The Arabidopsis thaliana Class II Formin FH13 Modulates Pollen Tube Growth

Formins are a large, evolutionarily conserved family of actin-nucleating proteins with additional roles in regulating microfilament, microtubule, and membrane dynamics. Angiosperm formins, expressed in both sporophytic and gametophytic tissues, can be divided into two subfamilies, Class I and Class II, each often exhibiting characteristic domain organization. Gametophytically expressed Class I formins have been documented to mediate plasma membrane-based actin assembly in pollen grains and pollen tubes, contributing to proper pollen germination and pollen tube tip growth, and a rice Class II formin, FH5/RMD, has been proposed to act as a positive regulator of pollen tube growth based on mutant phenotype and overexpression data. Here we report functional characterization of the Arabidopsis thaliana pollen-expressed typical Class II formin FH13 (At5g58160). Consistent with published transcriptome data, live-cell imaging in transgenic plants expressing fluorescent protein-tagged FH13 under the control of the FH13 promoter revealed expression in pollen and pollen tubes with non-homogeneous signal distribution in pollen tube cytoplasm, suggesting that this formin functions in the male gametophyte. Surprisingly, fh13 loss of function mutations do not affect plant fertility but result in stimulation of in vitro pollen tube growth, while tagged FH13 overexpression inhibits pollen tube elongation. Pollen tubes of mutants expressing a fluorescent actin marker exhibited possible minor alterations of actin organization. Our results thus indicate that FH13 controls or limits pollen tube growth, or, more generally, that typical Class II formins should be understood as modulators of pollen tube elongation rather than merely components of the molecular apparatus executing tip growth.

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

Plants ◽

10.3390/plants8060156 ◽

2019 ◽

Vol 8 (6) ◽

pp. 156 ◽

Cited By ~ 4

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.

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Arabidopsis ABCG28 is required for the apical accumulation of reactive oxygen species in growing pollen tubes

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1902010116 ◽

2019 ◽

Vol 116 (25) ◽

pp. 12540-12549 ◽

Cited By ~ 6

Author(s):

Thanh Ha Thi Do ◽

Hyunju Choi ◽

Michael Palmgren ◽

Enrico Martinoia ◽

Jae-Ung Hwang ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Pollen Tube ◽

Pollen Tube Growth ◽

Tip Growth ◽

Ectopic Expression ◽

Reactive Oxygen ◽

Pollen Tubes ◽

Tube Growth ◽

Oxygen Species ◽

Ros Accumulation

Tip-focused accumulation of reactive oxygen species (ROS) is tightly associated with pollen tube growth and is thus critical for fertilization. However, it is unclear how tip-growing cells establish such specific ROS localization. Polyamines have been proposed to function in tip growth as precursors of the ROS, hydrogen peroxide. The ABC transporter AtABCG28 may regulate ROS status, as it contains multiple cysteine residues, a characteristic of proteins involved in ROS homeostasis. In this study, we found that AtABCG28 was specifically expressed in the mature pollen grains and pollen tubes. AtABCG28 was localized to secretory vesicles inside the pollen tube that moved toward and fused with the plasma membrane of the pollen tube tip. Knocking out AtABCG28 resulted in defective pollen tube growth, failure to localize polyamine and ROS to the growing pollen tube tip, and complete male sterility, whereas ectopic expression of this gene in root hair could recover ROS accumulation at the tip and improved the growth under high-pH conditions, which normally prevent ROS accumulation and tip growth. Together, these data suggest that AtABCG28 is critical for localizing polyamine and ROS at the growing tip. In addition, this function of AtABCG28 is likely to protect the pollen tube from the cytotoxicity of polyamine and contribute to the delivery of polyamine to the growing tip for incorporation into the expanding cell wall.

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Ovary Signals for Pollen Tube Guidance in Chalazogamous Mangifera indica L.

Frontiers in Plant Science ◽

10.3389/fpls.2020.601706 ◽

2021 ◽

Vol 11 ◽

Author(s):

Jorge Lora ◽

Veronica Perez ◽

Maria Herrero ◽

Jose I. Hormaza

Keyword(s):

Pollen Tube ◽

Pollen Tube Growth ◽

Mangifera Indica ◽

Embryo Sac ◽

Pollen Tubes ◽

Tube Growth ◽

Transmitting Tissue ◽

Egg Apparatus ◽

Pollen Tube Elongation

Most flowering plants show porogamy in which the pollen tubes reach the egg apparatus through the micropyle. However, several species show chalazogamy, an unusual pollen tube growth, in which the pollen tubes reach the embryo sac through the chalaza. While ovary signals for pollen tube growth and guidance have been extensively studied in porogamous species, few studies have addressed the process in chalazogamous species such as mango (Mangifera indica L.), one of the five most important fruit crops worldwide in terms of production. In this study, we characterize pollen–pistil interaction in mango, paying special attention to three key players known to be involved in the directional pollen tube growth of porogamous species such as starch, arabinogalactan proteins (AGPs), and γ-aminobutyric acid (GABA). Starch grains were observed in the style and in the ponticulus at anthesis, but their number decreased 1 day after anthesis. AGPs, revealed by JIM8 and JIM13 antibodies, were homogenously observed in the style and ovary, but were more conspicuous in the nucellus around the egg apparatus. GABA, revealed by anti-GABA antibodies, was specifically observed in the transmitting tissue, including the ponticulus. Moreover, GABA was shown to stimulate in vitro mango pollen tube elongation. The results support the heterotrophic growth of mango pollen tubes in the style at the expense of starch, similarly to the observations in porogamous species. However, unlike porogamous species, the micropyle of mango does not show high levels of GABA and starch, although they were observed in the ponticulus and could play a role in supporting the unusual pollen tube growth in chalazogamous species.

