified major allergenic protein of rye-grass pollen: intracellular targeting to the amyloplast.


    Singh, M. B.; Hough, T.; Theerakulpisut, P; Avjioglu, A; S. Davies; Smith, P. M.; Taylor, P.; Simpson, R. J.; Ward, L. D.; McCluskey, J


    We have identified a major allergenic protein from rye-grass pollen, tentatively designated Lol pIb of 31kDa and with pI 9.0. A cDNA clone encoding Lol pIb has been isolated, sequenced, and characterized. Lol pIb is located mainly in the starch granules. This is a distinct allergen from Lol pI, which is located in the cytosol. Lol pIb is synthesized in pollen as a pre-allergen with a transit peptide targeting the allergen to amyloplasts. Epitope mapping of the fusion protein localized the IgE...

  6. Kinetics of gravity-induced amyloplast sedimentation in statocytes of cress roots grown under fast clino-rotation, 1 g and after 180° inversion (United States)

    Svegzdiene, D.; Rakleviciene, D.; Gaina, V.

    In order to detail the relation between the initial positioning of statoliths and root graviresponsiveness, the movement of amyloplasts within root statocytes of cress ( Lepidium sativum L.) seedlings grown under fast clino-rotation (50 rpm) and vertically at 1 g or 180° inverted after growth at 1 g has been analyzed during a subsequent 6-min period of lateral (reoriented 90°) gravistimulation. The main parameters of statolith motion kinetics such as relative positions and motion velocities in transverse and longitudinal directions have been evaluated taking linear measurements of organelle position with respect to the lower longitudinal and distal wall in the cells of the 2nd-5th columella storeys. A significant discrepancy in the direction and velocity of statolith movement has been determined within the first and second minutes of root horizontal placement. Centrally or proximally located statoliths in clino-rotated or inverted root statocytes moved transversely to the lower longitudinal cell wall more quickly as compared with distally located ones in 1- g root cells. At the same time, the longitudinal displacement of proximally located statoliths towards the cell center has been determined, while this displacement of distally located plastids was significantly more pronounced. The longitudinal motion of amyloplasts grouped in the central cell part was negligible within this 2-min period. After the 6th minute of stimulation, the total downward shift of the mean relative position of statoliths amounted to 16.3% from the total cell width in clino-rotated root statocytes and 17.1% in the cells of inverted roots, while it equalled 13.3% in 1- g root statocytes. The total change of the relative longitudinal position of amyloplasts with respect to the distal cell wall remained almost unchanged in roots grown on the clinostat. In inverted roots, the initial longitudinal position of 56.2% from the total cell length has been shifted significantly to 47.8% and from 27

  7. Expression of photosynthetic genes is distinctly different between chloroplasts and amyloplasts in the liquid-cultured cells of sycamore (Acer pseudoplatanus L.). (United States)

    Ngernprasirtsiri, J; Kobayashi, H; Akazawa, T


    A nonphotosynthetic, white-wild cell line of sycamore (Acer pseudoplatanus L.) contains amyloplasts as the only kind of plastid, whereas a photosynthetically competent green variant cell line contains only chloroplasts. Transcripts of both nuclear and plastid genes for photosynthetic components in the white cells were not detectable in contrast to those in the green cells. To investigate the limiting step (s) behind these diminished levels of transcripts, we have performed in vivo pulse-chase labeling of RNA in both cell types. These studies indicated that the rates of incorporation of [3H]uridine and nucleotide pool sizes were indistinguishable between the two cell lines. Transcripts of certain nuclear (rbcS, cab, psbO) and plastid (rbcL) genes in the white cell were not detectable. We infer from these data that transcriptional regulation entails an important role in controlling photosynthetic RNA levels. Related analyses exploiting plastid run-on transcription have provided supporting evidence that the transcription of the amyloplast genome in the white cell is greatly suppressed in contrast to that of the chloroplast genome in the green cell. The results support a model of selective suppression of photosynthesis genes in nonphotosynthetic higher plant cells, and indicate that gene expression in such a system is primarily controlled at the transcriptional level.

  8. Identification of a homolog of Arabidopsis DSP4 (SEX4) in chestnut: its induction and accumulation in stem amyloplasts during winter or in response to the cold. (United States)

    Berrocal-Lobo, Marta; Ibañez, Cristian; Acebo, Paloma; Ramos, Alberto; Perez-Solis, Estefania; Collada, Carmen; Casado, Rosa; Aragoncillo, Cipriano; Allona, Isabel


    Oligosaccharide synthesis is an important cryoprotection strategy used by woody plants during winter dormancy. At the onset of autumn, starch stored in the stem and buds is broken down in response to the shorter days and lower temperatures resulting in the buildup of oligosaccharides. Given that the enzyme DSP4 is necessary for diurnal starch degradation in Arabidopsis leaves, this study was designed to address the role of DSP4 in this seasonal process in Castanea sativa Mill. The expression pattern of the CsDSP4 gene in cells of the chestnut stem was found to parallel starch catabolism. In this organ, DSP4 protein levels started to rise at the start of autumn and elevated levels persisted until the onset of spring. In addition, exposure of chestnut plantlets to 4 °C induced the expression of the CsDSP4 gene. In dormant trees or cold-stressed plantlets, the CsDSP4 protein was immunolocalized both in the amyloplast stroma and nucleus of stem cells, whereas in the conditions of vegetative growth, immunofluorescence was only detected in the nucleus. The studies indicate a potential role for DSP4 in starch degradation and cold acclimation following low temperature exposure during activity-dormancy transition.

  9. Kinetics of Gravity-induced Amyloplast Sedimentation in Cress Root Statocytes Formed under 1 g and on Fast-rotating Clinostat (United States)

    Svegzdiene, D.; Rakleviciene, D.; Gaina, V.

    The aim of the sudy was to compare the motion of a statolith complex in statocytes of roots grown vertically under 1 g and simulated weightlessness by fast-clinorotation (50 rpm) during lateral stimulation by the gravity vector; the experiments with cress (Lepidium sativum L.) on a centrifuge-clinostat device have been performed. Before gravistimulation, the statoliths are grouped in the distal region of 1g-root statocytes and at the center of statocytes in clino-rotated roots. Then roots were placed horizontally for increasing periods of time (1, 2, 4 or 6 min) and chemically fixed. Quantitative analysis of amyloplast movement in side-to-side and distal-to-proximal directions within statocytes of the 2nd to 5th statenchyma storeys was accomplished by light and electron microscopy. After the first minute of gravistimulation, the position of statolith complex in the 1g-statocytes has been changed by about 12.2% in side-to-side and 18.3% in distal-to-proximal direction versus its initial position. In roots grown on the clinostat, the plastids changed their initial position by 22.5% toward the lower longitudinal wall of the statocyte and negligibly (2.4%) -- toward the proximal cell wall during this stimulation period. Later, up to 2 min, the statoliths continued to displace following the alike trajectory in side-to-side by 36.6% as well as in distal-to-proximal direction by 34.4% in 1g-root statocytes versus their position occupied at 1 min, while they remained approximately at the same position in statocytes of clinostat-grown roots. During the period from 2 to 6 min, in the former roots a significant further shift (15.1%) parallel to the gravity vector and otherwise a reversible movement (- 13.3%) toward the distal wall of statolith complex were observed. However, within the same period of gravistimulation the statolith positioning in clinorotated-roots changed only slightly in side-to-side (4.8%) as well as in distal-to-proximal (2.1%) directions. The obtained

  10. 板栗种子淀粉体发育的扫描电镜观察%Observation of amyloplast development in chestnut seed by scanning electron microscope

    Institute of Scientific and Technical Information of China (English)

    刘帅; 陈良珂; 房克凤; 杨瑞; 邢宇; 曹庆芹; 秦岭


    In order to get the growth law of the Chinese chestnut seed amyloid, the chestnut seeds at different development stages were used as the experiment materials. The size, shape and development condition of amyloid features were observed by scanning electron microscopy. Results show that the aleurone layer cells have no amyloid and accumulation of starch. The amyloid mainly concentrates in the cotyledons. The chestnut seeds contain two kinds of amyliod:the large are pebble and the small are round. The large amyloid is the main kind, and its spatial arrangement is close. In addition, the amyliod has other forms in chestnut seeds, such as polyhedral, spherical, kidney, and complex form. The volume of large amyloid changes obviously in the prophase and metaphase stages in chestnut seeds. The small amyloplast proliferation mainly occurres in middle and late stages of seed growth, and distributes in clearance of large amyloid. As the chestnut seed develops, the volume of amyloid gradually increases, and the growth change of long axis and short axis shows “S” type in the growing curves.%为了探明板栗种子淀粉体生长的规律,以不同发育时期的板栗种子为试验材料,对淀粉体的大小、形态和发育情况等特性进行扫描电镜观察。结果表明:板栗糊粉层细胞中无淀粉体,不积累淀粉,淀粉体主要集中在子叶中;板栗种子有大、小两种淀粉体,大淀粉体呈鹅卵石形,小淀粉体呈圆球形;板栗种子以大淀粉体为主,空间排布较为紧密,此外,板栗淀粉体具有多面体形、球形、肾形、复合形等形态;板栗大淀粉体在种子发育前期和中期体积变化比较明显,而小淀粉体分布于大淀粉体的间隙中,增殖主要发生在种子发育中期和后期;淀粉体体积随发育天数的增加逐渐增大,长轴和短轴增长的变化呈“S”型生长曲线。

  11. Mutan produced in potato amyloplasts adheres to starch granules. (United States)

    Kok-Jacon, Géraldine A; Vincken, Jean-Paul; Suurs, Luc C J M; Visser, Richard G F


    Production of water-insoluble mutan polymers in Kardal potato tubers was investigated after expression of a full-length (GtfI) and a truncated mutansucrase gene referred to as GtfICAT (GtfI without glucan-binding domain) from Streptococcus downei. Subsequent effects on starch biosynthesis at the molecular and biochemical levels were studied. Expression of the GtfICAT gene resulted in the adhesion of mutan material on starch granules, which stained red with erythrosine, and which was hydrolysed by exo-mutanase. In addition, GtfICAT-expressing plants exhibited a severely altered tuber phenotype and starch granule morphology in comparison to those expressing the full-length GtfI gene. In spite of that, no structural changes at the starch level were observed. Expression levels of the sucrose-regulated, AGPase and GBSSI genes were down-regulated in only the GTFICAT transformants, showing that GtfICAT expression interfered with the starch biosynthetic pathway. In accordance with the down-regulated AGPase gene, a lower starch content was observed in the GTFICAT transformants. Finally, the rheological properties of the GTFICAT starches were modified; they showed a higher retrogradation during cooling of the starch paste. PMID:17129316

  12. Mutan produced in potato amyloplasts adheres to starch granules

    NARCIS (Netherlands)

    Kok-Jacon, G.A.; Vincken, J.P.; Suurs, L.C.J.M.; Visser, R.G.F.


    Production of water-insoluble mutan polymers in Kardal potato tubers was investigated after expression of a full-length (GtfI) and a truncated mutansucrase gene referred to as GtfICAT (GtfI without glucan-binding domain) from Streptococcus downei. Subsequent effects on starch biosynthesis at the mol

  13. Effect of the change in statocyte polarity on calcium distribution: results from PolCa space experiment (United States)

    Legue, Valerie; Pereda, Veronica; Gerard, Joelle; Eche, Brigitte; Gasset, Gilbert; Chaput, Didier

    Plants are the ability to sense and to re-orient their growth in response to gravity. In roots, specialized sensory cells (called statocytes) perceive signal gravity and are the only cells that exhibit structural polarity with respect to gravity providing interactions with starch-containing plastids (amyloplasts) and the cortical endoplasmic reticulum (ER). Upon root reorientation, a displacement of amyloplasts is observed and is accompanied with a change in direct amyloplast-ER direct interaction in root cap cells. Even if amyloplasts are widely considered as gravity sensor, there is no clear evidence that a change in amyloplasts-ER interactions could lead to a transduction gravity signal. Previous space experiments clearly showed that amyloplasts inter-action with ER are not necessary to lead to a root re-orientation, suggesting that amyloplasts displacement mediate transduction events through cytoskeleton reorganisation and calcium-dependant pathways. The objective of the space experiment called PolCa is to dissect the effect of change in amyloplasts-ER interactions on calcium dependant pathways. Space conditions provide a unique opportunity to provide a change of structural polarity in statocytes without a gravistimulation. PolCa experiment has been conducted using Brassica napus seedlings, which submitted four different conditions: continuously on 1 g centrifuge or continuously in micrograv-ity conditions. Some seedlings germinated on centrifuge have been transferred during 10 min in microgravity conditions, leading to a loss of amyloplast-ER interactions through amyloplasts displacement. Others seedlings germinated in microgravity conditions have been transferred on centrifuge during 10 min. In this situation, we observed a return of amyloplasts-ER interactions. We analysed the localisation of free calcium after chemical fixation and calcium precipitation using potassium pyroantimonate in these four situations. The observation of calcium precipita-tions with

  14. ジャガイモ塊茎におけるプラスチド-アミロプラスト系の微細構造とデンプンの合成・蓄積に関する電子顕微鏡観察


    川崎, 通夫 / 松田, 智明 / 新田, 洋司; カワサキ, ミチオ / マツダ, トシアキ / ニッタ, ヨウジ; kAWASAKI, Michio / MATSUDA, Toshiaki / NAKAMURA, Yasunori / UENO, Osamu / TANIGUCHI, Mitsutaka / NITTA, Youji / MIYAKE, Hiroshi


    The starch accumulation process and the ultrastructure of the plastid-amyloplast system in potato tuber were observed by electron and light microscopes. Amyloplasts were slightly flat ovalshaped and contained a single starch grain. Even if they were lrge, their replications were observed. In the parenchyma cells around the phloem, the plastid-amyloplast system intensively replicated and grew larger with starch accumulation. As a result, much starch was accumulated in the parenchyma cells in t...

  15. Structural and Immunocytochemical Characterization of the Synthesis and Accumulation of Starch in Sweet Potato (Ipomoea batatas Lam.) Tuberous Root


    kAWASAKI, Michio / MATSUDA, Toshiaki / NAKAMURA, Yasunori / UENO, Osamu / TANIGUCHI, Mitsutaka / NITTA, Youji / MIYAKE, Hiroshi


    The structural changes in the plastid-amyloplast system in the parenchyma cells of sweet potato tuberous roots during thickening were examined by electron microscopy. In the tuberous roots, proplastids and plastids that contain starch granules propagated in young parenchyma cells adjacent to the meristem, but amyloplasts did not in parenchyma cells. It was suggested that the number of amyloplasts in a parenchyma cell is determined by the propagation of the proplastids and plastids. The form o...


    NARCIS (Netherlands)



    Potato branching enzyme, a key enzyme in the biosynthesis of starch, was localized in amyloplasts in starch-storage cells of potato (Solanum tuberosum L) with the use of immunogold electron microscopy. Branching enzyme was found in the amyloplast stroma, concentrated at the interface of the stroma a

  17. Distinct isoforms of ADPglucose pyrophosphatase and ADPglucose pyrophosphorylase occur in the suspension-cultured cells of sycamore (Acer pseudoplatanus L.). (United States)

    Baroja-Fernández, E; Zandueta-Criado, A; Rodríguez-López, M; Akazawa, T; Pozueta-Romero, J


    The intracellular localizations of ADPglucose pyrophosphatase (AGPPase) and ADPglucose pyrophosphorylase (AGPase) have been studied using protoplasts prepared from suspension-cultured cells of sycamore (Acer pseudoplatanus L.). Subcellular fractionation studies revealed that all the AGPPase present in the protoplasts is associated with amyloplasts, whereas more than 60% of AGPase is in the extraplastidial compartment. Immunoblots of amyloplast- and extraplastid-enriched extracts further confirmed that AGPase is located mainly outside the amyloplast. Experiments carried out to identify possible different isoforms of AGPPase in the amyloplast revealed the presence of soluble and starch granule-bound isoforms. We thus propose that ADPglucose levels linked to starch biosynthesis in sycamore cells are controlled by enzymatic reactions catalyzing the synthesis and breakdown of ADPglucose, which take place both inside and outside the amyloplast.

  18. A new centrifuge microscope reveals that mobile plastids trigger gravity sensing in Arabidopsis inflorescence stems (United States)

    Toyota, Masatsugu; Tasaka, Masao; Morita, Miyo T.; Gilroy, Simon


    The starch-statolith hypothesis is the most widely accepted model for plant gravity sensing and proposes that the sedimentation of high-density starch-filled plastids (amyloplasts) in shoot endodermal cells and root columella cells is important for gravity sensing of each organ. However, starch-deficient phosphoglucomutase (pgm-1) mutants sense gravity and show gravitropism in inflorescence stems, even though most starchless amyloplasts in this mutant fail to sediment toward the gravity vector. These results raise the questions about the role of starch in gravity sensing and the features of statolith/statocyte essential for shoot gravity sensing. To address these questions, we developed a new centrifuge microscope and analyzed two gravitropic mutants, i.e., pgm-1 and endodermal-amyloplast less 1 (eal1). All optical devices (e.g., objective lens, light source and CCD camera) and specimens were rotated on a direct-drive motor, and acquired images were wirelessly transmitted during centrifugation. Live-cell imaging during centrifugation revealed that the starchless amyloplasts sedimented to the hypergravity vector (10 and 30 g) in endodermal cells of pgm-1 stems, indicating that the density of the starchless amyloplasts is higher than that of cytoplasm. Electron micrographs of shoot endodermal cells in pgm-1 mutants suggested that the starchless amyloplast contains an organized thylakoid membrane but not starch granules, which morphologically resembles chloroplasts in the adjacent cortical cells. Therefore, the shoot amyloplasts without starch are possibly as dense as chloroplasts. We examined eal1 mutants, an allele of shoot gravitropism (sgr) 7/short-root (shr), which also have starchless amyloplasts due to abnormal differentiation of amyloplasts and show no gravitropic response at 1 g. Hypergravity up to 30 g induced little gravitropism in eal1 stems and the starchless amyloplasts failed to sediment under 30 g conditions. However, the eal1 mutants treated with

  19. A morphometric analysis of the redistribution of organelles in columella cells of horizontally-oriented roots of Zea mays (United States)

    Moore, R.


    In order to determine what structural changes in graviperceptive cells are associated with onset of root gravicurvature, the redistribution of organelles in columella cells of horizontally-oriented, graviresponding roots of Zea mays has been quantified. Root gravicurvature began by 15 min after reorientation, and did not involve significant changes in the (i) volume of individual columella cells or amyloplasts, (ii) relative volume of any cellular organelle, (iii) number of amyloplasts per columella cell, or (iv) surface area of cellular location of endoplasmic reticulum. Sedimentation of amyloplasts began within 1 to 2 min after reorientation, and was characterized by an intensely staining area of cytoplasm adjacent to the sedimenting amyloplasts. By 5 min after reorientation, amyloplasts were located in the lower distal corner of columella cells, and, by 15 min after reorientation, overlaid the entire length of the lower cell wall. No consistent contact between amyloplasts and any cellular structure was detected at any stage of gravicurvature. Centrally-located nuclei initially migrated upward in columella cells of horizontally-oriented roots, after which they moved to the proximal ends of the cells by 15 min after reorientation. No significant pattern of redistribution of vacuoles, mitochondria, dictyosomes, or hyaloplasm was detected that correlated with the onset of gravicurvature. These results indicate that amyloplasts and nuclei are the only organelles whose movements correlate positively with the onset of gravicurvature by primary roots of this cultivar of Zea mays.

  20. The effect of 3-indolylacetic acid on the accumulation of starch in the root tissue of Cichorium intybus L. cultured in vitro

    Directory of Open Access Journals (Sweden)

    U. Sobczyk


    Full Text Available The relation between IAA-induced formation of amyloplasts in callus cells of chicory root and the influence of IAA on sugar uptake from the medium was investigated. Experiments with 14C-sucrose showed that IAA increased the uptake of sucrose from the medium. The amyloplast-like structures were also observed in callus grown on medium without IAA, but containing high concentration of sucrose (9%. The possibility of IAA influence on the formation of amyloplasts by increasing the permeability of cells for sugar is discussed.

