نوع مقاله : مقالات پژوهشی

نویسندگان

دانشگاه محقق اردبیلی

چکیده

سوسن چلچراغ (L. ledebourii) کمیاب‌ترین گونه‌ی جنس سوسن است که بیشتر در ناحیه‌ی قفقاز می‌روید. ایران نیز یکی از مهمترین مناطق پراکنش این گونه در حال انقراض است. لزوم توجه به حفظ این گیاه و استفاده از آن در کارهای اصلاحی و انتقال ژن، نیاز به باززایی فراوان آن در شرایط درون شیشه‌ای دارد. بدین منظور در این تحقیق سعی شد تا با استفاده از تنظیم کننده‌های رشد گیاهی و یک تنش غیرزیستی (فراصوت) توانایی پیازچه‌زایی و ریشه‌زایی این گیاه مورد بررسی قرار گیرد. این پژوهش به صورت آزمایش فاکتوریل در قالب طرح کاملاً تصادفی در چهار تکرار صورت گرفت. نتایج حاصل از پژوهش نشان داد که تنظیم‌کننده‌ی رشد NAA نقش مهمی در افزایش وزن کل پیازچه‌های تولید شده دارد، به‌طوری‌که بیشترین مقادیر این صفت از غلظت 1/0 و 1 میلی گرم در لیتر NAA حاصل شد. همچنین بیشترین تعداد ریشه در هر پیازچه از ترکیب تیماری 01/0 میلی گرم در لیتر BA بدست آمد. در ترکیب غلظت‌های مختلف هورمون NAA و هورمون BA، بیشترین وزن کل پیازچه‌ها در ریزنمونه‌ی پیازچه در محیط کشت MS حاوی 1/0 میلی گرم در لیتر NAA بدون BA بدست آمد. همچنین در غلظت بالای BA نسبت به NAA، همه‌ی سطوح فراصوت باعث افزایش تعداد فلس‌ها و وزن پیازچه‌ها گردید و بیشترین تعداد پیازچه‌ها از 10، 20 و 30 ثانیه فراصوت‌دهی حاصل شد. اثر مثبت تیمار فراصوت بر پیازچه‌زایی و صفات مربوط به آن نشان می‌دهد که این تیمار می‌تواند تاثیر زیادی برای باززایی این گیاه در شرایط درون شیشه‌ای داشته باشد. بنابراین از آنجا که فراصوت به طور غیرمستقیم بر میزان هورمون‌های درون‌زا در ریزنمونه تاثیر می‌گذارد، توصیه می‌گردد که در کنار تیمار هورمونی می‌توان از تیمار فراصوت در تکثیر این گیاه با ارزش استفاده کرد و با استفاده از آن میزان باززایی از این گیاه را افزایش داد.

کلیدواژه‌ها

عنوان مقاله [English]

Effect of Plant Growth Regulators and Ultrasound on the Bulblet Production and Root Induction in Lilium ledebourii

نویسندگان [English]

  • Mehdi Mohebodini
  • Zahra Azimzadeh
  • Esmaeil Chamani
  • Malihe Erfani

university of mohaghegh ardabili

چکیده [English]

Introduction: Lily (L. ledebourii) is the rarestspeciesof thegenusLilium, and grows in Caucasus region. Iranis one of the important distribution areas of this endangered species. It is important as an ornamental plant due to its large and attractive white flowers that are equal to those of commercial lilies in terms of beauty.The two main constraints on growing this plant are a low multiplication rate and the high cost of bulb production. Five to ten flowers commonly appear on each plant, even specimens with up to 15 flowers have been observed. Plant tissue culture techniques are widely used in plant propagation and using these methods can effectively provide micro-propagation of this plant in large scale. High percentage ofregeneration is necessary for plant protection, using in the breeding programs and gene transfer to this plant. Therefore, the effect of plant growth regulators and abiotic stress (ultrasound) werestudied on the bulblet production and root induction of Lilium ledebourii.
Materials and Methods: The experiment was factorial based on completely randomized design with four replicattions and was carried out in tissue culture lab of University of MohagheghArdabili in 2015. For this purpose, segmentsof scale explant was treated with ultrasound and cultured on MS medium supplemented with different concentrations of NAA and BA alone and/or in combination with each other. In this experiment, different concentrations of NAA (0, 0.01, 0.1 and 1 mgl-1) and BA (0, 0.01, 0.1 and 1 mgl-1) and different Ultrasound exposure duration (0, 5, 10, 20 and 30 second) were studied. In order to remove possible contamination from the media, all media were autoclaved for 20 minutes at 121 °C. At the end of the experiment, the number of bulblet, root length, fresh weight of bulblet were recorded. The cultures were kept at 25±2°C under illumination with daylight fluorescent lamps (30 mol m-2s-1) at 16 h photoperiod. Data was subjected for analysis of variance and compare means using SPSS 16.

