Indução de brotações em Phalaenopsis cultivada in vitro com radiação UV e padronização das análises moleculares por SSR

Abstract

Although orchids show exuberante beauty, the economic sector seeks to increase the genetic variability of some species of comercial interest, such as Phalaenopsis, to meet Market demand. A viable alternative to broadening phenotypes is to acquire genetic variability through induced mutation that alters the orchid’s genome sequence, which can be accomplished by exposure to ultraviolet light (UV). UV light are an efficient mutagenic agent because its energy is able to excite electrons and its efficiency is related to the exposure time of the sample. Therefore, the aim presente work is to induce random mutation and get sprouts in Phalaenopsis hybridum exposed to UV light for different periods to generate new alleles. For, this, an in vitro culture system was used, comparing two culture media with diferente amounts of explants. To establish the efficiency of the culture medium parameters such as number of roots, number of shoots, length of the longest root, shoot height and number of leaves were measured. The results indicated that the suitable culture medium for in vitro cultivation of hybrid Phalaenopsis is Knudson médium supplemented with 10 %(v/v) carrot juice, since there was a lower incidence of contamination and oxidation of the cultivated plants. In order to obtain sprouts, besides the carrote juice medium, it was verified that the exposure time to UV light of 30 minutes was more effective. DNA extractions and Polimerase Chain Reaction (PCR) reactions were successfully performed, establishing eficiente protocol for hybrid Phalaenopsis. No mutations were evidenced by phenotype analysis of hybrid Phalaenopsis, as they are Young seedlings.