  1. Effect of Temperature on the Decomposition of Reserves during Germination and Early Growth of Rice Plants : An electron microscopic study


    ZAKARIA, Sabaruddin / MATSUDA, Toshiaki / TAJIMA, Shingo / NITTA, Youji


    Morphological changes in starch granules in endosperm cells of rice, cv. Cisadane (javanica) and cv. Koshihikari (japonica), during germination and early growth of seedlings at 17℃, ambient temperature (field condition, 16-24℃) and 38℃, respectively, were examined with scanning and transmission electron microscopes. With the treatment at 17℃, a groove-like structure appeared on the surface of amyloplasts at five days after seeding (DAS). After the amyloplast envelope disappeared and starch gr...

  2. Caryopsis Development and Main Quality Characteristics in Different indica Rice Varieties

    Institute of Scientific and Technical Information of China (English)

    XIONG Fei; WANG Zhong; CHENG Gang; WANG Jue


    A comparison on the caryopsis development, rice quality characteristics and the size, shape, structure of the endosperm amyloplasts (starch granules) between two indica rice varieties was made. The main quality traits in Yangdao 6 were better than those in Xiangzaoxian 33; In the early period after fertilization the dry matter in Yangdao 6 was accumulated more slowly than that in Xiangzaoxian 33 but faster in the later period, and the starch was accumulated strongly in the later period; There were two kinds of amyloplasts: single and compound amyloplasts, being 4.4 μm and 9.5 μm in diameter on average with the range of 2.4-8.0 μm and 5.7-19.5 μm, respectively. In the case of Xiangzaoxian 33, most of the single amyloplasts were elliptic or round with loose arrangement and great difference in size, and the coefficient of variation was high. While in the Yangdao 6, most of the amyloplasts were single, well developed, polyhedral, crystalline and compactly arranged, and the coefficient of variation was low. The amyloplasts in the dorsal region of endosperm were developed better than those in the ventral and central regions. The chalkiness in the endosperm resulted from badly-developed and loose-arranged starch granules, which was closely relevant to the transport of filling materials.

  3. Role of endodermal cell vacuoles in shoot gravitropism. (United States)

    Kato, Takehide; Morita, Miyo Terao; Tasaka, Masao


    In higher plants, shoots and roots show negative and positive gravitropism, respectively. Data from surgical ablation experiments and analysis of starch deficient mutants have led to the suggestion that columella cells in the root cap function as gravity perception cells. On the other hand, endodermal cells are believed to be the statocytes (that is, gravity perceiving cells) of shoots. Statocytes in shoots and roots commonly contain amyloplasts which sediment under gravity. Through genetic research with Arabidopsis shoot gravitropism mutants, sgr1/scr and sgr7/shr, it was determined that endodermal cells are essential for shoot gravitropism. Moreover, some starch biosynthesis genes and EAL1 are important for the formation and maturation of amyloplasts in shoot endodermis. Thus, amyloplasts in the shoot endodermis would function as statoliths, just as in roots. The study of the sgr2 and zig/sgr4 mutants provides new insights into the early steps of shoot gravitropism, which still remains unclear. SGR2 and ZIG/SGR4 genes encode a phospholipase-like and a v-SNARE protein, respectively. Moreover, these genes are involved in vacuolar formation or function. Thus, the vacuole must play an important role in amyloplast sedimentation because the sgr2 and zig/sgr4 mutants display abnormal amyloplast sedimentation.

  4. Deficiency of Starch Synthase IIIa and IVb Alters Starch Granule Morphology from Polyhedral to Spherical in Rice Endosperm. (United States)

    Toyosawa, Yoshiko; Kawagoe, Yasushi; Matsushima, Ryo; Crofts, Naoko; Ogawa, Masahiro; Fukuda, Masako; Kumamaru, Toshihiro; Okazaki, Yozo; Kusano, Miyako; Saito, Kazuki; Toyooka, Kiminori; Sato, Mayuko; Ai, Yongfeng; Jane, Jay-Lin; Nakamura, Yasunori; Fujita, Naoko


    Starch granule morphology differs markedly among plant species. However, the mechanisms controlling starch granule morphology have not been elucidated. Rice (Oryza sativa) endosperm produces characteristic compound-type granules containing dozens of polyhedral starch granules within an amyloplast. Some other cereal species produce simple-type granules, in which only one starch granule is present per amyloplast. A double mutant rice deficient in the starch synthase (SS) genes SSIIIa and SSIVb (ss3a ss4b) produced spherical starch granules, whereas the parental single mutants produced polyhedral starch granules similar to the wild type. The ss3a ss4b amyloplasts contained compound-type starch granules during early developmental stages, and spherical granules were separated from each other during subsequent amyloplast development and seed dehydration. Analysis of glucan chain length distribution identified overlapping roles for SSIIIa and SSIVb in amylopectin chain synthesis, with a degree of polymerization of 42 or greater. Confocal fluorescence microscopy and immunoelectron microscopy of wild-type developing rice seeds revealed that the majority of SSIVb was localized between starch granules. Therefore, we propose that SSIIIa and SSIVb have crucial roles in determining starch granule morphology and in maintaining the amyloplast envelope structure. We present a model of spherical starch granule production. PMID:26747287

  5. Action of the mechanical disruption of the actin network on the gravisensitivity of the root statocyte (United States)

    Lefranc, A.; Jeune, B.; Driss-Ecole, D.; Perbal, G.

    The effects of the mechanical disruption of the thin actin network of statocytes on gravisensitivity have been studied on lentil roots. Seedling roots were first inverted for 7 min (root tip upward) and then placed in the downward (normal) position for 7 min before gravitropic stimulation in the horizontal position. The period of inversion allowed the amyloplasts to move from the distal part to the proximal part of the statocyte, but did not fully sediment. When the roots were returned to the tip down position, the amyloplasts moved toward the distal part, but also did not completely sediment by the time the roots were placed horizontally. Thus, in these roots the amyloplasts could be still moving toward the distal wall after they had been replaced in the normal position and the actin network should not be fully restored. Gravisensitivity was estimated by the analysis of the dose-response curves of vertical and treated (inverted and returned to downward position) roots. The only effect, which has been observed on treated roots, was a delay of graviresponse for about 1 min. Our interpretation of this result is that in vertical roots the amyloplasts can exert tensions in the actin network that are directly transmitted to mechanoreceptors located in the plasma membrane. In roots with a partially disrupted actin network, a delay of 1 min is necessary for the amyloplasts to activate mechanoreceptors.

  6. The Effect of Weak Combined Magnetic Field on Root Gravitropism and a Role of Ca2+ Ions Therein (United States)

    Kordyum, Elizabeth; Bogatina, Nina; Kondrachuk, A.

    At present, magnetic fields of different types are widely used to study gravity sensing in plants. For instance, magnetic levitation of amyloplasts caused by high gradient magnetic field enables us to alter the effective gravity sensed by plant cells. For the first time we showed that a weak combined magnetic field (CMF), that is the sum of collinear permanent and alternating magnetic fields ( 0.5 gauss, 0-100 Hz), changes a cress and pea root positive gravitropic reaction on a negative one. This effect has the form of resonance and occurs at the frequency of cyclotron resonance of calcium ions. What is especially interesting is that under gravistimulation in the CMF, the displacement of amylopasts in the root cap statocytes is directed to the upper wall of a cell, i.e. in the direction opposite to the gravitational vector. The displacement of amyloplasts, which contain the abundance of free Ca2+ ions in the stroma, is accompanied by Ca2+ redistribution in the same direction, and increasing in the cytosol around amyloplasts near ten times in the CMF in comparison with the state magnetic field. Earlier, we also observed the Ca2+ accumulation in the upper site of a root curvature in the elongation zone in the CMF unlike a positive gravitropic reaction. Thus, it should be stressed that a root is bending in the same direction in which amyloplasts are displacing: downwards when gravitropism is positive and upwards when gravitropism is negative. The obtained data confirm the amyloplast statolithic function and give another striking demonstration of a leading role of Ca2+ ions in root gravitropism. But these data bring the question: what forces can promote amyloplast displacement against gravity? The possible explanation of the effect found is discussed. It is based on the ion cyclotron resonance in biosystems proposed by Liboff.. The original approach based on the use of a weak CMF may be helpful for understanding the mechanisms of plant gravisensing

  7. Kinetics of starch digestion and performance of broiler chickens

    NARCIS (Netherlands)

    Weurding, R.E.


    Keywords: starch, digestion rate, broiler chickens, peas, tapiocaStarch is stored in amyloplasts of various plants like cereals and legumes and seeds of these plants are used as feedstuffs for farm animals. Starch is the major energy source in broiler feeds. The properties of star

  8. A unique HEAT repeat-containing protein SHOOT GRAVITROPISM6 is involved in vacuolar membrane dynamics in gravity-sensing cells of Arabidopsis inflorescence stem. (United States)

    Hashiguchi, Yasuko; Yano, Daisuke; Nagafusa, Kiyoshi; Kato, Takehide; Saito, Chieko; Uemura, Tomohiro; Ueda, Takashi; Nakano, Akihiko; Tasaka, Masao; Terao Morita, Miyo


    Plant vacuoles play critical roles in development, growth and stress responses. In mature cells, vacuolar membranes (VMs) display several types of structures, which are formed by invagination and folding of VMs into the lumenal side and can gradually move and change shape. Although such VM structures are observed in a broad range of tissue types and plant species, the molecular mechanism underlying their formation and maintenance remains unclear. Here, we report that a novel HEAT-repeat protein, SHOOT GRAVITROPISM6 (SGR6), of Arabidopsis is involved in the control of morphological changes and dynamics of VM structures in endodermal cells, which are the gravity-sensing cells in shoots. SGR6 is a membrane-associated protein that is mainly localized to the VM in stem endodermal cells. The sgr6 mutant stem exhibits a reduced gravitropic response. Higher plants utilize amyloplast sedimentation as a means to sense gravity direction. Amyloplasts are surrounded by VMs in Arabidopsis endodermal cells, and the flexible and dynamic structure of VMs is important for amyloplast sedimentation. We demonstrated that such dynamic features of VMs are gradually lost in sgr6 endodermal cells during a 30 min observation period. Histological analysis revealed that amyloplast sedimentation was impaired in sgr6. Detailed live-cell imaging analyses revealed that the VM structures in sgr6 had severe defects in morphological changes and dynamics. Our results suggest that SGR6 is a novel protein involved in the formation and/or maintenance of invaginated VM structures in gravity-sensing cells.

  9. Breeding for improved and novel starch characteristics in potato

    NARCIS (Netherlands)

    Nazarian, F.; Kok-Jacon, G.A.; Ji, Q.; Vincken, J.P.; Suurs, L.C.J.M.; Visser, R.G.F.


    Starch is the major form in which carbohydrates are stored and is present in almost all plant organs at one or another time during development. Storage starch is formed in amyloplasts as dense granules ranging in size from 1 to over 100 µ M. They are composed of an essential linear glucose polymer a

  10. In planta modification of potato starch granule biogenesis by different granule-bound fusion proteins

    NARCIS (Netherlands)

    Nazarian, F.


    Starch is composed of amylose and amylopectin and it is deposited in amyloplasts/choloroplasts as semi-crystalline granules. Many biosynthetic enzymes are involved in starch degradation and biosynthesis. Some microbial starch degrading enzymes have a Starch Binding Domain (SBD) which has affinity fo

  11. Influence of microgravity on root-cap regeneration and the structure of columella cells in Zea mays (United States)

    Moore, R.; McClelen, C. E.; Fondren, W. M.; Wang, C. L.


    We launched imbibed seeds and seedlings of Zea mays into outer space aboard the space shuttle Columbia to determine the influence of microgravity on 1) root-cap regeneration, and 2) the distribution of amyloplasts and endoplasmic reticulum (ER) in the putative statocytes (i.e., columella cells) of roots. Decapped roots grown on Earth completely regenerated their caps within 4.8 days after decapping, while those grown in microgravity did not regenerate caps. In Earth-grown seedlings, the ER was localized primarily along the periphery of columella cells, and amyloplasts sedimented in response to gravity to the lower sides of the cells. Seeds germinated on Earth and subsequently launched into outer space had a distribution of ER in columella cells similar to that of Earth-grown controls, but amyloplasts were distributed throughout the cells. Seeds germinated in outer space were characterized by the presence of spherical and ellipsoidal masses of ER and randomly distributed amyloplasts in their columella cells. These results indicate that 1) gravity is necessary for regeneration of the root cap, 2) columella cells can maintain their characteristic distribution of ER in microgravity only if they are exposed previously to gravity, and 3) gravity is necessary to distribute the ER in columella cells of this cultivar of Z. mays.

  12. Changes in the distribution of plastids and endoplasmic reticulum during cellular differentiation in root caps of Zea mays (United States)

    Moore, R.; McClelen, C. E.


    In calyptrogen cells of Zea mays, proplastids are distributed randomly throughout the cell, and the endoplasmic reticulum (ER) is distributed parallel to the cell walls. The differentiation of calyptrogen cells into columella statocytes is characterized by the following sequential events: (1) formation of ER complexes at the distal and proximal ends of the cell, (2) differentiation of proplastids into amyloplasts, (3) sedimentation of amyloplasts onto the distal ER complex, (4) breakdown of the distal ER complex and sedimentation of amyloplasts to the bottom of the cell, and (5) formation of sheets of ER parallel to the longitudinal cell walls. Columella statocytes located in the centre of the cap each possess 4530 +/- 780 micrometers2 of ER surface area, an increase of 670 per cent over that of calyptrogen cells. The differentiation of peripheral cells correlates positively with (1) the ER becoming arranged in concentric sheets, (2) amyloplasts and ER becoming randomly distributed, and (3) a 280 per cent increase in ER surface area over that of columella statocytes. These results are discussed relative to graviperception and mucilage secretion, which are functions of columella and peripheral cells, respectively.

  13. Ultrastructure of pea and cress root statocytes exposed to high gradient magnetic field (United States)

    Belyavskaya, N. A.; Chernishov, V. I.; Polishchuk, O. V.; Kondrachuk, A. V.

    As it was demonstrated by Kuznetsov & Hasenstein (1996) the high gradient magnetic field (HGMF) can produce a ponderomotive force that results in displacements of amyloplasts and causes the root response similar to the graviresponse. It was suggested that the HGMF could allow to imitate the effects of gravity in microgravity and/or change them in laboratory conditions correspondingly, as well as to study statolith-related processes in graviperception. Therefore, the correlation between the direction of the ponderomotive force resulting in statolith displacements and the direction of the HGMF-induced plant curvature can be the serious argument to support this suggestion and needs the detailed ultrastructural analysis. Seeds of dicotyledon Pisum sativum L. cv. Damir-2 and monocotyledon Lepidium sativum L. cv. P896 were soaked and grown in a vertical position on moist filter paper in chambers at room temperature. Tips of primary roots of vertical control, gravistimulated and exposed to HGMF seedlings were fixed for electron microscopy using conventional techniques. At ultrastructural level, we observed no significant changes in the volume of the individual statocytes or amyloplasts, relative volumes of cellular organelles (except vacuoles), number of amyloplasts per statocyte or surface area of endoplasmic reticulum. No consistent contacts between amyloplasts and any cellular structures, including plasma membrane, were revealed at any stage of magneto- and gravistimulation. By 5 min after onset of magnetostimulation, amyloplasts were located along cell wall distant from magnets. In HGMF, the locations of amyloplasts in columella cells were similar to those in horizontally-oriented roots up to 1 h stimulation. In the latter case, there were sometimes cytoplasmic spherical bodies with a dense vesicle-rich cytoplasm in pea statocytes, which were absent in seedlings exposed to HGMF. In cress root statocytes, both gravi- and magnetostimulation were found to cause the

  14. Transcriptome Analysis Suggests That Starch Synthesis May Proceed via Multiple Metabolic Routes in High Yielding Potato Cultivars

    DEFF Research Database (Denmark)

    Kaminski, Kacper Piotr; Petersen, Annabeth Høgh; Sønderkær, Mads;


    levels of transcripts for other enzymes involved in starch metabolism in comparison with medium and low yielding cultivars as determined by DeepSAGE transcriptome profiling. The decrease in PGM activity in Kuras was confirmed by measuring the enzyme activity from potato tuber extracts. Contrary......Background: Glucose-6-phosphate is imported into the amyloplast of potato tubers and thought to constitute the precursor for starch synthesis in potato tubers. However, recently it was shown that glucose-1-phosphate can also be imported into the amyloplast and incorporated into starch via an ATP...... independent mechanism under special conditions. Nonetheless, glucose-6-phosphate is believed to be the quantitatively important precursor for starch synthesis in potato. Principical finding: potato tubers of the high yielding cv Kuras had low gene expression of plastidial phophoglucomutase (PGM) and normal...

  15. Plastids and gravitropic sensing (United States)

    Sack, F. D.


    Data and theories about the identity of the mass that acts in gravitropic sensing are reviewed. Gravity sensing may have evolved several times in plants and algae in processes such as gravitropism of organs and tip-growing cells, gravimorphism, gravitaxis, and the regulation of cytoplasmic streaming in internodal cells of Chara. In the latter and in gravitaxis, the mass of the entire cell may function in sensing. But gravitropic sensing appears to rely upon the mass of amyloplasts that sediment since (i) the location of cells with sedimentation is highly regulated, (ii) such cells contain other morphological specializations favoring sedimentation, (iii) sedimentation always correlates with gravitropic competence in wild-type plants, (iv) magnetophoretic movement of rootcap amyloplasts mimics gravitropism, and (v) starchless and intermediate starch mutants show reduced gravitropic sensitivity. The simplest interpretation of these data is that gravitropic sensing is plastid-based.

  16. Conservation of the plastid sedimentation zone in all moss genera with known gravitropic protonemata (United States)

    Schwuchow, J. M.; Kern, V. D.; White, N. J.; Sack, F. D.


    Moss protonemata from several species are known to be gravitropic. The characterization of additional gravitropic species would be valuable to identify conserved traits that may relate to the mechanism of gravitropism. In this study, four new species were found to have gravitropic protonemata, Fissidens adianthoides, Fissidens cristatus, Physcomitrium pyriforme, and Barbula unguiculata. Comparison of upright and inverted apical cells of P. pyriforme and Fissidens species showed clear axial sedimentation. This sedimentation is highly regulated and not solely dependent on amyloplast size. Additionally, the protonemal tip cells of these species contained broad subapical zones that displayed lateral amyloplast sedimentation. The conservation of a zone of lateral sedimentation in a total of nine gravitropic moss species from five different orders supports the idea that this sedimentation serves a specialized and conserved function in gravitropism, probably in gravity sensing.

  17. Interaction between hydrotropism and gravitropism in seedling roots (United States)

    Kobayashi, A.; Takahashi, A.; Yamazaki, Y.; Kakimoto, Y.; Higashitani, A.; Fujii, N.; Takahashi, H.

    Roots display positive hydrotropism in response to a moisture gradient, which could play a role in avoiding drought stress. Because roots also respond to other stimuli such as gravity, touch and light and exhibit gravitropism, thigmotropism and phototropism, respectively, their growth orientation is determined by interaction among those tropisms. We have demonstrated the interaction between hydrotropism and gravitropism. For example, 1) agravitropic roots of pea mutant strongly respond to a moisture gradient and show positive hydrotropism by overcoming gravitropism, 2) in wild type pea roots hydrotropism is weak but pronounced when rotated on clinostat, 3) cucumber roots are positively gravitropic on the ground but become hydrotropic in microgravity, and 4) maize roots change their growth direction depending on the intensities of both gravistimulation and hydrostimulation. Here we found that Arabidopsis roots could display strong hydrotropism by overcoming gravitropism. It was discovered that amyloplasts in the columella cells are rapidly degraded upon exposure to a moisture gradient. Thus, degradation of amyloplasts could reduce the responsiveness to gravity, which could pronounce the hydrotropic response. In hydrotropically stimulated roots of pea seedlings, however, we could not observe a rapid degradation of amyloplasts in the columella cells. These results suggest that mechanism underlying the interaction between hydrotropism and gravitropism differs among plant species. To further study the molecular mechanism of hydrotropism and its interaction with gravitropism, we isolated unique mutants of Arabidopsis of which roots showed either ahydrotropism, reduced hydrotropism or negative hydrotropic response and examined their gravitropism, phototropism, waving response, amyloplast degradation and elongation growth. Based on the characterization of hydrotropic mutants, we will attempt to compare the mechanisms of the two tropisms and to clarify their cross talk for

  18. In planta modification of potato starch granule biogenesis by different granule-bound fusion proteins


    Nazarian, F.


