The components， which are derived from the lipid metabolism of barley malt， destabilizes the beer flavor and its shelf life in the brewing industry. The lipoxygenase 1（ LOX-1） has been approved as the key enzyme involved in lipid metabolism in barley seeds. The identification and development of null LOX-1 barley germplasm might be of interest to promote high-quality malting barley breeding. By taking use of the previously-identified null LOX-1 Chinese barley rare allelic variation， here a Kompetitive Allele Specific PCR（ KASP） based diagnostic marker was generated and subjected for a test in 633 barley landraces which originated from Shandong and Henan province. A total of eight null LOX-1 barley accessions were identified， the geographic distribution and the allele frequency were further investigated. Taken together， this work provided LOX-1 free germplasm accessions valuable for the malting barley breeding especially in conjugation with the newly-generated KASP diagnostic