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 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 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 NETWORKED2‐Subfamily Proteins Regulate the Cortical Actin Cytoskeleton of Growing Pollen Tubes and Polarised Pollen Tube Growth

2021 ◽  
Author(s):  
Patrick Duckney ◽  
Johan T. Kroon ◽  
Martin R. Dixon ◽  
Timothy J. Hawkins ◽  
Michael J. Deeks ◽  
...  
Keyword(s):  
Pollen Tube ◽  
Actin Cytoskeleton ◽  
Pollen Tube Growth ◽  
Pollen Tubes ◽  
Tube Growth ◽  
Cortical Actin

Pronounced cytoplasmic pH gradients are not required for tip growth in plant and fungal cells

1997 ◽  
Vol 110 (10) ◽  
pp. 1187-1198 ◽  
Author(s):  
R.M. Parton ◽  
S. Fischer ◽  
R. Malho ◽  
O. Papasouliotis ◽  
T.C. Jelitto ◽  
...  
Keyword(s):  
Tip Growth ◽  
Cell Types ◽  
Pollen Tubes ◽  
Fine Tuning ◽  
Cytoplasmic Ph ◽  
Dual Emission ◽  
Ph Gradients ◽  
Longitudinal Gradients ◽  
External Ph

The existence of pronounced cytoplasmic pH gradients within the apices of tip-growing cells, and the role of cytoplasmic pH in regulating tip growth, were investigated in three different cell types: vegetative hyphae of Neurospora crassa; pollen tubes of Agapanthus umbellatus; and rhizoids of Dryopteris affinis gametophytes. Examination of cytoplasmic pH in growing cells was performed by simultaneous, dual emission confocal ratio imaging of the pH-sensitive probe carboxy SNARF-1. Considerable attention was paid to the fine tuning of dye loading and imaging parameters to minimise cellular perturbation and assess the extent of dye partitioning into organelles. With optimal conditions, cytoplasmic pH was measured routinely with a precision of between +/−0.03 and +/−0.06 of a pH unit and a spatial resolution of 2.3 microm2. Based on in vitro calibration, estimated values of mean cytoplasmic pH for cells loaded with dye-ester were between 7.15 and 7.25 for the three cell types. After pressure injecting Neurospora hyphae with dextran-conjugated dye, however, the mean cytoplasmic pH was estimated to be 7.57. Dextran dyes are believed to give a better estimate of cytoplasmic pH because of their superior localisation and retention within the cytosol. No significant cytoplasmic pH gradient (delta pH of >0.1 unit) was observed within the apical 50 microm in growing cells of any of the three cell types. Acidification or alkalinisation of the cytoplasm in Neurospora hyphae, using a cell permeant weak acid (propionic acid at pH 7.0) or weak base (trimethylamine at pH 8.0), slowed down but did not abolish growth. However, similar manipulation of the cytoplasmic pH of Agapanthus pollen tubes and Dryopteris rhizoids completely inhibited growth. Modification of external pH affected the growth pattern of all cell types. In hyphae and pollen tubes, changes in external pH were found to have a small transient effect on cytoplasmic pH but the cells rapidly readjusted towards their original pH. Our results suggest that pronounced longitudinal gradients in cytoplasmic pH are not essential for the regulation of tip growth.

Morphology of Cell Injury: An Approach to the EDIT Programme by the Use of Tobacco Pollen Tubes

2002 ◽  
Vol 30 (3) ◽  
pp. 323-329 ◽  
Author(s):  
Udo Kristen ◽  
Natalie Bischoff ◽  
Saskia Lisboa ◽  
Enno Schirmer ◽  
Sören Witt ◽  
...  
Keyword(s):  
Tip Growth ◽  
Cell Injury ◽  
Cytosolic Calcium ◽  
Pollen Tubes ◽  
In Vitro Cytotoxicity ◽  
Toxic Compounds ◽  
In Vitro System ◽  
Tobacco Pollen ◽  
Different Types

Tobacco pollen tubes were used as a standard in vitro system to investigate cell growth aberrations caused by some of the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) programme chemicals and other toxic compounds. Changes in cytoskeletal pattern were observed in the tube cells by using tubu-lin immunofluorescence and rhodamin–phalloidin fluorescence for the localisation of microtubules and actin filaments, respectively. Four different types of cell malformation were found: screw-like growth, isodiametric tip swelling, hook formation, and pollen grain enlargement. We suggest that these malformations resulted from an interference by the chemicals with the cytosolic calcium gradient which controls tip growth and the orientation of the pollen tube. The results may contribute to a general understanding of toxicity-based cell malformations.

