توسعه نشانگرهای ریزماهواره EST-SSR مرتبط با تحمل به تنش خشکی در عدس (Lens culinaris)

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی دکتری گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران.

2 دانشیار گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران.

3 استاد گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران.

4 استادیار گروه زیست‌شناسی، دانشکده علوم پایه، دانشگاه رازی، کرمانشاه، ایران.

5 دانشیار گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران.

چکیده

عدس یکی از لگوم‌های دانه‌ای مهم از نظر غذایی (به عنوان منبع غنی از پروتئین) و صنعتی (مثل صنعت بیوپلیمر) است و عملکرد پایین این گیاه در ایران نسبت به متوسط جهانی متأثر از تنش‌های محیطی به‌ویژه خشکی است. شناسایی نشانگرهای مولکولی مرتبط با ژن‌های دخیل در مقاومت به خشکی در ژنوم گیاه می‌تواند به‌نژادگران را در انجام برنامه‌های اصلاحی گیاهان مقاوم به خشکی کمک نماید. در این مطالعه هدف شناسایی نشانگرهای EST-SSR پیوسته با ژن‌های پاسخ‌دهنده به تنش خشکی بود که امید می‌رود که از این اطلاعات به منظور شناسایی ژنوتیپ‌های مقاوم به خشکی در برنامه‌های به‌نژادی بهره برد. جهت اعمال تنش از پلی‌اتیلن‌گلیکول استفاده شد و پس از پایان تنش نمونه‌گیری از بافت برگ انجام شد. سپس RNA کل استخراج و کتابخانه‌های cDNA مورد توالی‌یابی قرار گرفت. نتایج نشان داد که تعداد 10547 (16 درصد) از یونی‌ژن‌ها حداقل یک توالیEST-SSR را دارا بودند و حدود 5/27 درصد از این یونی‌ژن‌ها مستندسازی و در نهایت برای آنها رابطه هستی‌شناسی تعیین شد. بیشترین نشانگرها به ترتیب مربوط به تکرار‌های 1، 3 و 2 نوکلئوتیدی (با فراوانی 03/46، 25/37 و18/15 درصد) بودند. نتایج مستند سازی کارکردی این یونی‌ژن‌ها نشان داد که بیشترین تعداد EST-SSRها به ترتیب به زیر گروه اتصال با 872، زیرگروه فعالیت کاتالیتیکی با 806، فرآیند متابولیکی با 755 و بخش‌های سلول با 651 یونی‌ژن اختصاص داشت. نتایج نشان داد که ژن‌های دارای این نشانگرها در اعمال حیاتی مهمی دخیل هستند و ابزار مناسبی برای مطالعۀ ژن‌های دخیل در تحمل به تنش‌ها از جمله تنش خشکی هستند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Development of EST-SSR Microsattelite Markers Related to Drought Tolerance in Lentile (lens culinaris)

نویسندگان [English]

  • Seyedeh Zahra Hosseini 1
  • Ahmad Ismaili 2
  • Farhad Nazarian Firouzabadi 3
  • Hossein Fallahi 4
  • Abdolhossein Rezaeinejad 5
1 Ph.D. Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
2 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
3 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
4 Assistant Professor, Department of Biology, Faculty of Basic Sciences, Razi University, Kermanshah, Iran.
5 Associate Professor, Department of Horticultural Sciences, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
چکیده [English]

Lentil (Lens culinaris) is one of the important grain legumes in feeding (as protein-reach food) and industry (such as biopolymer industry) and the problem of lower yield of this plant in Iran rather than average global yield is affected by exposure of plant to environmental stresses especially drought. Identification of molecular markers that closely linked to drought resistant genes help to implementation of breeding programs aimed at the production of drought tolerant plants. The gol of this study was identification of EST-SSR markers which closely linked to the genes involved in drought resistance and use of these information in identification of drought resistant genotypes in breeding programs. PEG was used for stress treatment, and after conduction of treatments, leaf samples were collected. Total RNA was extracted and cDNA libraries were sequenced. Results showed that 10546 (16%) of uni-genes contained at least one EST-SSR and about 27.5% of these sequences were annotated. Among different SSR motif-classes, tri-nucleotide repeats (46.03%) were the most abundant followed by mono-nucleotide repeats (37.25%) and di-nucleotide repeats (15.18%). The results of the functional annotation of these sequences, showed that the highest number of EST-SSRs were belonged to subgroups of binding (872), catalytic activity (806), metabolic processes (755), and cell components (651), respectively.The results showed that genes associated with these markers, involved in important biological functions and are an appropriate tool for study the genes involved in tolerance to stresses including drought stress.

کلیدواژه‌ها [English]

  • Lentil
  • Marker
  • EST-SSR
  • Drought stress

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