Variation in stem morphology and movement of amyloplasts in white spruce grown in the weightless environment of the International Space Station.

PubMed

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

2015-01-01

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. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

Wortmannin-induced vacuole fusion enhances amyloplast dynamics in Arabidopsis zigzag1 hypocotyls

PubMed Central

Alvarez, Ashley Ann; Han, Sang Won; Toyota, Masatsugu; Brillada, Carla; Zheng, Jiameng; Gilroy, Simon

2016-01-01

Gravitropism in Arabidopsis shoots depends on the sedimentation of amyloplasts in the endodermis, and a complex interplay between the vacuole and F-actin. Gravity response is inhibited in zigzag-1 (zig-1), a mutant allele of VTI11, which encodes a SNARE protein involved in vacuole fusion. zig-1 seedlings have fragmented vacuoles that fuse after treatment with wortmannin, an inhibitor of phosphatidylinositol 3-kinase, and underscore a role of phosphoinositides in vacuole fusion. Using live-cell imaging with a vertical stage microscope, we determined that young endodermal cells below the apical hook that are smaller than 70 μm in length are the graviperceptive cells in dark-grown hypocotyls. This result was confirmed by local wortmannin application to the top of zig-1 hypocotyls, which enhanced shoot gravitropism in zig-1 mutants. Live-cell imaging of zig-1 hypocotyl endodermal cells indicated that amyloplasts are trapped between juxtaposed vacuoles and their movement is severely restricted. Wortmannin-induced fusion of vacuoles in zig-1 seedlings increased the formation of transvacuolar strands, enhanced amyloplast sedimentation and partially suppressed the agravitropic phenotype of zig-1 seedlings. Hypergravity conditions at 10 g were not sufficient to displace amyloplasts in zig-1, suggesting the existence of a physical tether between the vacuole and amyloplasts. Our results overall suggest that vacuole membrane remodeling may be involved in regulating the association of vacuoles and amyloplasts during graviperception. PMID:27816929

A morphometric analysis of the redistribution of organelles in columella cells of horizontally-oriented roots of Zea mays

NASA Technical Reports Server (NTRS)

Moore, R.

1986-01-01

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.

Mechanisms of graviperception and response in pea seedlings

NASA Technical Reports Server (NTRS)

Galston, A. W.

1984-01-01

A new method for the mass isolation and purification of multigranular amyloplasts from the bundle sheath parenchyma of etiolated pa epicotyls was presented. These bodies, which displace within 2+3 minutes of exposure to 1 x g, are probably the gravity receptors (statoliths) in this plant. These amyloplasts were characterized as having a doublemembrane with a surface-localized ATPase, a high calcium content, and their own genomic DNA. These amyloplasts are investigated as to (a) the reasons for their especially high density, probable related to their starch content, (b) the possible identity of their DNA with the DNA of chloroplasts and unigranular amyloplasts, and (c) possible importance of their high calcium content.

Plastid-to-Nucleus Retrograde Signals Are Essential for the Expression of Nuclear Starch Biosynthesis Genes during Amyloplast Differentiation in Tobacco BY-2 Cultured Cells1[W][OA

PubMed Central

Enami, Kazuhiko; Ozawa, Tomoki; Motohashi, Noriko; Nakamura, Masayuki; Tanaka, Kan; Hanaoka, Mitsumasa

2011-01-01

Amyloplasts, a subtype of plastid, are found in nonphotosynthetic tissues responsible for starch synthesis and storage. When tobacco (Nicotiana tabacum) Bright Yellow-2 cells are cultured in the presence of cytokinin instead of auxin, their plastids differentiate from proplastids to amyloplasts. In this program, it is well known that the expression of nucleus-encoded starch biosynthesis genes, such as ADP-Glucose Pyrophosphorylase (AgpS) and Granule-Bound Starch Synthase (GBSS), is specifically induced. In this study, we investigated the roles of plastid gene expression in amyloplast differentiation. Microarray analysis of plastid genes revealed that no specific transcripts were induced in amyloplasts. Nevertheless, amyloplast development accompanied with starch biosynthesis was drastically inhibited in the presence of plastid transcription/translation inhibitors. Surprisingly, the expression of nuclear AgpS and GBSS was significantly repressed by the addition of these inhibitors, suggesting that a plastid-derived signal(s) that reflects normal plastid gene expression was essential for nuclear gene expression. A series of experiments was performed to examine the effects of intermediates and inhibitors of tetrapyrrole biosynthesis, since some of the intermediates have been characterized as candidates for plastid-to-nucleus retrograde signals. Addition of levulinic acid, an inhibitor of tetrapyrrole biosynthesis, resulted in the up-regulation of nuclear AgpS and GBSS gene expression as well as starch accumulation, while the addition of heme showed opposite effects. Thus, these results indicate that plastid transcription and/or translation are required for normal amyloplast differentiation, regulating the expression of specific nuclear genes by unknown signaling mechanisms that can be partly mediated by tetrapyrrole intermediates. PMID:21771917

The structure of the stem endodermis in etiolated pea seedlings

NASA Technical Reports Server (NTRS)

Sack, F. D.

1987-01-01

Differentiation of the endodermis was examined in third internodes of etiolated Pisum sativum L. cv. Alaska seedlings. The endodermis in young internodes contains large, sedimented amyloplasts; in older internodes, a casparian strip differentiates and the endodermis becomes depleted of starch except for the proximal region of the stem, which retains sedimented amyloplasts and remains graviresponsive. Sedimentation occurs in the hook but does not occur consistently until cells reach the base of the hook, where the axis becomes vertical, rapid cell elongation starts, and amyloplast diameter increases substantially. Contact between endoplasmic reticulum and amyloplasts was observed. Endoplasmic reticulum is not distributed polarly with respect to gravity. No symplastic or apoplastic blockages exist in the endodermis at the level of the stem where lateral gradients may be established during tropic curvature.

Caulonemal gravitropism and amyloplast sedimentation in the moss Funaria

NASA Technical Reports Server (NTRS)

Schwuchow, J. M.; Kim, D.; Sack, F. D.

1995-01-01

Caulonemata of the moss Funaria were examined to determine whether they are gravitropic. Funaria and Physcomitrella were also evaluated to compare amyloplast sedimentation with that of Ceratodon. Protonemata were either chemically fixed in place or examined alive using infrared timelapse videomicroscopy. Funaria caulonemata were found to be negatively gravitropic, i.e., they grew upwards in the dark. Upward curvature reversed temporarily before cytokinesis in Funaria, a phenomenon already known for Ceratodon and Physcomitrella. Most horizontal and upward-curving Funaria tip cells contained a broad subapical zone where plastid sedimentation occurred. In dark-grown Physcomitrella caulonemata, sedimentation was detected by the presence of a thin, amyloplast-free strip of cytoplasm at the top of the cell. These results suggest that gravitropism and subapical amyloplast sedimentation may be relatively common in moss caulonemata.

Organelle sedimentation in gravitropic roots of Limnobium is restricted to the elongation zone

NASA Technical Reports Server (NTRS)

Sack, F. D.; Kim, D.; Stein, B.

1994-01-01

Roots of the aquatic angiosperm Limnobium spongia (Bosc) Steud. were evaluated by light and electron microscopy to determine the distribution of organelle sedimentation towards gravity. Roots of Limnobium are strongly gravitropic. The rootcap consists of only two layers of cells. Although small amyloplasts are present in the central cap cells, no sedimentation of any organelle, including amyloplasts, was found. In contrast, both amyloplasts and nuclei sediment consistently and completely in cells of the elongation zone. Sedimentation occurs in one cell layer of the cortex just outside the endodermis. Sedimentation of both amyloplasts and nuclei begins in cells that are in their initial stages of elongation and persists at least to the level of the root where root hairs emerge. This is the first modern report of the presence of sedimentation away from, but not in, the rootcap. It shows that sedimentation in the rootcap is not necessary for gravitropic sensing in at least one angiosperm. If amyloplast sedimentation is responsible for gravitropic sensing, then the site of sensing in Limnobium roots is the elongation zone and not the rootcap. These data do not necessarily conflict with the hypothesis that sensing occurs in the cap in other roots, since Limnobium roots are exceptional in rootcap origin and structure, as well as in the distribution of organelle sedimentation. Similarly, if nuclear sedimentation is involved in gravitropic sensing, then nuclear mass would function in addition to, not instead of, that of amyloplasts.

Localization of cells containing sedimented amyloplasts in the shoots of normal and lazy rice seedlings.

PubMed

Abe, K; Takahashi, H; Suge, H

1994-12-01

We have examined the localization of the cells containing sedimented amyloplasts (putative statocytes) and its relation to the graviresponding sites in the shoots of normal and lazy rice seedlings. All graviresponsive organs of the shoots of normal rice seedlings, the mesocotyl, the coleoptile and the leaf-sheath base, were found to possess the statocytes. This is the first indication that mesocotyl senses gravity by its own cells in inducing gravitropic bending in rice seedlings. In lazy-Kamenoo, although the shoots lost their gravitropic response with the advance of age, sedimentation of amyloplasts itself might not be attributable to the agravitropic growth of the shoots, because, including those of the leaf-sheath bases that had lost their response to gravity, sedimented amyloplasts appeared to be identical to those of normal Kamenoo and of younger seedlings of lazy-Kamenoo whose gravitropism is still apparent.

Pea amyloplast DNA is qualitatively similar to pea chloroplast DNA

NASA Technical Reports Server (NTRS)

Gaynor, J. J.

1984-01-01

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.

Genetic analysis of the role of amyloplasts in shoot gravisensing

NASA Astrophysics Data System (ADS)

Tasaka, M.; Morita, M.

Plant can change the growth direction after sensing the gravity orientation This response calls gravitropism and the initial step is the gravisensing We have isolated many Arabidopsis mutants shoot gravitropism sgr with reduced or no gravitropic response in inflorescence stems The analysis of sgr1 and sgr7 revealed that endoderm cells in the inflorescence stems were gravisensing sites zig zigzag sgr4 and sgr3 showed no or reduced gravitropism in shoot respectively and their amyloplasts thought to be statoliths did not sedimented to the orientation of gravity in the endoderm cells ZIG encoded a SNARE AtVTI11 and SGR3 encoded other SNARE AtVAM3 These two SNAREs made a complex in the shoot endoderm cells suggesting that the vesicle transport from trans-Golgi network TGN to prevacuolar compartment PVC and or vacuole was involved in the amyloplasts localization and movement The analysis to visualize amyloplasts and vacuolar membrane in living endoderm cells supported that the vacuole function was important for the amyloplasts movement Recently we have isolated many suppressor mutants of zig One of them named zig suppressor zip 1 had a point mutation in the gene encoded other SNARE of AtVTI12 This protein is a homologous to ZIG AtVTI11 and these two proteins have partially redundant functions Although wild type At VTI 12 could not rescued zig mutated AtVTI12 protein ZIP1 could almost completely play the part of ZIG In zigzip1 amyloplasts in endoderm cells sedimented normally and the shoots showed normal gravitropic response The other

Mechanotransduction molecules in the plant gravisensory response: amyloplast/statolith membranes contain a beta 1 integrin-like protein

NASA Technical Reports Server (NTRS)

Lynch, T. M.; Lintilhac, P. M.; Domozych, D.

1998-01-01

It has been hypothesized that the sedimentation of amyloplasts within root cap cells is the primary event in the plant gravisensory-signal transduction cascade. Statolith sedimentation, with its ability to generate weighty mechanical signals, is a legitimate means for organisms to discriminate the direction of the gravity vector. However, it has been demonstrated that starchless mutants with reduced statolith densities maintain some ability to sense gravity, calling into question the statolith sedimentation hypothesis. Here we report on the presence of a beta 1 integrin-like protein localized inside amyloplasts of tobacco NT-1 suspension culture, callus cells, and whole-root caps. Two different antibodies to the beta 1 integrin, one to the cytoplasmic domain and one to the extracellular domain, localize in the vicinity of the starch grains within amyloplasts of NT-1. Biochemical data reveals a 110-kDa protein immunoprecipitated from membrane fractions of NT-1 suspension culture indicating size homology to known beta 1 integrin in animals. This study provides the first direct evidence for the possibility of integrin-mediated signal transduction in the perception of gravity by higher plants. An integrin-mediated pathway, initiated by starch grain sedimentation within the amyloplast, may provide the signal amplification necessary to explain the gravitropic response in starch-depleted cultivars.

Deficiency of Starch Synthase IIIa and IVb Alters Starch Granule Morphology from Polyhedral to Spherical in Rice Endosperm1

PubMed Central

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

2016-01-01

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

Hydrotropism Interacts with Gravitropism by Degrading Amyloplasts in Seedling Roots of Arabidopsis and Radish1

PubMed Central

Takahashi, Nobuyuki; Yamazaki, Yutaka; Kobayashi, Akie; Higashitani, Atsushi; Takahashi, Hideyuki

2003-01-01

In response to a moisture gradient, roots exhibit hydrotropism to control the orientation of their growth. To exhibit hydrotropism, however, they must overcome the gravitropism that is dominant on Earth. We found that moisture gradient or water stress caused immediate degradation of the starch anchors, amyloplasts, in root columella cells of Arabidopsis and radish (Raphanus sativus). Namely, development of hydrotropic response was accompanied by a simultaneous reduction in starch content in columella cells. Rapid degradation of amyloplasts in columella cells also occurred in the water-stressed roots with sorbitol or mannitol. Both hydrotropically stimulated and water-stressed roots showed a reduced responsiveness to gravity. Roots of a starchless mutant, pgm1-1, showed an enhanced hydrotropism compared with that of the wild type. These results suggest that the reduced responsiveness to gravity is, at least in part, attributable to the degradation of amyloplasts in columella cells. Thus, the reduction in gravitropism allows the roots to exhibit hydrotropism. PMID:12805610

Root negative gravitropism is accompanied with displacing of columella amyloplasts to the statocyte upper longitudinal cell wall

NASA Astrophysics Data System (ADS)

Kordyum, Elizabeth; Sobol, Margaryta; Kalinina, Yana; Bogatina, Nina; Kondrachuk, Alexander

Recently it was shown that roots reveal negative gravitropism in the weak combined magnetic field (CMF) with the frequency resonance to the cyclotron frequency of Ca2+ ions. A negative gravitropic reaction in the CMF occurs by a usual physiological process. Experiments in the CMF confirmed that gravitropism is plastid-based and Ca2+ ions participate in this process. Unlike control, amyloplasts-statoliths are not displacing on the lower side of a gravistimulated root but tend to group in the center of a statocyte during 30 min under gravistimulation in the CMF. In an hour of gravistimulation, they are localized near one of the statocyte longitudinal wall. Now we determined that amyloplasts are localized along the statocyte upper longitudinal side. It is of a special interest that a root is bending to the same direction with displacing of amyloplasts: in positive gravitropism - downwards, in negative gravitropism - upwards. On the basis of the obtained data there is a question, what forces promote displacing of amyloplasts against a gravitational vector? In the paper, three possible explanations are discussed: 1) CMF + Ca2+ action on the distribution of elastic forces in cytoskeleton, 2) CMF + Ca2+ action on the distribution of electric field in statocytes, and 3) CMF action on energy and direction of Ca2+ ion rotation according to the ion cyclotron resonance model that can lead to paradoxical Ca2+ redistribution.

Amyloplast Distribution Directs a Root Gravitropic Reaction

NASA Astrophysics Data System (ADS)

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 with regard to the participation of calcium ions and cytoskeletal elements in these processes is therefore substantial but still circumstantial and requires new experimental data. Using a new model - weak combined magnetic fields (CMFs), which elicit a variety of responses in plants, growth rate and fresh weight, seed germination, Ca2+ concentration, membrane permeability, with a frequency resonance to cyclotron frequency of calcium ions, we firstly showed that a root positive gravitropic reaction changes on a negative one. In this case, the paradoxical displacement of amylopasts-statoliths to the upper longitudinal cell wall of statocytes occurred in the direction opposite to a gravitational vector. Displacement of amyloplasts, which contain the abundance of free Ca2+ in the stroma, was accompanied with Ca2+ redistribution in the same direction in the cytosol and increasing around amyloplasts in comparison with the state magnetic field. In the elongation zone, calcium ions accumulated in the upper site of a gravistimulated root unlike a positive gravitropic reaction, and a root is bending in the same direction in which amyloplasts are displacing. It seems that a root gravitropic reaction, if it began, occurs by an usual physiological way resulting in root bending with an opposite sign. It is of a special interest that a root is bending to the same direction with displacing of amyloplasts: in positive gravitropism - downwards, in negative gravitropism - upwards. Peculiarities of calcium ion redistribution in statocytes under gravistimulation in such combined magnetic field are a new additional evidence of a Ca2+ ion significant role in gravitropism. Thus, our data support the starch-statolith hypothesis but also pose the question as to which forces displace amyloplasts against the gravity vector? We hope that these data will stimulate new research to better understand the mechanisms of plant graviperception and graviresponse. Gravistimulation of a root in the CMF with the frequency resonance to the cyclotron frequency of Ca2+ ions is an effective model for future research of the mechanism of plant gravitropism, including a Ca2+ role in plant physiological growth reactions.

The change of amyloplasts structure and composition of storage starch in potato minitubers during imitated microgravity

NASA Astrophysics Data System (ADS)

Nedukha, O. M.; Kordyum, E. L.; Martyn, G. M.; Schnyukova, E. I.

Potato was designated for food production in the controlled ecological life-support system CELSS because its tubers as it is known contain starch and significant protein content and are edible food after the long-term storage We used the cultivation of potato miniplants under influence of long-term horizontal clinorotation 2 rev min which imitated microgravity as a model for the technology of potato food production in the CELSS The aim of our work was to determine content and composition storage starch as well as amyloplast ultrastructure of storage parenchyma cells in potato minitubers formed under long-term to 6 weeks slow horizontal clinorotation 2 rpm Minitubers developed from axillary buds of potato miniplants growing in the aseptic stationary conditions and under clinorotation Methods of scanning and transmission electron microscopy were used for the study of surface and ultrastructure of amyloplasts the biochemical method by Hovenkamp-Hermelink et al 1988 - for study of starch composition Some differences were observed in amyloplast structure under clinorotation namely increased volume of starch grains in plastid decreased stroma volume changed structure of envelope membranes in comparison with the stationary control Besides an appearance of fraction of gigantic amyloplasts in central layers of parenchyma was observed under clinorotation after 4 weeks of growth The total starch content increased and reached to 219 5 - 4 1 mg g FW at 6 weeks of clinorotation it was 167 5 - 5 6 mg g FW in the control minitubers A ratio of

The Effect of Weak Combined Magnetic Field on Root Gravitropism and a Role of Ca2+ Ions Therein

NASA Astrophysics Data System (ADS)

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

Ultrastructure of pea and cress root statocytes exposed to high gradient magnetic field

NASA Astrophysics Data System (ADS)

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 appearance of significant amounts of electron-dense granules in the cytoplasm and the nucleus (particularly, in the nucleolus); the effect was most evident in HGMF. Testing the chemical composition of such deposits is going on. The data presented statocyte responses indicate similarity the effects of magneto- and gravistimulation at the ultrastructural level. Thus, the root curvature in HGMF is the plant response to displacement of amyloplasts by ponderomotive force, which can serve as tool for investigation of graviperception mechanism and can provide directional stimulus for plant growth in microgravity. (Financial support by STCU: NN-13R).

Directional gravity sensing in gravitropism.

PubMed

Morita, Miyo Terao

2010-01-01

Plants can reorient their growth direction by sensing organ tilt relative to the direction of gravity. With respect to gravity sensing in gravitropism, the classic starch statolith hypothesis, i.e., that starch-accumulating amyloplast movement along the gravity vector within gravity-sensing cells (statocytes) is the probable trigger of subsequent intracellular signaling, is widely accepted. Several lines of experimental evidence have demonstrated that starch is important but not essential for gravity sensing and have suggested that it is reasonable to regard plastids (containers of starch) as statoliths. Although the word statolith means sedimented stone, actual amyloplasts are not static but instead possess dynamic movement. Recent studies combining genetic and cell biological approaches, using Arabidopsis thaliana, have demonstrated that amyloplast movement is an intricate process involving vacuolar membrane structures and the actin cytoskeleton. This review covers current knowledge regarding gravity sensing, particularly gravity susception, and the factors modulating the function of amyloplasts for sensing the directional change of gravity. Specific emphasis is made on the remarkable differences in the cytological properties, developmental origins, tissue locations, and response of statocytes between root and shoot systems. Such an approach reveals a common theme in directional gravity-sensing mechanisms in these two disparate organs.

Changes in the distribution of plastids and endoplasmic reticulum during cellular differentiation in root caps of Zea mays

NASA Technical Reports Server (NTRS)

Moore, R.; McClelen, C. E.

1985-01-01

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.

FLOURY ENDOSPERM7 encodes a regulator of starch synthesis and amyloplast development essential for peripheral endosperm development in rice

PubMed Central

Zhang, Long; Ren, Yulong; Lu, Bingyue; Yang, Chunyan; Feng, Zhiming; Liu, Zhou; Chen, Jun; Ma, Weiwei; Wang, Ying; Yu, Xiaowen; Wang, Yunlong; Zhang, Wenwei; Wang, Yihua; Liu, Shijia; Wu, Fuqing; Zhang, Xin; Guo, Xiuping; Bao, Yiqun; Jiang, Ling; Wan, Jianmin

2016-01-01

In cereal crops, starch synthesis and storage depend mainly on a specialized class of plastids, termed amyloplasts. Despite the importance of starch, the molecular machinery regulating starch synthesis and amyloplast development remains largely unknown. Here, we report the characterization of the rice (Oryza sativa) floury endosperm7 (flo7) mutant, which develops a floury-white endosperm only in the periphery and not in the inner portion. Consistent with the phenotypic alternation in flo7 endosperm, the flo7 mutant had reduced amylose content and seriously disrupted amylopectin structure only in the peripheral endosperm. Notably, flo7 peripheral endosperm cells showed obvious defects in compound starch grain development. Map-based cloning of FLO7 revealed that it encodes a protein of unknown function. FLO7 harbors an N-terminal transit peptide capable of targeting functional FLO7 fused to green fluorescent protein to amyloplast stroma in developing endosperm cells, and a domain of unknown function 1338 (DUF1338) that is highly conserved in green plants. Furthermore, our combined β-glucuronidase activity and RNA in situ hybridization assays showed that the FLO7 gene was expressed ubiquitously but exhibited a specific expression in the endosperm periphery. Moreover, a set of in vivo experiments demonstrated that the missing 32 aa in the flo7 mutant protein are essential for the stable accumulation of FLO7 in the endosperm. Together, our findings identify FLO7 as a unique plant regulator required for starch synthesis and amyloplast development within the peripheral endosperm and provide new insights into the spatial regulation of endosperm development in rice. PMID:26608643

Actin microfilaments in presumptive statocytes of root caps and coleoptiles

NASA Technical Reports Server (NTRS)

White, R. G.; Sack, F. D.

1990-01-01

Rhodamine-phalloidin was used to determine the distribution of actin microfilament bundles (mfb) in cells thought to be the site of gravity perception (statocytes) in coleoptiles and root caps of Zea mays and Hordeum vulgare. In coleoptile cells, amyloplasts were usually observed in close proximity to thick mfb, which often appeared to divide into finer mfb adjacent to individual amyloplasts. The nucleus in these cells was surrounded by an extensive network of mfb, which were connected to thicker transvacuolar mfb. Columella cells of the root cap contained an extensive reticulum of fine mfb throughout the protoplast, but lacked the much thicker mfb seen in coleoptile cells. The distribution and extent of mfb observed in fixed cells correlates with patterns of streaming and amyloplast movement seen in living cells. A possible role for actin mfb in the perception of gravity is discussed.

Influence of microgravity on root-cap regeneration and the structure of columella cells in Zea mays

NASA Technical Reports Server (NTRS)

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

1987-01-01

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.

Intracellular magnetophoresis of amyloplasts and induction of root curvature

NASA Technical Reports Server (NTRS)

Kuznetsov, O. A.; Hasenstein, K. H.

1996-01-01

High-gradient magnetic fields (HGMFs) were used to induce intracellular magnetophoresis of amyloplasts. The HGMFs were generated by placing a small ferromagnetic wedge into a uniform magnetic field or at the gap edge between two permanent magnets. In the vicinity of the tip of the wedge the dynamic factor of the magnetic field, delta(H2/2), was about 10(9) Oe2.cm-1, which subjected the amyloplasts to a force comparable to that of gravity. When roots of 2-d-old seedlings of flax (Linum usitatissimum L.) were positioned vertically and exposed to an HGMF, curvature away from the wedge was transient and lasted approximately 1 h. Average curvature obtained after placing magnets, wedge and seedlings on a 1-rpm clinostat for 2 h was 33 +/- 5 degrees. Roots of horizontally placed control seedlings without rotation curved about 47 +/- 4 degrees. The time course of curvature and changes in growth rate were similar for gravicurvature and for root curvature induced by HGMFs. Microscopy showed displacement of amyloplasts in vitro and in vivo. Studies with Arabidopsis thaliana (L.) Heynh. showed that the wild type responded to HGMFs but the starchless mutant TC7 did not. The data indicate that a magnetic force can be used to study the gravisensing and response system of roots.

Gravity-regulated formation of the peg in developing cucumber seedlings

NASA Technical Reports Server (NTRS)

Takahashi, H.; Scott, T. K.

1994-01-01

It has been proposed that peg formation in the vascular transition region (TR zone) between the hypocotyl and the root in Cucurbitaceae seedlings is a gravimorphogenetic phenomenon. Initiation of the peg became visible 36 h after imbibition when cucumber (Cucumis sativus L. cv. Burpee Hybrid II) seeds were germinated in a horizontal position at 24 degrees C in the dark. Simultaneously, sedimented amyloplasts (putative statoliths) were apparent in the sheath cells surrounding the vascular strands, and in the cortical cells immediately adjacent to them, in the TR zone. In contrast, the other cortical cells, some of which were destined to develop into the peg, contained amyloplasts which were not sedimented. These results suggest that the graviperception mechanism for peg formation may be like that of statoliths in shoot gravitropism. By 48 h following imbibition, the cells of the TR zone still had sedimented amyloplasts but had lost their sensitivity to gravity, possibly because of their maturation.

Phytochromes inhibit hypocotyl negative gravitropism by regulating the development of endodermal amyloplasts through phytochrome-interacting factors

PubMed Central

Kim, Keunhwa; Shin, Jieun; Lee, Sang-Hee; Kweon, Hee-Seok; Maloof, Julin N.; Choi, Giltsu

2011-01-01

Phytochromes are red and far-red light photoreceptors that regulate various aspects of plant development. One of the less-understood roles of phytochromes is the inhibition of hypocotyl negative gravitropism, which refers to the loss of hypocotyl gravitropism and resulting random growth direction in red or far-red light. This light response allows seedlings to curve toward blue light after emergence from the soil and enhances seedling establishment in the presence of mulch. Phytochromes inhibit hypocotyl negative gravitropism by inhibiting four phytochrome-interacting factors (PIF1, PIF3, PIF4, PIF5), as shown by hypocotyl agravitropism of dark-grown pif1 pif3 pif4 pif5 quadruple mutants. We show that phytochromes inhibit negative gravitropism by converting starch-filled gravity-sensing endodermal amyloplasts to other plastids with chloroplastic or etioplastic features in red or far-red light, whereas PIFs promote negative gravitropism by inhibiting the conversion of endodermal amyloplasts to etioplasts in the dark. By analyzing transgenic plants expressing PIF1 with an endodermis-specific SCARECROW promoter, we further show that endodermal PIF1 is sufficient to inhibit the conversion of endodermal amyloplasts to etioplasts and hypocotyl negative gravitropism of the pif quadruple mutant in the dark. Although the functions of phytochromes in gravitropism and chloroplast development are normally considered distinct, our results indicate that these two functions are closely related. PMID:21220341

Phytochromes inhibit hypocotyl negative gravitropism by regulating the development of endodermal amyloplasts through phytochrome-interacting factors.

PubMed

Kim, Keunhwa; Shin, Jieun; Lee, Sang-Hee; Kweon, Hee-Seok; Maloof, Julin N; Choi, Giltsu

2011-01-25

Phytochromes are red and far-red light photoreceptors that regulate various aspects of plant development. One of the less-understood roles of phytochromes is the inhibition of hypocotyl negative gravitropism, which refers to the loss of hypocotyl gravitropism and resulting random growth direction in red or far-red light. This light response allows seedlings to curve toward blue light after emergence from the soil and enhances seedling establishment in the presence of mulch. Phytochromes inhibit hypocotyl negative gravitropism by inhibiting four phytochrome-interacting factors (PIF1, PIF3, PIF4, PIF5), as shown by hypocotyl agravitropism of dark-grown pif1 pif3 pif4 pif5 quadruple mutants. We show that phytochromes inhibit negative gravitropism by converting starch-filled gravity-sensing endodermal amyloplasts to other plastids with chloroplastic or etioplastic features in red or far-red light, whereas PIFs promote negative gravitropism by inhibiting the conversion of endodermal amyloplasts to etioplasts in the dark. By analyzing transgenic plants expressing PIF1 with an endodermis-specific SCARECROW promoter, we further show that endodermal PIF1 is sufficient to inhibit the conversion of endodermal amyloplasts to etioplasts and hypocotyl negative gravitropism of the pif quadruple mutant in the dark. Although the functions of phytochromes in gravitropism and chloroplast development are normally considered distinct, our results indicate that these two functions are closely related.

endodermal-amyloplast less 1 is a novel allele of SHORT-ROOT

NASA Astrophysics Data System (ADS)

Morita, Miyo T.; Saito, Chieko; Nakano, Akihiko; Tasaka, Masao

Plants can sense the direction of gravity and change the growth orientation of their organs. Arabidopsis mutants have been isolated and characterized in order to elucidate the molecular mechanisms of gravitropism. endodermal-amyloplast less 1 ( eal1) is a unique mutant that completely lacks gravitropism in inflorescence stems and exhibits reduced gravitropism in hypocotyls, whereas its roots showed normal gravitropism. Previously, it was suggested that differentiation or development of amyloplasts in shoot statocytes (endodermal cells) is affected by the eal1 mutation. Here, we have identified EAL1 as a SHORT-ROOT ( SHR) allele based on map position. Three nucleotides in the SHR coding region were deleted in the eal1 mutant, resulting in the deletion of just one amino acid. The protein encoded by the novel allele of SHR appears to have retained its function as a transcription factor since the endodermal cell layer was formed both in roots and in shoots of eal1. SCARECROW (SCR) promoter activity monitored by reporter protein expression was significantly decreased in eal1, suggesting that the activity of SHR lacking one amino acid is reduced. In addition, transcription levels of SHOOT GRAVITROPISM 5 (SGR5), which is mainly expressed in the endodermis of inflorescence stems, was markedly decreased. Together with the presence of abnormal endodermal amyloplasts in eal1, these results strongly suggest that the endodermis observed in eal1 is not sufficiently differentiated to execute shoot gravitropism.

Curvature in Arabidopsis inflorescence stems is limited to the region of amyloplast displacement

NASA Technical Reports Server (NTRS)

Weise, S. E.; Kuznetsov, O. A.; Hasenstein, K. H.; Kiss, J. Z.

2000-01-01

Gravitropic sensing in stems and stem-like organs is hypothesized to occur in the endodermis. However, since the endodermis runs the entire length of the stem, the precise site of gravisensing has been difficult to define. In this investigation of gravisensitivity in inflorescence stems of Arabidopsis, we positioned stems in a high gradient magnetic field (HGMF) on a rotating clinostat. Approximately 40% of the young, wild-type (WT) inflorescences, for all positions tested, curved toward the HGMF in the vicinity of the stem exposed to the field. In contrast, when the wedge was placed in the basal region of older inflorescence stems, no curvature was observed. As a control, the HGMF was applied to a starchless mutant, and 5% of the stems curved toward the field. Microscopy of the endodermis in the WT showed amyloplast displacement in the vicinity of the HGMF. Additional structural studies demonstrated that the basal region of WT stems experienced amyloplast displacement and, therefore, suggest this region is capable of gravity perception. However, increased lignification likely prevented curvature in the basal region. The lack of apical curvature after basal amyloplast displacement indicates that gravity perception in the base is not transmitted to the apex. Thus, these results provide evidence that the signal (and thus, response) resulting from perception in Arabidopsis inflorescence stems is spatially restricted.

Effects of microgravity and clinorotation on stress ethylene production in two starchless mutants of Arabidopsis thaliana

NASA Technical Reports Server (NTRS)

Gallegos, Gregory L.; Hilaire, Emmanuel M.; Peterson, Barbara V.; Brown, Christopher S.; Guikema, James A.

1995-01-01

Starch filled plastids termed amyloplasts, contained within columella cells of the root caps of higher plant roots, are believed to play a statolith-like role in the gravitropic response of roots. Plants having amyloplasts containing less starch exhibit a corresponding reduction in gravitropic response. We have observed enhanced ethylene production by sweet clover (Melilotus alba L.) seedlings grown in the altered gravity condition of a slow rotating clinostat, and have suggested that this is a stress response resulting from continuous gravistimulation rather than as a result of the simulation of a microgravity condition. If so, we expect that plants deficient in starch accumulation in amyloplasts may produce less stress ethylene when grown on a clinostat. Therefore, we have grown Arabidopsis thaliana in the small, closed environment of the Fluid Processing Apparatus (FPA). In this preliminary report we compare stationary plants with clinorotated and those grown in microgravity aboard Discovery during the STS-63 flight in February 1995. In addition to wildtype, two mutants deficient in starch biosynthesis, mutants TC7 and TL25, which are, respectively, deficient in the activity of amyloplast phosphoglucomutase and ADP-glucose pyrophosphorylase, were grown for three days before being fixed within the FPA. Gas samples were aspirated from the growth chambers and carbon dioxide and ethylene concentations were measured using a gas chromatograph. The fixed tissue is currently undergoing further morphologic and microscopic characterization.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, Woo Taek; Franceschi, V.R.; Okita, T.W.

The subcellular localization of ADPglucose pyrophosphorylase, a key regulatory enzyme in starch biosynthesis, was determined in developing potato tuber cells by immunocytochemical localization techniques at the light microscopy level. Specific labeling of ADPglucose pyrophosphorylase by either immunofluorescence or immunogold followed by silver enhancement was detected only in the amyloplasts and indicates that this enzyme is located exclusively in the amyloplasts in developing potato tuber cells. Labeling occurred on the starch grains and, in some instances, specific labeling patterns were evident which may be related to sites active in starch deposition.

Multiple roles for membrane-associated protein trafficking and signaling in gravitropism

PubMed Central

Strohm, Allison K.; Baldwin, Katherine L.; Masson, Patrick H.

2012-01-01

Gravitropism is a process that allows plant organs to guide their growth relative to the gravity vector. It requires them to sense changes in their orientation and generate a biochemical signal that they transmit to the tissues that drive organ curvature. Trafficking between the plasma membrane and endosomal compartments is important for all of these phases of the gravitropic response. The sedimentation of starch-filled organelles called amyloplasts plays a key role in sensing reorientation, and vacuolar integrity is required for amyloplast sedimentation in shoots. Other proteins associated with the vesicle trafficking pathway contribute to early gravity signal transduction independently of amyloplast sedimentation in both roots and hypocotyls. Phosphatidylinositol signaling, which starts at the plasma membrane and later affects the localization of auxin efflux facilitators, is a likely second messenger in the signal transduction phase of gravitropism. Finally, membrane-localized auxin influx and efflux facilitators contribute to a differential auxin gradient across the gravistimulated organs, which directs root curvature. PMID:23248632

Multiple roles for membrane-associated protein trafficking and signaling in gravitropism.

