Heading date and growth habit are a key factor that regulates the transition from vegetative to reproductive stage in barley. In this study we used PCR based markers to identify the allelic variations at Vrn-H1 (HvMB5) and Vrn-H2 (HvSNF2) genes and to predict the heading date and growth habit of a collection of Tunisian barley assessed under semi-arid climate. Allelic variation at HvBM5 revealed two PCR fragments 830bp and 344 bp. Primer sets used to amplify HvSNF2 gene have resulted different alleles size of 543bp, 623bp and 700bp. Different allelic combination of HVBM5 and HvSNF2 were associated with heading date and growth habit. Spring and early heading accessions were only characterized by the amplification of HvSNF2 fragment 700bp. All winter accessions have yielded the PCR product HvBM5 830bp, but the variation of heading date was determined by the HvSNF2 alleles. These DNA markers will be a powerful tool to predict the heading date and growth habit and be used as markers for assisted selection to speed the national breeding program.