Abstract
Introduction:
Plant-fungal interactions stimulate endophytic fungi to produce a plethora of metabolites that enhance plant growth and improve stress resistance. Opines, naturally occurring compounds formed through the condensation of amino acids with α-keto acids or sugars, have diverse biological functions and are mainly present in bacteria. Interestingly, investigations have revealed the presence of opine synthases (OSases) in fungal species as well, and their functions are yet to be studied.
Objectives:
The objective of this study is to investigate the occurrence of OSases in fungal species, identify their products, and characterize the potential biological activity of the metabolites.
Methods:
We identified a putative class of OSases in fungi through sequence similarity network (SSN) analysis. The function of these enzymes was elucidated using methods including protein heterologous expression, in vitro biochemical characterization, in vivo gene knock-out, as well as product isolation and identification. Additionally, we conducted plant activity testing on the secondary metabolites through foliar spraying and performed transcriptomic analysis to uncover their functions.
Results:
A quarter of the PF18631 family members, which contain the C-terminal helical bundle domain of cucumopine synthase, are derived from endophytic fungi. Some of these enzymes catalyze the synthesis of tryptopine A (1-acetyl-3-carboxy-β-carboline) by condensing L-tryptophan and methylglyoxal. The tryptopine A can act as a growth regulator, promoting plant growth and transcriptionally reprogramming photosynthesis-related pathways, while enhancing the rate of plant photosynthesis by 25%.
Conclusion:
The findings of this study suggest that tryptopine A plays a crucial role as a signaling molecule in the establishment and maintenance of mutualistic associations between endophytic fungi and host plants, thereby broaden our comprehension of fungal-plant symbiosis.
Keywords:
Endophytic fungi; Opine synthase; Plant growth regulator; Secondary metabolite.