Protocols for Micropropagation of Woody Trees and Fruits

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B. PINTOS ET AL.
paper. Then the explants were subcultured on SM medium supplemented with 3%
(w/v) sucrose, 500 mg.l–1 glutamine and solidified with 0.8% (w/v) agar, pH = 5.6.
The explants were viable when survived the treatment and maintained the embryogenic
capacity. In the control, survival reached the 96%, while oryzalin 0.01 mM
provided at least 78.25% viability. Flow cytometry analysis of haploid embryos treated
with oryzalin showed that while control embryos remained haploid, 46.7% oryzalintreated
embryos became diploid (Figure 4) (Pintos, 2005).
a
BEST
DIPLOIDIZATION
TREATMENT: 0.01 mM
ORYZALIN for 48
HOURS
≈ 50.00% DIPLOID
EMBRYOS (induced
Doubled-Haploids)
89.03% HAPLOID EMBRYOS
c
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count
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400
Figure 4. Induction of cork oak doubled-haploids through 0.01 mM oryzalin application.
A) Initial haploid embryos. B) Flow cytometry histogram of relative DNA content of nuclei
released from anther derived cork oak embryos prior to diploidization treatment. C) oryzalintreated
embryos. D) Flow cytometry histogram of relative DNA content of nuclei released from
diploidized cork oak embryos after oryzalin treatment.
100
0
0 50 100 150 200 250 300 350 400 450 FL1500 partec
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count
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b
d
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