تأثیر هورمون‌های رشد بر باززایی درون شیشه‌ای زنبق مردابی (Iris pseudacorus)

نوع مقاله : مقالات پژوهشی

نویسندگان

دانشگاه محقق اردبیلی

چکیده

به‌منظور انجام این طرح، سه آزمایش در قالب طرح کاملاً تصادفی در آزمایشگاه کشت بافت گروه باغبانی دانشگاه محقق اردبیلی انجام شد. برای باززایی گیاه از بذر، از غلظت‏‏های مختلف NaOH(5، 10، 15و20 مولار) و خراش‏دهی با کاغذ سمباده (خراش‏دهی نرم، خراش‏دهی متوسط و خراش‏دهی سخت) استفاده شد. نتایج حاصل از مقایسه میانگین داده‏‏ها نشان داد که غلظت 20 مولار NaOH و نیز خراش‏دهی سخت با کاغذ سمباده بیشترین میزان جوانه‏زنی را داشتند. دو ماه پس از جوانه‌زنی بذور، هیپوکوتیل حاصل از دانهال‌های بذری طی دو آزمایش جداگانه تیمار شدند که در آزمایش اول غلظت‌های 1، 2 و 4 میلی‏گرم بر لیتر از هورمون‏های اکسین (پیکلرام و 2,4-D) و در آزمایش دوم غلظت‌های 1، 2 و 4 میلی‏گرم بر لیتر سایتوکینین (TDZ و BA) با 4 تکرار برای هر تیمار، در قالب طرح کاملاً تصادفی کشت شدند. نتایج حاصل از تجزیه واریانس داده‏ها نشان داد که بین تیمار‌ها در مقایسه با شاهد تفاوت معنی‏داری در سطح احتمال 1 درصد وجود داشت. نتایج حاصل از مقایسه میانگین داده‏ها در پارامترهای رشد کالوس نشان داد که غلظت 4 میلی‏گرم بر لیتر پیکلرام و 1 میلی‏گرم بر لیتر بیشترین تأثیر را بر القای کالوس در این گیاه داشت. نتایج حاصل از مقایسه میانگین داده‏ها در پارامترهای رشد گیاهچه نشان داد که تیمار BA با غلظت1 میلی‏گرم بر لیتر بیشترین میزان شاخه‌زایی را در این گیاه تولید نمود.

کلیدواژه‌ها


عنوان مقاله [English]

Investigation on the Hormone Effects on in vitro culture of Iris pseudacorus

نویسندگان [English]

  • E. Chamani
  • M. Taheri
Mohaghegh Ardabili University
چکیده [English]

Three experiments were conducted in tissue culture and biotechnology laboratory of Horticulture Department of Mohaghegh Ardabili University in 2012. For the regeneration of plant from seed, different concentrations of NaOH (5, 10, 15, 20 M) and various scarification methods with sandpaper (soft scarification, medium scarification and hard scarification) were used based on completely randomized design with 4 replications. The results of experiment revealed that seeds treated by 20 M NaOH and hard scarification produced the highest germination rate. After 2 months of seed germination, hypocotyles of seeds were used as explants and cultured in MS medium containing different concentration of 2,4-D, picloram, TDZ and BA (1, 2, 4 mg/l) based on completely randomized design with 4 replications. Mean comparison revealed that explants treated by 4 mg/l picloram and 1 mg/l 2,4-D produced the highest callus content. Mean comparison showed that explant treated by 1 mg/l BA produced the highest shoots. However, to investigate the soluble protein changes during growth stages and to study the effects of 2,4-D, picloram, TDZ and BA on soluble protein and experiment was conducted. The result showed that by increasing the plant age, soluble protein was reduced and also the highest soluble protein was found after 4 weeks of germination. The result also showed that explants treated by 4 mg/l picloram and 1 mg/l BA produced the highest soluble protein content.

