Comparative study on the in vitro multiplication potential of Magnolia ...

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“FACULTATEA DE HORTICULTURĂ ŞI SILVICULTURĂ”material was transferred in a solution of calciumhypochlorite concentration of 6% for another 20minutes – treatment applied on vegetating buds. Forthe buds in the foliation stage as well as for the apicalbuds during full growth, the treatment was reduced tohalf of the initial time.The culture media were sterilized byautoclaving at 120°C temperature for 20 minutes.Before autoclaving, the pH registered in a culturemedium was adjusted to 5.6-5.8.The surgical type instruments used weresterilized in the drying stove, at 120 o C temperature for2 hours.For the growing, multiplication and rooting ofexplants we ensured in the growing room controlledconditions of temperature (22-24 o C), photoperiod (16hours) and light intensity.Composition of nutritive media used for culture initiationGrowth regulators used and theirVariconcentration in culture medium (mg/l)antsBasic mediumexperimentalascorbicBAP IAA NAA GA 3acidV.1 Macroelements MS, 1 0,2 - - -V.2 microelements MS, vitamins JQ 0,5 - 1 0,1 5V.3 Macroelements MS, 1 0,2 - - -V.4 microelements MS, vitamins MS 0,5 - 1 0,1 5V.51 0,2 - - -Macroelements MS,V.6 0,5 - 1 0,1 5microelements MS, vitamins LSV.7 0,7 - 1 0,1 5Legend: MS = Murashige - Skoog (1962); LS = Linsmaier – Skoog (1965); JQ = Jaquoit (1956)VariantsexperimentalComposition of nutritive media used for micropropagationBasic mediumGrowth regulators used and theirconcentration in culture medium (mg/l)BAP 2iP TDZ Kin0,25 - - -V.1V.2 0,5 - - -V.3V.4V.5Macroelements MS,microelements MS,vitamins M---0,250,5---0,25---V.6 - - 0,5 -V.7 - - - 0,25V.8 - - - 0,5Legend: MS = Murashige - Skoog (1962); M = Miller şi colab. (1982)AComposition of nutritive media used for in vitro rooting of shootsGrowth regulators used and theirVariantsBasic medium concentration in culture medium (mg/l)experimentalIBA GA 3V.1- 0,1V.2 2 0,1V.3 Macroelements MS½ n,4 0,1V.4 microelements MS½ n,6 0,1V.5 vitamins LS n8 0,1V.6 10 0,1V.7 12 0,1V.8 14 0,1Legend: MS = Murashige - Skoog (1962); LS = Linsmaier – Skoog (1965)Table 1Table 2Table 3The achieved results was registered asmultiplication rate being express inmicroshoots/explants and was statistic interpretation byDuncan Test. Growing of explants and rooting ofmicrocuttings was express in percents.40
Results ObtainedThe initiation phaseThe reactivity of different types of explants onvarious culture media was tested in order to establishexperimental protocol for in vitro multiplication ofmagnolia species.In the initiation stage of in vitro culture, thegrowing of explants was influenced by the type ofexplants and composition of nutritive media.Results obtained have shown that explants ofapical buds represent optimal source of inoculums.The period most indicated for sampling andinoculating explants is November-December whenvegetative buds are in the dormant stage. Research inthe evaluation of morphogenetic capacity of explantsshowed that nutrient media composition influencedreactivity of explants taken during active growth ofbuds similar with explants taken from dormant stage,differences consisting of percentages of explants turnedevolving, which were lower.It was found that the best results in terms ofpercentage of explants started the trend was obtainedon the version supplemented with 0.7 mg/l BAP, 1mg/l NAA, 0.1 mg/l GA 3 , 5 mg/l ascorbic acid andvitamins LS (V.7), followed the versions V6, V3, V5,V4, V1 and V2.In general, nutrient media compositioninfluenced the reactivity of explants taken of axillarybuds similar with explants taken from apical buds,differences consisting of percentage of explants startedevolving, that were smaller (fig. 1).apical budaxillary budFig. 1. The in vitro reactivity of magnolia species considering the explant type, genotype and culture mediumcompositionFig. 2. Morphogenetic response of explants, 4 weeks after initiation of in vitro cultureThe multiplication phaseIn multiplication phase, the analysis of theeffect of type and concentration of citokinine could beseen the clearly influence of citokinine type on theregeneration rate.In the case of Magnolia stellata species,referring to the effect of citokinine depending on its41
Page 1: Volume 16(2), 39-44, 2012JOURNAL of
Page 5 and 6: Fig. 5. The influence of indolilbut

Comparative study on the in vitro multiplication potential of Magnolia ...

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