شناسایی بیوانفورماتیکی و بررسی بیان miRNA های دخیل در نمو دانه ارقام گندم ایرانی با کیفیت نانوایی متفاوت

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش آموخته دکتری، گروه بیوتکنولوژی کشاورزی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت، ایران

2 استاد، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت، ایران

3 دانشیار، گروه تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران

4 استاد، گروه تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه زنجان، زنجان، ایران

5 استادیار، پژوهشکده فناوری‌های نوین زیستی، دانشگاه زنجان، زنجان، ایران

چکیده

کیفیت نانوایی گندم (Triticum aestivum L.) به­شدت وابسته به حضور و ترکیب پروتئین­های گلوتن در دانه است. در این مطالعه، کیفیت نانوایی چهار رقم گندم ایرانی (مروارید، سرداری، پارسی و سپاهان) بر اساس آنالیزهای فارینوگرافی و شیمیایی آرد و حجم ویژه نان به­ترتیب از بیش­ترین به کم­ترین کیفیت رتبه­بندی شد. به­منظور شناسایی و مقایسه miRNA های دخیل در نمو دانه بین گندم­های ایرانی با کیفیت نانوایی خوب و ضعیف، داده­های RNA-seq هفت رقم گندم (چهار رقم با کیفیت خوب و سه رقم با کیفیت ضعیف) در دو مرحله نموی دانه از پایگاه اطلاعاتی NCBI پیاده­سازی و آنالیز شدند. از miRNA های شناسایی شده، پنج miRNA حفاظت­شده انتخاب و الگوی بیان آن­ها با استفاده از روش Real Time PCR در دو مرحله نموی دانه (10 و 20 روز پس از گرده­افشانی) در چهار رقم گندم  ایرانی بررسی شد. نتایج نشان داد که الگوی بیان miR159a، miR164a، miR172a و miR827a طی نمو دانه در همه ارقام مورد بررسی روند افزایشی داشت، در حالی­که الگوی بیان miR396c روندی کاهشی نشان داد. miR164a و miR396c بیش­ترین و کم­ترین بیان نسبی را به­ترتیب در ارقام سپاهان و سرداری (به­ترتیب با کم­ترین و بیش­ترین وزن هزار دانه) نشان دادند که می­تواند در نتیجه نقش تنظیم­کنندگی منفی آن­ها بر اندازه دانه باشد. همچنین، با افزایش کیفیت نانوایی ارقام مورد بررسی، بیان نسبی miR172a و miR159a که به­ترتیب در سنتز نشاسته و پروتئین­های ذخیره­ای دانه دخیل هستند، کاهش و بیان miR827a که در فرایند انتقال مجدد نیتروژن از برگ به دانه دخیل است، افزایش یافت که نشان­دهنده آثار منفی miR172a و miR159a و مثبت miR827a بر کیفیت نانوایی ارقام گندم می­باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Bioinformatic identification and expression analysis of microRNAs involved in grain development of Iranian wheat cultivars with different bread-making quality

نویسندگان [English]

  • Reza Samimifard 1
  • Babak Rabiei 2
  • Bahram Maleki Zanjani 3
  • Jalal Saba 4
  • Abbas Bahari 5
1 Ph. D Graduate, Dept. of Agricultural Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran
2 Prof., Dept. of Plant Production and Genetic Engineering, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran
3 Assoc. Prof., Dept. of Plant Production and Genetic Engineering, Faculty of Agriculture, University of Zanjan, Zanjan, Iran
4 Prof., Dept. of Plant Production and Genetic Engineering, Faculty of Agriculture, University of Zanjan, Zanjan, Iran
5 Assist. Prof., Modern Biological Technologies Institute, University of Zanjan, Zanjan, Iran
چکیده [English]

The bread wheat (Triticum aestivum) quality is highly dependent on the presence and composition of the gluten proteins in the grain. In this research, bread-making quality of four Iranian bread wheat cultivars (Morvarid, Sardari, Parsi, and Sepahan) was ranked from the highest to lowest based on farinographic and chemical analyses of the wheat flour and bread specific volume. To identify and compare miRNAs involved in grain development between Iranian wheat cultivars with good and poor bread-making quality, RNA-seq data of seven wheat cultivars (four cultivars with good quality and three with poor) at two grain developmental stages were downloaded and analyzed from the NCBI database. Five conserved miRNAs were selected from the identified miRNAs and their expression pattern at two grain developmental stages (10 and 20 days after anthesis) were evaluated in four Iranian wheat cultivars using Real Time PCR. The results showed that the expression pattern of miR159a, miR164a, miR172a and miR827a during grain development in all studied cultivars had an increasing trend, while the expression pattern of miR396c showed a decreasing trend. miR164a and miR396c showed the highest and lowest relative expression in Sepahan and Morvarid (with the lowest and highest 1000-grain weight), respectively, which can be due to their negative regulatory roles on grain size. Also, with increasing the baking quality of the studied cultivars, the relative expression of miR159a and miR172a involved in the synthesis of starch and grain storage proteins, respectively, reduced and the expression of miR827a involved in the remobilization of nitrogen from leaves to grains, increased indicating negative effects of miR159a and miR172a and positive effect of miR827a on the bread-making quality of wheat cultivars.

کلیدواژه‌ها [English]

  • Farinograph analyses
  • Gluten proteins
  • miRNA expression pattern
  • qRT-PCR
  • RNA-seq
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