Variance in Calvin-Benson cycle intermediate levels between closely-related species in the tomato clade

Abstract Published studies have reported species-variance between profiles of Calvin-Benson cycle (CBC) intermediates, not only between C4 species and C3 species, but also within C3 species (Arrivault et al., 2019, Borghi et al. 2019). It was proposed that this variance reflects lineage-dependent changes in the balance between different reactions, or poising, of the CBC. These earlier studies investigated phylogenetically-unrelated C3 species. In the current study, CBC intermediates were profiled in five closely-related species from Solanum sect. lycopersicon subsect. Lycopersicum. The levels of individual CBC intermediates showed many significant differences. In a principal component analysis, whilst three species (Solanum lycopersicum, Solanum cheesmaniae, Solanum neorickii) overlapped, Solanum pimpinellifolium and especially Solanum pennellii grouped separately, and were at opposing ends of the distribution. When combined with published data, whilst the separation between Solanum species was retained, they formed a group that was separated from five other C3 species, as well as two C4 species. It is discussed that the observed variation in CBC metabolites profiles within Solanum, together with their separation from other C3 species, supports the idea that CBC evolution is shaped both by phylogenetic relatedness and lineage-specific adaptation. Highlight Variance of intermediate levels points to poising of the Calvin-Benson cycle varying between closely-related species in the tomato clade Solanum sect. lycopersicon subsect. Lycopersicum Competing Interest Statement The authors have declared no competing interest. Footnotes Author emails: Vittoria Clapero, Clapero{at}mpimp-golm.mpg.de; vittoria.clapero{at}gmail.com; Stéphanie Arrivault, stephanie.arrivault{at}gmail.com; Mark Stitt, mstitt{at}mpimp-golm.mpg.de. Abbreviations - 2PG - 2-phosphoglycolate - 3PGA - 3-phosphoglycerate - CBC - Calvin-Benson cycle - CO2 - carbon dioxide - DHAP - dihydroxyacetone phosphate - F6P - fructose 6-phosphate - FBP - fructose 1,6-bisphosphate - FBPase - fructose-1,6-bisphosphatase - LC-MS/MS - liquid chromatography coupled with tandem mass spectrometry - Liquid N2 - liquid nitrogen - O2 - molecular oxygen - PC - principal component - PRK - phosphoribulokinase - R5P - ribose 5-phosphate - Ru5P - ribulose 5-phosphate - Rubisco - ribulose-bisphosphate carboxylase-oxygenase - RuBP - ribulose 1,5-bisphosphate - S7P - sedoheptulose 7-phosphate - SBP - sedoheptulose 1,7-bisphosphate - SBPase - sedoheptulose-1,7-bisphosphatase - Xu5P - xylulose 5-phosphate