Cellulose nanowhiskers as one kind of renewable and biocompatible nanomaterials evoke much interest because of its versatility in various applications. Herein, the sisal cellulose nanowhiskers with length of 100–500 nm, ultrathin diameter of 6–61 nm, high crystallinity of 74.74 % and C6 carboxylate groups converted from C6 primary hydroxyls were prepared via a 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)/NaBr/NaClO system selective oxidization combined with mechanical homogenization. The effects of sodium hydroxide concentration in alkali pretreatment on the final sisal cellulose nanowhiskers were explored. It was found that with the increase of sodium hydroxide concentration, the sisal fiber crystalline type would change from cellulose I to cellulose II. The versatile sisal cellulose nanowhiskers would be particularly useful for applications in the nanocomposites as reinforcing phase, as well as in tissue engineering, filtration, pharmaceutical and optical industries as additives.