Fundamental understanding of phenylpropanoid, lignin and flavonoid biosynthesis regulated by metabolon formation in grasses for precise manipulation to improve biomass utility

2022 Fiscal Year Research-status Report

• Project/Area Number: 22K20590
• Research Institution: Akita University
• Principal Investigator: Lam PuiYing 秋田大学, 理工学研究科, 助教 (60961711)
• Project Period (FY): 2022-08-31 – 2024-03-31
• Keywords: lignin / flavonoids / phenylpropanoids / metabolon formation / grasses

Outline of Annual Research Achievements

The proposed research is aimed to understand the global organization of metabolons (protein-protein interaction networks) of phenylpropanoid, lignin and flavonoid biosynthetic enzymes that governs metabolite channelling and crosstalk between these pathways in grasses using rice as a model, and to elucidate how metabolon formation controls metabolic channeling in phenylpropanoid, lignin and flavonoid biosynthesis. Through in silico analysis, all the putative and known enzymes in the lignin, flavonoid and phenylpropanoid biosynthetic pathways of rice have been identified. The proposed experiments are currently ongoing.

Current Status of Research Progress

4: Progress in research has been delayed.

Reason

The proposed research is delayed because of the delay of the approval of experiments at Akita University and the delayed arrival of the plant growth chamber due to the global shortage of semi-conductor. After the approval of experiments in March, 2023, we are now obtaining the plasmids, yeasts and Agrobacterium strains essential for the proposed research. Cloning is currently ongoing. From this point of view, now the project starts to proceed as planned.

Strategy for Future Research Activity

The proposed research will be carried out as described in the proposal with slight modifications based on the newest methods reported recently. After finishing cloning, subcellular localization and yeast-two hybrid analysis of the enzymes in the phenylpropanoid, lignin and flavonoid biosynthetic pathways of rice will be carried out. A newly available split GAL4 RUBY (a series of betanidin biosynthetic genes) assay instead of bimolecular fluorescence complementation (BiFC) will be used to examine protein-protein interaction in planta. Unlike BiFC that requires confocal microscope, the split GAL4 RUBY assay is more convenient as it generates visible red colour when proteins interact. Analysis of metabolite channelling will be conducted after the organization of metabolon is determined.

Causes of Carryover

In the next fiscal year, the funding will be mainly used for buying consumables and DNA sequencing service required for the proposed experiments.