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PubMed2006

Altered tapetal PCD and pollen wall development in the Arabidopsis ms1 mutant.

The Arabidopsis male sterility1 mutation results in mature anthers that are devoid of pollen. Meiosis and early development progress normally; however, after microspore release, the microspore cytoplasm and tapetum become abnormally granular and vacuolated, and degeneration occurs. Pollen wall development is seriously affected; primexine formation within the callose wall appears to occur normally, however, once the callose is degraded, abnormal deposits of electrodense material are detected which result in irregular spike-shaped structures, rather than the characteristic rod-like shape of the wild-type bacula. The internal intine wall is also reduced compared with wild type. TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling) staining and ultrastructural analysis have indicated that programmed cell death (PCD) occurs in the wild-type tapetum after microspore mitosis I. However, no signs of PCD are seen in the ms1 tapetum, where large autophagic vacuoles and mitochondrial swelling suggest that necrotic-based breakdown of the tapetum is occurring in the ms1 mutant rather than the normal, regulated PCD process. After the formation of the large, autophagic vacuole in the tapetum, TUNEL staining is detected in the mutant microspores, indicating that they may go through a PCD-based breakdown as a secondary consequence of the observed tapetal aberrations. Based on these observations, two possible roles for MS1 can be hypothesized; MS1 may function by modifying the transcription of tapetal-specific genes implicated in pollen wall development, which then regulate pollen wall material secretion and in turn wall development and tapetal PCD. Alternatively, the MS1 gene may control tapetal development by directly regulating tapetal PCD and breakdown.

Apoptosis↗
PubMed2006

Aluminium rhizotoxicity in maize grown in solutions with Al3+ or Al(OH)-4 as predominant solution Al species.

The rhizotoxicity of aluminium at low-pH with Al(3+) and at high pH with Al(OH)-(4) as the main Al species was studied. Aluminium reduced root growth to similar levels at pH 8.0 and pH 4.3, although the mononuclear Al concentration at pH 8.0 was three times lower than at pH 4.3. Al contents of root apices were much higher at pH 8 than at pH 4.3. Callose was induced only marginally at pH 8 and the formation was confined to the epidermis, whereas it proceeded through the cortex with time at pH 4.3. Well-documented genotypical differences in callose formation and Al accumulation could not be found at pH 8. The largest fraction of the root-tip Al was recovered in the cell-wall fraction independent of the solution pH. A sequential extraction of isolated cell walls suggests that most of the cell-wall Al was precipitated Al(OH)(3) at pH 8.0. This can be explained by a drastic pH reduction in the root apoplastic sap to 6.2, whereas at bulk solution pH 4.3 it rose to 5.6. Al precipitation was also confirmed by the microscopic localization of Al. At pH 8, Al could mostly be found in the epidermis, but in the apoplast of the outer cortex at pH 4.3. It is proposed here that at pH 4.3, Al(3+) inhibits root growth through binding to sensitive binding sites in the apoplast of the epidermis and the outer cortex. At pH 8, Al(OH)(3) precipitation in the epidermis causes a mechanical barrier thus impairing the root-growth control of the epidermis.

Aluminum↗
PubMed2004

Temporal and spatial distribution of pectin epitopes in differentiating anthers and microspores of fertile and sterile sugar beet.

We studied the possible involvement of several pectin epitopes in anther differentiation and microsporogenesis in fertile and cytoplasmically male sterile sugar beets. The spatial and temporal distribution of five structural motifs were traced with a panel of monoclonal antibodies in six stages: premeiosis, meiotic prophase, young and mature tetrads, young and expanding microspores. The composition of the walls of sporogenous cells and meiocytes differed than that in the tapetum, as evidenced by the presence of alpha-Fuc(1-->2)-beta-Gal and alpha-(1-->5)-L-Ara epitopes binding CCRC-M1 and LM6 antibodies. At meiotic prophase, the meiocyte walls were additionally marked by the appearance of poorly methyl-esterified domains of homogalacturonan and of (1-->4)-beta-Gal residues, detected by JIM5 and LM5. Some constituents of the meiocyte wall which reacted with JIM5 and JIM7 persisted on the surface of the special callose sheath during tetrad development. In newly formed primexine and exine layers of tetrads and microspores, epitopes that were bound by JIM5, JIM7 and LM5 were abundant. No differences in the deposition or relative abundance of pectins were found between fertile and sterile anthers until microspore release from the callose. Later, at the time of abortion, sterile microspores had much larger amounts of epitopes detected by JIM5 than their fertile counterparts.

