اثر تیمارهای مختلف تنظیم‌کننده‌های رشد، نوع ریزنمونه و روشنایی بر تولید کالوس گیاه داروئی گزنه (Urtica dioica L.)

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانشیار، گروه تولید و ژنتیک گیاهی (بیوتکنولوژی)، دانشکده کشاورزی، دانشگاه جهرم، جهرم، ایران

2 دانشجوی سابق کارشناسی، گروه بیوتکنولوژی، دانشکده کشاورزی، دانشگاه جهرم، جهرم، ایران

3 استادیار، گروه تولید و ژنتیک گیاهی (بیوتکنولوژی)، دانشکده کشاورزی، دانشگاه جهرم، جهرم، ایران

چکیده

چکیده
گیاه گزنهبه‌عنوانیکی از گیاهان با ارزش داروئی بوده که به علت دارا بودن ترکیبات فیتوشیمیایی با ارزش، اخیراً توجه زیادی به آن برای درمان بیماری‌هایمختلف و همچنین استفاده از آن در صنایع غذایی به‌عنوان نگهدارنده شده است. در این پژوهش به منظور بهینه‌سازی شرایط کشت بافت برای القاء کالوس از بافت‌های مختلف این گیاه، اثر تیمارهای مختلف تنظیم‌کنندههایرشد، شرایط نگهداری محیط کشت و نوع ریزنمونه بر میزان تولید کالوس آن مورد بررسی قرار گرفت. به این منظور آزمایشی به‌صورت فاکتوریل در قالب طرح کاملاً تصادفی با سه تکرار در سال های 1395-1396 در دانشگاه جهرم انجام گردید و ریزنمونه‌های مختلف شامل برگ‌هایمسن و جوان، گره و بذر این گیاه در محیط کشت MS جامد حاوی 30 گرم بر لیتر ساکارز و غلظت‌های متفاوت دو تنظیم‌کننده رشد تو-فور-دی (1، 2 و 3 میلی‌گرم در لیتر) و بنزیل آدنین (1، 5/0 و 1/0 میلی‌گرم در لیتر) کشت شدند و چهار هفته پس از کشت صفاتی از قبیل درصد تولید کالوس، زمان کالوس‌دهی، بافت و رنگ کالوس‌های القایی مورد ارزیابی قرار گرفتند. بر اساس نتایج حاصله، ریزنمونه‌های مورد استفاده تفاوت معنی‌داری از نظر تولید کالوس نشان دادند، به‌طوری‌که ریزنمونه‌های برگ مسن، برگ جوان و گره قادر به تولید کالوس بودند. پاسخ ریزنمونه‌ها برای تولید کالوس با قرار گرفتن در شرایط نوری مختلف متفاوت بود به نحوی که شرایط تاریکی تأثیربیشتری در القای کالوس و میزان رشد آن داشت. همچنین اثر تیمارهای مختلف تنظیم‌کننده رشد بر تولید کالوس ریزنمونه‌های مختلف در سطح یک درصد معنی‌دار بود، به‌طوری‌که بیشترین میزان تولید کالوس در ریزنمونه‌های برگ‌های جوان و ریزنمونه‌های گره در محیط‌ کشت دارای یک میلی‌گرم در لیتر تو-فور-دی و یک میلی‌گرم بنزیل آدنین به‌دست‌آمد ولی در مورد برگ مسن بیشترین میزان این صفت در یک میلی‌گرم در لیتر تو-فور-دی و 1/0 میلی‌گرم در لیتر بنزیل آدنین در شرایط تاریکی حاصل شد. در بین ریزنمونه‌های مورد استفاده، ریزنمونه‌های برگ جوان در شرایط تاریکی بیشترین درصد تولید کالوس را نشان داده و رشد کالوس بهتری داشتند. استفاده از ریزنمونه برگ جوان و تیمار یک میلی‌گرم در لیتر تو-فور-دی و یک میلی‌گرم در لیتر بنزیل آدنین و نگهداری کشت‌ها در شرایط تاریکی برای تولید کالوس از این گیاه پیشنهاد می‌گردد.
 

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Effect of Different Plant Growth Regulators, Explant Types and Light on Callogenesis Traits of Medicinal Plant Nettle (Urtica dioica L.)

نویسندگان [English]

  • Abdolkarim Zarei 1
  • Fatemeh Salajeghe 2
  • Mojahed Kamalizadeh 3
1 Associate Professor, Department of Plant Production and Genetic (Biotechnology), Faculty of Agriculture, Jahrom University, Jahrom, Iran
2 M.Sc. Graduate of Biotechnology, Faculty of Agriculture, Jahrom University, Jahrom, Iran
3 Assistant Professor, Department of Plant Production and Genetic (Biotechnology), Faculty of Agriculture, Jahrom University, Jahrom, Iran
چکیده [English]

Abstract
 
Background and Objectives
Nettle (Urtica dioica L.) is considered a valuable plant with high level of various beneficial phytochemical, which has been used centuries in traditional medicine. Since it contains various secondary metabolites and shows noticeable activity against different Gram-positive and Gram-negative bacteria, nettle has been identified as an appropriate choice for wider applications in the food and pharmaceutical industries in the recent years. This plant is native to Asia, Europe, Africa and North America and grows naturally in different regions of Iran. Although nettle is an important medicinal plant worldwide, there is no information about optimization of its tissue culture conditions. This study was carried out to investigate the effects of different plant growth regulators, explant types and light condition on the callogenesis properties of nettle plant under in vitro condition.
 
Materials and Methods
A factorial experiment was carried out on the base of completely randomized design with three replicates. Different explants including young leaves, old leaves, nodes and seeds of nettle plant were cultured in the solid MS medium supplemented with 30 g l-1 sucrose and different concentrations of plant growth regulators of 2,4-D (1, 2 and 3 mg l-1) and BA (0.1, 0.5 and 1 mg l-1). Callus-related characteristics, including time of callus induction, callogenesis percentage, and their color and texture were analyzed. Analysis of variance was carried out by SAS software ver. 9.3.1 and mean comparison was conducted using Duncan multiple range test.
 
Results
According to the results, callus induction was significantly influenced by explant type. The highest callus induction was attained from young leaves, nodes and old leaves, respectively. The results also showed that callus induction and its properties were significantly influenced by light condition. Preservation of cultures in dark condition resulted in higher callus induction than light condition. In addition, the effect of plant growth regulators was significant on callus induction and its properties. The highest percentage of callus induction was obtained from young leaf explants in the medium containing 1 mg l-1 2,4-D and 1 mg l-1 BA in dark condition followed by node explants in the same plant growth regulator treatment. However, medium supplemented with 1 mg l-1 2,4-D and 0.1 mg l-1 BA showed to be more effective for old leaf explants. Based on the results of this research, young leaf explants in the MS medium containing 1 mg l-1 2,4-D and 1 mg l-1 BA in dark condition are suggested for callus induction from nettle plant.
 
Discussion
Type, color and amount of produced callus in different plant species are highly dependent on the genotypes, compositions of tissue culture media as well as the levels of both endogenous and exogenous plant growth regulators. Auxin and cytokinin are two main plant growth regulators playing important roles in callogenesis of different plant species. There is an interaction between the level of plant growth regulators and color of produced callus. Based on the results of present study, it seems that in the nettle plant, node explants endure a higher level of plant growth regulators and hence produce a brighter callus than the leaf segment explants when they are treated with high concentrations of these chemicals.
 

کلیدواژه‌ها [English]

  • 6-Benzyladenine
  • Callogenesis
  • Culture media
  • Factorial experiment
  • 2
  • 4-D
 
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