*Emails: #qijuan@sdau.edu.cn, *dlfu@sdau.edu.cn, *nifei1998@sdau.edu.cn.

Plant male sterility is a valuable trait, that is widely exploited in hybrid seed production. This trait can be classified into genic male sterility (GMS) and cytoplasmic male sterility (CMS). Poor genetic diversity, unreliable restoration, and a potential cytoplasmic penalty on heterosis limited the application of CMS to hybrid seed production.

The use of GMS can overcome these drawbacks. The barley accession ‘GSHO3002’ is a spontaneous male sterile mutant in barley variety ‘Freja/Nepal’ (CIho 15839). The barley sterility in GSHO3002 was controlled by a recessive GMS gene, msg3002 (originally named as msg,,by). In this study, by genetic mapping, we mapped MSG3002 to a 0.2 cM interval between two flanking marker SP5M21 and SP5M14 on the chromosome arm 1HL.

The SP5M21-SP5M14 interval corresponds 371.2 kb in the Morex reference sequence. This interval involves four high-confidence candidates, and sequence analysis reveals that one of them, a gene encoding the GDSL lipase (named HvGDSL64460), has a premature stop codon in the GSHO3002, which identified as the most plausible candidate for MSG3002. Afterwards, the expression profiles revealed that HvGDSL64460 was specifically expressed in anther.

The anther-specific GDSL lipase was localized in endoplasmic reticulum (ER). For validation experiments, allelic variations of HvGDSL64460 were identified from barley EMS-mutagenized population and combined with genetic analysis revealed that this anther-specific GDSL lipase is responsible for male fertility in barley. Furthermore, widely untargeted metabolomics analysis revealed that MSG3002 may function through cutin synthases/acyltransferase activity to modulate lipid metabolism.

Meanwhile, we demonstrated the evolutionary conserved role of MSG3002 in male fertility control between barley and wheat by knocking out the MSG3002 orthologs in the A, B and D subgenomes of allohexaploid wheat. Taken together, our study suggests that MSG3002 is an indispensable evolutionary conserved member in anther development of barley and wheat that could be a valuable target for GMS-based system for hybrid breeding barley and wheat.