Antibodies, Monoclonal↗
PubMed2006

Wall ingrowths in epidermal transfer cells of Vicia faba cotyledons are modified primary walls marked by localized accumulations of arabinogalactan proteins.

Despite the importance of transfer cells in enhancing nutrient transport in plants, little is known about how deposition of the complex morphology of their wall ingrowths is regulated. We probed thin sections of mature cotyledon epidermal transfer cells of Vicia faba with affinity probes and antibodies specific to polysaccharides and glycoproteins, to determine the distribution of these components in their walls. Walls of these transfer cells consist of the pre-existing primary wall, a uniformly deposited wall layer and wall ingrowths which are comprised of two regions; an electron-opaque inner region and an electron-translucent outer region. The primary wall reacted strongly with antibodies against esterified pectin, xyloglucan, the side chains of rhamnogalaturonan-1 and a cellulase-gold affinity probe. The electron-opaque inner region of wall ingrowths displayed a similar labeling pattern to that of the primary wall, showing strong cross-reactivity with all antibodies tested, except those reacting against highly de-esterified pectins. The electron-opaque outer layer of developmentally more mature wall ingrowths reacted strongly with anti-callose monoclonal and polyclonal antibodies, but showed no reaction for pectin or xyloglucan antibodies or the cellulase-gold affinity probe. The plasma membrane-wall interface was labeled strongly with anti-arabinogalactan protein (AGP) antibodies, with some AGP-reactive antibodies also labeling the electron-translucent zone. Nascent wall ingrowths were labeled specifically with AGPs but not anti-callose. A reduction in wall ingrowth density was observed when developing transfer cells were exposed to beta-d-glucosyl Yariv reagent compared with controls. Our results indicate that wall ingrowths of transfer cells are primary wall-like in composition and probably require AGPs for localized deposition.

Biological Transport↗
PubMed2003

Boron influences pollen germination and pollen tube growth in Picea meyeri.

To study the role of boron in pollen germination and pollen tube growth of Picea meyeri Rehd. et Wils., pollen grains were cultured in standard medium or boron-deficient medium. Effects of boron on the localization of pectins and callose in the walls of pollen tubes were observed by laser scanning confocal microscopy after staining with aniline blue or immunolabeling with antibodies JIM5 and JIM7. Changes in the structures of pectins and phenolics were investigated by fourier transform infrared (FTIR) microspectroscopy. Pollen germination in boron-deficient medium ranged from 18 to 24%, whereas pollen germination in standard medium reached 61%. Callose accumulated in the tip-regions of pollen tubes cultured in boron-deficient medium, but not in standard medium. Immunolabeling with antibody JIM5 revealed that acidic pectin preferentially accumulated in the tip regions of pollen tubes cultured in boron-deficient medium, whereas acidic pectin was weakly distributed along the entire lengths of pollen tubes cultured in standard medium. Esterified pectin, detected by immunolabeling with antibody JIM7, showed a similar distribution pattern in pollen tubes in both the boron-deficient and standard treatments. The FTIR spectra indicated slight increases in contents of phenolics and carboxylic acids and a substantial decrease in the content of saturated esters in boron-deficient pollen tubes compared with normal pollen tubes. The FTIR spectra confirmed that boron deficiency enhanced acidic pectin accumulation in pollen tubes, which may be associated with the increased content of carboxylic acid. We conclude that boron has a regulatory role in pollen germination and pollen tube growth.

Boron↗
PubMed2004

Basal defenses induced in pepper by lipopolysaccharides are suppressed by Xanthomonas campestris pv. vesicatoria.