کلیدواژه‌ها [English]

  • Micropropagation
  • Scarification
  • Tissue culture
پیری خ. و نظریان فیروزآبادی ف. 1388. راهنمای کشت بافت گیاهان. انتشارات بوعلی سینا.
2- Baskin J.M., and Baskin C.C. 2003. New approaches to the study of the evolution of physical and physiological dormancy, the two most common classes of seed dormancy on the earth. The Biology of Seeds,4: 371-380.
3-Beyl C.A., Sharma G.C. 1983. Picloram induced somatic embryogenesis in Gasteria and Haworthia. Plant Cell, Tissue and Organ Culture, 2: 123-132.
4- Blake J., Eeuwens C.J. 1982. Culture of coconut palm tissues with a view to vegetative Propagation. Springer, 9: 145-148.
5- Boltenkov E.V., Rybin V.G. and Zarembo E.V. 2003. Specific Features of cultivation of Iris ensata Thunb. Callus Tissue. Applide biochemistry and microbiology, 40(2): 206-212.
6- Boltenkov E., Zarembo E. 2005. In vitro regeneration and callogenesis in tissue culture of floral organs of the genus Iris (Iridaceae). Biology Bulletin, 32 (2): 138-142.
7- Boltenkov E. V., Mironova L. N. and Zarembo E. V. 2006. Effect of Phytohormones on Plant Regeneration in callus culture of Iris ensata Thunb. Plant Physiology, 34: 446-450.
8- Chen W.H., Davey M.R., Power J.B. and Cocking E.C. 1988. Control and maintenance of plant-regeneration in sugarcane callus cultures. Journal of Experimental Botany, 39:251-261.
9- Chengalrayan K., Abuzid A. and Gallo-Meagher M. 2005. In vitro regeneration of plants from sugarcane seed-derived callus. In vitro Cellular & Developmental Biology, 41:477-482.
10- Ciomas J., Reflini K., Iswandar E. and Forster P. 2011. Effects of Picloram in Inflorescence culture of Oil Palm. Plant Cell Tiss Organ Culture, 10: 71-78.
11- Daffalla H., Abdellatef E., Elhadi A. and Khalafalla M. 2010. Effect of Growth Regulators on In Vitro Morphogenic Response of Boscia senegalensis (Pers.) Lam. Poir. Using Mature Zygotic Embryos Explants. Biotechnology Research International, 1-8.
12- Demiralay A., Yalcin-Mendi Y., Aka-kacar Y. and Cetiner S. 1998. In vitro propagation of Ficus Carica L. var. Bursa siyahi through meristem culture. Acta Horticulture, 480: 165-167.
13- Fitch M. M., Moore P. H. 1990. Comparison of 2,4-D and picloram for selection of long-term totipotent green callus cultures of sugarcane. Plant Cell Tiss. Organ Cult. 20:157–163.
14- Han J.G., Ni X.Q., Mao P.S., Pu X.C., and Du G.P. 1996. Method to break dormancy in Zoysia japonica seed. Acta Agrestia Sinca, 4: 246-250.
15- Ho W. J., Vasil I. K. 1983. Somatic embryogenesis in sugarcane (Sacharrum officinarum L.). I. The morphology and ontogeny of somatic embryos. Protoplasma, 118:169-180.
16- Huang X.Q., Wei Z.M, 2004. High-frequency plant regeneration through callus initiation from mature embryos of maize (Zea Mays L.). Plant Cell Report, 22:793-800.
17- Johri M.M., Mitra D. 2001. Action of plant hormones. Current Science, 80(2); 199-205.
18- Ichihashi S., Kato S. 1986. Clonal Propagation of Iris kaempferi by means of flower organ culture. Bulltein Aichi University Edu, 35: 135-143.
19- Jarvis B.C., Yasmin S. 1987. Plant growth regulators and adventitious root development in relation to auxin. Biologia Plantarum, 29: 189-198.
20-Jehan H., Courois D., Ehret C., Lerch K. and Petiard V. 1994. Plant regeneration of Iris pallida Lam. and Iris germanica L. via somatic embryogenesis from leaves, apices and young flowers. Plant Cell Reports, 13: 671-675.
21- Jevremovic S., Radojevic L. 2002. Plant regeneration from suspension cultures of Iris pumila L. Acta Horticulture, 572: 59-65.
22- Kamo K. Chen J. and Lawson R. 1990. The establishment of cell suspension cultures of gladiolus that regenerate plants, in vitro cell development. Biol. Plant. 26: 425- 430.
23- Kedra M., Bach A. 2004. Morphogenesis of Lilium martagon L. explants in callus culture. Acta Biologica, 47: 65-73.
