Micropropagation of columnaris Hook L. Sehgal, O.P. Sehgal, P.K. Khosla

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L. Sehgal, O.P. Sehgal, P.K. Khosla. Micropropagation of Araucaria columnaris Hook. Annales des sciences forestières, INRA/EDP Sciences, 1989, 46 (Suppl), pp.158s-160s. ￿hal-00882526￿

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L. Sehgal* O.P. Sehgal P.K. Khosla

DYSP University of Horticulture and Forestry, Solan 173 230,

Introduction regulators and/or other chemicals. The cultures were kept at 25°C under cool white fluorescent light (1.5 klux), from 40 W tubes, for 16 h each The genus Araucaria comprises several day. At least 20 replications were used for each treatment. magnificent evergreen forest tree species, including Araucaria columnaris Hook., which has a high ornamental value as well. Conventional seeds propagation by Results is slow and inadequate for producing large uniform progenies. Cuttings are difficult to root and they produce plagiotropic 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 NM), were est:ablished on the medium 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 was obtained by subculturing on a medium containing 6-benzylaminopurine Tips (3-5 mm) of secondary and tertiary lateral (BAP, 12.5 pM), indole butyric acid (IBA, 1 branches, taken from 14 yr old trees, were steri- !M) and 3% sucrose. lized in mercuric chloride for 1 min and (0.05%) taken in for cultured on Murashige and Skoog’s (MS) Explants January, prechilled medium (Murashige and Skoog, 1962), contain- 1 mo at 5°C and cultured on MS medium ing different concentrations of various growth (containing 10 uM kinetin and 1 !M IBA)

* Permanent address: Gargi College, Delhi University, N. Delhi 110049, India. produced almost twice the shoot growth Subculturing or direct planting on a rate as compared to the ones not chilled, medium containing kinetin (10 pM) and giving a shoot length of 1.5 cm in 6 wk thiourea (1.3 ,uM) enhanced the shoot (Table I). length and production of axillary branches (Fig. 1 The latter were separated and Araucaria is a hard-to-root material in used for multiplication. Addition of 0.3% culture, and limited success in rooting has 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 production of shoot growth and axillary branches obtained in culture in the pres- Discussion and Conclusion ent study is an important step in micro- propagation. Attempts to induce in vitro are in Micropropagation of most of the rooting progress. 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, 1977; Maene & Debergh, 1987), only Bur- References 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- sues from mature conifers. In vitro 17, 511-5188 ly, Handro (1986) reported the develop- Boulay M. (1979) Multiplication et clonage ra- ment of shoot in from axillary vitro, starting pide du Sequoia sempervirens par la culture in branches of a 20 yr old tree of A. angusti- vitro. In: Etudes et Recherches, AFOCEL, folia. The only report on A. columnaris is a Domaine-de-I’Etanson, 77270 Nangis, France, preliminary study using seedling stem 12, pp. 49-55 segments (Burrows, 1983). In our study, Burrows G.E. (1983) Organ culture of some of . Anatomical tips of very small branchlets appearing species aspects of bud development. Proc. 2nd Aust. Tis- on branches of 14 naturally plagiotropic yr sue Cult. Conf. S’ydney. pp. 188 old trees were used. In nature, these P.K. Advances in of branchlets show In cul- Gupta (1987) biotechnology hardly any growth. conifers. Curr. Sci. 57, 629-637 ture, however, these shoots appeared to Haines R.J. & de Fossard R.A. show normal In (1977) Propaga- orthotropic growth. tion of hoop pine (Araucaria cunninghamii Ait). Sequoia, Boulay (1979) also obtained Acta Hortic. 78, 297-302 behavior of in vitro shoots orthotropic 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. 310-3155 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 Vol. 3, Cell The studies also show that sea- D.J., eds.), present Histories, Gymnosperms, Angiosperms and son and of effect sub- prechilling explants Palms. Martinus Nijhoff, Dordrecht, pp. 176-184 sequent in culture. Thiourea response Murashige T. & Skoog F. (1962) A revised 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- Debergh (1987). 497