Two ancestral APETALA3 homologs from the basal angiosperm Magnolia wufengensis (Magnoliaceae) can affect flower development of Arabidopsis

Abstract

APETALA3 (AP3) homologs are involved in specifying petal and stamen identities in core eudicot model organisms. In order to investigate the functional conservation of AP3 homologs between core eudicots and basal angiosperm, we isolated and identified two AP3 homologs from Magnolia wufengensis, a woody basal angiosperm belonging to the family Magnoliaceae. Sequence and phylogenetic analyses revealed that both genes are clade members of the paleoAP3 lineage. Moreover, a highly conserved motif of paleoAP3 is found in the C-terminal regions of MAwuAP3_1/2 proteins, but the PI-derived motif, usually present in AP3/DEF-like lineage members, is missing. Semi-quantitative and real time PCR analyses showed that the expression of MAwuAP3_1/2 was restricted to tepals and stamens. However, the MAwuAP3_1 expression was maintained at a high level during the rapid increased in size of tepals and stamens, while MAwuAP3_2 mRNA was only detected at the early stage of tepal and stamen development. Furthermore, the expression of MAwuAP3_1/2 in transgenic Arabidopsis causes phenotypic changes which partly resemble those caused by ectopic expressions of the endogenous AP3 gene. Moreover, the 35S::MAwuAP3_1/2 transgenic Arabidopsis can be used partially to rescue the loss-of-function ap3 mutant (ap3-3) of Arabidopsis. These findings call for a more comprehensive understanding of the B-functional evolution from basal angiosperm to core eudicot clades.

Keywords: AP3; AP3/DEF; APETALA3; APETALA3/DEFICIENS; B-class MADS box gene; BLAST; Basic Local Alignment Search Tool; DNA complementary to RNA; Dnase; Flower development; Magnolia wufengensis; ORF; PCR; PI; PISTILLATA; Polymerase chain reaction; QPCR; RACE; RNase; RT-PCR; UTR; base pair(s); bp; cDNA; dCAPS; deoxyribonuclease; derived cleaved amplified polymorphic sequence; open reading frame; quantitative polymerase chain reaction; rapid amplification of cDNA ends; reverse transcription polymerase chain reaction; ribonuclease; untranslated region(s).