Dataset Information

Analysis of gamete membrane dynamics during double fertilization of Arabidopsis.

ABSTRACT: Angiosperms have a unique sexual reproduction system called "double fertilization." One sperm cell fertilizes the egg and another sperm cell fertilizes the central cell. To date, plant gamete membrane dynamics during fertilization has been poorly understood. To analyze this unrevealed gamete subcellular behavior, live cell imaging analyses of Arabidopsis double fertilization were performed. We produced female gamete membrane marker lines in which fluorescent proteins conjugated with PIP2a finely visualized egg cell and central cell surfaces. Using those lines together with a sperm cell membrane marker line expressing GCS1-GFP, the double fertilization process was observed. As a result, after gamete fusion, putative sperm plasma membrane GFP signals were occasionally detected on the egg cell surface adjacent to the central cell. In addition, time-lapse imaging revealed that GCS1-GFP signals entered both the egg cell and the central cell in parallel with the sperm cell movement toward the female gametes during double fertilization. These findings suggested that the gamete fusion process based on membrane dynamics was composed of (1) plasma membrane fusion on male and female gamete surfaces, (2) entry of sperm internal membrane components into the female gametes, and (3) plasmogamy.

SUBMITTER: Igawa T

PROVIDER: S-EPMC4194012 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature

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Analysis of gamete membrane dynamics during double fertilization of Arabidopsis.

Igawa Tomoko T Yanagawa Yuki Y Miyagishima Shin-ya SY Mori Toshiyuki T

Journal of plant research 20121018 3

Angiosperms have a unique sexual reproduction system called "double fertilization." One sperm cell fertilizes the egg and another sperm cell fertilizes the central cell. To date, plant gamete membrane dynamics during fertilization has been poorly understood. To analyze this unrevealed gamete subcellular behavior, live cell imaging analyses of Arabidopsis double fertilization were performed. We produced female gamete membrane marker lines in which fluorescent proteins conjugated with PIP2a finely ...[more]

PMID: 23076439

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Project description:During double fertilization in angiosperms, two male gametes (sperm cells), are released from a pollen tube into the receptive region between two female gametes; the egg cell and the central cell of the ovule. The sperm cells fertilize the egg cell and the central cell in a one-to-one manner to yield a zygote and an endosperm, respectively. The one-to-one distribution of the sperm cells to the two female gametes is strictly regulated, possibly via communication among the four gametes. Polyspermy block is the mechanism by which fertilized female gametes prevent fertilization by a secondary sperm cell, and has been suggested to operate in the egg cell rather than the central cell. However, whether the central cell also has the ability to avoid polyspermy during double fertilization remains unclear. Here, we assessed the one-to-one fertilization mechanism of the central cell by laser irradiation of the female gametes and live cell imaging of the fertilization process in Arabidopsis thaliana. We successfully disrupted an egg cell within the ovules by irradiation using a femtosecond pulse laser. In the egg-disrupted ovules, the central cell predominantly showed single fertilization by one sperm cell, suggesting that neither the egg cell nor its fusion with one sperm cell is necessary for one-to-one fertilization (i.e., monospermy) of the central cell. In addition, using tetraspore mutants possessing multiple sperm cell pairs in one pollen, we demonstrated that normal double fertilization was observed even when excess sperm cells were released into the receptive region between the female gametes. In ovules accepting four sperm cells, the egg cell never fused with more than one sperm cell, whereas half of the central cells fused with more than one sperm cell (i.e., polyspermy) even 1 h later. Our results suggest that the central cell can block polyspermy during double fertilization, although the central cell is more permissive to polyspermy than the egg cell. The potential contribution of polyspermy block by the central cell is discussed in terms of how it is involved in the one-to-one distribution of the sperm cells to two distinct female gametes.

Dataset Information

Analysis of gamete membrane dynamics during double fertilization of Arabidopsis.

Publications

Analysis of gamete membrane dynamics during double fertilization of Arabidopsis.

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