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1.
Vega-Maray AM Fernández-González D Valencia-Barrera R Suárez-Cervera M 《Protoplasma》2006,228(1-3):115-120
Summary. Rapid diffusion of allergenic proteins in isotonic media has been demonstrated for different pollen grains. Upon contact with
stigmatic secretion or with the mucosa of sensitive individuals, pollen grains absorb water and release soluble low-molecular-weight
proteins, these proteins enter in the secretory pathway in order to arrive at the cell surface. In this study we located allergenic
proteins in mature and hydrated-activated pollen grains of Parietaria judaica L. (Urticaceae) and studied the diffusion of these proteins during the first 20 min of the hydration and activation processes.
A combination of transmission electron microscopy and immunocytochemical methods was used to locate these proteins in mature
pollen and in pollen grains after different periods of hydration and activation processes. Activated proteins reacting with
antibodies in human serum from allergic patients were found in the cytoplasm, wall, and exudates from the pollen grains. The
allergenic component of these pollen grains changes according to the pollen state; the presence of these proteins in the exine,
the cytoplasm, and especially in the intine and in the material exuded from the pollen grains, is significant in the hydrated-activated
studied times, whereas this presence is not significant in mature pollen grains. The rapid activation and release of allergenic
proteins of P. judaica pollen appears to be the main cause of the allergenic activity of these pollen grains.
Correspondence and reprints: Department of Plant Biology, Faculty of Biology, University of León, Campus de Vegazana, 24071
León, Spain. 相似文献
2.
Yi -Qin Li Claudia Faleri Anja Geitmann Hong -Qi Zhang Mauro Cresti 《Protoplasma》1995,189(1-2):26-36
Summary The monoclonal antibodies JIM 5 (against unesterified pectin), JIM 7 (against methyl esterified pectin), MAC 207 (against arabinogalactan proteins, AGPs), and JIM 8 (against a subset of AGPs) were utilized singly or in combinations for immunogold labelling of germinated pollen grains and pollen tubes ofNicotiana tabacum. Pectins were localized in the inline of pollen grain, unesterified pectin being more abundant than the esterified one. AGPs were co-localized with pectin in the inline, but were present preferably close to the plasma membrane. In pollen tubes, AGPs, unesterified and esterified pectins were co-localized in the outer and middle layers of the cell wall. The density of the epitopes was not uniform along the length of the pollen tube, but showed alterations. In the pollen tube tip wall esterified pectin was abundantly present, but not AGPs. In the cytoplasm esterified pectin and AGPs were detected in Golgi derived vesicles, indicating their role in the pathway of the cell wall precursors. In the cell wall of generative cell only AGPs, but no pectins were localized. The co-localization of pectins and AGPs in the cell wall of pollen grain and pollen tube might play an important role, not only in maintenance of the cell shape, but also in cell-cell interaction during pollen tube growth and development.Abbreviations AGP
arabinogalactan protein
- BSA
bovine serum albumin
- GA
glutaraldehyde
- MAb
monoclonal antibody
- NGS
normal goat serum
- PFA
paraformaldehyde 相似文献
3.
4.
MarÍa Suárez‐Cervera Ana Vega‐Maray Teresa Castells F. Javier RodrÍguez‐Rajo Juan A. Asturias Annick Le Thomas 《Grana》2013,52(4):272-284
The aim of the present study is to localise non‐specific lipid transfer proteins (nsLTPs), obtained from Rosaceae (Prunus persica (L.) Batsch) in pollen grains of taxonomically distant plants, such as Iridaceae (Aristea latifolia G.J. Lewis), Platanaceae (Platanus acerifolia (Aiton) Willd.) and Urticaceae (Parietaria judaica L.), in order to compare pollen and nsLTP diversity. A combination of transmission electron microscopy, immunocytochemical techniques, and rabbit specific antiserum against peach nsLTPs (Pru p 3) were used. Abundant labelling to Pru p 3‐like proteins was observed in the cytoplasm, walls and pollenkitt of A. latifolia pollen grains. The presence of nsLTPs associated with the pollenkitt proves that it takes part in the defence mechanism of pollen grains. The labelling was less intense in the cytoplasm and walls of P. acerifolia. Immuno‐stained gold particles were associated with the vacuoles, lipid inclusions, endoplasmic reticulum and Golgi complex. No significant labelling was found in the P. judaica pollen grains incubated with anti‐Pru p 3 polyclonal antibodies. These results indicate important variations in the nsLTPs of different pollen species. Consequently, no taxonomic relationship between pollen grains and nsLTPs could be established. 相似文献
5.