The nonpathogenic hrcC mutant of Xanthomonas campestris pv. vesicatoria 85-10::hrpA22 multiplied in pepper leaves if it was mixed with pathogenic strains of X. campestris pv. vesicatoria. Reactions to the mutant alone included localized deposition of phenolics and callose in papillae, and alterations to the plant cell wall leading to increased electron density. Electron microscopy showed that the localized responses were suppressed in the presence of wild-type bacteria but other wall changes occurred at some sites, involving cellulose-rich ingrowth of the wall. Multiplication of the hrp mutant in mixed inocula was confirmed by tagging 85-10::hrpA22 using immunocytochemical location of AvrBs3 expressed from the plasmid pD36. Elicitors of callose deposition and other wall changes were isolated from the hrcC mutant. Activity in extracts of bacteria was attributed to the presence of high molecular weight lipopolysaccharides (LPS). Wild-type X. campestris pv. vesicatoria suppressed induction of structural changes caused by purified LPS. Results obtained suggest that effector proteins produced by phytopathogenic bacteria and delivered by the type III secretion system may have a key role in suppressing the basal defense responses activated by bacterial LPS, which lead to restricted multiplication of nonpathogens such as hrp mutants.

Bacterial Proteins↗
PubMed1992

Bicommissural neurones in the cerebral cortex of developing hamsters.

The trajectory, developmental time course, and origin of callosal fibres that recross through the anterior commissure were studied in developing hamsters, using carbocyanines in fixed brains on different ages. The bicommissural fibres were found in hamsters from E15 through P7, but disappeared after P7. By double labelling it was found that the neurones of origin of these bicommissural fibres were located in the lateral cortex within the region where the callosal zone of origin overlaps that of the anterior commissure. From these experiments, it was concluded that the axons of a group of cells in the lateral cortex of developing rodents are branched and grow transiently through both the callosum and the anterior commissure.

Animals↗
PubMed1999

VEP-interhemispheric transfer time in 20-32 Hz band in man.

This research aimed to investigate callosal transfer in the different frequency bands of VEP to lateralized reversal of checkerboard pattern as stimuli. The chosen band pass filters (4-8 Hz, 8-15 Hz, 15-20 Hz, 20-32 Hz) were applied to the VEPs of subjects, and four different components for each VEP were obtained. Latency differences between hemispheres for digitally unfiltered and filtered VEPs were computed to estimate IHTT. Different IHTTs in the theta (17 ms), alpha (9 ms) and beta1 (7 ms) bands from right to left and from left to right were estimated, supporting the previous report. Furthermore, a transfer within 4.5 ms from right to left in the 20-32 Hz band for the occipital lobe was found. In conclusion, our findings suggest that the frequency analysis of VEP to lateralized stimuli give us additional information, relating to the types of callosal fibers.

Adult↗
PubMed2000

An fMRI study of the lateralization of motor cortex activation in acallosal patients.

Transcranial magnetic stimulation (TMS) studies have suggested that callosal afferents may mediate inhibition of the ipsilateral motor cortex (IMC) during unilateral hand movements. To test this concept, we used fMRI to determine whether acallosal patients have increased IMC activation with either complex or simple unilateral finger movements. Neither the localization of motor cortical regions activated, the volumes of activation, or the relative hemispheric lateralization of activations were different between the patients and normal controls. The potential callosal inhibitory pathway identified by TMS therefore does not appear to contribute to the interhemispheric suppression of physiologically relevant activations in the motor cortex as measured by fMRI.

Adult↗
PubMedDate not supplied

Curvilinear and tubulonodular varieties of lipoma of the corpus callosum: an MR and CT study.

Until recently all callosal lipomas were grouped together. However, these lipomas can be clearly separated into two groups having different morphologies and associated brain anomalies. One group consists of anteriorly situated round or cylinder-shaped lipomas, termed tubulonodular. These lipomas are generally greater than 2 cm in diameter and have a high incidence of corpus callosum dysgenesis, frontal lobe anomalies, and frontal encephaloceles. The second group comprises thin, posteriorly situated lipomas curving around the splenium and are termed curvilinear. The curvilinear lipomas are generally associated with a normal corpus callosum and otherwise have a low incidence of associated anomalies. When anomalies do occur, they tend to be less severe than with the tubulonodular lipomas. We present four cases of curvilinear lipomas and review similar cases in the literature to explain why lipomas of the callosal region have such a variety of associated anomalies and morphologies. Our findings support the idea that both groups of lipomas develop in the region of the origin of the corpus callosum but form at different times during gestation, with the tubulonodular ones forming earlier and thus being associated with more severe cerebral anomalies.

