Arabidopsis 3β-Hydroxysteroid Dehydrogenases/C4-Decarboxylases Are Essential for the Pollen and Embryonic Development

Jiawen Pan, Weifeng Li, Binzhao Chen, Linchuan Liu, Jianjun Zhang*, Jianming Li*

*Corresponding author for this work

Research output: Contribution to journalJournal articlepeer-review


The biosynthesis of C27–29 sterols from their C30 precursor squalene involves C24-alkylation and the removal of three methyl groups, including two at the C4 position. The two C4 demethylation reactions require a bifunctional enzyme known as 3β-hydroxysteroid dehydrogenase/C4-decarboxylase (3βHSD/D), which removes an oxidized methyl (carboxylic) group at C4 while simultaneously catalyzing the 3β-hydroxyl→3-keto oxidation. Its loss-of-function mutations cause ergosterol-dependent growth in yeast and congenital hemidysplasia with ichthyosiform erythroderma and limb defect (CHILD) syndrome in humans. Although plant 3βHSD/D enzymes were well studied enzymatically, their developmental functions remain unknown. Here we employed a CRISPR/Cas9-based genome-editing approach to generate knockout mutants for two Arabidopsis 3βHSD/D genes, HSD1 and HSD2, and discovered the male gametophytic lethality for the hsd1 hsd2 double mutation. Pollen-specific expression of HSD2 in the heterozygous hsd1 hsd2/+ mutant not only rescued the pollen lethality but also revealed the critical roles of the two HSD genes in embryogenesis. Our study thus demonstrated the essential functions of the two Arabidopsis 3βHSD/D genes in male gametogenesis and embryogenesis.

Original languageEnglish
Article number15565
Number of pages14
JournalInternational Journal of Molecular Sciences
Issue number21
Publication statusPublished - Nov 2023

Scopus Subject Areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

User-Defined Keywords

  • sterol biosynthesis
  • 3β-hydroxysteroid dehydrogenase/C4-decarboxylase
  • C4-demethylation
  • pollen development
  • embryogenesis


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