AUTHOR=Yun Hee Rang , Chen Chong , Kim Jee Hye , Kim Hae Eun , Karthik Sivabalan , Kim Hye Jeong , Chung Young-Soo , Baek Hee Soon , Sung Sibum , Kim Hyun Uk , Heo Jae Bok 

TITLE=Genome-edited HEADING DATE 3a knockout enhances leaf production in Perilla frutescens

JOURNAL=Frontiers in Plant Science

VOLUME=14

YEAR=2023

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2023.1133518

DOI=10.3389/fpls.2023.1133518

ISSN=1664-462X

ABSTRACT=<p>Environmental cues regulate the transition of many plants from vegetative to flowering development. Day length, or photoperiod, is one cue that synchronizes flowering by changing seasons. Consequently, the molecular mechanism of flowering control is prominent in Arabidopsis and rice, where essential genes like <italic>FLOWERING LOCUS</italic> T (<italic>FT</italic>) homolog, <italic>HEADING DATE</italic> 3a (<italic>Hd3a</italic>), have been connected to flowering regulation. Perilla is a nutrient-rich leaf vegetable, and the flowering mechanism remains largely elusive. We identified flowering-related genes under short-day conditions using RNA sequencing to develop an enhanced leaf production trait using the flowering mechanism in the perilla. Initially, an <italic>Hd3a</italic>-like gene was cloned from the perilla and defined as <italic>PfHd3a</italic>. Furthermore, <italic>PfHd3a</italic> is highly rhythmically expressed in mature leaves under short-day and long-day conditions. Ectopic expression of <italic>PfHd3a</italic> in <italic>Atft-1</italic> mutant plants has been shown to complement Arabidopsis <italic>FT</italic> function, resulting in early flowering. In addition, our genetic approaches revealed that overexpression of <italic>PfHd3a</italic> in perilla caused early flowering. In contrast, the CRISPR/Cas9 generated <italic>PfHd3a</italic>-mutant perilla showed significantly late flowering, resulting in approximately 50% leaf production enhancement compared to the control. Our results suggest that <italic>PfHd3a</italic> plays a vital role in regulating flowering in the perilla and is a potential target for molecular breeding in the perilla.</p>