PubMed

Strohm, Allison K; Baldwin, Katherine L; Masson, Patrick H

2012-01-01

Gravitropism is a process that allows plant organs to guide their growth relative to the gravity vector. It requires them to sense changes in their orientation and generate a biochemical signal that they transmit to the tissues that drive organ curvature. Trafficking between the plasma membrane and endosomal compartments is important for all of these phases of the gravitropic response. The sedimentation of starch-filled organelles called amyloplasts plays a key role in sensing reorientation, and vacuolar integrity is required for amyloplast sedimentation in shoots. Other proteins associated with the vesicle trafficking pathway contribute to early gravity signal transduction independently of amyloplast sedimentation in both roots and hypocotyls. Phosphatidylinositol signaling, which starts at the plasma membrane and later affects the localization of auxin efflux facilitators, is a likely second messenger in the signal transduction phase of gravitropism. Finally, membrane-localized auxin influx and efflux facilitators contribute to a differential auxin gradient across the gravistimulated organs, which directs root curvature.

Root graviresponsiveness and cellular differentiation in wild-type and a starchless mutant of Arabidopsis thaliana

NASA Technical Reports Server (NTRS)

Moore, R.

1989-01-01

Primary roots of a starchless mutant of Arabidopsis thaliana L. are strongly graviresponsive despite lacking amyloplasts in their columella cells. The ultrastructures of calyptrogen and peripheral cells in wild-type as compared to mutant seedlings are not significantly different. The largest difference in cellular differentiation in caps of mutant and wild-type roots is the relative volume of plastids in columella cells. Plastids occupy 12.3% of the volume of columella cells in wild-type seedlings, but only 3.69% of columella cells in mutant seedlings. These results indicate that: (1) amyloplasts and starch are not necessary for root graviresponsiveness; (2) the increase in relative volume of plastids that usually accompanies differentiation of columella cells is not necessary for root graviresponsiveness; and (3) the absence of starch and amyloplasts does not affect the structure of calyptrogen (i.e. meristematic) and secretory (i.e. peripheral) cells in root caps. These results are discussed relative to proposed models for root gravitropism.

Amyloplast movement and gravityperception in Arabidopsis endoderm

NASA Astrophysics Data System (ADS)

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.

The gravitropism defective 2 Mutants of Arabidopsis Are Deficient in a Protein Implicated in Endocytosis in Caenorhabditis elegans1[w

PubMed Central

Silady, Rebecca A.; Kato, Takehide; Lukowitz, Wolfgang; Sieber, Patrick; Tasaka, Masao; Somerville, Chris R.

2004-01-01

The gravitropism defective 2 (grv2) mutants of Arabidopsis show reduced shoot phototropism and gravitropism. Amyloplasts in the shoot endodermal cells of grv2 do not sediment to the same degree as in wild type. The GRV2 gene encodes a 277-kD polypeptide that is 42% similar to the Caenorhabditis elegans RME-8 protein, which is required for endocytosis. We hypothesize that a defect in endocytosis may affect both the initial gravity sensing via amyloplasts sedimentation and the subsequent more general tropic growth response. PMID:15466218

Interaction between hydrotropism and gravitropism in seedling roots

NASA Astrophysics Data System (ADS)

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 controlling the directional growth of seedling roots.

The role of Arabidopsis Actin-Related Protein 3 in amyloplast sedimentation and polar auxin transport in root gravitropism

PubMed Central

Zou, Jun-Jie; Zheng, Zhong-Yu; Xue, Shan; Li, Han-Hai; Wang, Yu-Ren; Le, Jie

2016-01-01

Gravitropism is vital for shaping directional plant growth in response to the forces of gravity. Signals perceived in the gravity-sensing cells can be converted into biochemical signals and transmitted. Sedimentation of amyloplasts in the columella cells triggers asymmetric auxin redistribution in root tips, leading to downward root growth. The actin cytoskeleton is thought to play an important role in root gravitropism, although the molecular mechanism has not been resolved. DISTORTED1 (DIS1) encodes the ARP3 subunit of the Arabidopsis Actin-Related Protein 2/3 (ARP2/3) complex, and the ARP3/DIS1 mutant dis1-1 showed delayed root curvature after gravity stimulation. Microrheological analysis revealed that the high apparent viscosity within dis1-1 central columella cells is closely associated with abnormal movement trajectories of amyloplasts. Analysis using a sensitive auxin input reporter DII-VENUS showed that asymmetric auxin redistribution was reduced in the root tips of dis1-1, and the actin-disrupting drug Latrunculin B increased the asymmetric auxin redistribution. An uptake assay using the membrane-selective dye FM4-64 indicated that endocytosis was decelerated in dis1-1 root epidermal cells. Treatment and wash-out with Brefeldin A, which inhibits protein transport from the endoplasmic reticulum to the Golgi apparatus, showed that cycling of the auxin-transporter PIN-FORMED (PIN) proteins to the plasma membrane was also suppressed in dis1-1 roots. The results reveal that ARP3/DIS1 acts in root gravitropism by affecting amyloplast sedimentation and PIN-mediated polar auxin transport through regulation of PIN protein trafficking. PMID:27473572

The role of Arabidopsis Actin-Related Protein 3 in amyloplast sedimentation and polar auxin transport in root gravitropism.

PubMed

Zou, Jun-Jie; Zheng, Zhong-Yu; Xue, Shan; Li, Han-Hai; Wang, Yu-Ren; Le, Jie

2016-10-01

Gravitropism is vital for shaping directional plant growth in response to the forces of gravity. Signals perceived in the gravity-sensing cells can be converted into biochemical signals and transmitted. Sedimentation of amyloplasts in the columella cells triggers asymmetric auxin redistribution in root tips, leading to downward root growth. The actin cytoskeleton is thought to play an important role in root gravitropism, although the molecular mechanism has not been resolved. DISTORTED1 (DIS1) encodes the ARP3 subunit of the Arabidopsis Actin-Related Protein 2/3 (ARP2/3) complex, and the ARP3/DIS1 mutant dis1-1 showed delayed root curvature after gravity stimulation. Microrheological analysis revealed that the high apparent viscosity within dis1-1 central columella cells is closely associated with abnormal movement trajectories of amyloplasts. Analysis using a sensitive auxin input reporter DII-VENUS showed that asymmetric auxin redistribution was reduced in the root tips of dis1-1, and the actin-disrupting drug Latrunculin B increased the asymmetric auxin redistribution. An uptake assay using the membrane-selective dye FM4-64 indicated that endocytosis was decelerated in dis1-1 root epidermal cells. Treatment and wash-out with Brefeldin A, which inhibits protein transport from the endoplasmic reticulum to the Golgi apparatus, showed that cycling of the auxin-transporter PIN-FORMED (PIN) proteins to the plasma membrane was also suppressed in dis1-1 roots. The results reveal that ARP3/DIS1 acts in root gravitropism by affecting amyloplast sedimentation and PIN-mediated polar auxin transport through regulation of PIN protein trafficking. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Gravitropic mechanisms derived from space experiments and magnetic gradients.

NASA Astrophysics Data System (ADS)

Hasenstein, Karl H.; Park, Myoung Ryoul

2016-07-01

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 and different responses to microgravity conditions, induced curvature, ground controls, clinorotation, and magnetic field exposure.

Amyloplast Membrane Protein SUBSTANDARD STARCH GRAIN6 Controls Starch Grain Size in Rice Endosperm1

PubMed Central

Matsushima, Ryo; Maekawa, Masahiko; Kusano, Miyako; Tomita, Katsura; Kondo, Hideki; Nishimura, Hideki; Crofts, Naoko; Fujita, Naoko; Sakamoto, Wataru

2016-01-01

Starch is a biologically and commercially important polymer of glucose. Starch is organized into starch grains (SGs) inside amyloplasts. The SG size differs depending on the plant species and is one of the most important factors for industrial applications of starch. There is limited information on genetic factors regulating SG sizes. In this study, we report the rice (Oryza sativa) mutant substandard starch grain6 (ssg6), which develops enlarged SGs in endosperm. Enlarged SGs are observed starting at 3 d after flowering. During endosperm development, a number of smaller SGs appear and coexist with enlarged SGs in the same cells. The ssg6 mutation also affects SG morphologies in pollen. The SSG6 gene was identified by map-based cloning and microarray analysis. SSG6 encodes a protein homologous to aminotransferase. SSG6 differs from other rice homologs in that it has a transmembrane domain. SSG6-green fluorescent protein is localized in the amyloplast membrane surrounding SGs in rice endosperm, pollen, and pericarp. The results of this study suggest that SSG6 is a novel protein that controls SG size. SSG6 will be a useful molecular tool for future starch breeding and applications. PMID:26792122

Gravisensing in roots

NASA Astrophysics Data System (ADS)

Perbal, G.

1999-01-01

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.

Magnetophoretic induction of curvature in coleoptiles and hypocotyls

NASA Technical Reports Server (NTRS)

Kuznetsov, O. A.; Hasenstein, K. H.

1997-01-01

Coleoptiles of barley (Hordeum vulgare) were positioned in a high gradient magnetic field (HGMF, dynamic factor gradient of H(2)/2 of 10(9)-10(10) Oe2 cm-1), generated by a ferromagnetic wedge in a uniform magnetic field and rotated on a 1 rpm clinostat. After 4 h 90% of coleoptiles had curved toward the HGMF. The cells affected by HGMF showed clear intracellular displacement of amyloplasts. Coleoptiles in a magnetic field next to a non-ferromagnetic wedge showed no preferential curvature. The small size of the area of nonuniformity of the HGMF allowed mapping of the sensitivity of the coleoptiles by varying the initial position of the wedge relative to the coleoptile apex. When the ferromagnetic wedge was placed 1 mm below the coleoptile tip only 58% of the coleoptiles curved toward the wedge indicating that the cells most sensitive to intracellular displacement of amyloplasts and thus gravity sensing are confined to the top 1 mm portion of barley coleoptiles. Similar experiments with tomato hypocotyls (Lycopersicum esculentum) also resulted in curvature toward the HGMF. The data strongly support the amyloplast-based gravity-sensing system in higher plants and the usefulness of HGMF to substitute gravity in shoots.

Time-lapse analysis of gravitropism in Ceratodon protonemata

NASA Technical Reports Server (NTRS)

Young, J. C.; Sack, F. D.

1992-01-01

The tip cell of the protonema of the moss Ceratodon purpureus (Hedw.) Brid. is negatively gravitropic when grown in the dark on supplemented agar. Gravitropism, plastid distribution, and plastid movement were studied in living cells using time-lapse video microscopy and infrared light. A wrong-way (downward) curvature preceded upward curvature and was detected as early as 2 minutes after reorientation. Upward curvature began 30-45 minutes after reorientation to the horizontal. Cell division temporarily reversed upward curvature, but did not inhibit wrong-way curvature. Since significant amyloplast sedimentation always occurred before the start of upward curvature, it is possible that these amyloplasts function as statoliths for upward curvature. However, no significant amyloplast sedimentation occurred before wrong-way curvature. Thus, this early phase of gravitropism cannot require plastid sedimentation for gravity sensing. Most plastids moved within and between zones, and plastid zonation was highly dynamic. Plastids moved toward the apex and toward the base of the cell at rates much slower than cytoplasmic streaming. Despite the dynamic nature of plastid movement and zonation, during upward curvature the distance between sedimented plastids and the apex stayed constant. Time-lapse analysis has revealed intriguing events not readily seen previously using destructive sampling.

Effects of clinorotation and microgravity on sweet clover columella cells treated with cytochalasin D

NASA Technical Reports Server (NTRS)

Hilaire, E.; Paulsen, A. Q.; Brown, C. S.; Guikema, J. A.; Spooner, B. S. (Principal Investigator)

1995-01-01

The cytoskeleton of columella cells is believed to be involved in maintaining the developmental polarity of cells observed as a reproducible positioning of cellular organelles. It is also implicated in the transduction of gravitropic signals. Roots of sweet clover (Melilotus alba L.) seedlings were treated with a microfilament disrupter, cytochalasin D, on a slowly rotating horizontal clinostat (2 rpm). Electron micrographs of treated columella cells revealed several ultrastructural effects including repositioning of the nucleus and the amyloplasts and the formation of endoplasmic reticulum (ER) whorls. However, experiments performed during fast clinorotation (55 rpm) showed an accumulation (but no whorling) of a disorganized ER network at the proximal and distal pole and a random distribution of the amyloplasts. Therefore, formation of whorls depends upon the speed of clinorotation, and the overall impact of cytochalasin D suggests the necessity of microfilaments in organelle positioning. Interestingly, a similar drug treatment performed in microgravity aboard the US Space Shuttle Endeavour (STS-54, January 1993) caused a displacement of ER membranes and amyloplasts away from the distal plasma membrane. In the present study, we discuss the role of microfilaments in maintaining columella cell polarity and the utility of clinostats to simulate microgravity.

Gravitropism in caulonemata of the moss Pottia intermedia

NASA Technical Reports Server (NTRS)

Chaban, C. I.; Kern, V. D.; Ripetskyj, R. T.; Demkiv, O. T.; Sack, F. D.

1998-01-01

The gravitropism of caulonemata of Pottia intermedia is described and compared with that of other mosses. Spore germination produces primary protonemata including caulonemata which give rise to buds that form the leafy moss plant, the gametophore. Primary caulonemata are negatively gravitropic but their growth and the number of filaments are limited in the dark. Axenic culture of gametophores results in the production of secondary caulonemata that usually arise near the leaf base. Secondary protonemata that form in the light are agravitropic. Secondary caulonemata that form when gametophores are placed in the dark for several days show strong negative gravitropism and grow well in the dark. When upright caulonemata are reorientated to the horizontal or are inverted, upward bending can be detected after 1 h and caulonemata reach the vertical within 1-2 d. Clear amyloplast sedimentation occurs 10-15 minutes after horizontal placement and before the start of upward curvature. This sedimentation takes place in a sub-apical zone. Amyloplast sedimentation also takes place along the length of upright and inverted Pottia protonemata. These results support the hypothesis that amyloplast sedimentation functions in gravitropic sensing since sedimentation occurs before gravitropism in Pottia and since the location and presence of a unique sedimentation zone is conserved in all four mosses known to gravitropic protonomata.

Cyclotron-based effects on plant gravitropism

NASA Astrophysics Data System (ADS)

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.

Maize opaque5 Encodes Monogalactosyldiacylglycerol Synthase and Specifically Affects Galactolipids Necessary for Amyloplast and Chloroplast Function[C][W][OA

PubMed Central

Myers, Alan M.; James, Martha G.; Lin, Qiaohui; Yi, Gibum; Stinard, Philip S.; Hennen-Bierwagen, Tracie A.; Becraft, Philip W.

2011-01-01

The maize (Zea mays) opaque5 (o5) locus was shown to encode the monogalactosyldiacylglycerol synthase MGD1. Null and point mutations of o5 that affect the vitreous nature of mature endosperm engendered an allelic series of lines with stepwise reductions in gene function. C18:3/C18:2 galactolipid abundance in seedling leaves was reduced proportionally, without significant effects on total galactolipid content. This alteration in polar lipid composition disrupted the organization of thylakoid membranes into granal stacks. Total galactolipid abundance in endosperm was strongly reduced in o5- mutants, causing developmental defects and changes in starch production such that the normal simple granules were replaced with compound granules separated by amyloplast membrane. Complete loss of MGD1 function in a null mutant caused kernel lethality owing to failure in both endosperm and embryo development. The data demonstrate that low-abundance galactolipids with five double bonds serve functions in plastid membranes that are not replaced by the predominant species with six double bonds. Furthermore, the data identify a function of amyloplast membranes in the development of starch granules. Finally, the specific changes in lipid composition suggest that MGD1 can distinguish the constituency of acyl groups on its diacylglycerol substrate based upon the degree of desaturation. PMID:21685260

Growth of pea epicotyl in low magnetic field: implication for space research.

PubMed

Negishi, Y; Hashimoto, A; Tsushima, M; Dobrota, C; Yamashita, M; Nakamura, T

1999-01-01

A magnetic field is an inescapable environmental factor for plants on the earth. However, its impact on plant growth is not well understood. In order to survey how magnetic fields affect plant, Alaska pea seedlings were incubated under low magnetic field (LMF) and also in the normal geo-magnetic environment. Two-day-old etiolated seedlings were incubated in a magnetic shield box and in a control box. Sedimentation of amyloplasts was examined in the epicotyls of seedlings grown under these two conditions. The elongation of epicotyls was promoted by LMF. Elongation was most prominent in the middle part of the epicotyls. Cell elongation and increased osmotic pressure of cell sap were found in the epidermal cells exposed to LMF. When the gravitational environment was 1G, the epicotyls incubated under both LMF and normal geomagnetic field grew straight upward and amyloplasts sedimented similarly. However, under simulated microgravity (clinostat), epicotyl and cell elongation was promoted. Furthermore, the epicotyls bent and amyloplasts were dispersed in the cells in simulated microgravity. The dispersion of amyloplasts may relate to the posture control in epicotyl growth under simulated microgravity generated by 3D clinorotation, since it was not observed under LMF in 1G. Since enhanced elongation of cells was commonly seen both at LMF and in simulated microgravity, all elongation on the 3D-clinostat could result from pseudo-low magnetic field, as a by-product of clinorotation. (i.e., clinostat results could be based on randomization of magnetic field together with randomization of gravity vector.) Our results point to the possible use of space for studies in magnetic biology. With space experiments, the effects of dominant environmental factors, such as gravity on plants, could be neutralized or controlled for to reveal magnetic effects more clearly. c1999 COSPAR. Published by Elsevier Science Ltd.

Joining forces: the interface of gravitropism and plastid protein import.

PubMed

Stanga, John; Baldwin, Katherine; Masson, Patrick H

2009-10-01

In flowering plants, gravity perception appears to involve the sedimentation of starch-filled plastids, called amyloplasts, within specialized cells (the statocytes) of shoots (endodermal cells) and roots (columella cells). Unfortunately, how the physical information derived from amyloplast sedimentation is converted into a biochemical signal that promotes organ gravitropic curvature remains largely unknown. Recent results suggest an involvement of the Translocon of the Outer Envelope of (Chloro)plastids (TOC) in early phases of gravity signal transduction within the statocytes. This review summarizes our current knowledge of the molecular mechanisms that govern gravity signal transduction in flowering plants and summarizes models that attempt to explain the contribution of TOC proteins in this important behavioral plant growth response to its mechanical environment.

Altered gravitropic response, amyloplast sedimentation and circumnutation in the Arabidopsis shoot gravitropism 5 mutant are associated with reduced starch levels.

PubMed

Tanimoto, Mimi; Tremblay, Reynald; Colasanti, Joseph

2008-05-01

Plants have developed sophisticated gravity sensing mechanisms to interpret environmental signals that are vital for optimum plant growth. Loss of SHOOT GRAVITROPISM 5 (SGR5) gene function has been shown to affect the gravitropic response of Arabidopsis inflorescence stems. SGR5 is a member of the INDETERMINATE DOMAIN (IDD) zinc finger protein family of putative transcription factors. As part of an ongoing functional analysis of Arabidopsis IDD genes (AtIDD) we have extended the characterisation of SGR5, and show that gravity sensing amyloplasts in the shoot endodermis of sgr5 mutants sediment more slowly than wild type, suggesting a defect in gravity perception. This is correlated with lower amyloplast starch levels, which may account for the reduced gravitropic sensitivity in sgr5. Further, we find that sgr5 mutants have a severely attenuated stem circumnutation movement typified by a reduced amplitude and an decreased periodicity. adg1-1 and sex1-1 mutants, which contain no starch or increased starch, respectively, also show alterations in the amplitude and period of circumnutation. Together these results suggest that plant growth movement may depend on starch levels and/or gravity sensing. Overall, we propose that loss of SGR5 regulatory activity affects starch accumulation in Arabidopsis shoot tissues and causes decreased sensitivity to gravity and diminished circumnutational movements.

Science Notes.

ERIC Educational Resources Information Center

Thurman, Shirley; And Others

1988-01-01

Describes 36 science activities. Topics include: osmosis, fermentation, anhydrobiotic organisms, breathing monitors, trypsin, weeds, amyloplasts, electrolysis, polarimeters, ethene ripening of fruit, colorimetry, diffusion, redox reactions, equilibria, acid-base relationships, electricity, power, resonance, measurement, parallax, amplifiers,…

Characterizing the physics of plant root gravitropism: A systems modeling approach

NASA Astrophysics Data System (ADS)

Yoder, Thomas Lynn

Root gravitropism is divided into three mechanisms; the gravity sensor, transduction, and differential growth. The gravitropic response has been imitated with various mathematical constructs, but a coherent model based on systems engineering concepts does not exist. The goal of this research is to create models of the gravitropic sensor and differential growth response that are consistent with actual physical characteristics of these mechanisms. The study initially establishes that the amyloplasts within the central columella cells of maize are feasible gravity sensors; statoliths. Video-microscopy studies of live root cap sections are used to quantify the dynamics of the statoliths. Extensive MATLAB analysis of amyloplast sedimentation indicates that an actin network interferes with the free sedimentation of the statoliths. This interference is most significant in the central region of the cell and less significant near the periphery. This obstruction of actin creates a channeling behavior in amyloplasts sedimenting through the cell's central region. The amyloplasts also appear to exhibit cross-correlated motions. Cytochalasin D mediates both the channeling and correlated behaviors, confirming that the obstructive influence is actin-based. The video analysis produced a refined value for maize cytoplasmic viscosity. Efforts to model the differential growth mechanism examined historical growth data from numerous researchers. RELEL (relative elemental elongation) growth data applied to a model set analogous to bi-metallic bending is used. Testing and analysis of the model highlights an extremely high sensitivity of curvature to all RELEL parameters. This sensitivity appears to be the reason for the significant differences between gravitropic responses within like species. Newly observed gravitropic responses, along with historical data, are used to explore the gravitropic time response specifications as opposed to averaging individual time-curvature data into single responses. This approach highlights the significant disadvantages of time-averaging, low sampling rates, and a lack of frequency components being incorporated into the response. A single feedback "black box" model is created so that, along with the sensor and differential growth models, some inferences could be made about the elusive transduction mechanism. Numerous pieces of circumstantial evidence are found that indicate that the gravitropic mechanism is not a single-pathway system.

Conservation of the plastid sedimentation zone in all moss genera with known gravitropic protonemata

NASA Technical Reports Server (NTRS)

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

2002-01-01

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.

Plastids and gravitropic sensing

NASA Technical Reports Server (NTRS)

Sack, F. D.

1997-01-01

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.

The structure and development of the starch sheath in pea epicotyls

NASA Technical Reports Server (NTRS)

Sack, D. F.

1985-01-01

Graviperception in plant stems is thought to occur in endodermal cells differentiated as a starch sheath, but little is known about the ultrastructure of these cells in dicots. The structure of the pea starch sheath was studied with respect to gravity and to development in order to determine whether symplastic or apoplastic blockages exist and to describe any intracellular polarity. Amyloplasts increase in size towards the base of the epicotyl hook but are not consistently sedimented until the cells enter the zone exhibiting gravicurvature below the hook. The starch sheath cells are connected to each other and to cells of the cortex and the stele by plasmodesmata. A casparian strip exists in older endodermal cells but not at the stage that the endodermis is differentiated as a starch sheath. Amyloplasts were frequently observed in apparent contact with endoplasmic reticulum.

Amyloplast-Localized SUBSTANDARD STARCH GRAIN4 Protein Influences the Size of Starch Grains in Rice Endosperm1[W

PubMed Central

Matsushima, Ryo; Maekawa, Masahiko; Kusano, Miyako; Kondo, Hideki; Fujita, Naoko; Kawagoe, Yasushi; Sakamoto, Wataru

2014-01-01

Starch is a biologically and commercially important polymer of glucose and is synthesized to form starch grains (SGs) inside amyloplasts. Cereal endosperm accumulates starch to levels that are more than 90% of the total weight, and most of the intracellular space is occupied by SGs. The size of SGs differs depending on the plant species and is one of the most important factors for industrial applications of starch. However, the molecular machinery that regulates the size of SGs is unknown. In this study, we report a novel rice (Oryza sativa) mutant called substandard starch grain4 (ssg4) that develops enlarged SGs in the endosperm. Enlargement of SGs in ssg4 was also observed in other starch-accumulating tissues such as pollen grains, root caps, and young pericarps. The SSG4 gene was identified by map-based cloning. SSG4 encodes a protein that contains 2,135 amino acid residues and an amino-terminal amyloplast-targeted sequence. SSG4 contains a domain of unknown function490 that is conserved from bacteria to higher plants. Domain of unknown function490-containing proteins with lengths greater than 2,000 amino acid residues are predominant in photosynthetic organisms such as cyanobacteria and higher plants but are minor in proteobacteria. The results of this study suggest that SSG4 is a novel protein that influences the size of SGs. SSG4 will be a useful molecular tool for future starch breeding and biotechnology. PMID:24335509

Plastids and carotenoid accumulation

USDA-ARS?s Scientific Manuscript database

Plastids are ubiquitously 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 synth...

Tropism in azalea and lily flowers

NASA Astrophysics Data System (ADS)

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

Tropic responses were examined in azalea Rhododendrom pulchrum and lily Lilium cv. 'Casablanca' flowers. Orientation of the flowers in these two species depicts several up/down characteristics, such as angle of the corolla opening, alignment or configuration of a specific petal at the top, plus direction in the curved tip of the pistil and stamen. Gravity was found to be the prime factor, with light as a secondary signal that determines gravitropism in the pistil of the azalea. Within the azalea, sedimented amyloplasts were observed throughout the cells along the inner layers below the epidermis. In lily flowers, no sedimented amyloplasts were found in style cells, and phototropic responses caused upward bending of the pistil. Responses of lily pistils to monochromatic light were consistent with the action spectrum for phototropism in the shoots of monocotyledonous plants. We discuss how these features may increase the fitness for pollination in these two species.

The response of lazy-2 tomato seedlings to curvature-inducing magnetic gradients is modulated by light

NASA Technical Reports Server (NTRS)

Hasenstein, K. H.; Kuznetsov, O. A.

1999-01-01

Shoots of the lazy-2 mutant of tomato (Lycopersicon esculentum Mill., cv. Ailsa Craig) exhibit negative gravitropism in the dark, but respond positively gravitropically in (red) light. In order to test whether high-gradient magnetic fields (HGMFs) exert only ponderomotive effects on amyloplasts or affect other physiological processes, we induced magnetophoretic curvature in wild-type (WT) and lazy-2 mutant seedlings. Straight hypocotyls of 4-d-old plants were selected and the tips of their hooks were placed in an HGMF near the edge of a magnetized ferromagnetic wedge [grad (H2/2) approximately 10(9)-10(10) Oe2/cm] and mounted on a 1-rpm clinostat. After 4 h in the dark, 85% of WT hypocotyls and 67% of mutant hypocotyls curved toward the wedge. When the seedlings were exposed to red light for 1 h prior to and during the application of the HGMF, 78% of the WT seedlings curved toward the magnetic gradient, but the majority of the lazy-2 seedlings (75%) curved away from the stronger field area. Intracellular amyloplast displacement in the HGMF was similar for both varieties and resembled the displacement after horizontal reorientation. The WT showed a distinct graviresponse pattern depending on the orientation of the hook, even after excision of the apex. Application of HGMFs to decapitated hypocotyls resulted in curvature consistent with that obtained after horizontal reorientation. After light exposure, decapitated lazy-2 seedlings did not respond positively gravitropically. The data imply that the lazy-2 mutants perceive the displacement of amyloplasts in a similar manner to the WT and that the HGMF does not affect the graviresponse mechanism. The study demonstrates that ponderomotive forces due to HGMFs are useful for the analysis of the gravity-sensing mechanism in plants.

Surface Localization of Zein Storage Proteins in Starch Granules from Maize Endosperm1

PubMed Central

Mu-Forster, Chen; Wasserman, Bruce P.

1998-01-01

Starch granules from maize (Zea mays) contain a characteristic group of polypeptides that are tightly associated with the starch matrix (C. Mu-Forster, R. Huang, J.R. Powers, R.W. Harriman, M. Knight, G.W. Singletary, P.L. Keeling, B.P. Wasserman [1996] Plant Physiol 111: 821–829). Zeins comprise about 50% of the granule-associated proteins, and in this study their spatial distribution within the starch granule was determined. Proteolysis of starch granules at subgelatinization temperatures using the thermophilic protease thermolysin led to selective removal of the zeins, whereas granule-associated proteins of 32 kD or above, including the waxy protein, starch synthase I, and starch-branching enzyme IIb, remained refractory to proteolysis. Granule-associated proteins from maize are therefore composed of two distinct classes, the surface-localized zeins of 10 to 27 kD and the granule-intrinsic proteins of 32 kD or higher. The origin of surface-localized δ-zein was probed by comparing δ-zein levels of starch granules obtained from homogenized whole endosperm with granules isolated from amyloplasts. Starch granules from amyloplasts contained markedly lower levels of δ-zein relative to granules prepared from whole endosperm, thus indicating that δ-zein adheres to granule surfaces after disruption of the amyloplast envelope. Cross-linking experiments show that the zeins are deposited on the granule surface as aggregates. In contrast, the granule-intrinsic proteins are prone to covalent modification, but do not form intermolecular cross-links. We conclude that individual granule intrinsic proteins exist as monomers and are not deposited in the form of multimeric clusters within the starch matrix. PMID:9536075

Nodal endoplasmic reticulum, a specialized form of endoplasmic reticulum found in gravity-sensing root tip columella cells

NASA Technical Reports Server (NTRS)

Zheng, H. Q.; Staehelin, L. A.

2001-01-01

The endoplasmic reticulum (ER) of columella root cap cells has been postulated to play a role in gravity sensing. We have re-examined the ultrastructure of columella cells in tobacco (Nicotiana tabacum) root tips preserved by high-pressure freezing/freeze-substitution techniques to gain more precise information about the organization of the ER in such cells. The most notable findings are: the identification of a specialized form of ER, termed "nodal ER," which is found exclusively in columella cells; the demonstration that the bulk of the ER is organized in the form of a tubular network that is confined to a peripheral layer under the plasma membrane; and the discovery that this ER-rich peripheral region excludes Golgi stacks, vacuoles, and amyloplasts but not mitochondria. Nodal ER domains consist of an approximately 100-nm-diameter central rod composed of oblong subunits to which usually seven sheets of rough ER are attached along their margins. These domains form patches at the interface between the peripheral ER network and the ER-free central region of the cells, and they occupy defined positions within central and flanking columella cells. Over one-half of the nodal ER domains are located along the outer tangential walls of the flanking cells. Cytochalasin D and latrunculin A cause an increase in size and a decrease in numbers of nodal ER domains. We postulate that the nodal ER membranes locally modulate the gravisensing signals produced by the sedimenting amyloplasts, and that the confinement of all ER membranes to the cell periphery serves to enhance the sedimentability of the amyloplasts in the central region of columella cells.

Chloroplast redox homeostasis is essential for lateral root formation in Arabidopsis.

PubMed

Ferrández, Julia; González, Maricruz; Cejudo, Francisco Javier

2012-09-01

Redox regulation based on dithiol-disulphide interchange is an essential component of the control of chloroplast metabolism. In contrast to heterotrophic organisms, and non-photosynthetic plant tissues, chloroplast redox regulation relies on ferredoxin (Fd) reduced by the photosynthetic electron transport chain, thus being highly dependent on light. The finding of the NADPH-dependent thioredoxin reductase C (NTRC), a chloroplast-localized NTR with a joint thioredoxin domain, showed that NADPH is also used as source of reducing power for chloroplast redox homeostasis. Recently we have found that NTRC is also in plastids of non-photosynthetic tissues. Because these non-green plastids lack photochemical reactions, their redox homeostasis depends exclusively on NADPH produced from sugars and, thus, NTRC may play an essential role maintaining the redox homeostasis in these plastids. The fact that redox regulation occurs in any type of plastids raises the possibility that the functions of chloroplasts and non-green plastids, such as amyloplasts, are integrated to harmonize the growth of the different organs of the plant. To address this question, we generated Arabidopsis plants the redox homeostasis of which is recovered exclusively in chloroplasts, by leaf-specific expression of NTRC in the ntrc mutant, or exclusively in amyloplasts, by root-specific expression of NTRC. The analysis of these plants suggests that chloroplasts exert a pivotal role on plant growth, as expected because chloroplasts constitute the major source of nutrients and energy, derived from photosynthesis, for growth of heterotrophic tissues. However, NTRC deficiency causes impairment of auxin synthesis and lateral root formation. Interestingly, recovery of redox homeostasis of chloroplasts, but not of amyloplasts, was sufficient to restore wild type levels of lateral roots, showing the important signaling function of chloroplasts for the development of heterotrophic organs.

Cellular polarity and interactions in plant graviperception

NASA Technical Reports Server (NTRS)

Sack, Fred D.

1993-01-01

Presented are results of studies on the mechanisms of gravitropic sensing in higher and lower plants. Gravitropic roots of the aquatic angiosperm, Limnobium, were found to have sedimented amyloplasts in their elongation zone but not in their rootcap; nuclei were found to sediment in the elongation zone as well. Another study attempted to understand how plastid sedimentation occurs in vertical Ceratodon cells and how this sedimentation is regulated. To determine whether the cytoskeleton restricts plastid sedimentation, the effects of amiprophos-methyl (APM) and cytochalasin (CD) on plastid position were qualified. Results 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, demonstrating the importance of the cytoskeleton in maintaining organelle position and cell organization against the force of gravity. Physcomitrella and Funaria were also studied. Results suggest that gravitropism may be relatively common in moss protonemata and reinforce the idea that amyloplast mass functions in gravitropic sensing.

Respiratory processes in non-photosynthetic plastids

PubMed Central

Renato, Marta; Boronat, Albert; Azcón-Bieto, Joaquín

2015-01-01

Chlororespiration is a respiratory process located in chloroplast thylakoids which consists in an electron transport chain from NAD(P)H to oxygen. This respiratory chain involves the NAD(P)H dehydrogenase complex, the plastoquinone pool and the plastid terminal oxidase (PTOX), and it probably acts as a safety valve to prevent the over-reduction of the photosynthetic machinery in stress conditions. The existence of a similar respiratory activity in non-photosynthetic plastids has been less studied. Recently, it has been reported that tomato fruit chromoplasts present an oxygen consumption activity linked to ATP synthesis. Etioplasts and amyloplasts contain several electron carriers and some subunits of the ATP synthase, so they could harbor a similar respiratory process. This review provides an update on the study about respiratory processes in chromoplasts, identifying the major gaps that need to be addressed in future research. It also reviews the proteomic data of etioplasts and amyloplasts, which suggest the presence of a respiratory electron transport chain in these plastids. PMID:26236317

Starch Biosynthesis in Developing Wheat Grain 1

PubMed Central

Keeling, Peter L.; Wood, John R.; Tyson, R. Huw; Bridges, Ian G.

1988-01-01

We have used 13C-labeled sugars and nuclear magnetic resonance (NMR) spectrometry to study the metabolic pathway of starch biosynthesis in developing wheat grain (Triticum aestivum cv Mardler). Our aim was to examine the extent of redistribution of 13C between carbons atoms 1 and 6 of [1-13C] or [6-13C]glucose (or fructose) incorporated into starch, and hence provide evidence for or against the involvement of triose phosphates in the metabolic pathway. Starch synthesis in the endosperm tissue was studied in two experimental systems. First, the 13C sugars were supplied to isolated endosperm tissue incubated in vitro, and second the 13C sugars were supplied in vivo to the intact plant. The 13C starch produced by the endosperm tissue of the grain was isolated and enzymically degraded to glucose using amyloglucosidase, and the distribution of 13C in all glucosyl carbons was quantified by 13C-NMR spectrometry. In all of the experiments, irrespective of the incubation time or incubation conditions, there was a similar pattern of partial (between 15 and 20%) redistribution of label between carbons 1 and 6 of glucose recovered from starch. There was no detectable increase over background 13C incidence in carbons 2 to 5. Within each experiment, the same pattern of partial redistribution of label was found in the glucosyl and fructosyl moieties of sucrose extracted from the tissue. Since it is unlikely that sucrose is present in the amyloplast, we suggest that the observed redistribution of label occurred in the cytosolic compartment of the endosperm cells and that both sucrose and starch are synthesized from a common pool of intermediates, such as hexose phosphate. We suggest that redistribution of label occurs via a cytosolic pathway cycle involving conversion of hexose phosphate to triose phosphate, interconversion of triose phosphate by triose phosphate isomerase, and resynthesis of hexose phosphate in the cytosol. A further round of triose phosphate interconversion in the amyloplast could not be detected. These data seriously weaken the argument for the selective uptake of triose phosphates by the amyloplast as part of the pathway of starch biosynthesis from sucrose in plant storage tissues. Instead, we suggest that a hexose phosphate such as glucose 1-phosphate, glucose 6-phosphate, or fructose 6-phosphate is the most likely candidate for entry into the amyloplast. A pathway of starch biosynthesis is presented, which is consistent with our data and with the current information on the intracellular distribution of enzymes in plant storage tissues. PMID:16666140

How Roots Perceive and Respond to Gravity.

