Online Resource 3. Effect of sucrose concentration and embryo proliferation media on the mean number of nodular embryogenic structures (NS) and secondary somatic embryos (SSEs) generated by torpedo-stage embryos used as subcultured explants for secondary embryogenesis in Q. rubra. Proliferation medium consisted of STS-basal medium (BM) with or without activated charcoal (AC) or supplemented with 0.44 µM BA.

Sucrose
concentration
(%) / Proliferation media / Total Number NS ± SE / Total Number SSEs ± SE / Developmental stage of SSEs
1Heart-torpedo No. ± SE / 1Cotyledonary ≤ 5mm No. ± SE / 1Cotyledonary > 5mm No. ± SE
3 / BM + BA / 15.8 ± 5.9 / 11.5 ± 2.1 / 4.0 ± 0.6 / 4.3 ± 0.9 / 3.5 ± 1.3
3 / BM / 18.8 ± 9.6 / 14.5 ± 1.0 / 4.5 ± 1.7 / 5.8 ± 2.1 / 4.3 ± 1.0
3 / BM + AC / 31.5 ± 5.6 / 24.3 ± 4.4 / 17.3 ± 2.3 / 2.3 ± 1.3 / 4.5 ± 1.4
6 / BM + BA / 6.0 ± 1.9 / 6.5 ± 1.3 / 2.3 ± 1.6 / 1.5 ± 0.3 / 2.8 ± 0.5
6 / BM / 19.0 ± 4.2 / 14.8 ± 2.1 / 7.0 ± 1.8 / 4.0 ± 0.4 / 3.8 ± 0.9
6 / BM + AC / 56.8 ± 16.5 / 24.0 ± 7.9 / 20.3 ± 5.2 / 3.8 ± 1.7 / 0.0 ± 0.0
F-test
Proliferation media (A) / P≤0.01 / P≤0.01 / P≤0.001 / ns / ns
Sucrose concentration (B) / ns / ns / ns / ns / P≤0.05
AxB / ns / ns / ns / ns / ns

1Mean number of secondary embryos at specific developmental stage produced per Petri dish.

Each value represents the mean ± standard error (SE) of four replications with ten explants in each replicate.

Statistical significance (P-values) of the terms of a two-way ANOVA with proliferation media and sucrose concentration as the two main factors. ns, not significant.