AtENO2 functions in the development of male gametophytes in Arabidopsis thaliana

Pollen fertility is an important factor affecting the seed setting rate and seed yield of plants. The Arabidopsis thaliana enolase gene ENO2 (AtENO2) can affect the pollen morphology, germination, and pollen tube growth. AtENO2 encodes two proteins AtENO2 and AtMBP-1. To examine the effect of AtENO2 protein on pollen development, the 2nd ATG of the AtENO2 coding sequence for AtMBP-1 was mutated by site-directed mutagenesis, and transgenic plants expressing only AtENO2 but not AtMBP-1 were obtained. Phenotypic analysis indicated that AtENO2 was essential in the pollen development. The mechanisms of AtENO2 on pollen development were analyzed. AtENO2 can affect development of the pollen intine, and the mechanism may be that AtENO2 regulated the methyl esterification of pectin in pollen intine through ARF3 and AtPMEI-pi. The -734 ∼ -573 sequence of AtENO2 promoter is the main transcriptional regulatory region of AtENO2 affecting pollen development. The functional cis-acting element may be GTGANTG10(GTGA), and the trans-acting factors may be KAN, AS2 and ARF3/ETT. Moreover, the deletion of AtENO2 can cause significant difference in the expression of multiple genes related to pollen exine development. These results are useful for further studying the function of AtENO2 and exploring the mechanism of plant pollen development.