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Genotypic and developmental evidence for the role of plasmodesmatal regulation in cotton fiber elongation mediated by callose turnover.

Cotton fibers are single-celled hairs that elongate to several centimeters long from the seed coat epidermis of the tetraploid species (Gossypium hirsutum and Gossypium barbadense). Thus, cotton fiber is a unique system to study the mechanisms of rapid cell expansion. Previous work has shown a transient closure of plasmodesmata during fiber elongation (Y.-L. Ruan, D.J. Llewellyn, R.T. Furbank [2001] Plant Cell 13: 47-60). To examine the importance of this closure in fiber elongation, we compared the duration of the plasmodesmata closure among different cotton genotypes differing in fiber length. Confocal imaging of the membrane-impermeant fluorescent molecule carboxyfluorescein revealed a genotypic difference in the duration of the plasmodesmata closure that positively correlates with fiber length among three tetraploid genotypes and two diploid progenitors. In all cases, the closure occurred at the rapid phase of elongation. Aniline blue staining and immunolocalization studies showed that callose deposition and degradation at the fiber base correlates with the timing of plasmodesmata closure and reopening, respectively. Northern analyses showed that the expression of a fiber-specific beta-1,3-glucanase gene, GhGluc1, was undetectable when callose was deposited at the fiber base but became evident at the time of callose degradation. Genotypically, the level of GhGluc1 expression was high in the short fiber genotype and weak in the intermediate and long fiber genotypes. The data provide genotypic and developmental evidence that (1) plasmodesmata closure appears to play an important role in elongating cotton fibers, (2) callose deposition and degradation may be involved in the plasmodesmata closure and reopening, respectively, and (3) the expression of GhGluc1 could play a role in this process by degrading callose, thus opening the plasmodesmata.

Cell Enlargement↗

Cellulose and Callose Biosynthesis in Higher Plants (I. Solubilization and Separation of (1->3)- and (1->4)-[beta]-Glucan Synthase Activities from Mung Bean).

(1->3)- and (1->4)-[beta]-glucan synthase activities from higher plants have been physically separated by gel electrophoresis in nondenaturing conditions. The two glucan synthases show different mobilities in native polyacrylamide gels. Further separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a different polypeptide composition in these synthases. Three polypeptides (64, 54, and 32 kD) seem to be common to both synthase activities, whereas two polypeptides (78 and 38 kD) are associated only with callose synthase activity. Twelve polypeptides (170, 136, 108, 96, 83, 72, 66, 60, 52, 48, 42, and 34 kD) appear to be specifically associated with cellulose synthase activity. The successful separation of (1->3)- and (1->-4)-[beta]-glucan synthase activities was based on the manipulation of digitonin concentrations used in the solubilization of membrane proteins. At low dipitomin concentrations (0.05 and 0.1%), the ratio of the cellulose to callose synthase activity was higher. At higher digitonin (0.5-1%) concentrations, the ratio of the callose to cellulose synthase activity was higher. Rosette-like particles with attached product were observed in samples taken from the top of the stacking gel, where only cellulose was synthesized. Smaller (nonrosette) particles were found in the running gel, where only callose was synthesized. These findings suggest that a higher level of subunit organization is required for in vitro cellulose synthesis in comparison with callose assembly.

Journal Article↗

Callose deposition is responsible for apoplastic semipermeability of the endosperm envelope of muskmelon seeds

Semipermeable cell walls or apoplastic "membranes" have been hypothesized to be present in various plant tissues. Although often associated with suberized or lignified walls, the wall component that confers osmotic semipermeability is not known. In muskmelon (Cucumis melo L.) seeds, a thin, membranous endosperm completely encloses the embryo, creating a semipermeable apoplastic envelope. When dead muskmelon seeds are allowed to imbibe, solutes leaking from the embryo are retained within the envelope, resulting in osmotic water uptake and swelling called osmotic distention (OD). The endosperm envelope of muskmelon seeds stained with aniline blue, which is specific for callose (beta-1,3-glucan). Outside of the aniline-blue-stained layer was a Sudan III- and IV-staining (lipid-containing) layer. In young developing seeds 25 d after anthesis (DAA) that did not exhibit OD, the lipid layer was already present but callose had not been deposited. At 35 DAA, callose was detected as distinct vesicles or globules in the endosperm envelope. A thick callose layer was evident at 40 DAA, coinciding with development of the capacity for OD. Removal of the outer lipid layer by brief chloroform treatment resulted in more rapid water uptake by both viable and nonviable (boiled) seeds, but did not affect semipermeability of the endosperm envelope. The aniline-blue-staining layer was digested by beta-1,3-glucanase, and these envelopes lost OD. Thus, apoplastic semipermeability of the muskmelon endosperm envelope is dependent on the deposition of a thick callose-containing layer outside of the endosperm cell walls.

