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تجزیهوتحلیل پروتئوم پیازچه درگونههای Allium stipitatum و A. scabriscapum ایرانی برای شناسایی پروتئینهای با بیان متفاوت | ||
| تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران | ||
| مقاله 6، دوره 29، شماره 2 - شماره پیاپی 58، اسفند 1400، صفحه 221-235 اصل مقاله (1.06 M) | ||
| نوع مقاله: مقاله علمی - پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/ijrfpbgr.2021.355432.1389 | ||
| نویسندگان | ||
| مهسا اسدی1؛ فرهاد نظریان فیروزآبادی* 2؛ محمد رضا نقوی3؛ احمد اسماعیلی4 | ||
| 1دانشجوی دکتری بیوتکنولوژی کشاورزی، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی و منابع طبیعی، دانشگاه لرستان | ||
| 2نویسنده مسئول مکاتبات، استاد، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی و منابع طبیعی، دانشگاه لرستان | ||
| 3استاد گروه زراعت و اصلاح نباتات، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، کرج | ||
| 4استاد گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی و منابع طبیعی، دانشگاه لرستان | ||
| چکیده | ||
| جنس Allium شامل بیش از 920 گونه در سراسر جهان است. ایران زیستگاه اصلی بسیاری از گونه های وحشی Allium میباشد. در این مطالعه از بافت پیازچه دو گونه A. stipitatum و A. scabriscapum از دو زیرجنسMelanocrommyum و Reticulatobulbosa با هدف ارزیابی و شناسایی الگوی بیانی پروتئینهای ذخیرهای حاصل از روش SDS-PAGE استفاده شد. برای شناسایی لکه های پروتئینی و مشاهده تغییرات بیانی دو گونه از الکتروفورز ژل دوبعدی و طیفسنجی جرمیMALDI TOF/TOF MS استفاده گردید. آنالیز ژل های حاصل از الکتروفورز دوبعدی منجر به شناسایی 138 لکه پروتئینی شد. نتایج حاصل بیانگر شباهت اندک میان ژلها و الگوی بیانی متفاوت در بین نمونه ها بود که بر این اساس تعداد کمی لکه مشابه در هر دو ژل حاصل شد. نتایج نشان داد در پروفایل بیانی پروتئین های بافت پیازچه این نمونهها تفاوتهای معنی داری وجود داشت که با استفاده از آزمون T تعداد 9 لکه پروتئینی دارای بیشترین تغییرات معنیدار در سطح احتمال 5% شناسایی شدند و از بین آنها 3 لکه متفاوت با استفاده از روش طیفسنجی جرمی برای شناسایی انتخاب شد. نتایج آزمایش طیفسنجی جرمی و تطبیق داده های حاصل با پایگاه دادههای NCBI و Uniprot و ابزارهای بیوانفورماتیکی مشخص کرد که لکه های مورد آزمایش با پروتئین های Maturase k، Agmatine coumaroyltransferase-1 و یک پروتئین با عملکرد نامشخص بنام Hypothetical Protein مطابقت داشت. یافته های این مطالعه نشان داد با توجه به اینکه نمونه های متعلق به دو گونه A.stipitatum و A. scabriscapum از یک جنس هستند اما الگوی بیانی پروتئینهای ذخیرهای آنها در بافت پیازچه بسیار متفاوت از یکدیگر است. | ||
| کلیدواژهها | ||
| آلیوم؛ آنالیز طیفسنجی جرمی؛ بیان ژن؛ بیوانفورماتیک | ||
| عنوان مقاله [English] | ||
| Scallion proteome analysis in Iranian A. stipitatum and A. scabriscapum species to identify proteins with different expression | ||
| نویسندگان [English] | ||
| M. Asadi1؛ F. Nazarian-Firouzabadi2؛ M. R. Naghavi3؛ A. Ismaili4 | ||
| 1PhD student Agricultural Biotechnology , Dept. Production Engineering and Plant Genetics, Faculty of Agriculture, Lorestan University, Khorramabad, IRAN | ||
| 2Corresponding Author, Prof. Department of Production Engineering and Plant Genetics, Faculty of Agriculture, Lorestan University, Khorramabad, I.R. Iran | ||
| 3Prof, Dept of Agronomy and Plant Breeding, College of Agriculture and Natural Resources University of Tehran, Karaj, I.R. Iran | ||
| 4Prof , Dept. Production Engineering and Plant Genetics, Faculty of Agriculture, Lorestan University, Khorramabad, IRAN | ||
| چکیده [English] | ||
| Allium includes more than 920 species worldwide. Iran is the main habitat of many wild Allium species. In this study, onion tissue of two species A. stipitatum and A. scabriscapum from two subspecies Melanocrommyum and Reticulatobulbosa were used to evaluate and identify the expression pattern of storage proteins obtained by the SDS-PAGE method. Two-dimensional gel electrophoresis and MALDI TOF/TOF MS mass spectrometry were used to identify protein spots and to observe the expression changes of the two species. Analysis of gels by two-dimensional electrophoresis revealed 138 protein spots. The results showed a slight similarity between the gels and a different expression pattern between the samples, based on which a small number of similar spots were obtained in both gels. The results showed that there were significant differences in the expression profile of onion tissue proteins of these samples. Using the T-test, 9 protein spots with the most significant changes were identified at the 5% probability level, and among them, 3 different spots were identified using Mass spectrometry was selected for identification. The results of mass spectrometry experiments and matching of the obtained data with NCBI and Uniprot databases and bioinformatics tools showed that the tested spots were consistent with Maturasek proteins, Agmatine coumaroyltransferase-1, and a protein with the unknown function called Hypothetical Protein. The findings of this study showed that since the samples belonging to A.stipitatum and A.scabriscapum are of the same genus, but the expression pattern of their stored proteins in onion tissue is very different from each other. | ||
| کلیدواژهها [English] | ||
| Allium, Mass spectrometry analysis, Gene expression, Bioinformatics | ||
| مراجع | ||
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