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Respiratory burst oxidase homologs (Rbohs) play a predominant role in reactive oxygen species (ROS) production, which is crucial in plant growth, differentiation, as well as their responses to biotic and abiotic stresses. To date, however, there is little knowledge about the function of cotton  Rboh  genes. Here, we identified a total of 87  Rbohs  from five sequenced  Gossypium  species (the diploids  Gossypium arboreum ,  Gossypium raimondii , and  Gossypium australe , and the allotetraploids  Gossypium hirsutum  and  Gossypium barbadense ) via BLAST searching their genomes. Phylogenetic analysis of the putative 87 cotton  Rbohs  revealed that they were divided into seven clades. All members within the same clade are generally similar to each other in terms of gene structure and conserved domain arrangement. In  G. barbadense , the expression levels of  GbRbohs  in the CladeD were induced in response to a fungal pathogen and to hormones (i.e., jasmonic acid and abscisic acid), based upon which the main functional member in CladeD was discerned to be  GbRboh5/18.  Further functional and physiological analyses showed that the knock-down of  GbRboh5/18  expression attenuates plant resistance to  Verticillium dahliae  infection. Combined with the molecular and biochemical analyses, we found less ROS accumulation in  GbRboh5/18-VIGS  plants than in control plants after  V. dahliae  infection. Overexpression of  GbRboh5/18  in  G. barbadense  resulted in more ROS accumulation than in control plants. These results suggest that  GbRboh5/18  enhances the cotton plants’ resistance against  V. dahliae  by elevating the levels of ROS accumulation. By integrating phylogenetic, molecular, and biochemical approaches, this comprehensive study provides a detailed overview of the number, phylogeny, and evolution of the  Rboh  gene family from five sequenced  Gossypium  species, as well as elucidating the function of  GbRboh5/18  for plant resistance against  V. dahliae . This study sheds fresh light on the molecular evolutionary properties and function of  Rboh  genes in cotton, and provides a reference for improving cotton’s responses to the pathogen  V. dahliae .

frontiers in genetics

Comprehensive Analysis of Respiratory Burst Oxidase Homologs (Rboh) Gene Family and Function of GbRboh5/18 on Verticillium Wilt Resistance in Gossypium barbadense