Molecular characterization of Arabidopsis thaliana LSH1 and LSH2 genes
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Genes & Genomics https://doi.org/10.1007/s13258-020-00985-x
RESEARCH ARTICLE
Molecular characterization of Arabidopsis thaliana LSH1 and LSH2 genes Myungjin Lee1 · Xiangshu Dong2 · Hayong Song1 · Ju Yeon Yang1 · Soyun Kim1 · Yoonkang Hur1 Received: 20 May 2020 / Accepted: 4 August 2020 © The Genetics Society of Korea 2020
Abstract Background Arabidopsis thaliana genome encodes ten DUF640 (short for domain of unknown function 640)/ALOG (short for Arabidopsis LSH1 and Oryza G1) proteins, also known as light-dependent short hypocotyl (LSH) proteins. While some of the LSH genes regulate organ boundary determination and shade avoidance response, the function of most of these genes remains largely unknown. Objective In this study, we aimed to characterize the function of AtLSH1 and AtLSH2 in Arabidopsis. Methods We overexpressed AtLSH1 and AtLSH2 (with or without the FLAG tag) in Arabidopsis Col-0 plants under the control of the 35S promoter. We also generated knockout or knockdown lines of these genes by miRNA-induced gene silencing (MIGS). We conducted intensive phenotypic analysis of these transgenic lines, and finally performed RNA-seq analysis of two AtLSH2 overexpression (OX) lines. Results Although AtLSH1 and AtLSH2 amino acid sequences showed high similarly, AtLSH2-OX lines showed much higher levels of their transcripts than those of AtLSH1-OX lines. Additionally, overexpression of AtLSH1 and AtLSH2 greatly inhibited hypocotyl elongation in a light-independent manner, and reduced both vegetative and reproductive growth. However, knockout or knockdown of both these AtLSH genes did not affect plant phenotype. Gene Ontology (GO) analysis of differentially expressed genes (DEGs) identified by RNA-seq revealed enrichment of the GO term ‘response to stimulus’, included phytohormone-responsive genes; however, genes responsible for the abnormal phenotypes of AtLSH2-OX lines could not be identified. Conclusion Although our data revealed no close association between light and phytohormone signaling components, overexpression of AtLSH1 and AtLSH2 greatly reduced vegetative and reproductive growth of Arabidopsis plants. This property could be used to generate new plants by regulating expression of AtLSH1 and AtLSH2. Keywords DUF640/ALOG · AtLSH1 · AtLSH2 · Hypocotyl elongation · MIGS
Introduction
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13258-020-00985-x) contains supplementary material, which is available to authorized users. * Yoonkang Hur [email protected] 1
Department of Biological Sciences, College of Biological Science and Biotechnology, Chungnam National University, Daejeon 34134, Republic of Korea
School of Agriculture, Yunnam University, Kunming 650091, China
2
The light-dependent (sensitive) short hypocotyl 1 (LSH1) gene was first identified in Arabidopsis thaliana as a regulator of hypocotyl elongation; overexpression of LSH1 resulted in a hypersensitive response (short hypocotyls) in a light-dependent manner (Zhao et al. 2004). The LSH1
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