Figure 1.
Isolation of two-celled proembryo, apical, and basal cells.
(A) Isolated two-celled proembryo; (B) Apical and basal cell protoplasts; (C) Separation of apical and basal cell protoplasts; (D) Two-celled proembryo for laser ablation; (E) Apical cell ablation of basal cell; (F) Basal cell after ablation of apical cell. Bar = 20 µm.
Figure 2.
Apical and basal cell expressed sequence tag (EST) clusters compared to zygote EST clusters.
EST cluster comparison between apical (Ap)/basal cell (Ba) ESTs and a zygote.
Table 1.
Most abundant clusters in tobacco apical cells.
Table 2.
Most abundant clusters in tobacco basal cells.
Figure 3.
Functional cluster categories and expressed sequence tags from apical and basal cells.
A: comparison of clusters between apical and basal cell. B: comparison of ESTs between apical and basal cell. Ap, apical cell; Ba, basal cell.
Figure 4.
RT-PCR examination of transcripts in apical and basal cells.
Of 71 expressed sequence tags (ESTs) examined by RT-PCR, only ntb1051 and ntb0886 could be confirmed exclusively in basal cells. The transcripts of five genes, nta0801, nat0809, ntb1147, ntb1892, and nta2449, were accumulated at a higher level in apical cells, whereas transcripts of three genes, nta0893, nta1009, and ntb0946, were accumulated more in basal cells.
Figure 5.
Transcript levels of selected genes in apical/basal cells as shown by real-time RT-PCR.
(A, B): The expressed sequence tags (ESTs) showed specific distribution in apical cell. (C–O): ESTs showed a significantly higher level in apical vs. basal cells (≥2-fold difference). (P–S): ESTs showed a significantly higher level in basal vs. apical cells (≥2-fold difference). Ap, apical cell; Ba, basal cell; Zy, zygote. Expression level in apical cell was set to 1.
Figure 6.
qPCR results showing expressed sequence tags (ESTs) existence in zygote cells.
Of the 42 transcripts examined, 39 ESTs were found in zygote.