Adult↗
PubMed1985

Topography of the human corpus callosum.

This study was undertaken to determine the topographical organization of fibers coursing through the human corpus callosum. We correlated the distribution of Wallerian degeneration in the corpus callosum with the anatomical sites of focal cortical lesions due to ischemic infarctions or circumscribed contusions. Fibers from the inferior frontal and anterior inferior parietal regions course through the rostrum and genu of the corpus callosum. Callosal connections from the temporo-parieto-occipital junctional region course through the splenium and caudal portion of the body of the corpus callosum. Both the superior parietal lobule and the occipital cortex give rise to interhemispheric fibers that course exclusively through the splenium of the corpus callosum. No callosal degeneration was associated with a cortical lesion in the anterior superior frontal region. The topographical organization of fibers in the human corpus callosum appears to be fairly similar to that found in the rhesus monkey (Macaca mulatta).

Animals↗
PubMed1996

The arteries of the corpus callosum: a microsurgical anatomic study.

OBJECTIVE: The corpus callosum is the major commissural pathway connecting the hemispheres of the human brain. It is particularly important, because various tumors and vascular lesions can be located in and around the corpus callosum, and it is a route through which pass several surgical approaches. Performing accurate surgery in this region and avoiding damage to normal structures require that the neurosurgeon have adequate knowledge of the anatomy of the intricate blood supply to this area. METHODS: In 20 cadaver brains, the arteries of the corpus callosum were examined under the operating microscope, with particular attention to the origin, course, anastomoses, number, and caliber of the arteries. RESULTS: In all specimens, the pericallosal and posterior pericallosal arteries were found to be the main sources of blood supply to the corpus callosum. In 80% of the specimens, the anterior communicating artery gave rise to either a subcallosal artery or a median callosal artery, each of which made a substantial contribution to the blood supply of the corpus callosum. A detailed examination of the anatomic features of all the main arteries of supply revealed anastomoses within the callosal sulcus that formed the pericallosal pial plexus. This network supplied the corpus callosum, the radiation of the corpus callosum, and the cingulate gyrus. CONCLUSION: Familiarity with the details of the vascularity of the corpus callosum is crucial when performing surgery in this region. The additional, significant data described expands the knowledge of this anatomy, which can enhance the surgeon's ability to accomplish a more accurate and successful exploration.

Cadaver↗
PubMed2004

Binding of arabinogalactan proteins by Yariv phenylglycoside triggers wound-like responses in Arabidopsis cell cultures.

Arabinogalactan-proteins (AGPs) are cell wall proteoglycans and are widely distributed in the plant kingdom. Classical AGPs and some nonclassical AGPs are predicted to have a glycosylphosphatidylinositol lipid anchor and have been suggested to be involved in cell-cell signaling. Yariv phenylglycoside is a synthetic probe that specifically binds to plant AGPs and has been used to study AGP functions. We treated Arabidopsis suspension cell cultures with Yariv phenylglycoside and observed decreased cell viability, increased cell wall apposition and cytoplasmic vesiculation, and induction of callose deposition. The induction of cell wall apposition and callose synthesis led us to hypothesize that Yariv binding of plant surface AGPs triggers wound-like responses. To study the effect of Yariv binding to plant surface AGPs and to further understand AGP functions, an Arabidopsis whole genome array was used to monitor the transcriptional modifications after Yariv treatment. By comparing the genes that are induced by Yariv treatment with genes whose expressions have been previously shown to be induced by other conditions, we conclude that the gene expression profile induced by Yariv phenylglycoside treatment is most similar to that of wound induction. It remains uncertain whether the Yariv phenylglycoside cross-linking of cell surface AGPs induces these genes through a specific AGP-based signaling mechanism or through a general mechanical perturbation of the cell surface.

Arabidopsis↗
PubMed2006

Arabidopsis transcriptome changes in response to phloem-feeding silverleaf whitefly nymphs. Similarities and distinctions in responses to aphids.