    Starch is composed of amylose and amylopectin and it is deposited in amyloplasts/choloroplasts as semi-crystalline granules. Many biosynthetic enzymes are involved in starch degradation and biosynthesis. Some microbial starch degrading enzymes have a Starch Binding Domain (SBD) which has affinity for the starch granules on its own. In our laboratory, expression of SBD alone or fused to other effector proteins has been demonstrated. In industry, starch is modified after harvesting by chemical,...

  19. Kinetics of starch digestion and performance of broiler chickens


    Weurding, R.E.


    Keywords: starch, digestion rate, broiler chickens, peas, tapiocaStarch is stored in amyloplasts of various plants like cereals and legumes and seeds of these plants are used as feedstuffs for farm animals. Starch is the major energy source in broiler feeds. The properties of starch from different origin vary condiderably and these properties determine its resistance to enzymatic digestion. The objective of the research project described in this thesis was to study starch digestion behaviour ...

  20. Starch branching enzymes and their genes in Sorghum


    Mutisya, Joel


    Starch is an important raw material both for food and non-food purposes. It is synthesized and stored in source and sink tissues in plants. The starch deposited in amyloplasts of storage tissues possesses several physico-chemical properties, which makes it desirable for diverse applications. For industrial applications, starches high in either amylose or amylopectin are preferred in order to minimize chemical modification. The main purpose in our research is to understand starch synthesis in ...

  1. Gravitropic mechanisms derived from space experiments and magnetic gradients. (United States)

    Hasenstein, Karl H.; Park, Myoung Ryoul


    Gravitropism is the result of a complex sequence of events that begins with the movement of dense particles, typically starch-filled amyloplasts in response to reorientation. Although these organelles change positions, it is not clear whether the critical signal is derived from sedimentation or dynamic interactions of amyloplasts with relevant membranes. Substituting gravity by high-gradient magnetic fields (HGMF) provides a localized stimulus for diamagnetic starch that is specific for amyloplasts and comparable to gravity without affecting other organelles. Experiments with Brassica rapa showed induction of root curvature by HGMF when roots moved sufficiently close to the magnetic gradient-inducing foci. The focused and short-range effectiveness of HGMFs provided a gravity-like stimulus and affected related gene expression. Root curvature was sensitive to the mutual alignment between roots and HGMF direction. Unrelated to any HGMF effects, the size of amyloplasts in space-grown roots increased by 30% compared to ground controls and suggests enhanced sensitivity in a gravity-reduced environment. Accompanying gene transcription studies showed greater differences between HGMF-exposed and space controls than between space and ground controls. This observation may lead to the identification of gravitropism-relevant genes. However, space grown roots showed stronger transcription of common reference genes such as actin and ubiquitin in magnetic fields than in non-magnetic conditions. In contrast, α-amylase, glucokinase and PIN encoding genes were transcribed stronger under non-magnetic conditions than under HGMF. The large number of comparisons between space, ground, and HGMF prompted the assessment of transcription differences between root segments, root-shoot junction, and seeds. Because presumed transcription of reference genes varied more than genes of interest, changes in gene expression cannot be based on reference genes. The data provide an example of complex

  2. Inhibition of the gravitropic bending response of flowering shoots by salicylic acid. (United States)

    Friedman, Haya; Meir, Shimon; Halevy, Abraham H; Philosoph-Hadas, Sonia


    The upward gravitropic bending of cut snapdragon, lupinus and anemone flowering shoots was inhibited by salicylic acid (SA) applied at 0.5 mM and above. This effect was probably not due to acidification of the cytoplasm, since other weak acids did not inhibit bending of snapdragon shoots. In order to study its mode of inhibitory action, we have examined in cut snapdragon shoots the effect of SA on three processes of the gravity-signaling pathway, including: amyloplast sedimentation, formation of ethylene gradient across the stem, and differential growth response. The results show that 1 mM SA inhibited differential ethylene production rates across the horizontal stem and the gravity-induced growth, without significantly inhibiting vertical growth or amyloplast sedimentation following horizontal placement. However, 5 mM SA inhibited all three gravity-induced processes, as well as the growth of vertical shoots, while increasing flower wilting. It may, therefore, be concluded that SA inhibits bending of various cut flowering shoots in a concentration-dependent manner. Thus, at a low concentration SA exerts its effect in snapdragon shoots by inhibiting processes operating downstream to stimulus sensing exerted by amyloplast sedimentation. At a higher concentration SA inhibits bending probably by exerting general negative effects on various cellular processes.

  3. Cyclotron-based effects on plant gravitropism (United States)

    Kordyum, E.; Sobol, M.; Kalinina, Ia.; Bogatina, N.; Kondrachuk, A.

    Primary roots exhibit positive gravitropism and grow in the direction of the gravitational vector, while shoots respond negatively and grow opposite to the gravitational vector. We first demonstrated that the use of a weak combined magnetic field (CMF), which is comprised of a permanent magnetic field and an alternating magnetic field with the frequency resonance of the cyclotron frequency of calcium ions, can change root gravitropism from a positive direction to negative direction. Two-day-old cress seedlings were gravistimulated in a chamber that was placed into a μ-metal shield where this CMF was created. Using this "new model" of a root gravitropic response, we have studied some of its components including the movement of amyloplasts-statoliths in root cap statocytes and the distribution of Ca 2+ ions in the distal elongation zone during gravistimulation. Unlike results from the control, amyloplasts did not sediment in the distal part of a statocyte, and more Ca 2+ accumulation was observed in the upper side of a gravistimulated root for seedlings treated with the CMF. For plants treated with the CMF, it appears that a root gravitropic reaction occurs by a normal physiological process resulting in root bending although in the opposite direction. These results support the hypothesis that both the amyloplasts in the root cap statocytes and calcium are important signaling components in plant gravitropism.

  4. Gravisensing in roots (United States)

    Perbal, G.


    The mode of gravisensing in higher plants is not yet elucidated. Although, it is generally accepted that the amyloplasts (statoliths) in the root cap cells (statocytes) are responsible for susception of gravity. However, the hypothesis that the whole protoplast acts as gravisusceptor cannot be dismissed. The nature of the sensor that is able to transduce and amplify the mechanical energy into a biochemical factor is even more controversial. Several cell structures could potentially serve as gravireceptors: the endoplasmic reticulum, the actin network, the plasma membrane, or the cytoskeleton associated with this membrane. The nature of the gravisusceptors and gravisensors is discussed by taking into account the characteristics of the gravitropic reaction with respect to the presentation time, the threshold acceleration, the reciprocity rule, the deviation from the sine rule, the movement of the amyloplasts, the pre-inversion effect, the response of starch free and intermediate mutants and the effects of cytochalasin treatment. From this analysis, it can be concluded that both the amyloplasts and the protoplast could be the gravisusceptors, the former being more efficient than the latter since they can focus pressure on limited areas. The receptor should be located in the plasma membrane and could be a stretch-activated ion channel.

  5. Structural and Histochemical Characterization of Developing Rice Caryopsis

    Institute of Scientific and Technical Information of China (English)

    YU Xu-run; ZHOU Liang; XIONG Fei; WANG Zhong


    The development of pericarp, seed coat, starchy endosperm and aleurone of the rice caryopsis was investigated, histochemically and structurally, from the time of flowering to maturity. The results showed that during its growth, the maximum length of the caryopsis was attained first, followed by width and then thickness. Histochemical examination of the caryopsis showed that starch was mainly accumulated in the endosperm, but the endosperm showed no metabolic activity, while embryo and pericarp contained a few starch grains, and embryo and aleurone were strongly active. Aleuronic cells contained many aleurone grains and spherosomes, and aleurone in the dorsal region developed earlier and contained more layers of cells. Amyloplasts in endosperm contained many starch granules and were spherical at early stages but polyhedric at late stages. The protein bodies appeared later than amyloplasts, and the number of protein bodies in subaleurone was greater than those in the starchy endosperm. The white-belly portion of endosperm might be relative to the status of amyloplast development.

  6. Morphological Studies on the Mobilization of Reserves in Germinating Rice Seed : Decomposition process of starch granules


    ZAKARIA, Sabaruddin / MATSUDA, Toshiaki / NITTA, Youji


    The first step in the mobilization of the reserves in germinating rice seed was the decomposition of amylopllast envelopes in endosperm cells adjacent to scutellum.Destruction of amyloplast envelopes caused the separation of the starch granules.Small holes appeared on the surface of the released starch granule adjacent to scutellum at 3 days after imbibition(DAI).At later stages, the number and size of the holes increased on the surface of the starch granule.Ring-like structure in the interio...

  7. Fusion proteins comprising the catalytic domain of mutansucrase and a starch-binding domain can after the morphology of amylose-free potato starch granules during biosynthesis


    Nazarian, F.; Kok-Jacon, G.A.; Vincken, J.P.; Q. JI; Suurs, L.C.J.M.; Visser, R.G.F.


    It has been shown previously that mutan can be co-synthesized with starch when a truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The mutan seemed to adhere to the isolated starch granules, but it was not incorporated in the starch granules. In this study, GtfICAT was fused to the N- or C-terminus of a starch-binding domain (SBD). These constructs were introduced into two genetically different potato backgrounds (cv. Kardal and amf), in order to bring GtfICAT in more ...

  8. Induction of Plant Curvature by Magnetophoresis and Cytoskeletal Changes during Root Graviresponse (United States)

    Hasenstein, Karl H.; Kuznetsov, Oleg A.; Blancaflor, Eilson B.


    High gradient magnetic fields (HGMF) induce curvature in roots and shoots. It is considered that this response is likely to be based on the intracellular displacement of bulk starch (amyloplasts) by the ponderomotive force generated by the HGMF. This process is called magnetophoresis. The differential elongation during the curvature along the concave and convex flanks of growing organs may be linked to the microtubular and/or microfilament cytoskeleton. The possible existence of an effect of the HGMF on the cytoskeleton was tested for, but none was found. The application of cytoskeletal stabilizers or depolymerizers showed that neither microtubules, nor microfilaments, are involved in the graviresponse.

  9. Plastids and Carotenoid Accumulation. (United States)

    Li, Li; Yuan, Hui; Zeng, Yunliu; Xu, Qiang


    Plastids are ubiquitously present in plants and are the organelles for carotenoid biosynthesis and storage. Based on their morphology and function, plastids are classified into various types, i.e. proplastids, etioplasts, chloroplasts, amyloplasts, and chromoplasts. All plastids, except proplastids, can synthesize carotenoids. However, plastid types have a profound effect on carotenoid accumulation and stability. In this chapter, we discuss carotenoid biosynthesis and regulation in various plastids with a focus on carotenoids in chromoplasts. Plastid transition related to carotenoid biosynthesis and the different capacity of various plastids to sequester carotenoids and the associated effect on carotenoid stability are described in light of carotenoid accumulation in plants. PMID:27485226

  10. Immunoelectron microscopic localization of calmodulin in corn root cells

    Institute of Scientific and Technical Information of China (English)



    Methods for the localization of plant calmodulin by immuno-gold and immuno-peroxidase electron microscopy have been developed. In both corn root-cap cells and meristematic cells, calmodulin was found to be localized in the nucleus, cytoplasm, mitochondria as well as in the cell wall, In the meristematic cells, calmodulin was distinctly localized on the plasma membrane, cytoplasmic face of rough endoplasmic rcticulum and polyribosomes. Characteristically, calmodulin was present in the amyloplasts of root-cap cells. The widespread distribution of calmodulin may reflect its plciotropic functions in plant cellular activities.

  11. Transcriptome analysis suggests that starch synthesis may proceed via multiple metabolic routes in high yielding potato cultivars.

    Directory of Open Access Journals (Sweden)

    Kacper Piotr Kaminski

    Full Text Available BACKGROUND: Glucose-6-phosphate is imported into the amyloplast of potato tubers and thought to constitute the precursor for starch synthesis in potato tubers. However, recently it was shown that glucose-1-phosphate can also be imported into the amyloplast and incorporated into starch via an ATP independent mechanism under special conditions. Nonetheless, glucose-6-phosphate is believed to be the quantitatively important precursor for starch synthesis in potato. PRINCIPAL FINDING: Potato tubers of the high yielding cv Kuras had low gene expression of plastidial phophoglucomutase (PGM and normal levels of transcripts for other enzymes involved in starch metabolism in comparison with medium and low yielding cultivars as determined by DeepSAGE transcriptome profiling. The decrease in PGM activity in Kuras was confirmed by measuring the enzyme activity from potato tuber extracts. Contrary to expectations, this combination lead to a higher level of intracellular glucose-1-phosphate (G1P in Kuras suggesting that G1P is directly imported into plastids and can be quantitatively important for starch synthesis under normal conditions in high yielding cultivars. SIGNIFICANCE: This could open entirely new possibilities for metabolic engineering of the starch metabolism in potato via the so far uncharacterized G1P transporter. The perspectives are to increase yield and space efficiency of this important crop. In the light of the increasing demands imposed on agriculture to support a growing global population this presents an exciting new possibility.

  12. Effects of inversion on plastid position and gravitropism in Ceratodon protonemata (United States)

    Schwuchow, J.; Sack, F. D.


    When dark-grown tip cells of protonemata of the moss Ceratodon purpureus are turned to the horizontal, plastids first sediment towards gravity in a specific zone and then the tip curves upward. To determine whether gravitropism and plastid sedimentation occur in other orientations, protonemata were reoriented to angles other than 90 degrees. Qualitative and quantitative light microscopic observations show that plastid sedimentation along the cell axis occurs in both upright and inverted cells. However, only some plastids fall and sedimentation is incomplete; plastids remain distributed throughout the length of the cell, and those plastids that sediment do not fall all the way to the bottom of the cell. Tip cells are gravitropic regardless of stimulation angle, and as in higher plants, the maximal rate of initial curvature is in response to a 120 degrees reorientation. Infrared videomicroscopy, time-lapse studies of living, inverted protonemata indicate that amyloplast sedimentation precedes upward curvature. Together, these data further support (i) the hypothesis that amyloplast sedimentation functions in gravitropic sensing in these cells, and (ii) the idea that gravity affected the evolution of cell organization.

  13. Plastid position in Arabidopsis columella cells is similar in microgravity and on a random-positioning machine (United States)

    Kraft, T. F.; van Loon, J. J.; Kiss, J. Z.


    In order to study gravity effects on plant structure and function, it may become necessary to remove the g-stimulus. On Earth, various instruments such as clinostats have been used by biologists in an attempt to neutralize the effects of gravity. In this study, the position of amyloplasts was assayed in columella cells in the roots of Arabidopsis thaliana (L.) Heynh. seedlings grown in the following conditions: on Earth, on a two-dimensional clinostat at 1 rpm, on a three-dimensional clinostat (also called a random-positioning machine, or an RPM), and in space (true microgravity). In addition, the effects of these gravity treatments on columella cell area and plastid area also were measured. In terms of the parameters measured, only amyloplast position was affected by the gravity treatments. Plastid position was not significantly different between spaceflight and RPM conditions but was significantly different between spaceflight and the classical two-dimensional clinostat treatments. Flanking columella cells showed a greater susceptibility to changes in gravity compared to the central columella cells. In addition, columella cells of seedlings that were grown on the RPM did not exhibit deleterious effects in terms of their ultrastructure as has been reported previously for seedlings grown on a two-dimensional clinostat. This study supports the hypothesis that the RPM provides a useful simulation of weightlessness.

  14. Phototropism and gravitropism in lateral roots of Arabidopsis. (United States)

    Kiss, John Z; Miller, Kelley M; Ogden, Lisa A; Roth, Kelly K


    Gravitropism and, to a lesser extent, phototropism have been characterized in primary roots, but little is known about structural/functional aspects of these tropisms in lateral roots. Therefore, in this study, we report on tropistic responses in lateral roots of Arabidopsis thaliana. Lateral roots initially are plagiogravitropic, but when they reach a length of approximately 10 mm, these roots grow downward and exhibit positive orthogravitropism. Light and electron microscopic studies demonstrate a correlation between positive gravitropism and development of columella cells with large, sedimented amyloplasts in wild-type plants. Lateral roots display negative phototropism in response to white and blue light and positive phototropism in response to red light. As is the case with primary roots, the photoresponse is weak relative to the graviresponse, but phototropism is readily apparent in starchless mutant plants, which are impaired in gravitropism. To our knowledge, this is the first report of phototropism of lateral roots in any plant species.

  15. Gravitropism in leaves of Arabidopsis thaliana (L.) Heynh. (United States)

    Mano, Eriko; Horiguchi, Gorou; Tsukaya, Hirokazu


    In higher plants, stems and roots show negative and positive gravitropism, respectively. However, current knowledge on the graviresponse of leaves is lacking. In this study, we analyzed the positioning and movement of rosette leaves of Arabidopsis thaliana under light and dark conditions. We found that the radial positioning of rosette leaves was not affected by the direction of gravity under continuous white light. In contrast, when plants were shifted to darkness, the leaves moved upwards, suggesting negative gravitropism. Analysis of the phosphoglucomutase and shoot gravitropism 2-1 mutants revealed that the sedimenting amyloplasts in the leaf petiole are important for gravity perception, as is the case in stems and roots. In addition, our detailed physiological analyses revealed a unique feature of leaf movement after the shift to darkness, i.e. movement could be divided into negative gravitropism and nastic movement. The orientation of rosette leaves is ascribed to a combination of these movements.

  16. [Enhancement of photoassimilate utilization by manipulation of the ADPglucose pyrophosphorylase gene]. Progress report, [March 15, 1989--April 14, 1990

    Energy Technology Data Exchange (ETDEWEB)

    Okita, T.W.


    The long term aim of this project is to assess the feasibility of increasing the conversion of photosynthate into starch via manipulation of the gene that encodes for ADPglucose pyrophosphorylase, a key regulatory enzyme of starch biosynthesis. In developing storage tissues such as cereal seeds and tubers, starch biosynthesis is regulated by the gene activation and expression of ADPglucose pyrophosphorylase, starch synthase, branching enzyme and other ancillary starch modifying enzymes, as well as the allosteric-controlled behavior of ADPglucose pyrophosphorylase activity. During the last two years we have obtained information on the structure of this enzyme from both potato tuber and rice endosperm, using a combination of biochemical and molecular biological approaches. Moreover, we present evidence that this enzyme may be localized at discrete regions of the starch grain within the amyloplast, and plays a role in controlling overall starch biosynthesis in potato tubers.

  17. Distribution of calmodulin in corn seedlings - Immunocytochemical localization in coleoptiles and root apices (United States)

    Dauwalder, M.; Roux, S. J.


    Immunofluorescence techniques have been used to study the distribution of calmodulin in several tissues in etiolated corn (Zea mays, var. Bear Hybrid) seedlings. Uniform staining was seen in the background cytoplasm of most cell types. Cell walls and vacuoles were not stained. In coleoptile mesophyll cells the nucleoplasm of most nuclei was stained as was the stroma of most amyloplasts. The lumen border of mature tracheary elements in coleoptiles also stained. In the rootcap the most intensely stained regions were the cytoplasms of columella cells and of the outermost cells enmeshed in the layer of secreted slime. Nuclei in the rootcap cells did not stain distinctly, but those in all cell types of the root meristem did. Also in the root meristem, the cytoplasm of metaxylem elements stained brightly. These results are compared and contrasted with previous data on the localization of calmodulin in pea root apices and epicotyls and discussed in relation to current hypotheses on mechanisms of gravitropism.

