Abstract: The present study deal with the germination analysis of spores of different div mutants (divI, divII, divIV and divV) of Bacillus subtilis in ALA and AGFK systems and the relationship of these mutations with ger genes. Eight B. subtilis strains PY79 (wild type) and 07 different div mutants were the subject of this study. All div mutant strains produce germination defective spores. Their germination response was analyzed in both ALA and AGFK systems as well as in the presence of glucose, fructose and combination of both in ALA system. Inhibition effects of HgCl2 and NaN3 and reversal by mercaptoethanol and aqueous washing, respectively, were studied. On the basis of these results 1A292 (div IVB), 1A314-(div V) and 1A315(div V) were aligned with gerD, gerA and gerC mutations, respectively. Peptidoglycan components (hexoamines, techoic acid, diaminopimelic acid) of germination defective spores exhibit variation from wild type, hence changes in wall composition leads to defects in cell division and then spore germination. These studies support our earlier finding (the bases of this project) that div mutants (previously div IVA which was aligned to gerB , now div IVB and div V) exhibit pleiotrophic effects.