Results and Discussion: The results showed that ultrasound had negative effect onroot length, so that the highest root length was observed in explants without ultrasound treatment. Result also indicated that ultrasound had positive effect on bulblet production and root induction. A different effect of growth regulators was observed in similar media on the bulblet formation percentage. The 0.1 NAA concentration had a higher efficiency while increasing NAA insignificantly decreased bulblet induction. The highest total weight and number of bulblets obtained by 0.1 mgl-1 NAA. Concentrations of NAA increased rooting percentage. Different concentrations of NAA had also significant effects on some traits. So that, the highest weight of bulblets obtained by 0.01 and 0.1 mgl-1 BA and the highest number of roots obtained in control. Bulblet maximum mean weightwas in30 seconds ofultrasoundtreatment, which hada significantdifference with the control treatment (without ultrasoundtreatment). In the other hand, ultrasound increased the number and weight of bulblets.Mechanical stress and microstreaming by acoustic cavitation might be considered as the most possible cause of the various physiological effects of ultrasound on cells. The enhancement of V-ATPase transport and ATP hydrolysis activities seem to be an ultrasound-induced metabolic response of cells. High-intensity ultrasound is well known to be destructive to biological materials, disrupting the cell membranes and deactivating biological molecules such as enzymes and DNA. Low-intensity ultrasound, on the other hand, has shown a range of sub lethal biological effects that are of potential significance in biotechnology. There are several processes that take place in the presence of cells or enzymes activated by ultrasonic waves. High-energy ultrasonic waves break the cells and denature the enzymes. Low-energy ultrasound can modify cellular metabolisms or facilitate the uptake of nutrient, and make them easily through the cellular walls and membranes. In the case of enzymes, the increase in the mass transfer rate of the reagents to the active site seems to be a most important factor.
Conclusions: The results showedthatthebulblet production at first stages and a little root formation in tissue culture is useful for fast bulblet inductionandthenrooting. Finally, it seems that ultrasound in combination with plant growth regulators have the potential to produce the highest average number of bulblets in the scale explant.

کلیدواژه‌ها [English]

