Protein phosphatase 2A B'α and B'β promote pollen wall construction partially through BRASSINAZOLE-RESISTANT 1-activated cysteine protease gene CEP1 in Arabidopsis.

A well-constructed pollen wall is essential for pollen fertility, which relies on the contribution of the tapetum. Our results demonstrated an essential role of the tapetum-expressed protein phosphatase 2A (PP2A) B'α and B'β in pollen wall formation. The b'aβ double mutant pollen grain harboured sticky remnants and tectum breakages, resulting in failed release. B'α and B'β functioned partially through dephosphorylating and activating BRASSINAZOLE-RESISTANT 1 (BZR1). The bzr1 bes1 double and higher-order mutants of this BZR1/BES1 family displayed similar defects in the pollen wall, while bzr1-1D, having an active form of the BRZ1 protein, exhibited fertile pollen grains in a B'α and B'β dependent manner. Correspondingly, the level of phospho-BZR1 was increased and dephospho-BZR1 was decreased in b'aβ and bzr1-1D/b'aβ at anther stages 8-9 as compared with Col-0 and bzr1-1D, respectively. A cysteine protease gene CEP1 was identified as a BZR1 target, whose transcriptional activation necessitates brassinosteroid (BR)-responsive elements in the promoter region and the BZR1 DNA binding domain. The mRNA level of CEP1 at stages 8-9 was extremely low in bzr1 and bzr1 bes1, but higher in Col-0 and bzr1-1D depending on B'α and B'β. Furthermore, cep1 mutants displayed similar defects in the pollen wall. In brief, this study uncovered a PP2A-BZR1-CEP1 regulatory module, providing a new insight into pollen maturation mechanisms.