24- Keshtkar A.R., Keshtkar H.R., Razavi S.M., and Dalfardi S. 2008. Methods to break seed dormancy of Astragalus cyclophyllon. African Journal of Biotechnology, 7(21): 3874-3877.
25- Kim S.K., Lee J.S., Haung K.H. and Ahn B.J. 2003. Utilization of embryogenic cell cultures for the mass production of bulblets in oriental and easter lilies. Acta Horticulture, 625: 253-259.
26- Kim T., Ahn C., Bae K. and Choi V. 2009. The embryogenic competency and morphological changes during somatic embryogenesis in Iris pseudacorus. Korean Society for Plant Biotechnology and Springer, 3: 251-257.
27- Kumar S., Kashyap M. and Sharma D. R. 2005. In vitro regeneration and bulblet growth from lily bulbscale explants as affected by retardants, sucrose and irradiance. Biologia Plantarum. 49(4): 629 - 632.
28- Kumar O., Rupavati T. 2002. In vitro inductiom of callusogenesis in chilli peper (Capsicum annum L.). International Journal Of Current Reseaech, 3: 42-45.
29- Malik K.A., Saxena P.K. 1992. Thidiazuron induces high frequency of shoot regeneration in intact seedlings of pea (Pisum sativum), chickpea (Cicer arietinum) and lentil (Lens culinaris Medik). Australian Journal of Plant Physiology, 19: 731-740.
30- Mares D.J. 2005. Quarterly reports on plant growth regulation and activities of the PGRSA. 32nd Annual conference the Plant Growth Regulation Society of America, 33(2): 78-89.
31- Mielke K.A. and Anderson W.C. 1989. In vitro bulblet formation in Dutch Iris. Hort science, 24: 1028-1031.
32- Mujib A., Pal A.K. 1995. Inter-varietal variation in response to in vitro cloning of carnation. Crop Research, 10: 190-194.
33- Radojevic L., Subotic A. 1992. Plant regeneration of Iris setosa Pall. through somatic embryogenesis and organogenesis. J. Plant Physiology, 139: 690-696.
34- Ramey V. 2001. Iris pseudacorus. Center for aquaticand Invasive Plants, University of Florida.
35- Rzepka-Plevnes D., Kurek J. 2001. The influence of media composition on the proliferation and morphology of Ficus benjamina plantlets. Acta Horticulture, 560: 473-476.
36- Şener O., Can E., Arslan M. and Çeliktaş N. 2008. Effects of genotype and picloram concentrations on callus induction and plant regeneration from immature inflorescence of spring barley cultivars (Hoerdeum vulgare L.). Biotechnology & Biotechnological Equipment, 22: 915-920.
37- Shibli R.A., Ajlouni M.M. 2000. Somatic embryogenesis in the endemic black iris. Plant Cell Tissue and Organ Culture, 61: 15-21.
38- Shimizu S., Tanaka M. and Mori H. 2009. Auxin-cytokinin interactions in the control of shoot branching. Plant Molecular Biology, 69(4): 429-435.
39- Stoltz L.P. 1968. Iris seed dormancy. Physiol. Plant. 21: 1328-1331.
40- Sun Y., Zhang Y. and Wang K. 2006. NaOH scarification and stratification improve germination of Iris lacteal var. chinensis seed. Hort Science, 41(3): 773-774.
41- Tribulato A., Remotti P., Loeffler H. and Vantuyl J. 1997. Somatic embryogenesis and plant regeneration in Lilium longiflorum Thunb. Plant Cell Reports, 17: 113-118.
42- Villasen˜or CJ., De Lucas MA., Gomez GR. and Mena SJ. 2007. A comparative study of five horizontal subsurface flow constructed wetlands using different plant species for domestic wastewater treatment. Environ Technol. 28: 1333–1343.
43- Wu B.H., Yan J., Zhou Y.H., and Zuo W.X. 1998. Inhibitory effects of seed coat on seed germination in Iris confuse and its hybrid. J. Scichan. Agricultural Science in Finland, 16(3): 337-340.
44- Zare S., Tavili A.and Darini M. 2010. Effects of different treatments on seed germination and breaking seed dormancy of Prosopis koelziana and Prosopis Juliflora. Journal of Forestry Research, 22(1): 35-38.
45-Zheng S., Henken B., Sofiari E., Keizer P., Jacobsen E., Kik C. and Krens F. 1999. Effect of cytokinins and Lines on Plant Regeneration from Long-Term callus and Suspension cultures of Allium cepa, L. Euphytica, 108: 83-90.