Análisis de diversidad genética y secuencias de nucleótidos del marcador mildi polvoso y del gen de resistencia Vf2RAD en variedades locales de manzana (Malus domestica)

Introduction: DNA sequencing-based methods and nucleotide sequence analysis have become the most common molecular approaches currently used for molecular typing purposes and phylogenetic diversity analysis. Methods: In this study, the nucleotid sequence variations of Powdery mildew resistance gene marker (CH03c02) and the apple scab resistance gene (Vf2RAD) beside phylogenetic diversity of seven apple landraces have been investigated. The two-locus have been successfully cloned and their nucleotide sequences were determined across all studied landraces. Results: Results of sequence alignment of the Powdery mildew resistant locus (CH03c02), compared with that of the published sequence of the same locus of Discovery genotype (HiDRAS),revealed that the nucleotide variations of this locus ranged from 1 to 28 nucleotide substitutions across all seven apple landraces. Whilst, the nucleotide variations of VF2RAD ranged from 2-8 nucleotide substitutions across all the investigated landraces. The highest genetic distance (0.062) was between Amara and Barwari. Whereas, the lowest genetic distance (0.0015) was found between each of the Lubnani, Rechard, Ispartal, and the Ahmadagha. Thenucleotide sequences of the two loci were concatenated and implemented to build a Neighbor-Joining tree. The seven apple landraces were successfully grouped into two main genetic clusters (C1 and C2) in the phylogenetic tree. Conclusions: It can be concluded that the cloning approach used in the current study was found to be very successful and helpful for obtaining the full nucleotide sequences of these two loci. The investigated loci were displayed nucleotide variations among the studied landraces. And, finding of these variations was allowed the distinguishing and discrimination of these landraces

Aflp Assessment of Ficus Cultivars for Identification and Conservation

The fig (Ficus carica L., Moraceae), a typical Mediterranean fruit crop, is characterized by large adaptive potentialities to various ecological areas. To create a molecular characterization for 16 cultivars of Ficus, five  primer combinations were used to amplify these cultivar’s genomic DNA, producing a total of 292 legible bands (markers) were revealed, of which 281(96%) distinct polymorphic band patterns. The genetic diversity among 16 cultivars of fig ranged between 0.2124-0.7154. The lowest genetic distance was found between Qashe kani and Arzani where as the highest genetic distance was found between Qarani rash and Ribari. Cluster analysis by Using UPGMA method based on the similarity coefficient, cultivars were separated into seven major genetic clusters which were named as (F1, F2, F3, F4, F5, F6 and F7) with many sub-clusters. The analyzed data illustrates a good variability in the genetic pool of the local common fig making it a valuable source for incorporation into potential breeding programs for the region

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

Genetic Variation Assessment of Some Prunus Species Using Srap Markers

Genetic variation at molecular level was evaluated among three species of Prunus genus and a wild species, (P. arabica (wild), P. argentea (wild), P. dulcis (local), and  P. dulcis (wild ). The genetic variation assessment was carried out using SRAP molecular markers. The genetic similarity coefficient revealed the genetic relationship among the samples tested in which the highest genetic distance was between P. dulcis (local), and  P. dulcis (wild), and lowest genetic distance was between P. arabica (wild), P. argentea  (wild). The phylogenetic tree was obtained using UPGMA method depending on the total number of SRAP bands. There were two main groups in the dendrogram: the first one consists of two subgroup: P. arabica and P. argentea cluster together in one Subgroup and P. dulcis (wild) appear alone in this subgroup. P. dulcis (Local variety) appear in the second group alone

Bacteriological and Molecular Characterization of Extended Spectrum Β-Lactamases in Clinical Isolates of Klebsiella Pneumoniae Isolated From Kurdistan Region, Iraq.

A total of 275 clinical isolates of Klebsiella pneumoniae were collected from three general hospitals in Duhok, Erbil, and Sulymania, during the period September 2010 to June 2011. The Minimum Inhibitory Concentration (MIC) of these isolates was measured using the Gram-negative susceptibility card (GNC) of Phoenix system. Only 187 ESBL producing K. pneumoniae isolates were detected by this system. These isolates were confirmed as 100% ESBLs producers by the Double Disk Synergy Test (DDST). All 187 K. pneumoniae isolates were 100% resistant to ampicillin, cefazolin, cefepime, ceftriaxone, cefotaxime, cefuroxime, ceftazidime, and aztreonam. These isolates showed different percentages of resistance 81.8%, 68.5%, 65.8%, 52.4%, 50.3%, 34.2%, 25.2%, and 12.3% towards, ampicillin/sulbactam, gentamicin, trimethoprime-sulfamethoxazole, ciprofloxacin, piperacillin-tazobactam, amikacin, amoxicillin-clavulanate, and levofloxacin respectively.  Molecular characterization by PCR was employed using specific primers for three different ESBLs (TEM, SHV, and CTX-M). Results obtained revealed that SHV-type ESBLs were the most common ESBL occurring in 87% of the isolates with phenotypic evidence of ESBLs production. While those for TEM-type and CTX-M-type were 60% and 58% respectively

