Callus induction and plant regeneration of Juniperus excelsa using in vitro technique

Document Type : Research article

Author

Research institute of Forests and Rangelands, Tehran, Iran

Abstract

Low germination and non-viability of embryos are the main hindrans for forestation of Juniperus excelsa. Conventional methods of asexual plant propagation of this species have not been successful so far. In order to in vitro propagation, small pieces of stem of 8-10 years old trees(10-15 mm long) containing shoot tips with needles fascicles were used as explants. Those were excised from the plants and after surface sterilization. Samples were cultured on a Murashige & Skoog (MS) medium and six revised MS medium supplemented with different concentrations of BAP, Kin, IBA, NAA and 2,4-D. Then the cultures were incubated in a climate chamber at temperatures of 25°C (for day) and 15°C (for night) and 12 hours light at 2000-2500 Lux with 75% humidity. Results indicated that callugenesis and callus growth restricted by both type and nitrate and hormonal source. The best simulation of callus growth occurred on cultures of without KNO3, with gloutamin (100 ml/l) and amount of 2,4-D/BAP at more than 1 ratio. Although adventitious buds were formed on juvenile stages of parent plants, but root induction did not occur. In addition, the season of sampling can also restrict the calogenesis. Such, calougenesis of collected plants in autumn was more than in spring. We could not find any shoot proliferation on medium MS, however by elimination of ammonium nitrate from medium MS bud appeared from callus as well as leaf blade.

Keywords

References

- صالحی شانجانی، پ.، 1375.
2- صدری، ح.ع.، نراقی، ط.، 1374. مروری بر ریزازدیادی سوزنی‌برگان با تأکید بر تحقیقات تکثیر گونه ارس به‌ویژه کشت جوانه انتهایی. مجله پژوهش و سازندگی، 29: 50-55.
3- Al-Talib, K.H. and Torrey, J.G., 1959. The aseptic culture of isolated buds of Pesudutsuga taxifolla. Plant Physiol., 34: 630-637.
4- Bonga, J.M. 1977. Organogenesis in in vitro cultures of embryonic shoots.
5- Erikson (14bj) of Abies blasama. In vitro, 13: 41-48.
6- Cheah, K.J. and Cheng, T.Y., 1978. Histological analusis of adventitious bud formation in cultured doglas fir cotyledons. Am. J. Bot., 65:72-108.
7- Gomez. M.P. and Segura, J., 1995. Axillary shoot proliferation in cultures of explantas from mature Juniperus oxycedruws trees. Tree physiology. 16(8): 681-686.
8- Gomez. M.P. and Segura, J., 1996. Morphogenesis in leaf and single-cell cultures of mature Juniperus oxycedruws. Tree physiology. 15(9): 625-628.
9- Ilahi, I., 1986. Biotechnology in agriculture and forestry. Vol. 1. In: Tremblay, F.M., Perinet, P., Lalonde, M., Sakai, A., and Bajaj, Y.P.S. (Eds) Trees I. Springer Verlag Publisher, Berlin.: 321-498.
10- Javeed, Q.N.; Perveen, R.; Imtiazul, H.; Ilahi, I., 1980 Propagation of Juniperus polycarpos C. Koch through tissue culture. I. Induction of callus. Pakistan journal of forestry,30(2): 72-77.
11- Konar, R.N. and Oberoi, Y.P. 1965. In vitro development of embryoids on the cotyledons of Bota orientalls. Phytomorphology, 15: 137-140.
12- Moreira-da-Silva, M.H. and Debegh, P., 1996. Clonal and topophysis effect in axillary shoot development on Juniperus brevifolia (Seub.) Antoine. Mededelingen Faculteit landbouwkundige en Toegepaste Biologische Wetenschappen, Universiteit Gent., 61(1): 145-149.
13- Mahmood, A., Khalida, A. and Khan, N.H., 1992. Bud multiplication in Juniper. Hamdard Medicus,35(4): 51-56.
14- Martel, S., Houde, M., Laliberte, S. and Tremblay, M.F., 2001. Studied the effects of nitrogen source on morphogenetic pattern and glutamine synthetase expression in jack pine meristematic nodules and calli in vitro. Symposium on tree Biotechnology in the Next Millenium, USA.
15- Murashige, T. and Skoog, F., 1962. A resived medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol. 14: 10-16.
16- Negussie, A., 1997. In vitro induction of multiple buds in tissue culture of juniperus excelsa. Forest Ecology and Management, 82(2): 115-123.
17- Welinder, K.G., 1994. Plant peroxidase: Structure-function relationship. Plant peroxidase, 1980-1990.
Iranian Journal of Forest and Poplar Research
Volume 11, Issue 2
June 2003
Pages 247-264

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