بهینه‌سازی تولید کالوس و باززایی درگیاه زینتی زامیفولیا (Zamioculcas zamiifolia)

نوع مقاله : مقالات پژوهشی

نویسندگان

دانشگاه آزاد اسلامی واحد لاهیجان

چکیده

امروزه پرورش و تکثیر گیاهان زینتی به خصوص گیاهانی که تکثیر آنها به سختی صورت می­گیرد، از طریق کشت بافت از ارزش و اهمیت زیادی برخوردار است. تکثیر گیاه زینتی زامیفولیا با ارزش اقتصادی بالا از طریق قلمه برگ و تقسیم ریزوم به سختی صورت می‌گیرد و به مدت زمانی حدود 7 ماه نیاز دارد لذا جهت تسریع در تکثیر آن کشت بافت مفید واقع می‌گردد. جهت بهینه­سازی تولید کالوس و باززایی، ریزنمونه‌های دمبرگ، ریزوم، برگ و شاخه، در محیط کشت­های با سطوح متفاوت تنظیم کننده‌های رشد در دو آزمایش جداگانه به صورت فاکتوریل در قالب طرح کاملاً تصادفی در سه تکرار مورد بررسی قرار گرفتند. در آزمایش اول ریزنمونه‌های دمبرگ و ریزوم در سه مرحله کالوس، شاخه و ریشه‌زایی به ترتیب در سطوح هورمونی ] BA (4، 2، 0 میلی‌گرم بر لیتر)، 2,4-D (2، 1، 0 میلی‌گرم بر لیتر)[، ] BA (2، 1، 0 میلی‌گرم بر لیتر) و 2,4-D (1، 5/0، 0 میلی‌گرم بر لیتر)[ و ] BA (2، 1، 0 میلی‌گرم بر لیتر) و NAA (1، 5/0، 0 میلی‌گرم بر لیتر)[ قرار گرفتند و در آزمایش دوم ریزنمونه‌های برگ و شاخه نیز در سه مرحله کالوس، شاخه و ریشه‌زایی به ترتیب در سطوح هورمونی ] BA (4، 2، 0 میلی‌گرم بر لیتر) و NAA (1، 5/0، 0 میلی‌گرم بر لیتر)[ ، BA] (2، 1، 0 میلی‌گرم بر لیتر) و NAA (1، 5/0، 0 میلی‌گرم بر لیتر)[ و] BA (2، 1، 0 میلی‌گرم بر لیتر) و NAA (2، 1، 0 میلی‌گرم بر لیتر)[ قرار گرفتند. در مرحله کالوس‌زایی، صفات مدت زمان تا شروع کالوس‌زایی و پس از گذشت 5 هفته درصد کالوس‌زایی و وزن ترکالوس اندازه­گیری گردید. در مرحله باززایی مدت زمان تا شروع ریشه‌زایی، مدت زمان تا شروع شاخه‌زایی و پس از گذشت 5 هفته طول شاخه و تعداد برگ اندازه‌گیری گردید. بالاترین درصد کالوس‌زایی، کوتاهترین زمان برای رسیدن به کالوس و بیشترین وزن تر کالوس در محیط کشت با هورمون­های 2 میلی‌گرم بر لیتر BA و 1 میلی‌گرم بر لیتر NAA در ریز نمونه برگ به ترتیب با 5/94 درصددرصد، 14 روز و 1/1 گرم از آزمایش دوم مشاهده گردید. بهترین تیمار در مرحله شاخه‌زایی با کوتاهترین زمان تا شروع شاخه‌زایی، بلندترین طول شاخه و بیشترین تعداد برگ به ترتیب با 5/10 روز ، 10/4 سانتی متر و 8 برگ مربوط به ریزنمونه برگ با غلظت هورمونی 2 میلی‌گرم بر لیتر BA و 5/0 میلی‌گرم بر لیتر NAA از آزمایش دوم بود و در مرحله ریشه‌زایی بهترین تیمار برای ریزنمونه دمبرگ با غلظت هورمونی 1 میلی‌گرم بر لیتر BA و 5/0 میلی‌گرم بر لیتر NAA با 14 روز زمان تا ریشه‌زایی از آزمایش اول بود. از نتایج آزمایش‌ها در مرحله باززایی چنین بر آمد که نوع ریزنمونه در شاخه‌زایی و ریشه‌زایی همانند مرحله کالوس‌زایی موثر بوده‌است به طوری که در مرحله شاخه‌زایی نتایج به دست آمده از ریزنمونه ریزوم نسبت به ریزنمونه دمبرگ بهتر بود اما در مرحله ریشه‌زایی ریزنمونه دمبرگ بهتر از ریزنمونه ریزوم عمل کرده ‌است.