  18. Molecular mechanisms of gravity perception and signal transduction in plants. (United States)

    Kolesnikov, Yaroslav S; Kretynin, Serhiy V; Volotovsky, Igor D; Kordyum, Elizabeth L; Ruelland, Eric; Kravets, Volodymyr S


    Gravity is one of the environmental cues that direct plant growth and development. Recent investigations of different gravity signalling pathways have added complexity to how we think gravity is perceived. Particular cells within specific organs or tissues perceive gravity stimulus. Many downstream signalling events transmit the perceived information into subcellular, biochemical, and genomic responses. They are rapid, non-genomic, regulatory, and cell-specific. The chain of events may pass by signalling lipids, the cytoskeleton, intracellular calcium levels, protein phosphorylation-dependent pathways, proteome changes, membrane transport, vacuolar biogenesis mechanisms, or nuclear events. These events culminate in changes in gene expression and auxin lateral redistribution in gravity response sites. The possible integration of these signalling events with amyloplast movements or with other perception mechanisms is discussed. Further investigation is needed to understand how plants coordinate mechanisms and signals to sense this important physical factor. PMID:26215561

  19. The Quest for Golden Bananas: Investigating Carotenoid Regulation in a Fe'i Group Musa Cultivar. (United States)

    Buah, Stephen; Mlalazi, Bulukani; Khanna, Harjeet; Dale, James L; Mortimer, Cara L


    The regulation of carotenoid biosynthesis in a high-carotenoid-accumulating Fe'i group Musa cultivar, "Asupina", has been examined and compared to that of a low-carotenoid-accumulating cultivar, "Cavendish", to understand the molecular basis underlying carotenogenesis during banana fruit development. Comparisons in the accumulation of carotenoid species, expression of isoprenoid genes, and product sequestration are reported. Key differences between the cultivars include greater carotenoid cleavage dioxygenase 4 (CCD4) expression in "Cavendish" and the conversion of amyloplasts to chromoplasts during fruit ripening in "Asupina". Chromoplast development coincided with a reduction in dry matter content and fruit firmness. Chromoplasts were not observed in "Cavendish" fruits. Such information should provide important insights for future developments in the biofortification and breeding of banana.

  20. A new genetic factor for root gravitropism in rice (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    Jianghua SHI; Xi HAO; Zhong-chang WU; Ping WU


    Root gravitropism is one of the important factors to determine root architecture. To understand the mechanism un-derlying root gravitropism, we isolated a rice (Xiushui63) mutant defective in root gravitropism, designated as glsl. Vertical sections of root caps revealed that glsl mutant displayed normal distribution of amyloplast in the columella cells compared with the wild type. The glsl mutant was less sensitive to 2,4-dichlorophenoxyacetic acid (2,4-D) and a-naphthaleneacetic acid (NAA) than the wild type. Genetic analysis indicated that the phenotype of glsl mutant was caused by a single recessive mutation, which is mapped in a 255-kb region between RM16253 and CAPSI on the short arm of chromosome 4.

  1. Mechanism of dynamic reorientation of cortical microtubules due to mechanical stress

    CERN Document Server

    Muratov, Alexander


    Directional growth caused by gravitropism and corresponding bending of plant cells has been explored since 19th century, however, many aspects of mechanisms underlying the perception of gravity at the molecular level are still not well known. Perception of gravity in root and shoot gravitropisms is usually attributed to gravisensitive cells, called statocytes, which exploit sedimentation of macroscopic and heavy organelles, amyloplasts, to sense the direction of gravity. Gravity stimulus is then transduced into distal elongation zone, which is several mm far from statocytes, where it causes stretching. It is suggested that gravity stimulus is conveyed by gradients in auxin flux. We propose a theoretical model that may explain how concentration gradients and/or stretching may indirectly affect the global orientation of cortical microtubules, attached to the cell membrane and induce their dynamic reorientation perpendicular to the gradients. In turn, oriented microtubules arrays direct the growth and orientatio...

  2. Genetic analysis of gravity signal transduction in roots (United States)

    Masson, Patrick; Strohm, Allison; Baldwin, Katherine

    To grow downward into the soil, roots use gravity as a guide. Specialized cells, named stato-cytes, enable this directional growth response by perceiving gravity. Located in the columella region of the cap, these cells sense a reorientation of the root within the gravity field through the sedimentation of, and/or tension/pressure exerted by, dense amyloplasts. This process trig-gers a gravity signal transduction pathway that leads to a fast alkalinization of the cytoplasm and a change in the distribution of the plasma membrane-associated auxin-efflux carrier PIN3. The latter protein is uniformly distributed within the plasma membrane on all sides of the cell in vertically oriented roots. However, it quickly accumulates at the bottom side upon gravis-timulation. This process correlates with a preferential transport of auxin to the bottom side of the root cap, resulting in a lateral gradient across the tip. This gradient is then transported to the elongation zone where it promotes differential cellular elongation, resulting in downward curvature. We isolated mutations that affect gravity signal transduction at a step that pre-cedes cytoplasmic alkalinization and/or PIN3 relocalization and lateral auxin transport across the cap. arg1 and arl2 mutations identify a common genetic pathway that is needed for all three gravity-induced processes in the cap statocytes, indicating these genes function early in the pathway. On the other hand, adk1 affects gravity-induced PIN3 relocalization and lateral auxin transport, but it does not interfere with cytoplasmic alkalinization. ARG1 and ARL2 encode J-domain proteins that are associated with membranes of the vesicular trafficking path-way whereas ADK1 encodes adenosine kinase, an enzyme that converts adenosine derived from nucleic acid metabolism and the AdoMet cycle into AMP, thereby alleviating feedback inhibi-tion of this important methyl-donor cycle. Because mutations in ARG1 (and ARL2) do not completely eliminate

  3. Genetic Analysis of Gravity Signal Transduction in Arabidopsis Roots (United States)

    Masson, Patrick; Strohm, Allison; Barker, Richard; Su, Shih-Heng

    Like most other plant organs, roots use gravity as a directional guide for growth. Specialized cells within the columella region of the root cap (the statocytes) sense the direction of gravity through the sedimentation of starch-filled plastids (amyloplasts). Amyloplast movement and/or pressure on sensitive membranes triggers a gravity signal transduction pathway within these cells, which leads to a fast transcytotic relocalization of plasma-membrane associated auxin-efflux carrier proteins of the PIN family (PIN3 and PIN7) toward the bottom membrane. This leads to a polar transport of auxin toward the bottom flank of the cap. The resulting lateral auxin gradient is then transmitted toward the elongation zones where it triggers a curvature that ultimately leads to a restoration of vertical downward growth. Our laboratory is using strategies derived from genetics and systems biology to elucidate the molecular mechanisms that modulate gravity sensing and signal transduction in the columella cells of the root cap. Our previous research uncovered two J-domain-containing proteins, ARG1 and ARL2, as contributing to this process. Mutations in the corresponding paralogous genes led to alterations of root and hypocotyl gravitropism accompanied by an inability for the statocytes to develop a cytoplasmic alkalinization, relocalize PIN3, and transport auxin laterally, in response to gravistimulation. Both proteins are associated peripherally to membranes belonging to various compartments of the vesicular trafficking pathway, potentially modulating the trafficking of defined proteins between plasma membrane and endosomes. MAR1 and MAR2, on the other end, are distinct proteins of the plastidic outer envelope protein import TOC complex (the transmembrane channel TOC75 and the receptor TOC132, respectively). Mutations in the corresponding genes enhance the gravitropic defects of arg1. Using transformation-rescue experiments with truncated versions of TOC132 (MAR2), we have shown

  4. Statolith action by the numbers: Physics and feasbility (United States)

    Todd, Paul; Yoder, Thomas; Staehelin, L. Andrew


    All modern (and ancient) studies make it clear that statolith motion is required for gravisensing, and some evidence indicates a role for statolith-microfilament interaction. In this study two components of statolith action are considered quantitatively: (1) the movement of statoliths through the columella cell cytoplasm and (2) forces at the site of action of the statolith. (1) Statoliths move through the cytoplasm in the presence of viscous and elastic forces that may be considered separately. The viscous component may be characterized as a solution with a viscosity of approximately 40 centiPoise. Statoliths are deflected from perfectly vertical trajectories by various obstacles, including actin filaments, and their velocities are influenced by interactions between statoliths. Channeling flow is commonly observed, but this cannot be due to the breaking of actin filaments by the force of the sedimenting statolith, as about 600 pN force is required to break a filament, and the force due to gravity on the amyloplast is about 0.07 pN. Under least viscous conditions the randomly-directed Brownian diffusion velocity of the amyloplast is about 10% the sedimentation velocity. (2) Intimate association of statoliths with the cortical ER region requiring gravitational force is postulated. Thin-section micrographs clearly show an interface between the cortex and the central cytoplasm of the columella cell in vascular plant species that have been studied. Whether or not an exchange of chemical components is required, experiments, including observations in low gravity, have demonstrated that the statolith is shallowly embedded in this cortical region to the extent that deformation occurs consistent with Newton's third law. Relieving the gravitational force results in instantaneous motion of the amyloplast away from the cortex. The following inquiry was made concerning the physics at the interface between the statolith and the material of the cortical ER region. How much force

  5. Gravitropism of cut shoots is mediated by oxidative processes: A physiological and molecular study (United States)

    Philosoph-Hadas, Sonia; Friedman, Haya; Meir, Shimon


    The signal transduction events occurring during shoot gravitropism are mediated through amyloplasts sedimentation, reorientation of actin filaments in the endodermis, and differential changes in level and action of auxin, associated with differential growth leading to shoot curvature. Since increase in reactive oxygen species (ROS) was shown to be associated with growth, we examined the possible use of antioxidants in controlling the gravitropic response, via their interaction with events preceding shoot bending. Reoriented snapdragon (Antirrhinum majus L.) spikes and tomato (Solanum lycopersicum cv. MicroTom) shoots showed a visual upward bending after a lag period of 3 or 5 h, respectively, which was inhibited by the antioxidants N-acetyl-cysteine (NAC) and reduced glutathione (GSH). This suggests the involvement of oxidative reactions in the process. The two antioxidants prevented the sedimentation of amyloplasts to the bottom of the endodermis cells following 0.5-5 h of snapdragon shoot reorientation, suggesting that oxidative reactions are involved already at a very early signal perception stage prior to the visual bending. In addition, a differential distribution in favor of the lower shoot side of various oxidative elements, including H2O2 concentrations and activity of the NADPH-oxidase enzyme, was observed during reorientation of snapdragon spikes. Application of the two antioxidants reduced the levels of these elements and abolished their differential distribution across the shoot. On the other hand, the activity of the antioxidative enzyme, superoxide dismutase (SOD), which was not differentially distributed across the shoot, increased significantly following application of the two antioxidants. The auxin redistribution in reoriented shoots was analyzed using transgenic tomato plants expressing the GUS reporter gene under the Aux/IAA4 promoter (a generous gift of M. Bouzayen, France). GUS response, detected in control shoots 4 h after their reorientation

  6. Comparison of glycerolipid biosynthesis in non-green plastids from sycamore (Acer pseudoplatanus) cells and cauliflower (Brassica oleracea) buds. (United States)

    Alban, C; Joyard, J; Douce, R


    The availability of methods to fractionate non-green plastids and to prepare their limiting envelope membranes [Alban, Joyard & Douce (1988) Plant Physiol. 88, 709-717] allowed a detailed analysis of the biosynthesis of lysophosphatidic acid, phosphatidic acid, diacylglycerol and monogalactosyl-diacylglycerol (MGDG) in two different types of non-green starch-containing plastids: plastids isolated from cauliflower buds and amyloplasts isolated from sycamore cells. An enzyme [acyl-ACP (acyl carrier protein):sn-glycerol 3-phosphate acyltransferase) recovered in the soluble fraction of non-green plastids transfers oleic acid from oleoyl-ACP to the sn-1 position of sn-glycerol 3-phosphate to form lysophosphatidic acid. Then a membrane-bound enzyme (acyl-ACP:monoacyl-sn-glycerol 3-phosphate acyltransferase), localized in the envelope membrane, catalyses the acylation of the available sn-2 position of 1-oleoyl-sn-glycerol 3-phosphate by palmitic acid from palmitoyl-ACP. Therefore both the soluble phase and the envelope membranes are necessary for acylation of sn-glycerol 3-phosphate. The major difference between cauliflower (Brassica oleracea) and sycamore (Acer pseudoplatanus) membranes is the very low level of phosphatidate phosphatase activity in sycamore envelope membrane. Therefore, very little diacylglycerol is available for MGDG synthesis in sycamore, compared with cauliflower. These findings are consistent with the similarities and differences described in lipid metabolism of mature chloroplasts from 'C18:3' and 'C16:3' plants (those with MGDG containing C18:3 and C16:3 fatty acids). Sycamore contains only C18 fatty acids in MGDG, and the envelope membranes from sycamore amyloplasts have a low phosphatidate phosphatase activity and therefore the enzymes of the Kornberg-Pricer pathway have a low efficiency of incorporation of sn-glycerol 3-phosphate into MGDG. By contrast, cauliflower contains MGDG with C16:3 fatty acid, and the incorporation of sn-glycerol 3

  7. Tropism in azalea and lily flowers (United States)

    Shimizu, M.; Tomita-Yokotani, K.; Nakamura, T.; Yamashita, M.

    Flowers have coevolved with pollinator animals. Some flowers have the up-down directional features in their form and orientation, which results the higher success of pollination under the influence of gravity. Azalea, Rhododendron pulchrum, flower responds against gravity, and orients the specific petal at its top. This petal with honey mark guides pollinator animals to nectary of the flower. Pistil and stamen bend upward by sensing gravity, and increase probability of their contact with pollinator. There was large sediment amyloplast found in sectioned tissue of style. In addition to this action of gravity, phototropic response was also observed at lesser degree, while the gravitational cue was removed by the 3D-clinorotation of the plant. In contrast to azalea, pistil of lily flower senses light in order to determine the direction of bending. Lily, Lilium cv. 'Casablanca', tepals open horizontally or slightly inclined downward. After its anthesis, pistil and stamen start to bend upward by light. Gravity induced no tropic response at all, evidenced by the experiment conducted under dark. Sediment amyloplast was not found in lily style. Phototropic response of pistil and stamen in lily was activated by blue light even at lower energy density. On the other hand, red light was not effective to induce the tropic response even with substantial energy density. This action spectrum of light agreed with those for the phototropism shown in coleoptile of monocotyledonous plants. Because the tropism of style was not hindered at removal of stigma, reception site for incident light is neither restricted to stigma nor its close vicinity, but distributes through style. The process of lily pistil elongation was analyzed in details to identify the site of its initiation and propagation of bending movement through the anthesis period. Elongation started at basal part of pistil and propagated towards its top after opening of perianth. Steep bending occurred at the basal zone of

  8. Graviperception of lentil seedling roots grown in space (Spacelab D1 Mission). (United States)

    Perbal, G; Driss-Ecole, D; Rutin, J; Salle, G


    The growth and graviresponsiveness of roots were investigated in lentil seedlings (Lens culinaris L. cv. Verte du Puy) grown (1) in microgravity, (2) on a 1 g centrifuge in space, (3) in microgravity and then placed on the 1 g centrifuge for 3 h, (4) on the ground. Dry seeds were hydrated in space (except for the ground control) and incubated for 25 h at 22 degrees C in darkness. At the end of the experiment, the seedlings were photographed and fixed in glutaraldehyde in a Biorack glove box. Root length was similar for seedlings grown in space and for the ground and the 1 g centrifuge controls. The direction of root growth in the microgravity sample deviated strongly from the initial orientation of the roots of the dry seeds. This deviation could be due to spontaneous curvatures similar to those observed on clinostats. When lentil seedlings were first grown in microgravity for 25 h and then placed on the 1 g centrifuge for 3 h, their roots bent strongly under the effect of the centrifugal acceleration. The amplitude of root curvature on the centrifuge was not significantly different from that observed on ground controls growing in the vertical position and placed in the horizontal position for 3 h. The gravisensitivity of statocytes differentiated in microgravity was similar to that of statocytes differentiated on earth. There were no qualitative differences in the ultrastructural features of the gravisensing cells in microgravity and in the 1 g centrifuge and ground controls. However, the distribution of statoliths in the gravisensing cells was different in microgravity: most of them were observed in the proximal part of these cells. Thus, these organelles were not distributed at random, which is in contradiction with results obtained with clinostats. The distal complex of endoplasmic reticulum in the statocytes was not in contact with the amyloplasts. Contact and pressure of amyloplasts on the tubules were not prerequisites for gravisensing. The results obtained

  9. Changes in root gravitropism, ultrastructure, and calcium balance of pea root statocytes induced by A23187 (United States)

    Belyavskaya, N.

    The role for calcium in the regulation of a wide variety of cellular events in plants is well known. Calcium signaling has been implicated in plant gravitropism. A carboxylic acid antibiotic A23187 (calcimycin) has been widely used in biological studies since it can translocate calcium across membranes. Seedlings of Pisum sativum L. cv. Uladovsky germinated in a vertically oriented cylinder of moist filter paper soaked in water during 4.5 day had been treated with 10-5 M A23187 for 12 hr. Tips of primary roots of control and A23187-treated pea seedlings were fixed for electron microscopy and electron cytochemistry. Experiments with Pisum sativum 5- day seedlings placed horizontally for 4 h after treatment with 10 μM A23187 during 12 h found that the graviresponsiveness of their primary roots was lost completely (91 % of roots) or inhibited (24 +/- 6° in comparison with 88 +/- 8° in control). At ultrastructural level, there were observed distribution of amyloplasts around the nucleus, remarkable lengthening of statocytes, advanced vacuolization, changes in dictyosome structure, ER fragmentation, cell wall thinning in A23187-treated statocytes. Cytochemical study has indicated that statocytes exposed to calcimycin have contained a number of Ca-pyroantimonate granules detected Ca 2 + ions in organelles and hyaloplasm (unlike the control ones). The deposits were mainly associated with the plasma membrane. Among organelles, mitochondria were notable for their ability to accumulate Ca 2 +. In amyloplasts, a fine precipitate was predominately located in their stroma and envelope lumens. In cell walls, deposits of the reaction product were observed along the periphery and in the median zone. Localization of electron-dense granules of lead phosphate, which indicated Ca 2 +- ATPase activities in pea statocytes exposed to A23187, was generally consistent with that in untreated roots. Apart from plasma membrane, chromatin, and nucleolus components, the cytochemical reaction

  10. Displacement of organelles in plant gravireceptor cells by vibrational forces and ultrasound. (United States)

    Kuznetsov, O.; Nechitailo, G.; Kuznetsov, A.

    Plant gravity perception can be studied by displacing statoliths inside receptor cells by forces other than gravity. Due to mechanical heterogeneity of statocytes various ponderomotive forces can be used for this purpose. In a plant subjected to non- symmetric vibrations statoliths experience inertial force proportional to the difference between their density and that of cytoplasm and to the instantaneous acceleration of the cell. This force causes cyclic motion of statoliths relative to cytoplasm and, depending on the profile of oscillations, can result in a net displacement of them (due to complex rheology of the cell interior), similar to sedimentation. This can be described as "vibrational" ponderomotive force acting on the statoliths. Vertically growing Arabidopsis seedlings, subjected to horizontal, sawtooth shaped oscillations (250 Hz, 1.5 mm amplitude), showed 17+/-2o root curvature toward and shoot curvature of 11+/-3o against the stronger acceleration. When the polarity of the oscillations was reversed, the direction of curvature of shoots and roots was also reversed. Control experiments with starchless mutants (TC7) produced no net curvature, which indicates that dense starch-filled amyloplasts are needed for the effect. These control experiments also eliminate touch-induced reactions or other side-effects as the cause of the curvature. Linum roots curved 25+/-7o . Ceratodon protonemata subjected to the same oscillations have shown displacement of plastids and curvature consistent with the pattern observed during graviresponse: positively gravitropic wwr mutant curved in the direction of the plastid displacement, WT curved in the opposite direction. Acoustic ponderomotive forces, originating from transfer of a sonic beam momentum to the medium due to sound scattering and attenuation in a mechanically heterogeneous system, also can displace statoliths. Vertical flax seedlings curved away from the ultrasonic source (800 kHz, 0.1 W/cm2 ) presumably as a

  11. Columella cells revisited: novel structures, novel properties, and a novel gravisensing model (United States)

    Staehelin, L. A.; Zheng, H. Q.; Yoder, T. L.; Smith, J. D.; Todd, P.


