scholarly journals Comparative analyses of angiosperm secretomes identify apoplastic pollen tube functions and novel secreted peptides

2020 ◽  
Author(s):  
María Flores-Tornero ◽  
Lele Wang ◽  
David Potěšil ◽  
Said Hafidh ◽  
Frank Vogler ◽  
...  
Keyword(s):  
Cell Wall ◽  
Pollen Tube ◽  
Turgor Pressure ◽  
Pollen Grains ◽  
Pollen Tubes ◽  
Secreted Proteins ◽  
Reproductive Tissues ◽  
Small Proteins ◽  
Secreted Peptides

Abstract Key message Analyses of secretomes of in vitro grown pollen tubes from Amborella, maize and tobacco identified many components of processes associated with the cell wall, signaling and metabolism as well as novel small secreted peptides. Abstract Flowering plants (angiosperms) generate pollen grains that germinate on the stigma and produce tubes to transport their sperm cells cargo deep into the maternal reproductive tissues toward the ovules for a double fertilization process. During their journey, pollen tubes secrete many proteins (secreted proteome or secretome) required, for example, for communication with the maternal reproductive tissues, to build a solid own cell wall that withstands their high turgor pressure while softening simultaneously maternal cell wall tissue. The composition and species specificity or family specificity of the pollen tube secretome is poorly understood. Here, we provide a suitable method to obtain the pollen tube secretome from in vitro grown pollen tubes of the basal angiosperm Amborella trichopoda (Amborella) and the Poaceae model maize. The previously published secretome of tobacco pollen tubes was used as an example of eudicotyledonous plants in this comparative study. The secretome of the three species is each strongly different compared to the respective protein composition of pollen grains and tubes. In Amborella and maize, about 40% proteins are secreted by the conventional “classic” pathway and 30% by unconventional pathways. The latter pathway is expanded in tobacco. Proteins enriched in the secretome are especially involved in functions associated with the cell wall, cell surface, energy and lipid metabolism, proteolysis and redox processes. Expansins, pectin methylesterase inhibitors and RALFs are enriched in maize, while tobacco secretes many proteins involved, for example, in proteolysis and signaling. While the majority of proteins detected in the secretome occur also in pollen grains and pollen tubes, and correlate in the number of mapped peptides with relative gene expression levels, some novel secreted small proteins were identified. Moreover, the identification of secreted proteins containing pro-peptides indicates that these are processed in the apoplast. In conclusion, we provide a proteome resource from three distinct angiosperm clades that can be utilized among others to study the localization, abundance and processing of known secreted proteins and help to identify novel pollen tube secreted proteins for functional studies.

scholarly journals Plant AP180 N-Terminal Homolog Proteins Are Involved in Clathrin-Dependent Endocytosis during Pollen Tube Growth in Arabidopsis thaliana

2019 ◽  
Vol 60 (6) ◽  
pp. 1316-1330 ◽  
Author(s):  
Minako Kaneda ◽  
Chlo� van Oostende-Triplet ◽  
Youssef Chebli ◽  
Christa Testerink ◽  
Sebastian Y Bednarek ◽  
...  
Keyword(s):  
Cell Wall ◽  
Pollen Tube ◽  
Membrane Trafficking ◽  
Pollen Tubes ◽  
Wall Material ◽  
Knockout Mutant ◽  
Cell Wall Assembly ◽  
Morphological Defects ◽  
Fluorescent Tagging

Abstract Polarized cell growth in plants is maintained under the strict control and exquisitely choreographed balance of exocytic and endocytic membrane trafficking. The pollen tube has become a model system for rapid polar growth in which delivery of cell wall material and membrane recycling are controlled by membrane trafficking. Endocytosis plays an important role that is poorly understood. The plant AP180 N-Terminal Homolog (ANTH) proteins are putative homologs of Epsin 1 that recruits clathrin to phosphatidylinositol 4, 5-bisphosphate (PIP2) containing membranes to facilitate vesicle budding during endocytosis. Two Arabidopsis ANTH encoded by the genes AtAP180 and AtECA2 are highly expressed in pollen tubes. Pollen tubes from T-DNA inserted knockout mutant lines display significant morphological defects and unique pectin deposition. Fluorescent tagging reveals organization into dynamic foci located at the lateral flanks of the pollen tube. This precisely defined subapical domain coincides which clathrin-mediated endocytosis (CME) and PIP2 localization. Using a liposome-protein binding test, we showed that AtECA2 protein and ANTH domain recombinant proteins have strong affinity to PIP2 and phosphatidic acid containing liposomes in vitro. Taken together these data suggest that Arabidopsis ANTH proteins may play an important role in CME, proper cell wall assembly and morphogenesis.

scholarly journals Pollen tube growth of Paeonia tenuifolia L. (Paeoniaceae) in vitro and in vivo