کلیدواژه‌ها


عنوان مقاله [English]

Optimization of Callus Production and Regeneration of Zamiifolia (Zamioculcas zamiifolia)

نویسندگان [English]

  • M. Sayadi Nejad
  • S.M. Sadeghi
Lahijan Branch, Islamic Azad University, Lahijan, Iran
چکیده [English]

Introduction: Zamiifolia is a perennial ornamental plant and is one of the most important medicinal plants of the Aracea family. The origin of this evergreen, low-anticipated plant is East Africa. Zamiifolia spreads through the leaves and split rhizomes, which is very time-consuming. The traditional Zamiifolia proliferation method have been done by dividing rhizomes and leaf cuttings, but the production efficiency is low due to the slow growth of the plant, tubers and roots.  In addition, due to the warm and humid environment, reproduction is limited to summer season. Due to the traditional reproductive problems in this plant, tissue culture or microbial culture is the best way to replicate rapidly and to achieve a large number of plants with the same genetic structure, as well as the elimination of diseases in the short term and reducing the costs. The aim of this study was to compare different microorganisms in terms of calogenesis and regeneration, as well as to determine the optimum culture medium for Zamiifolia tissue culture.
Materials and Methods: In this study, the explants prepared for the first experiment, including rhizome and petiole and the explants for the second experiment were the leaeaves and shoots. In the first experiment, rhizome and petiole were cultured in three replications in ½ MS medium containing BA (0, 2, 4 mg / L) and 2,4-D (0,1,2 mg / L) in combination with vitamins, 30 g/l sucrose, 5 g / L agar and adjusted to 5.8 PH. The cultivars were cultured for the callus induction under temperature of 27-25 ° C and light conditions of 16 hours light and 8 hours darkness. After 5 weeks, the percentage of callus and fresh callus weight were measured. The callus generated from rhizome and petiole in three replicates on ½ MS medium containing BA (0, 1, 2 mg / L) and 2,4-D (0, 0.5, 1 mg / l) for shoots and after the observation of branches and leaf buds were grafted on to ½ MS medium containing BA (0, 1, 2 mg / L) and NAA (0, 0.5, 1 mg / L) for rooting. Traits such as time to shoot elongation were recorded at regeneration stage, and after 5 weeks, shoot length and the number of leaves were measured. The time to rootstock was also recorded. In the second experiment leaf and shoot explants were cultured in ½ MS medium containing BA (0, 2, 4 mg / l) and NAA (0, 0.5, 1 mg / L) in combination with vitamins, 30 g/l sucrose, 5 g / L agar and PH adjusted to 5.8. The cultivars were cultured for the callus induction under temperature 27-25 °C and light conditions of 16 hours light and 8 hours darkness. The time to reach the callus was recorded and after 5 weeks, the percentage of callus and fresh callus weight were measured. The calli generated from the leaves and shoots were cultures on ½ MS medium containing BA (0, 1, 2 m g / L) and NAA (0, 0.5, 1 mg / L) for shoots and after observation of branch and leaves were transplanted to the ½ MS medium containing BA (0, 1, 2 mg / L) and NAA (0, 1, 2 mg /L) for rooting. The traits such as time to shoot elongation were recorded at the regeneration stage, After 5 weeks, the shoot length and the number of leaves were measured.The time to rootstock was also recorded.
Results and Discussion: The results of the first experiment showed that the effect of the rhizome and petiole type on the callus formation was significant at 1% level. So that the rhizome showed greater ability to callogenesis. The results of the second experiment showed that the effect of the type leaf and shoot on the callus formation was significant at 1% level. So that the leaf showed greater ability to callogenesis. The highest percentage of callosing (94.5%), the shortest time to reach the callus (14 days) and the highest callus weight (1.1 g) in culture medium with 2 mg / l BA and 1 mg / l hormones NAA was observed in leaf samples from the second experiment. The best treatment in the shoot elongation stage, which included the shortest time to shoot formation (10.5 days), the longest shoot length (4.10 cm), and the highest leaf number (8 leaves) in the leaf extract with hormonal concentrations of 2 mg / 0 mg / L NAA was observed from the second experiment. In the rooting stage, the best treatment for petiole extracts with hormonal concentrations was 1 mg / l BA and 0.5 mg / l NAA with 14 days to rooting from the first experiment.
Conclusion: In this study, explants and various concentrations of growth regulators had significant effect on the response to callus induction in Zamiifolia. In the first experiment, the rhizome and in the second experiment the leaf showed a better reaction to callus induction. According to this research, it can be suggested that the treatments applied in both experiments should be applied on all four leaves, petiole, rhizome and shoot samples, and the best culture type and the best culture medium for the cultivation of Zamiifolia plant tissue should be determined in subsequent studies.

کلیدواژه‌ها [English]

  • Tissue culture
  • Micro propagation
  • Sub culture
  • BA
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