Phloem-feeding pests cause extensive crop damage throughout the world, yet little is understood about how plants perceive and defend themselves from these threats. The silverleaf whitefly (SLWF; Bemisia tabaci type B) is a good model for studying phloem-feeding insect-plant interactions, as SLWF nymphs cause little wounding and have a long, continuous interaction with the plant. Using the Affymetrix ATH1 GeneChip to monitor the Arabidopsis (Arabidopsis thaliana) transcriptome, 700 transcripts were found to be up-regulated and 556 down-regulated by SLWF nymphs. Closer examination of the regulation of secondary metabolite (glucosinolate) and defense pathway genes after SLWF-instar feeding shows that responses were qualitatively and quantitatively different from chewing insects and aphids. In addition to the RNA profile distinctions, analysis of SLWF performance on wild-type and phytoalexin-deficient4 (pad4) mutants suggests aphid and SLWF interactions with Arabidopsis were distinct. While pad4-1 mutants were more susceptible to aphids, SLWF development on pad4-1 and wild-type plants was similar. Furthermore, although jasmonic acid genes were repressed and salicylic acid-regulated genes were induced after SLWF feeding, cytological staining of SLWF-infested tissue showed that pathogen defenses, such as localized cell death and hydrogen peroxide accumulation, were not observed. Like aphid and fungal pathogens, callose synthase gene RNAs accumulated and callose deposition was observed in SLWF-infested tissue. These results provide a more comprehensive understanding of phloem-feeding insect-plant interactions and distinguish SLWF global responses.

Animals↗
PubMed2006

Vitamin B1-induced priming is dependent on hydrogen peroxide and the NPR1 gene in Arabidopsis.

Thiamine confers systemic acquired resistance (SAR) on susceptible plants through priming, leading to rapid counterattack against pathogen invasion and perturbation of disease progress. Priming reduces the metabolic cost required for constitutive expression of acquired resistance. To investigate the effects of priming by thiamine on defense-related responses, Arabidopsis (Arabidopsis thaliana) was treated with thiamine and effects of pathogen challenge on the production of active oxygen species, callose deposition, hypersensitive cell death, and pathogenesis-related 1 (PR1)/Phe ammonia-lyase 1 (PAL1) gene expression was analyzed. Thiamine did not induce cellular and molecular defense responses except for transient expression of PR1 per se; however, subsequent Pseudomonas syringae pv tomato challenge triggered pronounced cellular defense responses and advanced activation of PR1/PAL1 gene transcription. Thiamine treatment and subsequent pathogen invasion triggered hydrogen peroxide accumulation, callose induction, and PR1/PAL1 transcription activation in Arabidopsis mutants insensitive to jasmonic acid (jar1), ethylene (etr1), or abscisic acid (abi3-3), but not in plants expressing bacterial NahG and lacking regulation of SAR (npr1 [nonexpressor of PR genes 1]). Moreover, removal of hydrogen peroxide by catalase almost completely nullified cellular and molecular defense responses as well as SAR abolishing bacterial propagation within plants. Our results indicated that priming is an important cellular mechanism in SAR by thiamine and requires hydrogen peroxide and intact NPR1.

Arabidopsis↗
PubMed1997

Al-Induced, 51-Kilodalton, Membrane-Bound Proteins Are Associated with Resistance to Al in a Segregating Population of Wheat.

Incorporation of 35S into protein is reduced by exposure to Al in wheat (Triticum aestivum), but the effects are genotype-specific. Exposure to 10 to 75 [mu]M Al had little effect on 35S incorporation into total protein, nuclear and mitochondrial protein, microsomal protein, and cytosolic protein in the Al-resistant cultivar PT741. In contrast, 10 [mu]M Al reduced incorporation by 21 to 38% in the Al-sensitive cultivar Katepwa, with effects becoming more pronounced (31-62%) as concentrations of Al increased. We previously reported that a pair of 51-kD membrane-bound proteins accumulated in root tips of PT741 under conditions of Al stress. We now report that the 51-kD band is labeled with 35S after 24 h of exposure to 75 [mu]M Al. The specific induction of the 51-kD band in PT741 suggested a potential role of one or both of these proteins in mediating resistance to Al. Therefore, we analyzed their expression in single plants from an F2 population arising from a cross between the PT741 and Katepwa cultivars. Accumulation of 1,3-[beta]-glucans (callose) in root tips after 24 h of exposure to 100 [mu]M Al indicated that this population segregated for Al resistance in about a 3:1 ratio. A close correlation between resistance to Al (low callose content of root tips) and accumulation of the 51-kD band was observed, indicating that at least one of these proteins cosegregates with the Al-resistance phenotype. As a first step in identifying a possible function, we have demonstrated that the 51-kD band is most clearly associated with the tonoplast. Whereas Al has been reported to stimulate the activity of the tonoplast H+-ATPase and H+-PPase, antibodies raised against these proteins did not cross-react with the 51-kD band. Efforts are now under way to purify this protein from tonoplast-enriched fractions.