ERIC Educational Resources Information Center

Moore, Randy

1984-01-01

Discusses graviperception and gravitropism by plant roots. Indicates that graviperception occurs via sedimentation of amyloplasts in columella cells of the root cap and that the minimal graviresponsiveness of lateral roots may be due to the intensity of their caps to establish a concentration gradient of inhibitor(s) sufficient to affect…

Involvement of the Vacuoles of the Endodermis in the Early Process of Shoot Gravitropism in Arabidopsis

PubMed Central

Morita, Miyo Terao; Kato, Takehide; Nagafusa, Kiyoshi; Saito, Chieko; Ueda, Takashi; Nakano, Akihiko; Tasaka, Masao

2002-01-01

The endodermal cells of the shoot are thought to be the gravity-sensing cells in Arabidopsis. The amyloplasts in the endodermis that sediment in the direction of gravity may act as statoliths. Endodermis-specific expression of SGR2 and ZIG using the SCR promoter could complement the abnormal shoot gravitropism of the sgr2 and zig mutants, respectively. The abnormalities in amyloplast sedimentation observed in both mutants recovered simultaneously. These results indicate that both genes in the endodermal cell layer are crucial for shoot gravitropism. ZIG encodes AtVTI11, which is a SNARE involved in vesicle transport to the vacuole. The fusion protein of SGR2 and green fluorescent protein localized to the vacuole and small organelles. These observations indicate that ZIG and SGR2 are involved in the formation and function of the vacuole, a notion supported by the results of subcellular analysis of the sgr2 and zig mutants with electron microscopy. These results strongly suggest that the vacuole participates in the early events of gravitropism and that SGR2 and ZIG functions are involved. PMID:11826298

Gravity sensing and signal transduction in vascular plant primary roots.

PubMed

Baldwin, Katherine L; Strohm, Allison K; Masson, Patrick H

2013-01-01

During gravitropism, the potential energy of gravity is converted into a biochemical signal. How this transfer occurs remains one of the most exciting mysteries in plant cell biology. New experiments are filling in pieces of the puzzle. In this review, we introduce gravitropism and give an overview of what we know about gravity sensing in roots of vascular plants, with special highlight on recent papers. When plant roots are reoriented sideways, amyloplast resedimentation in the columella cells is a key initial step in gravity sensing. This process somehow leads to cytoplasmic alkalinization of these cells followed by relocalization of auxin efflux carriers (PINs). This changes auxin flow throughout the root, generating a lateral gradient of auxin across the cap that upon transmission to the elongation zone leads to differential cell elongation and gravibending. We will present the evidence for and against the following players having a role in transferring the signal from the amyloplast sedimentation into the auxin signaling cascade: mechanosensitive ion channels, actin, calcium ions, inositol trisphosphate, receptors/ligands, ARG1/ARL2, spermine, and the TOC complex. We also outline auxin transport and signaling during gravitropism.

Tip-growing cells of the moss Ceratodon purpureus Are gravitropic in high-density media

NASA Technical Reports Server (NTRS)

Schwuchow, Jochen Michael; Kern, Volker Dieter; Sack, Fred David

2002-01-01

Gravity sensing in plants and algae is hypothesized to rely upon either the mass of the entire cell or that of sedimenting organelles (statoliths). Protonemata of the moss Ceratodon purpureus show upward gravitropism and contain amyloplasts that sediment. If moss sensing were whole-cell based, then media denser than the cell should prevent gravitropism or reverse its direction. Cells that were inverted or reoriented to the horizontal displayed distinct negative gravitropism in solutions of iodixanol with densities of 1.052 to 1.320 as well as in bovine serum albumin solutions with densities of 1.037 to 1.184 g cm(-3). Studies using tagged molecules of different sizes and calculations of diffusion times suggest that both types of media penetrate through the apical cell wall. Estimates of the density of the apical cell range from 1.004 to 1.085. Because protonemata grow upward when the cells have a density that is lower than the surrounding medium, gravitropic sensing probably utilizes an intracellular mass in moss protonemata. These data provide additional support for the idea that sedimenting amyloplasts function as statoliths in gravitropism.

Effects of inversion on plastid position and gravitropism in Ceratodon protonemata

NASA Technical Reports Server (NTRS)

Schwuchow, J.; Sack, F. D.

1993-01-01

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.

Retrograde signals arise from reciprocal crosstalk within plastids.

PubMed

Enami, Kazuhiko; Tanaka, Kan; Hanaoka, Mitsumasa

2012-01-01

In addition to the cell nucleus, plant cells also possess genomic DNA and gene expression machineries within mitochondria and plastids. In higher plants, retrograde transcriptional regulation of several nuclear genes encoding plastid-located proteins has been observed in response to changes in a wide variety of physiological properties in plastids, including organelle gene expression (OGE) and tetrapyrrole metabolism. This regulation is postulated to be accomplished by plastid-to-nucleus signaling, (1,2) although the overall signal transduction pathway(s) are not well characterized. By applying a specific differentiation system in tobacco Bright Yellow-2 (BY-2) cultured cells, (3,4) we recently reported that the regulatory system of nuclear gene expressions modulated by a plastid signal was also observed during differentiation of plastids into amyloplasts. (5) While retrograde signaling from plastids was previously speculated to consist of several independent pathways, we found inhibition of OGE and perturbation in the cellular content of one tetrapyrrole intermediate, heme, seemed to interact to regulate amyloplast differentiation. Our results thus highlight the possibility that several sources of retrograde signaling in plastids could be integrated in an intraorganellar manner.

Effects of spaceflight (STS-87) on tropisms and plastid positioning in protonemata of the moss Ceratodon purpureus

NASA Astrophysics Data System (ADS)

Kern, V. D.; Sack, F. D.

Apical cells of moss protonemata represent a single-celled system that perceives and reacts to light (positive and negative phototropism) and to gravity (negative gravitropism). Phototropism completely overrides gravitropism when apical cells are laterally irradiated with relatively high red light intensities, but below a defined light intensity threshold gravitropism competes with the phototropic reaction. A 16 day-long exposure to microgravity conditions demonstrated that gravitropism is allowed when protonemata are laterally illuminated with light intensities below 140 nmol m-2s-1. Protonemata that were grown in darkness in microgravity expressed an endogenous tendency to grow in arcs so that the overall culture morphology resembled a clockwise spiral. However this phenomenon only was observed in cultures that had reached a critical age and/or size. Organelle positioning in dark-grown apical cells was significantly altered in microgravity. Gravisensing most likely involves the sedimentation of starch-filled amyloplasts in a well-defined area of the tip cell. Amyloplasts that at 1-g are sedimented were clustered at the apical part of the sedimentation zone in microgravity. Clustering observed in microgravity or during clino-rotation significantly differs from sedimentation-induced plastid aggregations after inversion of tip cells at 1-g.

Gravisensing, apoptosis, and drug recovery in Taxus cell suspensions

NASA Technical Reports Server (NTRS)

Durzan, D. J.

1999-01-01

Haploid and diploid cell suspensions of Taxus spp. were examined for their adaptive plasticity in response to simulated microgravity, unit gravity, and hypergravity. Cell suspensions produced the taxane, paclitaxel, (TAXOL (R)), which is useful for the treatment of various cancers. Amyloplasts contributed to taxane ring biosynthesis and to drug release at the cell wall. Drug-producing cells reacted as gravisensing osmotic tensiometers. In stressed cells, amyloplasts docked and fused in clusters to sites on the plasmalemma before taxane discharge into the culture medium. In simulated microgravity and compared to all other treatments, taxane production was reduced nearly 100-fold. The percent paclitaxel of total taxanes remained 3-to 6-fold greater, and biomass doubled. When p53-independent programmed cell death was induced, taxanes were released into the culture medium as free molecules (soluble and insoluble) or bound to membranes, nuclear fragments, xylan residues, and other particulate materials. Unit gravity and especially hypergravity promoted xylogenesis and significant drug overproduction. A model relating families of >touch = (TCH), taxane early response (TER), nuclear cycling, and apoptosis-regulating genes to gravisensing, cell wall modifications, and to taxane recovery accounted for most but not all of the observations.

Differentiation and Tropisms in Space-Grown Moss

NASA Technical Reports Server (NTRS)

Sack, Fred D.; Kern, Volker

1999-01-01

This grant supported a Space Shuttle experiment on the effects of microgravity on moss cells. Moss provides a rich system for gravitational and spaceflight research. The early phase of the moss life cycle consists of chains of cells that only grow only at their tips. In the moss Ceratodon purpureus these filaments (protonemata) grow away from gravity in the dark, in a process called gravitropism. The tipmost cells, the apical cells, contain heavy starch-filled bodies called amyloplasts that probably function in g-sensing and that sediment within the apical cell. The SPM-A (Space Moss aka SPAM) experiment flew in November - December, 1997 on STS-87 as part of the Collaborative US Ukrainian Experiment (CLTE). The experiment was accommodated in hardware purpose-built by NASA KSC and Bionetics and included Petri Dish Fixation Units (PDFU) and BRIC-LEDs. Together, this hardware allowed for the culture of the moss on agar in commercial petri dishes, for unilateral illumination with red light of varying intensity, and for chemical fixation in situ. The key findings of the spaceflight were quite unexpected. Neither the orientation of tip-growth nor the distribution of amyloplasts was random in microgravity.

Gravitropism in cut flower stalks of snapdragon

NASA Astrophysics Data System (ADS)

Philosoph-Hadas, S.; Friedman, H.; Meir, S.; Berkovitz-SimanTov, R.; Rosenberger, I.; Halevy, A. H.; Kaufman, P. B.; Balk, P.; Woltering, E. J.

The negative gravitropic response of cut flower stalks is a complex multistep process that requires the participation of various cellular components acting in succession or in parallel. The process was particularly characterized in snapdragon (Antirrhinum majus L.) spikes with regard to (1) gravity stimulus perception associated with amyloplast reorientation; (2) stimulus transduction mediated through differential changes in the level, action and related genes of auxin and ethylene and their possible interaction; (3) stimulus response associated with differential growth leading to stalk curvature; (4) involvement of cytosolic calcium and actin cytoskeleton. Results show that the gravity-induced amyloplast reorientation, differential over-expression of two early auxin responsive genes and asymmetrical distribution of free IAA are early events in the bending process. These precede the asymmetrical ethylene production and differential stem growth, which was derived from initial shrinkage of the upper stem side and a subsequent elongation of the lower stem side. Results obtained with various calcium- and cytoskeleton-related agents indicate that cytosolic calcium and actin filaments may play essential roles in gravitropism-related processes of cut flower stalks. Therefore, modulators of these two physiological mediators may serve as means for controlling any undesired gravitropic bending.

Mapping the functional roles of cap cells in the response of Arabidopsis primary roots to gravity

NASA Technical Reports Server (NTRS)

Blancaflor, E. B.; Fasano, J. M.; Gilroy, S.; Evans, M. L. (Principal Investigator)

1998-01-01

The cap is widely accepted to be the site of gravity sensing in roots because removal of the cap abolishes root curvature. Circumstantial evidence favors the columella cells as the gravisensory cells because amyloplasts (and often other cellular components) are polarized with respect to the gravity vector. However, there has been no functional confirmation of their role. To address this problem, we used laser ablation to remove defined cells in the cap of Arabidopsis primary roots and quantified the response of the roots to gravity using three parameters: time course of curvature, presentation time, and deviation from vertical growth. Ablation of the peripheral cap cells and tip cells did not alter root curvature. Ablation of the innermost columella cells caused the strongest inhibitory effect on root curvature without affecting growth rates. Many of these roots deviated significantly from vertical growth and had a presentation time 6-fold longer than the controls. Among the two inner columella stories, the central cells of story 2 contributed the most to root gravitropism. These cells also exhibited the largest amyloplast sedimentation velocities. Therefore, these results are consistent with the starch-statolith sedimentation hypothesis for gravity sensing.

Pollen tube reuses intracellular components of nucellar cells undergoing programmed cell death in Pinus densiflora.

PubMed

Hiratsuka, Rie; Terasaka, Osamu

2011-04-01

Through the process known as programmed cell death (PCD), nucelli of Pinus densiflora serve as the transmitting tissue for growth of the pollen tube. We sought to clarify the processes of degradation of nucellar cell components and their transport to the pollen tube during PCD in response to pollen tube penetration of such nucelli. Stimulated by pollination, synthesis of large amounts of starch grains occurred in cells in a wide region of the nucellus, but as the pollen tube penetrated the nucellus, starch grains were degraded in amyloplasts of nucellar cells. In cells undergoing PCD, electron-dense vacuoles with high membrane contrast appeared, assumed a variety of autophagic structures, expanded, and ultimately collapsed and disappeared. Vesicles and electron-dense amorphous materials were released inside the thickened walls of cells undergoing PCD, and those vesicles and materials reaching the pollen tube after passing through the extracellular matrix were taken into the tube by endocytosis. These results show that in PCD of nucellar cells, intracellular materials are degraded in amyloplasts and vacuoles, and some of the degraded material is supplied to the pollen tube by vesicular transport to support tube growth.

Curvature induced by amyloplast magnetophoresis in protonemata of the moss Ceratodon purpureus

NASA Technical Reports Server (NTRS)

Kuznetsov, O. A.; Schwuchow, J.; Sack, F. D.; Hasenstein, K. H.

1999-01-01

After gravistimulation of Ceratodon purpureus (Hedw.) Brid. protonemata in the dark, amyloplast sedimentation was followed by upward curvature in the wild-type (WT) and downward curvature in the wwr mutant (wrong way response). We used ponderomotive forces induced by high-gradient magnetic fields (HGMF) to simulate the effect of gravity and displace the presumptive statoliths. The field was applied by placing protonemata either between two permanent magnets at the edge of the gap, close to the edge of a magnetized ferromagnetic wedge, or close to a small (<1 mm) permanent magnet. Continuous application of an HGMF in all three configurations resulted in plastid displacement and induced curvature in tip cells of WT and wwr protonemata. WT cells curved toward the HGMF, and wwr cells curved away from the HGMF, comparable to gravitropism. Plastids isolated from protonemal cultures had densities ranging from 1.24 to 1.38 g cm-3. Plastid density was similar for both genotypes, but the mutant contained larger plastids than the WT. The size difference might explain the stronger response of the wwr protonemata to the HGMF. Our data support the plastid-based theory of gravitropic sensing and suggest that HGMF-induced ponderomotive forces can substitute for gravity.

Gravitropism and phototropism in protonemata of the moss Pohlia nutans (HEDW.) lindb.

NASA Astrophysics Data System (ADS)

Demkiv, O. T.; Kordyum, E. L.; Kardash, O. R.; Khorkavtsiv, O. Ya.

1999-01-01

The gravitropism of protonemata of Pohlia nutans is described and compared with that of other mosses. In darkness, protonemata showed negative gravitropism. Under uniform illumination they grew radially over the substrate surface, whereas unilateral illumination induced positive phototropic growth. Gravitropism was coupled with starch synthesis and amyloplast formation. Protonematal gravitropic growth is more variable than the strict negative gravitropism of Ceratodon chloronema.

Light Promotion of Hypocotyl Gravitropism of a Starch-Deficient Tobacco Mutant Correlates with Plastid Enlargement and Sedimentation1

PubMed Central

Vitha, Stanislav; Yang, Ming; Kiss, John Z.; Sack, Fred D.

1998-01-01

Dark-grown hypocotyls of a starch-deficient mutant (NS458) of tobacco (Nicotiana sylvestris) lack amyloplasts and plastid sedimentation, and have severely reduced gravitropism. However, gravitropism improved dramatically when NS458 seedlings were grown in the light. To determine the extent of this improvement and whether mutant hypocotyls contain sedimented amyloplasts, gravitropic sensitivity (induction time and intermittent stimulation) and plastid size and position in the endodermis were measured in seedlings grown for 8 d in the light. Light-grown NS458 hypocotyls were gravitropic but were less sensitive than the wild type (WT). Starch occupied 10% of the volume of NS458 plastids grown in both the light and the dark, whereas WT plastids were essentially filled with starch in both treatments. Light increased plastid size twice as much in the mutant as in the WT. Plastids in light-grown NS458 were sedimented, presumably because of their larger size and greater total starch content. The induction by light of plastid sedimentation in NS458 provides new evidence for the role of plastid mass and sedimentation in stem gravitropic sensing. Because the mutant is not as sensitive as the WT, NS458 plastids may not have sufficient mass to provide full gravitropic sensitivity. PMID:9490754

Molecular mechanisms of root gravity sensing and signal transduction.

PubMed

Strohm, Allison K; Baldwin, Katherine L; Masson, Patrick H

2012-01-01

Plants use gravity as a guide to direct their roots down into the soil to anchor themselves and to find resources needed for growth and development. In higher plants, the columella cells of the root tip form the primary site of gravity sensing, and in these cells the sedimentation of dense, starch-filled plastids (amyloplasts) triggers gravity signal transduction. This generates an auxin gradient across the root cap that is transmitted to the elongation zone where it promotes differential cell elongation, allowing the root to direct itself downward. It is still not well understood how amyloplast sedimentation leads to auxin redistribution. Models have been proposed to explain how mechanosensitive ion channels or ligand-receptor interactions could connect these events. Although their roles are still unclear, possible second messengers in this process include protons, Ca(2+), and inositol 1,4,5-triphosphate. Upon gravistimulation, the auxin efflux facilitators PIN3 and PIN7 relocalize to the lower side of the columella cells and mediate auxin redistribution. However, evidence for an auxin-independent secondary mechanism of gravity sensing and signal transduction suggests that this physiological process is quite complex. Furthermore, plants must integrate a variety of environmental cues, resulting in multifaceted relationships between gravitropism and other directional growth responses such as hydro-, photo-, and thigmotropism. Copyright © 2011 Wiley Periodicals, Inc.

Curvature Induced by Amyloplast Magnetophoresis in Protonemata of the Moss Ceratodon purpureus1

PubMed Central

Kuznetsov, Oleg A.; Schwuchow, Jochen; Sack, Fred D.; Hasenstein, Karl H.

1999-01-01

After gravistimulation of Ceratodon purpureus (Hedw.) Brid. protonemata in the dark, amyloplast sedimentation was followed by upward curvature in the wild-type (WT) and downward curvature in the wwr mutant (wrong way response). We used ponderomotive forces induced by high-gradient magnetic fields (HGMF) to simulate the effect of gravity and displace the presumptive statoliths. The field was applied by placing protonemata either between two permanent magnets at the edge of the gap, close to the edge of a magnetized ferromagnetic wedge, or close to a small (<1 mm) permanent magnet. Continuous application of an HGMF in all three configurations resulted in plastid displacement and induced curvature in tip cells of WT and wwr protonemata. WT cells curved toward the HGMF, and wwr cells curved away from the HGMF, comparable to gravitropism. Plastids isolated from protonemal cultures had densities ranging from 1.24 to 1.38 g cm−3. Plastid density was similar for both genotypes, but the mutant contained larger plastids than the WT. The size difference might explain the stronger response of the wwr protonemata to the HGMF. Our data support the plastid-based theory of gravitropic sensing and suggest that HGMF-induced ponderomotive forces can substitute for gravity. PMID:9952461

Light promotion of hypocotyl gravitropism of a starch-deficient tobacco mutant correlates with plastid enlargement and sedimentation

NASA Technical Reports Server (NTRS)

Vitha, S.; Yang, M.; Kiss, J. Z.; Sack, F. D.

1998-01-01

Dark-grown hypocotyls of a starch-deficient mutant (NS458) of tobacco (Nicotiana sylvestris) lack amyloplasts and plastid sedimentation, and have severely reduced gravitropism. However, gravitropism improved dramatically when NS458 seedlings were grown in the light. To determine the extent of this improvement and whether mutant hypocotyls contain sedimented amyloplasts, gravitropic sensitivity (induction time and intermittent stimulation) and plastid size and position in the endodermis were measured in seedlings grown for 8 d in the light. Light-grown NS458 hypocotyls were gravitropic but were less sensitive than the wild type (WT). Starch occupied 10% of the volume of NS458 plastids grown in both the light and the dark, whereas WT plastids were essentially filled with starch in both treatments. Light increased plastid size twice as much in the mutant as in the WT. Plastids in light-grown NS458 were sedimented, presumably because of their larger size and greater total starch content. The induction by light of plastid sedimentation in NS458 provides new evidence for the role of plastid mass and sedimentation in stem gravitropic sensing. Because the mutant is not as sensitive as the WT, NS458 plastids may not have sufficient mass to provide full gravitropic sensitivity.

Effect of dark pretreatment on the kinetics of response of barley pulvini to gravistimulation and hormones

NASA Technical Reports Server (NTRS)

Brock, T. G.; Kaufman, P. B.

1988-01-01

Starch in pulvinus amyloplasts of barley (Hordeum vulgare cv Larker) disappears when 45-day-old, light-grown plants are given 5 days of continuous darkness. The effect of this loss on the pulvinus graviresponse was evaluated by following changes in the kinetics of response during the 5-day dark period. Over 5 days of dark pretreatment, the lag to initial graviresponse and the subsequent half-time to maximum steady state bending rate increased significantly while the maximum bending rate did not change. The change in response to applied indoleacetic acid (100 micromolar) plus gibberellic acid (10 micromolar) without gravistimulation, under identical dark pretreatments, was used as a model system for the response component of gravitropism. Dark pretreatment did not change the lag to initial response following hormone application to vertical pulvini, but both the maximum bending rate and the half-time to the maximum rate were significantly reduced. Also, after dark pretreatment, significant bending responses following hormone application were observed in vertical segments with or without added sucrose, while gravistimulation produced a response only if segments were given sucrose. These results indicate that starch-filled amyloplasts are required for the graviresponse of barley pulvini and suggest that they function in the stimulus perception and signal transduction components of gravitropism.

Microgravity and clinorotation cause redistribution of free calcium in sweet clover columella cells

NASA Technical Reports Server (NTRS)

Hilaire, E.; Paulsen, A. Q.; Brown, C. S.; Guikema, J. A.; Spooner, B. S. (Principal Investigator)

1995-01-01

In higher plants, calcium redistribution is believed to be crucial for the root to respond to a change in the direction of the gravity vector. To test the effects of clinorotation and microgravity on calcium localization in higher plant roots, sweet clover (Melilotus alba L.) seedlings were germinated and grown for two days on a slow rotating clinostat or in microgravity on the US Space Shuttle flight STS-60. Subsequently, the tissue was treated with a fixative containing antimonate (a calcium precipitating agent) during clinorotation or in microgravity and processed for electron microscopy. In root columella cells of clinorotated plants, antimonate precipitates were localized adjacent to the cell wall in a unilateral manner. Columella cells exposed to microgravity were characterized by precipitates mostly located adjacent to the proximal and lateral cell wall. In all treatments some punctate precipitates were associated with vacuoles, amyloplasts, mitochondria, and euchromatin of the nucleus. A quantitative study revealed a decreased number of precipitates associated with the nucleus and the amyloplasts in columella cells exposed to microgravity as compared to ground controls. These data suggest that roots perceive a change in the gravitational field, as produced by clinorotation or space flights, and respond respectively differently by a redistribution of free calcium.

Automorphosis of higher plants on a 3-D clinostat

NASA Astrophysics Data System (ADS)

Hoson, T.; Kamisaka, S.; Yamashita, M.; Masuda, Y.

On a three-dimensional (3-D) clinostat, various plant organs developed statocytes capable of responding to the gravity vector. The graviresponse of primary roots of garden cress and maize grown on the clinostat was the same as the control roots, whereas that of maize coleoptiles was reduced. When maize seedlings were grown in the presence of 10^-4 M gibberellic acid and kinetin, the graviresponse of both roots and shoots was suppressed. The corresponding suppression of amyloplast development was observed in the clinostatted and the hormone-treated seedlings. Maize roots and shoots showed spontaneous curvatures in different portions on the 3-D clinostat. The hormone treatment did not significantly influence such an automorphic curvature. When the root cap was removed, maize roots did not curve gravitropically. However, the removal suppressed the automorphic curvatures only slightly. On the other hand, the removal of coleoptile tip did not influence its graviresponse, whereas the spontaneous curvature of decapitated coleoptiles on the clinostat was strongly suppressed. Also, cytochalasin B differently affected the gravitropic and the automorphic curvatures of maize roots and shoots. From these results it is concluded that the graviperception and the early processes of signal transmission are unnecessary for automorphoses under simulated microgravity conditions. Moreover, the results support the view that the amyloplasts act as statoliths probably via an interaction with microfilaments.

Early events in geotropism of seedling shoots

NASA Technical Reports Server (NTRS)

Pickard, B. G.

1985-01-01

Developments during the first ten minutes of geotropic stimulation in plant seedling shoots are reviewed. Topics include induction and curvature; early processes; the relationship between auxin, electric field, calcium, and differential growth; gravity reception leading to Went-Cholodny transport; and comparison of root and shoot. Early processes reviewed are sedimentation of amyloplasts, release of ethylene, rise of electrical and auxin asymmetry, redistribution of calcium, asymmetric vascular transport, increase in tendency to deposit callose, and simulation of putative exocytotic voltage transients.

Effects of the ionic liquid 1-hexyl-3-methylimidazolium bromide on root gravitropism in Arabidopsis seedlings.

PubMed

Zhang, Liang; Wang, Tianqi; Zheng, Fengxia; Ma, Lingyu; Li, Jingyuan

2016-03-01

The toxic effects of ionic liquids (ILs) have attracted increasing attention in recent years. However, the knowledge about the toxic effects of ILs on tropism in organisms remains quite limited. In this study, the effects of 1-hexyl-3-methylimidazolium bromide [C6mim]Br on root gravitropism were evaluated using Arabidopsis seedlings. Our results showed that the root growth and gravity response were significantly inhibited with increasing IL concentration. [C6mim]Br treatment affected the amount and distribution pattern of amyloplasts in root cap compared with controls. The auxin distribution marked with DR5rev::VENUS was altered in IL-treated seedlings. The signal intensity and gene expression of auxin efflux carriers PIN2 and PIN3 were obviously decreased by IL stress. Moreover, as consequences in response to gravity stimulus, the asymmetric DR5 signals in control root apex were impaired by IL treatment. The predominant PIN2 signals along the lower flank of root and PIN3 polarization in columella cells were also significantly reduced in seedlings exposed to IL. Our results suggest that the ionic liquid [C6mim]Br affects the amount and distribution of amyloplasts and disturbs the deployment of PIN2 and PIN3, thus impairing auxin flows in response to gravity stimulus and causing deficient root gravitropism in Arabidopsis seedlings. Copyright © 2015 Elsevier Inc. All rights reserved.

OsBT1 encodes an ADP-glucose transporter involved in starch synthesis and compound granule formation in rice endosperm

PubMed Central

Li, Sanfeng; Wei, Xiangjin; Ren, Yulong; Qiu, Jiehua; Jiao, Guiai; Guo, Xiuping; Tang, Shaoqing; Wan, Jianmin; Hu, Peisong

2017-01-01

Starch is the main storage carbohydrate in higher plants. Although several enzymes and regulators for starch biosynthesis have been characterized, a complete regulatory network for starch synthesis in cereal seeds remains elusive. Here, we report the identification and characterization of the rice Brittle1 (OsBT1) gene, which is expressed specifically in the developing endosperm. The osbt1 mutant showed a white-core endosperm and a significantly lower grain weight than the wild-type. The formation and development of compound starch granules in osbt1 was obviously defective: the amyloplast was disintegrated at early developmental stages and the starch granules were disperse and not compound in the endosperm cells in the centre region of osbt1 seeds. The total starch content and amylose content was decreased and the physicochemical properties of starch were altered. Moreover, the degree of polymerization (DP) of amylopectin in osbt1 was remarkably different from that of wild-type. Map-based cloning of OsBT1 indicated that it encodes a putatively ADP-glucose transporter. OsBT1 coded protein localizes in the amyloplast envelope membrane. Furthermore, the expression of starch synthesis related genes was also altered in the osbt1 mutant. These findings indicate that OsBT1 plays an important role in starch synthesis and the formation of compound starch granules. PMID:28054650

A C2H2-type zinc finger protein, SGR5, is involved in early events of gravitropism in Arabidopsis inflorescence stems.

PubMed

Morita, Miyo T; Sakaguchi, Keitaro; Kiyose, Shin-Ichiro; Taira, Kensuke; Kato, Takehide; Nakamura, Moritaka; Tasaka, Masao

2006-08-01

Plants can sense the direction of gravity and change the growth orientation of their organs. To elucidate the molecular mechanisms of gravity perception and the signal transduction of gravitropism, we have characterized a number of shoot gravitropism (sgr) mutants of Arabidopsis. The sgr5-1 mutant shows reduced gravitropism in the inflorescence stem but its root and hypocotyl have normal gravitropism. SGR5 encodes a zinc finger protein with a coiled-coil motif. The SGR5-GFP fusion protein is localized in the nucleus of Arabidopsis protoplasts, suggesting that SGR5 may act as a transcription factor. Analysis of GUS expression under the control of the SGR5 promoter revealed that SGR5 is mainly expressed in the endodermis, the gravity-sensing tissue in inflorescence stems. Furthermore, the observation that endodermis-specific expression of SGR5 using the SCR promoter in the sgr5-1 mutant restores shoot gravitropism indicates that it could function in the gravity-sensing endodermal cell layer. In contrast to other sgr mutants reported previously, almost all amyloplasts in the endodermal cells of the sgr5-1 mutant sedimented in the direction of gravity. Taken together, our results suggest that SGR5 may be involved in an early event in shoot gravitropism such as gravity perception and/or a signaling process subsequent to amyloplast sedimentation as a putative transcription factor in gravity-perceptive cells.

A Role for the TOC Complex in Arabidopsis Root Gravitropism1[W][OA

PubMed Central

Stanga, John P.; Boonsirichai, Kanokporn; Sedbrook, John C.; Otegui, Marisa S.; Masson, Patrick H.

2009-01-01

Arabidopsis (Arabidopsis thaliana) roots perceive gravity and reorient their growth accordingly. Starch-dense amyloplasts within the columella cells of the root cap are important for gravitropism, and starchless mutants such as pgm1 display an attenuated response to gravistimulation. The altered response to gravity1 (arg1) mutant is known to be involved with the early phases of gravity signal transduction. arg1 responds slowly to gravistimulation and is in a genetically distinct pathway from pgm1, as pgm1 mutants enhance the gravitropic defect of arg1. arg1 seeds were mutagenized with ethylmethane sulfonate to identify new mutants that enhance the gravitropic defect of arg1. Two modifier of arg1 mutants (mar1 and mar2) grow in random directions only when arg1 is present, do not affect phototropism, and respond like the wild type to application of phytohormones. Both have mutations affecting different components of the Translocon of Outer Membrane of Chloroplasts (TOC) complex. mar1 possesses a mutation in the TOC75-III gene; mar2 possesses a mutation in the TOC132 gene. Overexpression of TOC132 rescues the random growth phenotype of mar2 arg1 roots. Root cap amyloplasts in mar2 arg1 appear ultrastructurally normal. They saltate like the wild type and sediment at wild-type rates upon gravistimulation. These data point to a role for the plastidic TOC complex in gravity signal transduction within the statocytes. PMID:19211693

A role for the TOC complex in Arabidopsis root gravitropism.

PubMed

Stanga, John P; Boonsirichai, Kanokporn; Sedbrook, John C; Otegui, Marisa S; Masson, Patrick H

2009-04-01

Arabidopsis (Arabidopsis thaliana) roots perceive gravity and reorient their growth accordingly. Starch-dense amyloplasts within the columella cells of the root cap are important for gravitropism, and starchless mutants such as pgm1 display an attenuated response to gravistimulation. The altered response to gravity1 (arg1) mutant is known to be involved with the early phases of gravity signal transduction. arg1 responds slowly to gravistimulation and is in a genetically distinct pathway from pgm1, as pgm1 mutants enhance the gravitropic defect of arg1. arg1 seeds were mutagenized with ethylmethane sulfonate to identify new mutants that enhance the gravitropic defect of arg1. Two modifier of arg1 mutants (mar1 and mar2) grow in random directions only when arg1 is present, do not affect phototropism, and respond like the wild type to application of phytohormones. Both have mutations affecting different components of the Translocon of Outer Membrane of Chloroplasts (TOC) complex. mar1 possesses a mutation in the TOC75-III gene; mar2 possesses a mutation in the TOC132 gene. Overexpression of TOC132 rescues the random growth phenotype of mar2 arg1 roots. Root cap amyloplasts in mar2 arg1 appear ultrastructurally normal. They saltate like the wild type and sediment at wild-type rates upon gravistimulation. These data point to a role for the plastidic TOC complex in gravity signal transduction within the statocytes.

Gravitropism of axial organs in multicellular plants

NASA Astrophysics Data System (ADS)

Kutschera, U.

Gravitropism of plant organs such as roots, stems and coleoptiles can be separated into four distinct phases: 1. perception (gravity sensing), 2. transduction of a signal into the target region and 3. the response (differential growth). This last reaction is followed by a straightening of the curved organ (4.). The perception of the gravitropic stimulus upon horizontal positioning of the organ (1.) occurs via amyloplasts that sediment within the statocytes. This conclusion is supported by our finding that submerged rice coleoptiles that lack sedimentable amyloplasts show no graviresponse. The mode of signal transduction (2.) from the statocytes to the peripheral cell layers is still unknown. Differential growth (3.) consists of a cessation of cell expansion on the upper side and an enhancement of elongation on the lower side of the organ. Based on the facts that the sturdy outer epidermal wall (OEW) constitutes the growth-controlling structure of the coleoptile and that growth-related osmiophilic particles accumulate on the upper OEW, it is concluded that the differential incorporation of wall material (presumably glycoproteins) is causally involved. During gravitropic bending, electron-dense particles ('wall-loosening capacity') accumulate on the growth-inhibited upper OEW. It is proposed that the autotropic straightening response, which is in part due to an acceleration of cell elongation on the curved upper side, may be attributable to an incorporation of the accumulated particles ('release of wall-loosening capacity'). This novel mechanism of autotropic re-bending and its implications for the Cholodny-Went hypothesis are discussed.

Identification of Multiple Phosphorylation Sites on Maize Endosperm Starch Branching Enzyme IIb, a Key Enzyme in Amylopectin Biosynthesis

PubMed Central

Makhmoudova, Amina; Williams, Declan; Brewer, Dyanne; Massey, Sarah; Patterson, Jenelle; Silva, Anjali; Vassall, Kenrick A.; Liu, Fushan; Subedi, Sanjeena; Harauz, George; Siu, K. W. Michael; Tetlow, Ian J.; Emes, Michael J.