    A hundred years of research has not produced a clear understanding of the mechanism that transduces the energy associated with the sedimentation of starch-filled amyloplast statoliths in root cap columella cells into a growth response. Most models postulate that the statoliths interact with microfilaments (MF) to transmit signals to the plasma membrane (or ER), or that sedimentation onto these organelles produces the signals. However, no direct evidence for statolith-MF links has been reported, and no asymmetric structures of columella cells have been identified that might explain how a root turned by 90 degrees knows which side is up. To address these and other questions, we have (1) quantitatively examined the effects of microgravity on the size, number, and spatial distribution of statoliths; (2) re-evaluated the ultrastructure of columella cells in high-pressure frozen/freeze-substituted roots; and (3) followed the sedimentation dynamics of statolith movements in reoriented root tips. The findings have led to the formulation of a new model for the gravity-sensing apparatus of roots, which envisages the cytoplasm pervaded by an actin-based cytoskeletal network. This network is denser in the ER-devoid central region of the cell than in the ER-rich cell cortex and is coupled to receptors in the plasma membrane. Statolith sedimentation is postulated to disrupt the network and its links to receptors in some regions of the cell cortex, while allowing them to reform in other regions and thereby produce a directional signal.

  12. A weak combined magnetic field changes root gravitropism (United States)

    Kordyum, E. L.; Bogatina, N. I.; Kalinina, Ya. M.; Sheykina, N. V.

    Although gravitropism has been studied for many decades, many questions on plant gravitropism, including the participation of Ca 2+ ions in graviperception and signal transduction, remain open and require new experiments. We have studied gravistimulation and root gravitropism in the presence of the weak, alternating magnetic field that consisted of a sinusoidal frequency of 32 Hz inside a μ-metal shield. We discovered that this field changes normally positively gravitropic cress root to exhibit negative gravitropism. Because the combined magnetic field was adjusted to the cyclotron frequency of Ca 2+ ions, the obtained data suggest that calcium ion participate in root gravitropism. Simultaneous application of the oscillating magnetic field of the same frequency ion induce oscillation of Ca 2+ ions and can change the rate and/or the direction of Ca 2+ ion flux in roots. Control and magnetic field-exposed roots were examined for change in the distribution of amyloplasts and cellular organelles by light, electron, and confocal laser microscopy.

  13. The RHG gene is involved in root and hypocotyl gravitropism in Arabidopsis thaliana. (United States)

    Fukaki, H; Fujisawa, H; Tasaka, M


    In higher plants, shoots show negative gravitropism and roots show positive gravitropism. To elucidate the molecular mechanisms of root and hypocotyl gravitropism, we segregated the second mutation from the original phyB-1 mutant line which impaired both root and hypocotyl gravitropism and characterized this novel mutation named rhg (for root and hypocotyl gravitropism). The rhg is a single recessive nuclear mutation and it is mapped on the lower part of the chromosome 1. Analyses on the gravitropic responses of the rhg mutant indicate that root and hypocotyl gravitropism are severely impaired but inflorescence stem gravitropism is not affected by the rhg mutation. In the rhg mutant seedlings, amyloplasts (statoliths for gravity-perception) were present in the presumptive statocytes of roots and hypocotyls. Phototropism by roots and hypocotyls was not impaired in the rhg mutant. These results suggest that the RHG gene product probably acts on the gravity-perception and/or the gravity-signal transduction in root and hypocotyl gravitropism. This is the first report about the genetic locus specifically involved in both root and hypocotyl gravitropism but not inflorescence stem gravitropism, supporting our hypothesis that the mechanisms of gravitropism are genetically different between hypocotyls and inflorescence stems.

  14. Rape embryogenesis. IV. Appearance and disappearance of starch during embryo development

    Directory of Open Access Journals (Sweden)

    Teresa Tykarska


    Full Text Available Starch appears first in the suspensor of the proembryo with two-cell apical part. It is observed in the embryo proper from the octant stage. At first it is visible in all the embryo cells in the form of minute transient grains which disappear during cell divisions. But the columella mother cells and their derivatives have persistent large grains. When the embryo turns green in the heart stage a gradual accumulation of storage starch begins and lasts to the end of embryogenesis. Storage starch grains appear first in the auter cortex layers of the hypocotyl where the largest grains are to be found later, and afterwards in all the other tissues. Starch is usually absent in the frequently dividing cells, but even there it appears in the form of minute grains after the end of cell divisions. Disappearance of starch starts when the intensive green colour of the seed coat begins to fade. The first to disappear are the smallest granules in the regions where they were noted latest. In the embryo axis the starch grains remain deposited longest in dermatogen and cortex cells in the lower hypocotyl part. They are visible there, still when the seed turns brown. In black seeds starch may be only found in the columella the cells of which throughout embryogenesis contain amyloplasts filled with starch. These grains disappear completely at the time when the seeds become dry.

  15. Is the availability of substrate for the tricarboxylic acid cycle a limiting factor for uncoupled respiration in sycamore (Acer pseudoplatanus) cells? (United States)

    Journet, E P; Bligny, R; Douce, R


    Protoplasts obtained from sycamore (Acer pseudoplatanus) cell suspensions were found to be highly intact and to retain a high rate of O2 consumption. If the protoplasts were taken up and expelled through a fine nylon mesh, all the protoplasts were ruptured, leaving the fragile amyloplasts largely intact. Distribution of enzymes of glycolysis in plastids and soluble phase of sycamore protoplasts indicated that the absolute maximum activity for each glycolytic enzyme under optimum conditions exceeded the estimates of the maximal rate at which sycamore cells oxidize triose phosphate. Passage of protoplasts through the fine nylon mesh produced a 3-5-fold decrease in O2 consumption. However, addition of saturating amounts of respiratory substrates and ADP restored an O2 consumption equal to that observed with uncoupled intact protoplasts. Taken together, these results demonstrated that neither the overall capacity of the glycolytic enzymes in sycamore cells nor the availability of respiratory substrates for the mitochondria is ultimately responsible for determining the rate of uncoupled respiration in sycamore cells.

  16. Space Magnets Attracting Interest on Earth: Applications of Physical and Biological Techniques In the Study of Gravisensing and Response System of Plants (United States)

    Hasenstein, Karl H.; Boody, April; Cox, David (Technical Monitor)


    The BioTube/Magnetic Field Apparatus (MFA) research is designed to provide insight into the organization and operation of the gravity sensing systems of plants and other small organisms. This experiment on STS-107 uses magnetic fields to manipulate sensory cells in plant roots, thus using magnetic fields as a tool to study gravity-related phenomena. The experiment will be located in the SPACEHAB module and is about the size of a household microwave oven. The goal of the experiment is to improve our understanding of the basic phenomenon of how plants respond to gravity. The BioTube/MFA experiment specifically examines how gravitational forces serve as a directional signal for growth in the low-gravity environment of space. As with all basic research, this study will contribute to an improved understanding of how plants grow and will have important implications for improving plant growth and productivity on Earth. In BioTube/MFA, magnetic fields will be used to determine whether the distribution of subcellular starch grains, called amyloplasts, within plant cells predicts the direction in which roots will grow and curve in microgravity.

  17. Acid Phosphatase Activity May Affect the Tuber Swelling by Partially Regulating Sucrose-mediated Sugar Resorption in Potato

    Institute of Scientific and Technical Information of China (English)

    Da-Yong Wang; Yong Lian; De-Wei Zhu


    APase activity is involved in regulating many physiological and developmental events by affecting the resorption process.In this study, we investigate the role of APase activity in tuber development in potato. APase activities were mainly localized in cytoplasm, gaps among cells and stroma of amyloplasts of parenchyma cells at the stage of tuber swelling. AP1, encoding a putative APase, was also highly expressed in swelling tubers and a low level of expression was observed in elongated stolons and matured tubers. Inhibition of APase activity by applying Brefeldin A, an inhibitor of APase production and secretion, significantly suppressed the tuber swelling and moderately affected the stolon elongation and the tuberization frequency. During tuber development, sucrose serves as the main soluble sugar for long-distance transportation and resorption. Moreover, Inhibition of APase activity by Brefeldin A markedly reduced the sucrose content in tubers and further decreased the starch accumulation, suggesting that the function of APase in regulating the tuber swelling might be at least artially mediated by the sugar resorption. Exogenous sucrose treatments further indicate the important role of sucrose-mediated sugar resorption in tuber swelling. These results suggest that the APase activity might affect the tuber swelling by partially regulating the sucrose-mediated sugar resorption.

  18. Genetic controls on starch amylose content in wheat and rice grains

    Indian Academy of Sciences (India)

    Parviz Fasahat; Sadequr Rahman; Wickneswari Ratnam


    Starch accumulates in plants as granules in chloroplasts of source organs such as leaves (transitory starch) or in amyloplasts of sink organs such as seeds, tubers and roots (storage starch). Starch is composed of two types of glucose polymers: the essentially linear polymer amylose and highly branched amylopectin. The amylose content of wheat and rice seeds is an important quality trait, affecting the nutritional and sensory quality of two of the world’s most important crops. In this review, we focus on the relationship between amylose biosynthesis and the structure, physical behaviour and functionality of wheat and rice grains. We briefly describe the structure and composition of starch and then in more detail describe what is known about the mechanism of amylose synthesis and how the amount of amylose in starch might be controlled. This more specifically includes analysis of GBSS alleles, the relationship between waxy allelic forms and amylose, and related quantitative trait loci. Finally, different methods for increasing or lowering amylose content are evaluated.

  19. Arabidopsis ANGULATA10 is required for thylakoid biogenesis and mesophyll development. (United States)

    Casanova-Sáez, Rubén; Mateo-Bonmatí, Eduardo; Kangasjärvi, Saijaliisa; Candela, Héctor; Micol, José Luis


    The chloroplasts of land plants contain internal membrane systems, the thylakoids, which are arranged in stacks called grana. Because grana have not been found in Cyanobacteria, the evolutionary origin of genes controlling the structural and functional diversification of thylakoidal membranes in land plants remains unclear. The angulata10-1 (anu10-1) mutant, which exhibits pale-green rosettes, reduced growth, and deficient leaf lateral expansion, resulting in the presence of prominent marginal teeth, was isolated. Palisade cells in anu10-1 are larger and less packed than in the wild type, giving rise to large intercellular spaces. The ANU10 gene encodes a protein of unknown function that localizes to both chloroplasts and amyloplasts. In chloroplasts, ANU10 associates with thylakoidal membranes. Mutant anu10-1 chloroplasts accumulate H2O2, and have reduced levels of chlorophyll and carotenoids. Moreover, these chloroplasts are small and abnormally shaped, thylakoidal membranes are less abundant, and their grana are absent due to impaired thylakoid stacking in the anu10-1 mutant. Because the trimeric light-harvesting complex II (LHCII) has been reported to be required for thylakoid stacking, its levels were determined in anu10-1 thylakoids and they were found to be reduced. Together, the data point to a requirement for ANU10 for chloroplast and mesophyll development.

  20. Mechanism of dynamic reorientation of cortical microtubules due to mechanical stress. (United States)

    Muratov, Alexander; Baulin, Vladimir A


    Directional growth caused by gravitropism and corresponding bending of plant cells has been explored since 19th century, however, many aspects of mechanisms underlying the perception of gravity at the molecular level are still not well known. Perception of gravity in root and shoot gravitropisms is usually attributed to gravisensitive cells, called statocytes, which exploit sedimentation of macroscopic and heavy organelles, amyloplasts, to sense the direction of gravity. Gravity stimulus is then transduced into distal elongation zone, which is several mm far from statocytes, where it causes stretching. It is suggested that gravity stimulus is conveyed by gradients in auxin flux. We propose a theoretical model that may explain how concentration gradients and/or stretching may indirectly affect the global orientation of cortical microtubules, attached to the cell membrane and induce their dynamic reorientation perpendicular to the gradients. In turn, oriented microtubule arrays direct the growth and orientation of cellulose microfibrils, forming part of the cell external skeleton and determine the shape of the cell. Reorientation of microtubules is also observed in reaction to light in phototropism and mechanical bending, thus suggesting universality of the proposed mechanism. PMID:26422460

  1. Complex physiological and molecular processes underlying root gravitropism (United States)

    Chen, Rujin; Guan, Changhui; Boonsirichai, Kanokporn; Masson, Patrick H.


    Gravitropism allows plant organs to guide their growth in relation to the gravity vector. For most roots, this response to gravity allows downward growth into soil where water and nutrients are available for plant growth and development. The primary site for gravity sensing in roots includes the root cap and appears to involve the sedimentation of amyloplasts within the columella cells. This process triggers a signal transduction pathway that promotes both an acidification of the wall around the columella cells, an alkalinization of the columella cytoplasm, and the development of a lateral polarity across the root cap that allows for the establishment of a lateral auxin gradient. This gradient is then transmitted to the elongation zones where it triggers a differential cellular elongation on opposite flanks of the central elongation zone, responsible for part of the gravitropic curvature. Recent findings also suggest the involvement of a secondary site/mechanism of gravity sensing for gravitropism in roots, and the possibility that the early phases of graviresponse, which involve differential elongation on opposite flanks of the distal elongation zone, might be independent of this auxin gradient. This review discusses our current understanding of the molecular and physiological mechanisms underlying these various phases of the gravitropic response in roots.

  2. Systematic Analysis of Pericarp Starch Accumulation and Degradation during Wheat Caryopsis Development.

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    Xurun Yu

    Full Text Available Although wheat (Triticum aestivum L. pericarp starch granule (PSG has been well-studied, our knowledge of its features and mechanism of accumulation and degradation during pericarp growth is poor. In the present study, developing wheat caryopses were collected and starch granules were extracted from their pericarp to investigate the morphological and structural characteristics of PSGs using microscopy, X-ray diffraction and Fourier transform infrared spectroscopy techniques. Relative gene expression levels of ADP-glucose pyrophosphorylase (APGase, granule-bound starch synthase II (GBSS II, and α-amylase (AMY were quantified by quantitative real-time polymerase chain reaction. PSGs presented as single or multiple starch granules and were synthesized both in the amyloplast and chloroplast in the pericarp. PSG degradation occurred in the mesocarp, beginning at 6 days after anthesis. Amylose contents in PSGs were lower and relative degrees of crystallinity were higher at later stages of development than at earlier stages. Short-range ordered structures in the external regions of PSGs showed no differences in the developing pericarp. When hydrolyzed by α-amylase, PSGs at various developmental stages showed high degrees of enzymolysis. Expression levels of AGPase, GBSS II, and AMY were closely related to starch synthesis and degradation. These results help elucidate the mechanisms of accumulation and degradation as well as the functions of PSG during wheat caryopsis development.

  3. Β-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds. (United States)

    Srivastava, Garima; Kayastha, Arvind M


    Fenugreek (Trigonella foenum-graecum) seeds do not contain starch as carbohydrate reserve. Synthesis of starch is initiated after germination. A β-amylase from ungerminated fenugreek seeds was purified to apparent electrophoretic homogeneity. The enzyme was purified 210 fold with specific activity of 732.59 units/mg. Mr of the denatured enzyme as determined from SDS-PAGE was 58 kD while that of native enzyme calculated from size exclusion chromatography was 56 kD. Furthermore, its identity was confirmed to be β-amylase from MALDI-TOF analysis. The optimum pH and temperature was found to be 5.0 and 50°C, respectively. Starch was hydrolyzed at highest rate and enzyme showed a Km of 1.58 mg/mL with it. Antibodies against purified Fenugreek β-amylase were generated in rabbits. These antibodies were used for localization of enzyme in the cotyledon during different stages of germination using fluorescence and confocal microscopy. Fenugreek β-amylase was found to be the major starch degrading enzyme depending on the high amount of enzyme present as compared to α-amylase and also its localization at the periphery of amyloplasts. A new finding in terms of its association with protophloem was observed. Thus, this enzyme appears to be important for germination of seeds. PMID:24551136

  4. Reduced gravitropic sensitivity in roots of a starch-deficient mutant of Nicotiana sylvestris (United States)

    Kiss, J. Z.; Sack, F. D.


    Gravitropism was studied in seedlings of Nicotiana sylvestris Speg. et Comes wild-type (WT) and mutant NS 458 which has a defective plastid phosphoglucomutase (EC Starch was greatly reduced in NS 458 compared to the WT, but small amounts of starch were detected in rootcap columella cells in NS 458 by light and electron microscopy. The roots of WT are more sensitive to gravity than mutant NS 458 roots since: (1) in mutant roots, curvature was reduced and delayed in the time course of curvature; (2) curvature of mutant roots was 24-56% that of WT roots over the range of induction periods tested; (3) in intermittent-stimulation experiments, curvature of mutant roots was 37% or less than that of WT roots in all treatments tested. The perception time, determined by intermittent-stimulation experiments, was Planta 177, 198-206) support the conclusions that a full complement of starch is necessary for full gravitropic sensitivity and that amyloplasts function in gravity perception. Since a presumed relatively small increase in plastid buoyant mass (N. sylvestris mutant versus Arabidopsis mutant) significantly improves the orientation of the N. sylvestris mutant roots, we suggest that plastids are the likeliest candidates to be triggering gravity perception in roots of both mutants.

  5. Signal perception, transduction, and response in gravity resistance. Another graviresponse in plants (United States)

    Hoson, T.; Saito, Y.; Soga, K.; Wakabayashi, K.

    Resistance to the gravitational force is a serious problem that plants have had to solve to survive on land. Mechanical resistance to the pull of gravity is thus a principal graviresponse in plants, comparable to gravitropism. Nevertheless, only limited information has been obtained for this gravity response. We have examined the mechanism of gravity-induced mechanical resistance using hypergravity conditions produced by centrifugation. As a result, we have clarified the outline of the sequence of events leading to the development of mechanical resistance. The gravity signal may be perceived by mechanoreceptors (mechanosensitive ion channels) on the plasma membrane and it appears that amyloplast sedimentation in statocytes is not involved. Transformation and transduction of the perceived signal may be mediated by the structural or physiological continuum of microtubule-cell membrane-cell wall. As the final step in the development of mechanical resistance, plants construct a tough body by increasing cell wall rigidity. The increase in cell wall rigidity is brought about by modification of the metabolism of certain wall constituents and modification of the cell wall environment, especially pH. We need to clarify the details of each step by future space and ground-based experiments.

  6. Phytotoxicity of cobalt ions on the duckweed Lemna minor - Morphology, ion uptake, and starch accumulation. (United States)

    Sree, K Sowjanya; Keresztes, Áron; Mueller-Roeber, Bernd; Brandt, Ronny; Eberius, Matthias; Fischer, Wolfgang; Appenroth, Klaus-J


    Cobalt (Co2+) inhibits vegetative growth of Lemna minor gradually from 1 μM to 100 μM. Fronds accumulated up to 21 mg Co2+ g(-1) dry weight at 10 μM external Co2+ indicating hyperaccumulation. Interestingly, accumulation of Co2+ did not decrease the iron (Fe) content in fronds, highlighting L. minor as a suitable system for studying effects of Co2+ undisturbed by Fe deficiency symptoms unlike most other plants. Digital image analysis revealed the size distribution of fronds after Co2+ treatment and also a reduction in pigmentation of newly formed daughter fronds unlike the mother fronds during the 7-day treatment. Neither chlorophyll nor photosystem II fluorescence changed significantly during the initial 4d, indicating effective photosynthesis. During the later phase of the 7-day treatment, however, chlorophyll content and photosynthetic efficiency decreased in the Co2+-treated daughter fronds, indicating that Co2+ inhibits the biosynthesis of chlorophyll rather than leading to the destruction of pre-existing pigment molecules. In addition, during the first 4d of Co2+ treatment starch accumulated in the fronds and led to the transition of chloroplasts to chloro-amyloplasts and amylo-chloroplasts, while starch levels strongly decreased thereafter. PMID:25840119

  7. Nectar defense and hydrogen peroxide in floral nectar of Cucurbita pepo

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    Daniele Nocentini


    Full Text Available This study was carried out to investigate some similarities between the nectaries of Nicotiana sp. and Cucurbita pepo, such as starch accumulation in the nectary parenchyma, changes in nectary color during maturation, and the production of a large quantity of sucrose-dominant nectar. The concentration of hydrogen peroxide in C. pepo floral nectar was determined in order to verify the presence of a defense mechanism similar to that found in Nicotiana sp. which protects nectar from yeast and bacteria proliferation. We also tested the eventual accumulation of antioxidants in the nectary of C. pepo as a protection against oxidative stress caused by hydrogen peroxide. The level of hydrogen peroxide found in the floral nectar of C. pepo was much lower than that found in Nicotiana sp. and the male flowers of Cucurbita had a higher concentration than the female flowers. The low oxidative stress induced by this level of hydrogen peroxide caused the accumulation of a low amount of lutein inside the plastoglobules which were contained in amyloplasts. Plastids of the C. pepo nectary are specialized in the accumulation of starch rather than antioxidants.

