Abstract

Plant membrane sterol composition has been reported to affect growth and gravitropism via polar auxin transport and auxin signaling. However, as to whether sterols influence auxin biosynthesis has received little attention. Here, by using the sterol biosynthesis mutant cyclopropylsterol isomerase1-1 (cpi1-1) and sterol application, we reveal that cycloeucalenol, a CPI1 substrate, and sitosterol, an end-product of sterol biosynthesis, antagonistically affect auxin biosynthesis. The short root phenotype of cpi1-1 was associated with a markedly enhanced auxin response in the root tip. Both were neither suppressed by mutations in polar auxin transport (PAT) proteins nor by treatment with a PAT inhibitor and responded to an auxin signaling inhibitor. However, expression of several auxin biosynthesis genes TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS1 (TAA1) was upregulated in cpi1-1. Functionally, TAA1 mutation reduced the auxin response in cpi1-1 and partially rescued its short root phenotype. In support of this genetic evidence, application of cycloeucalenol upregulated expression of the auxin responsive reporter DR5:GUS (β-glucuronidase) and of several auxin biosynthesis genes, while sitosterol repressed their expression. Hence, our combined genetic, pharmacological, and sterol application studies reveal a hitherto unexplored sterol-dependent modulation of auxin biosynthesis during Arabidopsis root elongation.

Highlights

  • Sterols are essential membrane components in all eukaryotes [1]

  • We found that cycloeucalenol upregulated the expression of DR5:GUS and auxin biosynthetic genes, whereas sitosterol repressed the expression of these genes and partially rescued the short root phenotype of cpi1-1

  • We found that the cpi1-1 mutant responds normally to the auxin signaling inhibitor, so it is unlikely that auxin signaling is enhanced in cpi1-1

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Summary

Introduction

Sterols are essential membrane components in all eukaryotes [1]. Plants have a complex sterol composition in which sitosterol, campesterol, stigmasterol, and isofucosterol are predominant [2,3,4,5]. Campesterol is the precursor for the biosynthesis of brassinosteroids (BRs), which is the only steroid hormone identified in plants so far. Whereas the dwarf phenotype of downstream sterol biosynthesis mutants, such as dwarf (dwf7)/sterol (ste1), dwf, and dwf1/diminuto (dim) could be rescued by BRs [11,12,13,14], the defect of root or hypocotyl elongation in upstream mutants such as fackel (fk)/hydra (hyd2), hyd, sterol methyltransferase (smt1), cyp51A2, cyclopropylsterol isomerase (cpi1-1), sterol 4α-methyl oxidase (smo2-1) smo2-2/+, and smo smo1-2/+. The upstream sterol mutants often display defects in auxin response and polar auxin transport (PAT). The fk/hyd and hyd mutants showed enhanced responses to auxin, and their short root and short leaf petiole phenotypes could be partially rescued by blocking auxin signaling [23]. The cotyledon vascular patterning (cvp1) mutant showed weak auxin insensitivity, and could enhance the auxin resistance of transport inhibitor resistant

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