مطالعه پاسخ پروتئینی ریشه آفتابگردان به تنش خشکی با رهیافت پروتئومیکس

نوع مقاله : علمی پژوهشی

نویسندگان

1 موسسه تحقیقات، اصلاح و تهیه نهال و بذر، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

2 دانش‌آموخته کارشناسی‌ارشد زیست‌شناسی، دانشگاه پیام نور، تهران

3 استاد گروه زیست‌شناسی، دانشگاه پیام نور، تهران

چکیده

برای درک مکانیسم مولکولی تحمل به خشکی در ارتباط با ریشه آفتابگردان، الگوی پروتئوم ریشه دو لاین حساس و متحمل به خشکی در شرایط آبیاری محدود و مطلوب بررسی شد. با استفاده از الکتروفورز دو بعدی و مقایسه شدت نسبی لکه‌ها با آزمونt تعداد 12 لکه از 417 لکه پروتئینی در لاین حساس و 17 لکه از 467 لکه پروتئینی در لاین متحمل به طور معنی‌داری از تنش خشکی متأثر شد. لکه‌های پروتئینی با استفاده از طیف‌سنجی جرمی nano-LC MS/MS توسط جستجوگر مسکات با در نظر گرفتن حداقل 10% همپوشانی و امتیاز بالاتر از 80 در پایگاه NCBI شناسایی شدند. پروتئین‌های سیتوپلاسمی و هسته‌ای به ترتیب بیش از سایر پروتئین‌ها از محدودیت آب متأثر شدند. سه لکه پروتئینی انولاز، گلیسرآلدئید تری فسفات دهیدروژناز و چالکون سینتاز با بیان متمایز در هر دو لاین حساس و متحمل شناسایی شد. کاهش شدت نسبی انولاز در لاین متحمل دلالت بر آسیب متابولیکی بود که می‌تواند منجر به کاهش فرایندهای پایین دست در اثر محدودیت ناشی از خشکی باشد. در مقابل افزایش شدت نسبی گلیسرآلدئید تری‌فسفات دهیدروژناز و چالکون‌سینتاز در لاین متحمل نشان‌دهنده توانایی این لاین در سمیت‌زدایی/ترمیم آسیب-های ناشی از تخریب اکسیداتیو و دفاع ضداکسیداتیو در شرایط تنش بود. افزایش شدت نسبی پروتئین شوک‌دمایی، دی‌هیدروفلاونول ردوکتاز، لیپو-اکسیژناز لینولئات‌دانه، یوبیکوئیتین کربوکسیل‌ترمینال هیدرولاز و جی پروتئین در لاین متحمل نشان‌دهنده اهمیت فرایندهای دفاعی، حفاظتی و ترارسانی برای کاهش آسیب‌های ناشی از خشکی است.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

The study of protein response of sunflower root to drought stress using proteomics approach

نویسندگان [English]

  • Mehdi Ghaffari 1
  • Saadat Sharifi 2
  • Gholamreza Bakhshi Khaniki 3
1 Seed and Plant Improvement Institute Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran
2 Former M.Sc. of Biology, Payame Noor University, Tehran, Iran
3 Professor, Department of Biology, Payame Noor University, Tehran, Iran
چکیده [English]

In order to understand of the molecular mechanisms of drought tolerance in sunflower, proteomic pattern of roots in two drought sensitive and drought-tolerant lines were evaluated under limited and favorable water conditions. After 2DE and comparison of relative abundance of protein spots using t test, 12 of 417 protein spots in sensitive and 17 of 467 in tolerant line were affected by drought stress significantly. Following nano-LC MS/MS the protein spots were identified using Mascot search engine in NCBI protein database considering more than 10 % sequence coverage and score of above 80. Cytoplasmic and nuclear proteins were the most proteins which were affected by water deficiency. Three protein spots i.e. Enolase, Glyceraldehyde 3-phosphate dehydrogenase and Chalcone synthase were expressed differentially in these lines. Reduction of Enolase as a sign of metabolic impairment could be resulted in downstream process under drought stress. Increased expression of Glyceraldehyde 3-phosphate dehydrogenase and Chalcone synthase could have a role in detoxification/removal of oxidative destruction and antioxidant capability of the tolerant line. Increased level of heat shock protein, dihydroflavonol reductase, Seed linoleate 9S-lipoxygenase, Ubiquitin carboxyl-terminal hydrolase and G protein indicated crucial role of defensive, protective and transductive process in reduction of drought injuries.

کلیدواژه‌ها [English]

  • Two Dimensional Electrophoresis. Enolase
  • heat shock protein
  • Mass Spectrophotometry

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