شناسایی و بررسی الگوی بیان برخی ژن‌های دخیل در تحمل به تنش خشکی در عدس (Lens culinaris)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش آموخته کارشناسی ارشد، پردیس کشاورزی و منابع طبیعی دانشگاه تهران

2 استاد، گروه زراعت و اصلاح نباتات، دانشکده علوم و مهندسی کشاورزی، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، کرج، ایران

3 دانشیار، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج، ایران

4 استادیار، دانشگاه تربیت مدرس، تهران، ایران

5 دکتری ژنتیک و به‌نژادی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج، ایران.

چکیده

هدف: به‌‌ دلیل کمبود آب و تنش خشکی آخر فصل در کشت دیم عدس در ایران، مطالعه حاضر با هدف شناسایی ژن‌‌های دخیل در تحمل به تنش خشکی و بررسی تغییرات آن‌‌ها به ‌منظور شناسایی نقش آن‌ها در این فرایند، در ژنوتیپهای حساس و متحمل عدس، انجام شد.
مواد و روش‌ها: به ‌منظور شناسایی توالی توافقی ناحیه رمزکننده ژن‌های انتخابی دخیل در تحمل به تنش خشکی گیاه عدس، ابتدا توالی‌‌های قطعات بیانی و خوانش ژن‌‌هایAQUAPIP2 ،FBA، OEE2، WRKY1 و XTH با استفاده از توالی‌‌های ارتولوگ در سایر گیاهان هم‌‌خانواده عدس شناسایی شدند. در پژوهش حاضر، به ‌منظور القای تنش خشکی، از آبیاری 60،90 و 30 درصد ظرفیت زراعی به ترتیب برای شاهد، تنش متوسط و شدید استفاده ‌شد. RNA گیاه عدس، از بوته‌‌های تحت تنش و شاهد استخراج، cDNA آن‌‌ها تهیه و بیان ژن‌های شناسایی شده، توسط تکنیک qRT-PCR، سنجیده‌ شد. برای درک بهتر پاسخ، تاثیر تنش خشکی بر برخی صفات مورفولوژیک، فیزیولوژیک و بیوشیمیایی گیاه عدس، ارزیابی شد.
یافته‌‌ها: با افزایش شدت تنش خشکی، ارتفاع بوته، وزن تر اندام هوایی و ریشه، وزن خشک اندام هوایی و ریشه، محتوای نسبی آب برگ، شاخص سطح برگ، کلروفیل و پروتئین برگ، کاهش و تراکم کرک برگ، میزان نشت الکترولیتی، پرولین و کربوهیدرات، محتوای مالون‌دی‌آلدهید و غلظت هیدروژن پراکسید، افزایش یافتند. نتایج حاصل از تجزیه و تحلیل داده‌های بیان نسبی ژن نشان داد که با افزایش شدت تنش، بیان نسبی ژن‌هایAQUAPIP2 ، FBA و WRKY1، افزایش معنی‌دار و بیان XTH و OEE2 کاهش معنی‌دار یافت.
نتیجه‌گیری: افزایش شدت تنش خشکی، منجر به افزایش بیان نسبی ژن‌های AQUAPIP2، FBA و WRKY1 شد. افزایش محتوای نسبی آب برگ و فعالیت آنزیم‌های آنتی‌اکسیدان وکاهش غلظت هیدروژن پراکسید، تحت تاثیر افزایش بیان AQUAPIP2 و WRKY1 بود. افزایش بیان نسبی FBA منجر به افزایش غلظت کربوهیدرات شد. نتایج نشان می‌دهد که این سه ژن دخیل در تحمل به تنش خشکی در عدس بودند.

کلیدواژه‌ها


عنوان مقاله [English]

Identification and study of changes in expression pattern of some genes involved in drought tolerance in Lentil (Lens culinaris)

نویسندگان [English]

  • Sahar Mobasser 1
  • Mohammad Reza Bihamta 2
  • Ali Reza Abbasi 3
  • Sajad Rashidi Monfared 4
  • Abdolrahman Rasoulnia 5
1 M.Sc. Graduate of Genetic and plant breeding, Department of agronomy and plant breeding, Faculty of Sciences and Agricultural Engineering, Tehran University
2 , Department of Agronomy & Plant Breeding, Faculty of Agronomy Sciences, College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran.
3 Associate Professor, Department of Agronomy & Plant breeding, Faculty of Agronomy Sciences, College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
4 Assistant Professor, Department of Agricultural Biotechnology, Tarbiat Modares University, Tehran, Iran
5 PhD of plant breeding, Department of Agronomy & Plant breeding, Faculty of Agronomy Sciences, College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
چکیده [English]

Objectives
The research was conducted to study changes in expression pattern of some genes involved in drought tolerance due to the problem of terminal drought stress in rainfed lentil agriculture in Iran.
Material and methods
In order to identify the consensus sequence of the coding sequence (CDS) of selected lentil genes; Aquaporin PIP2, Fructose bisphosphate aldolase, Oxygen-evolving enhancer protein2, WRKY1 and Xyloglucan endotransglucosylase/hydrolase, expressed sequence tag (EST) and reads were identified through similar ortholog sequences in other leguminosae plants. 90, 60 and 30 percent of field capacity were considered as control, medium and severe stress respectively. RNA was extracted from both control and severe drought stress treated plants leaf samples, cDNA was synthesized from RNA samples. Expression profiles of the genes, were determined via real time PCR technique. To represent the effect of drought stress on lentil plant, some morphological, physiological and biochemical characteristics were measured.
Results
Characteristics like plant height, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight, relative water content, leaf area index, chlorophyll and protein content decreased due to drought stress, leaf trichome density, electrolyte leakage index, proline and carbohydrate content, malondialdehyde and hydrogen peroxide increased in drought stress. Analysis of real time PCR demonstrated expression of AQUAPIP2، FBA and WRKY1 genes were increased and expression of XTH and OEE2 genes were decreased under severe drought stress condition.
Conclusions
The results show that AQUAPIP2, FBA and WRKY1 genes are involved in drought tolerance. Due to higher expression of AQUAPIP2 and WRKY1 genes in the tolerant genotype, reduction of RWC in this genotype is less. Less increase in hydrogen peroxide in the tolerant genotype, partly influenced by higher expression of AquaPIP2 gene and higher activity of antioxidant enzymes, which was influenced by higher expression of AquaPIP2 gene in tolerant genotype than susceptible genotype. The increase in carbohydrate content in both genotypes was partially affected by FBA gene expression.

کلیدواژه‌ها [English]

  • Key words: Drought stress
  • lentil
  • ortholog sequences
  • susceptible genotype tolerant genotype
علیزاده بهزاد، مصطفوی خدا داد، زمانیان محمد (1395) بررسی میزان تحمل به خشکی در ارقام شبدر برسیم و ایرانی. مجله زراعت و اصلاح نباتات 12، 76-67.
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