S04 - Session O6 - Optimization of in vitro cultivation of Cattleya nobilior Rchb.f in a temporary immersion bioreactor EMBRAPA/C4 Científica

Authors: Mônica Rosa Bertão *, Vinícius Manoel Bertão, Camila Almeida Pimentel, Milena Cristina Moraes, Larissa Frigui Moreira , Carlos Alberto de Lima Conte, Darío Abel Palmieri

Orchid cultivation of the genus Cattleya , denotes a great importance activity in world agribusiness. Cattleya nobilior Rchb.f high ornamental value, draws attention. However, because the specie occurs in areas of great anthropic interference and exhibit a high risk of indiscriminate collection, its habitat is vulnerable, which can lead this species to a threatened category. One perspective to positively influence micropropagation is the use of temporary immersion bioreactors. In this work, the Temporary Immersion Bioreactor system (TIBs - EMBRAPA/C4 Cientifica) was applied, evaluating the use of the biocide Kathon®CG and sanitizing solution of sodium hypochlorite. MSP09 culture medium supplemented with 40.0 g.L -1 of sucrose (pH 5.8) and doses of 120, 240, 360 and 480 uL/L of Kathon®CG were assessed. As control treatment, the medium was autoclaved at 121ºC and 1 atm (20 minutes). A total of 20 seedlings (1.5 cm high) / vial were placed in the TIBs. Vials, accessories and utensils from the TIBs were sterilized with 0.003% NaClO and the media prepared with 0.0005% NaClO. No contaminations were observed in the flasks with the different doses of Kathon®CG, over 82 days of culture. Autoclaving was not effective for medium sterilization, contaminating on the 7th day of culture. The media with Kathon®CG showed 100% efficiency in contamination control, without explants phytotoxicity. The use of NaClO was efficient in sterilizing the culture medium and sterilizing the TIB system. Seedling growth through fresh weight showed that the seedling weight increased between 12.22 and 17.65 times. The TIBs allowed the significant sprouting of the material, going from 80 initial explants to 2260 sprouts. The results demonstrate an optimized protocol for C. nobilior cultivation in an EMBRAPA/C4 Cientifica Temporary Immersion Bioreactor system. suggested: The cultivation of the genus Cattleya represents an important economic value. The high ornamental value of Cattleya nobilior attracts attention so that it is collected indiscriminately. However, its habitat is fragile, so this species may end up in an endangered category. In this study, it was propagated in temporary immersion bioreactors (TIBs - EMBRAPA/C4 Cientifica), evaluating the use of the biocide Kathon®CG and a disinfecting solution of sodium hypochlorite. MSP09 culture medium supplemented with 40.0 g.L-1 sucrose (pH 5.8) and doses of 120, 240, 360 and 480 uL/L of Kathon®CG were evaluated. A total of 20 seedlings (1.5 cm high)/vial were placed in the TIBs. Vials, accessories and utensils of the TIBs were sterilized with 0.003% NaClO and the media was prepared with 0.0005% NaClO. No contaminations were observed in the flasks containing the different doses of Kathon®CG, during 82 days of culture. As a control treatment, the medium was autoclaved at 121ºC and 1 atm (20 minutes). Autoclaving was not effective for sterilization of the medium, with contamination on the 7th day of culture. The media with Kathon®CG showed 100% efficiency in controlling contamination, with no phytotoxicity of the explants. The use of NaClO was efficient for sterilization of the culture medium and sterilization of the TIB system. Seedling growth by fresh weight showed that the seedling weight increased between 12.22 and 17.65 times. The TIBs allowed significant sprouting of the material, ranging from 80 initial explants to 2260 shoots. The results demonstrate an optimized protocol for the culture of C. nobilior in an EMBRAPA/C4 Cientifica temporary immersion bioreactor system.