دراسة التنوع الوراثي للجاموس العراقي باستخدام تقنيتي RAPD وSTR == Study of Genetic Diversity of Iraqi Buffalo by RAPD and STR
Author name:
فالح حسن حمد
Supervisor name:
اسعد يحيى عايد | طالب احمد جايد
General topic:
Agricultural sciences
Specific topic:
Zoology
Degree:
Master
University:
University Of Basrah - Faculty Of Agriculture
Language:
Arabic
University location:
Basrah
First pages:
31T1844 - p.pdf
Abstract:
The current study was conducted in the laboratory of Dr. TalibAhmed Jaayide Molecular Genetics / Faculty of Agriculture /University of Basrah. After collecting blood samples from the Iraqibuffalo in the provinces of Basrah (30 samples), Dhi Qar, Maysanand Najaf (25 samples each), bringing the total samples to 105samples. The study aimed to study the genetic diversity of the Iraqibuffalo through the use of tow techniques, first technique was, thePCR - RAPD technique and the second microsatellite technique(PCR - STR). After extracting the DNA samples, seven primers ofPCR - RAPD technique were used, included C01, C04, C06, C07,C09, C11 and C12. Primers of PCR - STR technique wereILSTS005, ILSTS029 and ILSTS072. After analyzing the results ofthe three technologies it can be summarized as follows : 1 - All primers of PCR - RAPD showed genetic polymorphisms,with the presence of many bands except the primer C11which was notamplified in all samples.2 - The size of primer C07 was 1969 bp, which was the biggestprimer size, however, the smallest primer size was C01(1606 bp).3 - Number of bands showed by PCR - RAPD primers were 28,42, 56, 53, 57 and 64 bands of C01, C04, C06, C07, C09,C12 primers respectively.4 - Primer C09 revealed the highest number of polymorphicbands in Basra and Dhi - Qar provinces (47 and 46 bandsrespectively). However, the primer C12 gave highest numberof polymorphic bands in Maysan (47 bands). The primer C01showed lowest number of polymorphic bands in all studiedprovinces (14 bands in Basra and Dhi - Qar provinces, 12bands in Maysan and 13 bands in Najaf province.5 - Shared bands between Basra and Dhi - Qar, Basra andMaysan, Basra and Najaf, Dhi - Qar and Maysan, Dhi - Qar andNajaf and Maysan and Najaf were 177, 151, 162, 146 and152 bands respectively. The high number of shared bandsreflected high genetic similarly among provinces.6 - Maysan province showed highest special bands of primerC07 (8 bands) followed by Dhi - Qar province (5 bands) whichis considered as fingerprint for these provinces even they showed very low frequencies.7 - Primer C04 revealed lowest genetic diversity measured by Shannon index in all studied areas (28.82 - 35.58%), however, all other primers showed higher genetic diversity (47.57 - 55.70%).8 - Similarity among provinces was higher than 70% to 95% for all PCR - RAPD primers, which indicates that all subpopulations of buffalo belong to one breed.9 - Primers C07 and C12 highlighted highest percent of mean gene diversity to population diversity (GST) with a value of 22.69 and 21.46 respectively.10 - All STR markers (ILSTS005, ILSTS029 and ILSTS072) have been amplified in all buffalo samples of all studied areas. Observed allele numbers were 20, 22 and 21 alleles for markers respectively with a total of 63 alleles. The marker ILSTS005 gave highest number of observed alleles in Dhi - Qar (21), Najaf (20) and Maysan (17). Whereas, the marker ILSTS029 showed less number of observed alleles at Maysan province (9 alleles). 11 - Allele frequencies ranged from 0.02 to 0.32 for all markers and provinces.12 - Shared alleles between Basra andDhi - Qar, Basra and Maysan, Basra and Najaf, Dhi - Qar and Maysan, Dhi - Qar andNajaf, Maysan and Najaf were 25, 27, 30, 22, 33 and 33alleles.13 - Special allele mean frequency of studied population was 0.048. While gene flow after adjusted for sample size of each province was 2.25 alleles. Range of missing allele frequency for all studied areas was 0.000 - 0.099.14 - Range of heterozygosity percent was 80 - 100%, whereas homozygosity% was 0 - 20%.15 - All Fis values of studied markers in all provinces were significant except the marker STR072 in Dhi - Qar province which was nonsignificant. This result reflected the absence of inbreeding in all areas and there was no significant deficiency in genetic variation.16 - Linkage disequilibrium test showed that tested markers are not located on one chromosome.17 - The study thus highlights the usefulness of heterologous PCR - RAPD and microsatellite markers to assess the genetic variability in buffalo. Also various diversity indices suggest sufficient genetic variability within Iraqi buffalo that can be utilized as initial guidelines for future breeding strategies and conservation.