Abstract Pollen grain viability and starch presence in pollen were followed during the long blooming period (May-November) of Parietaria judaica, the most widespread pellitory in Italy, responsible for many cases of allergic diseases. Observations were carried out near Siena (Tuscany), in the years 1978 and 1979. Viability is high in late spring and early autumn, and pollen grains are starchy. The presence of starchless grains is always related to low viability: a production of pollen with a low viability occurs at the beginning and end of blooming, and also in summer, during drought periods. Pollen grain viability varies widely during the blooming period, and its variations are correlated to meteorological conditions, mainly rainfall and temperature. Allergic diseases due to pellitory, however, are mainly reported between April and July, and on the other hand a small amount of patients states that they suffer all the year round. The discrepancies between the periods of viable pollen production and of declared symptoms by patients, are discussed. 相似文献
6.
Libin Chen Chonghui Ji Degui Zhou Xin Gou Jianian Tang Yongjie Jiang Jingluan Han Yao-Guang Liu Letian Chen Yongyao Xie 《遗传学报》2022,49(5):481-491
In plants, lipid transfer proteins(LTPs) transport pollen wall constituents from the tapetum to the exine, a process essential for pollen wall development. However, the functional cooperation of different LTPs in pollen wall development is not well understood. In this study, we have identified and characterized a grassspecific LTP gene, Os LTP47, an important regulator of pollen wall formation in rice(Oryza sativa). Os LTP47 encodes a membrane-localized LTP and in vitro lipid-binding assays conf... 相似文献
7.
Mario Lachapelle Moise Bendayan 《Virchows Archiv. B, Cell pathology including molecular pathology》1991,60(1):105-111
Actin and alpha-actinin immunoreactive sites have been localized at the electron microscope level by the protein A-gold immunocytochemical
technique in podocytes of normal and nephrotic rat renal tissues. In normal renal glomeruli, fibrillar networks located in
the core of foot processes or bundles of micro filaments interconnecting them were found to be labelled for these two cytoskeletal
proteins. On the other hand, in nephrotic renal glomeruli, concomitant with the loss of podocytic foot processes a reorganization
of the podocytic cytoskeleton and a concentration of some of its elements into thick uniform bands was observed. Actin and
alpha-actinin were revealed in these bands. Control experiments confirmed the specificity of the labelling obtained. Our results
suggest that normal podocytes contain an actin-based contractile system that might contribute to the maintenance of the particular
cell shape of these cells and that the rearrangement of the podocytic cyto-skeleton occurring in the nephrotic syndrome might
account for the changes in the foot processes and contribute to the alteration in glomerular function.
This work was supported by grants from the Medical Research Council of Canada 相似文献
8.
9.
Li Yi-qin T. H. Tsao 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,71(2):263-267
Summary A method was worked out using trifluoromethanesulfonic acid (TFMS) as a reagent to split the covalently bound proteins, which are NaCl insoluble, from pollen tube walls of Lilium longiflorum, leaving the peptide bonds essentially intact. After electrophoretic separation, comparisons were made among these proteins from pollen grains and pollen tubes grown in vitro and in styles after self- and cross-pollination. It was found that a) the patterns of covalently bound wall proteins were different between tubes grown in vitro and in vivo; b) fewer bands were found in covalently bound wall proteins than that in noncovalently bound proteins; c) the bands remained almost the same no matter whether the tubes had been cross pollinated or self pollinated, indicating that while the noncovalently bound proteins were involved in incompatibility as shown in the previous paper, the covalently bound proteins may only serve as a structural component, having little to do with incompatibility. 相似文献
10.
