معرفی یک پروتکل کشت بافت برای گیاه لاله واژگون (Fritillaria raddeana) با استفاده از ریزنمونه‌های بذر و سوخ

نوع مقاله: مقاله کامل

نویسندگان

1 دانشجوی دکتری مهندسی علوم باغبانی (گیاهان زینتی)، دانشکده علوم کشاورزی، پردیس دانشگاهی دانشگاه گیلان، رشت

2 استادیار، دانشکده علوم کشاورزی، پردیس دانشگاهی دانشگاه گیلان، رشت

3 استاد، گروه زراعت و اصلاح نباتات پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

4 کارشناس ارشد بیوتکنولوژی، بانک گیاهی، مرکز ذخایر ژنتیکی و زییتی ایران، جهاد دانشگاهی تهران

چکیده

لاله واژگون گرگانی (Fritillaria raddeana Regel.) از گیاهان پیازی خانواده سوسن و یکی از زیباترین گیاهان دنیا است. این گونه بومی ایران، ترکمنستان، به­طور معدود پاکستان و افغانستان می­باشد. دارای ارزش زینتی- دارویی و تحمل نسبی به خشکی و مناطق سنگلاخی و شیب­دار می­باشد. به­منظور معرفی و اهلی­سازی این گیاه بومی، خودرو و در معرض خطر انقراض، نخستین قدم، بررسی روش­های مختلف تکثیر از جمله ریزازدیادی آن است. این پژوهش یک سیستم کارآمد برای باززایی غیرمستقیم از کشت بذر و سوخ، در محیط درون شیشه­ای ارائه می­دهد. اندام­های گیاه پس از برداشت از گیاه یا استخراج سوخ از خاک در طبیعت، ضدعفونی­شده و به‌عنوان ریزنمونه استفاده شدند و تأثیر غلظت­‌های مختلف سایتوکینین (کاینتین، تیدیازورون، بنزیل­آدنین) و اکسین (ایندول­استیک­اسید، ایندول­بوتیریک­اسید، نفتالین­استیک­اسید) بر پینه­زایی و باززایی مورد ارزیابی قرار گرفت. تلفات کمتر، آلودگی بسیار پایین و درصد پینه­زایی بالاتر از مزایای ریزنمونه بذر و وزن پینه بیشتر و درصد باززایی بالاتر در بازه زمانی یکسان، از امتیازات ریزنمونه فلس سوخ تشخیص داده ­شد. همچنین اکثر این ریزنمونه­ها در محیط کشت موراشیگ و اسکوگ دارای 5/0 میلی­گرم در لیتر ایندول­بوتیریک-اسید همراه با 2 میلی­گرم در لیتر تیدیازورون در مسیر باز­زایی قرار گرفتند. 

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Introducing a tissue culture protocol for Fritillaria (Fritillaria raddeana) via seed and bulb scales

نویسندگان [English]

  • Solaleh Salahi Sadr 1
  • Hedayat Zakizadeh 2
  • Mohammad Reza Naghavi 3
  • Jamal-Ali Olfati 2
  • Kian Hazrati 4
1 Ph. D. Student, Department of Horticultural Sciences, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran
2 Assistant Professor, Department of Horticultural Sciences, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran
3 Professor, Department of Agronomy and Plant Breeding, College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
4 M.Sc., Plant GeneBank, Iranian Biological Resource Centre (IBRC), ACECR, Tehran, Iran
چکیده [English]

Fritillaria raddeana is one of the most fascinating flowers, belonging to Lilliaceae family, native to Iran, Turkmenistan, rarely found in Afghanistanand Pakistan. It is an important ornamental-medical bulbous plant, tolerant to arid conditions and stony slopes but facing extinction, so in vitro propagation of this plant will have a great importance for introduction and domestication of this wild and endangered plant. The present study was carried out to find an efficient system for in vitro callus induction and regeneration of F. raddeana via culturing of seeds and bulb scales. These explants were sterilized first and then cultured on media containing various concentrations of plant growth regulators. Different proportions of Auxins (6- indole-3-aceticacid (IAA), α-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA), Dichlorophenoxyacetic acid (2,4-D))  and Cytokinins (Benzylaminopurine (BAP), Thidiazuron (TDZ), Kinetin (KIN)),  and different MS basal mediawere tested. More survival percentage, less contamination and higher percentage of callus formation were the advantages of seed explants and heavier callus with higher regeneration percentage during the same period of time were the positive points of bulb scales explants. The best callus formation and regeneration medium was MS medium supplemented with 2 mgl-1 TDZ + 0.5 mgl-1 IBA.