  8. Auxin, ethylene and light in gravitropic growth: new insights (United States)

    Edelmann, Hg; Sabovljevic, A.; Njio, G.; Roth, U.

    The regulation mechanism of gravitropic differential plant growth is commonly divided into three sequential processes: the perception of the gravistimulus (generally attributed to amyloplast sedimentation), the transduction of the perceived signal (of which very little is known), and the adequate differential growth response (generally attributed to asymmetric auxin redistribution). The detailled mechanism is still unresolved and remains to be elucidated in significant parts. Employing 2D SDS-PAGE /Q-TOF amongst other methods and strategies we studied the effect of different auxins on gravitropism of coleoptiles and hypocotyls. We also analyzed the effects of light and ethylene (synthesis and perception) on gravitropic growth of primary shoots and roots and analyzed the protein pattern with respect to the observed physiological effects. In coleoptiles, under the applied experimental conditions the effect of 2,4-dichlorophenoxy acetic acid (2,4 D) on gravitropism differed from the effect of indolylacetic acid (IAA), which was similar to the one observed in sunflower hypocotyls. In roots, the relevance of ethylene for gravitropic differential growth and the capacity to evade mechanical barriers during horizontal gravistimulation was analyzed in detail. A special focus was addressed on the physiological significance of the root cap. We will show that the relevance of ethylene for gravitropism has hitherto been misjudged. Further new findings and their implications for the regulation mechanism of gravitropism will be presented and discussed. Kramer et al., (2003) J. Ex. Bot. 54, (393), 2723-2732 Edelmann, H.G., (2002) J. Ex. Bot. 53, (375), 1825-1828

  9. Characterization of chromoplasts and carotenoids of red- and yellow-fleshed papaya (Carica papaya L.). (United States)

    Schweiggert, Ralf M; Steingass, Christof B; Heller, Annerose; Esquivel, Patricia; Carle, Reinhold


    Chromoplast morphology and ultrastructure of red- and yellow-fleshed papaya (Carica papaya L.) were investigated by light and transmission electron microscopy. Carotenoid analyses by LC-MS revealed striking similarity of nutritionally relevant carotenoid profiles in both the red and yellow varieties. However, while yellow fruits contained only trace amounts of lycopene, the latter was found to be predominant in red papaya (51% of total carotenoids). Comparison of the pigment-loaded chromoplast ultrastructures disclosed tubular plastids to be abundant in yellow papaya, whereas larger crystalloid substructures characterized most frequent red papaya chromoplasts. Exclusively existent in red papaya, such crystalloid structures were associated with lycopene accumulation. Non-globular carotenoid deposition was derived from simple solubility calculations based on carotenoid and lipid contents of the differently colored fruit pulps. Since the physical state of carotenoid deposition may be decisive regarding their bioavailability, chromoplasts from lycopene-rich tomato fruit (Lycopersicon esculentum L.) were also assessed and compared to red papaya. Besides interesting analogies, various distinctions were ascertained resulting in the prediction of enhanced lycopene bioavailability from red papaya. In addition, the developmental pathway of red papaya chromoplasts was investigated during fruit ripening and carotenogenesis. In the early maturation stage of white-fleshed papaya, undifferentiated proplastids and globular plastids were predominant, corresponding to incipient carotenoid biosynthesis. Since intermediate plastids, e.g., amyloplasts or chloroplasts, were absent, chromoplasts are likely to emerge directly from proplastids.

  10. Isolation of New Gravitropic Mutants under Hypergravity Conditions (United States)

    Mori, Akiko; Toyota, Masatsugu; Shimada, Masayoshi; Mekata, Mika; Kurata, Tetsuya; Tasaka, Masao; Morita, Miyo T.


    Forward genetics is a powerful approach used to link genotypes and phenotypes, and mutant screening/analysis has provided deep insights into many aspects of plant physiology. Gravitropism is a tropistic response in plants, in which hypocotyls and stems sense the direction of gravity and grow upward. Previous studies of gravitropic mutants have suggested that shoot endodermal cells in Arabidopsis stems and hypocotyls are capable of sensing gravity (i.e., statocytes). In the present study, we report a new screening system using hypergravity conditions to isolate enhancers of gravitropism mutants, and we also describe a rapid and efficient genome mapping method, using next-generation sequencing (NGS) and single nucleotide polymorphism (SNP)-based markers. Using the endodermal-amyloplast less 1 (eal1) mutant, which exhibits defective development of endodermal cells and gravitropism, we found that hypergravity (10 g) restored the reduced gravity responsiveness in eal1 hypocotyls and could, therefore, be used to obtain mutants with further reduction in gravitropism in the eal1 background. Using the new screening system, we successfully isolated six ene (enhancer of eal1) mutants that exhibited little or no gravitropism under hypergravity conditions, and using NGS and map-based cloning with SNP markers, we narrowed down the potential causative genes, which revealed a new genetic network for shoot gravitropism in Arabidopsis.

  11. Post-harvest light treatment increases expression levels of recombinant proteins in transformed plastids of potato tubers. (United States)

    Larraya, Luis M; Fernández-San Millán, Alicia; Ancín, María; Farran, Inmaculada; Veramendi, Jon


    Plastid genetic engineering represents an attractive system for the production of foreign proteins in plants. Although high expression levels can be achieved in leaf chloroplasts, the results for non-photosynthetic plastids are generally discouraging. Here, we report the expression of two thioredoxin genes (trx f and trx m) from the potato plastid genome to study transgene expression in amyloplasts. As expected, the highest transgene expression was detected in the leaf (up to 4.2% of TSP). The Trx protein content in the tuber was approximately two to three orders of magnitude lower than in the leaf. However, we demonstrate that a simple post-harvest light treatment of microtubers developed in vitro or soil-grown tubers induces up to 55 times higher accumulation of the recombinant protein in just seven to ten days. After the applied treatment, the Trx f levels in microtubers and soil-grown tubers increased to 0.14% and 0.11% of TSP, respectively. Moreover, tubers stored for eight months maintained the capacity of increasing the foreign protein levels after the light treatment. Post-harvest cold induction (up to five times) at 4°C was also detected in microtubers. We conclude that plastid transformation and post-harvest light treatment could be an interesting approach for the production of foreign proteins in potato. PMID:26121393

  12. Proteomic alterations in root tips of Arabidopsis thaliana seedlings under altered gravity conditions (United States)

    Zheng, H. Q.; Wang, H.

    Gravity has a profound influence on plant growth and development Removed the influence of gravitational acceleration by spaceflight caused a wide range of cellular changes in plant Whole seedling that germinated and grown on clinostats showed the absent of gravitropism At the cellular level clinostat treatment has specific effects on plant cells such as induce alterations in cell wall composition increase production of heat-soluble proteins impact on the cellular energy metabolism facilitate a uniform distribution of plastids amyloplasts and increase number and volume of nucleoli A number of recent studies have shown that the exposure of Arabidopsis seedlings and callus cells to gravity stimulation hyper g-forces or clinostat rotation induces alterations in gene expression In our previous study the proteome of the Arabidopsis thaliana callus cells were separated by high resolution two-dimensional electrophoresis 2-DE Image analysis revealed that 80 protein spots showed quantitative and qualitative variations after exposure to clinostat rotation treatment We report here a systematic proteomic approach to investigate the altered gravity responsive proteins in root tip of Arabidopsis thaliana cv Landsberg erecta Three-day-old seedlings were exposed for 12h to a horizontal clinostat rotation H simulated weightlessness altered g-forces by centrifugation 7g hypergravity a vertical clinostat rotation V clinostat control or a stationary control grown conditions Total proteins of roots were extracted

  13. Role of phosphorylase in the mechanism of potato minituber storage cell changes during clinorotation (United States)

    Nedukha, O.; Shnyukova, E.

    The differences between the cytochemical reaction intensity and activity of phosphorylase (EC and carbohydrate content in storage parenchyma cells of Solanum tuberosum L. (cv Adreta) minitubers grown for 30 days in the horizontal clinostate (2 rev/min) and in the control have been studied by electroncytochemical and biochemical methods. It is established an acceleration of minitubers formation and storage parenchyma cell differentiation at clinorotation. Electroncytochemical investigation of phosphorylase activity localization in the storage parenchyma cells of minitubers grown in control and at clinorotation showed the product of the reaction as electron-dense precipitate was marked plastids. Intensity and density of precipitate was increase in stroma of plastids and on starch grain surface during of intensive growth of starch in amyloplast (on 10- and 20-days of the minituber formation) of clinorotated minitubers in comparison with that in the control. The precipitate amount was decreased in the plastids on 30 day of growth in both variants. Using biochemical methods it is found that activity of phosphorylase and content of mono- and disaccharide and also starch content changed in minitubers formed during clinorotation and in the control. Data obtained are discussed regarding the possible mechanism of phosphorylase activity change and the role of mono- and disaccharide in acceleration of storage organ formation during clinorotation.

  14. Plastid sedimentation kinetics in roots of wild-type and starch-deficient mutants of Arabidopsis (United States)

    MacCleery, S. A.; Kiss, J. Z.


    Sedimentation and movement of plastids in columella cells of the root cap were measured in seedlings of wild-type, a reduced starch mutant, and a starchless mutant of Arabidopsis. To assay for sedimentation, we used both linear measurements and the change of angle from the cell center as indices in vertical and reoriented plants with the aid of computer-assisted image analysis. Seedlings were fixed at short periods after reorientation, and plastid sedimentation correlated with starch content in the three strains of Arabidopsis. Amyloplasts of wild-type seedlings showed the greatest sedimentation, whereas plastids of the starchless mutant showed no significant sedimentation in the vertically grown and reoriented seedlings. Because previous research has shown that a full complement of starch is needed for full gravitropic sensitivity, this study correlates increased sensitivity with plastid sedimentation. However, although plastid sedimentation contributed to gravisensitivity, it was not required, because the gravitropic starchless mutant had plastids that did not sediment. This is the first study, to our knowledge, to measure plastid sedimentation in Arabidopsis roots after reorientation of seedlings. Taken together, the results of this study are consistent with the classic plastid-based and protoplast-based models of graviperception and suggest that multiple systems of perception exist in plant cells.

  15. Female gametophyte and pollen tube of Epilobium palustre L.

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    Józef Bednara


    Full Text Available The monosporic, tetranucleate embryo sac of Epilobium palustre (Onagraceae develops from the micropylar megaspore in a linear tetrad. In mononucleate embryo sacs a peculiar chromatic structure associated with a nucleolus appears in the nucleus. This structure seems to be formed by fibrillar material and is not visible in the subsequant stages of development. A large amount of rough ER cisternae occurs in the late mononucleate stage, during the binucleate stage their contents become optically dense. It the early tetranucleate stage the amount of ER is small, it increases again in the developing synergids and central cell. Numerous amyloplasts present in the mononucleate embryo sac loose their starch grains and some are transformed into cup-shaped plastids or proplastids. They are passed on to each of the embryo sac cells. The growth of the pollen tube ceases immediately after the penetration through the filiform apparatus of a synergid. At the apex of the tube a pore is formed. At the last stages of the penetration the apical part of the pollen tube becomes separated by a transverse partition from the distal part of the tube. The contents of the both parts differ in their internal structure. The distal part contains cytoplasm with numerous organoids, while the apical part is mainly filled with spherical bodies.

  16. Microtubules restrict plastid sedimentation in protonemata of the moss Ceratodon (United States)

    Schwuchow, J.; Sack, F. D.


    Apical cells of protonemata of the moss Ceratodon purpureus are unusual among plant cells with sedimentation in that only some amyloplasts sediment and these do not fall completely to the bottom of vertical cells. To determine whether the cytoskeleton restricts plastid sedimentation, the effects of amiprophos-methyl (APM) and cytochalasin D (CD) on plastid position were quantified. APM treatments of 30-60 min increased the plastid sedimentation that is normally seen along the length of untreated or control cells. Longer APM treatments often resulted in more dramatic plastid sedimentation, and in some cases almost all plastids sedimented to the lowermost point in the cell. In contrast, the microfilament inhibitor CD did not affect longitudinal plastid sedimentation compared to untreated cells, although it did disturb or eliminate plastid zonation in the tip. These data suggest that microtubules restrict the sedimentation of plastids along the length of the cell and that microtubules are load-bearing for all the plastids in the apical cell. This demonstrates the importance of the cytoskeleton in maintaining organelle position and cell organization against the force of gravity.

  17. Systematic Analysis of Pericarp Starch Accumulation and Degradation during Wheat Caryopsis Development. (United States)

    Yu, Xurun; Li, Bo; Wang, Leilei; Chen, Xinyu; Wang, Wenjun; Wang, Zhong; Xiong, Fei


    Although wheat (Triticum aestivum L.) pericarp starch granule (PSG) has been well-studied, our knowledge of its features and mechanism of accumulation and degradation during pericarp growth is poor. In the present study, developing wheat caryopses were collected and starch granules were extracted from their pericarp to investigate the morphological and structural characteristics of PSGs using microscopy, X-ray diffraction and Fourier transform infrared spectroscopy techniques. Relative gene expression levels of ADP-glucose pyrophosphorylase (APGase), granule-bound starch synthase II (GBSS II), and α-amylase (AMY) were quantified by quantitative real-time polymerase chain reaction. PSGs presented as single or multiple starch granules and were synthesized both in the amyloplast and chloroplast in the pericarp. PSG degradation occurred in the mesocarp, beginning at 6 days after anthesis. Amylose contents in PSGs were lower and relative degrees of crystallinity were higher at later stages of development than at earlier stages. Short-range ordered structures in the external regions of PSGs showed no differences in the developing pericarp. When hydrolyzed by α-amylase, PSGs at various developmental stages showed high degrees of enzymolysis. Expression levels of AGPase, GBSS II, and AMY were closely related to starch synthesis and degradation. These results help elucidate the mechanisms of accumulation and degradation as well as the functions of PSG during wheat caryopsis development. PMID:26394305

  18. Early root cap development and graviresponse in white clover (Trifolium repens) grown in space and on a two-axis clinostat. (United States)

    Smith, J D; Staehelin, L A; Todd, P


    White clover (Trifolium repens) was germinated and grown in microgravity aboard the Space Shuttle (STS-60, 1994; STS-63, 1995), on Earth in stationary racks and in a slow-rotating two-axis clinostat. The objective of this study was to determine if normal root cap development and early plant gravity responses were dependent on gravitational cues. Seedlings were germinated in space and chemically fixed in orbit after 21, 40, and 72 h. Seedlings 96 h old were returned viable to earth. Germination and total seedling length were not dependent on gravity treatment. In space-flown seedlings, the number of cell stories in the root cap and the geometry of central columella cells did not differ from those of the Earth-grown seedlings. The root cap structure of clinorotated plants appeared similar to that of seedlings from microgravity, with the exception of three-day rotated plants, which displayed significant cellular damage in the columella region. Nuclear polarity did not depend on gravity; however, the positions of amyloplasts in the central columella cells were dependent on both the gravity treatment and the age of the seedlings. Seedlings from space, returned viable to earth, responded to horizontal stimulation as did 1 g controls, but seedlings rotated on the clinostat for the same duration had a reduced curvature response. This study demonstrates that initial root cap development is insensitive to either chronic clinorotation or microgravity. Soon after differentiation, however, clinorotation leads to loss of normal root cap structure and plant graviresponse while microgravity does not. PMID:11543182

  19. Observation of fiber ultrastructure of Ligon lintless mutant in upland cotton during fiber elongation

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    CHENG Chaohua; WANG Xuede; NI Xiyuan


    Lintless mutant is a super-short fiber mutant in upland cotton only 4-8 mm in fiber length and also named Ligon cotton controlled by one dominant gene Li1. Fiber ultrastructure of the mutant (Li1) and its wild type (li1) in situ and in vitro was observed under an electron microscope to understand its cytological characteristics during the fiber cell elongation. The results showed that the mutant fiber in situ had thinner cytoplasm, more small vacuoles, less mitochondria, Golgi apparatus and endoplasmic reticula, and there were more starch granules which were free or packed in the amyloplast beside the cell wall than that of wild type. It was indicated that scarcity of functional organelles and disability of transformation from starch to sugar might be associated with the fact that the mutant fiber cell was aborted too early to elongate into normal length. Mutant ovule in some media containing GA3 could produce a kind of huge callus that grew faster than normal ovules. The callus was covered with many white, loose, and semitransparent fiber-like cells that apt to get off from ovule. These fiber-like cells were multicellular fibers generated by cell division and had black dots just like pigment glands in the stem and leaf of cotton. There were lots of micro-tubes beside cytoplasm membrane of the multicellular fiber, which were thought to be primary preparation for second wall deposition of multicellular fiber. It was indicated that GA3 might induce the expression of gene(s) that kept inactive in the field condition and then stimulate the original fiber cell in vitro to undergo division again.

  20. A bird’s-eye view of molecular changes in plant gravitropism using omics techniques

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    Oliver eSchüler


    Full Text Available During evolution, plants have developed mechanisms to adapt to a variety of environmental stresses, including drought, high salinity, changes in carbon dioxide levels and pathogens. Central signaling hubs and pathways that are regulated in response to these stimuli have been identified. In contrast to these well studied environmental stimuli, changes in transcript, protein and metabolite levels in response to a gravi¬tational stimulus are less well understood. Amyloplasts, localized in statocytes of the root tip, in meso¬phyll cells of coleoptiles and in the elongation zone of the growing internodes comprise statoliths in higher plants. Deviations of the statocytes with respect to the earthly gravity vector lead to a displacement of statoliths relative to the cell due to their inertia and thus to gravity perception. Down¬stream signaling events, including the conversion from the biophysical signal of sedi¬men¬tation of distinct heavy mass to a biochemical signal, however, remain elusive. More recently, technical advances, including clinostats, drop towers, parabolic flights, satellites and the International Space Station, allowed researchers to study the effect of altered gravity conditions - real and simulated micro- as well as hypergravity on plants. This allows for a unique opportunity to study plant responses to a purely anthropogenic stress for which no evolutionary program exists. Furthermore, the requirement for plants as food and oxygen sources during prolonged manned space explorations led to an increased interest in the identification of genes involved in the adaptation of plants to microgravity.Transcriptomic, proteomic, phosphoproteomic, and metabolomic profiling strategies provide a sensitive high-throughput approach to identify biochemical alterations in response to changes with respect to the influence of the gravitational vector and thus the acting gravitational force on the transcript, protein and metabolite level. This

  1. New Insights into Fe Localization in Plant Tissues

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    Hannetz eRoschzttardtz


    Full Text Available Deciphering cellular iron (Fe homeostasis requires having access to both quantitative and qualitative information on the subcellular pools of Fe in tissues and their dynamics within the cells. We have taken advantage of the Perls/DAB Fe staining procedure to perform a systematic analysis of Fe distribution in roots, leaves and reproductive organs of the model plant Arabidopsis thaliana, using wild-type and mutant genotypes affected in iron transport and storage. Roots of soil-grown plants accumulate iron in the apoplast of the central cylinder, a pattern that is strongly intensified when the citrate effluxer FRD3 is not functional, thus stressing the importance of citrate in the apoplastic movement of Fe. In leaves, Fe level is low and only detected in and around vascular tissues. In contrast, Fe staining in leaves of iron-treated plants extends in the surrounding mesophyll cells where Fe deposits, likely corresponding to Fe-ferritin complexes, accumulate in the chloroplasts. The loss of ferritins in the fer1,3,4 triple mutant provoked a massive accumulation of Fe in the apoplastic space, suggesting that in the absence of iron buffering in the chloroplast, cells activate iron efflux and/or repress iron influx to limit the amount of iron in the cell. In flowers, Perls/DAB staining has revealed a major sink for Fe in the anthers. In particular, developing pollen grains accumulate detectable amounts of Fe in small-size intracellular bodies that aggregate around the vegetative nucleus at the binuclear stage and that were identified as amyloplasts. In conclusion, using the Perls/DAB procedure combined to selected mutant genotypes, this study has established a reliable atlas of Fe distribution in the main Arabidopsis organs, proving and refining long-assumed intracellular locations and uncovering new ones. This iron map of Arabidopsis will serve as a basis for future studies of possible actors of iron movement in plant tissues and cell compartments.

