The study for understanding the molecular mechanism of miR156/529-SPL module conserved among land plants

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K15814
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 44030:Plant molecular biology and physiology-related
• Research Institution: Kochi University (2022); The University of Tokyo (2020-2021)
• Principal Investigator: Tsuzuki Masayuki 高知大学, 教育研究部総合科学系生命環境医学部門, 講師 (40845616)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: マイクロRNA / 転写因子 / ゼニゴケ / シロイヌナズナ / 遺伝子発現制御

Outline of Final Research Achievements

In this study, we focused the miR156/529-SPL transcription factor regulatory module, which controls the transition of growth phase in land plants, and aimed to understand the molecular mechanism. We performed in vitro protein synthesis and deep sequencing to identify the DNAs binding to the MpSPL2 in Marchantia polymorpha. Although the DNA sequences were identified, not clear cis regulatory motif was not obtained. On the other hand, Arabidopsis AtSPL3 and AtSPL9 were also tested and the DNA binding to those SPL homologs were successfully identified. The cis-regulatory motif was also detected by informatics analysis. We also tried to detect DNA by in vivo ChIP-seq of MpSPL2 with transformants overexpressing MpSPL2 but could not succeed in the detection. For identification of evolutionarily conserved mechanism, further studies are needed.

Free Research Field

植物遺伝子発現制御

Academic Significance and Societal Importance of the Research Achievements

本研究は、植物にとって共通する発生メカニズムである成長期の移行の制御に関して解明しようとするものであり、将来的な作物エンジニアリングなどの場面において、重要な分子メカニズムの一端の解明を目指したものである。結果としてはin vitroでのゼニゴケSPL2タンパク質に結合したDNAの検出を試み、タンパク質の合成および結合DNAの次世代シーケンシングによる検出を行うことができたが、下流遺伝子群の同定までは至らず、引き続き実験の条件検討が必要である。一方シロイヌナズナのホモログタンパク質を用いて下流遺伝子群を同定できたため、将来的な成長期移行メカニズム解明のための解析の基盤の整備は進んだと考える。