Development and Observation of Mature Megagametophyte Cell-Specific Fluorescent Markers

Chamberlin, Mark A.; Lawit, Shai J.

doi:10.1007/978-1-4939-7286-9_5

Cited by 3 publications

(1 citation statement)

References 18 publications

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“…That is why the isolated rice egg cells and zygotes have been used for PEG-Ca ++ mediated transfection and tissue culture for permanently transformed rice plants (Koiso et al 2017;Toda et al 2019). Using this technique, we may get specific cell marker lines like those in Arabidopsis (Chamberlin and Lawi 2017) to facilitate the study of rice seed biology.…”

Section: Viabilitymentioning

confidence: 99%

Isolation and Transfection of Rice Egg Cells and Zygotes for Cellular Localization

Bartley

Libault

et al. 2022

Preprint

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Due to the difficulty in accessing to gametes and zygotes in flowering plants, which are controlled in single cells deeply embedded in multiple tissues, little is known about how the initiation of plant embryogenesis may reflect or contrast from such systems in other eukaryotes. In this study we has developed an approach of isolation and transfection of rice egg cells and zygotes for cellular localization of rice cell cycle factors (KRP5, KRP4 and FB3), which opened a pathway to monitor protein expression in rice egg cells and zygotes at different developmental stages. The advantageous feature of isolated rice cells may serve as an ideal system for studying the molecular mechanism underlying the rice zygotic division to initiate seed formation.

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Section: Viabilitymentioning

confidence: 99%

Isolation and Transfection of Rice Egg Cells and Zygotes for Cellular Localization

Bartley

Libault

et al. 2022

Preprint

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Transgenic Reproductive Cell Ablation

Lawit

Chamberlin

2017

Methods in Molecular Biology

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Numerous cell ablation technologies are available and have been used in reproductive tissues, particularly for male tissues and cells. The importance of ablation of reproductive tissues is toward a fundamental understanding reproductive tissue development and fertilization, as well as, in developing sterility lines important to breeding strategies. Here, we describe techniques for developing ablation lines for both male and female reproductive cells. Also discussed are techniques for analysis, quality control, maintenance, and the lessening of pleiotropism in such lines.

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A CDKB/KRP/FB3 cell cycle core complex functions in rice gametes and zygotes

Bartley

Libault

et al. 2022

Preprint

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The cell cycle controls division and proliferation of all eukaryotic cells and is tightly regulated at multiple checkpoints by complexes of core cell cycle proteins. Due to the difficulty in accessing female gametes and zygotes of flowering plants, little is known about the molecular mechanisms underlying initiation embryogenesis despite the crucial importance of this process for seed crops. In this study, we reveal four levels of factors involved in rice zygotic cell cycle control and characterize their functions and regulation. Protein-protein interaction studies, including within zygote cells, and in vitro biochemical analyses delineate a model of the zygotic cell cycle core complex for rice. In this model, CDKB1, a major regulator of plant mitosis, is a cyclin (CYCD5)-dependent kinase; its activity is coordinately inhibited by two cell cycle inhibitors, KRP4 and KRP5; and both KRPs are regulated via F-box protein 3 (FB3)-mediated proteolysis. Supporting their critical role in controlling the rice zygotic cell cycle, mutations in KRP4, KRP5, and FB3 result in the compromised function of sperm cells and abnormal organization of female germ units, embryo and endosperm, thus significantly reducing seed-set rates. This work helps reveal regulatory mechanisms controlling the zygotic cell cycle toward seed formation in angiosperms.

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Development and Observation of Mature Megagametophyte Cell-Specific Fluorescent Markers

Cited by 3 publications

References 18 publications

Isolation and Transfection of Rice Egg Cells and Zygotes for Cellular Localization

Isolation and Transfection of Rice Egg Cells and Zygotes for Cellular Localization

Transgenic Reproductive Cell Ablation

A CDKB/KRP/FB3 cell cycle core complex functions in rice gametes and zygotes

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