Journal Article↗

Callose Deposition and Photoassimilate Export in Phaseolus vulgaris Exposed to Excess Cobalt, Nickel, and Zinc.

Callose accumulated on sieve plates of phloem of white bean seedlings exposed to excess Co, Ni, or Zn. The callose deposits ranged in thickness and were most pronounced in midribs of unifoliate leaves and their subtending petioles. Lesser callose deposits were found in stems. Although translocation of (14)C was reduced drastically in seedlings exposed to excess metal, no correlation was found between translocated (14)C and the amount of callose in the petioles. It is concluded that the inhibition of phloem translocation in seedlings exposed to excess metal is due to effects other than callose deposition.

Journal Article↗

Direct Photolabeling with [P]UDP-Glucose for Identification of a Subunit of Cotton Fiber Callose Synthase.

We have identified a 52 kilodalton polypeptide as being a likely candidate for the catalytic subunit of the UDP-glucose: (1-->3)-beta-glucan (callose) synthase of developing fibers of Gossypium hirsutum (cotton). Such a polypeptide migrates coincident with callose synthase during glycerol gradient centrifugation in the presence of EDTA, and can be directly photolabeled with the radioactive substrate, alpha-[(32)P]UDP-glucose. Interaction with the labeled probe requires Ca(2+), a specific activator of callose synthase which is known to lower the K(m) of higher plant callose synthases for the substrate UDP-glucose. Using this probe and several other related ones, several other proteins which interact with UDP-glucose were also identified, but none satisfied all of the above criteria for being components of the callose synthase.

Journal Article↗

Premature dissolution of the microsporocyte callose wall causes male sterility in transgenic tobacco.

Male sterility in a petunia cytoplasmic male sterile line has been attributed to the early appearance of active callase, a beta-1,3-glucanase, in the anther locule. This leads to premature dissolution of the callose walls surrounding the microsporogenous cells. We have mimicked this aspect of the petunia line in transgenic tobacco by engineering the secretion of a modified pathogenesis-related vacuolar beta-1,3-glucanase from the tapetum prior to the appearance of callase activity in the locule. Plants expressing the modified glucanase from tapetum-specific promoters exhibited reduced male fertility, ranging from complete to partial male sterility. Callose appearance and distribution are normal in the male sterile transgenic plants up to prophase I, whereupon callose is prematurely degraded. Meiosis and cell division occur normally. The resultant microspores have an abnormally thin cell wall that lacks sculpturing. The tapetum shows hypertrophy. Male sterility is probably caused by bursting of the aberrant microspores at a time corresponding to microspore release. These results demonstrate that premature callose degradation is sufficient to cause male sterility and suggest that callose is essential for the formation of a normal microspore cell wall.

Base Sequence↗

Conflict of intentions due to callosal disconnection.

OBJECTIVES: Three patients with callosal syndrome manifested a peculiar symptom in that they were unable to perform intended whole body actions because another intention emerged in competition with the original one. Attempts were made to clarify the symptomatology of this manifestation and its possible mechanism is discussed. METHODS: The three patients are described and previous reports on patients with callosal damage were reviewed. Four additional patients with similar symptoms were found and the clinical features common to all seven patients were examined. RESULTS: This symptom could not be attributed to unilateral movement disorders such as unilateral apraxia, intermanual conflict, or compulsive manipulation of tools. The manifestations included marked hesitation in initiating actions, interruption of actions, repetitive actions, and performance of unintended actions with difficulty in correcting them. All patients, except one, had a lesion in the posterior half of the body of the corpus callosum, and there was no significant involvement of the cerebral cortex. The symptom became manifest later than 4 weeks after callosal damage. It occurred during spontaneous actions, but not during well automated actions nor when following instructions. CONCLUSION: This symptom, tentatively named "conflict of intentions", can be regarded as a fragment of diagonistic dyspraxia originally described by Akelaitis, although it can occur independently of intermanual conflict. Normally, the right and left cerebral hemispheres may be complementarily modifying automated whole body actions in order to adapt behaviour to changes of the environment as well as to the intention. Partial callosal disconnection without significant cortical involvement would exaggerate the disparity between the role of each hemisphere through the reorganisation of neural systems after callosal damage. Such double, often contrary, behavioural tendencies may sometimes simultaneously enter the patient's awareness.