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

Evaluación de variantes genéticas y filogenia de la pera común (Pyrus communis L.) cultivada en la ciudad Duhok empleando AFLP como marcadores moleculares

Introduction: Genotyping and evaluation of genetic variation and polymorphic information content of the locally cultivated pear (Pyrus communis L.) might play an important role in building the genetic bank. These are also immensely important for present and future pear breeding program in the region. Methods: In the current study, AFLP markers have been employed to estimate the level of genetic diversity and to assess the phylogeny among theseven most popular pear cultivars in Duhok city. Results: Eight selective primer combinations generated a total of 653 AFLP fragments from which 445 (68.2%) fragments were polymorphic. The number of visible amplified products per primer combination were varied and ranged from 66 to 96 bands. The highestpercentage of polymorphism (78.4%) was observed by the primer pair P174/M182, while the lowest percentage of polymorphism (58.6%) was observed by the primer pair P174/M100. The highest PIC (0.85) was obtained with the primer combination P174/ M182, while, the lowest PIC (0.49) was obtained by the primer combination P174/M307. The genetic distance was ranged from 0.1348 (between Danimarki and Amreki cultivars) to 0.3131 (between Italy and Zaafaran2 cultivars). Based on the AFLP data, all the seven pear genotypes were successfully clustered into two separate clusters (C1 and C2) with an out-group of Itali cultivar. Conclusions: Overall, it can be concluded that there was high polymorphism among the studied genotypes. Also, it can be stated that the AFLP was a reliable and a powerful technique in genotyping and discriminating of respective pear cultivars

Identification of P53 Tumor Suppressor Gene Alterations in Human Glioma Using Sscp-Pcr Technique.

The P53 is the most commonly altered gene in cancers, which is located on chromosome 17p. Because of it is important role in the cell cycle it considered as the guardian of the genome that maintains genome integrity. In this study we analyzed DNA from 15 surgical specimens of human glioma for mutations in the p53 gene along exons 6-9 using single-strand conformation polymorphism analysis of polymerase chain reaction product (PCR-SSCP). The frequency of P53 gene mutation in gliomas examined was 26.6% (4 of 15). All 4 mutated cases were in exon 6 and no mutation was observed in exon 7,8 and 9. The results suggest the involvement of P53 gene mutation in Glioma and it might play an important role in the tumorigenesis process of patients with brain tumor in preliminary study in Duhok province

Molecular characterization of glucose-6-phosphate dehydrogenase deficient variants in Baghdad city - Iraq

Background: Although G6PD deficiency is the most common genetically determined blood disorder among Iraqis, its molecular basis has only recently been studied among the Kurds in North Iraq, while studies focusing on Arabs in other parts of Iraq are still absent. Methods: A total of 1810 apparently healthy adult male blood donors were randomly recruited from the national blood transfusion center in Baghdad. They were classified into G6PD deficient and non-deficient individuals based on the results of methemoglobin reduction test (MHRT), with confirmation of deficiency by subsequent enzyme assays. DNA from deficient individuals was studied using a polymerase chain reaction-Restriction fragment length polymorphism (PCR-RFLP) for four deficient molecular variants, namely G6PD Mediterranean (563 C®T), Chatham (1003 G®A), A- (202 G®A) and Aures (143 T®C). A subset of those with the Mediterranean variant, were further investigated for the 1311 (C®T) silent mutation. Results: G6PD deficiency was detected in 109 of the 1810 screened male individuals (6.0%). Among 101 G6PD deficient males molecularly studied, the Mediterranean mutation was detected in 75 cases (74.3%), G6PD Chatham in 5 cases (5.0%), G6PD A- in two cases (2.0%), and G6PD Aures in none. The 1311 silent mutation was detected in 48 out of the 51 G6PD deficient males with the Mediterranean variant studied (94.1%). Conclusions: Three polymorphic variants namely: the Mediterranean, Chatham and A-, constituted more than 80% of G6PD deficient variants among males in Baghdad. Iraq. This observation is to some extent comparable to othe