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 Signaling in Pollen Tube Growth: Beyond the Tip of the Polarity Iceberg

Plants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 156 ◽  
Author(s):  
Nolan Scheible ◽  
Andrew McCubbin
Keyword(s):  
Pollen Tube ◽  
Signaling Pathways ◽  
Pollen Tube Growth ◽  
Tip Growth ◽  
Holistic Approach ◽  
Pollen Tubes ◽  
Small Gtpase ◽  
Tube Growth ◽  
Concerted Action ◽  
Current State

The coordinated growth of pollen tubes through floral tissues to deliver the sperm cells to the egg and facilitate fertilization is a highly regulated process critical to the Angiosperm life cycle. Studies suggest that the concerted action of a variety of signaling pathways underlies the rapid polarized tip growth exhibited by pollen tubes. Ca2+ and small GTPase-mediated pathways have emerged as major players in the regulation of pollen tube growth. Evidence suggests that these two signaling pathways not only integrate with one another but also with a variety of other important signaling events. As we continue to elucidate the mechanisms involved in pollen tube growth, there is a growing importance in taking a holistic approach to studying these pathways in order to truly understand how tip growth in pollen tubes is orchestrated and maintained. This review considers our current state of knowledge of Ca2+-mediated and GTPase signaling pathways in pollen tubes, how they may intersect with one another, and other signaling pathways involved. There will be a particular focus on recent reports that have extended our understanding in these areas.

Unilateral Cross-Incompatibility in Eucalyptus: the Case of Hybridisation Between E. globulus and E. nitens

1990 ◽  
Vol 38 (4) ◽  
pp. 383 ◽  
Author(s):  
PL Gore ◽  
BM Potts ◽  
PW Volker ◽  
J Megalos
Keyword(s):  
Pollen Tube ◽  
Pollen Tube Growth ◽  
Pollen Tubes ◽  
Full Length ◽  
Tube Length ◽  
Style Length ◽  
Similar Difference ◽  
Structural Barrier ◽  
Pollen Tube Length

The growth of E. globulus and E. nitens pollen tubes in styles of E. globulus was examined in order to elucidate the site of the unilateral barrier to hybridisation. Pollen tubes of E. nitens failed to grow the full length of the larger E. globulus style. E. globulus pollen tubes grew an average of 1.4 mm per day for the first 4 days, compared with 0.8 mm per day for pollen tubes of E. nitens. From days 4 to 14, the growth of E. nitens pollen tubes slowed to an average of 0.2 mm per day and virtually no growth occurred after day 14. In contrast, E. globulus pollen tubes grew through the style and into the ovary between days 5 and 14. By day 28, at about the time of style abscission, E. nitens tubes had grown only 6 mm, well short of the full length of the E. globulus style (9-10 mm). A similar difference in growth was obtained in vitro where E. nitens pollen tubes were significantly shorter than those of E. globulus. A comparison also including E. ovata, E. urnigera and E. gunnii indicated a significant correlation between style length and in vitro pollen tube length. It is argued that the unilateral cross-incompatibility between E. globulus and E. nitens is due to a structural barrier arising from an inherent limit to pollen tube growth which is associated with pistil size.

Functional genomics of pollen tube–pistil interactions in Arabidopsis

2010 ◽  
Vol 38 (2) ◽  
pp. 593-597 ◽  
Author(s):  
Ravishankar Palanivelu ◽  
Mark A. Johnson
Keyword(s):  
Pollen Tube ◽  
Regulatory Networks ◽  
Genomic Analysis ◽  
Pollen Tubes ◽  
Cellular Growth ◽  
Functional Genomic ◽  
Reverse Genetic ◽  
Guidance Cues ◽  
Functional Genomic Analysis

The pollen tube represents an attractive model system for functional genomic analysis of the cell–cell interactions that mediate guided cellular growth. The pollen tube extends through pistil tissues and responds to guidance cues that direct the tube towards an ovule, where it releases sperm for fertilization. Pollen is readily isolated from anthers, where it is produced, and can be induced to produce a tube in vitro. Interestingly, pollen tube growth is significantly enhanced in pistils, and pollen tubes are rendered competent to respond to guidance cues after growth in a pistil. This potentiation of the pollen tube by the pistil suggested that pollen tubes alter their gene-expression programme in response to their environment. Recently, the transcriptomes of pollen tubes grown in vitro or through pistil tissues were determined. Significant changes in the transcriptome were found to accompany growth in vitro and through the pistil tissues. Reverse genetic analysis of pollen-tube-induced genes identified a new set of factors critical for pollen tube extension and navigation of the pistil environment. Recent advances reviewed in the present paper suggest that functional genomic analysis of pollen tubes has the potential to uncover the regulatory networks that shape the genetic architecture of the pollen tube as it responds to migratory cues produced by the pistil.

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