A novel actinobacterium, designated strain NEAU-D428T, was isolated from rhizosphere soil of wheat and characterized using a polyphasic approach. Morphological and chemotaxonomic characteristics of the strain coincided with members of the genus
Microbispora
. The 16S rRNA gene sequence analysis showed that the isolate was most closely related to
Microbispora bryophytorum
NEAU-TX2-2T (99.2 %). Phylogenetic analysis based on the 16S rRNA gene sequences indicated that the strain clustered with
Microbispora clausenae
CLES2T (99.1 %), but formed a separate subclade in the phylogenomic tree within the genus
Microbispora
. The menaquinones were identified as MK-9(H4), MK-9(H2) and MK-9(H0). The phospholipid profile was found to consist of diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, ninhydrin-positive glycophospholipid, phosphatidylinositol and phosphatidylinositol mannoside. The major fatty acids were identified as iso-C16 : 0, C16 : 0, 10-methyl C17 : 0 and C18 : 0. Digital DNA–DNA hybridization and average nucleotide identity values between strain NEAU-D428T and
M. bryophytorum
NEAU-TX2-2T,
Microbispora camponoti
2C-HV3T,
M. clausenae
CLES2T, ‘Microbispora cellulosiformans’ Gxj-6T and
Microbispora fusca
NEAU-HEGS1-5T were 47.6 and 92.2 %, 47.5 and 92.2 %, 55.9 and 94.0 %, 33.1 and 86.8 %, and 33.6 and 87.1 %, respectively. These results and some physiological and biochemical properties demonstrated that the strain could be distinguished from its closest relatives. Therefore, it is proposed that strain NEAU-D428T should be classified as representative of a novel species of the genus
Microbispora
, for which the name Microbispora sitophila sp. nov. is proposed. The type strain is NEAU-D428T (=CGMCC 4.7523T=DSM 109822T).