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Master's Dissertation
Full name
Maria Cristina Falco
Knowledge Area
Date of Defense
Piracicaba, 1993
Title in Portuguese
Cultura de células em suspensão e protoplastos em cana-de-açúcar (Saccharum spp)
Keywords in Portuguese
Abstract in Portuguese
Para possibilitar a utilização de técnicas de biologia celular e molecular em variedades nacionais de cana-de-açúcar foram selecionados calos a partir de explantes foliares que deram origem a suspensões celulares embriogênicas. Tais suspensões tiveram seu crescimento quantificado segundo parâmetros como número de células, peso fresco, peso seco e volume do total de células. Plantas foram regeneradas a partir de calos e suspensões celulares com até cinco meses de subcultivo. Protoplastos foram isolados a partir de calos e células em suspensão em diferentes soluções enzimáticas, métodos e idades de cultura, calos foram obtidos de protoplastos isolados de células em suspensão, mas não houve regeneração de plantas
Title in English
Cell suspension culture and protoplast in sugarcane (Saccharum spp)
Abstract in English
This work was carried out to study callus induction, ceel suspension initiation and protoplast isolation and culture. Two cultivated sugarcane varieties were used namely SP 70-1143 and SP 79-1011. The following steps were accomplished: (1) Selection of the best culture conditions to promote callus formation and support the next steps. (2) Cell suspension culture were iniciated and cells selected which would had cells with proper morphogenic characteristics to be used in protoplast isolation and plant regeneration. The selection of the best technique was supported by parameters obtained from number of cells, fresh and dry weigth, and packed cell volume. The variable number of cells was the best expression to the cellular suspension growth. Different culture media were evaluated with emphasis on the 2, 4-D dosages requerid for long time duration of the cell culture. The MS culture medium with 1.5 to 3.0 mg / 1 of 2,4-D, 500 mg / 1 of hidrolysed casein and coconut milk (5 %) showed efficient. ( 3) Plant regeneration from cell suspensions were tested to establish the conditions for the use of biobalistic techniques. The MS culture medium was able to regenerate plants in a cellular suspension until the period of five months. (4) Protoplast isolation were studied using different enzymes with known methodologies and different culture age. The rates of isolation, colony and microcallus formation were recorded. The enzyme solution containing Cellulase RS (1 %), Pectolyase Y23 (0.2 %), EMS (5.0 mM) and CPW salts with manitol (13 %) as osmoticum yielded a good number of protoplasts. Calli derived from protoplasts were obtained in three differents media, being R2 the best followed by N6 and GM. ( 5) Plant regeneration from protoplasts. The work is in progress but no plant has been regenerated
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