Adult↗

Synaptic enhancement induced through callosal pathways in cat association cortex.

The corpus callosum plays a major role in synchronizing neocortical activities in the two hemispheres. We investigated the changes in callosally elicited excitatory postsynaptic potentials (EPSPs) of neurons from cortical association areas 5 and 7 of cats under barbiturate or ketamine-xylazine anesthesia. Single pulses to callosal pathway evoked control EPSPs; pulse-trains were subsequently applied at different frequencies to homotopic sites in the contralateral cortex, as conditioning stimulation; thereafter, the single pulses were applied again to test changes in synaptic responsiveness by comparing the amplitudes of control and conditioned EPSPs. In 41 of 42 neurons recorded under barbiturate anesthesia, all frequencies of conditioning callosal stimuli induced short-term (5-30 min) enhancement of test EPSPs elicited by single stimuli. Neurons tested with successive conditioning pulse-trains at different frequencies displayed stronger enhancement with high-frequency (40-100 Hz) than with low-frequency (10-20 Hz) rhythmic pulse-trains; >100 Hz, the potentiation saturated. In a neuronal sample, microdialysis of an N-methyl-D-aspartate (NMDA) receptor blocker in barbiturate-treated cats suppressed this potentiation, and potentiation of callosally evoked EPSPs was not detected in neurons recorded under ketamine-xylazine anesthesia, thus indicating that EPSPs' potentiation implicates, at least partially, NMDA receptors. These data suggest that callosal activities occurring within low-frequency and fast-frequency oscillations play a role in cortical synaptic plasticity.

2-Amino-5-phosphonovalerate↗

The PTI1-like kinase ZmPti1a from maize (Zea mays L.) co-localizes with callose at the plasma membrane of pollen and facilitates a competitive advantage to the male gametophyte.

BACKGROUND: The tomato kinase Pto confers resistance to bacterial speck disease caused by Pseudomonas syringae pv. tomato in a gene for gene manner. Upon recognition of specific avirulence factors the Pto kinase activates multiple signal transduction pathways culminating in induction of pathogen defense. The soluble cytoplasmic serine/threonine kinase Pti1 is one target of Pto phosphorylation and is involved in the hypersensitive response (HR) reaction. However, a clear role of Pti1 in plant pathogen resistance is uncertain. So far, no Pti1 homologues from monocotyledonous species have been studied. RESULTS: Here we report the identification and molecular analysis of four Pti1-like kinases from maize (ZmPti1a, -b, -c, -d). These kinase genes showed tissue-specific expression and their corresponding proteins were targeted to different cellular compartments. Sequence similarity, expression pattern and cellular localization of ZmPti1b suggested that this gene is a putative orthologue of Pti1 from tomato. In contrast, ZmPti1a was specifically expressed in pollen and sequestered to the plasma membrane, evidently owing to N-terminal modification by myristoylation and/or S-acylation. The ZmPti1a:GFP fusion protein was not evenly distributed at the pollen plasma membrane but accumulated as an annulus-like structure which co-localized with callose (1,3-beta-glucan) deposition. In addition, co-localization of ZmPti1a and callose was observed during stages of pollen mitosis I and pollen tube germination. Maize plants in which ZmPti1a expression was silenced by RNA interference (RNAi) produced pollen with decreased competitive ability. Hence, our data provide evidence that ZmPti1a plays an important part in a signalling pathway that accelerates pollen performance and male fitness. CONCLUSION: ZmPti1a from maize is involved in pollen-specific processes during the progamic phase of reproduction, probably in crucial signalling processes associated with regions of callose deposition. Pollen-sporophyte interactions and pathogen induced HR show certain similarities. For example, HR has been shown to be associated with cell wall reinforcement through callose deposition. Hence, it is hypothesized that Pti1 kinases from maize act as general components in evolutionary conserved signalling processes associated with callose, however during different developmental programs and in different tissue types.

Cell Membrane↗

Dissociation of letter and picture naming resulting from callosal disconnection.

