Cell division plane (CDP) orientation and cell growth directionality, determine tissue patterning and organ formation. A core component of CDP orientation is the preprophase microtubule band (PPB). It is known that PPB is physically associated with the plasma membrane (PM) requiring recruitment of scaffold proteins with affinity towards specific PM lipids and for proteins associated with microtubules. The mechanisms underlying the determination of CDP orientation are being vigorously studied. Yet, the specific identity and the role of PM lipids in the recruitment of scaffold proteins remain elusive. Sterols, a major lipid group forming nanodomains at the PM, play vital roles in cellular processes such as endocytosis, cell polarity and, cell expansion. Moreover, early studies on Arabidopsis sterol biosynthesis mutants have described severe developmental defects. However, no mechanism of implication of sterol biosynthesis deficiency in the determination of cell division has yet been proposed. Here, we have revisited the role of structural sterol biosynthesis on Arabidopsis vegetative growth and tissue patterning, as contrasted defects arising from compromised production of brassinosteroids, focusing on how CDP specific microtubule arrays are formed. Sterol biosynthesis mutants cpi1-1 and smt2smt3 showed defective vegetative phenotypes and disrupted histological pattern, as exemplified in the root. Aberrant mitosis with ectopic cell divisions. The smt2smt3 showed more severe phenotype than cpi1-1 indicating a possible role of structural phytosterols in the determination of CDP. Pharmacological treatments with sterol biosynthesis inhibitors phenocopied the sterol biosynthesis mutants. To conclude, cell division abnormalities are due to sterol biosynthesis pathway rather than brassinosteroids biosynthesis.