کلیدواژه‌ها [English]

  • Callus formation
  • endemic plant
  • indirect regeneration
  • plant growth regulators
  1. Aggarwal, D. & Barna, K. S. (2004). Tissue culture propagation of elite plant of Aloe vera Linn. Journal of Plant Biochemistry & Biotechnology, 13(1), 77-79.
  2. Bajaj, Y. P. S., Furmanowa, M. & Olzowska, O. (1988). Biotechnology of the micropropagation of medicinal & aromatic plants. In: Bajaj YPS (Ed), Biotechnology in Agriculture & Forestry 4, Medicinal & Aromatic Plants I (pp.15–103.) Springer, Heidelberg.
  3. Baranova, M. (1981). The ecologo-morphological peculiarities of the underground organs of the representatives of the genus Fritillaria (Liliaceae). Botanicheskii Zhurnal. 66 (10), 1369- 1387.
  4. Baskin, C. C. & Baskin, J. M. (2014). Germination ecology of seeds with morpho-physiological dormancy. Seeds. (2nd ed.) 5 (pp.119-143.) Elsevier Inc.
  5. Bryan, J. E. (2002). Bulbs. Revised edition. Timber Press Inc., Portland, Oregon.
  6. Carasso, V. & Mucciarelli, M. (2014). In vitro bulblet production & plant regeneration from immature embryos of Fritillaria tubiformis Gren. & Godr. Propagation of Ornamental Plants, 14(3), 101-111.
  7. Debiasi, C., Silva, C. G. & Pescador, R. (2007). Micropropagation of Aloe vera L., Revista Brasileria de Plantas Medicinias, 9, 36-43.
  8. Erisen, S., Yorgancilar, M., Atalay, E. & Baboaglu, M. (2010). Prolific shoot regeneration of Astragalus cariensisPlant Cell, Tissue and Organ Culture. 100, 229-233.
  9. Fay, M. F. (1992). Conservation of rare & endangered plants using in vitro methods. In Vitro Cellular & Developmental Biology-Plant, 28, 1-4.
  10. Fridborg, G., Pedersen, M., Landstrom, L. E. & Eriksson, T. (1978). The effect of activated charcoal on tissue cultures, adsorption of metabolites inhibiting morphogenesis. Plant Physiology; 43, 104–6.
  11. Gamborg, O. L., Miller, R. & Ojima, K. (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental cell research, 50(1), 151-158.
  12. Ghahreman, A. & Attar, F. (1999). Biodiversity of plant species in Iran, The vegetation of Iran, plant species, red data of Iran, endemic species, rare species, species threatened by extinction (Vol. 1). Central Herbarium of Tehran University, Faculty of Science.
  13. Gharehyazie, B., Ebrahimie, E., Mohammadi Dehcheshmeh, M., Irannegad, A., Sardari, M. & Salahi Ardacany, A. (2006). Development of commercial protocol for in vitro micropropagation of Fritillaria. International system for agricultural science and technology. Record ID=IR2008000622. Retrieved 2006, Agricultural biodiversity in FAO, fromhttp://www.fao.org/biodiversity.
  14. Gholami, M. (2007). Micropropagation of inverted tulip (Fritillaria imperialis L.). MSc. Thesis, University of Avicenna. Hamedan, Iran. (in Farsi)
  15. Guo, B., Abbasi, B. H., Zeb, A., Xu, L. L. & Wei, Y. H. (2011). Thidiazuron: a multi-dimensional plant growth regulator. African Journal of Biotechnology, 10(45), 8984-9000.
  16. Hamidoghli, S., Chamani, E., Hamidoghli, Y. & Talei, N. (2013). Effect of different plant growth regulators on direct bulblet regeneration from scale explants of Fritillaria imperialis. Journal of Crop Protection, 5(16), 211-217. (in Farsi)
  17. Hao, Y. R., Li, M. S. & Wu, Y. W. (1982). Callus induction & plant regeneration in tissue culture of Fritillaria pallidiflora. Acta Botanica Sinica, 2, 38-43. (in Chinese with English summary)