2014-01-01

Starch branching enzyme IIb (SBEIIb) plays a crucial role in amylopectin biosynthesis in maize endosperm by defining the structural and functional properties of storage starch and is regulated by protein phosphorylation. Native and recombinant maize SBEIIb were used as substrates for amyloplast protein kinases to identify phosphorylation sites on the protein. A multidisciplinary approach involving bioinformatics, site-directed mutagenesis, and mass spectrometry identified three phosphorylation sites at Ser residues: Ser649, Ser286, and Ser297. Two Ca2+-dependent protein kinase activities were partially purified from amyloplasts, termed K1, responsible for Ser649 and Ser286 phosphorylation, and K2, responsible for Ser649 and Ser297 phosphorylation. The Ser286 and Ser297 phosphorylation sites are conserved in all plant branching enzymes and are located at opposite openings of the 8-stranded parallel β-barrel of the active site, which is involved with substrate binding and catalysis. Molecular dynamics simulation analysis indicates that phospho-Ser297 forms a stable salt bridge with Arg665, part of a conserved Cys-containing domain in plant branching enzymes. Ser649 conservation appears confined to the enzyme in cereals and is not universal, and is presumably associated with functions specific to seed storage. The implications of SBEIIb phosphorylation are considered in terms of the role of the enzyme and the importance of starch biosynthesis for yield and biotechnological application. PMID:24550386

The Arabidopsis LAZY1 Family Plays a Key Role in Gravity Signaling within Statocytes and in Branch Angle Control of Roots and Shoots[OPEN

PubMed Central

Taniguchi, Masatoshi; Furutani, Masahiko; Nishimura, Takeshi; Nakamura, Moritaka; Fushita, Toyohito; Iijima, Kohta; Baba, Kenichiro; Toyota, Masatsugu

2017-01-01

During gravitropism, the directional signal of gravity is perceived by gravity-sensing cells called statocytes, leading to asymmetric distribution of auxin in the responding organs. To identify the genes involved in gravity signaling in statocytes, we performed transcriptome analyses of statocyte-deficient Arabidopsis thaliana mutants and found two candidates from the LAZY1 family, AtLAZY1/LAZY1-LIKE1 (LZY1) and AtDRO3/AtNGR1/LZY2. We showed that LZY1, LZY2, and a paralog AtDRO1/AtNGR2/LZY3 are redundantly involved in gravitropism of the inflorescence stem, hypocotyl, and root. Mutations of LZY genes affected early processes in gravity signal transduction without affecting amyloplast sedimentation. Statocyte-specific expression of LZY genes rescued the mutant phenotype, suggesting that LZY genes mediate gravity signaling in statocytes downstream of amyloplast displacement, leading to the generation of asymmetric auxin distribution in gravity-responding organs. We also found that lzy mutations reversed the growth angle of lateral branches and roots. Moreover, expression of the conserved C-terminal region of LZY proteins also reversed the growth direction of primary roots in the lzy mutant background. In lateral root tips of lzy multiple mutants, asymmetric distribution of PIN3 and auxin response were reversed, suggesting that LZY genes regulate the direction of polar auxin transport in response to gravity through the control of asymmetric PIN3 expression in the root cap columella. PMID:28765510

Proteins from Multiple Metabolic Pathways Associate with Starch Biosynthetic Enzymes in High Molecular Weight Complexes: A Model for Regulation of Carbon Allocation in Maize Amyloplasts1[C][W][OA

PubMed Central

Hennen-Bierwagen, Tracie A.; Lin, Qiaohui; Grimaud, Florent; Planchot, Véronique; Keeling, Peter L.; James, Martha G.; Myers, Alan M.

2009-01-01

Starch biosynthetic enzymes from maize (Zea mays) and wheat (Triticum aestivum) amyloplasts exist in cell extracts in high molecular weight complexes; however, the nature of those assemblies remains to be defined. This study tested the interdependence of the maize enzymes starch synthase IIa (SSIIa), SSIII, starch branching enzyme IIb (SBEIIb), and SBEIIa for assembly into multisubunit complexes. Mutations that eliminated any one of those proteins also prevented the others from assembling into a high molecular mass form of approximately 670 kD, so that SSIII, SSIIa, SBEIIa, and SBEIIb most likely all exist together in the same complex. SSIIa, SBEIIb, and SBEIIa, but not SSIII, were also interdependent for assembly into a complex of approximately 300 kD. SSIII, SSIIa, SBEIIa, and SBEIIb copurified through successive chromatography steps, and SBEIIa, SBEIIb, and SSIIa coimmunoprecipitated with SSIII in a phosphorylation-dependent manner. SBEIIa and SBEIIb also were retained on an affinity column bearing a specific conserved fragment of SSIII located outside of the SS catalytic domain. Additional proteins that copurified with SSIII in multiple biochemical methods included the two known isoforms of pyruvate orthophosphate dikinase (PPDK), large and small subunits of ADP-glucose pyrophosphorylase, and the sucrose synthase isoform SUS-SH1. PPDK and SUS-SH1 required SSIII, SSIIa, SBEIIa, and SBEIIb for assembly into the 670-kD complex. These complexes may function in global regulation of carbon partitioning between metabolic pathways in developing seeds. PMID:19168640

Demonstration of prominent actin filaments in the root columella

NASA Technical Reports Server (NTRS)

Collings, D. A.; Zsuppan, G.; Allen, N. S.; Blancaflor, E. B.; Brown, C. S. (Principal Investigator)

2001-01-01

The distribution of actin filaments within the gravity-sensing columella cells of plant roots remains poorly understood, with studies over numerous years providing inconsistent descriptions of actin organization in these cells. This uncertainty in actin organization, and thus in actin's role in graviperception and gravisignaling, has led us to investigate actin arrangements in the columella cells of Zea mays L., Medicago truncatula Gaertn., Linum usitatissiilium L. and Nicotianla benthamiana Domin. Actin organization was examined using a combination of optimized immunofluorescence techniques, and an improved fluorochrome-conjugated phalloidin labeling method reliant on 3-maleimidobenzoyl-N-hydroxy-succinimide ester (MBS) cross-linking combined with glycerol permeabilization. Confocal microscopy of root sections labeled with anti-actin antibodies revealed patterns suggestive of actin throughout the columella region. These patterns included short and fragmented actin bundles, fluorescent rings around amyloplasts and intense fluorescence originating from the nucleus. Additionally, confocal microscopy of MBS-stabilized and Alexa Fluor-phalloidin-labeled root sections revealed a previously undetected state of actin organization in the columella. Discrete actin structures surrounded the amyloplasts and prominent actin cables radiated from the nuclear surface toward the cell periphery. Furthermore, the cortex of the columella cells contained fine actin bundles (or single filaments) that had a predominant transverse orientation. We also used confocal microscopy of plant roots expressing endoplasmic reticulum (ER)-targeted green fluorescent protein to demonstrate rapid ER movements within the columella cells, suggesting that the imaged actin network is functional. The successful identification of discrete actin structures in the root columella cells forms the perception and signaling.

Transformation of plastids in soil-shaded lowermost hypocotyl segments of bean (Phaseolus vulgaris) during a 60-day cultivation period.

PubMed

Kakuszi, Andrea; Solymosi, Katalin; Böddi, Béla

2017-04-01

The maintenance but substantial transformation of plastids was found in lowermost hypocotyl segments of soil-grown bean plants (Phaseolus vulgaris cv. Magnum) during a 60-day cultivation period. Although the plants were grown under natural light-dark cycles, this hypocotyl segment was under full coverage of the soil in 5-7 cm depth, thus it was never exposed to light. The 4-day-old plants were fully etiolated: amyloplasts, occasionally prolamellar bodies, protochlorophyllide (Pchlide) and protochlorophyll (Pchl) were found in the hypocotyls of these young seedlings. The 633 and 654 nm bands in the 77 K fluorescence emission spectra indicated the presence of Pchlide and Pchl pigments. During aging, both the Pchlide and Pchl contents increased, however, the Pchl to Pchlide ratio gradually increased. In parallel, the contribution of the 654 nm form decreased and in the spectra of the 60-day-old samples, the main band shifted to 631 nm, and a new form appeared with an emission maximum at 641 nm. The photoactivity had been lost; bleaching took place at continuous illumination. The inner membranes of the plastids disappeared, the amount of starch storing amyloplasts decreased. These data may indicate the general importance of plastids for plant cell metabolism, which can be the reason for their maintenance. Also the general heterogeneity of plastid forms can be concluded: in tissues not exposed to light, Pchl accumulating plastids develop and are maintained even for a long period. © 2016 Scandinavian Plant Physiology Society.

The Arabidopsis LAZY1 Family Plays a Key Role in Gravity Signaling within Statocytes and in Branch Angle Control of Roots and Shoots.

PubMed

Taniguchi, Masatoshi; Furutani, Masahiko; Nishimura, Takeshi; Nakamura, Moritaka; Fushita, Toyohito; Iijima, Kohta; Baba, Kenichiro; Tanaka, Hirokazu; Toyota, Masatsugu; Tasaka, Masao; Morita, Miyo Terao

2017-08-01

During gravitropism, the directional signal of gravity is perceived by gravity-sensing cells called statocytes, leading to asymmetric distribution of auxin in the responding organs. To identify the genes involved in gravity signaling in statocytes, we performed transcriptome analyses of statocyte-deficient Arabidopsis thaliana mutants and found two candidates from the LAZY1 family, AtLAZY1 / LAZY1-LIKE1 ( LZY1 ) and AtDRO3 / AtNGR1 / LZY2 We showed that LZY1 , LZY2 , and a paralog AtDRO1/AtNGR2/LZY3 are redundantly involved in gravitropism of the inflorescence stem, hypocotyl, and root. Mutations of LZY genes affected early processes in gravity signal transduction without affecting amyloplast sedimentation. Statocyte-specific expression of LZY genes rescued the mutant phenotype, suggesting that LZY genes mediate gravity signaling in statocytes downstream of amyloplast displacement, leading to the generation of asymmetric auxin distribution in gravity-responding organs. We also found that lzy mutations reversed the growth angle of lateral branches and roots. Moreover, expression of the conserved C-terminal region of LZY proteins also reversed the growth direction of primary roots in the lzy mutant background. In lateral root tips of lzy multiple mutants, asymmetric distribution of PIN3 and auxin response were reversed, suggesting that LZY genes regulate the direction of polar auxin transport in response to gravity through the control of asymmetric PIN3 expression in the root cap columella. © 2017 American Society of Plant Biologists. All rights reserved.

Salt modulates gravity signaling pathway to regulate growth direction of primary roots in Arabidopsis.

PubMed

Sun, Feifei; Zhang, Wensheng; Hu, Haizhou; Li, Bao; Wang, Youning; Zhao, Yankun; Li, Kexue; Liu, Mengyu; Li, Xia

2008-01-01

Plant root architecture is highly plastic during development and can adapt to many environmental stresses. The proper distribution of roots within the soil under various conditions such as salinity, water deficit, and nutrient deficiency greatly affects plant survival. Salinity profoundly affects the root system architecture of Arabidopsis (Arabidopsis thaliana). However, despite the inhibitory effects of salinity on root length and the number of roots, very little is known concerning influence of salinity on root growth direction and the underlying mechanisms. Here we show that salt modulates root growth direction by reducing the gravity response. Exposure to salt stress causes rapid degradation of amyloplasts in root columella cells of Arabidopsis. The altered root growth direction in response to salt was found to be correlated with PIN-FORMED2 (PIN2) messenger RNA abundance and expression and localization of the protein. Furthermore, responsiveness to gravity of salt overly sensitive (sos) mutants is substantially reduced, indicating that salt-induced altered gravitropism of root growth is mediated by ion disequilibrium. Mutation of SOS genes also leads to reduced amyloplast degradation in root tip columella cells and the defects in PIN2 gene expression in response to salt stress. These results indicate that the SOS pathway may mediate the decrease of PIN2 messenger RNA in salinity-induced modification of gravitropic response in Arabidopsis roots. Our findings provide new insights into the development of a root system necessary for plant adaptation to high salinity and implicate an important role of the SOS signaling pathway in this process.

Cytochalasin D does not inhibit gravitropism in roots

NASA Technical Reports Server (NTRS)

Staves, M. P.; Wayne, R.; Leopold, A. C.

1997-01-01

It is generally thought that sedimenting plastids are responsible for gravity sensing in higher plants. We directly tested the model generated by the current statolith hypothesis that the gravity sensing that leads to gravitropism results from an interaction between the plastids and actin microfilaments. We find that the primary roots of rice, corn, and cress undergo normal gravitropism and growth even when exposed to cytochalasin D, a disruptor of actin microfilaments. These results indicate that an interaction between amyloplasts and the actin cytoskeleton is not critical for gravity sensing in higher plants and weaken the current statolith hypothesis.

Plastids and Carotenoid Accumulation.

PubMed

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.

Statolith action by the numbers: Physics and feasbility

NASA Astrophysics Data System (ADS)

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

2016-07-01

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 does the statolith exert on the interface between the ER cortex and inner cytoplasm? What is the origin of the elastic force that opposes the penetration of the statolith and that causes it to rebound when the gravitational force is relieved? Candidates considered included actin filaments, endoplasmic membrane, an immiscible-phase interface, and a miscible-phase interface. Forces estimated to produce a 300nm indentation in such interfaces were 100, 30, 0.3, and <1 pN, respectively. The gravitational force on a single statolith is typically 0.1 pN or less. These numbers suggest that the statolith is interacting with an aqueous-aqueous interface with low interfacial tension (<1 μN/m). Further quantitative testing of these notions should be performed on forthcoming low-gravity sub-orbital crewed flights on which dynamic microscopy can be performed during several seconds of increased g-loading followed by some ten minutes of reduced gravity.

A Comprehensive Analysis of Chromoplast Differentiation Reveals Complex Protein Changes Associated with Plastoglobule Biogenesis and Remodeling of Protein Systems in Sweet Orange Flesh1[OPEN

PubMed Central

Wang, Lun; Deng, Xiuxin

2015-01-01

Globular and crystalloid chromoplasts were observed to be region specifically formed in sweet orange (Citrus sinensis) flesh and converted from amyloplasts during fruit maturation, which was associated with the composition of specific carotenoids and the expression of carotenogenic genes. Subsequent isobaric tag for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analyses of purified plastids from the flesh during chromoplast differentiation and senescence identified 1,386 putative plastid-localized proteins, 1,016 of which were quantified by spectral counting. The iTRAQ values reflecting the expression abundance of three identified proteins were validated by immunoblotting. Based on iTRAQ data, chromoplastogenesis appeared to be associated with three major protein expression patterns: (1) marked decrease in abundance of the proteins participating in the translation machinery through ribosome assembly; (2) increase in abundance of the proteins involved in terpenoid biosynthesis (including carotenoids), stress responses (redox, ascorbate, and glutathione), and development; and (3) maintenance of the proteins for signaling and DNA and RNA. Interestingly, a strong increase in abundance of several plastoglobule-localized proteins coincided with the formation of plastoglobules in the chromoplast. The proteomic data also showed that stable functioning of protein import, suppression of ribosome assembly, and accumulation of chromoplast proteases are correlated with the amyloplast-to-chromoplast transition; thus, these processes may play a collective role in chromoplast biogenesis and differentiation. By contrast, the chromoplast senescence process was inferred to be associated with significant increases in stress response and energy supply. In conclusion, this comprehensive proteomic study identified many potentially new plastid-localized proteins and provides insights into the potential developmental and molecular mechanisms underlying chromoplast biogenesis, differentiation, and senescence in sweet orange flesh. PMID:26056088

Floral nectar production and nectary structure of a bee-pollinated shrub from Neotropical savanna.

PubMed

Guimarães, E; Nogueira, A; Machado, S R

2016-01-01

Biotic pollination is critical for tropical ecosystem functioning, and nectar plays an essential role as it represents the main trophic resource for pollinators. Nevertheless, little is known about the mechanisms that underlie its production, which is essential for understanding the basis of nectar-mediated interactions in ecological and evolutionary approaches. Therefore, this study explores the relationship between the nectar secretion pattern and nectary functional changes in Anemopaegma album, a bee-pollinated species. We analysed the pattern of nectar production under field conditions and investigated floral nectary structural changes in two different developmental stages using light, transmission and scanning electron microscopy. We measured 30.95 ± 23.02 μl (mean ± SD, n = 30) of nectar accumulated inside the nectar chamber (29.26 ± 3.48% sucrose equivalents) at the moment of flower opening. Nectar removal did not influence the pattern of floral nectar production in terms of volume or total sugar but reduced the concentration of the nectar produced during the first 24 h of anthesis. The nectary consisted of an epidermis, a nectary parenchyma and a subnectary parenchyma supplied only by phloem. Starch grains decreased in size and abundance from the subnectary parenchyma toward the epidermis. We observed the degradation of starch grains and incorporation of amyloplasts into vacuoles at the pre-anthesis stage as well as the transformation of amyloplasts into elaioplasts during anthesis. Nectar secretion was continuous during the A. album flower life span, which was related to the functional features of its floral nectary, especially the presence of starch stored in the parenchyma. © 2015 German Botanical Society and The Royal Botanical Society of the Netherlands.

Cytoplasmic pH dynamics in maize pulvinal cells induced by gravity vector changes

NASA Technical Reports Server (NTRS)

Johannes, E.; Collings, D. A.; Rink, J. C.; Allen, N. S.; Brown, C. S. (Principal Investigator)

2001-01-01

In maize (Zea mays) and other grasses, changes in orientation of stems are perceived by pulvinal tissue, which responds to the stimulus by differential growth resulting in upward bending of the stem. The amyloplast-containing bundle sheath cells are the sites of gravity perception, although the initial steps of gravity perception and transmission remain unclear. In columella cells of Arabidopsis roots, we previously found that cytoplasmic pH (pH(c)) is a mediator in early gravitropic signaling (A.C. Scott, N.S. Allen [1999] Plant Physiol 121: 1291-1298). The question arises whether pH(c) has a more general role in signaling gravity vector changes. Using confocal ratiometric imaging and the fluorescent pH indicator carboxy seminaphtorhodafluor acetoxymethyl ester acetate, we measured pH(c) in the cells composing the maize pulvinus. When stem slices were gravistimulated and imaged on a horizontally mounted confocal microscope, pH(c) 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. These changes were most apparent in cells exhibiting net amyloplast sedimentation. Parenchyma cells and isolated bundle sheath cells did not show any gravity-induced pH(c) changes although all cell types responded to external stimuli in the predicted way: Propionic acid and auxin treatments induced acidification, whereas raising the external pH caused alkalinization. The results suggest that pH(c) has an important role in the early signaling pathways of maize stem gravitropism.

Ultrastructure of the root cap of Arabidopsis Thaliana L. Heynh under spaceflight conditions

NASA Technical Reports Server (NTRS)

1983-01-01

Peculiarities of the ultrastructural organization of Arabidopsis root cap cells grown from the stage of two cotyledonous leaves in the Svetoblok-1 apparatus aboard the Salyut 6 research orbital station and in the laboratory are assessed. It is established that under conditions of real space flight vacuolization of the root cap cells increses considerably compared to the control variant. Changes in the topography and ulstrastructure of amyloplasts as well as lysis of cell walls are observed in the material under study. An assumption is advanced on analogous cell responses observed at the ultrastructural level to weightlessness and clinostatic conditions.

Induction of Plant Curvature by Magnetophoresis and Cytoskeletal Changes during Root Graviresponse

NASA Technical Reports Server (NTRS)

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

1996-01-01

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.

Quantitative proteomic analysis of intact plastids.

PubMed

Shiraya, Takeshi; Kaneko, Kentaro; Mitsui, Toshiaki

2014-01-01

Plastids are specialized cell organelles in plant cells that are differentiated into various forms including chloroplasts, chromoplasts, and amyloplasts, and fulfill important functions in maintaining the overall cell metabolism and sensing environmental factors such as sunlight. It is therefore important to grasp the mechanisms of differentiation and functional changes of plastids in order to enhance the understanding of vegetality. In this chapter, details of a method for the extraction of intact plastids that makes analysis possible while maintaining the plastid functions are provided; in addition, a quantitative shotgun method for analyzing the composition and changes in the content of proteins in plastids as a result of environmental impacts is described.

Gravitropism of cut shoots is mediated by oxidative processes: A physiological and molecular study

NASA Astrophysics Data System (ADS)

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

2012-07-01

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 in favor of the lower shoot side was completely eliminated in the NAC-treated shoots, in which the bending response was inhibited. These results suggest that the gravitropic-induced asymmetric auxin redistribution precedes shoot bending, and NAC inhibits this response by abolishing the auxin gradient across the shoot. Our microarray analysis of tomato shoots, using the Affymetrix Tomato GeneChip, revealed differential changes in expression of 266 genes, occurring during the initial 0.5-5 h of shoot reorientation prior to bending. The differential changes in expression of auxin-related genes in favor of the lower shoot side occurred already following 0.5 h of reorientation, while those of cell wall-related genes, associated with shoot bending, occurred only 3 h following shoot reorientation. Among the identified genes, 11 genes were related to auxin, and 36 genes were associated with oxidative processes, demonstrating the involvement of ROS in early events of shoot gravitropism. Taken together, our results suggest that ROS mediate the early gravity-induced amyloplast sedimentation required for signal perception, as well as the lateral auxin movement across the shoot, which is necessary for the auxin asymmetric distribution leading to shoot upward bending.

How roots perceive and respond to gravity

NASA Technical Reports Server (NTRS)

Moore, R.; Evans, M. L.

1986-01-01

Graviperception by plant roots is believed to occur via the sedimentation of amyloplasts in columella cells of the root cap. This physical stimulus results in an accumulation of calcium on the lower side of the cap, which in turn induces gravicurvature. In this paper we present a model for root gravitropism integrating gravity-induced changes in electrical potential, cytochemical localization of calcium in cells of gravistimulated roots, and the interdependence of calcium and auxin movement. Key features of the model are that 1) gravity-induced redistribution of calcium is an early event in the transduction mechanism, and 2) apoplastic movement of calcium through the root-cap mucilage may be an important component of the pathway for calcium movement.

Root cytoskeleton: its role in perception of and response to gravity

NASA Technical Reports Server (NTRS)

Baluska, F.; Hasenstein, K. H.

1997-01-01

We have critically evaluated the possible functions of the plant cytoskeleton in root gravisensing and graviresponse and discussed the evidence that microtubules (MTs) and actin microfilaments (MFs) do not control differential cell growth during bending of roots. On the other hand, MF and MT networks are envisaged to participate in gravisensing because of the mechanical properties of the cytoskeletal structures that interconnect plant cell organelles with the plasma membrane. In restrained gravisensing, forces are suggested to be transmitted to membranes because large-scale gravity-dependent repositioning of organelles is effectively prevented due to the cytoskeleton-mediated anchorage of their envelopes at the plasma membrane. From the cytoskeletal point of view, we can also envisage an unrestrained gravity sensing when cytoskeletal tethers are not strong enough to preserve the tight control over distribution of organelles and the latter, if heavy enough, are allowed to sediment towards the physical bottom of cells. This situation obviously occurs in root cap statocytes because these uniquely organized cells are depleted of prominent actin MF bundles, endoplasmic MT arrays, and ER elements in their internal cytoplasm. Nevertheless, indirect evidence clearly indicates that sedimented root cap statoliths are enmeshed within fine but dynamic MF networks and that their behaviour is obviously under, at least partial, cytoskeletal control. The actomyosin-enriched domain among and around amyloplasts is proposed to increase the perception of gravity due to the grouping effect of sedimenting statoliths. Cytoskeletal links between myosin-rich statoliths, and cell peripheries well equipped with dense cortical MTs, membrane-associated cytoskeleton, as well as with ER elements, would allow efficient restrained gravisensing only at the statocyte cell cortex. As a consequence of cytoskeletal depletion in the internal statocyte cytoplasm and bulk sedimentation of large amyloplasts, restrained gravisensing is spatially restricted to the bottom of the statocyte irrespective of whether roots are vertical or horizontal. This spatial aspect allows for efficient gravisensing via amplification of gravity-induced impacts on the cellular architecture, a phenomenon which is unique to root cap statocytes.

Genetic Analysis of Gravity Signal Transduction in Arabidopsis Roots

NASA Astrophysics Data System (ADS)

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 that the protein-import function of the complex, not the presence of a large acidic domain of TOC132 within the cytoplasm, is needed for gravity signal transduction. Furthermore, mutations in several genes encoding distinct members of the TOC complex also enhanced the gravitropic defect of arg1. Together, these data suggest that the TOC complex works indirectly in gravity signal transduction through its ability to target specific cytoplasmically synthesized proteins, possibly gravity signal transducers, into the plastid. We have used a proteomic strategy to identify root-tip proteins that are differentially expressed between wild type and mar2 mutant plants. The corresponding list of differentially expressed proteins, which includes a surprisingly small number of plastid-targeted molecules, mainly contains proteins that are predicted to be associated with distinct cellular compartments. Several of the corresponding genes were found to also be differentially expressed between wild type and mar2 mutant root tips at the transcriptional level, suggesting cross-talk between amyloplasts and nucleus in these cells. Some of the differentially represented proteins are encoded by genes that are differentially expressed in the root tip in response to gravistimulation, further suggesting their contribution to gravity signal transduction. Work in underway to elucidate their function and potential contribution to this pathway. This work was funded by grants from the National Science Foundation.

Genetic analysis of gravity signal transduction in roots

NASA Astrophysics Data System (ADS)

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 gravitropism, we sought genetic enhancers of arg1 as a way to identify new gravity signal transducers. Two of these modifiers, named mar1 and mar2, were found to affect genes that encode two subunits of the plastidic outer-membrane protein import complex, TOC75 and TOC132, respectively. mar2 did not affect the ultrastructure of amyloplasts in the statocytes nor did it alter their ability to sediment in response to gravistimulation, suggesting a role for the outer membrane of the amyloplasts in gravity signal transduction (reviewed in Stanga et al., 2009, Plant Signal Behavior 4(10): 1-9). The contribution of TOC132 in gravity signal transduction is being investigated by analyzing the regions of this protein that are needed for the pathway, and investigating the contribution of a putative TOC132-interacting protein in gravity signal transduction. We have also isolated additional putative enhancers of arg1-2 in the hope of identifying new plastid-associated gravity signal transducers, and have initiated a screen for genetic enhancers of mar2 to seek new transducers in the ARG1 branch of the pathway.

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

PubMed

Shi, Jiang-hua; Hao, Xi; Wu, Zhong-chang; Wu, Ping

2009-10-01

Root gravitropism is one of the important factors to determine root architecture. To understand the mechanism underlying root gravitropism, we isolated a rice (Xiushui63) mutant defective in root gravitropism, designated as gls1. Vertical sections of root caps revealed that gls1 mutant displayed normal distribution of amyloplast in the columella cells compared with the wild type. The gls1 mutant was less sensitive to 2,4-dichlorophenoxyacetic acid (2,4-D) and alpha-naphthaleneacetic acid (NAA) than the wild type. Genetic analysis indicated that the phenotype of gls1 mutant was caused by a single recessive mutation, which is mapped in a 255-kb region between RM16253 and CAPS1 on the short arm of chromosome 4.

Mechanical analysis of statolith action in roots and rhizoids

NASA Astrophysics Data System (ADS)

Todd, Paul

1994-08-01

Published observations on the response times following gravistimulation (horizontal positioning) of Chara rhizoids and developing roots of vascular plants with normal and ``starchless'' amyloplasts were reviewed and compared. Statolith motion was found to be consistent with gravitational sedimentation opposed by elastic deformation of an intracellular material. The time required for a statolith to sediment to equilibrium was calculated on the basis of its buoyant density and compared with observed sedimentation times. In the examples chosen, the response time following gravistimulation (from horizontal positioning to the return of downward growth) could be related to the statolith sedimentation time. Such a relationship implies that the transduction step is rapid in comparison with the perception steo following gravistimulation of rhizoids and developing roots.

Mechanical analysis of statolith action in roots and rhizoids.

PubMed

Todd, P

1994-01-01

Published observations on the response times following gravistimulation (horizontal positioning) of Chara rhizoids and developing roots of vascular plants with normal and "starchless" amyloplasts were reviewed and compared. Statolith motion was found to be consistent with gravitational sedimentation opposed by elastic deformation of an intracellular material. The time required for a statolith to sediment to equilibrium was calculated on the basis of its buoyant density and compared with observed sedimentation times. In the examples chosen, the response time following gravistimulation (from horizontal positioning to the return of downward growth) could be related to the statolith sedimentation time. Such a relationship implies that the transduction step is rapid in comparison with the perception step following gravistimulation of rhizoids and developing roots.

Changes in the topography of cellular components in pea root statocytes exposed to high gradient magnetic fields

NASA Astrophysics Data System (ADS)

Belyavskaya, Ninel A.; Polishchuk, Olexandr V.; Kondrachuk, Alexander V.

2005-08-01

High-gradient magnetic field (HGMF) is one of methods, by which gravitropism in plants is studied. The aim of our study was elucidation of HGMF effects on topography of cellular components in root statocytes of 4- day Pisum sativum L. seedlings in comparison to gravistimulation. Under gravistimulation during 5, 30 and 60 min seedlings were rotated 45o; magnetostimulation was carried out along gap between two NdFeB magnets (0.7 T). Morphometric measurements were made from images of whole statocytes, for upper, middle and lower thirds of cells, and proximal and distal halves of cells. Morphometric analysis revealed that HGMF resulted in the redistribution of all cellular components in statocytes. The correlation in the amyloplast distribution between gravistimulation and magnetostimulation was established.

Stereological analysis of gravitropism in protonemata of the moss Ceratodon

NASA Technical Reports Server (NTRS)

Walker, L. M.; Sack, F. D.

1997-01-01

Apical cells of dark-grown protonemata of the moss Cerotodon purpureus are negatively gravitropic. Previous light microscopy has shown that reorientation to the horizontal induces amyloplast sedimentation and redistribution of microtubules. To determine whether other components become redistributed laterally or axially, the apical 35 micrometers of both vertical and horizontal apical cells were compared stereologically using transmission electron microscopy. Reorientation to the horizontal changed the longitudinal distributions of tubular ER, Golgi stacks, and vesicles but not cisternal ER, mitochondria, and plastids. Only plastids showed a statistically significant lateral redistribution after horizontal placement. Qualitative examination of the sedimentation zone showed plastids sedimented close to peripherally located ER with vacuoles displaced above plastids. These results argue against a model where differential tip growth results from a redistribution of Golgi stacks or exocytic vesicles.

How do Arabidopsis Roots Differentiate Hydrotropism from Gravitropism?

PubMed Central

Takahashi, Hideyuki

2007-01-01

Root hydrotropism is a response to moisture gradients, which is considered to be important for drought avoidance. Recent reevaluation of root hydrotropism has emphasised the dominating effect of root gravitropism on it. It has been suggested that amyloplast dynamics inside columella cells and auxin regulation play roles in this interacting mechanism, even though the existence of distinct pathways of two tropisms derived from different stimuli remained unclear. We have recently found two factors that separate the mechanism of hydrotropism from that of gravitropism in Arabidopsis seedling roots. One is the difference in the mode of auxin-mediated growth regulation between two tropisms, and the other is the identification of gene indispensable only for root hydrotropism. Here we summarize the recent progress on root hydrotropism research and discuss the remaining and emerging issues. PMID:19704609

Root gravitropism: a complex response to a simple stimulus?

NASA Technical Reports Server (NTRS)

Rosen, E.; Chen, R.; Masson, P. H.

1999-01-01

Roots avoid depleting their immediate environment of essential nutrients by continuous growth. Root growth is directed by environmental cues, including gravity. Gravity sensing occurs mainly in the columella cells of the root cap. Upon reorientation within the gravity field, the root-cap amyloplasts sediment, generating a physiological signal that promotes the development of a curvature at the root elongation zones. Recent molecular genetic studies in Arabidopsis have allowed the identification of genes that play important roles in root gravitropism. Among them, the ARG1 gene encodes a DnaJ-like protein involved in gravity signal transduction, whereas the AUX1 and AGR1 genes encode proteins involved in polar auxin transport. These studies have important implications for understanding the intra- and inter-cellular signaling processes that underlie root gravitropism.

Gravitropism in leaves of Arabidopsis thaliana (L.) Heynh.

PubMed

Mano, Eriko; Horiguchi, Gorou; Tsukaya, Hirokazu

2006-02-01

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.

Phototropism and gravitropism in lateral roots of Arabidopsis

NASA Technical Reports Server (NTRS)

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

2002-01-01

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.

Phototropism and gravitropism in lateral roots of Arabidopsis.

PubMed

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

2002-01-01

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.

Root gravitropism

NASA Technical Reports Server (NTRS)

Masson, P. H.

1995-01-01

When a plant root is reoriented within the gravity field, it responds by initiating a curvature which eventually results in vertical growth. Gravity sensing occurs primarily in the root tip. It may involve amyloplast sedimentation in the columella cells of the root cap, or the detection of forces exerted by the mass of the protoplast on opposite sides of its cell wall. Gravisensing activates a signal transduction cascade which results in the asymmetric redistribution of auxin and apoplastic Ca2+ across the root tip, with accumulation at the bottom side. The resulting lateral asymmetry in Ca2+ and auxin concentration is probably transmitted to the elongation zone where differential cellular elongation occurs until the tip resumes vertical growth. The Cholodny-Went theory proposes that gravity-induced auxin redistribution across a gravistimulated plant organ is responsible for the gravitropic response. However, recent data indicate that the gravity-induced reorientation is more complex, involving both auxin gradient-dependent and auxin gradient-independent events.

Carotenoid Metabolism in Plants: The Role of Plastids.

PubMed

Sun, Tianhu; Yuan, Hui; Cao, Hongbo; Yazdani, Mohammad; Tadmor, Yaakov; Li, Li

2018-01-08

Carotenoids are indispensable to plants and critical in human diets. Plastids are the organelles for carotenoid biosynthesis and storage in plant cells. They exist in various types, which include proplastids, etioplasts, chloroplasts, amyloplasts, and chromoplasts. These plastids have dramatic differences in their capacity to synthesize and sequester carotenoids. Clearly, plastids play a central role in governing carotenogenic activity, carotenoid stability, and pigment diversity. Understanding of carotenoid metabolism and accumulation in various plastids expands our view on the multifaceted regulation of carotenogenesis and facilitates our efforts toward developing nutrient-enriched food crops. In this review, we provide a comprehensive overview of the impact of various types of plastids on carotenoid biosynthesis and accumulation, and discuss recent advances in our understanding of the regulatory control of carotenogenesis and metabolic engineering of carotenoids in light of plastid types in plants. Copyright © 2017 The Author. Published by Elsevier Inc. All rights reserved.

Genetical approach to gravitropism

NASA Astrophysics Data System (ADS)

Boonsirichai, K.; Chen, R.; Guan, C.; Rosen, E.; Young, L.; Masson, P.

Gravitropism guides the growth of plant organs at a defined angle from the gravity vector. Accordingly, most roots grow downward, undergoing positive gravitropism. Gravity perception by roots appears to involve the sedimentation of amyloplasts within the columella cells of the cap. Amyloplast sedimentation triggers a signal transduction pathway that promotes the development of an auxin gradient across the root tip. This gradient is then transmitted to the elongation zones where it promotes a differential cellular elongation, partly responsible for the development of a root-tip curvature. To better understand the mechanisms involved in gravity signal transduction, we have identified and characterized several Arabidopsis thaliana mutants that show specific defects in root gravitropism. Several of these genes were characterized. ARG1 functions in gravity signal transduction, and encodes a dnaJ-like protein whose structure suggests an interaction with the cytoskeleton. Two other genes encode similar proteins (ARL1 and ARL2) in Arabidopsis. One of them (ARL2) also appears to function in gravity signal transduction. Because loss-of-function mutations in ARG1 result in partial alterations of gravitropism, we were able to identify and characterize two genetic enhancers of arg1-2: mar1-1 and mar2-1. These enhancers increased the gravitropism defect of arg1-2 roots and hypocotyls, and changed its orientation. Hence, MAR1 and MAR2 also appear to function in gravity signal transduction. AGR1, on the other hand, encodes a transmembrane component of the auxin efflux carrier complex involved in polar auxin transport through the elongation zones of Arabidopsis root tips. It belongs to a large gene family, several members of which are expressed in the root cap. Upon gravistimulation, the AGR3 protein appears to quickly relocate within the columella cells, accumulating in membranes at the new physical bottom. Hence, the gravity signal transduction pathway that includes the ARG1, ARL2, MAR1 and MAR2 gene products, appears to control the cellular distribution of auxin efflux carriers in the columella cells of the root cap, thereby controlling the polarity of lateral auxin transport in response to gravistimulation. Work is in progress to identify new proteins that interact genetically or physically with ARG1, ARL2 or AGR1, and characterize their involvement in gravitropism.

