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بررسی بیان ژن‌‌های کلیدی در مسیر بیوسنتز پولگن درگیاه کاکوتی ایرانی (Ziziphora persica Bunge) تحت تنش خشکی

نوع مقاله : علمی پژوهشی - اصلاح نباتات

نویسندگان

1 دانشجوی دکترا مهندسی تولید و ژنتیک گیاهی، گروه تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران

2 استادیار، گروه تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران

3 آزمایشگاه مرکزی منابع گیاهی انجمن باغبانی، اکادمی علوم چین، پکن، چین

چکیده

امروزه مبحث گیاهان دارویی و متابولیت‌های ثانویه به‌طور قابل توجهی مورد استقبال پژوهشگران قرار گرفته است. کاکوتی ایرانی (Ziziphora persica Bunge) از جمله گیاهان دارویی مطرح متعلق به خانواده lamiaceae می‌باشد که دارای اسانسی با ماده مؤثره پولگن (pulegone) فراوان است. خشکی یکی از عوامل محدود‌کننده رشد در سرتاسر جهان و شایع‌ترین تنش محیطی می‌باشد و یکی از مهم‌ترین عوامل محدود‌کننده تولید محصولات کشاورزی در سراسر دنیاست. مطالعات انجام‌شده بیانگر وجود تأثیر عمده این تنش بر میزان رشد، مواد مؤثره و هم‌چنین تاثیر قابل توجه بر میزان بیان ژن‌های دخیل در سنتز ماده مؤثره گیاهان می‌باشد. این مطالعه برای ارزیابی اثر احتمالی تنش خشکی بر ماده مؤثره پولگن در کاکوتی ایرانی انجام گرفت. از آنجایی که تحمل به خشکی یک صفت ساده ژنتیکی نبوده بلکه صفتی کمی و پیچیده با جنبه‌های مختلف می‌باشد، به منظور بررسی اثر تنش خشکی (30 درصد ظرفیت مزرعه) میزان پرولین و محتوای نسبی آب در شرایط تنش با شرایط کنترل مقایسه گردید. نتایج نشان داد سطح انباشت پرولین در شرایط تنش نسبت به شرایط کنترل به‌طور قابل توجهی افزایش می‌یابد که نشان از فعال شدن مکانیسم‌های تحمل در گیاه دارد. در این پژوهش اثر کمبود آّب (30 درصد ظرفیت مزرعه) بر میزان بیان 4 ژن دخیل در مسیر بیوسنتز پولگن مورد مطالعه قرار گرفت. افزایش سطوح بیان ژن‌های لیمونن سنتتاز، لیمونن هیدروکسیلاز، ایزوپیپریتنول دهیدروژناز و ایزوپیپریتنول ریداکتاز نشان داد که تنش خشکی باعث ایجاد فرآیند مولکولی شده که منتج به پاسخ‌دهی گیاه با تغییر بیان ژن‌های دخیل در مسیر بیوسنتز ماده مؤثره پولگن می‌گردد.
 

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Expression Analysis of the Key Genes Involved in Biosynthesis Pathway of Major Component of Ziziphora persica Bunge Essential Oil under Drought Stress

نویسندگان [English]

  • Mozhdeh Jalilifar 1
  • Khosro Mehdikhanlou 2
  • Lila Nejadsadeghi 2
  • Qing-Yan Shu 3

1 PhD Student in Production Engineering and Plant Genetics, Assistant Professor, Department of Plant Production and Genetics, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Assistant Professor, Department of Plant Production and Genetics, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Key Laboratory of Plant Resources/Beijing Botanical Garden, Institute of Botany, Chinese Academy of Sciences, Beijing, China

چکیده [English]

Introduction
Iranian Kakoti (Ziziphora persica Bunge) which belongs to the mint genus is widely distributed in Iran. In some parts of Iran, it is used as a spice and condiment in food. Iranian Kakoti is also a medicinal plant with antioxidant and antimicrobial properties. Iranian Kakoti is used in traditional medicine as a stomach disinfectant and cold remedy.  Since the effect of stresses on the increase of secondary metabolites of plants has been proven, the aim of this study was to investigate the effect of drought stress on the secondary metabolites of Iranian Kakoti in comparison with normal conditions by assessment of the expression of genes which are related to the most important secondary metabolites of the studied plant. In previous studies, it was found that pulegone has the largest share among the metabolites produced in this plant.
 
 
 
Materials and Methods
Using the KEGG database, the metabolic pathway and genes involved in the production of the pulegone include the four key genes Isopiperitenol dehydrogenase, Limonene synthase, limonene hydroxylase and Isopiperitenone reductase were identified. Due to the lack of genomic information of Iranian Kakoti, the sequence of the four studied genes was predicted by the sequences of these genes in families close to this plant. The predicted sequences were used for primer design. The plants were cultivated under drought stress and normal conditions. The stress condition was achieved by reaching 30% Field Capacity. After observing phenotypic changes due to stress, the amount of proline and also Leaf relative water content (RWC) measured for the plants in both control and stress condition to confirm the accruing of the stress. Then leaf samples were collected. RNA extraction and cDNA synthesis were performed according to the manufacturer's instructions. The quantity and quality of the obtained RNA were evaluated by Nanodrop and gel electrophoresis. The qualified RNA samples were used for cDNA synthesis according to the manufacturer's protocols. Then the comparing the genes expression under drought stress with the control sample was performed.
 
Results and Discussion
In response to drought stress, leaf RWC was decreased to 31% in the plants under stress condition and proline levels increased by 1.9-fold compared to control value. Analysis of real-time PCR data showed that. The expressions of limonene synthase, limonene hydroxylase, Isopiperitenon dehydrogenase, Isopiperitenon reductase were increased 115, 400, 50 and 120 folds compared to control respectively.
 
Conclusion
The measurement of leaf RWC and proline were found to be beneficial methods to detection of incidence of drought stress in plant.  The expression of genes involved in pulegone synthesis compared to the control plants changes with drought stress. The implement of drought stress is a useful method to increase essential oil in Iranian Kakoti.
 

کلیدواژه‌ها [English]

  • Gene expression
  • Real time-PCR
  • Proline
  • Plant molecular response
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تولیدات گیاهی
دوره 45، شماره 4
بهمن 1401
صفحه 463-474
فایل ها
هم رسانی
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