Application of balloon type bubble bioreactor for micropropagation of Aloe barbadensis and Lilium hybrid
Author(s)
Cardona Suarez, Carolina Maria
Date Issued
November 25, 2013
Type
Doctoral Thesis
Abstract
The objective of the research was to improve the application of balloon type bubble bioreactor for
plant micropropagation of Lilium hybrid and Aloe barbadensis. Bioreactor provides optimum
growth conditions by regulating chemical or physical parameters to achieve either maximum yield
and high quality of the propagules, or to keep the production costs as low as possible by integration
of automated facilities and simple low-cost devices. The first experiment was performed in order to
describe a method for micropropagation of pollenless Asiatic hybrid lilies taking into consideration
the genotype, the culture medium and the scale size.
A plant micropropagation protocol for Lilium using an automated balloon type bubble bioreactor
(BTBB) was set up in a second experiment. Bulblets of Lilium (LA) hybrid “Brindisi” were grown
in two different culture methods: continuous immersion and temporary immersion (ebb and flood
system) in liquid medium. Results revealed that morphological traits and biomass accumulation
were more efficient when bulblets were cultured in continuous immersion bioreactor. In addition it
was found a technique to reduce the risk of contamination during bioreactor culture. This technique
includes the sterilization of Lilium clusters with sodium hypochlorite for 5-15 minutes before
setting up the bioreactor and the use of cefotaxime (0.50 mg ml -1) in the liquid medium.
Even it was examined the effect of ozone sterilization in bioreactor culture of Lilium. Moreover it
was examined the dynamic of Lilium bulblet growth for the optimum culture period. Results
indicated that the use of ozone in bioreactor culture of Lilium is an efficient instrument to eliminate
the high risk of contamination in the liquid substrate without the use of sophisticated technology
and with optimal morphological parameters of growth. Only low signs of oxidative stress were
observed in bulblets treated with ozone. The dynamic of bulblet growth revealed that cultured
Lilium for 30 days in 4 l balloon type bubble bioreactor system was the optimum culture period.
The influence of ozone treatments on in vitro propagation of Aloe barbadensis in continuous
immersion bioreactor was studied. Results provide experimental evidence that the use of ozono
inside the bioreactor for the micropropagation of A. barbadensis is a viable and efficient model for
automation and large-scale production of quality shoots.
Additional information
Dottorato di ricerca in Ortoflorofrutticoltura
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