BACKGROUND: Detailed mapping of the corpus callosum for functional fractionation in humans remains incomplete. OBJECTIVE: To examine separable interhemispheric transfer of visual information by callosal fibers, especially in the splenium. METHODS: We examined callosal disconnection signs in a 14-year-old boy with a lesion confined to the posterior part of the splenium and reviewed reported cases with callosal lesions. RESULTS AND CONCLUSION: The patient presented with left hemialexia as the only manifestation of callosal disconnection syndrome. The only difficulty demonstrated was in reading aloud or copying letters, which were presented tachistoscopically to the left visual field, with his right hand. He could copy letters presented to his left visual field with his left hand, however. Therefore, left hemialexia was not due to hemiamblyopia or hemineglect. There was no anomia for pictures and colors in the left visual field. MRI revealed that the lesion was limited to the ventroposterior end of the splenium. Review of 40 reported patients with callosal lesions suggests that the anterior to middle part of the splenium is involved in transferring picture information from the language-nondominant hemisphere to the language-dominant hemisphere and that the ventroposterior part is involved in transferring letter information.

Adolescent↗

Callose biosynthesis as a Ca2+-regulated process and possible relations to the induction of other metabolic changes.

Chitosan and poly-L-lysine induce electrolyte leakage in suspension-cultured soy bean cells due to their polycationic nature. Similar effects are caused by Polymyxin B at certain concentrations. After any of these treatments electrolyte leakage is followed by a rapid onset of callose synthesis, as studied quantitatively by its fluorescence with Aniline Blue. In addition to membrane perturbations resulting in membrane leakage callose synthesis requires the presence of external Ca2+ at microM concentrations. It is suggested that concomitant with electrolyte leakage a Ca2+ influx occurs, which leads to a local increase in Ca2+ at the cytoplasmic side of the membrane, resulting in activation of the (1----3)-beta-glucan synthase. The activity of this enzyme in microsomal preparations depends directly and reversibly on Ca2+, with half-saturation at about 5 microM. Inhibitor studies suggest that activation of the (1----3)-beta-glucan synthase by Ca2+ is not mediated by calmodulin but possibly by phospholipids. The enzyme can also be activated in vitro by limited proteolysis. However, this type of activation does not appear to be involved in chitosan-elicited callose formation. It appears of interest in regard to the function of callose in cell and tissue repair mechanisms that its formation is inhibited by unsaturated fatty acids and lysophosphatidylcholine, substances that presumably can result during membrane damage. Callose synthesis may be regarded as an indicator of chitosan-induced increase in cytoplasmic Ca2+ concentration. It is speculated that this may also be part of the signal chain for initiation of de novo synthesis of enzymes reported to regulate phytoalexin production.

Calcium↗

Post-pollination callose development in ovules of Rhododendron and Ledum (Ericaceae): zygote special wall.

In Rhododendron spp. and Ledum groenlandicum a callose wall is laid down around the zygote in the first 2 days after fertilization. The periodic acid/Schiff-positive, aniline blue-fluorescence-positive callosic wall is initiated adjacent to the degenerating synergid, extends to cover the entire zygote surface, and remains visible during the initiation of embryogeny as the zygote elongates before the first proembryonal division. Unfertilized ovules show eventual callose deposition in the ovule wall cells during senescence in undeveloped abscising pistils, but show no development of callose within the embryo sac. Possible roles of a zygote special callose wall are discussed.

Cell Wall↗

Fluorochromes for detection of callose in meiocytes of olive (Olea europaea L.).

The callosic wall which covers microsporocyte mother cells during meiotic division has been studied using different fluorochromes as alternatives to the widely used aniline blue. We have confirmed that both acridine orange and 4', 6' diamidino-2-phenylindole (DAPI) produce a fluorescent response to callose which is comparable in specificity and intensity to that of aniline blue; therefore, they can be used to study callose wall formation. Staining properties of these fluorochromes, as well of those of curcumin and sirofluor, reported earlier as fluorescent stains for callose, are discussed. We also discuss the efficacy of the combined use of sirofluor and DAPI to study particular aspects of the deposition of callose.

Coloring Agents↗

Corpus callosal atrophy following closed head injury: detection with magnetic resonance imaging.

To investigate evidence for diffuse white matter injury and hemispheric disconnection sequelae after severe closed head injury (CHI), this study evaluates the degree of posttraumatic atrophy of the corpus callosum. Corpus callosal atrophy was quantitatively determined using a digitizer to measure sagittal magnetic resonance images of 32 patients with moderate-to-severe CHI and those of 31 control subjects of similar age. In the CHI patients, measurements were significantly reduced for the areas of the anterior four-fifths, the posterior one-fifth, and the total corpus callosum. Moreover, the minimum width of the callosal body was reduced in the CHI patients as compared to that of control individuals. Indices of corpus callosal atrophy were significantly correlated with the chronicity of injury and the degree of lateral ventricular enlargement. There was no difference in callosal measurements between men and women. Magnetic resonance imaging provides an in vivo determination of corpus callosal atrophy which may reflect the severity of diffuse axonal injury and predict the type and severity of hemispheric disconnection effects.

