ریزازدیادی و روش کپسوله‌کردن-آب‌گیری برای حفاظت فراسرد گیاه لاله‌ی واژگون (Fritillaria imperialis)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه باغبانی، دانشکده کشاورزی، دانشگاه آزاد اسلامی، رشت

2 دانشجوی دکتری علوم باغبانی، گروه باغبانی، واحد رشت، دانشگاه آزاد اسلامی، رشت، ایران

3 دانشیار، گروه باغبانی، واحد رشت، دانشگاه آزاد اسلامی، رشت، ایران

10.22103/jab.2021.16279.1251

چکیده

هدف: لاله­‌‌ی واژگون (Fritillaria imperialis) از خانواده­‌ی سوسنیان (Liliaceae)، یک گونه‌ی زینتی در معرض خطر انقراض است. تکثیر این گیاه در شرایط طبیعی، کارایی بالایی ندارد و زمان­بر است. ریزازدیادی، یک فن مناسب برای تکثیر لاله­‌ی واژگون در شرایط درون‌شیشه‌ای است. برای حفظ خزانه‌ی ژنتیکی این گیاه تهدیدشده، استفاده از فن حفاظت انجمادی ضروری است. برای حفاظت انجمادی، حضور پیش‌تیمارهای مناسب ضروری است. مهم‌ترین و پر استفاده‌ترین پیش‌تیمار، کپسوله‌کردن-آب‌گیری است. هدف از پژوهش حاضر، تکثیر درون‌شیشه‌ای لاله­‌ی واژگون با استفاده از تنظیم­‌کننده­های رشد گیاهی و حفاظت انجمادی آن با استفاده از پیش‌تیمار کپسوله­‌کردن-آب‌گیری بود.
مواد و روش‌ها: فلس‌های سوخ به‌عنوان ریزنمونه و محیط کشت MS به­‌عنوان محیط کشت پایه استفاده شدند. برای ریزازدیادی، کینتین (Kin) و نفتالین استیک اسید (NAA) در غلظت‌های صفر (به­‌عنوان شاهد)، 5/0، 1 و 2 میلی‌گرم در لیتر مورد استفاده قرار گرفتند. برای حفاظت انجمادی، ریزنمونه­‌ها بعد از پیش‌تیمار با روش کپسوله‌کردن-آب­‌گیری، در ازت مایع نگهداری شدند. کپسوله­‌کردن با استفاده از آلژینات سدیم انجام شد. آب‌گیری در هوا و با استفاده از غلظت بالای سوکروز انجام گرفت.
نتایج: نتایج ریزازدیادی نشان داد که بیشترین تعداد برگ (5/3) و تعداد ریشه (7/5) در ریزنمونه­های تیمارشده با 5/0 میلی‌گرم در لیتر Kin همراه با یک میلی­‌گرم در لیتر NAA به‌دست آمد. نتایج حفاظت انجمادی نشان داد که کپسوله‌کردن-آب‌گیری در هوا به‌عنوان یک پیش‌تیمار، نقش مؤثری در بقا و قدرت جوانه‌زنی ریزنمونه‌ها (5/70 درصد) بعد از نگهداری در ازت مایع داشت. کمترین بقای ریزنمونه مربوط به ریزنمونه­‌های شاهد (بدون پیش‌تیمار) بود. گیاهچه‌های باززایی‌شده در شرایط درون‌شیشه­‌ای، درون گلدان‌های حاوی پیت موس و پرلایت (به نسبت 1:1) کاشته شدند و با شرایط محیطی سازگار گردیدند. نرخ زنده‌مانی در گیاهچه‌های ریزازدیادی‌شده، 90 درصد و الگوی رشد گیاهان باززایی‌شده شبیه گیاهان مادری بود.
نتیجه‌گیری: پژوهش حاضر، استفاده از 5/0 میلی‌گرم در لیتر Kin همراه با یک میلی‌گرم در لیتر NAA را برای ریزازدیادی و کپسوله‌کردن-آب‌گیری را برای حفاظت انجمادی ژرم­‌پلاسم لاله‌ی واژگون پیشنهاد می‌کند. ریزازدیادی توسط اندام‌زایی و جنین‌زایی مستقیم و غیر مستقیم یک رویکرد مناسب برای تکثیر گونه‌های زینتی در حال انقراض است. موفقیت این روش‌ها به نوع و غلظت اکسین‌ها و سیتوکینین‌های به­‌کار برده شده در محیط کشت، انفرادی یا در ترکیب با یکدیگر، بستگی دارد. به‌دلیل حفاظت از گیاهان زینتی در معرض خطر انقراض، نیاز است که روش‌های زیست­‌فناوری نوین به کار برده شود.

کلیدواژه‌ها


عنوان مقاله [English]

Micropropagation and encapsulation-dehydration method for cryopreservation of Fritillaria imperialis

نویسندگان [English]

  • Shima Seydi 1
  • shahram sedaghathoor 2
  • behzad kavyani 3
1 Department of Horticulture, Faculty of Agriculture, Islamic Azad University, Rasht
2 Department of Horticultural Science, Rasht Branch, Islamic Azad University, Rasht, Iran.
3 Associate Professor, Department of Horticultural Science, Rasht Branch, Islamic Azad University, Rasht, Iran.
چکیده [English]

Objective
Fritillaria imperialis from the Liliaceae family is an ornamental species in danger of extinction. Therefore, it is nessessary to conserve the plant genetic pool. Two proper methods to access this goal are micropropagation and germplasm conservation in in vitro freezing conditions. The presence of suitable pre-treatments is necessary for cryopreservation. The most important and the most application of pre-treatment is encapsulation-dehydration. The purpose of current research was in vitro proliferation of F. imperialis using plant growth regulators and its cryopreservation using encapsulation-dehydration pre-treatment. 
Materials and methods
Bulb scales as explants and MS medium as basic culture medium were used. For micropropagation, kinetin (Kin) and α_naphthaleneacetic acid in concentrations of 0 (as control), 0.5, 1 and 2 mg l–1 were applied. For cryopreservation, explants were pretreated with encapsulation-dehydration followed by conservation in liquid nitrogen. Encapsulation was done using sodium alginate. Dehydration was carried out in air and using high level of sucrose.
Results
The results of micropropagation showed that the maximum number of leaf (3.5) and root number (5.7) was obtained in explants treated with 0.5 mg l–1 Kin along with 1 mg l–1 NAA. The results of cryopreservation showed that encapsulation-dehydration in air as a pre-treatment had effective role on the survival and regeneration of explants (70.5%) after conservation in liquid nitrogen. Least explant survival was obtained in control (without pre-treatment). In vitro regenerated plantlets were cultivated in plastic pots containing peat moss and perlite (in ration of 1:1) and acclimatized with environmental conditions. The survival rate was 90% and the growth pattern of regenerated plants was similar to that of mother plants.
Conclusions
Present research proposes the use of 0.5 mg l–1 Kin together with 1 mg l–1 NAA for micropropagation and encapsulation-dehydration for germplam cryopreservation of F. imperialis. Micropropagation by direct and indirect organogenesis and embryogenesis is a proper approach for proliferation of ornamentals in danger of extiction. The success of these methods depends on type and concentrations of auxins and cytokinins applied in culture medium, singular or in combination. In order to protect rare and endangered ornamental species, modern biotechnological tools need to be utilized.

کلیدواژه‌ها [English]

  • growth regulators
  • Liliaceae family
  • Endengered Plants
  • Germplasm conservation
  • In vitro culture
 
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