Molecular mechanism of cell fate and fertility determination of pollen sperm cells by single-cell tracking

2022 Fiscal Year Final Research Report

• Project Area: Remodeling Plant Reproduction System by Cell Fate Manipulations.
• Project/Area Number: 20H05779
• Research Category: Grant-in-Aid for Transformative Research Areas (B)
• Allocation Type: Single-year Grants
• Review Section: Transformative Research Areas, Section (III)
• Research Institution: Nagoya University
• Principal Investigator: Mizuta Yoko 名古屋大学, 高等研究院・ITbM, 特任助教 (70645142)
• Project Period (FY): 2020-10-02 – 2023-03-31
• Keywords: 花粉 / イメージング / 一過的導入 / 植物 / 生殖

Outline of Final Research Achievements

In this study, some Nicotiana benthamiana fluorescent marker lines were developed. We established a method to observe and analyze pollen and pollen tube development using confocal microscopy with flourescent markers for a long period. We also transiently introduced genes into pollen and evaluated their effects on gene function and phenotype. Our results indicate that the position of the pollen nucleus at the first division is important for the differentiation of generative cell and vegetative cell. Gene-introduced pollen grains were isolated and gene expression analysis from a small number of gene-introduced pollen grains was performed by RNA-seq analysis. Furthermore, a method for automated isolation of transiently gene-introduced pollen was established. Pollination of the isolated pollen was used to evaluate pollen tube germination and following aspects. Some of the results of this research were reported in several articles and at conferences.

Free Research Field

植物生殖

Academic Significance and Societal Importance of the Research Achievements

花粉は被子植物に共通の器官である。よって、花粉の形成過程を知ることは、被子植物の生殖を知る上で重要である。また、作物の生産性の向上や不稔性の打破など、農業や育種にも重要である。本研究では、花粉に一過的な遺伝子導入を行うことで、生きたまま花粉の発生過程における遺伝子発現や形態変化を解析できるようになった。また、遺伝子導入された花粉を選抜し受粉することで、受精までを観察可能なことが示された。本研究の手法や知見を用いることで、今後は発生、分化に関わる遺伝子の調査や、種間の比較解析の研究などが促進されると期待される。