Fragaria nilgerrensis
is a wild diploid strawberry species endemic to east and southeast region in Asia and provides a rich source of genetic variations for strawberry improvement. Here, we present a chromosome‐scale assembly of
F. nilgerrensis
using single‐molecule real‐time (SMRT) Pacific Biosciences sequencing and chromosome conformation capture (Hi‐C) genome scaffolding. The genome assembly size was 270.3 Mb, with a contig N50 of ∼8.5 Mb. A total of 28 780 genes and 117.2 Mb of transposable elements were annotated for this genome. Next, detailed comparative genomics with the high‐quality
F. vesca
reference genome was conducted to obtain the difference among transposable elements, SNPs, Indels, and so on. The genome size of
F. nilgerrensis
was enhanced by around 50 Mb relatively to
F. vesca
, which is mainly due to expansion of transposable elements. In comparison with the
F
.
vesca
genome, we identified 4 561 825 SNPs, 846 301 Indels, 4243 inversions, 35 498 translocations and 10 099 relocations. We also found a marked expansion of genes involved in phenylpropanoid biosynthesis, starch and sucrose metabolism, cyanoamino acid metabolism, plant–pathogen interaction, brassinosteroid biosynthesis and plant hormone signal transduction in
F. nilgerrensis
, which may account for its specific phenotypes and considerable environmental adaptability. Interestingly, we found sequence variations in the upstream regulatory region of
FnMYB10
, a core transcriptional activator of anthocyanin biosynthesis, resulted in the low expression level of the
FnMYB10
gene, which is likely responsible for white fruit phenotype of
F. nilgerrensis.
The high‐quality
F. nilgerrensis
genome will be a valuable resource for biological research and comparative genomics research.