تنوع توالی سیتوکروم-b ژنوم میتوکندی در اسبهای کرد ایران

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم دامی، دانشکده کشاورزی و منابع طبیعی دانشگاه رازی، کرمانشاه، ایران.

2 موسسه تحقیقات علوم دامی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

چکیده

بررسی توالی نوکلئوتیدی ناحیه سیتوکروم-b ژنوم میتوکندری در درون جمعیت‌ها می‌تواند شاخص مناسبی از میزان تنوع موجود در جمعیت مورد مطالعه باشد. تحقیق اخیر به منظور بررسی تنوع ژنتیکی در اسب کرد و رابطه فیلوژنتیکی اسب کرد با سایر نژادهای اسب در دنیا با استفاده از ناحیه سیتوکروم-b انجام شد. بدین منظور از 30 رأس اسب نژاد کرد خون‌گیری بعمل آمد و DNA آن به روش نمکی استخراج گردید، سپس با استفاده از آغازگرهای اختصاصی ناحیه سیتوکروم-b به طول 1029جفت باز تکثیر و تمام نمونه‌ها بعد از خالص‌سازی توالی‌یابی شدند. قطعه مورد نظر پس از توالی‌یابی، با نرم‌افزار BioEdit  ویرایش و قطعة 882 جفت بازی از آن استخراج شد. نمونه‌ها با توالی مرجع اسب به شماره دسترسی (X79547) با استفاده از رویه Clustal-W همردیف شدند. تعداد هاپلوتیپ، جایگاههای نوکلئوتیدی متغیر، تنوع هاپلوتیپی و تنوع نوکلئوتیدی با استفاده از نرم افزار DNASP4.0 تعیین شد. درخت فیلوژنی با استفاده از نرم افزار MEGA6.1 به روش Neighbour-joining ترسیم شد. تعداد 7 هاپلوتایپ با 7 جایگاه نوکلئوتیدی متغیر شناسایی شد. هاپلوتایپهای بدست آمده همگی در هاپلوگروه K قرار گرفتند. میزان تنوع هاپلوتیپی و نوکلئوتیدی به ترتیب 001/0±784/0 و 001/0±00218/0 برآورد شد. ترکیب نوکلئوتیدها در توالی شاخص شامل 27/21% نوکلئوتید A، 77/32% نوکلئوتید C، 15/13% نوکلئوتید G و 87/26% نوکلئوتید  Tبود. بر اساس نتایج آنالیز فیلوژنی، اسب کرد از نظر ژنتیکی به نژادهایی از ژاپن، چین ، نژادی از لهستان و عربستان نزدیکتر است، که نشان دهنده تشابه ژنتیکی بیشتر اسب کرد با نژادهای آسیایی و تا حدودی اروپایی می‌باشد..

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Mitochondrial DNA Cytochrome-b variability in Iranian Kurdish horse

نویسندگان [English]

  • Hasan Jalilian Majd 1
  • Sheida Varkoohi 1
  • Hamid Reza Seyedabadi 2
1 Department of Animal Science, College of Agriculture and Natural Resources, Razi University, Kermanshah, Iran.
2 Animal Science Research Institute of IRAN (ASRI), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
چکیده [English]

Investigation of mitochondrial genome sequence of cytochrome-b region within population can be a good indicator for diversity in the studied population. A recent study was conducted to investigate the genetic diversity in Kurdish horses and phylogenetic relationship of between Kurdish horse with other horse breeds in the world using cytochrome-b region. Blood samples were collected from 30 Kurdish horses and total DNA was extracted by modified salting out method. Cytochrome-b sequences were amplified by primers pairs with 1029 bp length and then were sequenced after purifying. The sequences were trimmed to 882 bp using BioEdit software. The samples were aligned with the horse reference sequence with access number (X79547) using Clustal-W package. Haplotype and polymorphic site numbers, Haplotype and nucleotide diversity were estimated using DNASP4 software. Phylogenetic tree was constructed in MEGA7 software by neighbor joining method. 7 haplotypes with 7 polymorphic site were identified. Whole haplotypes were belonged to K haplogroup. Haplotype and nucleotide diversities were 0.784±0.001 and 0.00218±0.001, respectively. The compositional frequency of consensus sequences was including: A base, 21.27%; C base, 32.77%; G base, 13.15% and T base, 26.87%. According to the phylogenetic analysis, Kurdish horses were genetically more closely related to Japanese and Chinese breeds and one Polish and Saudi Arabian breeds which shows that Kurdish horse has genetic similarities with Asian and some European horse breeds.

کلیدواژه‌ها [English]

  • Kurdish horse
  • Genetic Diversity
  • Mitochondrial Genome
  • Cyt-b
  • Phylogeny

Extended Abstract

Introduction

Comparative sequence analysis of mitochondrial DNA (mtDNA) represents the standard molecular approach for species identification, with a particular emphasis on the cytochrome (cytb) gene. The most common molecular genetic approaches to species identification involve analysis of the mitochondrial DNA cytochrome b gene in nucleotide sequence of the cyt b gene contains species specific. Cytochromeb (cyt-b) has been considered as one of the most useful genes for phylogenetic work, and is probably the best-known mitochondrial gene with respect to structure and function of its protein product. Kurdish horse is one of the most valuable horse genetic resources in the Middle East. Despite the existence of a diverse range of different breeds on native horses in Asia, genetic research on some of these horse breeds, such as the Kurdish horse, is rare.

 

 

Therefore, Our main goals in the present study is to explore the utility of cyt-b to identify genetic structure and genetic variations in Kurdish horse and determination of phylogenetic relationships between Kurdish horses and other horse breed in the world.

 

Materials and Methods

Total DNA was extracted from the collected blood samples by modified salting out method. The cytochrome b was amplified by PCR and then sequenced using ABI PRISM BigDyeTM Terminator Cycle Sequencing Ready Reaction Kit. Consequently, the sequences were trimmed to 882 bp using BIOEDIT to become comparable with other reported cytochrome b sequences in GeneBank. Sequence alignment was performed using CLUSTALW package. Haplotype and nucleotide diversity were estimated using

DNASP5.10 and phylogenetic tree was constructed by Neighbour joining method.

 

Results

Based on results, 7 different haplotypes and 7 polymorphic sites were detected. The largest haplotype

group consisted of 12 individuals. Haplotype diversity and nucleotide diversity 0.784±0.001and 0.00218±0.001, respectively. Kurdish horse showed a high haplotype and low nucleotide diversity. The compositional frequency of consensus sequences for base A was the highest (27.21%) compared to other three nucleotides (C = 32.77%, T = 26.87% and G = 13.15%).

 

Conclusion

According to the phylogenetic analysis, Kurdish horses were genetically more closely related to Japanese (Hokkaido and Yakutia), Chinese (Lijiang and Sanhe), Polish and Arabic hors breeds, which indicates the greater genetic similarity of the Kurdish horse with Asian and to some extent European breeds.

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علوم دامی ایران
دوره 54، شماره 2
تیر 1402
صفحه 139-149

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  • تاریخ دریافت: 03 اردیبهشت 1401
  • تاریخ بازنگری: 02 مهر 1401
  • تاریخ پذیرش: 06 مهر 1401

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