Fig. 2.
Loss of CG and non‐CG DNA methylation impairs pollen development and fertilisation in Arabidopsis. (a–e) 4′,6‐Diamidino‐2‐phenylindole (DAPI) staining of mature pollen grains from Col‐0, met1‐11+/− , met1‐11, ddcc and ddcc met1+/− mutant plants. (e1–e3) White arrows indicate pollen grains that are abnormally binucleated (e1), non‐nucleated (e2), and mononucleated (e3) in ddcc met1+/− . To confirm pollen identity for non‐nucleated (e2) and mononucleated (e3) pollen in ddcc met1+/− , images under bright field are showed. Bars, 100 μm. (f) Percentages of phenotypically abnormal pollen grains in Col‐0, met1‐11+/− , met1‐11, ddcc and ddcc met1+/− mutant plants (n > 500, for each genotype). (g–i) Aniline blue staining showing pollen tube access to each ovule at 2 d after pollination in Col‐0 ♀× Col‐0 ♂ (g) and Col‐0 ♀× ddcc met1+/− ♂ (h) siliques. Ovules that have normal pollen tube acceptance but cannot expand (failed ovules, fo) are frequently found in the Col‐0 ♀×ddcc met1+/− ♂ siliques (i). White arrows indicate normal pollen tube acceptance in a failed ovule and a developing seed (ds). Bars, 100 μm. (j) Percentages of developing seeds (ds) and failed ovules (fo) in Col‐0 ♀× Col‐0 ♂, Col‐0 ♀×met1‐11 ♂, Col‐0 ♀×ddcc ♂, and Col‐0 ♀× ddcc met1+/ − ♂ siliques (n > 200, for each genotype).