Loss of GCS1/HAP2 does not affect the ovule-targeting behavior of pollen tubes.

KEY MESSAGE : hap2-1 pollen tube ovule targeting. Upon pollination, a pollen grain germinates to produce a pollen tube, which grows through the style to deliver two immobile sperm cells to the female gametophyte. Double fertilization is completed after the pollen tube enters an ovule. GENERATIVE CELL SPECIFIC 1 (GCS1)/HAPLESS 2 (HAP2) contributes to the fusion of gametes at fertilization and has been suggested to affect pollen tube guidance. However, there is controversy over the role of GCS1/HAP2 in pollen tube guidance because of conflicting results from different studies. To characterize the effects of the gcs1/hap2 mutation on pollen tube behavior, we analyzed the Arabidopsis thaliana hap2-1/HAP2 mutant, which carries a gcs1/hap2 mutation in the quartet background. The quartet mutant produces tetrads consisting of four pollen grains that remain adherent after the pollen mother cell has completed meiosis. Thus, a hap2-1/HAP2 tetrad contains hap2-1 and HAP2 pollen grains in a 2:2 ratio. Moreover, the hap2-1 locus is linked to the β-glucuronidase (GUS) gene. An excess pollination experiment with hap2-1/HAP2 tetrads revealed that the hap2-1 pollen tube targets ovules normally. Additionally, the results of restricted pollination and aniline blue staining indicated that there are no significant differences between the ovule-targeting frequencies of pollen tubes from hap2-1/HAP2 and HAP2/HAP2 tetrads.