Plant LEAFY COTYLEDON (LEC) proteins, which belong to the NF-Y transcript factor B subunit, play central role in plant embryogenesis. To investigate the mechanism of microspore induced embryo in vitro in barley, a novel LEC gene, HvLEC1, was cloned from barley using the RACE method. The isolated full-length cDNA sequence of HvLEC1 was 1004 bp, containing a 597-bp open reading frame (ORF) coding for 198 amino acids. The deduced amino acid sequence of HvLEC1 contained a LEC domain located between amino acids 1 and 59. HvLEC1 can be detected in roots, stems, leaves, and the cultured microspores, and presented the highest expression in 7 days of cultured microspores. The expression of HvLEC1 at 7 days of cultured microspores was higher in barley line BI04 than that in barley line Ji19, and callus yield at 21 days of cultured microspores was also higher in BI04 than that in Ji19, which revealed that the expression level of HvLEC1 might be in a consistent with the callus yield. The expression of HvLEC1 can also be quickly up-regulated in barley root by salt stress, which suggested that the gene of HvLEC1 may be involved in not only microspore embryogenesis but also the responses to salinity stress.