11.
The distribution of cellulose and callose in the walls of pollen tubes and grains of Nicotiana tabacum L. was examined by electron microscopy using gold-labelled cellobiohydrolase for cellulose and a (1,3)-β-D-glucan-specific
monoclonal antibody for callose. These probes provided the first direct evidence that cellulose co-locates with callose in
the inner, electron-lucent layer of the pollen-tube wall, while both polymers are absent from the outer, fibrillar layer.
Neither cellulose nor callose are present in the wall at the pollen-tube tip or in cytoplasmic vesicles. Cellulose is first
detected approximately 5–15 μm behind the growing tube tip, just before a visible inner wall layer commences, whereas callose
is first observed in the inner wall layer approximately 30 μm behind the tip. Callose was present throughout transverse plugs,
whereas cellulose was most abundant towards the outer regions of these plugs. This same distribution of cellulose and callose
was also observed in pollen-tube walls of N. alata Link et Otto, Brassica campestris L. and Lilium longiflorum Thunb. In pollen grains of N. tabacum, cellulose is present in the intine layer of the wall throughout germination, but no callose is present. Callose appears
in grains by 4 h after germination, increasing in amount over at least the first 18 h, and is located at the interface between
the intine and the plasma membrane. This differential distribution of cellulose and callose in both pollen tubes and grains
has implications for the nature of the β-glucan biosynthetic machinery.
Received: 20 February 1988 / Accepted: 25 March 1998 相似文献
12.
13.
The numbers of embryogenic (S) grains present in in-situ mature anthers of Nicotiana tabacum L. were compared to the numbers of embryos and plantlets produced in cultured anthers excised at the optimal mitotic stage of development for anther culture. The Feulgen technique of staining embryos caused a considerable loss of grains from cultured anthers but this did not seriously affect the determination of the percentage of embryos present. In no instance did the numbers of embryos produced exceed the maximum number of S grains found, and the distributions of S grain and embryo frequencies in anthers were similar. In rare instances S grains which had undergone the first embryogenic division were observed in situ. The results indicate that all grains capable of embryogenesis are determined during early flower formation and that their number is not increased by in vitro culture. 相似文献
14.
Membrane contact sites, where two organelles are in close proximity, are critical regulators of cellular membrane homeostasis, with roles in signaling, lipid metabolism, and ion dynamics. A growing catalog of specialized lipid transfer proteins carry out lipid exchange at these sites. Currently characterized eukaryotic lipid transport proteins are shuttles that typically extract a single lipid from the membrane of the donor organelle, solubilize it during transport through the cytosol, and deposit it in the acceptor organelle membrane. Here, we highlight the recently identified chorein_N family of lipid transporters, including the Vps13 proteins and the autophagy protein Atg2. These are elongated proteins that, distinct from previously characterized transport proteins, bind tens of lipids at once. They feature an extended channel, most likely lined with hydrophobic residues. We discuss the possibility that they are not shuttles but instead are bridges between membranes, with lipids traversing the cytosol via the hydrophobic channel. 相似文献
15.
