Flower buds were harvested, transported under melting
ice, surface sterilized and immediately dissected.Before
plating the anthers, bud and anther lengths were
systematically measured with a caliper. Only those of
the optimal length were considered for anther culture.
A variable number of anthers (ranging from 68 to 225
per accession) were plated. Anthers were cultured
according to Dumas de Vaulx and Chambonnet (1982).
Briefly, anthers were inoculated in dishes with agarbased
Ct inductive medium and cultured for 8 days in
darkness at 35C. Then, they were transferred to light
(12/12 photoperiod) and 25C for four more days. At
day 12, anthers were transferred to R1 medium, where
they were cultured indefinitely at 25C, refreshing
medium every 20 days. Once embryos emerged from
anthers, they were isolated and cultured individually on
dishes or pots with V3 medium for germination and
transformation into plantlets.
Flower buds were harvested, transported under meltingice, surface sterilized and immediately dissected.Beforeplating the anthers, bud and anther lengths weresystematically measured with a caliper. Only those ofthe optimal length were considered for anther culture.A variable number of anthers (ranging from 68 to 225per accession) were plated. Anthers were culturedaccording to Dumas de Vaulx and Chambonnet (1982).Briefly, anthers were inoculated in dishes with agarbasedCt inductive medium and cultured for 8 days indarkness at 35C. Then, they were transferred to light(12/12 photoperiod) and 25C for four more days. Atday 12, anthers were transferred to R1 medium, wherethey were cultured indefinitely at 25C, refreshingmedium every 20 days. Once embryos emerged fromanthers, they were isolated and cultured individually ondishes or pots with V3 medium for germination andtransformation into plantlets.
การแปล กรุณารอสักครู่..