Displacement of organelles in plant gravireceptor cells by vibrational forces and ultrasound.

NASA Astrophysics Data System (ADS)

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 reaction to amyloplasts displacement by acoustic forces. Besides investigating the graviperception mechanism, vibrational and acoustic forces can serve as tools for analyzing mechanical properties of cell interior. Practical applications of this technology could include providing directional stimuli for plants in microgravity by low doses of vibrations. Vibrations present on board of spacecraft may have vectorial effects on plants and other organisms, and their influence should be assessed.

Tropism in azalea and lily flowers

NASA Astrophysics Data System (ADS)

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 pistil as long as differential incidence of light was given at its part. Elongation and bending of filament of stamen in lily differed from those of style in several points. After perianth opened, filaments deployed and spread out from the central axis of flower. Distinctive elongation of filament developed at a zone close to anther. It will be discussed how such regulation made by either gravity or light improves the degree of adaptation for those plants during entomophilous pollination.

Distribution of calmodulin in corn seedlings - Immunocytochemical localization in coleoptiles and root apices

NASA Technical Reports Server (NTRS)

Dauwalder, M.; Roux, S. J.

1986-01-01

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.

Growth of cress seedlings and morphogenesis of root Gravisensors under clino-rotation and in unidirectional red or blue light

NASA Astrophysics Data System (ADS)

Rakleviciene, D.; Svegzdiene, D.; Tamulaitis, G.; Zukauskas, A.

2005-08-01

The growth rate and orientation of cress seedlings in response to the direction of illumination under clino- rotation were investigated at the initial stage of intensive hypocotyl elongation. Roots and hypocotyls growing in normal gravity conditions (1 g) and under clino-rotation at 3 rpm were illuminated with red (660 nm) or blue (450 nm) light from light-emitting diodes (LEDs). Unidirectional illumination in the direction opposite to the gravity vector promoted the growth rate of roots. Inhibition of gravitropism by clino-rotation reduced the growth of roots and stimulated the elongation of hypocotyls in both red and blue light. Illumination of roots invoked changes in the formation of gravisensing cells in the columella. Illumination under clino-rotation stimulated root statocyte growth and increased the number of amyloplasts in cells of the 3rd-6th columella rows. Also, an increase in the columella cell area, mainly caused by cell elongation in blue light and by enhanced radial growth in red light, was observed.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Okita, T.W.

1990-12-31

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 andmore » 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.« less

Dynamics of vegetative cytoplasm during generative cell formation and pollen maturation in Arabidopsis thaliana

NASA Technical Reports Server (NTRS)

Kuang, A.; Musgrave, M. E.

1996-01-01

Ultrastructural changes of pollen cytoplasm during generative cell formation and pollen maturation in Arabidopsis thaliana were studied. The pollen cytoplasm develops a complicated ultrastructure and changes dramatically during these stages. Lipid droplets increase after generative cell formation and their organization and distribution change with the developmental stage. Starch grains in amyloplasts increase in number and size during generative and sperm cell formation and decrease at pollen maturity. The shape and membrane system of mitochondria change only slightly. Dictyosomes become very prominent, and numerous associated vesicles are observed during and after sperm cell formation. Endoplasmic reticulum appears extensively as stacks during sperm cell formation. Free and polyribosomes are abundant in the cytoplasm at all developmental stages although they appear denser at certain stages and in some areas. In mature pollen, all organelles are randomly distributed throughout the vegetative cytoplasm and numerous small particles appear. Organization and distribution of storage substances and appearance of these small particles during generative and sperm cell formation and pollen maturation are discussed.

Molecular Mechanisms of Root Gravitropism.

PubMed

Su, Shih-Heng; Gibbs, Nicole M; Jancewicz, Amy L; Masson, Patrick H

2017-09-11

Plant shoots typically grow against the gravity vector to access light, whereas roots grow downward into the soil to take up water and nutrients. These gravitropic responses can be altered by developmental and environmental cues. In this review, we discuss the molecular mechanisms that govern the gravitropism of angiosperm roots, where a physical separation between sites for gravity sensing and curvature response has facilitated discovery. Gravity sensing takes place in the columella cells of the root cap, where sedimentation of starch-filled plastids (amyloplasts) triggers a pathway that results in a relocalization to the lower side of the cell of PIN proteins, which facilitate efflux of the plant hormone auxin efflux. Consequently, auxin accumulates in the lower half of the root, triggering bending of the root tip at the elongation zone. We review our understanding of the molecular mechanisms that control this process in primary roots, and discuss recent insights into the regulation of oblique growth in lateral roots and its impact on root-system architecture and soil exploration. Copyright © 2017 Elsevier Ltd. All rights reserved.

A weak combined magnetic field changes root gravitropism

NASA Astrophysics Data System (ADS)

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.

Analysis of magnetic gradients to study gravitropism.

PubMed

Hasenstein, Karl H; John, Susan; Scherp, Peter; Povinelli, Daniel; Mopper, Susan

2013-01-01

Gravitropism typically is generated by dense particles that respond to gravity. Experimental stimulation by high-gradient magnetic fields provides a new approach to selectively manipulate the gravisensing system. The movement of corn, wheat, and potato starch grains in suspension was examined with videomicroscopy during parabolic flights that generated 20 to 25 s of weightlessness. During weightlessness, a magnetic gradient was generated by inserting a wedge into a uniform, external magnetic field that caused repulsion of starch grains. The resultant velocity of movement was compared with the velocity of sedimentation under 1 g conditions. The high-gradient magnetic fields repelled the starch grains and generated a force of at least 0.6 g. Different wedge shapes significantly affected starch velocity and directionality of movement. Magnetic gradients are able to move diamagnetic compounds under weightless or microgravity conditions and serve as directional stimulus during seed germination in low-gravity environments. Further work can determine whether gravity sensing is based on force or contact between amyloplasts and statocyte membrane system.

Arabidopsis Myosins XI1, XI2, and XIK Are Crucial for Gravity-Induced Bending of Inflorescence Stems

PubMed Central

Talts, Kristiina; Ilau, Birger; Ojangu, Eve-Ly; Tanner, Krista; Peremyslov, Valera V.; Dolja, Valerian V.; Truve, Erkki; Paves, Heiti

2016-01-01

Myosins and actin filaments in the actomyosin system act in concert in regulating cell structure and dynamics and are also assumed to contribute to plant gravitropic response. To investigate the role of the actomyosin system in the inflorescence stem gravitropism, we used single and multiple mutants affecting each of the 17 Arabidopsis myosins of class VIII and XI. We show that class XI but not class VIII myosins are required for stem gravitropism. Simultaneous loss of function of myosins XI1, XI2, and XIK leads to impaired gravitropic bending that is correlated with altered growth, stiffness, and insufficient sedimentation of gravity sensing amyloplasts in stem endodermal cells. The gravitropic defect of the corresponding triple mutant xi1 xi2 xik could be rescued by stable expression of the functional XIK:YFP in the mutant background, indicating a role of class XI myosins in this process. Altogether, our results emphasize the critical contributions of myosins XI in stem gravitropism of Arabidopsis. PMID:28066484

[Microanalytical identification of barium sulphate crystals in statoliths of Chara rhizoids (Ch. fragilis, desv.)].

PubMed

Schröter, K; Läuchli, A; Sievers, A

1975-01-01

In contrast to the statocytes of higher plants, in which amyloplasts function as statoliths, Chara-rhizoids contain statolith vacuoles filled with biocrystallites of BaSO4. This was revealed by qualitative and quantitative electron microprobe analysis, atomic absorption spectrophotometry and selected area electron diffraction. The barium sulphate crystallites are rods which are linearly composed of globular subunits approximately 7 nm in diameter.The electron optical evidence of the crystallites depends on the nature of the fixatives. Best structural preservation was observed after fixation in a buffered solution of glutaraldehyde plus acrolein without addition of heavy metals. OsO4 and particularly KMnO4 partially dissolve the biocrystallites as well as synthetic BaSO4. The crystal solubility must be taken into consideration when micrographs of such small crystallites are interpreted.The fact that BaSO4 is chemically very inert seems to exclude biochemical interactions of the statoliths with other cell components during graviperception. It favours the theory that only the mass of the statoliths is effective.

Allelic variants of the amylose extender mutation of maize demonstrate phenotypic variation in starch structure resulting from modified protein–protein interactions

PubMed Central

Liu, Fushan; Ahmed, Zaheer; Lee, Elizabeth A.; Donner, Elizabeth; Liu, Qiang; Ahmed, Regina; Morell, Matthew K.; Emes, Michael J.; Tetlow, Ian J.

2012-01-01

amylose extender (ae−) starches characteristically have modified starch granule morphology resulting from amylopectin with reduced branch frequency and longer glucan chains in clusters, caused by the loss of activity of the major starch branching enzyme (SBE), which in maize endosperm is SBEIIb. A recent study with ae− maize lacking the SBEIIb protein (termed ae1.1 herein) showed that novel protein–protein interactions between enzymes of starch biosynthesis in the amyloplast could explain the starch phenotype of the ae1.1 mutant. The present study examined an allelic variant of the ae− mutation, ae1.2, which expresses a catalytically inactive form of SBEIIb. The catalytically inactive SBEIIb in ae1.2 lacks a 28 amino acid peptide (Val272–Pro299) and is unable to bind to amylopectin. Analysis of starch from ae1.2 revealed altered granule morphology and physicochemical characteristics distinct from those of the ae1.1 mutant as well as the wild-type, including altered apparent amylose content and gelatinization properties. Starch from ae1.2 had fewer intermediate length glucan chains (degree of polymerization 16–20) than ae1.1. Biochemical analysis of ae1.2 showed that there were differences in the organization and assembly of protein complexes of starch biosynthetic enzymes in comparison with ae1.1 (and wild-type) amyloplasts, which were also reflected in the composition of starch granule-bound proteins. The formation of stromal protein complexes in the wild-type and ae1.2 was strongly enhanced by ATP, and broken by phosphatase treatment, indicating a role for protein phosphorylation in their assembly. Labelling experiments with [γ-32P]ATP showed that the inactive form of SBEIIb in ae1.2 was phosphorylated, both in the monomeric form and in association with starch synthase isoforms. Although the inactive SBEIIb was unable to bind starch directly, it was strongly associated with the starch granule, reinforcing the conclusion that its presence in the granules is a result of physical association with other enzymes of starch synthesis. In addition, an Mn2+-based affinity ligand, specific for phosphoproteins, was used to show that the granule-bound forms of SBEIIb in the wild-type and ae1.2 were phosphorylated, as was the granule-bound form of SBEI found in ae1.2 starch. The data strongly support the hypothesis that the complement of heteromeric complexes of proteins involved in amylopectin synthesis contributes to the fine structure and architecture of the starch granule. PMID:22121198

Changes in root gravitropism, ultrastructure, and calcium balance of pea root statocytes induced by A23187

NASA Astrophysics Data System (ADS)

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 product was found in mitochondrial cristae in contrast to control ones. The presence of the precipitate in other Ca 2 +-sequestered organelles was not determined. The data presented suggest that at the ultrastructural level, the effects of the Ca 2 + ionophore manifested in the loss of polarity in statocytes may be functionally related to systems that regulate the intracellular Ca 2 + homeostasis. It is evident that significant increase in Ca 2 + level in A23187-treated statocytes may cause a disbalance in the gravisensor system and/or calcium signaling and therefore to abolish gravitropism of pea roots.

Influence of Microgravity Environment on Root Growth, Soluble Sugars, and Starch Concentration of Sweetpotato Stem Cuttings

PubMed Central

Mortley, Desmond G.; Bonsi, Conrad K.; Hill, Walter A.; Morris, Carlton E.; Williams, Carol S.; Davis, Ceyla F.; Williams, John W.; Levine, Lanfang H.; Petersen, Barbara V.; Wheeler, Raymond M.

2009-01-01

Because sweetpotato [Ipomoea batatas (L.) Lam.] stem cuttings regenerate very easily and quickly, a study of their early growth and development in microgravity could be useful to an understanding of morphological changes that might occur under such conditions for crops that are propagated vegetatively. An experiment was conducted aboard a U.S. Space Shuttle to investigate the impact of microgravity on root growth, distribution of amyloplasts in the root cells, and on the concentration of soluble sugars and starch in the stems of sweetpotatoes. Twelve stem cuttings of ‘Whatley/Loretan’ sweetpotato (5 cm long) with three to four nodes were grown in each of two plant growth units filled with a nutrient agarose medium impregnated with a half-strength Hoagland solution. One plant growth unit was flown on Space Shuttle Colombia for 5 days, whereas the other remained on the ground as a control. The cuttings were received within 2 h postflight and, along with ground controls, processed in ≈45 min. Adventitious roots were counted, measured, and fixed for electron microscopy and stems frozen for starch and sugar assays. Air samples were collected from the headspace of each plant growth unit for postflight determination of carbon dioxide, oxygen, and ethylene levels. All stem cuttings produced adventitious roots and growth was quite vigorous in both ground-based and flight samples and, except for a slight browning of some root tips in the flight samples, all stem cuttings appeared normal. The roots on the flight cuttings tended to grow in random directions. Also, stem cuttings grown in microgravity had more roots and greater total root length than ground-based controls. Amyloplasts in root cap cells of ground-based controls were evenly sedimented toward one end compared with a more random distribution in the flight samples. The concentration of soluble sugars, glucose, fructose, and sucrose and total starch concentration were all substantially greater in the stems of flight samples than those found in the ground-based samples. Carbon dioxide levels were 50% greater and oxygen marginally lower in the flight plants, whereas ethylene levels were similar and averaged less than 10 nL·L −1. Despite the greater accumulation of carbohydrates in the stems, and greater root growth in the flight cuttings, overall results showed minimal differences in cell development between space flight and ground-based tissues. This suggests that the space flight environment did not adversely impact sweetpotato metabolism and that vegetative cuttings should be an acceptable approach for propagating sweetpotato plants for space applications. PMID:20186286

The effect of the external medium on the gravitropic curvature of rice (Oryza sativa, Poaceae) roots

NASA Technical Reports Server (NTRS)

Staves, M. P.; Wayne, R.; Leopold, A. C.

1997-01-01

The roots of rice seedlings, growing in artificial pond water, exhibit robust gravitropic curvature when placed perpendicular to the vector of gravity. To determine whether the statolith theory (in which intracellular sedimenting particles are responsible for gravity sensing) or the gravitational pressure theory (in which the entire protoplast acts as the gravity sensor) best accounts for gravity sensing in rice roots, we changed the physical properties of the external medium with impermeant solutes and examined the effect on gravitropism. As the density of the external medium is increased, the rate of gravitropic curvature decreases. The decrease in the rate of gravicurvature cannot be attributed to an inhibition of growth, since rice roots grown in 100 Osm/m3 (0.248 MPa) solutions of different densities all support the same root growth rate but inhibit gravicurvature increasingly with increasing density. By contrast, the sedimentation rate of amyloplasts in the columella cells is unaffected by the external density. These results are consistent with the gravitational pressure theory of gravity sensing, but cannot be explained by the statolith theory.

Irradiance-dependent regulation of gravitropism by red light in protonemata of the moss Ceratodon purpureus

NASA Technical Reports Server (NTRS)

Kern, V. D.; Sack, F. D.

1999-01-01

Apical cells of protonemata of the moss Ceratodon purpureus (Hedw.) Brid. are negatively gravitropic in the dark and positively phototropic in red light. Various fluence rates of unilateral red light were tested to determine whether both tropisms operate simultaneously. At irradiances > or = 140 nmol m-2 s-1 no gravitropism could be detected and phototropism predominated, despite the presence of amyloplast sedimentation. Gravitropism occurred at irradiances lower than 140 nmol m-1 s-1 with most cells oriented above the horizontal but not upright. At these low fluence rates, phototropism was indistinct at 1 g but apparent in microgravity, indicating that gravitropism and phototropism compete at 1 g. The frequency of protonemata that were negatively phototropic varied with the fluence rate and the duration of illumination, as well as with the position of the apical cell before illumination. These data show that the fluence rate of red light regulates whether gravitropism is allowed or completely repressed, and that it influences the polarity of phototropism and the extent to which apical cells are aligned in the light path.

Microtubules restrict plastid sedimentation in protonemata of the moss Ceratodon

NASA Technical Reports Server (NTRS)

Schwuchow, J.; Sack, F. D.

1994-01-01

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.

Severely Reduced Gravitropism in Dark-Grown Hypocotyls of a Starch-Deficient Mutant of Nicotiana sylvestris1

PubMed Central

Kiss, John Z.; Sack, Fred D.

1990-01-01

Gravitropism in dark-grown hypocotyls of the wild type was compared with a starch-deficient Nicotiana sylvestris mutant (NS 458) to test the effects of starch deficiency on gravity sensing. In a time course of curvature measured using infrared video, the response of the mutant was greatly reduced compared to the wild type; 72 hours after reorientation, curvature was about 10° for NS 458 and about 70° for wild type. In dishes maintained in a vertical orientation, wild-type hypocotyls were predominantly vertical, whereas NS 458 hypocotyls were severely disoriented with about 5 times more orientational variability than wild type. Since the growth rates were equal for both genotypes and phototropic curvature was only slightly inhibited in NS 458, the mutation probably affects gravity perception rather than differential growth. Our data suggest that starch deficiency reduces gravitropic sensitivity more in dark-grown hypocotyls than in dark- or light-grown roots in this mutant and support the hypothesis that amyloplasts function as statoliths in shoots as well as roots. Images Figure 2 Figure 3 Figure 4 PMID:11537476

Severely reduced gravitropism in dark-grown hypocotyls of a starch-deficient mutant of Nicotiana sylvestris

NASA Technical Reports Server (NTRS)

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

1990-01-01

Gravitropism in dark-grown hypocotyls of the wild type was compared with a starch-deficient Nicotiana sylvestris mutant (NS 458) to test the effects of starch deficiency on gravity sensing. In a time course of curvature measured using infrared video, the response of the mutant was greatly reduced compared to the wild type; 72 hours after reorientation, curvature was about 10 degrees for NS 458 and about 70 degrees for wild type. In dishes maintained in a vertical orientation, wild-type hypocotyls were predominantly vertical, whereas NS 458 hypocotyls were severely disoriented with about 5 times more orientational variability than wild type. Since the growth rates were equal for both genotypes and phototropic curvature was only slightly inhibited in NS 458, the mutation probably affects gravity perception rather than differential growth. Our data suggest that starch deficiency reduces gravitropic sensitivity more in dark-grown hypocotyls than in dark- or light-grown roots in this mutant and support the hypothesis that amyloplasts function as statoliths in shoots as well as roots.

The gravireaction of Ceratodon protonemata treated with gibberellic acid

NASA Astrophysics Data System (ADS)

Chaban, C. I.; Kordyum, E. L.; Demkiv, O. T.; Khorkavtsiv, O. Ya.; Khorkavtsiv, Ya. D.

1999-01-01

Moss protonemata exhibit negative gravitropism and the amyloplasts of the apical cell seem to play a key role in protonemal gravisensitivity. However, the mechanisms of this process are still poorly understood. Previously, we have shown that Ceratodon protonemata grown on agar-medium demonstrated greater gravicurvature than protonemata grown on medium with 11 mM glucose. In this study, we have examined whether gibberellic acid (GA), which promotes α-amylase expression, influences graviresponse of C. purpureus protonemata (strains WT-4 and WT-U) and how this event interacts with exogenous soluble sugars. After gravistimulation the WT-4 strain curved about twice as fast as the WT-U strain. However, responses of both strains to added substances were similar. High concentration of glucose (0.11 M) caused a decrease in protonema curvature, while the same concentration of sucrose did not significantly change the angles of curvature compared with controls. GA at 0.1 mM and higher concentrations inhibited gravitropism, and caused some apical cells to swell. The possible involvement of the carbohydrates in gravitropism is discussed.

Ultrastructural study on dynamics of plastids and mitochondria during microgametogenesis in watermelon.

PubMed

Liu, Lin

2012-02-01

Dynamics of plastids and mitochondria during microgametogenesis in watermelon were examined by means of transmission electron microscopy. Plastids are present as proplastids in the microspore and as amyloplasts in the vegetative cell of the bicellular pollen grain, whereas the generative cell is completely devoid of plastids, suggesting that microspore plastids are excluded from the generative cell during the microspore mitotic division. Therefore, watermelon is classified as Lycopersicon type, where plastids exclusion from the generative cell leads to purely maternal plastid inheritance. Mitochondria in the generative cell show noticeable alterations in size and cristae during microgametogenesis. The diameter of mitochondria is about 0.5 μm in the newly born generative cell, while only about 0.16 μm in the spindle-shaped generative cell. Numerous cristae are present in mitochondria in the spherical generative cell, but, in contrast, mere two or three cristae retain in the spindle-shaped generative cell in the mature pollen grain. In conclusion, the size and cristae number of mitochondria in the generative cell are reduced significantly during microgametogenesis in watermelon. Copyright © 2011 Elsevier Ltd. All rights reserved.

Space Magnets Attracting Interest on Earth: Applications of Physical and Biological Techniques In the Study of Gravisensing and Response System of Plants

NASA Technical Reports Server (NTRS)

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

2002-01-01

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.

Signal perception, transduction, and response in gravity resistance. Another graviresponse in plants

NASA Astrophysics Data System (ADS)

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.

A functional TOC complex contributes to gravity signal transduction in Arabidopsis

PubMed Central

Strohm, Allison K.; Barrett-Wilt, Greg A.; Masson, Patrick H.

2014-01-01

Although plastid sedimentation has long been recognized as important for a plant's perception of gravity, it was recently shown that plastids play an additional function in gravitropism. The Translocon at the Outer envelope membrane of Chloroplasts (TOC) complex transports nuclear-encoded proteins into plastids, and a receptor of this complex, Toc132, was previously hypothesized to contribute to gravitropism either by directly functioning as a gravity signal transducer or by indirectly mediating the plastid localization of a gravity signal transducer. Here we show that mutations in multiple genes encoding TOC complex components affect gravitropism in a genetically sensitized background and that the cytoplasmic acidic domain of Toc132 is not required for its involvement in this process. Furthermore, mutations in TOC132 enhance the gravitropic defect of a mutant whose amyloplasts lack starch. Finally, we show that the levels of several nuclear-encoded root proteins are altered in toc132 mutants. These data suggest that the TOC complex indirectly mediates gravity signal transduction in Arabidopsis and support the idea that plastids are involved in gravitropism not only through their ability to sediment but also as part of the signal transduction mechanism. PMID:24795735

A functional TOC complex contributes to gravity signal transduction in Arabidopsis.

PubMed

Strohm, Allison K; Barrett-Wilt, Greg A; Masson, Patrick H

2014-01-01

Although plastid sedimentation has long been recognized as important for a plant's perception of gravity, it was recently shown that plastids play an additional function in gravitropism. The Translocon at the Outer envelope membrane of Chloroplasts (TOC) complex transports nuclear-encoded proteins into plastids, and a receptor of this complex, Toc132, was previously hypothesized to contribute to gravitropism either by directly functioning as a gravity signal transducer or by indirectly mediating the plastid localization of a gravity signal transducer. Here we show that mutations in multiple genes encoding TOC complex components affect gravitropism in a genetically sensitized background and that the cytoplasmic acidic domain of Toc132 is not required for its involvement in this process. Furthermore, mutations in TOC132 enhance the gravitropic defect of a mutant whose amyloplasts lack starch. Finally, we show that the levels of several nuclear-encoded root proteins are altered in toc132 mutants. These data suggest that the TOC complex indirectly mediates gravity signal transduction in Arabidopsis and support the idea that plastids are involved in gravitropism not only through their ability to sediment but also as part of the signal transduction mechanism.

Complex physiological and molecular processes underlying root gravitropism

NASA Technical Reports Server (NTRS)

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

2002-01-01

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.

The possible involvement of root-cap mucilage in gravitropism and calcium movement across root tips of Allium cepa L

NASA Technical Reports Server (NTRS)

Moore, R.; Fondren, W. M.

1986-01-01

Roots of Allium cepa L. grown in aerated water elongate rapidly, but are not graviresponsive. These roots (1) possess extensive columella tissues comprised of cells containing numerous sedimented amyloplasts, (2) lack mucilage on their tips, and (3) are characterized by a weakly polar movement of calcium (Ca) across their tips. Placing roots in humid air correlates positively with the (1) onset of gravicurvature, (2) appearance of mucilage on tips of the roots, and (3) onset of the ability to transport Ca polarly to the lower side of the root tip. Gravicurvature of roots previously submerged in aerated water is more rapid when roots are oriented vertically for 1-2 h in humid air prior to being oriented horizontally. The more rapid gravicurvature of these roots correlates positively with the accumulation of mucilage at the tips of roots during the time the roots are oriented vertically. Therefore, the onset of gravicurvature and the ability of roots to transport Ca to the lower sides of their tips correlate positively with the presence of mucilage at their tips. These results suggest that mucilage may be important for the transport of Ca across root caps.

How cereal grass shoots perceive and respond to gravity

NASA Technical Reports Server (NTRS)

Kaufman, P. B.; Brock, T. G.; Song, I.; Rho, Y. B.; Ghosheh, N. S.

1987-01-01

The leaf-sheath pulvinus of grasses presents a unique system for studying gravitropism, primarily because of its differences from other organs. The mature pulvinus is a discrete organ specialized for gravitropism: it is nongrowing in the absence of gravistimulation and capable of displaying a graviresponse independent of the rest of the plant. In this paper we present a model for gravitropism in pulvini based on recent findings from studies on the mechanisms of graviperception and graviresponse. According to this model, amyloplasts play an essential role in perceiving a change in the orientation of the pulvinus. The perception of this reorientation leads to the enhanced synthesis and release from conjugate of the auxin IAA, and the increased conjugation of gibberellin, on a localized basis. Because there is a graded growth promotion across the gravistimulated pulvinus, it is suggested that the observed hormonal asymmetry is actually an indication of a linear gradient of hormone concentration, as well as hormone response, across the pulvinus. It is further suggested that the linear gradient of hormone concentration may be predominantly the result of local changes in hormone level, rather than a product of hormonal movement into or across the pulvinus.

Critical and speculative review of the roles of multi-protein complexes in starch biosynthesis in cereals.

PubMed

Crofts, Naoko; Nakamura, Yasunori; Fujita, Naoko

2017-09-01

Starch accounts for the majority of edible carbohydrate resources generated through photosynthesis. Amylopectin is the major component of starch and is one of highest-molecular-weight biopolymers. Rapid and systematic synthesis of frequently branched hydro-insoluble amylopectin and efficient accumulation into amyloplasts of cereal endosperm is crucial. The functions of multiple starch biosynthetic enzymes, including elongation, branching, and debranching enzymes, must be temporally and spatially coordinated. Accordingly, direct evidence of protein-protein interactions of starch biosynthetic enzymes were first discovered in developing wheat endosperm in 2004, and they have since been shown in the developing seeds of other cereals. This review article describes structural characteristics of starches as well as similarities and differences in protein complex formation among different plant species and among mutant plants that are deficient in specific starch biosynthetic enzymes. In addition, evidence for protein complexes that are involved in the initiation stages of starch biosynthesis is summarized. Finally, we discuss the significance of protein complexes and describe new methods that may elucidate the mechanisms and roles of starch biosynthetic enzyme complexes. Copyright © 2017 Elsevier B.V. All rights reserved.

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

PubMed

Muratov, Alexander; Baulin, Vladimir A

2015-12-01

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. Copyright © 2015 Elsevier B.V. All rights reserved.

Participation of IAA in transduction of gravistimulus in apical cells of moss protonema

NASA Astrophysics Data System (ADS)

Oksyniuk, U. A.; Khorkavtsiv, O. Y.; Lesniak, Y. I.

Growth movements of vascular plant axis organs -- photo-, gravi- and other tropisms -- are tightly connected with IAA transport (Hertel, 1983; Medvedev, 1996; Kiss, 2000). Moss protonema synthesizes IAA (indole-3-acetic acid) and transports it basipetally favouring growth and differentiation of caulonema (Bopp, 1979; Rose, Bopp, 1983; Rose et al., 1983). We aimed at studying the role of IAA in moss protonema gravitropism using exogenous IAA, 1-NAA (1-naphthaleneacetic acid), 2,4D (2,4-dichlorophenoxyacetic acid) and inhibitors of polar IAA transport -- phytotropins NPA (N-1-naphthylphthalamic acid) and TIBA (2,3,5-triiodobenzoic acid). Six-day gravitropic protonema of Ceratodon purpureus and Pohlia nutans were taken for experiments. Auxin and phytotropins solutions were laid on protonema mats the latters being kept in solutions for 30 min. Then the surplus of solutions were poured off and Petri dishes were placed vertically for 6 h. 20 μ M of IAA and of other synthetic auxins did not significantly influence the angle of protonema gravity bending, 40 μ M of the agents, howewer, reduced the per cent of apical cells bendings and their angles. The most expressed influence on the angles of bending had the inhibitors of polar IAA transport -- NPA. 0,1 -- 3,0 μ M of this phytotropin did not change the form of apical cell, did not disturb the general distribution of amyloplasts and did not significantly lower the per cent and the value of gravity bending angle, though 10 μ M of the phytotropin - inhibited gravity bending. The mixture of 1-NAA and NPA having been added into the medium the influence of NPA was lowered and gravitropic growth renewed in course of time. 10 μ M of other phytopropin TIBA also inhibited gravitropism of Ceratodon purpureus and Pohlia nutans protonema. The analysis of basipetal transport of IAA in moss rhizoids and protonema may indicate the availability of special IAA transport in these structures (Bopp, Cerier, 1988). On the basis of the carried out experiments it can be suggested that high concentrations of IAA and 1-NAA result in surplus of IAA cells led, probably, to a destruction of the apical-basal gradient in cells. Our results testify that NPA inhibits the gravitropism stronger than the growth of protonema. The peculiarity of moss protonema is that the growth orientation change is a result of a transference of growth zone in the apical cell dome caused by amyloplasts sedimentation inducing lateral asymmetry of Ca2+ and apical-basal IAA flow what in its turn manifests itself in distribution of IAA and/or Ca2+ channels in apical cell dome plasma membrane ( Schwuchow et al., 2001). The transport of IAA in apical cells, probably, functionally polarizes it and just that polarizing function is dominant in cells with tip growth.

New moss species with gravitropic protonemata

NASA Astrophysics Data System (ADS)

Lobachevska, O. V.

Gravitropism of 30 moss species was analysed at different stages of development: germination of spores, protonemata, gametophore and sporophyte formation. Spores were sowed in sterile conditions from the closed capsules on 1 % bactoagar with 0,2 % glucose and cultivated in the dark in vertically oriented petri dishes. In the same conditions fragments of protonemata and gametophores were grown being transferred aseptically from sterile cultures of spores germinated in controled light conditions. To assess gravity sensitivity the dishes were kept upright for 7 10 days in darkness and then 90o turned. After 20 h gravistimulation the angles of apical cell gravity bending were determined. The amount of amyloplasts and their distribution during growth and spatial reorientation of sporophytes selected from nature samples on different stages of species-specific capsule formation were analyzed after JK2J staining. The gravitropic sensing was established in 7 new moss species only. The general traits of all such species were the ark-like cygneous seta bending and inclined, to pendulous, capsules. JK2J staining of young isolated sporophytes has shown, that twisting and bending of seta as well as the spatial capsule reorientation result from the changes of distribution of amyloplasts in the direction of gravitropic growth or caused by their lateral sedimentation. In the dark protonemata of investigated mosses grew upwards on agar surface giving rise to bundles of negatively gravitropic stolons in 7-10 days. During germination at first negatively gravitropic primary chloronema and then positively gravitropic primary rizoid appeared. In 3 days, however, the growth of all primary filaments was negatively gravitropic. In Dicranella cerviculata majority of primary filaments were negatively gravitropic from the very beginning. After 20 h gravistimulation of protonemata of different moss species the following mean values of gravity bending (degrees) were established: Leptobryum pyriforme (65,7± 1,2); Bryum caespiticium (37,2± 1,7); B. argenteum (41,1± 4,3); B. intermedium (33,3± 3,4); Dicranella cerviculata (65,9± 1,8), D. heteromalla (36,2± 1,7) and D. varia (36,7± 1,6). The species differ between themselves in the amount and in the formation rate of gravitropic filaments, as well as in gravitropic sensing at different stages of development. The greatest number of gravitropic filaments and the highest rate of bending were seen in protonemata of L. pyriforme and D. cerviculata. Gravitropic sensing in protonemata of L. pyriforme appeared to be higher in than in gametophores (30,2± 1,9), whereas in B. caespiticium more gravity sensitive were in gametophores (59,9± 1,5), forming lot of gravitropic filaments along all shoot (33,8± 3,7). In the other investigated species gravitropic filaments formed polarily on apex and in basis of shoot, in number not greater than 10. This research was supported by NASA grant NN-09 (R).

Arabidopsis ANGULATA10 is required for thylakoid biogenesis and mesophyll development

PubMed Central

Micol, José Luis

2014-01-01

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. PMID:24663344

Distribution of calmodulin in pea seedlings: immunocytochemical localization in plumules and root apices

NASA Technical Reports Server (NTRS)

Dauwalder, M.; Roux, S. J.; Hardison, L.

1986-01-01

Immunofluorescence techniques have been used to study the distribution of calmodulin in several tissues in young etiolated pea (Pisum sativum L.) seedlings. A fairly uniform staining was seen in the nucleoplasm and background cytoplasm of most cell types. Cell walls and nucleoli were not stained. In addition, patterned staining reactions were seen in many cells. In cells of the plumule, punctate staining of the cytoplasm was common, and in part this stain appeared to be associated with the plastids. A very distinctive staining of amyloplasts was seen in the columella of the root cap. Staining associated with cytoskeletal elements could be shown in division stages. By metaphase, staining of the spindle region was quite evident. In epidermal cells of the stem and along the underside of the leaf there was an intense staining of the vacuolar contents. Guard cells lacked this vacuolar stain. Vacuolar staining was sometimes seen in cells of the stele, but the most distinctive pattern in the stele was associated with young conducting cells of the xylem. These staining patterns are consistent with the idea that the interactions of plastids and the cytoskeletal may be one of the Ca(2+)-mediated steps in the response of plants to environmental stimuli. Nuclear functions may also be controlled, at least in part, by Ca2+.

Characterization of a novel gravitropic mutant of morning glory, weeping2

NASA Astrophysics Data System (ADS)

Kitazawa, Daisuke; Miyazawa, Yutaka; Fujii, Nobuharu; Nitasaka, Eiji; Takahashi, Hideyuki

2008-09-01

In higher plants, gravity is a major environmental cue that governs growth orientation, a phenomenon termed gravitropism. It has been suggested that gravity also affects other aspects of morphogenesis, such as circumnutation and winding movements. Previously, we showed that these aspects of plant growth morphology require amyloplast sedimentation inside gravisensing endodermal cells. However, the molecular mechanism of the graviresponse and its relationship to circumnutation and winding remains obscure. Here, we have characterized a novel shoot gravitropic mutant of morning glory, weeping2 ( we2). In the we2 mutant, the gravitropic response of the stem was absent, and hypocotyls exhibited a severely reduced gravitropic response, whereas roots showed normal gravitropism. In agreement with our previous studies, we found that we2 mutant has defects in shoot circumnutation and winding. Histological analysis showed that we2 mutant forms abnormal endodermal cells. We identified a mutation in the morning glory homolog of SHORT-ROOT ( PnSHR1) that was genetically linked to the agravitropic phenotype of we2 mutant, and which may underlie the abnormal differentiation of endodermal cells in this plant. These results suggest that the phenotype of we2 mutant is due to a mutation of PnSHR1, and that PnSHR1 regulates gravimorphogenesis, including circumnutation and winding movements, in morning glory.