  18. Helsper, J. P., Bücking, M., Muresan, S., Blaas, J. & Wietsma, W. A. (2006). Identification of the volatile component(s) causing the characteristic foxy odor in various cultivars of Fritillaria imperialis L. (Liliaceae). Plant Research International, Journal of Agricultural and Food Chemistry, 54 (14), 5087-5091.
  19. Jones, M., Cao, J., O’Brien, R., Murch, S. J. & Saxena, P. K. (2007). The mode of action of thidiazuron: auxins, indoleamines, & ionchannels in the regeneration of Echinacea purpurea L. Plant Cell Reports, 26(9), 1481-1490.
  20. Joshi, S. K., Dhar, U. & andola, H. C. (2007) In vitro bulblet regeneration & evaluation of Fritillaria roylei Hook. A high value medicinal herb of the Himalaya, Acta horticulturae, 756, 75-84.
  21. Kamari, G. (1996). Fritillaria species (Liliaceae) with yellow or yellowish-green flowers in Greece. Bocconea 5, 221-238.
  22. Nisyawati, K. K. (2013). Effect of ascorbic acid, activated carbon & light duration on explant browning of banana cultivar barangan (Musa acuminata L.) in vitro culture. International Journal of Recent Research and Applied Studies, 16, 118-123.
  23. Kizil, S., Khawar, K. M. & Sesiz, U. (2013). Bulblets regeneration from in vitro improved leaf of Fritillaria imperialis L. and F. persica L. Current Opinion in Biotechnology, (24), S 39.
  24. Kulkhanova, D. S., Erst, A. A. & Novikova, T. I. (2015). In vitro regeneration from bulbous scales of Fritillaria sonnikovae, an endemic species. Russian Journal of Developmental Biology, 46(4), 215-221.
  25. Le Nard, M. & De Hertogh, A. A. (1993). The physiology of flower bulbs. Elsevier, Amsterdam, (pp. 718- 739). The Netherlands.
  26. Li, S. L., Lin, G., Chan, S. W. & Li, P. (2001). Determination of the major isosteroidal alkaloids in bulbs of Fritillaria by high-performance liquid chromatography coupled with evaporative light scattering detection. Journal of Chromatography A (909), 207-214.
  27. Liao, Z., Chen, M., Tan, F., Sun, X. & Tang, K. (2004). Micropropagation of endangered Chinese aloe. Plant Cell, Tissue & Organ Culture, 76(1), 83-86.
  28. Mirdeilami, Z., Heshmati, Gh. & Mazandarani, M. (2012). Introduction to the flora, life forms & geographic diversity of medical plants of arid & semi-arid regions of North-East of Golestan province. Journal of Iranian Plant Eco-physiological Research, 7(26), 27-37. (in Farsi)
  29. Mohammadi Deh-cheshmeh, M., Khalighi, A. & Naderi, R. (2007). Indirect somatic embryogenesis from petal explant of endangered wild population of Fritillaria imperialisPakistan Journal of Biological Sciences, 10(11), 1875-1879.
  30. Mohammadi Deh-cheshmeh, M., Khalighi, A., Naderi, R., Sardari, M. & Ebrahimie, E. (2008). Petal: a reliable explant for direct bulblet regeneration of endangered wild populations of Fritillaria imperialis L. Acta Physiologiae Plantarum, 30(3), 395-399.
  31. Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth & bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473-497.
  32. Murch, S. J. & Saxena, P. K. (2002). Mammalian neurohormones: potential significance in reproductive physiology of St. John's wort (Hypericum perforatum L.) The Science of Nature - Naturwissenschaften, 89(12), 555-560.