  2. Gravity resistance, another graviresponse in plants - role of microtubule-membrane-cell wall continuum (United States)

    Hoson, T.; Saito, Y.; Usui, S.; Soga, K.; Wakabayashi, K.

    Resistance to the gravitational force has been a serious problem for plants to survive on land, after they first went ashore more than 400 million years ago. Thus, gravity resistance is the principal graviresponse in plants comparable to gravitropism. Nevertheless, only limited information has been obtained for this second gravity response. We have examined the mechanism of gravity resistance using hypergravity conditions produced by centrifugation. The results led a hypothesis on the mechanism of plant resistance to the gravitational force that the plant constructs a tough body by increasing the cell wall rigidity, which are brought about by modification of the cell wall metabolism and cell wall environment, especially pH. The hypothesis was further supported by space experiments during the Space Shuttle STS-95 mission. On the other hand, we have shown that gravity signal may be perceived by mechanoreceptors (mechanosensitive ion channels) on the plasma membrane and amyloplast sedimentation in statocytes is not involved in gravity resistance. Moreover, hypergravity treatment increased the expression levels of genes encoding alpha-tubulin, a component of microtubules and 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGR), which catalyzes a reaction producing mevalonic acid, a key precursor of terpenoids such as membrane sterols. The expression of HMGR and alpha- and beta-tubulin genes increased within several hours after hypergravity treatment, depending on the magnitude of gravity. The determination of levels of gene products as well as the analysis with knockout mutants of these genes by T-DNA insertions in Arabidopsis supports the involvement of both membrane sterols and microtubules in gravity resistance. These results suggest that structural or physiological continuum of microtubule-cell membrane-cell wall is responsible for plant resistance to the gravitational force.

  3. Pseudopollen in Eria Lindl. Section Mycaranthes Rchb.f. (Orchidaceae) (United States)

    DAVIES, K. L.; TURNER, M. P.


    • Background and Aims Pseudopollen is a whitish, mealy material produced upon the labella of a number of orchid species as labellar hairs either become detached or fragment. Since individual hair cells are rich in protein and starch, it has long been speculated that pseudopollen functions as a reward for visiting insects. Although some 90 years have passed since Beck first described pseudopollen for a small number of Eria spp. currently assigned to section Mycaranthes Rchb.f., we still know little about the character of pseudopollen in this taxon. The use of SEM and histochemistry would re-address this deficit in our knowledge whereas comparison of pseudopollen in Eria (S.E. Asia), Maxillaria (tropical and sub-tropical America), Polystachya (largely tropical Africa and Madagascar) and Dendrobium unicum (Thailand and Laos) would perhaps help us to understand better how this feature may have arisen and evolved on a number of different continents. • Methods Pseudopollen morphology is described using light microscopy and scanning electron microscopy. Hairs were tested for starch, lipid and protein using IKI, Sudan III and the xanthoproteic test, respectively. • Key Results and Conclusions The labellar hairs of all eight representatives of section Mycaranthes examined are identical. They are unicellular, clavate with a narrow ‘stalk’ and contain both protein and starch but no detectable lipid droplets. The protein is distributed throughout the cytoplasm and the starch is confined to amyloplasts. The hairs become detached from the labellar surface and bear raised cuticular ridges and flaky deposits that are presumed to be wax. In that they are unicellular and appear to bear wax distally, the labellar hairs are significantly different from those observed for other orchid species. Comparative morphology indicates that they evolved independently in response to pollinator pressures similar to those experienced by other unrelated pseudopollen-forming orchids on

  4. Identification and analysis of novel genes involved in gravitropism of Arabidopsis thaliana. (United States)

    Morita, Miyo T.; Tasaka, Masao; Masatoshi Taniguchi, .


    Gravitropism is a continuous control with regard to the orientation and juxtaposition of the various parts of the plant body in response to gravity. In higher plants, the relative directional change of gravity is mainly suscepted in specialized cells called statocytes, followed by signal conversion from physical information into physiological information within the statocytes. We have studied the early process of shoot gravitropism, gravity sensing and signaling process, mainly by molecular genetic approach. In Arabidopsis shoot, statocytes are the endodermal cells. sgr1/scarcrow (scr) and sgr7/short-root (shr) mutants fail to form the endodermis and to respond to gravity in their inflorescence stems. Since both SGR1/SCR and SGR7/SHR are transcriptional factors, at least a subset of their downstream genes can be expected to be involved in gravitropism. In addition, eal1 (endodermal-amyloplast less 1), which exhibits no gravitropism in inflorescence stem but retains ability to form endodermis, is a hypomorphic allele of sgr7/shr. Take advantage of these mutants, we performed DNA microarray analysis and compared gene expression profiles between wild type and the mutants. We found that approx. 40 genes were commonly down-regulated in these mutants and termed them DGE (DOWN-REGULATED GENE IN EAL1) genes. DGE1 has sequence similarity to Oryza sativa LAZY1 that is involved in shoot gravitropism of rice. DGE2 has a short region homologous to DGE1. DTL (DGE TWO-LIKE}) that has 54% identity to DGE2 is found in Arabidopsis genome. All three genes are conserved in angiosperm but have no known functional domains or motifs. We analyzed T-DNA insertion for these genes in single or multiple combinations. In dge1 dge2 dtl triple mutant, gravitropic response of shoot, hypocotyl and root dramatically reduced. Now we are carrying out further physiological and molecular genetic analysis of the triple mutant.

  5. Comparison Analysis of Starch and Protein Expression Profiles on Cassava Tuberous Rootscv. SC8 and Its Tetraploid%华南8号木薯及其四倍体块根淀粉及蛋白表达谱的差异分析

    Institute of Scientific and Technical Information of China (English)

    安飞飞; 陈松笔; 李庚虎; 周锴; 李开绵


    [Objective]The aim of the study was to better understand the change of starch content, starch structure and its proteins regulatory mechanisms of cassava tuberous roots of diploid and its tetraploid.[Method]Tuberous roots of 10 months after planting were used as the materials, starch content was measured by weight in air and water, the proportion of amylose and amylopectin was determined by spectrophotometry, Excel 2013 and DPS v7.05 statistical software were used to analyze the data, the significant difference was determined by using Duncan method, the size, shape and number of amyloplast were observed by scanning electron microscopy, Western blot was used to verify some enzymes involved in starch synthesis and degradation, roots proteins of cassava tetraploid and diploid genotypes were excised and purified from 2D gels, Delta 2D software were used to analyze the different proteins with more than 2.0 fold changes and assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-TOF-MS/MS) were used to identify them, and the proteins were classified according to their functions combined with KEGG database.[Result]After chromosome doubling in tetraploid cassava, dry matter content, starch content and fresh roots weight were significantly decreased by 12.18%, 11.41%, and 35.34%, respectively. No significant difference was observed in amylase and amylopectin proportion. No significant difference in amyloplast morphology, mainly spherical, irregular sphere and ellipsoid also exist, the size was uniform, while the loose degree of amyloplast and space gap were increased, the number was decreased. The level of sucrose phosphate synthase (SPS) significantly decreased,β-amylase expression was increased, while the level of granule bound starch synthase I (GBSSI) was not changed. Twenty differential expressed protein spots were showed and analyzed by software, including 2 up-regulated spots and 18 down-regulated spots, 19 were successfully matched in

  6. Root gravitropism: an experimental tool to investigate basic cellular and molecular processes underlying mechanosensing and signal transmission in plants (United States)

    Boonsirichai, K.; Guan, C.; Chen, R.; Masson, P. H.


    The ability of plant organs to use gravity as a guide for growth, named gravitropism, has been recognized for over two centuries. This growth response to the environment contributes significantly to the upward growth of shoots and the downward growth of roots commonly observed throughout the plant kingdom. Root gravitropism has received a great deal of attention because there is a physical separation between the primary site for gravity sensing, located in the root cap, and the site of differential growth response, located in the elongation zones (EZs). Hence, this system allows identification and characterization of different phases of gravitropism, including gravity perception, signal transduction, signal transmission, and curvature response. Recent studies support some aspects of an old model for gravity sensing, which postulates that root-cap columellar amyloplasts constitute the susceptors for gravity perception. Such studies have also allowed the identification of several molecules that appear to function as second messengers in gravity signal transduction and of potential signal transducers. Auxin has been implicated as a probable component of the signal that carries the gravitropic information between the gravity-sensing cap and the gravity-responding EZs. This has allowed the identification and characterization of important molecular processes underlying auxin transport and response in plants. New molecular models can be elaborated to explain how the gravity signal transduction pathway might regulate the polarity of auxin transport in roots. Further studies are required to test these models, as well as to study the molecular mechanisms underlying a poorly characterized phase of gravitropism that is independent of an auxin gradient.

  7. Structural development of aleurone and its function in common wheat. (United States)

    Xiong, Fei; Yu, Xu-Run; Zhou, Liang; Wang, Zhong; Wang, Feng; Xiong, Ai-Sheng


    The wheat aleurone is formed from surface endosperm cells, and its developmental status reflects its biogenesis, structural characteristics, and physiological functions. In this report, wheat caryopses at different development stages were embedded in Spurr's low-viscosity embedding medium for observation of the development of aleurone cells (ACs) by light microscopy, scanning electron microscopy, and fluorescence microscopy, respectively. According to their structures and physiological characterization, the ACs development process was divided into five stages: endosperm cellulization, spherosome formation, aleurone grain formation, filling material proliferation, and maturation. Furthermore, ACs in different parts of the caryopsis formed differently. ACs near the vascular bundle developed earlier and formed transfer cells, but other ACs formed slowly and did not form transfer cells. ACs on the caryopsis backside were a regular square shape; however, ACs in the caryopsis abdomen were mainly irregular. There were also differences in development between wheat varieties. ACs were rectangular in hard wheat but square in soft wheat. ACs were larger and showed a greater degree of filling in hard compared to soft wheat. The storage materials in ACs were different compared to inner endosperm cells (IECs). The concentrations of minerals such as sodium, magnesium, silicon, phosphorus and potassium were higher in ACs than in IECs. ACs contained many aleurone grains and spherosomes, which store lipids and mineral nutrients, respectively. The cell nucleus did not disappear and the cells were still alive during aleurone maturation. However, IECs were dead and mainly contained amyloplast and protein bodies, which store starch and protein, respectively. Overall, the above results characterized major structural features of aleurone and revealed that the wheat aleurone has mainly four functions.

  8. Structure and development of stomata on the primary root of Ceratonia siliqua L. (United States)

    Christodoulakis, N S; Menti, J; Galatis, B


    Stomata of various sizes are produced on the primary root of Ceratonia siliqua L. Most are generated during embryogenesis, prior to seed desiccation. They can be detected on the dry embryo in a wide zone just above the root tip. Initially, large stomata are formed. These have the ability to induce divisions of their neighbouring cells, creating particular cell patterns around them. Later, small perigenous stomata are generated. As the root grows following seed germination, the stomatal zone overlaps with that of the root hairs. Although root stomata of C. siliqua undergo a structural differentiation that seems almost identical to that of the elliptical stomata formed on leaves, they are unable to move and remain permanently open. Polarizing microscopy of fully differentiated stomata and young stomata at the stage of stomatal pore formation revealed deposition of radial cellulose microfibril systems on their periclinal walls. However, these systems were less developed than those on leaf stomata, a feature that might be responsible for their inactivity. Besides, plastids of the root guard cells (GCs) do not differentiate into chloroplasts but function solely as amyloplasts. Root stomata have a short life span. During rapid and intense root growth, GCs cannot keep pace with the elongation of their neighbouring rhizodermal cells. They therefore split in their mid-region, transversely to the stoma axis. The two parts of the transversely torn stoma are dragged apart and a large opening is formed on the root surface, just above the substomatal cavity. The root stomata, together with these openings, may facilitate increased gaseous exchange during respiration and/or an increased transfer of some nutrients and water in the rapidly growing primary root.

  9. Investigation of Endosperm Cell Development of Different Rice Varieties%不同类型水稻品种胚乳发育的研究

    Institute of Scientific and Technical Information of China (English)

    李栋梁; 李小刚; 顾蕴洁; 王忠


    不同部位的分布上,4种水稻品种各自呈现了一定程度的差异与联系。粳稻2个品种与籼稻2个品种间差异更多体现在颖果发育进程上;而糯稻与非糯稻的区别则更多体现在淀粉体的形态与灌浆的充实程度上。%The objective of this research is to clarify the difference of rice endosperm of different varieties during the development process.[Method]Four rice varieties, Nipponbare, Yangdao 6, Wuyunuo16 and Yangfunuo 4 were used as the experimental materials, and their caryopsis development days precisely were precisely recorded and their caryopsis development was closely observed. Starch accumulation and physiological activities were observed by I2-KI and TTC staining. Structure changes of rice starch endosperm cells were observed by applying spur resin embedding and semi-thin sectioning, and light microscopy. The ultrastructure and element analysis of full ripe rice caryopsis were observed under scanning electron microscope and EDS. [Result]Four stages occurred in caryopses development of all the rice varieties tested: formation stage, milky stage, dough stage, and full maturity stage. Compared to development stages of the endosperm, formation stage runs in parallel to the coenocyte and cellurization stages, which are the initial stage of endosperm formation; milky stage corresponds to the differentiation stage of endosperm cells, while the last two stages, dough and full maturity stages, are equivalent to that of the endosperm maturity stage. The shape of two rice sub subspecies, japonica and indica, varies significantly. Of the four rice varieties tested, two indica varieties showed a relatively faster growth rate than two japonica cultivars; Wuyunuo16 and Yangfunuo 4 had almost identical growth rate of dry matter accumulation. Nuclei of starch endosperm cells degenerated with the extension of amyloplasts, while amyloplasts were still enlarging and growing. Amyloplasts in the endosperm cells were compound granules, and their

  10. Himatanthus lancifolius (Müll. Arg. Woodson, Apocynaceae: estudo farmacobotânico de uma planta medicinal da Farmacopeia brasileira 1ª edição Himatanthus lancifolius (Müll. Arg. Woodson, Apocynaceae: morpho-anatomical study of a medicinal plant described in the Brazilian Pharmacopoeia 1st edition

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    Leopoldo C. Baratto


    Full Text Available Himatanthus lancifolius (Müll. Arg. Woodson é um arbusto nativo do Brasil, latescente, popularmente conhecido como agoniada e utilizado principalmente para distúrbios menstruais. O presente trabalho teve como objetivo caracterizar morfoanatomicamente a folha, o caule e a casca caulinar dessa planta medicinal, a fim de contribuir para o controle de qualidade e a autenticidade dessa espécie. O material vegetal foi fixado e submetido às microtécnicas usuais. A folha é simples, glabra e obovado-lanceolada. A epiderme é uniestratificada, revestida por cutícula estriada e possui estômatos anisocíticos na face abaxial. O mesofilo é dorsiventral. A nervura central é biconvexa e o pecíolo é circular, ambos apresentando feixes vasculares bicolaterais. Laticíferos, amiloplastos e idioblastos fenólicos estão presentes no parênquima fundamental da nervura central e do pecíolo. O sistema vascular do caule é tipicamente bicolateral. Laticíferos e idioblastos fenólicos ocorrem no córtex, no floema e na medula. Esses caracteres morfoanatômicos, em conjunto, podem ser utilizados como parâmetros para o controle de qualidade dessa espécie.Himatanthus lancifolius (Müll. Arg. Woodson is a Brazilian native shrub, laticiferous, popularly known as "agoniada" and it is mainly used for uterine disorders. The present work aimed to study the leaf, stem and stem bark morpho-anatomy of this medicinal plant, in order to contribute to its quality control and identification. The plant material was fixed and submitted to standard microtechniques. The leaf is simple, glabrous and obovate-lanceolate. The epidermis is uniseriate, coated with striated cuticle and it has anysocitic stomata on the abaxial surface. The mesophyll is dorsiventral. The midrib is biconvex and the petiole is circular, both presenting bicollateral vascular bundles. Laticiferous ducts, amyloplasts and phenolic idioblasts are found in ground parenchyma of the midrib and petiole. The

  11. Water mediated alterations in gravity signal transform phytofilertation capability in hydroponic plants (United States)

    Singh, Yogranjan; Singh Marabi, Rakesh; Satpute, Gyanesh Kumar; Mishra, Stuti


    signal is generated by the sedimentation of the amyloplasts. This induces a signal transduction pathway that promotes an auxin gradient across the root. The proteinogenic amino acid proline functions as a radical scavenger, electron sink, stabilizer of macromolecules, cell wall component and a metal chelation compound. In order to have most competent option for phytofilteration, the natural biodiversity out of aquatic ecosystem should be better studied. Screening of plants that produce natural chemicals whose structures are similar to the xenobiotic compounds should be the first step of any phytoremediation process. An experimental hydroponic-phytofilteration system with real effluent must give pragmatic information on the real detoxification capacity of the plants and allow determining the appropriate design and size of the future constructed wetland system to clean up the contaminated wastewater to reduce negative impact of eutrophication.

  12. ARG1 and ARL2 contribute to gravity signal transduction in the statocytes of Arabidopsis thaliana roots and hypocotyls (United States)

    Masson, Patrick; Harrison, Benjamin; Stanga, John; Otegui, Marisa; Sedbrook, John

    those of the single mutants. We used this observation to design a genetic screen for the identification of new loci that contribute to the pgm gravity-signaling pathway. Two genetic enhancers of arg1-2 were identified this way, called mar1-1 and mar2-1. These mutations were shown to affect components of the protein-import complex found in the outer membrane of plastids. Interestingly, the columellar amyloplasts of arg1-2 mar2-1 mutant roots display wild-type ultra-structure, accumulate starch and sediment at wild-type rates upon gravistimulation. We conclude that the plastid outer envelope may contribute directly to gravity signal transduction within the statocytes.

  13. Iron and ferritin accumulate in separate cellular locations in Phaseolus seeds

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    Blair Matthew W


    Full Text Available Abstract Background Iron is an important micronutrient for all living organisms. Almost 25% of the world population is affected by iron deficiency, a leading cause of anemia. In plants, iron deficiency leads to chlorosis and reduced yield. Both animals and plants may suffer from iron deficiency when their diet or environment lacks bioavailable iron. A sustainable way to reduce iron malnutrition in humans is to develop staple crops with increased content of bioavailable iron. Knowledge of where and how iron accumulates in seeds of crop plants will increase the understanding of plant iron metabolism and will assist in the production of staples with increased bioavailable iron. Results Here we reveal the distribution of iron in seeds of three Phaseolus species including thirteen genotypes of P. vulgaris, P. coccineus, and P. lunatus. We showed that high concentrations of iron accumulate in cells surrounding the provascular tissue of P. vulgaris and P. coccineus seeds. Using the Perls' Prussian blue method, we were able to detect iron in the cytoplasm of epidermal cells, cells near the epidermis, and cells surrounding the provascular tissue. In contrast, the protein ferritin that has been suggested as the major iron storage protein in legumes was only detected in the amyloplasts of the seed embryo. Using the non-destructive micro-PIXE (Particle Induced X-ray Emission technique we show that the tissue in the proximity of the provascular bundles holds up to 500 μg g-1 of iron, depending on the genotype. In contrast to P. vulgaris and P. coccineus, we did not observe iron accumulation in the cells surrounding the provascular tissues of P. lunatus cotyledons. A novel iron-rich genotype, NUA35, with a high concentration of iron both in the seed coat and cotyledons was bred from a cross between an Andean and a Mesoamerican genotype. Conclusions The presented results emphasize the importance of complementing research in model organisms with analysis in

  14. Ultrastructural features of Mimulus aurantiacus (Scrophulariaceae pollen tubes in vivo

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    Nuran Ekici


    Full Text Available The aim of this study is to give information on ultrastructure of in vivo pollen tubes of Mimulus aurantiacus which were collected from the Botanical Garden of the University of California at Berkeley. Materials were prepared according to electron microscopy methods and examined under Zeiss electron microscope. Four zones were examined in the pollen tubes of Mimulus aurantiacus. APICAL ZONE: Mitochondria, smooth endoplasmic reticulum, rough endoplasmic reticulum, dictyosomes and secretory vesicles were observed. SUBAPICAL ZONE: This area contained abundant rough endoplasmic reticulum and occasionally some smooth endoplasmic reticulum. The polysomes, mitochondria, proplastids that contain starch, small vacuoles and a few lipid bodies were detected. NUCLEAR ZONE: Both generative and vegetative cell nuclei lie in this zone. The vegetative cell nucleus was large and long. Rough endoplasmic reticulum, mitochondria, ribosomes, dictyosomes, and amyloplasts that are rich of starch were observed. VACUOLATION AND PLUG FORMATION ZONE: Cytoplasm of the tubes was full of large vacuoles. Few organelles such as mitochondria, dictyosome and rough endoplasmic reticulum were detected along their periphery.O objetivo deste estudo é informar sobre a ultraestrutura de tubos de pólen de Mimulus aurantiacus in vivo coletados no "Botanical Garden" da Universidade da Califórnia em Berkeley. O material foi preparado de acordo com os métodos de microscopia eletrônica e examinado em microscópio eletrônico Zeiss. Quatro zonas dos tubos de pólen de Mimulus aurantiacus foram examinadas. ZONA APICAL: foram observados mitocôndrias, retículo endoplasmático liso; retículo endoplasmático rugoso, dictiossomos e vesículas secretoras. ZONA SUBAPICAL: esta área continha retículo endoplasmático rugoso em abundância e, ocasionalmente, algum retículo endoplasmático liso. Foram detectados polissomos, mitocôndrias, proplastídeos que contêm amido, pequenos vacúolos e

  15. The potential of two Salix genotypes for radionuclide/heavy metal accumulation. A case study of Rovinari ash pit (Gorj District, Romania) (United States)

    Hernea, Cornelia; Neţoiu, Constantin; Corneanu, Gabriel; Crăciun, Constantin; Corneanu, Mihaela; Cojocaru, Luminiţa; Rovena Lăcătuşu, Anca; Popescu, Ion


    heavy metals and radionuclides, in comparison with Salix alba. In seedlings developed on ash waste dump, in leaf cells, the fine blocks of heterochromatin are dispersed in nucleus. The chloroplast with well developed grana and numerous plastoglobuls, are in active synthesis (being present 2 - 4 starch grains), some chloroplasts being transformed in amyloplast. In the mitochondria matrix, are present ferritin aggregates, with role in cell detoxification processes.