NADPH Thioredoxin Reductase C Is Localized in Plastids of Photosynthetic and Nonphotosynthetic Tissues and Is Involved in Lateral Root Formation in Arabidopsis[W

PubMed Central

Kirchsteiger, Kerstin; Ferrández, Julia; Pascual, María Belén; González, Maricruz; Cejudo, Francisco Javier

2012-01-01

Plastids are organelles present in photosynthetic and nonphotosynthetic plant tissues. While it is well known that thioredoxin-dependent redox regulation is essential for leaf chloroplast function, little is known of the redox regulation in plastids of nonphotosynthetic tissues, which cannot use light as a direct source of reducing power. Thus, the question remains whether redox regulation operates in nonphotosynthetic plastid function and how it is integrated with chloroplasts for plant growth. Here, we show that NADPH-thioredoxin reductase C (NTRC), previously reported as exclusive to green tissues, is also expressed in nonphotosynthetic tissues of Arabidopsis thaliana, where it is localized to plastids. Moreover, we show that NTRC is involved in maintaining the redox homeostasis of plastids also in nonphotosynthetic organs. To test the relationship between plastids of photosynthetic and nonphotosynthetic tissues, transgenic plants were obtained with redox homeostasis restituted exclusively in leaves or in roots, through the expression of NTRC under the control of organ-specific promoters in the ntrc mutant. Our results show that fully functional root amyloplasts are not sufficient for root, or leaf, growth, but fully functional chloroplasts are necessary and sufficient to support wild-type rates of root growth and lateral root formation. PMID:22505729

Isolation of New Gravitropic Mutants under Hypergravity Conditions.

PubMed

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

2016-01-01

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 .

Isolation of New Gravitropic Mutants under Hypergravity Conditions

PubMed Central

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

2016-01-01

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. PMID:27746791

Comparison of the ultrastructure of conventionally fixed and high pressure frozen/freeze substituted root tips of Nicotiana and Arabidopsis

NASA Technical Reports Server (NTRS)

Kiss, J. Z.; Giddings, T. H. Jr; Staehelin, L. A.; Sack, F. D.

1990-01-01

To circumvent the limitations of chemical fixation (CF) and to gain more reliable structural information about higher plant tissues, we have cryofixed root tips of Nicotiana and Arabidopsis by high pressure freezing (HPF). Whereas other freezing techniques preserve tissue to a relatively shallow depth, HPF in conjunction with freeze substitution (FS) resulted in excellent preservation of entire root tips. Compared to CF, in tissue prepared by HPF/FS: (1) the plasmalemma and all internal membranes were much smoother and often coated on the cytoplasmic side by a thin layer of stained material, (2) the plasmalemma was appressed to the cell wall, (3) organelle profiles were rounder, (4) the cytoplasmic, mitochondrial, and amyloplast matrices were denser, (5) vacuoles contained electron dense material, (6) microtubules appeared to be more numerous and straighter, with crossbridges observed between them, (7) cisternae of endoplasmic reticulum (ER) were wider and filled with material, (8) Golgi intercisternal elements were more clearly resolved and were observed between both Golgi vesicles and cisternae, and (9) larger vesicles were associated with Golgi stacks. This study demonstrates that HPF/FS can be used to successfully preserve the ultrastructure of relatively large plant tissues without the use of intracellular cryoprotectants.

Automorphogenesis and gravitropism of plant seedlings grown under microgravity conditions

NASA Astrophysics Data System (ADS)

Hoson, T.; Saiki, M.; Kamisaka, S.; Yamashita, M.

Plant seedlings exhibit automorphogenesis on clinostats. The occurrence of automorphogenesis was confirmed under microgravity in Space Shuttle STS-95 flight. Rice coleoptiles showed an inclination toward the caryopsis in the basal region and a spontaneous curvature in the same adaxial direction in the elongating region both on a three-dimensional (3-D) clinostat and in space. Both rice roots and Arabidopsis hypocotyls also showed a similar morphology in space and on the 3-D clinostat. In rice coleoptiles, the mechanisms inducing such an automorphic curvature were studied. The faster-expanding convex side of rice coleoptiles showed a higher extensibility of the cell wall than the opposite side. Also, in the convex side, the cell wall thickness was smaller, the turnover of the matrix polysaccharides was more active, and the microtubules oriented more transversely than the concave side, and these differences appear to be causes of the curvature. When rice coleoptiles grown on the 3-D clinostat were placed horizontally, the gravitropic curvature was delayed as compared with control coleoptiles. In clinostatted coleoptiles, the corresponding suppression of the amyloplast development was also observed. Similar results were obtained in Arabidopsis hypocotyls. Thus, the induction of automorphogenesis and a concomitant decrease in graviresponsiveness occurred in plant shoots grown under microgravity conditions.

Β-Amylase from Starchless Seeds of Trigonella Foenum-Graecum and Its Localization in Germinating Seeds

PubMed Central

Srivastava, Garima; Kayastha, Arvind M.

2014-01-01

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

Plastid sedimentation kinetics in roots of wild-type and starch-deficient mutants of Arabidopsis

NASA Technical Reports Server (NTRS)

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

1999-01-01

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.

Phytotoxicity of cobalt ions on the duckweed Lemna minor - Morphology, ion uptake, and starch accumulation.

PubMed

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

2015-07-01

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. Copyright © 2015 Elsevier Ltd. All rights reserved.

NADPH thioredoxin reductase C is localized in plastids of photosynthetic and nonphotosynthetic tissues and is involved in lateral root formation in Arabidopsis.

PubMed

Kirchsteiger, Kerstin; Ferrández, Julia; Pascual, María Belén; González, Maricruz; Cejudo, Francisco Javier

2012-04-01

Plastids are organelles present in photosynthetic and nonphotosynthetic plant tissues. While it is well known that thioredoxin-dependent redox regulation is essential for leaf chloroplast function, little is known of the redox regulation in plastids of nonphotosynthetic tissues, which cannot use light as a direct source of reducing power. Thus, the question remains whether redox regulation operates in nonphotosynthetic plastid function and how it is integrated with chloroplasts for plant growth. Here, we show that NADPH-thioredoxin reductase C (NTRC), previously reported as exclusive to green tissues, is also expressed in nonphotosynthetic tissues of Arabidopsis thaliana, where it is localized to plastids. Moreover, we show that NTRC is involved in maintaining the redox homeostasis of plastids also in nonphotosynthetic organs. To test the relationship between plastids of photosynthetic and nonphotosynthetic tissues, transgenic plants were obtained with redox homeostasis restituted exclusively in leaves or in roots, through the expression of NTRC under the control of organ-specific promoters in the ntrc mutant. Our results show that fully functional root amyloplasts are not sufficient for root, or leaf, growth, but fully functional chloroplasts are necessary and sufficient to support wild-type rates of root growth and lateral root formation.

Noise amplification of plant gravisensing

NASA Astrophysics Data System (ADS)

Ma, Zhong; Hasenstein, Karl H.

A common problem in sensory physiology is the detection of weak signals, such as that produced by the repositioning of statoliths (amyloplasts) in gravisensing cells, from a noisy background. As in other studied biological mechanosensory systems, it is conceivable that the gravisensing process may be amplified by stochastic resonance and nonlinear noise-assisted effects. We therefore investigated the possible dependency of gravisensing on vibrational or thermodynamic noise by examining the effect of external oscillation and temperature on the rate of gravitropic curvature in flax (Linum usitatissimum L.) roots. Roots were oscillated for 15 min prior to or during gravistimulation, either parallel or perpendicular to the root axis. The effect of oscillation was dependent on its direction as well as frequency and amplitude. Initial curvature was most effectively enhanced by vertical oscillations of 5 Hz and 0.5 mm amplitude prior to reorientation. Vertically oscillated roots reached half-maximal curvature 32 min after reorientation, about 18 min earlier than non-oscillated roots. The enhancing effect of vibration on curvature subsided with a half-time of about 20 min. The temperature dependency of the graviresponse indicated that thermodynamic noise also impacted gravity perception. For vibrations and temperature studies, the presentation times decreased almost 6-fold. Our data indicate that gravisensing may depend on or be enhanced by thermodynamic or mechanical noise.

Magnetophoretic Induction of Root Curvature

NASA Technical Reports Server (NTRS)

Hasenstein, Karl H.

1997-01-01

The last year of the grant period concerned the consolidation of previous experiments to ascertain that the theoretical premise apply not just to root but also to shoots. In addition, we verified that high gradient magnetic fields do not interfere with regular cellular activities. Previous results have established that: (1) intracellular magnetophoresis is possible; and (2) HGMF lead to root curvature. In order to investigate whether HGMF affect the assembly and/or organization of structural proteins, we examined the arrangement of microtubules in roots exposed to HGMF. The cytoskeletal investigations were performed with fomaldehyde-fixed, nonembedded tissue segments that were cut with a vibratome. Microtubules (MTs) were stained with rat anti-yeast tubulin (YOL 1/34) and DTAF-labeled antibody against rat IgG. Microfilaments (MFs) were visualized by incubation in rhodamine-labeled phalloidin. The distribution and arrangement of both components of the cytoskeleton were examined with a confocal microscope. Measurements of growth rates and graviresponse were done using a video-digitizer. Since HGMF repel diamagnetic substances including starch-filled amyloplasts and most The second aspect of the work includes studies of the effect of cytoskeletal inhibitors on MTs and MFs. The analysis of the effect of micotubular inhibitors on the auxin transport in roots showed that there is very little effect of MT-depolymerizing or stabilizing drugs on auxin transport. This is in line with observations that application of such drugs is not immediately affecting the graviresponsiveness of roots.

Shoot circumnutation and winding movements require gravisensing cells-mediated graviresponse

NASA Astrophysics Data System (ADS)

Kitazawa, D.; Miyazawa, Y.; Fujii, N.; Nitasaka, E.; Takahashi, H.

The stationary nature of plants distinguishes them from other organisms Because of this unique nature higher plants have evolved various mechanisms for responding to environmental cues enabling them to utilize limited resources or to escape from environmental stresses One of the most important mechanisms that plants have acquired is the ability to sense gravity and to use it as a basis for governing their growth orientation a process known as gravitropism In addition to gravitropism oscillatory movement termed circumnutation and winding movement of climbing plants are also important mechanisms that allow plants to elevate their apical meristems to higher positions and these movements are hypothesized to be gravity-related However the relationship between the graviresponse and these movements has not been clarified To verify the necessity of the graviresponse in these movements we used a climbing plant namely Japanese morning glory as a model plant for it has winding growth that allow us to approach the above-mentioned issues We analyzed two distinct mutant lines of morning glory weeping1 and weeping2 both of which have loss of shoot gravitropism Histological characterization revealed that weeping1 has defect in development of gravisensing cells i e endodermis whereas weeping2 has normally developed endodermis with their amyloplasts sediment in response to gravity These observations suggest that these mutants have defect at a different point in the process of the graviresponse cascade Moreover

Transcriptional and Hormonal Regulation of Gravitropism of Woody Stems in Populus[OPEN

PubMed Central

Gerttula, Suzanne; Zinkgraf, Matthew; Lewis, Daniel R.; Brumer, Harry; Hart, Foster; Filkov, Vladimir

2015-01-01

Angiosperm trees reorient their woody stems by asymmetrically producing a specialized xylem tissue, tension wood, which exerts a strong contractile force resulting in negative gravitropism of the stem. Here, we show, in Populus trees, that initial gravity perception and response occurs in specialized cells through sedimentation of starch-filled amyloplasts and relocalization of the auxin transport protein, PIN3. Gibberellic acid treatment stimulates the rate of tension wood formation and gravibending and enhances tissue-specific expression of an auxin-responsive reporter. Gravibending, maturation of contractile fibers, and gibberellic acid (GA) stimulation of tension wood formation are all sensitive to transcript levels of the Class I KNOX homeodomain transcription factor-encoding gene ARBORKNOX2 (ARK2). We generated genome-wide transcriptomes for trees in which gene expression was perturbed by gravistimulation, GA treatment, and modulation of ARK2 expression. These data were employed in computational analyses to model the transcriptional networks underlying wood formation, including identification and dissection of gene coexpression modules associated with wood phenotypes, GA response, and ARK2 binding to genes within modules. We propose a model for gravitropism in the woody stem in which the peripheral location of PIN3-expressing cells relative to the cambium results in auxin transport toward the cambium in the top of the stem, triggering tension wood formation, while transport away from the cambium in the bottom of the stem triggers opposite wood formation. PMID:26410302

Proteomic alterations in root tips of Arabidopsis thaliana seedlings under altered gravity conditions

NASA Astrophysics Data System (ADS)

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

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

PubMed Central

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

2015-01-01

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

New insights into root gravitropic signalling

PubMed Central

Sato, Ethel Mendocilla; Hijazi, Hussein; Bennett, Malcolm J.; Vissenberg, Kris; Swarup, Ranjan

2015-01-01

An important feature of plants is the ability to adapt their growth towards or away from external stimuli such as light, water, temperature, and gravity. These responsive plant growth movements are called tropisms and they contribute to the plant’s survival and reproduction. Roots modulate their growth towards gravity to exploit the soil for water and nutrient uptake, and to provide anchorage. The physiological process of root gravitropism comprises gravity perception, signal transmission, growth response, and the re-establishment of normal growth. Gravity perception is best explained by the starch–statolith hypothesis that states that dense starch-filled amyloplasts or statoliths within columella cells sediment in the direction of gravity, resulting in the generation of a signal that causes asymmetric growth. Though little is known about the gravity receptor(s), the role of auxin linking gravity sensing to the response is well established. Auxin influx and efflux carriers facilitate creation of a differential auxin gradient between the upper and lower side of gravistimulated roots. This asymmetric auxin gradient causes differential growth responses in the graviresponding tissue of the elongation zone, leading to root curvature. Cell biological and mathematical modelling approaches suggest that the root gravitropic response begins within minutes of a gravity stimulus, triggering genomic and non-genomic responses. This review discusses recent advances in our understanding of root gravitropism in Arabidopsis thaliana and identifies current challenges and future perspectives. PMID:25547917

Product stability and sequestration mechanisms in Solanum tuberosum engineered to biosynthesize high value ketocarotenoids.

PubMed

Mortimer, Cara L; Misawa, Norihiko; Ducreux, Laurence; Campbell, Raymond; Bramley, Peter M; Taylor, Mark; Fraser, Paul D

2016-01-01

To produce commercially valuable ketocarotenoids in Solanum tuberosum, the 4, 4' β-oxygenase (crtW) and 3, 3' β-hydroxylase (crtZ) genes from Brevundimonas spp. have been expressed in the plant host under constitutive transcriptional control. The CRTW and CRTZ enzymes are capable of modifying endogenous plant carotenoids to form a range of hydroxylated and ketolated derivatives. The host (cv. Désirée) produced significant levels of nonendogenous carotenoid products in all tissues, but at the apparent expense of the economically critical metabolite, starch. Carotenoid levels increased in both wild-type and transgenic tubers following cold storage; however, stability during heat processing varied between compounds. Subcellular fractionation of leaf tissues revealed the presence of ketocarotenoids in thylakoid membranes, but not predominantly in the photosynthetic complexes. A dramatic increase in the carotenoid content of plastoglobuli was determined. These findings were corroborated by microscopic analysis of chloroplasts. In tuber tissues, esterified carotenoids, representing 13% of the total pigment found in wild-type extracts, were sequestered in plastoglobuli. In the transgenic tubers, this proportion increased to 45%, with esterified nonendogenous carotenoids in place of endogenous compounds. Conversely, nonesterified carotenoids in both wild-type and transgenic tuber tissues were associated with amyloplast membranes and starch granules. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

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

PubMed

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

2011-11-01

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.

Gravitropism in Higher Plant Shoots 1

PubMed Central

Sliwinski, Julianne E.; Salisbury, Frank B.

1984-01-01

Cross and longitudinal sections were prepared for light microscopy from vertical control plants (Xanthium strumarium L. Chicago strain), free-bending horizontal stems, plants restrained 48 hours in a horizontal position, and plants restrained 48 hours and then released, bending immediately about 130°. Top cells of free-bending stems shrink or elongate little; bottom cells continue to elongate. In restrained stems, bottom cells elongate some and increase in diameter; top cells elongate about as much but decrease in diameter. Upon release, bottom cells elongate more and decrease in diameter, while top cells shorten and increase in diameter, accounting for the bend. During restraint, bottom cells take up water while tissue pressures increase; top cells fail to take up water although tissue pressures are decreasing. Settling of amyloplasts was observed in cells of the starch sheath. Removal of different amounts of stem (Xanthium; Lycopersicon esculentum Miller, cv Bonny Best; Ricinus communis L. cv Yolo Wonder) showed that perception of gravity occurs in the bending (elongation) zone, although bending of fourth and fifth internodes from the top was less than in uncut controls. Uniform application of 1% indoleacetic acid in lanolin to cut stem surfaces partially restored bending. Reversing the gradient in tension/compression in horizontal stems (top under compression, bottom under tension) did not affect gravitropic bending. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 10 PMID:16663939

Changes in plastid proteome and structure in arbuscular mycorrhizal roots display a nutrient starvation signature.

PubMed

Daher, Zeina; Recorbet, Ghislaine; Solymosi, Katalin; Wienkoop, Stefanie; Mounier, Arnaud; Morandi, Dominique; Lherminier, Jeannine; Wipf, Daniel; Dumas-Gaudot, Eliane; Schoefs, Benoît

2017-01-01

During arbuscular mycorrhizal symbiosis, arbuscule-containing root cortex cells display a proliferation of plastids, a feature usually ascribed to an increased plant anabolism despite the lack of studies focusing on purified root plastids. In this study, we investigated mycorrhiza-induced changes in plastidic pathways by performing a label-free comparative subcellular quantitative proteomic analysis targeted on plastid-enriched fractions isolated from Medicago truncatula roots, coupled to a cytological analysis of plastid structure. We identified 490 root plastid protein candidates, among which 79 changed in abundance upon mycorrhization, as inferred from spectral counting. According to cross-species sequence homology searches, the mycorrhiza-responsive proteome was enriched in proteins experimentally localized in thylakoids, whereas it was depleted of proteins ascribed predominantly to amyloplasts. Consistently, the analysis of plastid morphology using transmission electron microscopy indicated that starch depletion associated with the proliferation of membrane-free and tubular membrane-containing plastids was a feature specific to arbusculated cells. The loss of enzymes involved in carbon/nitrogen assimilation and provision of reducing power, coupled to macromolecule degradation events in the plastid-enriched fraction of mycorrhizal roots that paralleled lack of starch accumulation in arbusculated cells, lead us to propose that arbuscule functioning elicits a nutrient starvation and an oxidative stress signature that may prime arbuscule breakdown. © 2016 Scandinavian Plant Physiology Society.

Structural changes in plastids of developing Splachnum ampullaceum sporophytes and relationship to odour production

PubMed Central

McCuaig, B.; Dufour, S. C.; Raguso, R. A.; Bhatt, A. P.; Marino, P.

2014-01-01

Many mosses of the family Splachnaceae are entomophilous and rely on flies for spore dispersal. Splachnum ampullaceum produces a yellow- or pink-coloured hypophysis that releases volatile compounds, attracting flies to the mature moss. The biosynthetic sources of the visual and aromatic cues within the hypophysis have not been identified, and may be either symbiotic cyanobacteria or chromoplasts that break down lipids into volatile compounds. Here, we used transmission electron microscopy and gas chromatography-mass spectrometry (GC-MS) to investigate the sources of these attractants, focusing on different tissues and stages of maturation. Microscopy revealed an abundance of plastids within the hypophysis, while no symbiotic bacteria were observed. During plant maturation, plastids differentiated from amyloplasts with large starch granules to photosynthetic chloroplasts and finally to chromoplasts with lipid accumulations. We used GC-MS to identify over 50 volatile organic compounds from mature sporophytes including short-chain oxygenated compounds, unsaturated irregular terpenoids, fatty acid-derived 6- and 8-carbon alcohols and ketones, and the aromatic compounds acetophenone and p-cresol. The hypophysis showed localised production of pungent volatiles, mainly short-chain fermentation compounds and p-cresol. Some of these volatiles have been shown to be produced from lipid oxidase degradation of linolenic acid within chromoplasts. However, other compounds (such as cyclohexanecarboxylic acid esters) may have a microbial origin. Further investigation is necessary to identify the origin of fly attractants in these mosses. PMID:25213550

Post-harvest light treatment increases expression levels of recombinant proteins in transformed plastids of potato tubers.

PubMed

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

2015-09-01

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. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Starch Biosynthesis during Pollen Maturation Is Associated with Altered Patterns of Gene Expression in Maize1

PubMed Central

Datta, Rupali; Chamusco, Karen C.; Chourey, Prem S.

2002-01-01

Starch biosynthesis during pollen maturation is not well understood in terms of genes/proteins and intracellular controls that regulate it in developing pollen. We have studied two specific developmental stages: “early,” characterized by the lack of starch, before or during pollen mitosis I; and “late,” an actively starch-filling post-pollen mitosis I phase in S-type cytoplasmic male-sterile (S-CMS) and two related male-fertile genotypes. The male-fertile starch-positive, but not the CMS starch-deficient, genotypes showed changes in the expression patterns of a large number of genes during this metabolic transition. In addition to a battery of housekeeping genes of carbohydrate metabolism, we observed changes in hexose transporter, plasma membrane H+-ATPase, ZmMADS1, and 14-3-3 proteins. Reduction or deficiency in 14-3-3 protein levels in all three major cellular sites (amyloplasts [starch], mitochondria, and cytosol) in male-sterile relative to male-fertile genotypes are of potential interest because of interorganellar communication in this CMS system. Further, the levels of hexose sugars were significantly reduced in male-sterile as compared with male-fertile tissues, not only at “early” and “late” stages but also at an earlier point during meiosis. Collectively, these data suggest that combined effects of both reduced sugars and their reduced flux in starch biosynthesis along with a strong possibility for altered redox passage may lead to the observed temporal changes in gene expressions, and ultimately pollen sterility. PMID:12481048

Loose Plant Architecture1, an INDETERMINATE DOMAIN Protein Involved in Shoot Gravitropism, Regulates Plant Architecture in Rice1[W

PubMed Central

Wu, Xinru; Tang, Ding; Li, Ming; Wang, Kejian; Cheng, Zhukuan

2013-01-01

Tiller angle and leaf angle are two important components of rice (Oryza sativa) plant architecture that play a crucial role in determining grain yield. Here, we report the cloning and characterization of the Loose Plant Architecture1 (LPA1) gene in rice, the functional ortholog of the AtIDD15/SHOOT GRAVITROPISM5 (SGR5) gene in Arabidopsis (Arabidopsis thaliana). LPA1 regulates tiller angle and leaf angle by controlling the adaxial growth of tiller node and lamina joint. LPA1 was also found to affect shoot gravitropism. Expression pattern analysis suggested that LPA1 influences plant architecture by affecting the gravitropism of leaf sheath pulvinus and lamina joint. However, LPA1 only influences gravity perception or signal transduction in coleoptile gravitropism by regulating the sedimentation rate of amyloplasts, distinct from the actions of LAZY1. LPA1 encodes a plant-specific INDETERMINATE DOMAIN protein and defines a novel subfamily of 28 INDETERMINATE DOMAIN proteins with several unique conserved features. LPA1 is localized in the nucleus and functions as an active transcriptional repressor, an activity mainly conferred by a conserved ethylene response factor-associated amphiphilic repression-like motif. Further analysis suggests that LPA1 participates in a complicated transcriptional and protein interaction network and has evolved novel functions distinct from SGR5. This study not only facilitates the understanding of gravitropism mechanisms but also generates a useful genetic material for rice breeding. PMID:23124325

Auxin, ethylene and light in gravitropic growth: new insights

NASA Astrophysics Data System (ADS)

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

The structure of the fruit peel in two varieties of Malus domestica Borkh. (Rosaceae) before and after storage.

PubMed

Konarska, Agata

2013-06-01

The structure of fruit peel of two apple varieties 'Szampion' and 'Jonagold' was investigated using light microscopy as well as scanning and transmission electron microscopy. The samples were taken immediately after harvest and after 6-month controlled atmosphere storage. The Szampion and Jonagold fruit differed in terms of the surface type, number of lenticels, thickness of the cuticular epithelium, height of epidermal cells and thickness of the hypodermis as well as the amount of crystalline wax and the number of microcracks formed on the fruit surface. The 6-month storage resulted in fruit weight loss, increased numbers and depth of microcracks, thickening of the amorphous wax layer and enhanced production of platelet forms of crystalline wax, which filled the microcracks abundantly. Compared with Jonagold, the Szampion fruit exhibited a fewer lenticels, a bigger number of microcracks, smaller amounts of crystalline wax and more substantial weight loss. The apple varieties studied had a reticulate-lamellate cuticle, and at harvest, the epidermal and hypodermal cells contained numerous amyloplasts filled with starch grains, which were not found after the storage period. Additionally, after storage, the cell protoplasts in the apple peel displayed a disorganised structure, and their vacuoles contained fragments of cell membranes, intravacuolar precipitates and deposits, and spherical bodies. The results may facilitate better understanding of changes occurring in fruits of Szampion and Jonagold during storage and help choose the best storage conditions to reduce loss of weight and prevent impairment of fruit quality.

Nitrogen-fixing nodules induced by Agrobacterium tumefaciens harboring Rhizobium phaseoli plasmids.

PubMed Central

Martínez, E; Palacios, R; Sánchez, F

1987-01-01

Rhizobium phaseoli CFN299 forms nitrogen-fixing nodules in Phaseolus vulgaris (bean) and in Leucaena esculenta. It has three plasmids of 185, 225, and 410 kilobases. The 410-kilobase plasmid contains the nitrogenase structural genes. We have transferred these plasmids to the plasmid-free strain Agrobacterium tumefaciens GMI9023. Transconjugants containing different combinations of the R. phaseoli plasmids were obtained, and they were exhaustively purified before nodulation was assayed. Only transconjugants harboring the 410-kilobase plasmid nodulate P. vulgaris and L. esculenta. Nodules formed by all such transconjugants are able to reduce acetylene. Transconjugants containing the whole set of plasmids from CFN299 nodulate better and fix more nitrogen than the transconjugants carrying only the Sym plasmid. Microscopic analysis of nodules induced by A. tumefaciens transconjugants reveals infected cells and vascular bundles. None of the A. tumefaciens transconjugants, not even the one with the whole set of plasmids from CFN299, behaves in symbiosis like the original R. phaseoli strain; the transconjugants produce fewer nodules and have lower acetylene reduction (25% as compared to the original R. phaseoli strain) and more amyloplasts per nodule. More than 2,000 bacterial isolates from nodules of P. vulgaris and L. esculenta formed by the transconjugants were analyzed by different criteria. Not a single rhizobium could be detected. Our results show that R. phaseoli plasmids may be expressed in the A. tumefaciens background and direct the formation of effective, differentiated nodules. Images PMID:3584072

Starch accumulation in hulless barley during grain filling.

PubMed

Zheng, Xu-Guang; Qi, Jun-Cang; Hui, Hong-Shan; Lin, Li-Hao; Wang, Feng

2017-12-01

Starch consists of two types of molecules: amylose and amylopectin. The objective of this study was increase understanding about mechanisms related to starch accumulation in hulless barley (Hordeum vulgare L.) grain by measuring temporal changes in (i) grain amylose and amylopectin content, (ii) starch synthase activity, and (iii) the relative expressions of key starch-related genes. The amylopectin/amylose ratio gradually declined in both Beiqing 6 and Kunlun 12. In both cultivars, the activities of adenosine diphosphate glucose pyrophosphorylase, soluble starch synthase (SSS), granule bound starch synthase (GBSS), and starch branching enzyme (SBE) increased steadily during grain filling, reaching their maximums 20-25 days after anthesis. The activities of SSS and SBE were greater in Ganken 5 than in either Beiqing 6 or Kunlun 12. The expression of GBSS I was greater in Beiqing 6 and Kunlun 12 than in Ganken 5. In contrast, the expression of SSS I, SSS II and SBE I was greater in Ganken 5 than in Beiqing 6 and Kunlun 12. The peak in GBSS I expression was later than that of SSS I, SSS II, SBE IIa and SBE IIb. The GBSS I transcript in Kunlun 12 was expressed on average 90 times more than the GBSS II transcript. The results suggest that SBE and SSS may control starch synthesis at the transcriptional level, whereas GBSS I may control starch synthesis at the post transcriptional level. GBSS I is mainly responsible for amylose synthesis whereas SSS I and SBE II are mainly responsible for amylopectin synthesis in amyloplasts.

Role of sucrose phosphate synthase in sucrose biosynthesis in ripening bananas and its relationship to the respiratory climacteric.

PubMed

Hubbard, N L; Pharr, D M; Huber, S C

1990-09-01

During ripening of bananas (Musa spp. [AAA group, Cavendish subgroup]), there is a massive conversion of starch to sucrose. Also during ripening there is a rise in respiration known as the respiratory climacteric. In this study changes in carbohydrate content, activities of starch and sucrose metabolizing enzymes, and respiration were measured to assess their potential interrelationships. Sucrose phosphate synthase activity increased dramatically during the first 4 days after initiation of ripening by ethylene treatment. Starch concentration decreased and sucrose concentration increased during this time period. Developmental changes in sucrose phosphate synthase activity were measured with limiting substrate (plus Pi) and saturating substrate concentrations. Activities were not parallel under the two assay conditions, providing tentative evidence that kinetically different forms of the enzyme may exist at different stages of ripening. Sucrose accumulation rate was most highly correlated with sucrose phosphate synthase activity assayed with limiting substrate concentrations (plus Pi). The cumulative amount of CO(2) respired during ripening was positively correlated with sugar accumulation (R(2) = 0.97). From this linear regression it was calculated that a constant 0.605 millimoles of CO(2) was evolved per mole of sucrose formed throughout ripening. Using this quantity, the percentage of the total respiratory ATP produced which was required for the conversion of starch to sucrose was calculated assuming different models for carbon export from the amyloplast. The results suggest that sucrose biosynthesis during ripening constitutes a significant sink for respiratory ATP.

Role of Sucrose Phosphate Synthase in Sucrose Biosynthesis in Ripening Bananas and Its Relationship to the Respiratory Climacteric 1

PubMed Central

Hubbard, Natalie L.; Pharr, D. Mason; Huber, Steven C.

1990-01-01

During ripening of bananas (Musa spp. [AAA group, Cavendish subgroup]), there is a massive conversion of starch to sucrose. Also during ripening there is a rise in respiration known as the respiratory climacteric. In this study changes in carbohydrate content, activities of starch and sucrose metabolizing enzymes, and respiration were measured to assess their potential interrelationships. Sucrose phosphate synthase activity increased dramatically during the first 4 days after initiation of ripening by ethylene treatment. Starch concentration decreased and sucrose concentration increased during this time period. Developmental changes in sucrose phosphate synthase activity were measured with limiting substrate (plus Pi) and saturating substrate concentrations. Activities were not parallel under the two assay conditions, providing tentative evidence that kinetically different forms of the enzyme may exist at different stages of ripening. Sucrose accumulation rate was most highly correlated with sucrose phosphate synthase activity assayed with limiting substrate concentrations (plus Pi). The cumulative amount of CO2 respired during ripening was positively correlated with sugar accumulation (R2 = 0.97). From this linear regression it was calculated that a constant 0.605 millimoles of CO2 was evolved per mole of sucrose formed throughout ripening. Using this quantity, the percentage of the total respiratory ATP produced which was required for the conversion of starch to sucrose was calculated assuming different models for carbon export from the amyloplast. The results suggest that sucrose biosynthesis during ripening constitutes a significant sink for respiratory ATP. PMID:16667688

Loose Plant Architecture1, an INDETERMINATE DOMAIN protein involved in shoot gravitropism, regulates plant architecture in rice.

PubMed

Wu, Xinru; Tang, Ding; Li, Ming; Wang, Kejian; Cheng, Zhukuan

2013-01-01

Tiller angle and leaf angle are two important components of rice (Oryza sativa) plant architecture that play a crucial role in determining grain yield. Here, we report the cloning and characterization of the Loose Plant Architecture1 (LPA1) gene in rice, the functional ortholog of the AtIDD15/SHOOT GRAVITROPISM5 (SGR5) gene in Arabidopsis (Arabidopsis thaliana). LPA1 regulates tiller angle and leaf angle by controlling the adaxial growth of tiller node and lamina joint. LPA1 was also found to affect shoot gravitropism. Expression pattern analysis suggested that LPA1 influences plant architecture by affecting the gravitropism of leaf sheath pulvinus and lamina joint. However, LPA1 only influences gravity perception or signal transduction in coleoptile gravitropism by regulating the sedimentation rate of amyloplasts, distinct from the actions of LAZY1. LPA1 encodes a plant-specific INDETERMINATE DOMAIN protein and defines a novel subfamily of 28 INDETERMINATE DOMAIN proteins with several unique conserved features. LPA1 is localized in the nucleus and functions as an active transcriptional repressor, an activity mainly conferred by a conserved ethylene response factor-associated amphiphilic repression-like motif. Further analysis suggests that LPA1 participates in a complicated transcriptional and protein interaction network and has evolved novel functions distinct from SGR5. This study not only facilitates the understanding of gravitropism mechanisms but also generates a useful genetic material for rice breeding.

Protein-protein interactions among enzymes of starch biosynthesis in high-amylose barley genotypes reveal differential roles of heteromeric enzyme complexes in the synthesis of A and B granules.

PubMed

Ahmed, Zaheer; Tetlow, Ian J; Ahmed, Regina; Morell, Matthew K; Emes, Michael J

2015-04-01

The present study investigated the role of protein phosphorylation, and protein complex formation between key enzymes of amylopectin synthesis, in barley genotypes exhibiting "high amylose" phenotypes. Starch branching enzyme (SBE) down-regulated lines (ΔSBEIIa and ΔSBEIIb), starch synthase (SS)IIa (ssiia(-), sex6) and SSIII (ssiii(-), amo1) mutants were compared to a reference genotype, OAC Baxter. Down-regulation of either SBEIIa or IIb caused pleiotropic effects on SSI and starch phosphorylase (SP) and resulted in formation of novel protein complexes in which the missing SBEII isoform was substituted by SBEI and SP. In the ΔSBEIIb down-regulated line, soluble SP activity was undetectable. Nonetheless, SP was incorporated into a heteromeric protein complex with SBEI and SBEIIa and was readily detected in starch granules. In amo1, unlike other mutants, the data suggest that both SBEIIa and SBEIIb are in a protein complex with SSI and SSIIa. In the sex6 mutant no protein complexes involving SBEIIa or SBEIIb were detected in amyloplasts. Studies with Pro-Q Diamond revealed that GBSS, SSI, SSIIa, SBEIIb and SP are phosphorylated in their granule bound state. Alteration in the granule proteome in ΔSBEIIa and ΔSBEIIb lines, suggests that different protein complexes are involved in the synthesis of A and B granules. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Proteome Profile of Starch Granules Purified from Rice (Oryza sativa) Endosperm.