María Suárez-Cervera Juan A. Asturias Ana Vega-Maray Teresa Castells Carmen López-Iglesias Ignacio Ibarrola M. Carmen. Arilla Nina Gabarayeva Juan A. Seoane-Camba 《Sexual plant reproduction》2005,18(3):101-112
Platanus acerifolia (Aiton) Willdenow is a plane tree, widely grown as an ornamental tree in many cities of the United States and Western Europe,
which has become an important source of airborne allergens in our cities. The aim of the present study is to immunolocalize
the major allergens in the pollen grain and to examine their potential function in the fertilization process. Observations
were made in mature and hydrated, activated pollen of P. acerifolia for 5, 15, 30 min and 2 h in the germination medium. Specimens were fixed using freezing protocols for transmission electron
microscopy (TEM). For immunogold labelling, cryosections and resin-embedded ultrathin sections were incubated using rabbit
antisera against the purified pollen allergens Pla a 1 and Pla a 2. Elution of P. acerifolia allergens took place after 5 min of pollen incubation in buffered medium. Intense labelling of Pla a 1 and Pla a 2 was detected
after pollen exudates were released. In pollen grains, Pla a 1 was predominantly localized in concentric cisternae of the
endoplasmic reticulum (ER), situated between the vegetative nucleus and the generative cell, and was released from pollen
grains 5 min after hydration; cytoplasmic localization decreased 15 min after hydration. In pollen grains, glycoprotein Pla
a 2 was abundant in association with Golgi cisternae and vesicles situated in the apertural periphery of the mature pollen
grains. Pla a 2 proteins were also detected in ER and in the generative cell wall. Immunolabelling of Pla a 2 decreased 5 min
after pollen hydration but was again intense after 15–30 min in germination medium, presumably as a consequence of renewed
expression and glycosylation of this protein. Pla a 1 belongs to a new class of allergens related to proteinaceous invertase
and pectin methyl esterase inhibitors (PII, PMEI) which could be involved in membrane protection and pectin de-esterification
control during pollen hydration. Pla a 2 has an exopolygalacturonase (PG) enzymatic activity consistent with pollen-stigma
adhesion mechanisms or compatibility systems. Moreover, the expression of Pla a 2 found 15–30 min after hydration might contribute
to pollen-tube growth and the modification of transmitting tissue cell walls. The abundant production and elution of Pla a
1 and Pla a 2 proteins may alter the environment in which pollen tube elongation occurs, thus promoting a potential crosstalk
between the pollen and the gynoecium. 相似文献
16.
Lipid transfer protein genes specifically expressed in barley leaves and coleoptiles 总被引:5,自引:0,他引:5
K. Gausing 《Planta》1994,192(4):574-580
Genes/cDNAs encoding so-called lipid-transfer proteins (LTPs) have been isolated from a variety of tissues from different plants, but the in-vivo function of the LTP proteins is not yet known. In barley (Hordeum vulgare L.), the LTP1 gene (encoding a probable amylase/ protease inhibitor, Mundy and Rogers 1986, Planta 169, 51–63) is active in aleurone tissue, and in this paper two LTP-encoding cDNAs isolated from green leaves are described. The encoded proteins start with signal sequences, they are 75% homologous to each other, 60–63% homologous to rice aleurone LTP and maize seed/ coleoptile LTP, but only 48% homologous to barley aleurone LTP. Northern hybridization experiments established that the two seedling-specific genes are both highly expressed in leaves and coleoptiles whereas the LTP1 gene is inactive in seedlings. No LTP gene expression was detected in roots using either seedling or aleurone cDNA clones as probes. Tissue-print hybridization indicates that the LTP genes are first expressed in young epidermal cells in leaves and coleoptiles, and subsequently expressed in the vascular strands. Genomic Southern analysis indicates that the barley LTP gene family has four to six members.Abbreviation LTP
lipid transfer protein
I thank Dr. J. Mundy, Carlsberg Research Laboratory, Copenhagen, Denmark for the PAPI cDNA clone and R. Barkardottir, Department of Molceular Biology, University of Aarhus, Denmark for providing RNA for some of the Northern analyses. I also thank I. Bjørndal and L. Kjeldbjerg for excellent technical assistance. This work was supported by the The Danish Biotechnology Programme. 相似文献
17.