1970 ◽  
Vol 40 (1) ◽  
pp. 93-95 ◽  
Author(s):  
Feruzan Dane ◽  
Nuran Ekici
Keyword(s):  
Pollen Tube ◽  
Pollen Germination ◽  
Pollen Viability ◽  
Pollen Grains ◽  
Pollen Tubes ◽  
In Vivo Studies ◽  
In Vivo Growth ◽  
Paeonia Tenuifolia

In vitro and in vivo studies on pollen germination of Paeonia tenuifolia L. (Paeoniaceae) revealed that pollen grains are shed at two-celled stage. Normal and abnormal pollens were observed. Pollen viability was recorded between 55 and 75%. In vitro studies revealed 85% germination and usually the germination was monosphonic. Some pollen tubes with swollen tube tip and undulations were found. Styles and stigma were found to contain many pollen tubes 24 hrs after pollination. Key words: Paeonia tenuifolia; Pollen tube; In vitro growth; In vivo growth  DOI: http://dx.doi.org/10.3329/bjb.v40i1.8003 Bangladesh J. Bot. 40(1): 93-95, 2011 (June)

scholarly journals Structure of the generative cell wall complex after freeze substitution in pollen tubes of Nicotiana and Impatiens

1987 ◽  
Vol 88 (3) ◽  
pp. 373-378
Author(s):  
M. CRESTI ◽  
S. A. LANCELLE ◽  
P. K. HEPLER
Keyword(s):  
Cell Wall ◽  
Plasma Membranes ◽  
Generative Cell ◽  
Pollen Grains ◽  
Freeze Substitution ◽  
Pollen Tubes ◽  
Wall Material ◽  
Cell Cytoplasm ◽  
Stage Of Development

The mature generative cell in pollen grains and pollen tubes is surrounded by a wall complex that includes two plasma membranes, one facing the generative cell cytoplasm and one facing the vegetative cell cytoplasm, and usually some intervening wall material. After conventional chemical fixation, the two plasma membranes are very uneven and often appear to be joined, giving the impression that numerous plasmodesmata connect the vegetative and generative cells. These areas alternate with swollen, distorted areas, which give the wall complex the appearance of being composed of a chain of vesicles. Utilizing rapid freeze fixation and freeze substitution, we have re-examined the ultrastructure of the generative cell wall complex from pollen tubes grown in vitro, and the differences are striking. The two plasma membranes are very smooth and closely appressed to a layer of wall material. Occasionally the wall complex contains swollen areas, or varicosities, and these may contain pockets of lightly stained material, but again the surrounding plasma membranes are tightly appressed to these areas. Plasmodesmata are not seen, but this does not eliminate the possibility that they may exist at an earlier stage of development.

scholarly journals Protein Analysis of Pollen Tubes after the Treatments of Membrane Trafficking Inhibitors Gains Insights on Molecular Mechanism Underlying Pollen Tube Polar Growth

2021 ◽  
Vol 40 (2) ◽  
pp. 205-222
Author(s):  
Monica Scali ◽  
Alessandra Moscatelli ◽  
Luca Bini ◽  
Elisabetta Onelli ◽  
Rita Vignani ◽  
...  
Keyword(s):  
Pollen Tube ◽  
Actin Cytoskeleton ◽  
Membrane Trafficking ◽  
Tip Growth ◽  
Male Gametophyte ◽  
Pollen Tubes ◽  
Structural Features ◽  
Two Dimensional Gel Electrophoresis ◽  
Depth Analysis

AbstractPollen tube elongation is characterized by a highly-polarized tip growth process dependent on an efficient vesicular transport system and largely mobilized by actin cytoskeleton. Pollen tubes are an ideal model system to study exocytosis, endocytosis, membrane recycling, and signaling network coordinating cellular processes, structural organization and vesicular trafficking activities required for tip growth. Proteomic analysis was applied to identifyNicotiana tabacumDifferentially Abundant Proteins (DAPs) after in vitro pollen tube treatment with membrane trafficking inhibitors Brefeldin A, Ikarugamycin and Wortmannin. Among roughly 360 proteins separated in two-dimensional gel electrophoresis, a total of 40 spots visibly changing between treated and control samples were identified by MALDI-TOF MS and LC–ESI–MS/MS analysis. The identified proteins were classified according to biological processes, and most proteins were related to pollen tube energy metabolism, including ammino acid synthesis and lipid metabolism, structural features of pollen tube growth as well modification and actin cytoskeleton organization, stress response, and protein degradation. In-depth analysis of proteins corresponding to energy-related pathways revealed the male gametophyte to be a reliable model of energy reservoir and dynamics.

scholarly journals Hyperoside promotes pollen tube growth by regulating the depolymerization effect of actin-depolymerizing factor 1 on microfilaments in okra