PubMed

Xing, Shihai; Meng, Xiaoxi; Zhou, Lihui; Mujahid, Hana; Zhao, Chunfang; Zhang, Yadong; Wang, Cailin; Peng, Zhaohua

2016-01-01

Starch is the most important food energy source in cereals. Many of the known enzymes involved in starch biosynthesis are partially or entirely granule-associated in the endosperm. Studying the proteome of rice starch granules is critical for us to further understand the mechanisms underlying starch biosynthesis and packaging of starch granules in rice amyloplasts, consequently for the improvement of rice grain quality. In this article, we developed a protocol to purify starch granules from mature rice endosperm and verified the quality of purified starch granules by microscopy observations, I2 staining, and Western blot analyses. In addition, we found the phenol extraction method was superior to Tris-HCl buffer extraction method with respect to the efficiency in recovery of starch granule associated proteins. LC-MS/MS analysis showed identification of already known starch granule associated proteins with high confidence. Several proteins reported to be involved in starch synthesis in prior genetic studies in plants were also shown to be enriched with starch granules, either directly or indirectly, in our studies. In addition, our results suggested that a few additional candidate proteins may also be involved in starch synthesis. Furthermore, our results indicated that some starch synthesis pathway proteins are subject to protein acetylation modification. GO analysis and KEGG pathway enrichment analysis showed that the identified proteins were mainly located in plastids and involved in carbohydrate metabolism. This study substantially advances the understanding of the starch granule associated proteome in rice and post translational regulation of some starch granule associated proteins.

Molecular genetic analysis of plant gravitropism

NASA Technical Reports Server (NTRS)

Lomax, T. L.

1997-01-01

The analysis of mutants is a powerful approach for elucidating the components of complex biological processes. A growing number of mutants have been isolated which affect plant gravitropism and the classes of mutants found thus far provide important information about the gravity response mechanism. The wide variety of mutants isolated, especially in Arabidopsis, indicates that gravitropism is a complex, multi-step process. The existence of mutants altered in either root gravitropism alone, shoot gravitropism alone, or both indicates that the root and shoot gravitropic mechanisms have both separate and common steps. Reduced starch mutants have confirmed the role of amyloplasts in sensing the gravity signal. The hormone auxin is thought to act as the transducing signal between the sites of gravity perception (the starch parenchyma cells surrounding the vascular tissue in shoots and the columella cells of root caps) and asymmetric growth (the epidermal cells of the elongation zone(s) of each organ). To date, all mutants that are resistant to high concentrations of auxin have also been found to exhibit a reduced gravitropic response, thus supporting the role of auxin. Not all gravitropic mutants are auxin-resistant, however, indicating that there are additional steps which do not involve auxin. Studies with mutants of tomato which exhibit either reduced or reversed gravitropic responses further support the role of auxin redistribution in gravitropism and suggest that both red light and cytokinin interact with gravitropism through controlling lateral auxin transport. Plant responses to gravity thus likely involve changes in both auxin transport and sensitivity.

Columella cells revisited: novel structures, novel properties, and a novel gravisensing model

NASA Technical Reports Server (NTRS)

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

2000-01-01

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.

Transcriptional and Hormonal Regulation of Gravitropism of Woody Stems in Populus.

PubMed

Gerttula, Suzanne; Zinkgraf, Matthew; Muday, Gloria K; Lewis, Daniel R; Ibatullin, Farid M; Brumer, Harry; Hart, Foster; Mansfield, Shawn D; Filkov, Vladimir; Groover, Andrew

2015-10-01

Angiosperm trees reorient their woody stems by asymmetrically producing a specialized xylem tissue, tension wood, which exerts a strong contractile force resulting in negative gravitropism of the stem. Here, we show, in Populus trees, that initial gravity perception and response occurs in specialized cells through sedimentation of starch-filled amyloplasts and relocalization of the auxin transport protein, PIN3. Gibberellic acid treatment stimulates the rate of tension wood formation and gravibending and enhances tissue-specific expression of an auxin-responsive reporter. Gravibending, maturation of contractile fibers, and gibberellic acid (GA) stimulation of tension wood formation are all sensitive to transcript levels of the Class I KNOX homeodomain transcription factor-encoding gene ARBORKNOX2 (ARK2). We generated genome-wide transcriptomes for trees in which gene expression was perturbed by gravistimulation, GA treatment, and modulation of ARK2 expression. These data were employed in computational analyses to model the transcriptional networks underlying wood formation, including identification and dissection of gene coexpression modules associated with wood phenotypes, GA response, and ARK2 binding to genes within modules. We propose a model for gravitropism in the woody stem in which the peripheral location of PIN3-expressing cells relative to the cambium results in auxin transport toward the cambium in the top of the stem, triggering tension wood formation, while transport away from the cambium in the bottom of the stem triggers opposite wood formation. © 2015 American Society of Plant Biologists. All rights reserved.

Structural changes in plastids of developing Splachnum ampullaceum sporophytes and relationship to odour production.

PubMed

McCuaig, B; Dufour, S C; Raguso, R A; Bhatt, A P; Marino, P

2015-03-01

Many mosses of the family Splachnaceae are entomophilous and rely on flies for spore dispersal. Splachnum ampullaceum produces a yellow- or pink-coloured hypophysis that releases volatile compounds, attracting flies to the mature moss. The biosynthetic sources of the visual and aromatic cues within the hypophysis have not been identified, and may be either symbiotic cyanobacteria or chromoplasts that break down lipids into volatile compounds. Here, we used transmission electron microscopy and gas chromatography-mass spectrometry (GC-MS) to investigate the sources of these attractants, focusing on different tissues and stages of maturation. Microscopy revealed an abundance of plastids within the hypophysis, while no symbiotic bacteria were observed. During plant maturation, plastids differentiated from amyloplasts with large starch granules to photosynthetic chloroplasts and finally to chromoplasts with lipid accumulations. We used GC-MS to identify over 50 volatile organic compounds from mature sporophytes including short-chain oxygenated compounds, unsaturated irregular terpenoids, fatty acid-derived 6- and 8-carbon alcohols and ketones, and the aromatic compounds acetophenone and p-cresol. The hypophysis showed localised production of pungent volatiles, mainly short-chain fermentation compounds and p-cresol. Some of these volatiles have been shown to be produced from lipid oxidase degradation of linolenic acid within chromoplasts. However, other compounds (such as cyclohexanecarboxylic acid esters) may have a microbial origin. Further investigation is necessary to identify the origin of fly attractants in these mosses. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

The role of transporters in supplying energy to plant plastids.

PubMed

Flügge, Ulf-Ingo; Häusler, Rainer E; Ludewig, Frank; Gierth, Markus

2011-04-01

The energy status of plant cells strongly depends on the energy metabolism in chloroplasts and mitochondria, which are capable of generating ATP either by photosynthetic or oxidative phosphorylation, respectively. Another energy-rich metabolite inside plastids is the glycolytic intermediate phosphoenolpyruvate (PEP). However, chloroplasts and most non-green plastids lack the ability to generate PEP via a complete glycolytic pathway. Hence, PEP import mediated by the plastidic PEP/phosphate translocator or PEP provided by the plastidic enolase are vital for plant growth and development. In contrast to chloroplasts, metabolism in non-green plastids (amyloplasts) of starch-storing tissues strongly depends on both the import of ATP mediated by the plastidic nucleotide transporter NTT and of carbon (glucose 6-phosphate, Glc6P) mediated by the plastidic Glc6P/phosphate translocator (GPT). Both transporters have been shown to co-limit starch biosynthesis in potato plants. In addition, non-photosynthetic plastids as well as chloroplasts during the night rely on the import of energy in the form of ATP via the NTT. During energy starvation such as prolonged darkness, chloroplasts strongly depend on the supply of ATP which can be provided by lipid respiration, a process involving chloroplasts, peroxisomes, and mitochondria and the transport of intermediates, i.e. fatty acids, ATP, citrate, and oxaloacetate across their membranes. The role of transporters involved in the provision of energy-rich metabolites and in pathways supplying plastids with metabolic energy is summarized here.

ATP-dependent molecular chaperones in plastids--More complex than expected.

PubMed

Trösch, Raphael; Mühlhaus, Timo; Schroda, Michael; Willmund, Felix

2015-09-01

Plastids are a class of essential plant cell organelles comprising photosynthetic chloroplasts of green tissues, starch-storing amyloplasts of roots and tubers or the colorful pigment-storing chromoplasts of petals and fruits. They express a few genes encoded on their organellar genome, called plastome, but import most of their proteins from the cytosol. The import into plastids, the folding of freshly-translated or imported proteins, the degradation or renaturation of denatured and entangled proteins, and the quality-control of newly folded proteins all require the action of molecular chaperones. Members of all four major families of ATP-dependent molecular chaperones (chaperonin/Cpn60, Hsp70, Hsp90 and Hsp100 families) have been identified in plastids from unicellular algae to higher plants. This review aims not only at giving an overview of the most current insights into the general and conserved functions of these plastid chaperones, but also into their specific plastid functions. Given that chloroplasts harbor an extreme environment that cycles between reduced and oxidized states, that has to deal with reactive oxygen species and is highly reactive to environmental and developmental signals, it can be presumed that plastid chaperones have evolved a plethora of specific functions some of which are just about to be discovered. Here, the most urgent questions that remain unsolved are discussed, and guidance for future research on plastid chaperones is given. This article is part of a Special Issue entitled: Chloroplast Biogenesis. Copyright © 2015 Elsevier B.V. All rights reserved.

Development of Gravity Sensitive Plant Cells (Ceratodon) in Microgravity

NASA Technical Reports Server (NTRS)

Sack, Fred D.

1999-01-01

Protonemata of the moss Ceratodon are tip-growing cells that grow up in the dark. This cell type is unique compared to cells in almost any other organism, since the growth of the plant cell itself is completely oriented by gravity. Thus, both the processes of gravity sensing and the gravity response occur in the same cell. Gravity sensing appears to rely upon amyloplasts (starch-filled plastids) that sediment. This sedimentation occurs in specific zones and plastid zonation is complex with respect to plastid morphology, distribution, and gravity. Microtubules restrict the extent of plastid sedimentation (i.e., they are load-bearing). Light also is important since apical cells have a phytochrome-based positive phototropism, light quality influences plastid zonation and sedimentation (photomorphogenesis), and red light suppresses gravitropism at higher but not lower light intensities. Many of these processes were examined in a 16 day spaceflight experiment, "SPM-A" space moss" or "SPAM)) on STS-87 that landed in December, 1997. The work described here involves the definition of a second flight experiment that builds upon the data and questions arising from STS-87. Effort was directed towards further definition of an experiment for the Shuttle (dubbed "SOS" for "Son of SPAM"). Our current target is STS 107 that is scheduled to fly in January 2001. This definition addressed two goals of the STS107 experiment. The goals of the current experiment were to determine whether the cytoskeleton plays a role in maintaining and generating an apical (non-random) plastid distribution in microgravity and to determine the development and extent of clockwise spiral tip-growth in microgravity.

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

PubMed

Nazarian Firouzabadi, Farhad; Kok-Jacon, Géraldine A; Vincken, Jean-Paul; Ji, Qin; Suurs, Luc C J M; Visser, Richard G F

2007-10-01

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 intimate contact with growing starch granules, and to facilitate the incorporation of mutan polymers in starch. Fusion proteins of the appropriate size were evidenced in starch granules, particularly in the amf background. The starches from the various GtfICAT/SBD transformants seemed to contain less mutan than those from transformants with GtfICAT alone, suggesting that the appended SBD might inhibit the activity of GtfICAT in the engineered fusion proteins. Scanning electron microscopy showed that expression of SBD-GtfICAT resulted in alterations of granule morphology in both genetic backgrounds. Surprisingly, the amf starches containing SBD-GtfICAT had a spongeous appearance, i.e., the granule surface contained many small holes and grooves, suggesting that this fusion protein can interfere with the lateral interactions of amylopectin sidechains. No differences in physico-chemical properties of the transgenic starches were observed. Our results show that expression of granule-bound and "soluble" GtfICAT can affect starch biosynthesis differently.

Protein Association and Dissociation Regulated by Ferric Ion

PubMed Central

Li, Chaorui; Fu, Xiaoping; Qi, Xin; Hu, Xiaosong; Chasteen, N. Dennis; Zhao, Guanghua

2009-01-01

Iron stored in phytoferritin plays an important role in the germination and early growth of seedlings. The protein is located in the amyloplast where it stores large amounts of iron as a hydrated ferric oxide mineral core within its shell-like structure. The present work was undertaken to study alternate mechanisms of core formation in pea seed ferritin (PSF). The data reveal a new mechanism for mineral core formation in PSF involving the binding and oxidation of iron at the extension peptide (EP) located on the outer surface of the protein shell. This binding induces aggregation of the protein into large assemblies of ∼400 monomers. The bound iron is gradually translocated to the mineral core during which time the protein dissociates back into its monomeric state. Either the oxidative addition of Fe2+ to the apoprotein to form Fe3+ or the direct addition of Fe3+ to apoPSF causes protein aggregation once the binding capacity of the 24 ferroxidase centers (48 Fe3+/shell) is exceeded. When the EP is enzymatically deleted from PSF, aggregation is not observed, and the rate of iron oxidation is significantly reduced, demonstrating that the EP is a critical structural component for iron binding, oxidation, and protein aggregation. These data point to a functional role for the extension peptide as an iron binding and ferroxidase center that contributes to mineralization of the iron core. As the iron core grows larger, the new pathway becomes less important, and Fe2+ oxidation and deposition occurs directly on the surface of the iron core. PMID:19398557

Touch modulates gravity sensing to regulate the growth of primary roots of Arabidopsis thaliana

NASA Technical Reports Server (NTRS)

Massa, Gioia D.; Gilroy, Simon

2003-01-01

Plants must sense and respond to diverse stimuli to optimize the architecture of their root system for water and nutrient scavenging and anchorage. We have therefore analyzed how information from two of these stimuli, touch and gravity, are integrated to direct root growth. In Arabidopsis thaliana, touch stimulation provided by a glass barrier placed across the direction of growth caused the root to form a step-like growth habit with bends forming in the central and later the distal elongation zones. This response led to the main root axis growing parallel to, but not touching the obstacle, whilst the root cap maintained contact with the barrier. Removal of the graviperceptive columella cells of the root cap using laser ablation reduced the bending response of the distal elongation zone. Similarly, although the roots of the gravisensing impaired pgm1-1 mutant grew along the barrier at the same average angle as wild-type, this angle became more variable with time. These observations imply a constant gravitropic re-setting of the root tip response to touch stimulation from the barrier. In wild-type plants, transient touch stimulation of root cap cells, but not other regions of the root, inhibited both subsequent gravitropic growth and amyloplast sedimentation in the columella. Taken together, these results suggest that the cells of the root cap sense touch stimuli and their subsequent signaling acts on the columella cells to modulate their graviresponse. This interaction of touch and gravity signaling would then direct root growth to avoid obstacles in the soil while generally maintaining downward growth.

The cytoskeleton and gravitropism in higher plants

NASA Technical Reports Server (NTRS)

Blancaflor, Elison B.

2002-01-01

The cellular and molecular mechanisms underlying the gravitropic response of plants have continued to elude plant biologists despite more than a century of research. Lately there has been increased attention on the role of the cytoskeleton in plant gravitropism, but several controversies and major gaps in our understanding of cytoskeletal involvement in gravitropism remain. A major question in the study of plant gravitropism is how the cytoskeleton mediates early sensing and signal transduction events in plants. Much has been made of the actin cytoskeleton as the cellular structure that sedimenting amyloplasts impinge upon to trigger the downstream signaling events leading to the bending response. There is also strong molecular and biochemical evidence that the transport of auxin, an important player in gravitropism, is regulated by actin. Organizational changes in microtubules during the growth response phase of gravitropism have also been well documented, but the significance of such reorientations in controlling differential cellular growth is unclear. Studies employing pharmacological approaches to dissect cytoskeletal involvement in gravitropism have led to conflicting results and therefore need to be interpreted with caution. Despite the current controversies, the revolutionary advances in molecular, biochemical, and cell biological techniques have opened up several possibilities for further research into this difficult area. The myriad proteins associated with the plant cytoskeleton that are being rapidly characterized provide a rich assortment of candidate regulators that could be targets of the gravity signal transduction chain. Cytoskeletal and ion imaging in real time combined with mutant analysis promises to provide a fresh start into this controversial area of research.

Proteome Profile of Starch Granules Purified from Rice (Oryza sativa) Endosperm

PubMed Central

Xing, Shihai; Meng, Xiaoxi; Zhou, Lihui; Mujahid, Hana; Zhao, Chunfang; Zhang, Yadong; Wang, Cailin; Peng, Zhaohua

2016-01-01

Starch is the most important food energy source in cereals. Many of the known enzymes involved in starch biosynthesis are partially or entirely granule-associated in the endosperm. Studying the proteome of rice starch granules is critical for us to further understand the mechanisms underlying starch biosynthesis and packaging of starch granules in rice amyloplasts, consequently for the improvement of rice grain quality. In this article, we developed a protocol to purify starch granules from mature rice endosperm and verified the quality of purified starch granules by microscopy observations, I2 staining, and Western blot analyses. In addition, we found the phenol extraction method was superior to Tris-HCl buffer extraction method with respect to the efficiency in recovery of starch granule associated proteins. LC-MS/MS analysis showed identification of already known starch granule associated proteins with high confidence. Several proteins reported to be involved in starch synthesis in prior genetic studies in plants were also shown to be enriched with starch granules, either directly or indirectly, in our studies. In addition, our results suggested that a few additional candidate proteins may also be involved in starch synthesis. Furthermore, our results indicated that some starch synthesis pathway proteins are subject to protein acetylation modification. GO analysis and KEGG pathway enrichment analysis showed that the identified proteins were mainly located in plastids and involved in carbohydrate metabolism. This study substantially advances the understanding of the starch granule associated proteome in rice and post translational regulation of some starch granule associated proteins. PMID:27992503

High day- and night-time temperatures affect grain growth dynamics in contrasting rice genotypes.

PubMed

Shi, Wanju; Yin, Xinyou; Struik, Paul C; Solis, Celymar; Xie, Fangming; Schmidt, Ralf C; Huang, Min; Zou, Yingbin; Ye, Changrong; Jagadish, S V Krishna

2017-11-02

Rice grain yield and quality are predicted to be highly vulnerable to global warming. Five genotypes including heat-tolerant and susceptible checks, a heat-tolerant near-isogenic line and two hybrids were exposed to control (31 °C/23 °C, day/night), high night-time temperature (HNT; 31 °C/30 °C), high day-time temperature (HDT; 38 °C/23 °C) and high day- and night-time temperature (HNDT; 38 °C/30 °C) treatments for 20 consecutive days during the grain-filling stage. Grain-filling dynamics, starch metabolism enzymes, temporal starch accumulation patterns and the process of chalk formation were quantified. Compensation between the rate and duration of grain filling minimized the impact of HNT, but irreversible impacts on seed-set, grain filling and ultimately grain weight were recorded with HDT and HNDT. Scanning electron microscopy demonstrated irregular and smaller starch granule formation affecting amyloplast build-up with HDT and HNDT, while a quicker but normal amylopast build-up was recorded with HNT. Our findings revealed temporal variation in the starch metabolism enzymes in all three stress treatments. Changes in the enzymatic activity did not derail starch accumulation under HNT when assimilates were sufficiently available, while both sucrose supply and the conversion of sucrose into starch were affected by HDT and HNDT. The findings indicate differential mechanisms leading to high day and high night temperature stress-induced loss in yield and quality. Additional genetic improvement is needed to sustain rice productivity and quality under future climates. © Society for Experimental Biology 2017.

Root gravitropism: an experimental tool to investigate basic cellular and molecular processes underlying mechanosensing and signal transmission in plants

NASA Technical Reports Server (NTRS)

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

2002-01-01

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.

Calcium movement, graviresponsiveness and the structure of columella cells and columella tissues in roots of Allium cepa L

NASA Technical Reports Server (NTRS)

Moore, R.

1985-01-01

Roots of Allium cepa L. cv. Yellow are differentially responsive to gravity. Long (e.g. 40 mm) roots are strongly graviresponsive, while short (c.g. 4 mm) roots are minimally responsive to gravity. Although columella cells of graviresponsive roots are larger than those of nongraviresponsive roots, they partition their volumes to cellular organelles similarly. The movement of amyloplasts and nuclei in columella cells of horizontally-oriented roots correlates positively with the onset of gravicurvature. Furthermore, there is no significant difference in the rates of organellar redistribution when graviresponsive and nongraviresponsive roots are oriented horizontally. The more pronounced graviresponsiveness of longer roots correlates positively with (1) their caps being 9-6 times more voluminous, (2) their columella tissues being 42 times more voluminous, (3) their caps having 15 times more columella cells, and (4) their columella tissues having relative volumes 4.4 times larger than those of shorter, nongraviresponsive roots. Graviresponsive roots that are oriented horizontally are characterized by a strongly polar movement of 45Ca2+ across the root tip from the upper to the lower side, while similarly oriented nongraviresponsive roots exhibit only a minimal polar transport of 45Ca2+. These results indicate that the differential graviresponsiveness of roots of A. cepa is probably not due to either (1) ultrastructural differences in their columella cells, (2) differences in the rates of organellar redistribution when roots are oriented horizontally. Rather, these results indicate the graviresponsiveness may require an extensive columella tissue, which, in turn, may be necessary for polar movement of 45Ca2+ across the root tip.

Reduced gravitropic sensitivity in roots of a starch-deficient mutant of Nicotiana sylvestris

NASA Technical Reports Server (NTRS)

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

1989-01-01

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 2.7.5.1.). 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 < or = 5 s for WT roots and 30-60 s for mutant roots. The growth rates for WT and NS 458 roots were essentially equal. These results and our previous results with WT and starchless mutant Arabidopsis roots (Kiss et al. 1989, 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.

Ionic signaling in plant responses to gravity and touch

NASA Technical Reports Server (NTRS)

Fasano, Jeremiah M.; Massa, Gioia D.; Gilroy, Simon

2002-01-01

Touch and gravity are two of the many stimuli that plants must integrate to generate an appropriate growth response. Due to the mechanical nature of both of these signals, shared signal transduction elements could well form the basis of the cross-talk between these two sensory systems. However, touch stimulation must elicit signaling events across the plasma membrane whereas gravity sensing is thought to represent transformation of an internal force, amyloplast sedimentation, to signal transduction events. In addition, factors such as turgor pressure and presence of the cell wall may also place unique constraints on these plant mechanosensory systems. Even so, the candidate signal transduction elements in both plant touch and gravity sensing, changes in Ca2+, pH and membrane potential, do mirror the known ionic basis of signaling in animal mechanosensory cells. Distinct spatial and temporal signatures of Ca2+ ions may encode information about the different mechanosignaling stimuli. Signals such as Ca2+ waves or action potentials may also rapidly transfer information perceived in one cell throughout a tissue or organ leading to the systemic reactions characteristic of plant touch and gravity responses. Longer-term growth responses are likely sustained via changes in gene expression and asymmetries in compounds such as inositol-1,4,5-triphosphate (IP3) and calmodulin. Thus, it seems likely that plant mechanoperception involves both spatial and temporal encoding of information at all levels, from the cell to the whole plant. Defining this patterning will be a critical step towards understanding how plants integrate information from multiple mechanical stimuli to an appropriate growth response.

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

NASA Technical Reports Server (NTRS)

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

1999-01-01

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.

Comparative ultrastructure of fruit plastids in three genetically diverse genotypes of apple (Malus × domestica Borkh.) during development

PubMed Central

Schaeffer, Scott M.; Christian, Ryan; Castro-Velasquez, Nohely; Hyden, Brennan; Lynch-Holm, Valerie

2017-01-01

Plastids are the defining organelle for a plant cell and are critical for myriad metabolic functions. The role of leaf plastid, chloroplast, is extensively documented; however, fruit plastids—chromoplasts—are poorly understood, especially in the context of the diverse metabolic processes operating in these diverse plant organs. Recently, in a comparative study of the predicted plastid-targeted proteomes across seven plant species, we reported that each plant species is predicted to harbor a unique set of plastid-targeted proteins. However, the temporal and developmental context of these processes remains unknown. In this study, an ultrastructural analysis approach was used to characterize fruit plastids in the epidermal and collenchymal cell layers at 11 developmental timepoints in three genotypes of apple (Malus × domestica Borkh.): chlorophyll-predominant ‘Granny Smith’, carotenoid-predominant ‘Golden Delicious’, and anthocyanin-predominant ‘Top Red Delicious’. Plastids transitioned from a proplastid-like plastid to a chromoplast-like plastid in epidermis cells, while in the collenchyma cells, they transitioned from a chloroplast-like plastid to a chloro-chromo-amyloplast plastid. Plastids in the collenchyma cells of the three genotypes demonstrated a diverse array of structures and features. This study enabled the identification of discrete developmental stages during which specific functions are most likely being performed by the plastids as indicated by accumulation of plastoglobuli, starch granules, and other sub-organeller structures. Information regarding the metabolically active developmental stages is expected to facilitate biologically relevant omics studies to unravel the complex biochemistry of plastids in perennial non-model systems. PMID:28698906

Regulatory Shifts in Plastid Transcription Play a Key Role in Morphological Conversions of Plastids during Plant Development.

PubMed

Liebers, Monique; Grübler, Björn; Chevalier, Fabien; Lerbs-Mache, Silva; Merendino, Livia; Blanvillain, Robert; Pfannschmidt, Thomas

2017-01-01

Plastids display a high morphological and functional diversity. Starting from an undifferentiated small proplastid, these plant cell organelles can develop into four major forms: etioplasts in the dark, chloroplasts in green tissues, chromoplasts in colored flowers and fruits and amyloplasts in roots. The various forms are interconvertible into each other depending on tissue context and respective environmental condition. Research of the last two decades uncovered that each plastid type contains its own specific proteome that can be highly different from that of the other types. Composition of these proteomes largely defines the enzymatic functionality of the respective plastid. The vast majority of plastid proteins is encoded in the nucleus and must be imported from the cytosol. However, a subset of proteins of the photosynthetic and gene expression machineries are encoded on the plastid genome and are transcribed by a complex transcriptional apparatus consisting of phage-type nuclear-encoded RNA polymerases and a bacterial-type plastid-encoded RNA polymerase. Both types recognize specific sets of promoters and transcribe partly over-lapping as well as specific sets of genes. Here we summarize the current knowledge about the sequential activity of these plastid RNA polymerases and their relative activities in different types of plastids. Based on published plastid gene expression profiles we hypothesize that each conversion from one plastid type into another is either accompanied or even preceded by significant changes in plastid transcription suggesting that these changes represent important determinants of plastid morphology and protein composition and, hence, the plastid type.

Chromoplasts ultrastructure and estimated carotene content in root secondary phloem of different carrot varieties.

PubMed

Kim, Ji Eun; Rensing, Kim H; Douglas, Carl J; Cheng, Kimberly M

2010-02-01

There have been few studies on quantifying carotenoid accumulation in carrots, and none have taken the comparative approach. The abundance and distribution of carotenes in carrot roots of three varieties, white, orange, and high carotene mass (HCM) were compared using light and transmission electron microscopy (TEM). Light microscopy has indicated that, in all three varieties, carotenes were most abundant in the secondary phloem and this area was selected for further TEM analysis. While carotenes were extracted during the fixation process for TEM, the high-pressure freezing technique we employed preserved the spaces (CS) left behind by the extracted carotene crystals. Chromoplasts from the HCM variety contained significantly (P < 0.05) more CS than chromoplasts from the orange variety. Chromoplasts from the white variety had few or no CS. There was no significant difference between the HCM and orange varieties in the number of chromoplasts per unit area, but the white variety had significantly (P < 0.05) fewer chromoplasts than the other two varieties. A large number of starch-filled amyloplasts was observed in secondary phloem of the white variety but these were not found in the other two varieties. The results from this comparative approach clearly define the subcellular localization of carotenoids in carrot roots and suggest that while the HCM genotype was selectively bred for increased carotene content, this selection did not lead to increased numbers of carotene-containing chromoplasts but rather greater accumulation of carotene per chromoplast. Furthermore, the results confirm that roots of the white carrot variety retain residual amounts of carotene.

Regulatory Shifts in Plastid Transcription Play a Key Role in Morphological Conversions of Plastids during Plant Development

PubMed Central

Liebers, Monique; Grübler, Björn; Chevalier, Fabien; Lerbs-Mache, Silva; Merendino, Livia; Blanvillain, Robert; Pfannschmidt, Thomas

2017-01-01

Plastids display a high morphological and functional diversity. Starting from an undifferentiated small proplastid, these plant cell organelles can develop into four major forms: etioplasts in the dark, chloroplasts in green tissues, chromoplasts in colored flowers and fruits and amyloplasts in roots. The various forms are interconvertible into each other depending on tissue context and respective environmental condition. Research of the last two decades uncovered that each plastid type contains its own specific proteome that can be highly different from that of the other types. Composition of these proteomes largely defines the enzymatic functionality of the respective plastid. The vast majority of plastid proteins is encoded in the nucleus and must be imported from the cytosol. However, a subset of proteins of the photosynthetic and gene expression machineries are encoded on the plastid genome and are transcribed by a complex transcriptional apparatus consisting of phage-type nuclear-encoded RNA polymerases and a bacterial-type plastid-encoded RNA polymerase. Both types recognize specific sets of promoters and transcribe partly over-lapping as well as specific sets of genes. Here we summarize the current knowledge about the sequential activity of these plastid RNA polymerases and their relative activities in different types of plastids. Based on published plastid gene expression profiles we hypothesize that each conversion from one plastid type into another is either accompanied or even preceded by significant changes in plastid transcription suggesting that these changes represent important determinants of plastid morphology and protein composition and, hence, the plastid type. PMID:28154576

Effects of nickel on the chloroplasts of the duckweeds Spirodela polyrhiza and Lemna minor and their possible use in biomonitoring and phytoremediation.

PubMed

Appenroth, K-J; Krech, K; Keresztes, A; Fischer, W; Koloczek, H

2010-01-01

Toxic effects of Ni(2+) on the chloroplasts of the two duckweed species Spirodela polyrhiza, clone SJ and Lemna minor, clone St were investigated according to the ISO 20079 protocol. Ni(2+) induced a transition from chloroplasts to chloro-amyloplasts and amylo-chloroplasts, but not to gerontoplasts, as shown by electron microscopy. The contents of the chlorophylls a and b decreased strongly, whereas that of carotenoids remained approximately constant. Most striking was, however, the accumulation of transitory starch. Bell-shaped dose-response curves showed that Spirodela and Lemna amassed maximum starch contents of approximately 10% and 7%, respectively, on a fresh weight basis. Because Ni(2+) in the concentrations applied does not stimulate photosynthesis, the Ni(2+)-induced starch accumulation indicates that the export of carbohydrates out of the plastids decreased, most probably due to the lower demand of the rest of the cells as a result of the Ni(2+)-dependent inhibition of growth. The half-maximal concentrations for inhibition of the fresh weight increase over the 7-day test period were 3.7 microM and 6.6 microM for Spirodela and Lemna, respectively: Spirodela was thus somewhat more sensitive to the heavy metal. Both species accumulated approximately 3g of Ni(2+) per kg of dry weight after application of 100 microM NiCl(2). Because of their high sensitivity to phytotoxic effects, however, Spirodela and Lemna do not appear to be particularly suitable for phytoremediation of Ni(2+)-contaminated waste water. The high sensitivity to Ni(2+) makes them instead a suitable system for ecotoxicological testing in accordance with the ISO 20079 protocol. Copyright 2009 Elsevier Ltd. All rights reserved.

Mechanosensitive channels are activated by stress in the actin stress fibres, and could be involved in gravity sensing in plants.

PubMed

Tatsumi, H; Furuichi, T; Nakano, M; Toyota, M; Hayakawa, K; Sokabe, M; Iida, H

2014-01-01

Mechanosensitive (MS) channels are expressed in a variety of cells. The molecular and biophysical mechanism involved in the regulation of MS channel activities is a central interest in basic biology. MS channels are thought to play crucial roles in gravity sensing in plant cells. To date, two mechanisms have been proposed for MS channel activation. One is that tension development in the lipid bilayer directly activates MS channels. The second mechanism proposes that the cytoskeleton is involved in the channel activation, because MS channel activities are modulated by pharmacological treatments that affect the cytoskeleton. We tested whether tension in the cytoskeleton activates MS channels. Mammalian endothelial cells were microinjected with phalloidin-conjugated beads, which bound to stress fibres, and a traction force to the actin cytoskeleton was applied by dragging the beads with optical tweezers. MS channels were activated when the force was applied, demonstrating that a sub-pN force to the actin filaments activates a single MS channel. Plants may use a similar molecular mechanism in gravity sensing, since the cytoplasmic Ca(2+) concentration increase induced by changes in the gravity vector was attenuated by potential MS channel inhibitors, and by actin-disrupting drugs. These results support the idea that the tension increase in actin filaments by gravity-dependent sedimentation of amyloplasts activates MS Ca(2+) -permeable channels, which can be the molecular mechanism of a Ca(2+) concentration increase through gravistimulation. We review recent progress in the study of tension sensing by actin filaments and MS channels using advanced biophysical methods, and discuss their possible roles in gravisensing. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.

Gravity resistance, another graviresponse in plants - role of microtubule-membrane-cell wall continuum

NASA Astrophysics Data System (ADS)

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.

Integration of orientation, clinorotation, and sensitivity in the graviresponse

NASA Astrophysics Data System (ADS)

Hasenstein, Karl H.; John, Susan

Gravitropism describes the response of plants to some acceleration and typically involves amy-loplast displacement. Despite numerous studies, opinions diverge on the perception threshold and persistence of the stimulus. Short-term reorientation in the gravity field and superim-posed mechanostimulation (clinorotation) has the potential to reveal the duration (half-life) of mechanostimulation or memory persistence. Although clinorotation is commonly used to compensate the effect of continuous gravity stimulation, it adds long-term mechanostimulation to short-term reorientation. The constantly changing gravity vector is likely to interfere with curvature and root development. Especially sensitive are columella cells and entire layers have been shown to undergo programmed cell death upon clinorotation. It is unknown to what extent this response depends on the onset, speed, or duration of clinorotation. To determine the susceptibility of the graviresponse on clinorotation, we studied the effects of the rate (0.5 to 5 rpm) of rotation of reoriented flax roots (placed horizontally for 5, 10, or 15 min). Seedlings were kept in the dark, clinorotated either parallel or perpendicular to the root axis and imaged after each rotation by an infrared video camera. Horizontal clinorotation did not affect root growth rate (0.78±0.05 mm/h) but vertical clinorotation reduced root growth by more than 10%. The rate of clinorotation did not affect growth for either condition. However, maximal curvature for vertical clinorotation decreased with increasing rate of rotation and produced straight roots at 5 rpm. Horizontal clinorotation increased curvature with increasing reorienta-tion time and resulted in curvature that increased with the rotation rate. Thus, clinorotation changes the graviresponse but introduces secondary effects that depend on rate and direction of rotation.

A Bird's-Eye View of Molecular Changes in Plant Gravitropism Using Omics Techniques.

PubMed

Schüler, Oliver; Hemmersbach, Ruth; Böhmer, Maik

2015-01-01

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 gravitational stimulus are less well understood. Amyloplasts, localized in statocytes of the root tip, in mesophyll 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. Downstream signaling events, including the conversion from the biophysical signal of sedimentation 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 identi-fication 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 review aims at summarizing recent experimental approaches and discusses major observations.

The glutathione peroxidase gene family of Lotus japonicus: characterization of genomic clones, expression analyses and immunolocalization in legumes.

PubMed

Ramos, Javier; Matamoros, Manuel A; Naya, Loreto; James, Euan K; Rouhier, Nicolas; Sato, Shusei; Tabata, Satoshi; Becana, Manuel

2009-01-01

Despite the multiple roles played by antioxidants in rhizobia-legume symbioses, little is known about glutathione peroxidases (GPXs) in legumes. Here the characterization of six GPX genes of Lotus japonicus is reported. Expression of GPX genes was analysed by quantitative reverse transcription-polymerase chain reaction in L. japonicus and Lotus corniculatus plants exposed to various treatments known to generate reactive oxygen and/or nitrogen species. LjGPX1 and LjGPX3 were the most abundantly expressed genes in leaves, roots and nodules. Compared with roots, LjGPX1 and LjGPX6 were highly expressed in leaves and LjGPX3 and LjGPX6 in nodules. In roots, salinity decreased GPX4 expression, aluminium decreased expression of the six genes, and cadmium caused up-regulation of GPX3, GPX4 and GPX5 after 1 h and down-regulation of GPX1, GPX2, GPX4 and GPX6 after 3-24 h. Exposure of roots to sodium nitroprusside (a nitric oxide donor) for 1 h increased the mRNA levels of GPX4 and GPX6 by 3.3- and 30-fold, respectively. Thereafter, the GPX6 mRNA level remained consistently higher than that of the control. Immunogold labelling revealed the presence of GPX proteins in root and nodule amyloplasts and in leaf chloroplasts of L. japonicus and other legumes. Labelling was associated with starch grains. These results underscore the differential regulation of GPX expression in response to cadmium, aluminium and nitric oxide, and strongly support a role for GPX6 and possibly other GPX genes in stress and/or metabolic signalling.

