ریزازدیادی لاله واژگون (.Fritillaria imperialis L) از طریق روش باززایی غیرمستقیم در شرایط کشت درون شیشه‌ای

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی ارشد، گروه علوم باغبانی، دانشگاه کشاورزی و منابع طبیعی رامین خوزستان

2 استاد، گروه باغبانی، دانشگاه کشاورزی و منابع طبیعی رامین خوزستان

چکیده

ایران یکی از خاستگاه‌های لاله واژگون (Fritillaria imperialis L.) است. این گیاه زینتی در معـرض انقراض می‌باشد. بنابراین افزایش آن از طریق کشت بافت ضروری است. این پژوهش در چهار آزمایش: تولید پینه، سوخک‌زایی، ریشه‌زایی و سازگاری انجام گردید. در آزمایش اول از دو ریزنمونه فلس و برگ اولیه در محیط کشت MS با غلظت های مختلف NAA و IBA اقدام به تولید پینه گردید. نتایج نشان داد بیشترین وزن پینه از ریزنمونه فلس در محیط کشت MS حاوی 0/5 میلی‌گرم در لیتر NAA و 0/5 میلی‌گرم در لیتر IBA تولید گردید. در آزمایش دوم از پینه های به‌دست آمده، در محیط کشت MS با غلظت‌های مختلف TDZ و KIN، پس از 6 و 12 هفته، تعداد سوخک مورد بررسی قرار گرفت. نتایج نشان داد، بیشترین تعداد سوخک بعد از 12 هفته از پینه حاصل از ریزنمونه فلس در محیط کشت MS  حاوی 0/5 میلی‌گرم در لیتر NAA و سپس در محیط کشت اندام زایی با 0/5 میلی‌گرم در لیتر TDZ همراه با 0/5 میلی‌گرم در لیتر KIN تشکیل شد. سپس سوخک‌ها به محیط کشت MS با غلظت‌های مختلف NAA (صفر، 0/5، 1 و 2 میلی‌گرم در لیتر) جهت ریشه‌زایی منتقل شدند. نتایج نشان داد، بیشترین تعداد ریشه از سوخک‌های حاصل از ریزنمونه برگ اولیه، در محیط کشت حاوی 1 میلی‌گرم در لیترNAA  به‌دست آمد. در‌نهایت سوخک‌های ریشه‌دار شده پس از شستشو با آب مقطر جهت سازگاری به گلدان‌های حاوی پرلیت،کوکوپیت، پرلیت و کوکوپیت که در اتوکلاو گندزدایی شده بودند، منتقل شدند. نتایج نشان داد سوخک‌های حاصل از ریزنمونه فلس در بستر کشت کوکوپیت، دارای بیشترین تعداد ریشه بود. این تحقیق نشان داد که استفاده از کشت بافت، نقش مهمی در جلوگیری از انقراض لاله واژگون دارد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Utilization of Indirect Regeneration to Induce Micropropagation of Fritillaria imperialis L. on in vitro Culture

نویسندگان [English]

  • Mojtaba Rahimi 1
  • Mohammad Hosein Daneshvar 2
چکیده [English]

Background and Objectives
Iran is home of one of the most delightful and graceful flower of Fritillaria in the world. Its habitation is commonly expanded on the vast areas: growing in beds, borders, backgrounds and rock gardens. Besides, it is also suitable for naturalizing and excellent for cut flowers. Fritillaria imperialis as a medicinal plant has pharmaceutical purpose in which possesses a health beneficial essence that used in production of mucus and cough medicine. In addition, extractions of plant tissues contain certain of properties which affect to reduce blood pressure and blood sugar. Unfortunately, the habitation of this ornamental plant is under threat and majority of its habitation already being to ruin due to human activities and urbanization. The propagation of Fritillaria through offsets (bulblets) is prolonging approach which takes approximately 3-4 years. Whereas, using tissue culture technique for flower production is required less time probably within a few months and economically has a huge advantage due to low cost and a numerous plant production in a short time.
Materials and methods
This research was carried out in four experiments: callus formation, bulblet production, root formation and acclimatization. In the present study, a factorial experiment was assigned as completed randomized design (CRD) with three replications. In the first experiment, leaf primordia and scale explants were cultured on MS medium supplemented with different concentrations of NAA and IBA in order to produce callus formation. In the second experiment, the calli were cultured on organogenesis medium which were contained MS medium supplement, with kin. and TDZ, after 6 and 12 weeks. In the third experiment, the bulblets were transferred into rooting media, which were contained MS medium supplemented with different concentrations of NAA (0, 0.5, 1.0 and 2.0 mg/l). In the fourth experiment, after washing the bulblet whit warm distilled water (40ᵒC) that contain 2.0 mg/l benlate fungicide, they then were planted into the pots containing sterilized perlite, cocopeat and perlite along with cocopeat.
Results
In the first experiment, the results demonstrated that the highest weight of callus (3.29g) was sustained in scale explants on basal MS medium which was supplemented with 0.5mg/l NAA along with 0.5mg/l IBA. In second experiment, the result indicated that the numerous of bulblets were developed on MS medium supplemented with 0.5mg/l TDZ along with 0.5mg/l kin. Which of these, the callus scale were produced on MS medium supplemented with 0.5mg/l NAA. In the third experiment, in order to retain rooting the bulblets, they were cultured on MS medium supplemented with different concentration of NAA. The result elucidated that the large quantity of roots (7.7) were obtained from leaf primordia on MS medium supplemented with 1.0mg/l NAA. In the fourth experiment, rooted bulblets were washed with warm distilled water for 5-10min, and then they were transferred into pots with perlite, cocopeat and perlite + cocopeat.
Discussion
The results revealed information that the rooted bulblets from scale growing on cocopeat substrate had retained the most diameter (21.64mm) as compared to other treatments. The finding results under this investigation strongly suggested that using micropropagation it is definitely a possible trend to abolish the barriers which caused to deteriorate of Fritillaria imperiallis.
 

