Identification of genetic differences between two species of morphologically identical Metapenaeus Genus from different environmental locations (Khaur Abdullah & Shatt Al- Arab) Southern of Iraq using (RFLP & RAPD) PCR molecular markers

Metapenaeus affinis a species from Metapenaeus genera of Penaeidae shrimps in khaur Abdullah (salty water southern of Iraq), morphologically closely related organism with suspected member in Shatt al-Arab(freshwater)also southern of Iraq. Genetic Characterization for them was study to determine the genetically differences by using (RFLP & RAPD)/PCR molecular markers.The genetic heterogeneity at the DNA level was high in the two organisms in spite of Metapenaes genus share considerable external features. In RFLP marker, voucher DAAPV F7 in 16S rRNA gene was used which was amplified then digested by using the enzymes (SmI ,TagI and HindIII). Restriction patterns of TaqI, SmI & Hind III digested voucher DAAPV F7 in 16S rRNA gene, were clearly differentiate both Metapenaeus affinis and suspected one. Size pieces which were cut result in differing size and differing numbers of patterns Whereas a species which suspected to be Metapenaeus affinis morphologically, gave different results. Overlapping patterns between different species not observed. RFLP results were confirmed the RAPD-PCR results where PCR was carried out using standers conditions for 20 random oligonucleotide primers on gDNA samples of two organisms .Out of them, twelve primers produced high reproducibility and consistent with RAPD profile and yielded monomorphic as well as polymorphic fragments ,while 8 primers yielded either weak amplifications with ambiguity or no amplification at both. The present RAPD study had shown a higher level of polymorphism in the two members of the Metapenaeus genus. 12 random primers yielded a total of 82 fragments, of which, 50 bands were polymorphic whereas 32 fragments were monomorphic. Both molecular markers (RAPD and RFLP/PCR ) were facilitated the task of distinguishing species that are morphologically close species and very useful in determining the distribution of the exotic species.