Figure 1.
Crossing scheme to generate a fluorescence-tagged line harboring a heterozygous mutation in the quartet1 (qrt1) mutant background.
The mutation of interest depicted here is a generic mutation in a subunit of translation initiation factor 3 (eIF3x).
Figure 2.
Fluorescence-tagged pollen grains prior to germination.
(A) Mature eif3h-1 mutant pollen grains (unmarked) in comparison with wild type (marked with YFP). In the middle panel one mutant pollen grain has germinated prematurely. (B) Mature eif3e-1 mutant pollen grains (unmarked) in comparison with wild-type pollen (marked with DsRed). The genotypes of the parent plants are indicated to the left of each panel.
Figure 3.
Pollen grains prior to germination.
(A–C) Mature pollen grains were stained with Alexander's stain to check for viability. (A) wild type, (B) eif3h-1, and (C) eif3e-1 mutant. (D–F) Mature pollen tetrads were stained with DAPI to visualize the nuclei. (D) wild type, (E) eif3h-1, and (F) eif3e-1 mutant. The location of the vegetative nucleus (dotted outline) was confirmed by refocusing when it was not in the same plane as the two sperm nuclei. Genotypes of the anther bearing plants are indicated.
Figure 4.
Germination of eif3h and eif3e mutant pollen.
(A, B) While pollen germination of eif3h-1 pollen (non-fluorescent) was low (A, also see Table 1), such pollen grains did occasionally germinate (B, arrow). (C, D) Similar results were obtained for eIF3e. The tetrad shown (C) clearly demonstrates that at least one of two eif3e-1 mutant pollen has germinated. Generally, however, eif3e-1 pollen germinates poorly, as seen in the representative tetrad below (D). Note that, in the long exposures needed to visualize fluorescence from pollen tubes, the bright fluorescence of the marked pollen grains often reflects from the cell wall of unmarked pollen grains.
Figure 5.
Pollen tube lengths in eif3h and eif3e mutants.
Pollen tube lengths were measured for one hundred non-fluorescent germinated pollen grains of (A) eif3h-1 and (B) eif3e-1 and compared with one hundred fluorescent (bona fide wild-type) pollen tubes. Data from two independent replicates were pooled. The difference in mean length was significant for eif3h (P<10−6, t-test), but not for eif3e (P>0.05).
Table 1.
Germination of pollen from eIF3h+; FTL567/eif3h−; + heterozygous parents.
Table 2.
Germination of pollen from eIF3e+; FTL1413/eif3e−; + heterozygous parents.