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Molecular characterization of Arabidopsis thaliana LSH1 and LSH2 genes

Genes & Genomics 2020년 42권 10호 p.1151 ~ 1162
이명진, Dong Xiangshu, 송하용, 양주연, 김소윤, 허윤강,
소속 상세정보
이명진 ( Lee Myung-Jin ) - Chungnam National University College of Biological Science and Biotechnology Department of Biological Sciences
 ( Dong Xiangshu ) - Yunnam University School of Agriculture
송하용 ( Song Ha-Yong ) - Chungnam National University College of Biological Science and Biotechnology Department of Biological Sciences
양주연 ( Yang Ju-Yeon ) - Chungnam National University College of Biological Science and Biotechnology Department of Biological Sciences
김소윤 ( Kim So-Yun ) - Chungnam National University College of Biological Science and Biotechnology Department of Biological Sciences
허윤강 ( Hur Yoon-Kang ) - Chungnam National University College of Biological Science and Biotechnology Department of Biological Sciences

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.

키워드

DUF640/ALOG; AtLSH1; AtLSH2; Hypocotyl elongation; MIGS

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