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Cysteine-rich peptides promote interspecific genetic isolation in Arabidopsis

Science ◽

10.1126/science.aau9564 ◽

2019 ◽

Vol 364 (6443) ◽

pp. eaau9564 ◽

Cited By ~ 24

Author(s):

Sheng Zhong ◽

Meiling Liu ◽

Zhijuan Wang ◽

Qingpei Huang ◽

Saiying Hou ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Pollen Tube ◽

Reproductive Isolation ◽

Gene Family ◽

Pollen Tube Growth ◽

Pollen Tubes ◽

Flowering Plants ◽

Tube Growth ◽

Genetic Isolation ◽

Tube Emergence

Reproductive isolation is a prerequisite for speciation. Failure of communication between female tissues of the pistil and paternal pollen tubes imposes hybridization barriers in flowering plants. Arabidopsis thaliana LURE1 (AtLURE1) peptides and their male receptor PRK6 aid attraction of the growing pollen tube to the ovule. Here, we report that the knockout of the entire AtLURE1 gene family did not affect fertility, indicating that AtLURE1-PRK6–mediated signaling is not required for successful fertilization within one Arabidopsis species. AtLURE1s instead function as pollen tube emergence accelerators that favor conspecific pollen over pollen from other species and thus promote reproductive isolation. We also identified maternal peptides XIUQIU1 to -4, which attract pollen tubes regardless of species. Cooperation between ovule attraction and pollen tube growth acceleration favors conspecific fertilization and promotes reproductive isolation.

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

Horticulture Research ◽

10.1038/s41438-021-00578-z ◽

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.

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Two Expansin Genes, AtEXPA4 and AtEXPB5, Are Redundantly Required for Pollen Tube Growth and AtEXPA4 Is Involved in Primary Root Elongation in Arabidopsis thaliana

Genes ◽

10.3390/genes12020249 ◽

2021 ◽

Vol 12 (2) ◽

pp. 249

Author(s):

Weimiao Liu ◽

Liai Xu ◽

Hui Lin ◽

Jiashu Cao

Keyword(s):

Arabidopsis Thaliana ◽

Pollen Tube ◽

Pollen Tube Growth ◽

Cell Walls ◽

Root Elongation ◽

Expression Patterns ◽

Primary Root ◽

Pollen Grains ◽

Tube Growth ◽

Cell Wall Binding

The growth of plant cells is inseparable from relaxation and expansion of cell walls. Expansins are a class of cell wall binding proteins, which play important roles in the relaxation of cell walls. Although there are many members in expansin gene family, the functions of most expansin genes in plant growth and development are still poorly understood. In this study, the functions of two expansin genes, AtEXPA4 and AtEXPB5 were characterized in Arabidopsis thaliana. AtEXPA4 and AtEXPB5 displayed consistent expression patterns in mature pollen grains and pollen tubes, but AtEXPA4 also showed a high expression level in primary roots. Two single mutants, atexpa4 and atexpb5, showed normal reproductive development, whereas atexpa4atexpb5 double mutant was defective in pollen tube growth. Moreover, AtEXPA4 overexpression enhanced primary root elongation, on the contrary, knocking out AtEXPA4 made the growth of primary root slower. Our results indicated that AtEXPA4 and AtEXPB5 were redundantly involved in pollen tube growth and AtEXPA4 was required for primary root elongation.

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Testing of self-(in)compatibility in apricot cultivars from European breeding programmes

Horticultural Science ◽

10.17221/219/2012-hortsci ◽

2013 ◽

Vol 40 (No. 2) ◽

pp. 65-71 ◽

Cited By ~ 3

Author(s):

D. Milatović ◽

D. Nikolić ◽

B. Krška

Keyword(s):

Fluorescence Microscopy ◽

Pollen Tube ◽

Pollen Tube Growth ◽

Reliable Method ◽

Pollen Tubes ◽

Tube Growth ◽

Self Incompatibility ◽

Breeding Programmes

Self-(in)compatibility was tested in 40 new apricot cultivars from European breeding programmes. Pollen-tube growth in pistils from laboratory pollinations was analysed using the fluorescence microscopy. Cultivars were considered self-compatible if at least one pollen tube reached the ovary in the majority of pistils. Cultivars were considered self- incompatible if the growth of pollen tubes in the style stopped along with formation of characteristic swellings. Of the examined cultivars, 18 were self-compatible and 22 were self-incompatible. Fluorescence microscopy provides a relatively rapid and reliable method to determine self-incompatibility in apricot cultivars.      

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