  • Benzyladenine
  • In vitro
  • Naphthalene acetic acid
  • Rooting
1- Atrashi M., Tavakoli E., Darzi M.T., Hashemi J., Rozbeh S.H., and Masum M. 2011. Effect of ultrasound on the production of Carvone as a secondary metabolite in callus derived from Bunium persicum Boiss. Journal of Herbal Drugs, 2: 129-135. (in Persian)
2- Azadi P., and Khosh-Khui M. 2007. Micropropagation of Lilium ledebourii (Baker) Boiss as affected by plant growth regulator, sucrose concentration, harvesting season and cold treatments. Electronic Journal of Biotechnology, 4: 582-591.
3- Bakhshaie M., Babalar M., and Khalighi M. 2010. Somatic embryogenesis and plant regenerationof Lilium ledebourii (Baker) Boiss., an endangered species. Plant Cell Tissue and Organ Culture, 102: 229–235.
4- Bochu W., Jiping S., Biao L., Jie L., and Chuanren D. 2004. Soundwave stimulation triggers the content change of the endogenous hormone of the Chrysanthemum mature callus. Colloids and Surfaces B: Biointerfaces, 37: 107-112.
5- Bochu W., Yoshikoshi A., and Sakanishi A. 1998. Carrot cell growth response in a stimulated ultrasonic environment. Colloids and Surfaces B: Biointerfaces, 12: 89-95.
6- Gaba V., Kathiravan K., Amutha S., Singer S., Xiaodi X., and Ananthakrishnan G. 2006. The uses of ultrasound in plant tissue culture. Journal Plant Tissue Culture Engineering, 417-426.
7- Ibanescu M., Constantinovici D., and Strajeru S. 2008. Study about the ultrasound effects on potato plantlets (Solanum tuberosum L.), conserved in vitro through slow growth method. Studia Universitatis, 18: 41-44.
8- Joersbo M., and Brunstedt J. 1990. Direct gene transfer to plant protoplasts by mild sonication. Plant Cell Reports, 9: 207-210.
9- Kanchanapoom K., Ponpiboon T., Wirakiat W., and Kanchanapoom K. 2011. Regeneration of lily (Lilium longiflorum ‘Easter lily’) by callus derived from leaf explants cultured in-vitro. Science Asia, 37: 373–376.
10- Kapoor R., Kumar S., and Kanwar J.K. 2008. Bulblet regeneration from ex-vitro root explant in lily hybrids. Horticultural Science, 35: 107-112.
11- Kapoor R., Kumar S., and Kanwar J.K. 2009. Bulblet production from node explant grown in-vitro in hybrid lilies..International Journal of Plant Production, 3: 1-6.
12- Liu Y., Takatsuki H., Yoshikoshi A., Wang B., Sakanishi A. 2003. Effects of ultrasound on the growth and vacuolar H+-ATPase activity of Aloe arborescens callus cells. Colloids and Surfaces B: Biointerfaces, 32: 105-116.
13- Mir J.I., Ahmed N., Itoo H., Sheikh M.A., Rashid R., and Wani S.H. 2012. In-vitro propagation of Lilium (Lilium longiflorum). Indian Journal of Agricultural Sciences, 82(5): 455–558.
14- Novak, F.J., and Petru, E. 1981. Tissue culture of Lilium hybrids. Scientia Horticulturae, 14: 191-199.
15- Omidi M, and Farzin N. 2012 Biotechnology approaches for improvement of medicinal plants. Modern Genetics Journal 7:209-220 (in Farsi).
16- Padasht Dahkaei, M. N., Khalighi, A., Naderi, R., and Mousavi, A. 2006. Effect of temperature, propagation media and scale position on bulblet regeneration of Chelcheragh lily (Lilium ledebourii) by scaling method. Seed and Plant 22: 383-397. (In Persian)
17- Padasht Dahkaei M. N., Khalighi A., Naderi R., and MoUsavi A. 2008. Effect of different concentration of benzyladenine (BA) and naphthalene acetic acid (NAA) on regeneration of Lilium ledebourii (Chelcheragh Lily) using bulblets microscales. Seed and Plant Improvement Journal, 24: 321-332. (in Persian)
18- Pelkonen V.P., and Kauppi A. 1999. The effect of light and auxins on the regeneration of lily (Lilium regale Wil.) cells by somatic embryogenesis and organogenesis. International Journal of Plant Sciences, 160: 483-490.
19- Pierik R.L., and Steegmans H.M. 1975. Effect of auxins, cytokinins, gibberellins, abscisic acid and etephon on regeneration and growth of bulblets on excised bulb scale segments of hyacinth. Physiology Plant, 34: 14-17.
20- Podwyszynska M. 2006. Improvement of bulb formation in micropropagted tulips by treatment with NAA and pacleobutrazol or accymidol. Acta Horiculture, 725: 679-684.
21- Shin Y.K., Abdullahil Baque M., Elghamedi S., Lee E.J., and Paek K.Y. 2011. Effects of activated charcoal, plant growth regulators and ultrasonic pretreatments on in-vitro germination and protocorm formation of Calanthe hybrids. Australian Journal of Crop Science, 5: 582-588.
22- Tatari varnusfaderani M., Fotouhi Ghazvini R., Hamidoghli Y., and Hatamzadeh A. 2004. Effects of plant growth regulators and kind of bulblet initiation on in-vitro culture of bulb scale segments from Lilium ledebourii (Chelcheragh lily). Journal of Agricultural Science, 1(2): 19-27. (in Persian)
23- Una G., Sanin H., Jasmina C., Anisa R., and Damir M. 2010. In-vitro propagation of Lilium martagon L. var. cattaniae Vis. and evaluation of genotoxic potential of its leaves and bulbs extracts. Acta Biologica Slovenica, 53(2): 53- 60.
24- Wei M., Yang C.Y., andWei S.H. 2012. Enhancement of the differentiation of protocorm like bodies of Dendrobium officinale to shoots by ultrasound treatment. Journal of Plant Physiology, 169: 770-774.
25- Xiujuan W., Bochu W., Yi J., Danqun H., andChuanren D. 2003. Effect of sound stimulation on cell cycle of chrysanthemum (Gerbera jamesonii). Colloids and Surfaces B: Biointerfaces, 29: 103-107.
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