Journal Article↗
PubMed2001

Pollen tubes of Nicotiana alata express two genes from different beta-glucan synthase families.

The walls deposited by growing pollen tubes contain two types of beta-glucan, the (1,3)-beta-glucan callose and the (1,4)-beta-glucan cellulose, as well as various alpha-linked pectic polysaccharides. Pollen tubes of Nicotiana alata Link et Otto, an ornamental tobacco, were therefore used to identify genes potentially encoding catalytic subunits of the callose synthase and cellulose synthase enzymes. Reverse transcriptase-polymerase chain reactions (RT-PCR) with pollen-tube RNA and primers designed to conserved regions of bacterial and plant cellulose synthase (CesA) genes amplified a fragment that corresponded to an abundantly expressed cellulose-synthase-like gene named NaCslD1. A fragment from a true CesA gene (NaCesA1) was also amplified, but corresponding cDNAs could not be identified in a pollen-tube library, consistent with the very low level of expression of the NaCesA1 gene. RT-PCR with pollen-tube RNA and primers designed to regions conserved between the fungal FKS genes [that encode (1,3)-beta-glucan synthases] and their presumed plant homologs (the Gsl or glucan-synthase-like genes) amplified a fragment that corresponded to an abundantly expressed gene named NaGsl1. A second Gsl gene detected by RT-PCR (NaGsl2) was expressed at low levels in immature floral organs. The structure of full-length cDNAs of NaCslD1, NaCesA1, and NaGsl1 are presented. Both NaCslD1 and NaGsl1 are predominantly expressed in the male gametophyte (developing and mature pollen and growing pollen tubes), and we propose that they encode the catalytic subunits of two beta-glucan synthases involved in pollen-tube wall synthesis. Different beta-glucans deposited in one cell type may therefore be synthesized by enzymes from different gene families.

Amino Acid Sequence↗
PubMed2001

Class III pistil-specific extensin-like proteins from tobacco have characteristics of arabinogalactan proteins.

Class III pistil-specific extensin-like proteins (PELPIII) are specifically localized in the intercellular matrix of tobacco (Nicotiana tabacum) styles. After pollination the majority of PELPIII are translocated into the callosic layer and the callose plugs of the pollen tubes, which could suggest a function of PELPIII in pollen tube growth. PELPIII may represent one of the chemical and/or physical factors from the female sporophytic tissue that contributes to the difference between in vivo and in vitro pollen tube growth. PELPIII glycoproteins were purified and biochemically characterized. Because of their high proline (Pro) and hydroxy-Pro (Hyp) content, PELPIII proteins belong to the class of Pro/Hyp-rich glycoproteins. The carbohydrate moiety of PELPIII is attached through O-glycosidic linkages and comprises more than one-half the total glycoprotein. Deglycosylation of PELPIII revealed two backbones, both reacting with PELPIII-specific antibodies. N-terminal amino acid sequencing of these backbones showed that PELPIII is encoded by the MG14 and MG15 genes. Two heterogeneous N-terminal sequences of MG14 and MG15, both starting downstream of the predicted signal peptide cleavage site, seem to be present, which indicates a novel N-terminal processing. Monosaccharide analysis showed that the carbohydrate moiety of PELPIII almost completely consists of arabinose and galactose in an equal molar ratio. Carbohydrate linkage analysis showed terminal and 2-linked arabinofuranosyl residues, as well as terminal and 6-, 3-, and 3,6-linked galactopyranosyl residues to be present, indicating the presence of both extensin-like and Type II arabinogalactan oligosaccharide units. The ability of beta-glucosyl Yariv reagent to bind with PELPIII confirmed the arabinogalactan protein-like characteristics of these proteins.

Amino Acid Sequence↗