Neofunctionalization of a Polyploidization-Activated Cotton Long intergenic non-coding RNA DAN1 During Drought Stress Regulation.

The genomic shock of whole-genome duplication and hybridization introduces great variation into transcriptomes, for both coding and non-coding genes. An altered transcriptome provides a molecular basis for improving adaptation during the evolution of new species. The allotetraploid cotton, together with the putative diploid ancestor species compose a fine model for study the rapid gene neofunctionalization over the genome shock. Here we report on Drought-Associated Non-coding gene 1 (DAN1), a long intergenic non-coding RNA (lincRNA) that arose from the cotton progenitor A-diploid genome after hybridization and whole-genome duplication events during cotton evolution. DAN1 in allotetraploid upland cotton (Gossypium hirsutum, Gh) is a drought-responsive lincRNA predominantly expressed in nucleoplasm. A chromatin isolation by RNA purification (ChIRP) profiling and electrophoretic mobility shift assay (EMSA) analysis demonstrated that GhDAN1 RNA can bind with DNA fragments containing AAAG motifs, similar to DNA-binding one zinc finger (Dof) transcription factor binding sequences. The suppression of GhDAN1 mainly regulates genes with AAAG motifs in auxin-response pathways, which are associated with drought stress regulation. As a result, GhDAN1-silenced plants exhibit improved tolerance to drought stress. This phenotype resembles the drought-tolerant phenotype of the A-diploid cotton ancestor species, which has an undetectable expression of DAN1. The role of DAN1 in cotton evolution and drought tolerance regulation suggests that the genomic shock of interspecific hybridization and whole-genome duplication (WGD) stimulated neofunctionalization of non-coding genes during the natural evolutionary process.