Characterization of root agravitropism induced by genetic, chemical, and developmental constraints

NASA Technical Reports Server (NTRS)

Moore, R.; Fondren, W. M.; Marcum, H.

1987-01-01

The patterns and rates of organelle redistribution in columella (i.e., putative statocyte) cells of agravitropic agt mutants of Zea mays are not significantly different from those of columella cells in graviresponsive roots. Graviresponsive roots of Z. mays are characterized by a strongly polar movement of 45Ca2+ across the root tip from the upper to the lower side. Horizontally-oriented roots of agt mutants exhibit only a minimal polar transport of 45Ca2+. Exogenously-induced asymmetries of Ca result in curvature of agt roots toward the Ca source. A similar curvature can be induced by a Ca asymmetry in normally nongraviresponsive (i.e., lateral) roots of Phaseolus vulgaris. Similarly, root curvature can be induced by placing the roots perpendicular to an electric field. This electrotropism increased with 1) currents between 8-35 mA, and 2) time between 1-9 hr when the current is constant. Electrotropism is reduced significantly by treating roots with triiodobenzoic acid (TIBA), an inhibitor of auxin transport. These results suggest that 1) if graviperception occurs via the sedimentation of amyloplasts in columella cells, then nongraviresponsive roots apparently sense gravity as do graviresponsive roots, 2) exogenously-induced asymmetries of a gravitropic effector (i.e., Ca) can induce curvature of normally nongraviresponsive roots, 3) the gravity-induced downward movement of exogenously-applied 45Ca2+ across tips of graviresponsive roots does not occur in nongraviresponsive roots, 4) placing roots in an electrical field (i.e., one favoring the movement of ions such as Ca2+) induces root curvature, and 5) electrically-induced curvature is apparently dependent on auxin transport. These results are discussed relative to a model to account for the lack of graviresponsiveness by these roots.

A Bird’s-Eye View of Molecular Changes in Plant Gravitropism Using Omics Techniques

PubMed Central

Schüler, Oliver; Hemmersbach, Ruth; Böhmer, Maik

2015-01-01

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 gravitational stimulus are less well understood. Amyloplasts, localized in statocytes of the root tip, in mesophyll 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. Downstream signaling events, including the conversion from the biophysical signal of sedimentation 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 identi-fication 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 review aims at summarizing recent experimental approaches and discusses major observations. PMID:26734055

Comparative ultrastructure of fruit plastids in three genetically diverse genotypes of apple (Malus × domestica Borkh.) during development.

PubMed

Schaeffer, Scott M; Christian, Ryan; Castro-Velasquez, Nohely; Hyden, Brennan; Lynch-Holm, Valerie; Dhingra, Amit

2017-10-01

Comparative ultrastructural developmental time-course analysis has identified discrete stages at which the fruit plastids undergo structural and consequently functional transitions to facilitate subsequent development-guided understanding of the complex plastid biology. Plastids are the defining organelle for a plant cell and are critical for myriad metabolic functions. The role of leaf plastid, chloroplast, is extensively documented; however, fruit plastids-chromoplasts-are poorly understood, especially in the context of the diverse metabolic processes operating in these diverse plant organs. Recently, in a comparative study of the predicted plastid-targeted proteomes across seven plant species, we reported that each plant species is predicted to harbor a unique set of plastid-targeted proteins. However, the temporal and developmental context of these processes remains unknown. In this study, an ultrastructural analysis approach was used to characterize fruit plastids in the epidermal and collenchymal cell layers at 11 developmental timepoints in three genotypes of apple (Malus × domestica Borkh.): chlorophyll-predominant 'Granny Smith', carotenoid-predominant 'Golden Delicious', and anthocyanin-predominant 'Top Red Delicious'. Plastids transitioned from a proplastid-like plastid to a chromoplast-like plastid in epidermis cells, while in the collenchyma cells, they transitioned from a chloroplast-like plastid to a chloro-chromo-amyloplast plastid. Plastids in the collenchyma cells of the three genotypes demonstrated a diverse array of structures and features. This study enabled the identification of discrete developmental stages during which specific functions are most likely being performed by the plastids as indicated by accumulation of plastoglobuli, starch granules, and other sub-organeller structures. Information regarding the metabolically active developmental stages is expected to facilitate biologically relevant omics studies to unravel the complex biochemistry of plastids in perennial non-model systems.

Pseudopollen in Eria Lindl. section Mycaranthes Rchb.f. (Orchidaceae).

PubMed

Davies, K L; Turner, M P

2004-11-01

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. 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. 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 other continents.

Peroxidases, lignin and anatomy during in vitro and ex vitro rooting of gardenia (Gardenia jasminoides Ellis) microshoots.

PubMed

Hatzilazarou, Stefanos P; Syros, Thomas D; Yupsanis, Traianos A; Bosabalidis, Artemios M; Economou, Athanasios S

2006-07-01

In vitro and ex vitro rooting of gardenia (Gardenia jasminoides Ellis) microshoots with or without indolic-3-butyric acid (IBA) was studied in order to improve acclimatization of microplants after root formation and transplantation. Peroxidase (POD) activity and isoforms, lignin content and anatomical observations were evaluated in the course of the three interdependent phases (induction, initiation and expression) of microshoot rooting. Microshoots treated or not treated with IBA achieved high rooting percentages both in vitro and ex vitro. At the end of the 2-week acclimatization period, the percentage of surviving microplants ranged from 80% to 100%, for in vitro and ex vitro rooted microshoots, respectively. Microshoots rooted in vitro and ex vitro showed a relationship between rooting and POD activity but in a different time course. It appeared that root formation occurred after the microshoots had reached and passed a peak of maximum enzyme activity. In all treatments, electrophoretic analysis (native PAGE) of PODs revealed the appearance of one anionic and three cationic POD isoforms (C(1), C(3) and C(4)). An additional cationic POD isoform (C(2)) appeared only in the ex vitro rooting. The lignin content was similar in microshoots rooted both in vitro and ex vitro. The sequential anatomical changes during the rooting process were similar in both in vitro and ex vitro rooting treatments. In the case of in vitro rooting, pith cells had vacuoles entirely filled with a dark substance, while in the case of ex vitro rooting, pith cells contained many amyloplasts. The origin of the adventitious roots, in both rooting conditions, was located in the cambial ring. Roots with organized tissue systems emerged from the microshoot stem 10-14 days after the root induction treatments; on day 10 for rooting in vitro, while a 4-day delay was noted in microshoots rooted ex vitro.

Identification and analysis of novel genes involved in gravitropism of Arabidopsis thaliana.

NASA Astrophysics Data System (ADS)

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

2012-07-01

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.

Isolation and Characterization of Ftsz Genes in Cassava.

PubMed

Geng, Meng-Ting; Min, Yi; Yao, Yuan; Chen, Xia; Fan, Jie; Yuan, Shuai; Wang, Lei; Sun, Chong; Zhang, Fan; Shang, Lu; Wang, Yun-Lin; Li, Rui-Mei; Fu, Shao-Ping; Duan, Rui-Jun; Liu, Jiao; Hu, Xin-Wen; Guo, Jian-Chun

2017-12-15

The filamenting temperature-sensitive Z proteins (FtsZs) play an important role in plastid division. In this study, three FtsZ genes were isolated from the cassava genome, and named MeFtsZ1, MeFtsZ2-1, and MeFtsZ2-2, respectively. Based on phylogeny, the MeFtsZs were classified into two groups (FtsZ1 and FtsZ2). MeFtsZ1 with a putative signal peptide at N-terminal, has six exons, and is classed to FtsZ1 clade. MeFtsZ2-1 and MeFtsZ2-2 without a putative signal peptide, have seven exons, and are classed to FtsZ2 clade. Subcellular localization found that all the three MeFtsZs could locate in chloroplasts and form a ring in chloroplastids. Structure analysis found that all MeFtsZ proteins contain a conserved guanosine triphosphatase (GTPase) domain in favor of generate contractile force for cassava plastid division. The expression profiles of MeFtsZ genes by quantitative reverse transcription-PCR (qRT-PCR) analysis in photosynthetic and non-photosynthetic tissues found that all of the MeFtsZ genes had higher expression levels in photosynthetic tissues, especially in younger leaves, and lower expression levels in the non-photosynthetic tissues. During cassava storage root development, the expressions of MeFtsZ2-1 and MeFtsZ2-2 were comparatively higher than MeFtsZ1. The transformed Arabidopsis of MeFtsZ2-1 and MeFtsZ2-2 contained abnormally shape, fewer number, and larger volume chloroplasts. Phytohormones were involved in regulating the expressions of MeFtsZ genes. Therefore, we deduced that all of the MeFtsZs play an important role in chloroplast division, and that MeFtsZ 2 (2-1, 2-2) might be involved in amyloplast division and regulated by phytohormones during cassava storage root development.

Purification of a NAD(P) reductase-like protein from the thermogenic appendix of the Sauromatum guttatum inflorescence.

PubMed

Skubatz, Hanna; Howald, William N

2013-03-01

A NAD(P) reductase-like protein with a molecular mass of 34.146 ± 34 Da was purified to homogeneity from the appendix of the inflorescence of the Sauromatum guttatum. On-line liquid chromatography/electrospray ionization-mass spectrometry was used to isolate and quantify the protein. For the identification of the protein, liquid chromatography/electrospray ionization-tandem mass spectrometry analysis of tryptic digests of the protein was carried out. The acquired mass spectra were used for database searching, which led to the identification of a single tryptic peptide. The 12 amino acid tryptic peptide (FLPSEFGNDVDR) was found to be identical to amino acid residues at the positions 108-120 of isoflavone reductase in the Arabidopsis genome. A BLAST search identified this sequence region as unique and specific to a class of NAD(P)-dependent reductases involved in phenylpropanoid biosynthesis. Edman degradation revealed that the protein was N-terminally blocked. The amount of the protein (termed RL, NAD(P) reductase-like protein) increased 60-fold from D-4 (4 days before inflorescence-opening, designated as D-day) to D-Day, and declined the following day, when heat-production ceased. When salicylic acid, the endogenous trigger of heat-production in the Sauromatum appendix, was applied to premature appendices, a fivefold decrease in the amount of RL was detected in the treated section relative to the non-treated section. About 40 % of RL was found in the cytoplasm. Another 30 % was detected in Percoll-purified mitochondria and the rest, about 30 % was associated with a low speed centrifugation pellet due to nuclei and amyloplast localization. RL was also found in other thermogenic plants and detected in Arabidopsis leaves. The function of RL in thermogenic and non-thermogenic plants requires further investigation.

New insights into Fe localization in plant tissues

PubMed Central

Roschzttardtz, Hannetz; Conéjéro, Geneviève; Divol, Fanchon; Alcon, Carine; Verdeil, Jean-Luc; Curie, Catherine; Mari, Stéphane

2013-01-01

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. PMID:24046774

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

PubMed Central

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

2004-01-01

• 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 other continents. PMID:15451721

Amyloplast sedimentation dynamics in maize columella cells support a new model for the gravity-sensing apparatus of roots

NASA Technical Reports Server (NTRS)

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

2001-01-01

Quantitative analysis of statolith sedimentation behavior was accomplished using videomicroscopy of living columella cells of corn (Zea mays) roots, which displayed no systematic cytoplasmic streaming. Following 90 degrees rotation of the root, the statoliths moved downward along the distal wall and then spread out along the bottom with an average velocity of 1.7 microm min(-1). When statolith trajectories traversed the complete width or length of the cell, they initially moved horizontally toward channel-initiation sites and then moved vertically through the channels to the lower side of the reoriented cell where they again dispersed. These statoliths exhibited a significantly lower average velocity than those sedimenting on distal-to-side trajectories. In addition, although statoliths undergoing distal-to-side sedimentation began at their highest velocity and slowed monotonically as they approached the lower cell membrane, statoliths crossing the cell's central region remained slow initially and accelerated to maximum speed once they reached a channel. The statoliths accelerated sooner, and the channeling effect was less pronounced in roots treated with cytochalasin D. Parallel ultrastructural studies of high-pressure frozen-freeze-substituted columella cells suggest that the low-resistance statolith pathway in the cell periphery corresponds to the sharp interface between the endoplasmic reticulum (ER)-rich cortical and the ER-devoid central region of these cells. The central region is also shown to contain an actin-based cytoskeletal network in which the individual, straight, actin-like filaments are randomly distributed. To explain these findings as well as the results of physical simulation experiments, we have formulated a new, tensegrity-based model of gravity sensing in columella cells. This model envisages the cytoplasm as pervaded by an actin-based cytoskeletal network that is denser in the ER-devoid central region than in the ER-rich cell cortex and is linked to stretch receptors in the plasma membrane. Sedimenting statoliths are postulated to produce a directional signal by locally disrupting the network and thereby altering the balance of forces acting on the receptors in different plasma membrane regions.

Amyloplast Sedimentation Dynamics in Maize Columella Cells Support a New Model for the Gravity-Sensing Apparatus of Roots1

PubMed Central

Yoder, Thomas L.; Zheng, Hui-qiong; Todd, Paul; Staehelin, L. Andrew

2001-01-01

Quantitative analysis of statolith sedimentation behavior was accomplished using videomicroscopy of living columella cells of corn (Zea mays) roots, which displayed no systematic cytoplasmic streaming. Following 90° rotation of the root, the statoliths moved downward along the distal wall and then spread out along the bottom with an average velocity of 1.7 μm min−1. When statolith trajectories traversed the complete width or length of the cell, they initially moved horizontally toward channel-initiation sites and then moved vertically through the channels to the lower side of the reoriented cell where they again dispersed. These statoliths exhibited a significantly lower average velocity than those sedimenting on distal-to-side trajectories. In addition, although statoliths undergoing distal-to-side sedimentation began at their highest velocity and slowed monotonically as they approached the lower cell membrane, statoliths crossing the cell's central region remained slow initially and accelerated to maximum speed once they reached a channel. The statoliths accelerated sooner, and the channeling effect was less pronounced in roots treated with cytochalasin D. Parallel ultrastructural studies of high-pressure frozen-freeze-substituted columella cells suggest that the low-resistance statolith pathway in the cell periphery corresponds to the sharp interface between the endoplasmic reticulum (ER)-rich cortical and the ER-devoid central region of these cells. The central region is also shown to contain an actin-based cytoskeletal network in which the individual, straight, actin-like filaments are randomly distributed. To explain these findings as well as the results of physical simulation experiments, we have formulated a new, tensegrity-based model of gravity sensing in columella cells. This model envisages the cytoplasm as pervaded by an actin-based cytoskeletal network that is denser in the ER-devoid central region than in the ER-rich cell cortex and is linked to stretch receptors in the plasma membrane. Sedimenting statoliths are postulated to produce a directional signal by locally disrupting the network and thereby altering the balance of forces acting on the receptors in different plasma membrane regions. PMID:11161060

Amyloplast sedimentation dynamics in maize columella cells support a new model for the gravity-sensing apparatus of roots.

PubMed

Yoder, T L; Zheng, H Q; Todd, P; Staehelin, L A

2001-02-01

Quantitative analysis of statolith sedimentation behavior was accomplished using videomicroscopy of living columella cells of corn (Zea mays) roots, which displayed no systematic cytoplasmic streaming. Following 90 degrees rotation of the root, the statoliths moved downward along the distal wall and then spread out along the bottom with an average velocity of 1.7 microm min(-1). When statolith trajectories traversed the complete width or length of the cell, they initially moved horizontally toward channel-initiation sites and then moved vertically through the channels to the lower side of the reoriented cell where they again dispersed. These statoliths exhibited a significantly lower average velocity than those sedimenting on distal-to-side trajectories. In addition, although statoliths undergoing distal-to-side sedimentation began at their highest velocity and slowed monotonically as they approached the lower cell membrane, statoliths crossing the cell's central region remained slow initially and accelerated to maximum speed once they reached a channel. The statoliths accelerated sooner, and the channeling effect was less pronounced in roots treated with cytochalasin D. Parallel ultrastructural studies of high-pressure frozen-freeze-substituted columella cells suggest that the low-resistance statolith pathway in the cell periphery corresponds to the sharp interface between the endoplasmic reticulum (ER)-rich cortical and the ER-devoid central region of these cells. The central region is also shown to contain an actin-based cytoskeletal network in which the individual, straight, actin-like filaments are randomly distributed. To explain these findings as well as the results of physical simulation experiments, we have formulated a new, tensegrity-based model of gravity sensing in columella cells. This model envisages the cytoplasm as pervaded by an actin-based cytoskeletal network that is denser in the ER-devoid central region than in the ER-rich cell cortex and is linked to stretch receptors in the plasma membrane. Sedimenting statoliths are postulated to produce a directional signal by locally disrupting the network and thereby altering the balance of forces acting on the receptors in different plasma membrane regions.

Iron and ferritin accumulate in separate cellular locations in Phaseolus seeds

PubMed Central

2010-01-01

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 crop plants and they suggest that iron distribution criteria should be integrated into selection strategies for bean biofortification. PMID:20149228

From ROOTS to GRAVI-1: Twenty Five Years for Understanding How Plants Sense Gravity

NASA Astrophysics Data System (ADS)

Perbal, Gerald

2009-01-01

In the 1970s, when I started to work on gravitropism at the University Pierre and Marie Curie, Paris), it was well known that statocytes contain voluminous amyloplasts (statoliths) that sediment under the influence of gravity. The role of these organelles in gravisensing was strongly disputed. In 1974, I attended a session of a meeting on gravitropism in Würzburg, where I presented results that supported the involvement of statoliths in the perception of gravity. This meeting had a strong impact on my research, since at that time the Council of Europe was looking for people interested in performing experiments in Space. Our first experiment (ROOTS) was carried out in the Biorack Facility (ESA) in the frame of the Spacelab D1 mission (1985). We had a very efficient help from CNES which developed a very fine hardware to grow lentil seedlings and to chemically fix them at the end of the experiment. The results obtained were surprising since we observed that in microgravity the statoliths were located at one pole of the statocyte and not distributed at random as it was expected. The goal of the following experiment (Spacelab IML-1 mission, 1992) was to determine the threshold stimulation time at 1 × g (created by centrifugation). It was estimated at 25 s. In the frame of the SMM/03 and SMM/06 missions (1996, 1997), we proved that the statoliths are attached to actin filaments by motor proteins (myosin) that make these organelles move in one preferential direction in microgravity. The analysis of gravisensitivity with clinostats incited us to compare gravisensitivity of lentil roots grown in microgravity or on a 1 × g centrifuge (SMM05 mission, 1997). It was found that the latter were less sensitive than the former. We showed that this was due to the fact that the statoliths are not distributed in the same way in both cases (microgravity or 1 × g centrifuge). All these studies led us to propose a mode of gravity sensing by plants in which elements of the cytoskeleton and stretch activated ion channels are involved. The last experiment (GRAVI-1) which has been carried out (in ISS with the EMCS facility, 2007) dealt with the threshold acceleration that is perceived by roots.

A temporary immersion system improves in vitro regeneration of peach palm through secondary somatic embryogenesis

PubMed Central

Steinmacher, D. A.; Guerra, M. P.; Saare-Surminski, K.; Lieberei, R.

2011-01-01

Background and Aims Secondary somatic embryogenesis has been postulated to occur during induction of peach palm somatic embryogenesis. In the present study this morphogenetic pathway is described and a protocol for the establishment of cycling cultures using a temporary immersion system (TIS) is presented. Methods Zygotic embryos were used as explants, and induction of somatic embryogenesis and plantlet growth were compared in TIS and solid culture medium. Light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to describe in vitro morphogenesis and accompany morpho-histological alterations during culture. Key Results The development of secondary somatic embryos occurs early during the induction of primary somatic embryos. Secondary somatic embryos were observed to develop continually in culture, resulting in non-synchronized development of these somatic embryos. Using these somatic embryos as explants allowed development of cycling cultures. Somatic embryos had high embryogenic potential (65·8 ± 3·0 to 86·2 ± 5·0 %) over the period tested. The use of a TIS greatly improved the number of somatic embryos obtained, as well as subsequent plantlet growth. Histological analyses showed that starch accumulation precedes the development of somatic embryos, and that these cells presented high nucleus/cytoplasm ratios and high mitotic indices, as evidenced by DAPI staining. Morphological and SEM observations revealed clusters of somatic embryos on one part of the explants, while other parts grew further, resulting in callus tissue. A multicellular origin of the secondary somatic embryos is hypothesized. Cells in the vicinity of callus accumulated large amounts of phenolic substances in their vacuoles. TEM revealed that these cells are metabolically very active, with the presence of numerous mitochondria and Golgi apparatuses. Light microscopy and TEM of the embryogenic sector revealed cells with numerous amyloplasts, large nuclei and nucleoli, and numerous plasmodesmata. Plantlets were obtained and after 3 months in culture their growth was significantly better in TIS than on solid culture medium. However, during acclimatization the survival rate of TIS-grown plantlets was lower. Conclusions The present study confirms the occurrence of secondary somatic embryos in peach palm and describes a feasible protocol for regeneration of peach palm in vitro. Further optimizations include the use of explants obtained from adult palms and improvement of somatic embryo conversion rates. PMID:21355009

Actin Cytoskeleton-Based Plant Synapse as Gravitransducer in the Transition Zone of the Root Apex

NASA Astrophysics Data System (ADS)

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, Volkmann D, Barlow PW (2009b) The 'root-brain' hypothesis of Charles and Francis Darwin: Revival after more than 125 years. Plant Signal Behav 4: 1121-1127 Mancuso S, Barlow PW, Volkmann D, Balǔka F (2006). Actin turnover-mediated gravity response in s maize root apices: gravitropism of decapped roots implicates gravisensing outside of the root cap. Plant Signal Behav 1: 52-58

Identification and characterization of plastid-type proteins from sequence-attributed features using machine learning

PubMed Central

2013-01-01

Background Plastids are an important component of plant cells, being the site of manufacture and storage of chemical compounds used by the cell, and contain pigments such as those used in photosynthesis, starch synthesis/storage, cell color etc. They are essential organelles of the plant cell, also present in algae. Recent advances in genomic technology and sequencing efforts is generating a huge amount of DNA sequence data every day. The predicted proteome of these genomes needs annotation at a faster pace. In view of this, one such annotation need is to develop an automated system that can distinguish between plastid and non-plastid proteins accurately, and further classify plastid-types based on their functionality. We compared the amino acid compositions of plastid proteins with those of non-plastid ones and found significant differences, which were used as a basis to develop various feature-based prediction models using similarity-search and machine learning. Results In this study, we developed separate Support Vector Machine (SVM) trained classifiers for characterizing the plastids in two steps: first distinguishing the plastid vs. non-plastid proteins, and then classifying the identified plastids into their various types based on their function (chloroplast, chromoplast, etioplast, and amyloplast). Five diverse protein features: amino acid composition, dipeptide composition, the pseudo amino acid composition, Nterminal-Center-Cterminal composition and the protein physicochemical properties are used to develop SVM models. Overall, the dipeptide composition-based module shows the best performance with an accuracy of 86.80% and Matthews Correlation Coefficient (MCC) of 0.74 in phase-I and 78.60% with a MCC of 0.44 in phase-II. On independent test data, this model also performs better with an overall accuracy of 76.58% and 74.97% in phase-I and phase-II, respectively. The similarity-based PSI-BLAST module shows very low performance with about 50% prediction accuracy for distinguishing plastid vs. non-plastids and only 20% in classifying various plastid-types, indicating the need and importance of machine learning algorithms. Conclusion The current work is a first attempt to develop a methodology for classifying various plastid-type proteins. The prediction modules have also been made available as a web tool, PLpred available at http://bioinfo.okstate.edu/PLpred/ for real time identification/characterization. We believe this tool will be very useful in the functional annotation of various genomes. PMID:24266945

ARG1 and ARL2 contribute to gravity signal transduction in the statocytes of Arabidopsis thaliana roots and hypocotyls

NASA Astrophysics Data System (ADS)

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

Gravity is an important cue that plant organs use to guide their growth. Each organ is characterized by a defined gravity set point angle that dictates its optimal orientation within the gravity field. Specialized cells, named statocytes, enable this directional growth response by perceiving gravity via the sedimentation of, and/or tension/pressure exerted by, starch-filled plastids within their cytoplasm. Located in the columella region of the cap in roots and in the endodermis of hypocotyls and stems, these cells modulate the lateral transport of auxin across the corresponding organ in a gravistimulus-dependent manner. Upon plant reorientation within the gravity field, a gravity signal transduction pathway is activated within those cells, which in roots leads to a relocalization of the PIN3 auxin efflux carrier toward the lower membrane and an alkalinization of the cytoplasm. In turn, these events appear to promote a lateral transport of auxin toward the bottom side of the stimulated organ, which promotes a curvature. We previously uncovered ARG1 and ARL2 as essential contributors to these cellular processes. Mutations in these genes result in altered root and hypocotyl gravitropism. In roots, this abnormal growth behavior is associated with a lack of PIN3 relocalization within the statocytes and an absence of preferential downward auxin transport upon gravistimulation. These two genes encode paralogous J-domain proteins that are associated with the plasma membrane and other membranes of the vesicular trafficking pathway, and appear to modulate protein trafficking within the statocytes. An analysis of the root gravitropic phenotypes associated with different double mutant configurations affecting ARG1, ARL2 and PIN3 suggest that all three proteins function in a common gravity-signaling pathway. Surprisingly, when a mutation that affects starch biosynthesis (pgm) is introgressed into an arg1-2 mutant, the gravitropic defects are dramatically enhanced relative to 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.

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

NASA Astrophysics Data System (ADS)

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

2014-05-01

Thermo Electric Power Plants (TEPP) produce a high amount of ash, that contains heavy metals and radionuclides. Ash is usually stored in ash-pits, in mixture with water and contains U235, Th 234 and their decay products, that are released from the coal matrix, during combustion, as well as heavy metals. Warm weather dried the ash and it can be spread by the wind in surrounded area. This paper presents the results of an experiment with two Salix genotypes, cultivated on an old closed ash-pit, nearby the Rovinari TEPP, in the middle Jiu valley (Gorj District, Romania), in order to evaluate its tolerance to heavy metals and radionuclides. Ash analysis revealed the presence of natural radionuclides, beloging from ash and coal dust, as well as of Cs 137, of Chernobil provenance. Radionuclides content over the normal limits for Romania were registered for Th 234, Pb210, U235 and Ra226. The heavy metals level in ash was over the normal limits, but under the alerts limits. In order to establish the woody plants tolerance to heavy metals and radionuclides, it is important to study the seedlings behavior. In this respect Salix alba and Salix viminalis whips and cuttings culture have been establish on Rovinari ash-pit. The observations made on survival and growth rate pointed out the superiority of Salix viminalis behaviour. After a period of three years Salix viminalis registered a 96% survival rate, while in Salix alba annual decreases, reaching to 14%. These results are supported by the radionuclides content in leaves and by the electron microscopy studies. In Salix alba the leaves parenchimatic cells present a low sinthesis activity. The exogenous particles are accumulated in parenchima cells vacuola, the chloroplasts are usually agranal, with few starch grains and mitocondria presents slightly dillated crista. The ultrastructural features of the mature leaf cells, evidenced the natural adaptation of Salix viminalins for development in an environment with a big amount of 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.

Water mediated alterations in gravity signal transform phytofilertation capability in hydroponic plants

NASA Astrophysics Data System (ADS)

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

2012-07-01

An exorbitant sum of different synthetic molecules of chemicals including dyes and pigments are discharged into the environment, mainly via industrial effluents every year worldwide. The physical-chemical treatments for remediation viz adsorption, precipitation, ion exchange or filtration have proved to be disadvantageous because of high cost, low efficiency and inapplicability to a wide variety of dyes, or the formation of by-products and thereby creating waste disposal problems. Similarly the limited ability of micro-organisms to degrade xenobiotic especially sulphonoaromatic compounds, limits the efficiency and, therefore, the use of conventional wastewater treatment plants. In this context, the development of alternative biological treatments to eliminate these pollutants from industrial effluents is an important requirement. Plant metabolism, is extremely diverse and can be exploited to treat recalcitrant pollutants, not degradable by bacteria or fungi and can act as an important global sink for environmental pollutants. The presence of putative metabolites, in leaves of hydrophytes has been observed, indicating the transformation of several xenobiotics. A diverse range of the enzymes involved in the early stages of the detoxification process are closely associated with the redox biochemistry of the cell. The activities of enzymes such as glutathione transferases, peroxidases and cytochrome P450 monooxygenases and its multigenic family have implications with respect to the maintenance of redox homeostasis. Besides activating xenobiotics, cytochromes P450 is involved vitally in cell signaling for counteracting buoyant balance. Signal transduction cascades, including the role of cytochrome P450 monooxygenases in responding to gravitational cues, appear to be affected by buoyancy as well. Gravitropism is the orientation of growth in response to gravity and involves the perception of the gravitational force in the columella cells of the root cap where the primary 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.

A weak combined magnetic field changes root gravitropism

NASA Astrophysics Data System (ADS)

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

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. According to the starch-statolith hypothesis, amyloplasts in the specialized graviperceptive cells - statocytes sediment in the direction of a gravitational vector in the distal part of a cell. The polar arrangement of organelles is maintained by means of the cytoskeleton. On the Kholodny-Went's, theory the root bending is provided by the polar movement of auxin from a root cap to the elongation zone. It is also known that gravistimulation initiates a rapid Ca2+ redistribution in a root apex. Calcium ions modify an activity of many cytoskeletal proteins and clustering of calcium channels may be directed by actin microfilaments. Although the available data show the Ca2+ and cytoskeleton participation in graviperception and signal transduction, the clear evidence with regard to the participation of cytoskeletal elements and calcium ions in these processes is therefore substantial but still circumstantial and requires new experimental data. Roots are characterized with positive gravitropism, i. e. they grow in the direction of a gravitational vector. It was first shown by us that roots change the direction of a gravitropic reaction under gravistimulation in the weak combined magnetic field with a frequency of 32 Hz. 2-3-day old cress seedlings were gravistimulated in moist chambers, which are placed in μ-metal shields. Inside μ -metal shields, combined magnetic fields have been created. Experiments were performed in darkness at temperature 20±10C. Measurements of the magnitude of magnetic fields were carried out with a flux-gate magnetometer. Cress roots reveal negative gravitropism, i. e. they grow in the opposite direction to a gravitational vector, during 2 h of gravistimulation and then roots begin to grow more or less parallel to the Earth's surface, i.e. they reveal plagiotropism. Since such combined magnetic field is adjusted to the cyclotron frequency of Ca2+ ions, these observations demonstrate the participation of calcium ions in root gravitropism. Cyclotron frequency of Ca2+ ions is the formal frequency of ion rotation in the static magnetic field. Simultaneous applying the altering magnetic field with the same frequency can provoke auto-oscillation in the system and consequently change the rate and/or the direction of Ca2+ ion flow in a root under gravistimulation. The data of light, electron, and confocal laser microscopy and kinetics of a gravitropic reaction, which have been obtained on such the new original model, are discussed in the light of current concepts of root gravitropism.

Reminiscences, collaborations and reflections.

PubMed

Akazawa, T

1994-02-01

This is a personal account by a semi old-timer who completed his official term as a professor of plant biochemistry at Nagoya University in Japan in 1992. My university student life began soon after the World War II (1948). I shared the hardships of many in my age group, in that life was difficult during my college years. I was fortunate to have the opportunity of studying in the USA on a Fulbright scholarship first at Purdue University (1955-1956), and then at the University of California, Berkeley (1956-1957). My graduate study and postdoctoral training in the new world were vitally refreshing and stimulating, which gave me the impetus for becoming a natural scientist associated with academic institutions. Consciously and subconsciously I was impressed by the friendly and liberal atmosphere surrounding young students as well as senior scholars in the United States. But more importantly, I was inspired by the critical and competitive minds prevailing among these people.The appointment as a biochemist at the International Rice Research Institute (IRRI) in the Philippines (1962-1964) was the real start of my professional career. The work was continued upon my return to Nagoya to become a staff member of the Research Institute for Biochemical Regulation (1964-1992). Throughout the years, my major research interest has covered photosynthesis as a whole, involving photosynthetic CO2-fixation (RuBisCO), carbohydrate metabolism, e.g. starch biosynthesis and breakdown (α-amylase), and metabolic regulation, which are interrelated in the basic metabolism of plant cells.I shall briefly describe in this article highlights from my studies and discoveries made and I shall also discuss their possible significance in plant metabolism, with the hope that it does not contradict my sense of humility: They are (a) discovery of ADPG in plants and its role in starch biosynthesis; (b) structure-function relationship of RuBisCO proteins, in particular on heterologous recombination of their subunits of plant-type enzyme molecules derived from the prokaryotic photosynthetic bacteria; (c) molecular evolution of RuBisCO genes; (d) mode of actions (formation, intracellular transport and secretion) of rice seed α-amylase and its structural characteristics (distinctive glycosylation), and (e) DNA methylation and regulatory mechanism of photosynthesis gene expression in plastids (amyloplasts). In each step of my research, I shared joy, excitement, disappointment, and agony with my colleagues, an experience that may be common to all researchers. Although it is now becoming well recognized among the scientific community in Japan, I want to point out that interaction of multinational scientific minds in the laboratory produces a vital and creative atmosphere for performance of successful research. I experienced and realized this important fact in my earlier days in the USA and the Philippines. Inasmuch as I believe that this is the most crucial element for any research laboratory to possess, I fondly remember the friendships gained with numerous overseas visitors and collaborators who have contributed immensely to our work.

``Rhizogenesis in vitro'' - as a model to study microgravity biological effects

NASA Astrophysics Data System (ADS)

Bulavin, Iliya

Functioning organisms is based on the physiological and biochemical processes in different tissues and cells. Numerous spaceflight biological experiments have shown the essential changes in cell behavior of multicellular and unicellular organisms in comparison with that on Earth. In our investigations, we used the model “Rhizogenesis in vitro” to study cell differentiation in the root cap and growth zones under clinorotation. Advantage of this model is the possibility to study the influence of clinorotation at the beginning of root initiation de novo and next morphogenetic processes unlike experiments in vivo with embryonal seedling roots formed in seeds. Arabidopsis thaliana plants of wild type and scr mutant (3999 by NASC database) were used. For rhizogenesis induction, rosette leaves with petioles were cut and transferred in Petri dishes on MS medium contained 1/10 of MS mineral salt, without vitamins and hormones. One half of Petri dishes were placed vertically (control), the other - on a slow horizontal clinostat (2 rpm). Anatomical investigation of A. thaliana wild type and scr mutant roots formed de novo showed that formation of root cap and growth zones (meristem, distal elongation zone (DEZ), central elongation zone (CEZ) and mature zone) under clinorotation was similar to that in control. A root cap consists of columella and peripheral cells. In the columella there are meristematic cells, statocytes (graviperceptive cells), and secretory cells. Epidermis, parenchyma, endodermis and central cylinder are distinguished in wild type roots. Unlike a wild type, a cortex of scr mutant was represented by one cell layer which had the parenchyma and endodermis characteristics. A root cap length and width were similar in control and under clinorotation. A cell number in the meristem and DEZ and a length of these growth zones did not differ in control and the experimental conditions. The ultrasructure of cap meristematic cells was typical for cells of this 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.