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. 2021 Jul 27;12:704958. doi: 10.3389/fpls.2021.704958

FIGURE 2.

FIGURE 2

Retarded growth of reproductive organs and degenerated pollen production and germination found in AIF2ox plants. (A) Flowers of Col-0, AIF2ox, aif2-1 and aif2-1/aif4-1 plants and the lengths of pistils, stamens, and the ratio of stamen to pistil. Number of open flowers examined >30. (B) Measurement of pollen numbers counted under a bright microscope without pollen staining. Number of open flowers examined >20. (C) In vitro pollen germination analysis. The efficiency of germination is represented by the ratio of germinated pollens after 6 h incubation in the germination medium. Number of pollens examined >3,000 taken from 20–25 flowers. (D) Frequency and size of the Alexander solution -stained viable pollens (normal) and non-stained aborted pollens. Number of pollens examined >2,000 taken from 15–20 open flowers (E) Defective pollen development in AIF2ox flowers. Pollens in different floral stages were stained with DAPI to reveal pollen developmental stages and pollens with intact nuclei (normal), without (aborted) or abnormal nuclei having defects in mitosis and appearance (aberrant). Number of pollens examined >1,000 taken from flowers at stage 4–5 or stage 11-12. (F,G) In vivo pollen tube growth assay. Arrowhead indicates the end of aniline blue-stained pollen tubes at 6 or 12 h after hand-pollinated self-pollination in the same flowers (F) and lengths of pollen tubes at different times were measured after hand-pollination (G). n > 15 for each time point was used for analysis. Statistical difference from either the Col-0 control or bracketed samples is indicated by two asterisks (**) at P < 0.01.