Microsatillate based Parentage Verification in Crossbred Sheep Herds
محورهای موضوعی : Camelک. کسرایی 1 , س.ع. رافت 2 , آ. جوانمرد 3 , ج. شجاع 4 , م. عبدالسعید 5 , م. خان سفید 6
1 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
2 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
3 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
4 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
5 - Department of Animal Bioscience, Faculty of Veterinary Science, University of Sydney, Sydney, Australia
6 - Department of Environment and Primary Industries, AgriBio, Ring Road, Bundoora, VIC, 3083, Australia
کلید واژه: Sheep, Microsatellite, parentage control, pedigree errors,
چکیده مقاله :
Parentage testing is an important tool in farm animals for genetically determining the accuracy of pedigree information. The objective of the current study was to implication of multiplexing 14 microsatellite markers for routine parentage testings. The twenty-four lambs were crossbred of Ghazel × Baluchi, Ghazel × Baluchi × Merinos, and Baluchi × Moghani × Merinos breeds. The genomic DNA was extracted from the whole blood samples and genotyped using fragment analysis method. The highest and the reproducible multiplex group appeared by grouping ILSTS0049, MCM512, BMC1009, BM148 and CSSM032 loci in 5-plex reaction. The markers were first evaluated based on the number of alleles, allelic frequency, polymorphism information content (PIC), expected heterozygosity (HE), observed heterozygosity (HO) and the individual exclusion probability using popgene and cervus software. The average heterozygosity, polymorphism index content (PIC) and number of alleles per loci were 0.60, 0.58 and 4.93, respectively. The total exclusion probability of 14 microsatellite loci was 0.9999 in the population by compatibility according to the Mendelian fashion. The pedigree was considered incorrect in one out of all the evaluated progeny, as the genotype of that progeny did not match to any of its parents. The results of our study suggest the multiplex microsatellite panel a fast, robust, reliable, and economically efficient tool to verify the parentage and hence it can be used in the routine parentage testing in sheep.
در حیوانات مزرعه، آزمون انساب از ابزارهای ژنتیکی کلیدی، برای تعیین دقت و صحت اطلاعات شجره ثبت شده می‌باشد. هدف از پژوهش حاضر، ارزیابی و استفاده از سیستم تکثیر همزمان (گروهی) 14 جایگاه میکروساتلایت برای انجام یک آزمون معمول شناسایی والدین می‌باشد. بدین منظور، 24 نتاج آمیخته از نژادهای گوسفندان قزل × بلوچی، بلوچی × قزل × مرینوس و بلوچی × مغانی × مرینوس و والدین نسبت داده شده به آنها برای ادامه پژوهش انتخاب شدند. استخراج DNA ژنومی با استفاده از نمونه‌های خون کامل و از طریق الکترورز کاپیلاری برای جایگاه‌های انتخاب شده، تعیین ژنوتیپ شد. بالاترین سیستم مولتیپلکس طراحی شده متعلق به گروه جایگاه‌های میکروساتلایت پنج گانهILSTS0049 ، MCM512، BMC1009، BM148 و CSSM032 بود که نتایج شارپ و تکرارپذیری را تولید می‌کرد. کارآیی نشانگرهای انتخاب شده در این مطالعه در گام اول با استفاده از شاخص‌های همچون، تعداد الل مشاهده شده، فراوانی اللی، محتوای اطلاعات چند‌شکلی، هتروزیگوتی مورد مشاهده و مورد انتظار و احتمال حذف یک فرد از شجره با استفاده از نرم افزارهای پاپ ژن و CERVUS ارزیابی شد. ارزش‌های مشاهده شده برای سه شاخص میانگین هتروزیگوتی، محتوای اطلاعات چند‌شکلی و تعداد الل‌های مشاهده شده در کل این چهارده جایگاه به ترتیب 60/0، 58/0 و 93/4 بود. بر اساس الگوی توارثی مندلی جایگاه‌های مورد مطالعه، قدرت و دقت شاخص احتمال حذف یک فرد از شجره در حدود 999/0 برآورد گردید. نتایج این پژوهش به روشنی نشان داده که یکی از نتایج منتسب به والدین شناخته شده بر اساس مقایسه الگوهای باندی و ژنوتیپ والدین و این فرد تعیین ژنوتیپ شده ثبت اشتباه شده است (خطای شجره). در نهایت استفاده از سیستم تکثیر گروهی جایگاه‌ها و سیستم الکترورز مویینه (کاپیلاری) می‌تواند روشی دقیق، سریع و مقرون به صرفه برای تأیید والدین و شناسایی نتاج آنها برای انجام آزمون تأیید والدین در گله‌های مردمی گوسفند می‌باشد.
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