Abstract. Pitoyo A, Hani MR, Anggarwulan E. 2015. Application of chitosan spraying on acclimatization success of tiger orchid (Grammatophyllum scriptum) plantlets. Nusantara . In Indonesia, tiger orchid or anggrek macan (Grammatophyllum scriptum) is one of the orchid species that have been mass propagated with in vitro culture technique. The successful determinant of in vitro culture after planlet formation is acclimatization. Plantlet have faced with absolutely different environmental condition between inside and outside bottle that potentially severe the survival of the plantlet. The objectives of this research were to know the effect of chitosan for success of chitosan spraying in acclimatization of plantlets of G. scriptum and to know the optimum concentration of chitosan that can help the successful of the acclimatization of plantlets G. scriptum. This research used a Completely Randomized Design (CRD) with four level concentrations of chitosan spraying, there were 0 g/L; 0,5 g/L; 0,75 g/L; 1 g/L. Every treatment underwent 15 repetitions. The data taken on this research were qualitative and quantitative data. Qualitative data was got from observation of morphology and anatomy then it was analyzed descriptively. The quantitative data was got from percentage of plantlets life, observation of shoot length, leaves length, leaves width, number of leaves, number of shoot, index and density of stomatal. The quantitative data was analyzed by Analysis of Variance (ANOVA), if there was a significant difference between treatment groups then followed by Duncan's Multiple Range Test (DMRT) at level significance of 5 %. The result of this observation indicated that the effect of chitosan was significant on several parameters including width leaf, plantlets height, and stomatal index while the length and number of leaves were not significantly different. The concentration of chitosan real effected on these ranged from 0.5-0.75 g/L. Chitosan concentration at above of 0,75 g/L showed a negative influence on the acclimatization successfully of plantlets G. scriptum.
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