Philip E. Taylor Kerstin Spuck Penelope M. Smith Jenneth M. Sasse Takao Yokota Peter G. Griffiths Donald W. Cameron 《Planta》1993,189(1):91-100
Bioactive brassinosteroids have been localized in developing and mature pollen of anhydrously fixed rye-grass (Lolium perenne) by immunocytochemistry using polyclonal antibodies to castasterone generated in rabbits. Tricellular pollen fixed by freeze-substitution was also labelled in the starch granules. Study of the developmental sequence of the pollen through the microsporocyte, microspore, bicellular and tricellular stages showed that the brassinosteroids were increasingly sequestered in starch granules as the amyloplasts matured, supporting the view that these are storage organelles for these potent plant growth promoters. In bicellular pollen, heavy labelling was seen in the zone within 0.5 m of the starch granule, where stromal tissue remains. Thus, the stroma may be the site of synthesis of these compounds. During aqueous fixation, the brassinosteroids leached from the starch granules of tricellular pollen, indicating that they would be quickly available after imbibition to influence the physiology of germinating pollen. The results from high-performance liquid chromatography of dansylaminophenylboronates from partially purified extracts of freshly dehisced tricellular pollen of rye-grass showed 25-methylcastasterone may be a minor component, together with two unknown peaks. No specific binding of brassinolide to any soluble proteins extracted from tricellular rye-grass pollen was observed using the antibodies in gel electrophoresis or enzyme-linked immunosorbent assays.Abbreviations HPLC
high-performance liquid chromatography
- Rt
retention time
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
We thank the Australian Research Council for support, the N.S.W. Department of Agriculture for rice seed, Professor K. Mori and the Zen-Noh Corporation for authentic brassinosteroid samples, Dr. I. Hudson for statistical advice, Dr. A. Bacic and Ms. I. Bonig for helpful discussion, and J.M.S. thanks Professor R.B. Knox for laboratory facilities. 相似文献
18.
Ultrastructural immunocytochemical localization of LH and FSH in the pituitary of the untreated male rat 总被引:1,自引:0,他引:1
Summary Rapid freeze-substitution fixation was employed in immunocytochemical studies on the localization of LH and FSH in the typical gonadotrophs of the anterior pituitary in the untreated male rat; a modification of a recently described ferritin antibody method (Inoue et al. 1982) was used in these studies. It was shown that rapid freeze-substitution fixation provides good preservation not only of the ultrastructure but also of the antigenicity. Both LH and FSH were clearly demonstrated in the same gonadotrophic cells, but the subcellular localization of these gonadotrophins differed: (i) LH was mainly located in small secretory granules, 250–300 nm in diameter; (ii) FSH was mainly present in large secretory granules, up to 500 nm in diameter. In the pituitary gland of the adult male rat, all gonadotrophs that react to antibodies against gonadotrophins are characterized by small and large secretory granules. Other types of cells of the anterior pituitary containing either small secretory granules or resembling corticotrophs with secretory granules assembled at cell periphery did not react to either anti-LH beta or anti-FSH beta serum.For light microscopy, the peroxidase antibody method was used. All of the gonadotrophin-positive cells contain both LH and FSH. None of the pituitary cells reacted to antibody against only one gonadotrophin. However, some cells are LH-rich while other cells are FSH-rich. 相似文献
19.
The monoclonal antibody OL-1 and transmission electron microscopy were used to locate immunologically the major allergen of olive pollen. Ole e I, during pollen grain development. Within the pollen grain, allergenic proteins are located in the cisternae of the rough endoplasmic reticulum. Our findings indicate that the synthesis of these proteins starts in the vegetative cytoplasm of olive pollen during the early maturation stage.Dedicated to Professor Andreas Sievers on the occasion of his retirement 相似文献
20.
Ultrastructural localization of prolactin,growth hormone and luteinizing hormone by immunocytochemical techniques in the bovine pituitary 总被引:2,自引:0,他引:2
Summary The technique of ultrastructural immunocytochemistry involving the unlabeled antibody and the soluble peroxidase-antiperoxidase complex was used to identify and describe the prolactin (P) cells, somatotropic (STH) cells and luteinizing hormone (LH) cells in the bovine anterior pituitary gland. This method was used to localize the three hormones at the electron microscopic level. Staining of varying intensity was found on the secretory granules and on the small granules and vesicles within the Golgi complex. No stain was found in nuclei, on mitochondria or in the endoplasmic reticulum. 相似文献