RNA-Seq analysis reveals the distinctive adaxial-abaxial polarity in the asymmetric one-theca stamen of Canna indica.

Proper establishment of adaxial-abaxial polarity is essential for the development of lateral organs, while former researches were mostly focused on the polarity regulation in leaves, and little is known in stamens, especially in the asymmetric ones. Canna indica (Zingiberales: Cannaceae) is a widely cultivated ornamental plant and the representative species to study the evolutionary development of Zingiberales. The androecium of Canna indica comprises 3-4 petaloid staminodes and a fertile stamen (FS), which consists of a one-theca anther and a petaloid appendage. The partially petaloid stamen is considered as an intermediate state organ from a two-thecae stamen to a completely petaloid staminode. Using RNA-Seq, we quantified the expressions of the transcripts in anther and petaloid appendage, and detected 64,430 and 57,041 unigenes in these two organs, respectively. 4574 unigenes were down-regulated, and 3525 were up-regulated in petaloid appendage compared with those in anther. GO enrichment analysis indicated that the function of cytokinin is more related to cell differentiation in anther, while auxin is more to cell division in petaloid appendage. B- and C-class floral homeotic genes were expressed in these two androecium parts. Most of the class III HD-ZIP family members, which specify adaxial identity, were expressed lower in petaloid appendage than in anther; while KANADIs and YABBYs, which promote abaxial identity, exhibited opposite expression patterns. In situ hybridization showed that the adaxial marker gene was mainly expressed in the region between the two protrusions of the anther, while the abaxial marker was mainly expressed in petaloid appendage. We hypothesize that the adaxial-abaxial polarity participates in the distinctive anther-petaloid appendage patterning within the asymmetric FS of Canna indica.