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Báo cáo khoa học: "Micropropagation of Araucaria columnaris Hook."

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Micropropagation of Araucaria columnaris Hook. P.K. Khosla L. Sehgal* O.P. Sehgal University of Horticulture and Forestry, Solan 173 230, India DYSP regulators and/or other chemicals. The cultures Introduction kept at 25°C under cool white fluorescent were light (1.5 klux), from 40 W tubes, for 16 h each day. At least 20 replications were used for each The genus Araucaria comprises several treatment. forest tree species, magnificent evergreen including Araucaria columnaris Hook., which has a high ornamental value as well. Conventional propagation by seeds Results is slow and inadequate for producing large uniform progenies. Cuttings are difficult to root and they produce plagiotropic plants The explants, initially cultured on MS except when taken from apical shoots. medium supplemented with kinetin (5-12 Successful micropropagation of this spe- pM) and naphthalene acetic acid (1-3 cies has not been reported. The aim of N were est:ablished on the medium M), this study was to use explants from nor- containing only kinetin (10 pM). The esta- mally growing mature trees for in vitro blishment of cultures was maximum propagation of A. columnaris. (40-50%) when the explants were col- lected in April and November, whereas it was low (10-30%) during other months of the year. The development of axillary buds Materials and Methods obtained by subculturing on a was medium containing 6-benzylaminopurine (BAP, 12.5 pM), indole butyric acid (IBA, 1 of and lateral Tips (3-5 mm) secondary tertiary branches, taken from 14 yr old trees, steri- !M) and 3% sucrose. were lized in mercuric chloride (0.05%) for 1 min and Explants taken for in January, prechilled cultured on Murashige and Skoog’s (MS) at 5°C and cultured on MS medium 1 mo medium (Murashige and Skoog, 1962), contain- ing different concentrations of various growth (containing 10 uM kinetin and 1 !M IBA) * Delhi University, N. Delhi 110049, India. Delhi Permanent address: Gargi College,
produced almost twice the shoot growth Subculturing or direct planting on a medium containing kinetin (10 pM) and rate as compared to the ones not chilled, giving a shoot length of 1.5 cm in 6 wk thiourea (1.3 ,uM) enhanced the shoot length and production of axillary branches (Table I).
latter were separated and Araucaria is hard-to-root material in (Fig. 1The a used for multiplication. Addition of 0.3% success in rooting has culture, and limited activated charcoal to the medium also been reported only for one species viz., indicated a faster rate of growth (Table I). A. cunningharnii (Haines and de Fossard, 1977; Burrows, 1983). Nevertheless, the of shoot growth and axillary production branches obtained in culture in the pres- Discussion and Conclusion ent study is an important step in micro- propagation. Attempts to induce in vitro rooting are in progress. Micropropagation of most of the conifers has not been successful. Out of 6 or so reports on in vitro culture of about 12 spe- cies of Araucaria (Haines and de Fossard, References 1977; Maene & Debergh, 1987), only Bur- rows (1983) used coppice shoots from mature trees. Others have utilized ortho- Bonga J.M. (1981) Organogenesis in vitro of tis- tropic shoots from juvenile plants. Recent- from mature conifers. In vitro 17, 511-518 8 sues ly, Handro (1986) reported the develop- Boulay M. (1979) Multiplication et clonage ra- ment of axillary shoot in vitro, starting from pide du Sequoia sempervirens par la culture in branches of a 20 yr old tree of A. angusti- vitro. In: Etudes et Recherches, AFOCEL, Domaine-de-I’Etanson, 77270 Nangis, France, folia. The only report on A. columnaris is a 12, pp. 49-55 preliminary study using seedling stem Burrows G.E. (1983) Organ culture of some segments (Burrows, 1983). In our study, species of Araucariaceae. Anatomical aspects tips of very small branchlets appearing of bud development. Proc. 2nd Aust. Plant Tis- naturally on plagiotropic branches of 14 yr sue Cult. Conf. S’ydney. pp. 18 old trees were used. In nature, these Gupta P.K. (1987) Advances in biotechnology of branchlets hardly show any growth. In cul- conifers. Curr. Sci. 57, 629-637 ture, however, these shoots appeared to Haines R.J. & de Fossard R.A. (1977) Propaga- show normal orthotropic growth. In tion of hoop pine (Araucaria cunninghamii Ait). Sequoia, Boulay (1979) also obtained Acta Hortic. 78, 297-302 orthotropic behavior of in vitro shoots Handro W. (198E;) Araucaria. In: Biotechnology derived from stump sprouts of mature in Agriculture & f=orestry (Bajaj P.S., ed.), Vol 1, trees. Explants from mature trees of some Trees, Springer-Verlag, Berlin, pp. 310-315 5 other conifers have also been used Maene L. & Debergh P. (1987) Araucaria. In: (Bonga, 1981; Gupta, 1987). Cell and Tissue Culture in Forestry. (Bonga J.M. & Durzan D.J., eds.), Vol. 3, Cell The present studies also show that sea- Histories, Gymnosperms, Angiosperms and and prechilling of explants effect sub- son Palms. Martinus Nijhoff, Dordrecht, pp. 176-184 sequent response in culture. Thiourea T. & Skoog F. (1962) A revised Murashige seems to promote axillary branches and medium for rapid growth and bioassays with shoot growth, as observed by Maene and tobacco tissue cultures. Physiol. Plant. 15, 473- 497 Debergh (1987).

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