  33. Murthy, B. N. S., Murch, S. J. & Saxena, P. K. (1998). Thidiazuron: A potent regulator of in vitro plant morphogenesis. In Vitro Cellular & Developmental Biology-Plant, 34(4), 267-275.
  34. Özcan, S., Parmaksız, I., Mirici, S., Çöçü, S., Uranbey, S., İpek, A., Sancak, C., Sarihan, E.O., Gürbüz, B., Sevimay, C. S. & Arslan, N. (2007). Efficient in vitro bulblet regeneration from immature embryos of endemic & endangered geophyte species. In: Z. Xu. (Ed), Biotechnology & Sustainable Agriculture 2006 & Beyond. (pp. 381-383). Springer Science.
  35. Palmer, C. D. & Keller, W. A. (2011). Plant regeneration from petal explants of Hypericum perforatum L. Plant Cell, Tissue & Organ Culture, 105 (1), 129-134.
  36. Petrić, M., Jevremović, S., Trifunović, M., Tadić, V., Milošević, S., Dragićević, M. & Subotić, A. (2013). The effect of low temperature and GA3 treatments on dormancy breaking and activity of antioxidant enzymes in Fritillaria meleagris bulblets cultured in vitro. Acta Physiologiae Plantarum, 35(11), 3223-3236.
  37. Pretto, F. R. & Santarem, E. R. (2000). Callus formation & plant regeneration from Hypericum perforatum leaves. Plant Cell, Tissue & Organ Culture, 62 (2), 107-113.
  38. Rechinger, K. H. (1990). Fritillaria L. In: K.H.  Rechinger (Ed), Flora Iranica., 165 (pp. 61–76.) Akademische druck -u-verlagsantalt. Graz.
  39. Rix, E. M. (1977).  Fritillaria L. (Liliaceae) in Iran. Iranian Journal of Botany, 1(2), 75-95.
  40. Subotic, A., Trifunović, M., Jevremović, S.  & Petrić, M. (2010). Morpho-histological study of direct somatic embryogenesis in endangered species Frittilaria meleagrisBiologia Plantarum, 54(3), 592-596.
  41. Sudha, C. G. & Seeni, S. (1994). In vitro multiplication & field establishment of Adhatoda beddomei C. B. Clarke, a rare medicinal plant. Plant Cell Reports, 13, 203-207.
  42. Thomas, T. D. (2008). The role of activated charcoal in plant tissue culture. Biotechnology Advances, 26(6), 618-631.
  43. Witomska, M. & Lukaszewska, A. (1997). Bulbet regeneration in vitro from different explants of Fritillaria imperialis. Acta Horticulturae, 430, 331-338.
  44. YanLi, C., Yuan, C., FengXia, G., YuHong, L. & Tian, L. (2010). A study on soaking & germination characteristics of Fritillaria przewalskii seeds. Acta Prataculturae Sinica, 19(4), 41-46.
  45. Yaojia, Z., Hongmei, W. & Ruilian, Z. (1994). A preliminary study on the ecological characteristics & germinating law of the seeds of Fritillaria yuzhongensis in the Maxian mountain region of Gansu Province [China]. Acta Ecologica Sinica (China), Agricultural biodiversity in FAO. Retrieved 1992from http://www.fao.org/ biodiversity.
  46. Yu, J., Wei, J. H., Chen, S. L., Dai, Y. & Yang, C. M. (2008). Dormancy & germination characteristics of Fritillaria cirrhosa seed [J]. Chinese Traditional & Herbal Drugs7, 1081-1084.
  47. Zhiliang, L. & Feng, P. Y. L. (1989). Identification of inhibitor in the aqueous extract of Fritillaria polliflora Schrek seeds [J]. China Journal of Chinese Materia Medica5, 004.
  48. Zhou, B., Wei, X., Wang, R. & Jia, J. (2010). Quantification of the enzymatic browning & secondary metabolites in the callus culture system of Nigella glandulifera Freyn et Sint. Asian journal of Traditional Medicine, 5(3), 109-116.
  49. Ziv, M. & Lilien-Kipnis, H. (2000). Bud regeneration from inflorescence explants for rapid propagation of geophytes in vitro. Plant Cell Reports, 19 (9), 845-850.