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A new species of hornshark is described from northwestern Australia based on six whole specimens and a single egg case. <i>Heterodontus marshallae</i> n. sp. was previously considered to be conspecific with <i>H. zebra</i> from the Western Pacific. The new species differs from <i>H. zebra</i> in the sequence of its <i>NADH2</i> gene, several morphological characters, egg case morphology and key coloration features. Despite the coloration being similar between <i>H. marshallae</i> n. sp. and <i>H. zebra</i>, i.e., pale background with 22 dark brown bands and saddles, they differ consistently in two key aspects. Firstly, the snout of <i>H. marshallae</i> n. sp. has a dark semicircular bar, usually bifurcated for most of its length vs. a pointed, triangular shaped dark marking in <i>H. zebra</i>. Secondly, <i>H. zebra</i> has a dark bar originating below the posterior gill slits and extending onto anterior pectoral fin, which is absent in <i>H. marshallae</i> n. sp. The <i>Heterodontus marshallae</i> n. sp. is endemic to northwestern Australia and occurs in deeper waters (125–229 m) than <i>H. zebra</i> (0–143 m).