Interactions between sterol biosynthesis genes in embryonic development of <i>Arabidopsis</i>

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<jats:title>Summary</jats:title><jats:p>The sterol biosynthesis pathway of <jats:italic>Arabidopsis</jats:italic> produces a large set of structurally related phytosterols including sitosterol and campesterol, the latter being the precursor of the brassinosteroids (BRs). While BRs are implicated as phytohormones in post‐embryonic growth, the functions of other types of steroid molecules are not clear. Characterization of the <jats:italic>fackel</jats:italic> (<jats:italic>fk</jats:italic>) mutants provided the first hint that sterols play a role in plant embryogenesis. <jats:italic>FK</jats:italic> encodes a sterol C‐14 reductase that acts upstream of all known enzymatic steps corresponding to BR biosynthesis mutants. Here we report that genetic screens for <jats:italic>fk</jats:italic>‐like seedling and embryonic phenotypes have identified two additional genes coding for sterol biosynthesis enzymes: CEPHALOPOD (CPH), a C‐24 sterol methyl transferase, and HYDRA1 (HYD1), a sterol C‐8,7 isomerase. We describe genetic interactions between <jats:italic>cph</jats:italic>, <jats:italic>hyd1</jats:italic> and <jats:italic>fk</jats:italic>, and studies with 15‐azasterol, an inhibitor of sterol C‐14 reductase. Our experiments reveal that <jats:italic>FK</jats:italic> and <jats:italic>HYD1</jats:italic> act sequentially, whereas <jats:italic>CPH</jats:italic> acts independently of these genes to produce essential sterols. Similar experiments indicate that the BR biosynthesis gene <jats:italic>DWF1</jats:italic> acts independently of <jats:italic>FK</jats:italic>, whereas BR receptor gene <jats:italic>BRI1</jats:italic> acts downstream of <jats:italic>FK</jats:italic> to promote post‐embryonic growth. We found embryonic patterning defects in <jats:italic>cph</jats:italic> mutants and describe a GC–MS analysis of <jats:italic>cph</jats:italic> tissues which suggests that steroid molecules in addition to BRs play critical roles during plant embryogenesis. Taken together, our results imply that the sterol biosynthesis pathway is not a simple linear pathway but a complex network of enzymes that produce essential steroid molecules for plant growth and development.</jats:p>

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