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Karyotype analysis was carried out on gill cells of three species of octopods using a conventional air-drying method. The karyotype results showed that all the three species have the same diploid chromosome number, 2n=60, but with different karyograms as 2n=38M+6SM+8ST+8T, FN (fundamental number)=104 (   Cistopus   chinensis    Zheng et al., 2012), 2n=42M+6SM+4ST+8T, FN=108 (   Octopus   minor    (Sasaki, 1920)) and 2n=32M+16SM+12T, FN=108 (   Amphioctopus   fangsiao    (d’Orbigny, 1839–1841)). These findings were combined with data from earlier studies to infer the genetic relationships between nine species via cluster analysis using the karyotype evolutionary distance ( D e  ) and resemblance-near coefficient ( λ ). The resulting tree revealed a clear distinction between different families and orders which was substantially consistent with molecular phylogenies. The smallest intraspecific evolutionary distance ( D e  =0.2013, 0.2399) and largest resemblance-near coefficient ( λ =0.8184, 0.7871) appeared between    O.   minor    and    C.   chinensis   , and    Sepia   esculenta    Hoyle, 1885 and    S.   lycidas    Gray, 1849, respectively, indicating that these species have the closest relationship. The largest evolutionary gap appeared between species with complicated karyotypes and species with simple karyotypes. Cluster analysis of  D e   and  λ  provides information to supplement traditional taxonomy and molecular systematics, and it would serve as an important auxiliary for routine phylogenetic study.

Comparison of the genetic relationship between nine Cephalopod species based on cluster analysis of karyotype evolutionary distance