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Biying Dong ◽  
Qing Yang ◽  
Zhihua Song ◽  
Lili Niu ◽  
Hongyan Cao ◽  
...  
Keyword(s):  
Pollen Tube ◽  
Pollen Tube Growth ◽  
Pollen Germination ◽  
Pollen Tubes ◽  
Tube Growth ◽  
Actin Depolymerizing Factor ◽  
Specific Mechanism ◽  
Promoting Effect ◽  
New Research

AbstractMature pollen germinates rapidly on the stigma, extending its pollen tube to deliver sperm cells to the ovule for fertilization. The success of this process is an important factor that limits output. The flavonoid content increased significantly during pollen germination and pollen tube growth, which suggests it may play an important role in these processes. However, the specific mechanism of this involvement has been little researched. Our previous research found that hyperoside can prolong the flowering period of Abelmoschus esculentus (okra), but its specific mechanism is still unclear. Therefore, in this study, we focused on the effect of hyperoside in regulating the actin-depolymerizing factor (ADF), which further affects the germination and growth of pollen. We found that hyperoside can prolong the effective pollination period of okra by 2–3-fold and promote the growth of pollen tubes in the style. Then, we used Nicotiana benthamiana cells as a research system and found that hyperoside accelerates the depolymerization of intercellular microfilaments. Hyperoside can promote pollen germination and pollen tube elongation in vitro. Moreover, AeADF1 was identified out of all AeADF genes as being highly expressed in pollen tubes in response to hyperoside. In addition, hyperoside promoted AeADF1-mediated microfilament dissipation according to microfilament severing experiments in vitro. In the pollen tube, the gene expression of AeADF1 was reduced to 1/5 by oligonucleotide transfection. The decrease in the expression level of AeADF1 partially reduced the promoting effect of hyperoside on pollen germination and pollen tube growth. This research provides new research directions for flavonoids in reproductive development.

scholarly journals Prunus necrotic ringspot virus Early Invasion and Its Effects on Apricot Pollen Grain Performance

2007 ◽  
Vol 97 (8) ◽  
pp. 892-899 ◽  
Author(s):  
Khalid Amari ◽  
Lorenzo Burgos ◽  
Vicente Pallas ◽  
María Amelia Sanchez-Pina
Keyword(s):  
Developmental Stages ◽  
Generative Cell ◽  
Pollen Grains ◽  
Growth Zone ◽  
Pollen Tubes ◽  
Climatic Conditions ◽  
Pollen Grain ◽  
Ringspot Virus ◽  
Prunus Necrotic Ringspot Virus

The route of infection and the pattern of distribution of Prunus necrotic ringspot virus (PNRSV) in apricot pollen were studied. PNRSV was detected both within and on the surface of infected pollen grains. The virus invaded pollen during its early developmental stages, being detected in pollen mother cells. It was distributed uniformly within the cytoplasm of uni- and bicellular pollen grains and infected the generative cell. In mature pollen grains, characterized by their triangular shape, the virus was located mainly at the apertures, suggesting that PNRSV distribution follows the same pattern as the cellular components required for pollen tube germination and cell wall tube synthesis. PNRSV also was localized inside pollen tubes, especially in the growth zone. In vitro experiments demonstrated that infection with PNRSV decreases the germination percentage of pollen grains by more than half and delays the growth of pollen tubes by ≈24 h. However, although PNRSV infection affected apricot pollen grain performance during germination, the presence of the virus did not completely prevent fertilization, because the infected apricot pollen tubes, once germinated, were able to reach the apricot embryo sacs, which, in the climatic conditions of southeastern Spain, mature later than in other climates. Thus, infected pollen still could play an important role in the vertical transmission of PNRSV in apricot.

scholarly journals Pollen tube incompatibility reaction on the stigma in selfpollinated Sinapis alba L.

2014 ◽  
Vol 65 (1-2) ◽  
pp. 101-105 ◽  
Author(s):  
Renata Śnieżko ◽  
Krystyna Winiarczyk
Keyword(s):  
Pollen Tube ◽  
Pollen Tube Growth ◽  
Pollen Grains ◽  
Pollen Tubes ◽  
Sinapis Alba ◽  
Pollen Grain ◽  
Tube Growth ◽  
Sinapis Alba L ◽  
Growth Changes ◽  
Incompatibility Reaction

After selfpollination of <em>Sinapis alba</em> L. pollen tubes growth is inhibited on the stigma. The pollen grains germinate 3-4 hours after pollination. The pollen give rise to one or more pollen tubes. They grow along the papillae. In the place of contact between the papilla and pollen tube the pellicula is digested. Then the direction of pollen tube growth changes completely. Pollen tubes grow back on the exine of their own pollen grain, or turn into the air. The pollen tubes growth was inhibited in 6-8 hours after selfpollination. After crosspollination usually there is no incompatibility reaction.

