شناسایی نشانگرهای چندشکل ریزماهواره در چغندرقند

نوع مقاله: کامل علمی - پژوهشی

نویسندگان

1 دانشگاه بوعلی سینا

2 دانش آموخته کارشناسی ارشد دانشگاه بوعلی سینا

3 عضو هیئت علمی-دانشگاه بوعلی سینا

چکیده

به منظور شناسایی نشانگرهای SSR و EST-SSR در چغندر قند، مجموعا 2884 توالی EST و همچنین توالی ژن FLC-Like، یکی از ژن­های کلیدی کنترل کننده گلدهی در چغندر، از پایگاه NCBI استخراج شد. بعد از انجام سرهمبندی توالی­ها و ساخت توالی­های غیر تکراری، همه کانتیگ­ها به منظور شناسایی SSRsهای دو تا شش نوکلئوتیدی مورد جستجو قرار گرفتند. سپس طراحی آغازگرهای اختصاصی از نواحی حفاظت شده اطراف این SSRها انجام شد. تعداد 300 توالی معادل 14 درصد از کل توالی­های EST دارای ریزماهواره بودند و به طور متوسط نواحی بیان شونده در چغندرقند به ازای هر 4/6 کیلو باز دارای یک ریزماهواره بوده و فراوانترین آن­ها تکرارهای سه تایی  (36/33) بودند.  تعدا 15 جفت آغازگر اختصاصی طراحی شده بر روی 12 رقم و لاین چغندر با آزمون PCR بررسی شد و شش نشانگر چندشکل شناسایی شد که چهار مورد از آن­ها از توالی­های ژن­های کنترل کننده گلدهی هستند. این نشانگرها قادر به تولید دو تا چهار الل در هر مکان بوده و محتوای اطلاعات چندشکلی (PIC) آن­ها بین 0/4 تا 0/66 بود. این اطلاعات EST-SSR و نشانگرهای جدید SSR شناسایی شده، منابع مهمی برای مطالعات ژنتیکی، تاکسونومی، اصلاح مولکولی، شناسایی ارقام  در مطالعات آینده بر روی چغندر قند است.

کلیدواژه‌ها


عنوان مقاله [English]

Development of polymorphic microsatellite markers for sugar beet (Beta vulgaris L.)

نویسنده [English]

  • Asghar Mirzaie-asl 1
1 Bu-Ali Sina University
چکیده [English]

A total of 2884 Expressed Sequence Tags (EST) and genomic sequence of FLC-Like gene, a major gene controlling the flowering time in sugar beet, were extracted from NCBI database. After sequence alignment and construction of unrepeated primers, all contigs were surveyed for identification of 2-6 bp SSRs. Then, specific primers were designed from the conserved regions around the SSRs. Three hundred consensus sequences (14% of all EST sequences) contained SSRs with an average of 4.6 kb of expressed region of genome for one SSR. Trinucleotide repeats were the most frequent (36.33%) SSRs. Among the 15 specific primer pairs tested with PCR on 12 beet accessions, cultivars, and lines, six polymorph markers were identified four of which were the sequences of the flowering genes. These markers were able to generate 2-4 alleles per locus with minimum and maximum polymorphic information content (PIC) of 0.4 and 0.66, respectively. This EST-SSRs dataset and new identified SSR markers may be used as important sources for genetic and taxonomic studies, molecular breeding and identification of sugar beet varieties in the future studies.

کلیدواژه‌ها [English]

  • beta vulgaris
  • EST
  • flowering genes
  • SSR markers
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