Identification and validation of TCONS_00093333 for regulating fertility conversion of thermo-sensitive cytoplasmic male-sterility wheat with Aegilops kotschyi cytoplasm

An increasing number of studies have shown that long non-coding RNAs (lncRNAs) play an important role in regulating plant fertility, however, they are less studied in wheat. Here, we analyzed lncRNA sequencing data and showed that the fixation carbon sequestration pathway was closely associated with pollen development and fertility conversion in KTM3315A, and eight differentially expressed genes under different fertility conditions were significantly regulated by TCONS_00093333 (designed as TaHTMAR) and transcription factors TaBBX25 and TaOBF1. Among them, TaBBX25 and TaOBF1 were located in the nucleus and expressed in the early stage of fertile anther development. Gene silencing experiments of TaHTMAR showed that TaHTMAR positively regu-lated the expression of TaBBX25 and TaOBF1 under fertile conditions, thereby reducing male fertility of KTM3315A. This study confirms the effective roles of TaHTMAR, TaBBX25, and TaOBF1 in the regulation of male fertility in wheat and provides a valuable molecular basis for hybrid wheat breeding.