کلیدواژه‌ها [English]

  • Callus
  • Fritillaria imperialis L
  • Bulblet regeneration
  • micropropagation
  • In vitro culture
References
  1. Chen, U.C., Tai, C.D., Chen, C.C., and Tsay, H.S. 2000. Studies on the tissue culture of Fritillaria hupehensis. Influence of medium component and light treatment on in vitro rooting and acclimatization on bulblet. Journal of Agricultural ResearchChina, 49(4): 48-55.
  2. Daneshvar, M.H. 1992. Callus induction and organogenesis in cultivar of peach (Prunus persica L.). Ph.D. Thesis, Department of Agriculture and Horticulture Faculty of Agricultural and Food Sciences. University of Nottingham Sutton Bonington, UK, 329 P.
  3. Danut, P.K. and Bhojwani, S. S. 1995. In vitro corm formation and field evaluation of corm- derived plants of gladiolus. Scientia Horticulture, 61: 115-129.
  4. De Hertogh, A. and Lenard, M. 1993. The physiology of flower bulbs. Elsevier, pp. 718-739.
  5. Kim, Y.J., Hasegawa, P.M., and Bressan, R.A. 1981. In vitro propagation of Hyacinth. Hort Science, 16(5): 645-647.
  6. Nhut, D.K. 1998. Micropropagation of lily (Lilium longiflorum) via in vitro stem node and pseudo-bulblet culture. Plant Cell Reports, 17: 913-916.
  7. Nhut, D.T., Le, B., Tanaka, M., and Van, T.T. 2001. Shoot inducation and plant regeneration from receptacle tissue of Lilium longiflorom. Scientia Horticulture, 87: 131-138.
  8. Ozel, A. and Erden, K. 2005. Determination of some morphological properties of Fritillaria imperialis L. and F. persica L. Fourth Congress of Agriculture in Southeast Anatolia Project Areas, 1562-1567.
  9. Rahimi, M., Daneshvar, M.H., Heidari, M., and Yari, F. 2013. In vitro micropropagation of Fritillaria imperialis L. through induction of indirect organogenesis. International Journal of Agronomy and Plant Production, 4(3): 418-424.
  10. Salehzadeh, S.H., Daneshvar, M.H., and Moallemi, N. 2008. Indirect organogenesis from scale, leaf primordia and immature floret explant of hyacinth (Hyacinthus orientalis L.). American-Eurasian Journal of Agricultural & Environmental Sciences, 4(5): 640-645.
  11. Squires, W.M., Langton, F.A., and Fenlon, J.S. 1991. Factors influencing the transplantation success of micropropagated Narcissusbulbil. Journal of Horticultural Science, 66: 661-671.
  12. Wang, S.H., Gaoa, W., Chena, H., and Xiao, P. 2005. New starches from Fritillaria species medicinal plants. Carbohydrate Polymer, 61: 111-114.
  13. Witomska, M. and Lukaszewska, A.J. 1997. Bulblet regeneration in vitro from different explants of Fertillaria imperialis. Acta Horticulturae, 430: 331-338.

5.        Hussy, G. 1975. Totipotency in tissue explants and callus of some members of Liliaceae, Iridaceae and Amarylidaceae. Journal of Experimental Botany, 26: 645-647.

7.        Mohammadi-Dehcheshmeh, M., Khalighi, A., Naderi, R., Sardari, M., and Ebrahimie, E. 2008. Petal: A reliable explant for direct bulb let regeneration of endangered wild populations of Fritillaria imperialis L. Acta Physiologiae Plantarum, 30: 395-399.

12.    Rahman, A.U., Akhtar, M.N., Choudhary, M.I., Tsuda, Y., Sener, B., Khalid, A., and Parviz, M. 2002. New steroidal alkaloids from Fritillaria imperialis and their cholinesterase inhibiting activities. Chemical and Pharmaceutical Bulletin, 50: 1013-1016.

14.    Santos, A., Fidalgo, F., Santos, I., and Salema, R. 2002. In vitrobulb formation of Narcissus asturiensis, a threatened species of the Amaryllidaceae. International Journal of Horticultural Science and Technology, 77: 149-152.

15.    Shiau, Y.J., Tai, C.D., Chen, C.C., and Tsay, H.S. 2000. Studies on the tissue culture of Fritillaria hupehensis Hosiaoet K.C. Hsia. I. The induction of bulb scale and embryogenic callus from different source explant. Journal of Agricultural Research of China, 49(4): 29-38.

16.    Simmonds, J.A. and Cumming, B.G. 1976. Propagation of Lilium hybrids. Production of plantlets from bulb-scale callus culture for increased propagation rate. Scientia Horticulture, 5: 161-170.

20.    Yang, S.R. Tai, C.D., Chen, C.C., and Tsay, H.S. 2001. Effect of plant growth regulators and status of the medium on induction and proliferation of protocorm-like-bodies and bulblet from bulb scale culture of Fritillaria hupehensis. Hsiao ET. K. C. Hsia, Journal of the Agricultural Association of China, 5: 414-422.