Figure 1.
Flowchart showing the major steps for chloroplast isolation according to high salt plus saline Percoll.
Table 1.
Composition of chloroplast isolation buffers and wash buffers for modified high salt method, high salt plus saline Percoll method and sucrose gradient method.
Figure 2.
Chloroplast visualization of Araucaria angustifolia in phase contrast microscopy.
(A) Chloroplasts isolated with improved high salt method; (B) Chloroplasts isolated with sucrose method; (C) Chloroplasts isolated with high salt plus Percoll method; (D–F) Micrographs during chloroplast isolation with high salt plus saline Percoll method; (D) Broken and intact chloroplasts before Percoll gradient centrifugation; (E) Intact isolated chloroplasts in interface 70/30% after Percoll gradient centrifugation; (F) Broken chloroplasts in upper 30% phase after Percoll gradient centrifugation. Dotted arrows indicate broken chloroplasts. Solid arrows indicate intact chloroplasts. Bar –50 µM.
Figure 3.
Chloroplast DNA visualization of Araucaria angustifolia in 0.7% agarose gel stained with ethidium bromide.
(A) Ladder 1 kb and cpDNA isolated with modified high salt method; (B) Ladder 1 kb and cpDNA isolated with sucrose method; (C) Ladder 1 kb and cpDNA isolated with high salt plus saline Percoll method.
Table 2.
cpDNA from different isolation methods in Araucaria angustifolia sample.
Table 3.
cpDNA ratios of selected conifers evaluated using Nanodrop®.
Figure 4.
Reference graph track showing observed coverage values.
Different colors show the minimum (light blue), mean (blue), and maximum (dark blue) observed coverage values for all genomic regions (data aggregation above 100 bp). Araucaria angustifolia, Araucaria bidwilli, and Podocarpus lambertii sequence reads were mapped on Podocarpus totara; Pinus patula sequence reads were mapped on Pinus thunbergii.
Table 4.
Average coverage of cpDNA evaluated from selected conifers with CLC Genomics Workbench 5.5 software.