  16. Actin Cytoskeleton-Based Plant Synapse as Gravitransducer in the Transition Zone of the Root Apex (United States)

    Baluska, Frantisek; Barlow, Peter; Volkmann, Dieter; Mancuso, Stefano

    The actin cytoskeleton was originally proposed to act as the signal transducer in the plant gravity sensory-motoric circuit. Surprisingly, however, several studies have documented that roots perfom gravisensing and gravitropism more effectively if exposed to diverse anti-F-actin drugs. Our study, using decapped maize root apices, has revealed that depolymerization of F-actin stimulates gravity perception in cells of the transition zone where root gravitropism is initiated (Mancuso et al. 2006). It has been proposed (Balǔka et al. 2005, 2009a) that s the non-growing adhesive end-poles, enriched with F-actin and myosin VIII, and active in endocytic recycling of both PIN transporters and cell wall pectins cross-linked with calcium and boron, act as the gravisensing domains, and that these impinge directly upon the root motoric responses via control of polar auxin transport. This model suggests that mechanical asymmetry at these plant synapses determines vectorial gravity-controlled auxin transport. Due to the gravity-imposed mechanical load upon the protoplast, a tensional stress is also imposed upon the plasma membrane of the physically lower synaptic cell pole. This stress is then relieved by shifting the endocytosis-exocytosis balance towards exocytosis (Balǔka et al. s 2005, 2009a,b). This `Synaptic Auxin Secretion' hypothesis does not conflict with the `Starch Statolith' hypothesis, which is based on amyloplast sedimentation. In fact, the `Synaptic Auxin Secretion' hypothesis has many elements which allow its unification with the Starch-Statolith model (Balǔka et al. 2005, 2009a,b). s References Balǔka F, Volkmann D, Menzel D (2005) Plant synapses: actin-based adhesion s domains for cell-to-cell communication. Trends Plant Sci 10: 106-111 Balǔka F, Schlicht M, s Wan Y-L, Burbach C, Volkmann D (2009a) Intracellular domains and polarity in root apices: from synaptic domains to plant neurobiology. Nova Acta Leopoldina 96: 103-122 Balǔka s F, Mancuso S

  17. Caracterização morfoanatômica de raiz e rizoma de Symphytum officinale L. (Boraginaceae Morpho-anatomical characterization of the root and rhizome of Symphytum officinale L. (Boraginaceae

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    Ana Cristina O. Toledo


    Full Text Available Symphytum officinale L. (Boraginaceae é uma erva perene conhecida como confrei, consólida-maior e erva-do-cardeal, empregada na fitoterapia como antiinflamatório, emoliente e anestésico suave de uso tópico, devido à ação da alantoína encontrada nos órgãos subterrâneos e na folha. Com a finalidade de contribuir com a identificação da planta medicinal e da droga vegetal, foram realizados estudos morfoanatômicos de raiz e rizoma, em estrutura secundária. O material botânico foi fixado e preparado de acordo com microtécnicas usuais. Os órgãos subterrâneos são semelhantes, fusiformes e mostram-se de cor amarela clara a negra. A raiz e o rizoma apresentam periderme e tecidos vasculares secundários semelhantes, mas diferem pela presença de xilema primário exarco na raiz e xilema primário endarco e medula parenquimática no rizoma. Numerosas células parenquimáticas contendo mucilagem e amiloplastos são observadas na raiz e no rizoma.Symphytum officinale L. (Boraginaceae is a perennial herb known as comfrey, gum plant or boneset, and it is employed topically as anti-inflammatory, emollient and mild anesthetic in phytotherapy, due to allantoin found in the underground organs and leaf. In order to contribute to the medicinal plant and vegetal drug identification, morpho-anatomical investigations of the root and rhizome, in secondary growth, were carried out. The botanical material was fixed and prepared according to usual microtechniques. The underground organs are alike, slender and yellow to black coloured. The root and the rhizome show similar periderm and secondary vascular tissues, yet they differ in that the root shows exarch primary xylem, while the rhizome presents endarch primary xylem and parenchymatous pith. Numerous parenchymatic cells which contain mucilage and amyloplasts are observed in the root and rhizome.

  18. 酸铝胁迫下常绿杨根冠超微结构变化%Acid and Aluminum Stress on Root Cap Ultrastructure of Evergreen Poplar Clone

    Institute of Scientific and Technical Information of China (English)

    钱莲文; 吴文杰


    The climate of China's subtropical area is warm humid, abundant rainfall, is fast-grown forest base. Forest soil is partial acidic or acidic, rich in aluminum. Which easily cause aluminum poisoning for the plant.A-61/186 (Populus x euramericana CV.) is the semi-evergreen poplar clone cultivated by Australian doctor L.D.Pryor, which female parent is Populus deltoids Marsh from the southern United States, and the male parent is originated Populus nigra CV. of Chile. It is one of three evergreen poplar clones and suitable for tropical and subtropical areas were introduced from Pakistan in 1987 by Mr. Shikai Zheng, who is a researcher of state forestry administration P.R.China. The changs in Ultrastructure of root cap cells were studied under controlled concentration and acting time by evergreen poplar clone A-61/186 (Populus euramericana CV.), using hydroponic experiment acid aluminum stress. Root cap cell ultrastructure were observed and shooting under the transmission electron microscopein in 30 d, 60 d and 90 d. Acid concentration and acted time under aluminum stress on its ultrastructure were explored. Used for revealing changes of poplar root cap cells ultrastructure under aluminum toxicity, then for aluminum toxicity submicroscopic level evaluation of root cap cells. The results showed that cellwall was dissolved and brokend with the increasing of stress concentration, and Al content 3 was obvious . Cell matrix changed from gel state to Scattered debris. Starch grain declined in the number and amyloplasts increased in the number. While the core deviation of starch grain heap were changed from non-clear to clear trend. With the development of acting time, Starch grain were sharply reduced in Al content 2 under 60 d stresse and amyloplasts in Al conten 3 under 90 d stress. The core of starch grain heap was not deviated obviously.%中国的亚热带地区气候温暖湿润、雨量充沛,是中国速生丰产林基地,森林土壤多呈偏酸性和酸性,为

  19. ``Rhizogenesis in vitro'' - as a model to study microgravity biological effects (United States)

    Bulavin, Iliya

    type. Statocytes preserved their polarity in control but it was disturbed under clinorotation due to amyloplast distribution in the cytoplasm whole volume and/or their localization in the cell center. Structural rearrangements occurred similarly in statocytes under their transformation in secretory cells in control and under clinorotation. A characteristic features of the root proper meristematic cells in the control and in the experiment are central nucleus location, the great diversity of a size and a shape of mitochondria and plastids, poorly ER development, the presence of some small ER-bodies. As cells passed in the DEZ, their size enlarged but a nucleus can preserve the central location. A quantity of ER-cistern, vacuoles, and ER-bodies increased also. Dictyosomes acquired polarity and produced many Golgi vesicles. In CEZ cells, a large vacuole occupied the cell center, and the cytoplasm with organelles was on the cell periphery. So, we can conclude that under clinorotation: 1) the structure of a cap and growth zones of A. thaliana wild type and scr mutant roots formed de novo in vitro as similar to that in control; 2) a gaviperceptive apparatus formed in both objects but did not function. The obtained data allow to propose the model “Rhizogenesis in vitro” for using in spaceflight experiments to study the influence of real microgravity on the cellular differentiation and basic processes.

  20. Anatomy, histochemistry and ultrastructure of seed and somatic embryo of Acrocomia aculeata (Arecaceae Anatomia, histoquímica e ultra-estrutura da semente e do embrião somático de Acrocomia aculeata (Arecaceae

    Directory of Open Access Journals (Sweden)

    Elisa Ferreira Moura


    Full Text Available Macaw palm (Acrocomia aculeata (Jacq. Lodd. ex Mart. is a tropical species with multiple uses, including oil supply for biodiesel production. However, structural and physiological studies of the seed are still scarce, as well as in vitro propagation techniques. The aim of this study was to characterize the anatomy, histochemistry and ultrastructure of the seed and the in vitro somatic embryo of A. aculeata, gaining insight into relationships between the post-harvest seed behavior and the somatic embryo conversion to plant. The zygotic embryo and the endosperm show high quantities of protein and lipids, stored in protein and lipid bodies, respectively. The cell wall storage polysaccharides occur in the thickened cell wall of the endosperm. The absence of vacuoles and the few organelles, except for the storage ones, indicate low metabolic state of the zygotic embryo, which suggests a orthodox behavior of the seed at maturity. The somatic embryo shows a shoot meristem with few leaf primordia, vacuolated cells and, occasionally, amyloplasts , but not lipid or protein reserves. These characteristics, common in somatic embryogenesis, could be associated with the maturation phase, and, consequently, with the low conversion of these embryos into plants.A macaúba (Acrocomia aculeata (Jacq. Lodd. ex Mart. é uma espécie tropical com múltiplos usos, incluindo a geração de óleo para a produção de biodiesel. Entretanto, estudos estruturais e fisiológicos da semente ainda são escassos, assim como as técnicas de propagação in vitro. Caracterizaram-se a anatomia, a histoquímica e ultraestrutura da semente e do embrião somático de A. aculeata obtido in vitro, visando a obter relações entre o comportamento pós-colheita da semente e a conversão em planta a partir do embrião somático. O embrião zigótico e o endosperma apresentam alta quantidade de proteínas e lipídios, estocados em corpos protéicos e lipídicos, respectivamente. Os

  1. 西瓜小孢子有丝分裂前后质体和线粒体的变化%Changes of Plastid and Mitochondria Before and After Microspore Division in Watermelon

    Institute of Scientific and Technical Information of China (English)



    To understand morphological changes of plastids and mitochondria before and after microspore division as well as the distribution of these organelles to vegetative and generative cells during microspore division, microspores at different developmental stages and newly born 2-celled pollen grains were investigated by means of transmission electron microscopy. Plastids in microspores at all developmental stages do not have internal membranous structures, and do not accumulate starch or any other forms of storage reserves, indicating an undifferentiated status. In contrast, plastids in the newly born 2-celled pollen grain accumulate a huge amount of starch, indicating that the microspore division stimulates differentiation of proplastids into amyloplasts. In the newly born 2-celled pollen grain, the generative cell does not contain plastids, while the newly born vegetative cell contains a large number of plastids, indicating a complete exclusion of plastids from the generative cel! during microspore mitotic division, i.e., watermelon possesses the mode of purely maternal plastid inheritance. Based on cytological observations, watermelon should be added to the list of Lycopersicon type. Mitochondria in microspores reveal few cristae, whereas cristae dramatically increase in number in 2-celled pollen grains.%为揭示西瓜小孢子有丝分裂前后质体和线粒体的变化以及它们向生殖细胞和营养细胞分配的特点,用透射电子显微技术对西瓜不同发育阶段小孢子及早期二细胞花粉的质体和线粒体进行比较研究。结果表明,小孢子的质体结构简单,不积累淀粉等贮藏物质,为原质体;二细胞花粉的质体不含内部膜系统,但积累大量淀粉,表明小孢子有丝分裂引起原质体向造粉体分化。二细胞花粉的质体都在营养细胞中,生殖细胞不含质体,显然,小孢子分裂过程中质体只分配给营养细胞而不分配给生殖细胞,这一不均等

  2. Pharmacobotanic characterization of young stems and stem barks of Rauvolfia sellowii Müll. Arg., Apocynaceae

    Directory of Open Access Journals (Sweden)

    Leopoldo Clemente Baratto


    Full Text Available Rauvolfia sellowii Müll. Arg. (Apocynaceae, a Brazilian native tree rich in indole alkaloids, is known as "pau-pra-tudo" and popularly used as hypocholesterolemic, hypoglycemic and antihypertensive. The aim of this work was to study the anatomy of the young stems and stem barks of this medicinal plant, in order to contribute to the identification of the species as a drug. The plant material was fixed and prepared according to standard microtechniques. The young stems have remaining epidermis, but a suberified peridermis is evident. The phellogen is located in the cortical region, forming suber externally. Underneath the phellogen, lies the phelloderm and collenchymatic region. In the cortex, there are numerous laticifers and some fibers. There is an incomplete sclerenchymatic sheath, consisting of several groups of fibers and stone cells. The stem has internal phloem ordered as isolated groups side by side. Numerous laticifers, calcium oxalate crystals, idioblasts and amyloplasts are found in the cortex, phloem, xylem and pith. The stem bark has many layers of suber and cortical parenchyma, a sheath composed of fibers and stone cells totally lignified, and external phloem. These anatomical characteristic, taken together, can be used as quality control parameters for this species.Rauvolfia sellowii Müll. Arg. (Apocynaceae, uma árvore nativa brasileira rica em alcaloides indólicos, é conhecida como "pau-pra-tudo" e utilizada popularmente como hipocolesterolêmica, hipoglicêmica e anti-hipertensiva. O presente trabalho teve como objetivo caracterizar anatomicamente o caule jovem e a casca caulinar dessa planta medicinal, a fim de contribuir para a identificação e autenticidade da droga. O material vegetal foi fixado e submetido às microtécnicas usuais. O caule jovem possui epiderme remanescente, porém uma periderme suberificada é observada. O felogênio instala-se na região cortical, formando súber externamente. Subjacentes ao felog

  3. Preparation and identification of water-soluble calcium-binding protein from grape (Vitis vinifera L.) seeds%葡萄籽中水溶性钙结合蛋白的分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    吕晨艳; 赵广华


    Calcium is an essential nutrient required for critical biological functions such as nerve conduction, muscle contraction, mitosis, blood coagulation, and structural support of the skeleton.Dietary calcium intake is of general interest for human beings, particularly for infants and young children, when growth is accelerated. Milk and milk products as effective calcium supplements are generally accepted by human race with their high bioavailability. However, less consumption of milk in industrialized countries leads to inadequate calcium intake. Therefore, it is important to explore an alternate source for calcium supplement. On the other hand, dried grape seeds are likewise rich in lipids (22.07%), carbohydrates (12.51%) and proteins (11.94%) (w/w) and grape seeds as by-product during juice production can be an alternative source of protein. Meanwhile, this study demonstrates that grape seeds are rich in calcium ((5.62±0.01) g/kg for embryonic cells and (6.32± 0.01) g/kg for intact grape seeds), which was identified by ICP-AES. The calcium was mainly distributed in the stroma of the amyloplasts and around the starch granules, which was observed under TEM (Transmission Electron Microscope). Further study indicates that water-soluble protein from grape (Vitis viniferaL.) seeds (WSPG) contained two major components, one of which was 11S globulin-like protein mainly responsible for the binding of calcium in WSPG and the other was a novel protein (Protein A). The calcium contents of protein isolate from each step were identified by ICP-AES as well. When a traditional alkali extraction and acid precipitation method was used for isolation of WSPG, many binding calcium ions were lost. It is worth noting that the protein composition of grape seed protein obtained by both 30%-50% (NH4)2SO4 sediment and the alkali extraction and acid precipitation method was nearly identical, which consisted of protein A and protein B at a ratio of 2 to 3, but the content of calcium in the

  4. Structure and Development of Endosperm Transfer Cells in Wheat%小麦胚乳传递细胞发育的结构观察

    Institute of Scientific and Technical Information of China (English)

    王慧慧; 王峰; 刘大同; 顾蕴洁; 王忠


    In developing wheat caryopsis,grain filling nutrients transport into the endosperm via endosperm transfer cells(ETCs).The developmental extent of ETCs is in close relation to yield and qualities of wheat.In this paper we investigated the developmental process and structural features of ETCs in wheat systematically by the combination of light microscopy and transmission electron microscopy.Main results were as follows:(1) ETCs occured in the endosperm epidermis bordering the endosperm cavity and were the first cell type histologically differentiated during endosperm development.ETCs could be divided into two subtypes: outer 1~2 layer of aleurone transfer cells and inner 1~2 layer of starchy endosperm transfer cells.(2) As caryopsis matured,the nuclei of starchy endosperm transfer cells disappeared,while the nuclei of aleurone transfer cells remained intact.(3) The development of ETCs showed obvious polarity and the pattern of temporal and spatial gradient.(4) Aleurone transfer cells had dense cytoplasm rich in rough endoplasmic reticulum,mitochondria,dictyosomes and lipid bodies;their plasma membrane wrinkled and in some region evaginated to form numerous plasmatubules.(5) Starchy endosperm transfer cells had sparse cytoplasm rich in amyloplasts and were highly vacuolated.(6) Lots of plasmodesmata penetrated into adjoining primary walls of ETCs where thickening and wall ingrowths didn't occur.(7) Mitochondria in ETCs showed polarized distribution,and most of them lied in the proximity of plasma membrane.The structural features of ETCs suggested that they may play a role in nutrients transportation into the endosperm both via apoplastic and symplastic pathway.%为从细胞学方面了解小麦产量和品质的形成机制,以扬麦5号为材料,利用光镜和透射电镜观察了小麦颖果发育过程中胚乳传递细胞的结构变化,并探讨了胚乳传递细胞的生理功能。结果表明:(1)胚乳传递细胞

  5. 银杏雌配子体发育过程中淀粉和蛋白质的积累与代谢%Research on Starch and Protein Accumulation and Metabolism During the Development of the Ginkgo biloba Female Gametophyte

    Institute of Scientific and Technical Information of China (English)

    陆彦; 王莉; 潘烨; 陈鹏; 王頔; 谢燕; 金鑫鑫


    tentpole strongly increased. Nutrients in the endosperm cells around the archegonia were drastically reduced, and the endosperm cells and neck cells began to disintegrate. Jacket cells, tentpole, neck cells, and the endosperm tissue around archegonia all disintegrated after fertilization. We also found that the accumulation of nutrients in the endosperm underwent four stages: Parenchyma cell proliferation, the formation of nutrients, a rapid increase in nutrient accumulation, and a slight increase in nutrient accumulation. Finally, we found that starch grains developed from amyloplasts that contain one or more amyloid. These starch grains proliferated by budding or by constriction in the middle. Proteins present included P1 in the inner endosperm cell, and P2 in the outer aleurone layer. Before fertilization, the jacket cells and tentpole were highly active, and filled with high amounts of starch and protein, as well as mitochondria, endoplasmic retieulum and vesicles.