A cell-based fluorescent system and statistical framework to detect meiosis-like induction in plants

Front Plant Sci. 2024 Jul 8:15:1386274. doi: 10.3389/fpls.2024.1386274. eCollection 2024.

Abstract

Genetic gains made by plant breeders are limited by generational cycling rates and flowering time. Several efforts have been made to reduce the time to switch from vegetative to reproductive stages in plants, but these solutions are usually species-specific and require flowering. The concept of in vitro nurseries is that somatic plant cells can be induced to form haploid cells that have undergone recombination (creating artificial gametes), which can then be used for cell fusion to enable breeding in a Petri dish. The induction of in vitro meiosis, however, is the largest current bottleneck to in vitro nurseries. To help overcome this, we previously described a high-throughput, bi-fluorescent, single cell system in Arabidopsis thaliana, which can be used to test the meiosis-like induction capabilities of candidate factors. In this present work, we validated the system using robust datasets (>4M datapoints) from extensive simulated meiosis induction tests. Additionally, we determined false-detection rates of the fluorescent cells used in this system as well as the ideal tissue source for factor testing.

Keywords: high-throughput; in vitro biology; meiosis-like induction; plant breeding; protoplasts; single-cell analysis.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. We want to thank the Iowa State University Plant Sciences Institute, RF Baker Center for Plant Breeding, and KJ Frey Chair in Agronomy for their generous support. This article is also a product of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa, Project No. IOW04714(T.L.) which is supported by USDA/NIFA and State of Iowa funds. This work was also supported by the Foundation for Food & Agriculture Research under award number CA19-SS-0000000128 (T.L.) and Hatch Project grant: USDA Hatch Project IOW03717(S.D.). The content of this manuscript is solely the responsibility of the authors and does not necessarily represent the official views of the Foundation for Food & Agriculture Research or the U.S. Department of Agriculture.