Simple SummaryIndigenous cattle have extraordinary adaptation capability to diverse environments under low input production system. However, the population size is declining rapidly in Bangladesh due to massive imports of high yielding dairy breeds. The genetic diversity measures are important for assessing population architecture as well as for development of conservation strategies. The aim of this study was to investigate genetic variability and population structure of indigenous cattle genetic resources of Bangladesh using Illumina Bovine SNP50K BeadChip genotyped data. Similar to other zebu populations, low genetic diversity measures were found in Bangladeshi cattle populations. Our findings revealed their distinct genetic structure but showed low levels of genetic differentiation among the six indigenous cattle populations. Moreover, admixture and phylogenetic analysis highlighted historical gene flow among the studied populations. Altogether, our findings provide a comprehensive genomic information on indigenous cattle populations of Bangladesh that could be utilized in their future conservation and breeding research.Understanding the genetic basis of locally adapted indigenous cattle populations is essential to design appropriate strategies and programs for their genetic improvement and conservation. Here, we report genetic diversity measures, population differentiation, and structure of 218 animals sampled from six indicine cattle populations of Bangladesh. Animals were genotyped with Illumina Bovine SNP50K BeadChip along with genotyped data of 505 individuals included from 19 zebu and taurine breeds worldwide. The principal component analysis (PCA) showed clear geographic separation between taurine and indicine lineages where Bangladeshi indigenous cattle clustered with South Asian zebu populations. However, overlapped clusters in PCA, heterozygosity estimates, and Neighbor-Joining phylogenetic tree analysis revealed weak genetic differentiation among the indigenous cattle populations of Bangladesh. The admixture analysis at K = 5 and 9 suggests distinct genetic structure of the studied populations along with 1 to 4% of taurine ancestry. The effective population size suggested a limited pool of ancestors particularly for Sahiwal and North Bengal Grey cattle. In conclusion, these findings shed insights into the genetic architecture of six indigenous cattle populations of Bangladesh for the first time and suggested as distinct gene pools without potential admixture with zebu or taurine populations.