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Báo cáo khoa học: "Micropropagation of Eucalyptus viminalis Labill"

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viminalis Labill Micropropagation of Eucalyptus 3 Araújo 2 Graga A.J. de M.E. Cortezzi M. Wiecheteck 1 UFPR, Pisa Florestal S.A., Rod. PR-151,km 232, 84200, JaguariaivalPR, 2 CNPF, Centro Nacional de Pesquisa de FlorestaslEmbrapa, C.P. 3319, 80001,CuritibalPR, and 3 UFPR, School of Forestry, C. P. 2959, 80001,CuritibalPR, Brazil Materials and Methods Introduction Nodal segments (about 1 cm long) from green- viminalis has been established Eucalyptus house-grown E. viminalis seedlings were in southern Brazil mainly because of its washed under running water for 1 h, soaked in frost tolerance and growth potential. a 10% (vlv) commercial detergent for 20 min, However, its slow growth rate and undesir- and afterwards in a 100 ml solution of 1% N with 2 drops of Tween 20 for 10 min. CIO A able stem form, due mainly to a narrow The disinfectants were removed by 3 succes- genetic base, have limited its extensive sive rinses in autoclaved, distilled water. use. Vegetative propagation of selected Explants were grown on half-strength MS genotypes of this species has not been medium (Murashige and Skoog, 1962) sup- plemented with NAA and BAP, both at 0.1 very encouraging, the major constraint mg 2% sucrose, 0.8% Difco Bacto agar and , 1 I- ’ being a low percentage of rooted cuttings vitamins as described by Gamborg and Wetter (Cunningham and Mott, 1985). Consid- (1975). At the multiplication phase, different erable effort has been exerted to develop levels of NAA (0, 0.1, 0.5, 1.0 or 1.5 mg!l-t) and BAP (0, 0.05, 0.1, 0.2, 0.5 or 1.0 mg were ) 1 I- ’ in vitro techniques for rapid clonal propa- added to the MS medium. In the shoot elon- gation and for improvement of Eucalyp- gation experiment, IBA (0 or 1.0 mg-1- and ) 1 tus. Recent studies have shown that mul- GA (0, 0.1 or 1.0 mg!l-t) with or without 3 tiple axillary buds were obtained by using activated charcoal (A.C.) (15 mg were used ) 1 I- ’ in MS medium. Elongated shoots were subcul- in vitro techniques of juvenile E. viminalis tured onto an MS or half-strength MS medium (Mehra Palta, 1982; Cunningham and with different levels of IBA (0.1, 0.5 or 1.0 Mott, 1985). However, plant regeneration mg!l-!) and KIN (0 or 0.1 mg!l-!) to initiate low due to poor rhizogenesis (Mehra roots. The pH of the medium was adjusted to was 5.8. The experiments were carried out in a Palta, 1982). This paper describes a more randomized complete block design, each treat- successful system for micropropagation of ment being repeated 15- 30 times depending E. viminalis. upon the phase. Cultures were maintained Abbreviations: NAA: naphthalene acetic acid; BAP: benzylaminopurine; IBA: indole butyric acid, cid: GAg: gibberellic b en zylaminopurine; ’cid acid 3; KIN: kinetin.
shoot proliferation. This was confirmed 16/8 h and under light/dark photoperiod a temperature of 25+2°C. After the rooting with 0.1 mg which gave the highest , 1 I- ’ transferred to shade- phase, plantlets a were multiplication rate (6.2 shoots/explant). A house. decreasing trend of the rate of multiplica- tion was observed when levels of BAP were reduced below 0.1 mg (Fig. 2). 1 I- ’ Results the use of During the elongation phase, 0.1 mg!l-! g of A.C. with or without 1 I- ’ 15 of IBA in MS medium resulted in the most Optimal shoot proliferation was achieved elongated shoots (Fig. 3). Shoots grown by adding 0.2 mg of BAP and 0.1 1 I- ’ on medium containing A.C. only showed 1 I- ’ mg of NAA (4.6 shoots/explant) (Fig. the true morphological characteristics of 1As the concentration of BAP increased, the species. The use of GA was found to 3 shoot formation decreased. Although the be detrimental to elongation of E. vimina- multiplication rate was higher with 0.5 or lis shoots. 1.0 mg of NAA, the lowest concentra- 1 I- ’ tion of NAA (0.1 mg!l-!) provided more The elongated shoots produced more vigorous shoots than the other treatments. half-strength MS medium with 0.5 roots on or 1.0 mg of IBA, although better root 1 I- ’ The fact that lower levels of BAP gave morphology was observed with 0.5 mg!l-! higher multiplication rates suggested that levels below 0.2 mg could increase 1 I- ’ of IBA (Table 1).
Discussion and Conclusion Acknowledgmients The authors are grateful to Prof. Dr. Mario successful system for the micro- A more Takao Inoue and Prof. Dr. Flavio Zanette for propagation of E. viminalis was developed their assistance and Mr. Adhemar Villela Filho, in this study. Forest Director of Pisa Florestal S.A., for encouragement and support throughout this Multiplication rates of 5-6 shoots/ex- study. plant were obtained by using 0.2 mg of 1 I- ’ BAP and 0.1 mg of NAA on MS 1 I- ’ medium. Although these are higher than References those reported to date for E. viminalis, they are lower in comparison to other Eucalyptus species. Shoot elongation of Cunningham M.tN. & Mott R.L. (1985) Micropro- pagation of Eucalyptus viminalis. Proceedings E. viminatis was not improved by using of the i8th Southern Forest Tree Improve- GA Results were opposite to those . 3 ment Conference. Long Beach, Mississippi. pp. expected, since the elongation effect 54-63 should be one of the main features of O.L. 8! Wetter L.R. (1975) In: Plant Gamborg GA The best root formation (66.6% of . 3 Tissue Culture Methods. National Research Council of Canada, Saskatoon, pp. 109, p. 91.1 .I rooting) was obtained on a half-strength Mehra Palta A. (1982) Clonal propagation of MS medium, supplemented with 0.5 Eucalyptus by tissue culture. Plant Sci. Lett. 26, mg.!- of IBA. This study establishes a 1 1-11 1 micropropagation method for juvenile E. T. & Skoog F. (1962) A revised Murashige viminalis which can serve as a basis for medium for rapid growth and bioassays with the in vitro propagation of adult trees for tobacco tissue cultures. Physiol. Plant. 15, 473- future improvement programs. 497

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