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Glycolate oxidase (GOX)-dependent production of H 2 O 2  in response to pathogens and its function in disease resistance are still poorly understood. In this study, we performed genome-wide identification of  GOX  gene family in  Nicotiana benthamiana  and analyzed their function in various types of disease resistance. Sixteen  GOX  genes were identified in  N .  benthamiana  genome. They consisted of GOX and HAOX groups. All but two NbGOX proteins contained an alpha_hydroxyacid_oxid_FMN domain with extra 43–52 amino acids compared to that of FMN-dependent alpha-hydroxyacid oxidizing enzymes (NCBI-CDD cd02809). Silencing of three NbGOX family genes  NbHAOX8 ,  NbGOX1  and  NbGOX4  differently affected resistance to various pathogens including  Tobacco rattle virus ,  Xanthomonas oryzae  pv.  oryzae  ( Xoo ) and  Sclerotinia sclerotiorum . Effect of these genes on resistance to  Xoo  is well correlated with that on  Xoo –responsive H 2 O 2  accumulation. Additionally, silencing of these genes enhanced PAMP-triggered immunity as shown by increased flg22-elicited H 2 O 2  accumulation in  NbGOX -silenced plants. These NbGOX family genes were distinguishable in altering expression of defense genes. Analysis of mutual effect on gene expression indicated that  NbGOX4  might function through repressing  NbHAOX8  and  NbGOX1 . Collectively, our results reveal the important roles and functional complexity of  GOX  genes in disease resistance in  N .  benthamiana .

Glycolate oxidase gene family in Nicotiana benthamiana: genome-wide identification and functional analyses in disease resistance