Adolescent↗

[Callosal dementia: behavioral disorders related to central and extrapontine myelinolysis].

We report the behavioral symptoms presented by a 57-year-old man as the first sign of a Marchiafava-Bignami syndrome and by a 44-year-old woman with centro and extrapontine myelinolysis. These observations define a clinical entity, that we named callosal dementia characterized by: 5) fronto-limbic signs with coarse interjections, repetitive and antisocial behavior, alternation of lack of incitation and agitation; 6) elements of a Balint syndrome (suggestive of a posterior callosal involvement), with a pseudo-hallucinated look and a gaze apraxia; 7) signs of callosal dysconnection and; signs of adjacent white matter involvement, with paucity of vocal and facial expression modulation. Early recognition of these features of callosal dementia may be very helpful for diagnosis of suspected myelinolysis, leading to a more careful research of clinical signs of callosal dysconnection and prompting neuroimaging with MRI. A rapid confirmation of the diagnosis may prevent progression to centro or extrapontine myelinolysis, that may sometimes still be lethal, by adequate supportive measures (slow correction of electrolytes imbalance, correction of deficiencies, total alcohol withdrawal).

Adult↗

Functional specificity of callosal connections in tree shrew striate cortex.

Although callosal connections have been shown to link extensive regions of primary visual cortex, the distribution of these connections with respect to the map of visual space and the map of orientation preference remains unclear. Here we combine optical imaging of intrinsic signals with injection of fluorescent microspheres to assess the functional specificity of callosal connections in the tree shrew. By imaging both hemispheres simultaneously while presenting a series of spatially restricted stimuli, we find that a substantial region of visual space is represented bilaterally. Each hemisphere includes a representation of the ipsilateral visual field that is highly compressed relative to that of the contralateral visual field and is most extensive in the lower visual field, where approximately 30(o) of central visual space are represented bilaterally. Callosal connections extend throughout the region of bilateral representation but terminate in a spatially restricted manner that links visuotopically corresponding sites in the two hemispheres. In contrast, callosal connections appear to terminate without regard for the map of orientation preference, showing little sign of the orientation-specific modular and axial specificity that is characteristic of long-range horizontal connections. By coordinating the activity in the two hemispheres in a way that preserves nearest neighbor relationships, callosal connections may best be viewed as elements of local circuits that operate within a single bilateral representation of visual space.

Animals↗

The association between stress, hemispheric specialization, and callosal interactions.

This study examines the effects of stress on hemispheric specialization and callosal transfer. Previous research suggests that the two cerebral hemispheres are asymmetric in their regulation of and response to stress. Further, studies have suggested that the activity of the corpus callosum may also be affected by stress. A group of 28 participants completed the Spielberger State-Trait Anxiety Inventory and performed a bilateral Stroop task. The bilateral Stroop task contains separate conditions which measure both hemispheric specialization and callosal transfer. A significant correlation was found between state anxiety and our measure of callosal transfer. Specifically, those individuals with greater state anxiety experience more efficient callosal transfer than did those with lower state anxiety. We conclude that state anxiety enhances callosal transfer without affecting hemispheric specialization.

Adolescent↗

Apparent atypical callosal dysgenesis: analysis of MR findings in six cases and their relationship to holoprosencephaly.

The MR scans of six pediatric patients with apparent atypical callosal dysgenesis (presence of the dorsal corpus callosum in the absence of a rostral corpus callosum) were critically analyzed and correlated with developmental information in order to assess the anatomic, embryologic, and developmental implications of this unusual anomaly. Four patients had semilobar holoprosencephaly; the dorsal interhemispheric commissure in these four infants resembled a true callosal splenium. All patients in this group had severe developmental delay. The other two patients had complete callosal agenesis with an enlarged hippocampal commissure mimicking a callosal splenium; both were developmentally and neurologically normal. The embryologic implications of the presence of these atypical interhemispheric connections are discussed. Differentiation between semilobar holoprosencephaly and agenesis of the corpus callosum with enlarged hippocampal commissure--two types of apparent atypical callosal dysgenesis--can be made by obtaining coronal, short TR/TE MR images through the frontal lobes. Such differentiation has critical prognostic implications.

Abnormalities, Multiple↗