Genetic Variability Of Sahiwal And Cholistani Cattle Breeds Of Pakistan Usin Mitochondrial D-Loop Sequences
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: Pakistan is rich in cattle genetic resources. The phenotypic and genetic diversity of animal breeds in Pakistan is very vast. Efforts to manage and utilize these genetic resources efficiently are lacking due to lack of both awareness and weakness of Government institutions. The genetic data of dairy cattle breeds (Sahiwal and Cholistani) is not yet been studied for their genetic identification, conservation and to find the genetic diversity among them and it needs to be established. For this study the blood samples( 25 samples from each breed) were collected from their home tracts and livestock farms. Unrelated animals with typical phenotypic features known for Sahiwal and Cholistani cattle breeds were selected from their breeding areas and Government livestock farms. Blood samples from true representative individuals of Sahiwal breed were collected from Research Centre for the Conservation of Sahiwal Cattle (RCCSC), Jahangirabad, Khanewal, Semen Production Unit (SPU) Qadirabad and Barani Livestock Production & Research Institute (BLPRI), Kherimurat District Attock. Cholistani cattle samples were collected from Govt. Livestock Farm, Jugaitpeer, Bahawalpur. Sampling from siblings was avoided to minimize inbreed samples as it results in depleting of gene pool along with causing inbreeding depression.DNA was extracted and quantified with the standard protocol in Molecular Biology and Genomics Laboratory of Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore. Specific primers were designed by using special software i.e. Primer Fox for mitochondrial D-loop region from NCBI accession no. NC_006853.1.Primers optimization was done after primer designing and afterwards, PCR amplification was performed. Then sequencing of target fragments was carried out using Prism ABI 3130L sequencer and Analyser.Sequences were alligned with the help of software blast2sequence and SNPs were detected. It was found that ratio of transition mutation was higher than transversions i.e. 41 transition and 10 transversions. Sequences were analyzed and compared with already reported sequence of Mitochondrial DNA of Bosindicuss, Bostaurus, Bubalusbubalis, Canis lupus familiaris, Caprahircus, Equuscaballusisolate, Ovisaries and Cameliusdromedaries sequencesavailable at NCBI. Single Nucleotide Polymorphisms (SNPs) were then detected. A phylogenetic tree constructed using MEGA 5.1 software revealed that Pakistani, European and Asian cattle are genetically same but different from Buffalo.This work is very helpful about breed characterization of two cattle breeds (Sahiwal and Cholistani) and developing understanding about genetic architecture of cattle breeds as present study conclude that 52 SNPs were present in Sahiwal and Cholistani breed of Pakistan.
Availability: Items available for loan: UVAS Library [ Call number: 1498,T] (1).
Assessment Of Genetic Diversity In Balochi And Rakhshani Sheep Breeds Of Balochistan Using Microsatellite Dna
Material type: Book ; Format:
Publisher: 2011 Dissertation note: Pakistan being agriculture based country has a great potential in livestock sector, it plays an important role in the economy of the country. Pakistan is rich in Animal Genetics Resource (AnGR) and has various breeds of sheep but lacking genetic data of these breeds which need to established data for their genetic identification. Customarily, classification of breed was based on phenotypic traits. In some cases, recent genetic studies have found differences in the structure proposed. Molecular characterization is a prevailing tool to consider the genetic variation existed within and among breeds. Characterization and evaluation of genetic differences among these breeds is necessary for their effective and meaningful improvement and conservation. The advent of molecular techniques has led to an increase in the studies that focus on the genetic characterization of domestic breeds using genetic markers. Due to their reliability and availability, the microsatellites have become preferred method for the genome mapping. Microsatellites or STRs are the 2-6 nucleotide tandem repeats present in both coding and non coding regions of both prokaryotes and eukaryotes. Microsatellites are powerful tools in genome mapping, forensic DNA studies, paternity testing, population genetics and conservation/ management of biological resources.
The present study was conducted on the molecular diversity analysis of two sheep breeds Balochi and Rakhshani of Balochistan using 11 FAO recommended microsatellites markers. Blood samples of unrelated true representative animals of sheep breeds were selected from their breeding tracts and from different Government Livestock Farms in Balochistan province. DNA was extracted with the standard protocol and amplification of DNA done with selected markers in Molecular Biology and Genomics Laboratory in the Institute of Biochemistry and Biotechnology. PCR products were examined on non denaturing Polyacralamide Gel Electrophoresis (PAGE). Genotyping results vanalyzed through the software POPGENE VERSION 1.31 and "POWER STATE" for calculating the observed and expected number of alleles, expected and observed heterozygosity, homozygosity, F-statistics (FST, FIT, FIS), Polymorphic Information Content (PIC), matching probability power of discrimination and power of exclusion. This work provided the genetic data which is useful in breed identification and making effective breeding policies and conservational activities in future according to FAO global Farm Animal Genetic resource data.
