Frontiers in Plant Science (Aug 2022)

Functional analysis of Pogostemon cablin farnesyl pyrophosphate synthase gene and its binding transcription factor PcWRKY44 in regulating biosynthesis of patchouli alcohol

  • Xiaobing Wang,
  • Xiaobing Wang,
  • Xiaobing Wang,
  • Yun Tang,
  • Yun Tang,
  • Yun Tang,
  • Huiling Huang,
  • Huiling Huang,
  • Huiling Huang,
  • Daidi Wu,
  • Daidi Wu,
  • Daidi Wu,
  • Xiuzhen Chen,
  • Xiuzhen Chen,
  • Xiuzhen Chen,
  • Xiuzhen Chen,
  • Junren Li,
  • Junren Li,
  • Junren Li,
  • Junren Li,
  • Hai Zheng,
  • Ruoting Zhan,
  • Ruoting Zhan,
  • Ruoting Zhan,
  • Ruoting Zhan,
  • Likai Chen,
  • Likai Chen,
  • Likai Chen,
  • Likai Chen

DOI
https://doi.org/10.3389/fpls.2022.946629
Journal volume & issue
Vol. 13

Abstract

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Farnesyl pyrophosphate synthase (FPPS) plays an important role in the synthesis of plant secondary metabolites, but its function and molecular regulation mechanism remain unclear in Pogostemon cablin. In this study, the full-length cDNA of the FPP synthase gene from P. cablin (PcFPPS) was cloned and characterized. The expressions of PcFPPS are different among different tissues (highly in P. cablin flowers). Subcellular localization analysis in protoplasts indicated that PcFPPS was located in the cytoplasm. PcFPPS functionally complemented the lethal FPPS deletion mutation in yeast CC25. Transient overexpression of PcFPPS in P. cablin leaves accelerated terpene biosynthesis, with an ~47% increase in patchouli alcohol. Heterologous overexpression of PcFPPS in tobacco plants was achieved, and it was found that the FPP enzyme activity was significantly up-regulated in transgenic tobacco by ELISA analysis. In addition, more terpenoid metabolites, including stigmasterol, phytol, and neophytadiene were detected compared with control by GC-MS analysis. Furthermore, with dual-LUC assay and yeast one-hybrid screening, we found 220 bp promoter of PcFPPS can be bound by the nuclear-localized transcription factor PcWRKY44. Overexpression of PcWRKY44 in P. cablin upregulated the expression levels of PcFPPS and patchoulol synthase gene (PcPTS), and then promote the biosynthesis of patchouli alcohol. Taken together, these results strongly suggest the PcFPPS and its binding transcription factor PcWRKY44 play an essential role in regulating the biosynthesis of patchouli alcohol.

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