بررسی بیان ژن‌های کلیدی مسیر بیوسنتز لیمونن تحت تأثیر تنش خشکی در زرین‌گیاه (Dracocephalum kotschyi) در دو مرحله رویشی و گلدهی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 مربی دانشگاه پیام نور، گروه کشاورزی دانش آموخته دکتری دانشگاه بین المللی امام خمینی قزوین

2 دانشیار دانشگاه بین المللی امام خمینی (ره) قزوین

3 استاد، گروه اصلاح نباتات و بیوتکنولوژی، دانشکده کشاورزی، دانشگاه شهرکرد، شهرکرد، ایران

4 قزوین دانشگاه بین المللی امام خمینی (ره)

5 استادیار، گروه بیولوژی، دانشکده علوم، دانشگاه رازی کرمانشاه، کرمانشاه، ایران

چکیده

هدف: زرین‌گیاه با نام علمی Dracocephalum kotschyi یکی از گیاهان دارویی بومی ایران، متعلق به خانواده لامیاسه می‌باشد. از جمله مهم‌ترین مونوترپن‌های شناسایی شده در این گیاه لیمونن است. یکی از عوامل مؤثر بر میزان ترکیبات مونوترپنی در گیاهان دارویی تنش خشکی می‌باشد. هدف از این بررسی تأثیر تنش خشکی در مراحل مختلف رشدی بر بیان ژن‌های کلیدی مسیر بیوسنتز لیمونن در زرین گیاه بود.
مواد و روش‌ها: این آزمایش به صورت فاکتوریل با دو عامل تنش خشکی شامل تنش شدید (25% ظرفیت زراعی)، متوسط (50% ظرفیت زراعی) و کم (75% ظرفیت زراعی) و عدم اعمال تنش (100% ظرفیت زراعی) و مراحل رشدی شامل دوره رویشی و گلدهی در قالب طرح کاملاً تصادفی با سه تکرار انجام شد. آنالیز متابولیت‌های ثانویه و اندازه‌گیری میزان لیمونن با استفاده از دستگاه GC/MS به روش فضای فوقانی و اندازه­گیری بیان نسبی ژن‌های کلیدی مؤثر در بیوسنتز لیمونن با استفاده ازReal Time PCR  صورت گرفت.
نتایج: آنالیز متابولیت‌های ثانویه مقدار لیمونن را در دوره رویشی 57/20 درصد و در دوره گلدهی 41/13 درصد نشان داد. تنش خشکی باعث افزایش معنی‌دار بیان ژن‌های HMGR و DXS در مرحله گلدهی شد، در حالی‌که در مرحله رویشی تغییر معنی‌داری بر بیان این دو ژن‌ مشاهده نشد. در هر دو دوره رشدی تنش خشکی کم باعث افزایش بیان ژن HDR شد، هرچند که در دوره گلدهی این افزایش بیان معنی‌دار نبود، اما تنش خشکی شدید بیان ژن HDR را کاهش داد، هرچند که این کاهش بیان در دوره رویشی معنی‌دار نبود. تنش خشکی باعث کاهش معنی‌دار بیان ژن LS در مرحله گلدهی و افزایش معنی‌دار بیان آن در مرحله رشد رویشی شد.
نتیجه‌گیری: نتایج حاصل از این تحقیق، میزان لیمونن و بیان ژن‌های مؤثر در بیوسنتز آن را تحت تأثیر تنش خشکی وابسته به مرحله رشدی گیاه نشان داد.

کلیدواژه‌ها


عنوان مقاله [English]

The Expression of Limonene Biosynthesis Pathway Key Genes under Drought Stress in Dracocephalum kotschyi at Vegetative and Flowering Stages

نویسندگان [English]

  • Soghra Kiani 1
  • Jafar Ahmadi 2
  • Behroz Shiran 3
  • Sedigheh Fabriki ourang 4
  • Hossein Fallahi 5
1 payame noor university, faculty of agriculture, iran, shahrekord, farokhshar
2 Professor, Department of Genetics and Plant Breeding, Faculty of Agriculture, Imam Khomeini International University, Qazvin, Iran.
3 Professor of Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Shahrekord University, Shahrekord, Iran
4 Associate Professor, Department of Genetics and Plant Breeding, Faculty of Agriculture, Imam Khomeini International University, Qazvin, Iran
5 Assistant Professor, Department of Biology, School of Sciences, Razi University, Bagh-e-Abrisham Kermanshah, Iran
چکیده [English]

Objective
 Dracocephalum kotschyi is an endemic medicinal plant in Iran belonging to the Lamiaceae family. One of the most important monoterpenes identified in this plant is limonene. Drought stress is one of the factors affecting the content of monoterpene in medicinal plants.  In this study, the effect of different levels of drought stress was investigated on the expression of some genes involved in limonene biosynthetic pathway in D. kotschyi at two growth stages.
 
Materials and Methods
 This experiment was conducted as a factorial experiment with two factor included drought stress and growth stages in a completely randomized design with three replications. Drought stress consisted of severe (25% FC), moderate (50% FC), low (75% FC) stress and non-stress or control (100% FC) and growth stages consisted of vegetative and flowering. Secondary metabolites were analyzed and the amount of limonene was measured by GC / MS using Head-space method and the relative expression of key genes involved in limonene biosynthesis was measured using Real Time PCR.
 
Results
 Analysis of secondary metabolites showed, one of the main monoterpenes was limonene whose amounts was 20.57 % and 13.41% during vegetative and flowering stages, respectively. Drought stress significantly increased HMGR and DXS gene expression during flowering stage, while during vegetative stage no significant change was observed on expression of both genes. In both growth stages, low drought stress increased HDR gene expression, although at flowering stage this increase was not significant but severe drought decreased HDR gene expression, although this decrease was not significant at vegetative stage. Drought stress decreased LS gene expression during flowering stage and increased it during vegetative stage.
 
Conclusion
 The results indicated that limonene content and expression of limonene biosynthesis key genes in D. kotschyi under drought stress are growth stage-specific.

کلیدواژه‌ها [English]

  • Dracocephalum kotschyi
  • Drought stress
  • DXS
  • HMGR
  • LS
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