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Rao’s quadratic diversity index is one of the most widely applied diversity indices in functional and phylogenetic ecology. The standard way of computing Rao’s quadratic diversity index for an ecological assemblage with a group of species with varying abundances is to sum the functional or phylogenetic distances between a pair of species in the assemblage, weighted by their relative abundances. Here, using both theoretically derived and observed empirical datasets, we show that this standard calculation routine in practical applications will statistically underestimate the true value, and the bias magnitude is derived accordingly. The underestimation will become worse when the studied ecological community contains more species or the pairwise species distance is large. For species abundance data measured using the number of individuals, we suggest calculating the unbiased Rao’s quadratic diversity index.

Evaluation of the estimate bias magnitude of the Rao’s quadratic diversity index