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The role of TETRASPANINS in plant cell division orientation

Periodic Reporting for period 1 - DIVISION BELL (The role of TETRASPANINS in plant cell division orientation)

Reporting period: 2021-09-01 to 2023-08-31

The project delved into understanding the role of TETRASPANIN (TET) proteins in plant vascular development, addressing a knowledge gap in this field. Plant vascular development is essential for growth, reproduction, and response to the environment, making it vital for agriculture, ecology, and sustainability. The overall objectives were to elucidate the function of the TET proteins by a combination of genetic, biochemical and cell biological methods, while expanding the skillset and transversal competencies of the researcher, Dr. Matous Glanc.
During the project, novel tet mutant alleles were generated using CRISPR technology. A novel tet1 knockout allele was identified, which confirmed the role of TET1 in vascular proliferation and unveiled a previously unappreciated function in ground tissue patterning. This enhanced the understanding of TET functions in plant vascular development. Immunoprecipitation coupled to mass spectrometry and biotinylation-based proximity labeling methods were employed to identify TET protein interactors. Some candidates were validated, providing initial insights into TET protein interactions. Challenges in specificity and validation require further exploration. Challenges were encountered in generating functional fluorescent reporters for TET proteins. Successful complementation of the tet1-cko1 mutant with a functional reporter (HA-TET1) provided partial insights into TET subcellular localization. However, limitations in live-cell imaging and thus localization dynamics assessment persisted due to tag constraints.
The project results have been disseminated in the form of scientific articles and international conference presentations. The researcher engaged in multiple outreach events, sharing project insights and experiences with a broader audience. Social media were actively used for communication and networking. EU funding was acknowledged in all relevant publications.
The project generated novel tet mutants, contributing to the understanding of TET functions in vascular development. Initiatives on TET protein interactions were started, laying the foundation for future exploration. Overcoming challenges in reporter generation provided partial elucidation of TET subcellular localization. Enhanced knowledge of TET functions, including insights into their molecular interactions, could unveil new targets for modulating plant vascular development, which might in turn advance plant breeding and crop improvement. Societal impacts of such findings may include applications in agriculture for improved crops and wood production. In conclusion, the project has made strides in unraveling TET proteins' functions in plant vascular development, with potential future implications for agriculture and environmental sustainability. Challenges encountered emphasize the need for continued exploration in these promising research avenues.
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