Dirigent gene editing of gossypol enantiomers for toxicity-depleted cotton seeds

This study identified two dirigent proteins responsible for the biosynthesis of the two enantiomers of gossypol in cotton. Editing one of the dirigent proteins selectively removed the toxic (-)-gossypol in cotton seeds without affecting pest resistance. Axial chirality of biaryls can generate varied bioactivities. Gossypol is a binaphthyl compound made by cotton plants. Of its two axially chiral isomers, (-)-gossypol is the bioactive form in mammals and has antispermatogenic activity, and its accumulation in cotton seeds poses health concerns. Here we identified two extracellular dirigent proteins (DIRs) from Gossypium hirsutum, GhDIR5 and GhDIR6, which impart the hemigossypol oxidative coupling into (-)- and (+)-gossypol, respectively. To reduce cotton seed toxicity, we disrupted GhDIR5 by genome editing, which eliminated (-)-gossypol but had no effects on other phytoalexins, including (+)-gossypol, that provide pest resistance. Reciprocal mutagenesis identified three residues responsible for enantioselectivity. The (-)-gossypol-forming DIRs emerged later than their enantiocomplementary counterparts, from tandem gene duplications that occurred shortly after the cotton genus diverged. Our study offers insight into how plants control enantiomeric ratios and how to selectively modify the chemical spectra of cotton plants and thereby improve crop quality.

Automated summary

This study identified two dirigent proteins responsible for the biosynthesis of the two enantiomers of gossypol in cotton. Editing one of the dirigent proteins selectively removed the toxic (-)-gossypol in cotton seeds without affecting pest resistance. Our study offers insight into how plants control enantiomeric ratios and how to selectively modify the chemical spectra of cotton plants and thereby improve crop quality.