scholarly journals Two groups of Arabidopsis receptor kinases preferentially regulate the growth of intraspecies pollen tubes in the female reproductive tract

2020 ◽  
Author(s):  
Hyun Kyung Lee ◽  
Daphne R. Goring
Keyword(s):  
Pollen Tube ◽  
Reproductive Tract ◽  
Cell Communication ◽  
Pollen Grains ◽  
Pollen Tubes ◽  
Female Reproductive Tract ◽  
Wild Type ◽  
Knockout Mutant ◽  
Receptor Kinases ◽  
Compatible Pollen

SummaryIn flowering plants, continuous cell-cell communication between the compatible male pollen grain/growing pollen tube and the female pistil is required for successful sexual reproduction. In Arabidopsis thaliana, the later stages of this dialogue are mediated by several peptide ligands and receptor kinases that guide pollen tubes to the ovules for the release of sperm cells. Despite a detailed understanding of these processes, a key gap remains on the nature of the regulators that function at the earlier stages. Here, we report on two groups of A. thaliana receptor kinases, the LRR-VIII-2 RK subclass and the SERKs, that function in the female reproductive tract to regulate the compatible pollen grains and early pollen tube growth, both essential steps for the downstream processes leading to fertilization. Multiple A. thaliana LRR-VIII-2 RK and SERK knockout mutant combinations were created, and several phenotypes were observed such as reduced wild-type pollen hydration and reduced pollen tube travel distances. As these mutant pistils displayed a wild-type morphology, the observed altered responses of the wild-type pollen are proposed to result from the loss of these receptor kinases leading to an impaired pollen-pistil dialogue at these early stages. Furthermore, using pollen from related Brassicaceae species, we also discovered that these receptor kinases are required in the female reproductive tract to establish a reproductive barrier to interspecies pollen. Thus, we propose that the LRR-VIII-2 RKs and the SERKs play a dual role in the preferential selection and promotion of intraspecies pollen over interspecies pollen.

scholarly journals Tradescantia bracteata pollen in vitro: pollen tubes development and mitosis

2015 ◽  
Vol 46 (4) ◽  
pp. 587-598 ◽  
Author(s):  
E. Lewandowska ◽  
M. Charzyńska
Keyword(s):  
Pollen Tube ◽  
Generative Cell ◽  
Metaphase Plate ◽  
Pollen Grains ◽  
Neutral Red ◽  
Generative Nucleus ◽  
Mitotic Spindles ◽  
Germinating Pollen ◽  
Vacuolar System

About 90 per cent of <i>Tradescantia bracteata</i> pollen germinates <i>in vitro</i> after 15 min. Mitosis starts in the pollen tube after about 3 h. The mitotic trans-formations of chromosomes within the generative nucleus are not synchronized. They involve succesively the linearly arranged chromosomes in the elongated generative nucleus. In metaphase the chromosomes are arranged tandem-like linearly along the pollen tube. The chromatides translocate in anaphase from various distances to the poles in a plane parallel to the metaphase plate. This suggests that chromosomes have individual mitotic spindles and that coordination of the chromosome transformations in the generative cell is much less strict than in a typical somatic mitosis. Starch is the storage material of pollen grains. In the vegetative cytoplasm of mature pollen grains minute reddish-orange vesicular structures are visible after staining with neutral red. They do not fuse with the vacuoles proper arising in germinating pollen grains to form the vacuolar system of the pollen tube.

scholarly journals Impact of Manganese on Pollen Germination and Tube Growth in Lily

2021 ◽  
Vol 74 ◽  
Author(s):  
Thomas Sawidis ◽  
Gülriz Baycu ◽  
Elżbieta Weryszko-Chmielewska ◽  
Aneta Sulborska
Keyword(s):  
Pollen Tube ◽  
Pollen Germination ◽  
Surrounding Medium ◽  
Pollen Grains ◽  
Lilium Longiflorum ◽  
Tube Growth ◽  
Low Concentrations ◽  
Main Effect

Abstract In vitro culture of Lilium longiflorum pollen grains was carried out to determine the role of manganese in pollen germination and pollen tube growth. Pollen germination was adversely affected by the presence of manganese (>10 −8 M), whereas low concentrations (10 −12 –10 −10 M) stimulated the process. Manganese caused morphological anomalies during tube growth, characterized by irregular pollen tube thickening and swollen tips. The main effect was the anomalous cell wall formation at the tip, in which the presence of several organelles reduced the number of secretory vesicles. A loose network of fibrillar material and spherical aggregates, mostly in the tip region, was detected, and this material was progressively loosened into the surrounding medium. As a response to potential toxicity, the excess manganese was isolated in vacuoles, which formed an internal barrier against penetration of manganese to the tip area. Elevated manganese concentrations might affect plant reproduction, resulting in anomalies in gamete development. Consequently, the loss in genetic diversity and decreased fruit set ultimately lower yield.

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