Abstract

Lignin is a polymer in secondary cell walls of plants that is known to have negative impacts on forage digestibility, pulping efficiency, and sugar release from cellulosic biomass. While targeted modifications of different lignin biosynthetic enzymes have permitted the generation of transgenic plants with desirable traits, such as improved digestibility or reduced recalcitrance to saccharification, some of the engineered plants exhibit monomer compositions that are clearly at odds with the expected outcomes when the biosynthetic pathway is perturbed. In Medicago, such discrepancies were partly reconciled by the recent finding that certain biosynthetic enzymes may be spatially organized into two independent channels for the synthesis of guaiacyl (G) and syringyl (S) lignin monomers. Nevertheless, the mechanistic details, as well as the biological function of these interactions, remain unclear. To decipher the working principles of this and similar control mechanisms, we propose and employ here a novel computational approach that permits an expedient and exhaustive assessment of hundreds of minimal designs that could arise in vivo. Interestingly, this comparative analysis not only helps distinguish two most parsimonious mechanisms of crosstalk between the two channels by formulating a targeted and readily testable hypothesis, but also suggests that the G lignin-specific channel is more important for proper functioning than the S lignin-specific channel. While the proposed strategy of analysis in this article is tightly focused on lignin synthesis, it is likely to be of similar utility in extracting unbiased information in a variety of situations, where the spatial organization of molecular components is critical for coordinating the flow of cellular information, and where initially various control designs seem equally valid.

Highlights

  • Lignin is a phenolic heteropolymer in the secondary cell walls of vascular plants

  • The secondary cell walls of vascular plants contain several polysaccharides, such as cellulose and hemicellulose. Extraction of these polymers from lignocellulosic biomass for biofuel production has attracted extensive interest partly because exploitation of this source of fermentable sugars could minimize the competition for food, which has been criticized in the case of corn- or sugarcane-based biofuel production

  • We analyze metabolic channels in the biosynthetic pathway of lignin, a complex polymer that stiffens and fortifies secondary cell walls within woody tissues. This system is well suited because the present analysis can be based on a computational, experimentally validated model demonstrating that several enzymes are spatially associated into channels specific for the production of two lignin monomers

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Summary

Introduction

Lignin is a phenolic heteropolymer in the secondary cell walls of vascular plants. It is derived mainly from three hydroxycinnamyl alcohol monomers, namely p-coumaryl, coniferyl, and sinapyl alcohols, which, when incorporated into the lignin polymer, give rise to p-hydroxyphenyl (H), guaiacyl (G), and syringyl (S) subunits, respectively. While impressive advances have been made toward the reduction of biomass recalcitrance [3,4], it was shown that the amount of fermentable sugars released through chemical and enzymatic treatments is inversely proportional to that of lignin present in biomass, and that some transgenic plants with reduced lignin content yield up to twice as much sugar from their stems as wild-type plants [5] These observations suggest that lignin biosynthesis may be targeted for generating engineered crops with reduced recalcitrance. The challenge of this task derives from the fact that a rational design of less recalcitrant varieties would require a thorough, multi-level understanding of the lignin biosynthesis in wild- type plants, which we do not yet have. Such an understanding would include a grasp of the system of interactions between the enzymeencoding genes, proteins and metabolites involved in the biosynthesis of lignin, as well as details of the regulation of this multi-tiered system

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