A dolabralexin-deficient mutant provides insight into specialized diterpenoid metabolism in maize

Two major groups of specialized metabolites in maize (Zea mays), termed kauralexins and dolabralexins, serve as known or predicted diterpenoid defenses against pathogens, herbivores, and other environmental stressors. To consider the physiological roles of the recently discovered dolabralexin pathway, we examined dolabralexin structural diversity, tissue-specificity, and stress-elicited production in a defined biosynthetic pathway mutant. Metabolomics analyses support a larger number of dolabralexin pathway products than previously known. We identified dolabradienol as a previously undetected pathway metabolite and characterized its enzymatic production. Transcript and metabolite profiling showed that dolabralexin biosynthesis and accumulation predominantly occur in primary roots and show quantitative variation across genetically diverse inbred lines. Generation and analysis of CRISPR-Cas9-derived loss-of-function Kaurene Synthase-Like 4 (Zmksl4) mutants demonstrated dolabralexin production deficiency, thus supporting ZmKSL4 as the diterpene synthase responsible for the conversion of geranylgeranyl pyrophosphate precursors into dolabradiene and downstream pathway products. Zmksl4 mutants further display altered root-to-shoot ratios and root architecture in response to water deficit. Collectively, these results demonstrate dolabralexin biosynthesis via ZmKSL4 as a committed pathway node biochemically separating kauralexin and dolabralexin metabolism, and suggest an interactive role of maize dolabralexins in plant vigor during abiotic stress.

Automated summary

In maize, two major groups of specialized metabolites, kauralexins and dolabralexins, are involved in defending against pathogens, herbivores, and other stressors. This study examined the dolabralexin pathway and found new metabolites and characterized their production. Genetic analysis showed that dolabralexin biosynthesis occurs mainly in primary roots and varies across different maize lines. Loss-of-function mutants for the diterpene synthase gene ZmKSL4 demonstrated deficient dolabralexin production and exhibited altered root architecture in response to water deficit. These findings suggest that maize dolabralexins play a role in plant vigor during abiotic stress.