PCR-RFLP ANALYSIS OF THE 16S RRNA AND ITS REGIONS IN BACTERIAL BLIGHT (XANTHOMONAS AXONOPODIS PV. PUNICAE) ACROSS POMEGRANATE FARMS IN KURDISTAN REGIONS-IRAQ.

Bacterial blight caused by Xanthomonas axonopodis pv. punicae (Xap) is a major biotic diseases in pomegranate (Punica granatum L.). In the field survey which lasted 2 year in six different geographical locations, Xap strains from the severely infected plant material collected including DuhokCcenter, around Erbil, Zaxo, south of Duhok, Akre, and Amedi. Xanthomonas was detected from infected plant material and its identity was confirmed by morphological, Microbial and Molecular Characterization. To study its genetic variability and phylogenetic relationship two important loci were targeted namely ITS region of 16s rDNA, 16S rRNA and then a PCR-RFLP was performed for PCR product of 16s rRNA loci using Hinf I, Hae III, and Rsa I restriction enzymes. The PCR-RFLP showed the genetic similarity coefficient ranging from 1.00 to 3.73, and the dendrogram grouped all tested strains in 4 clusters. The result revealed that this disease is in blooming stage in the country which was thought that has not been recorded before in pomegranate. To the best of our knowledge this is the first record of Xap. In Kurdistan/ Iraq therefore, further studies are needed to be performed to manage this hazardous bacterium

DNA Barcoding of Pomegranate (Punica granatum L.) Cultivars in Duhok Province- Kurdistan Region/ Iraq Using 18S–28S rRNA and ITS Region

Pomegranate is a tree species with a significant plant diversity, therefore molecular methods are necessary to define and verify approaches to recognize rapidly and correctly. This research looks at a genetic strategy for identifying pomegranate varieties in Duhok KRG. The approach is based on application of ITS region, PCR-RFLP, sequencing and SNP identification. For this study, 14 pomegranate accessions were taken from various regions, namely the Center of Duhok, Amedi, Akre, Zaxo, the South of Duhok, and Sulav. The PCR product of the 18S–28S rDNA intergenic spacer was 854bp, and the sequence analysis revealed a 99.94 percent similarity with other accession numbers in NCBI, demonstrating the use of the 18S–28S rDNA intergenic spacer for identifying and barcoding pomegranate cultivars. The PCR product of the ITS region was 700bp. This result was then employed for PCR-RFLP using two restriction enzymes namely RsaI GT/AC and HaeIII GG/CC which helped grouping as well as genetic similarities. This study Further involved sequencing examined genes were compared using the NCBI BLASTn tool and clustalo (Version 1.2.4) to determine gene location and SNP. According to this study, the result of PCR-RFLP revealed poor association between pomegranate physical morphology and genetic features, while SNP identification was identified between studied cultivars. Moreover, this result showed possible DNA barcoding of pomegranate cultivars under the study