Reduced γ-glutamyl hydrolase activity likely contributes to high folate levels in Periyakulam-1 tomato

Abstract

Tomato cultivars show wide variation in nutraceutical folate in ripe fruits, yet the loci regulating folate levels in fruits remain unexplored. To decipher regulatory points, we compared two contrasting tomato cultivars: Periyakulam-1 (PKM-1) with high folate and Arka Vikas (AV) with low folate. The progression of ripening in PKM-1 was nearly similar to AV but had substantially lower ethylene emission. In parallel, the levels of phytohormones salicylic acid, ABA, and jasmonic acid were substantially lower than AV. The fruits of PKM-1 were metabolically distinct from AV, with upregulation of several amino acids. Consistent with higher ◦Brix, the red ripe fruits also showed upregulation of sugars and sugar-derived metabolites. In parallel with higher folate, PKM-1 fruits also had higher carotenoid levels, especially lycopene and β-carotene. The proteome analysis showed upregulation of carotenoid sequestration and folate metabolism-related proteins in PKM-1. The deglutamylation pathway mediated by γ-glutamyl hydrolase (GGH) was substantially reduced in PKM-1 at the red-ripe stage. The red-ripe fruits had reduced transcript levels of GGHs and lower GGH activity than AV. Conversely, the percent polyglutamylation of folate was much higher in PKM-1. Our analysis indicates the regulation of GGH activity as a potential target to elevate folate levels in tomato fruits.

Acknowledgements

This work was supported by the Department of Biotechnology (DBT), India grants, BT/PR11671/PBD/16/828/2008, BT/PR/7002/PBD/16/1009/2012, and BT/COE/34/SP15209/2015 to RS and YS. We also acknowledge fellowships by CSIR to KT and UGC to AS and HVK.

Author contributions

KT, YS, and RS designed the research and wrote the manuscript; KT did most experiments, AS did proteome data analysis, and HVK did proteome profiling. PG analyzed the AV and PKM-1 genome sequence.

Conflict of Interest

The authors declare no conflict of interest.

Data Availability

The data consisting of metabolite levels and proteome is included as supplemental material. The proteome data are uploaded to the PRIDE repository, which can be accessed using the following details: Project Name: PKM-1 proteome; Project accession: PXD027940.

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