Average observed heterozygosity, average observed homozygosity, observed number of allels (na) and expected number of alleles for all loci and population in this study were 0.6055, 0.3945, 6.3636 and 4.2805 respectively. Almost all of the microsatellite markers showed significant variations in both breeds of sheep.
This work provided the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in future according to FAO global Farm Animal Genetic resource data. Moreover this study can become the basis for further research investigations in sheep breeds in Balochistan and Pakistan.
Availability: Items available for loan: UVAS Library [ Call number: 1516,T] (1).
Develoopment Of A Reliable Microsatellites Maarkers Panel For Parentage Analysis In Cattle Breeds Of Pakistan and Its Validatio Through Cytochrome B Gene Sequencing
Material type: Book ; Format:
Publisher: 2013 Dissertation note: Pakistan posseses enormous Animal Genetic Resource (AnGR) with 36.9 millions of cattle population. The data on genetic fabric of these breed is yet to be documented for their genetic characterization and identification. This work reports first country wide microsatellite markers and cytochrome b gene based genetic characterization of 10 famous cattle breeds of Pakistan. A total of 352 blood samples from unrelated and phenotypically representative of ten native cattle breeds including Bos indicus; Sahiwal, Cholistani, Red Sindhi, Tharparker, Dhanni, Dajal, Lohai, Bhagnari, Achai and Bos indicus x Bos taurus; Nari Master, and an exotic Bos taurus; Holstein Friesian breeds were collected from their respective home tracts, institutional herds and private livestock farms located throughtout the country. These samples were subject to DNA extraction using inorganic method caliberated to same concentration in Molecular Biology and Genomics Laboratory of the Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore Pakistan. A total of 21 microsatellite markers recommended by the programme for the global management of genetic resources (MoDAD) for breed characterization of Food and Agriculture Organization (FAO) of the United Nations and International Society for Animal Genetics (ISAG) were applied. Multiplex PCR were optimized for amplification and were genotyped using ABI Genetic Analyzer 3130 xl using LIZ as size standard. Genotyping results were analyzed using POPGENE and Arlequin ver 3.5 software. The observed and effective number of alleles ranged from 10 (INRA32) to 43 (TGLA126) and 2.3574 (CSSM66) to 15.0019 (BM6526) respectively in all breeds? The observed and expected heterozygosity estimates ranged from 0.0638 (INRA32) to 0.7101 (BM2113) and 0.6510 (INRA32) to 0.9347 (BM6526) respectively in the experimental samples. Mean values for observed and expected heterozygosity was 0.4943 ± 0.1647 and 0.8164 ± 0.0930 respectively. Mean values for Fis, Fit and Fst in all cattle breeds were calculated as 0.2819, 0.3864 and 0.1456 respectively. Average polymorphic information content (PIC) of all microsatellite loci was 0.81 indicating a high degree of informativeness of all microsatellite markers used. It implies that the same set of markers is equally good and could reliably be used for parentage confirmation in Pakistani cattle breeds. The data produced, also showed least degree of genetic difference between Red Sindhi and Tharparker breeds. This may due to mixing of the two breeds for being in close proximity of their home tracts.
Fragment mitochondrial cytochrome b gene was also amplified using specific primers through PCR of 130 individuals representing all selected breeds and sequencing was done using ABI Genetic Analyzer 3130 xl. The sequences were aligned and analyzed with CodonCode Alligner 4.0.4 software. The analysis revealed highly degree of sequence conservation in all the Pakistani cattle while documenting changes in only 9 nucleotides from 26 individuals whereas multiple nucleotide changes in 5 locations were shown by more than one individual in the data presented. One polymorphic site was found in nucleotide 318 (T?C) in several breeds of indicine cattle while 2 Lohani and 5 Nari Master individuals showed nucleotide changes specific to taurine cattle. Of all the changes found, only three of them caused changes in the amino acid sequence. The UPGMA tree using MEGA 5.1 showed a clear differentiation between taurine and indicine cattle, except for Nari Master Pakistani cattle showing mitochondrial taurine sequences because it's a cross between Bhagnari (Bos indicus) and Australian Draught Master (Bos taurrus). The estimates of divergence among breeds were also low for most breed pairs, except for Nari Master and Dhanni whereas the overall divergence within Bos indicus or within Bos taurus were also very low (0.002 and 0.003, respectively) but the differences between Bos indicus and Bos taurus were significantly higher (0.014) as should be the case.
These results of microsatellite markers have produced a set of information that can be recommended as a reliable marker panel for studies on genetic diversity analysis, parentage confirmation. The cytochrome b data on the other hand not only substantiated genetic diversity analyses but it also proved to be equally good for comparative Phylogenetic analysis of Pakistani cattle breeds and exotic breeds. This work provides most authenticated data and adds a great deal, to already existing information on Pakistani AnGR. This information coupled with prospective data using next generation genetic technologies will assist designing breed improvement focused breeding policies and conservation activities in future.
Availability: Items available for loan: UVAS Library [ Call number: 1597,T] (1).